Acomys cahirinus). Here, we examine how hair melanin content of spiny mice living in the opposing slopes of EC evolves toward blending with their respective background habitat.Coat coloration in mammals is an explicit adaptation through natural selection. Camouflaging with the environment is the foremost evolutionary drive in explaining overall coloration. Decades of enquiries on this topic have been limited to repetitive coat color measurements to correlate the morphs with background/habitat blending. This led to an overwhelming endorsement of concealing coloration as a local phenotypic adaptation in animals, primarily rodents to evade predators. However, most such studies overlooked how rodents actually achieve such cryptic coloration. Cryptic coloration could be attained only through optimization between the yellow- to brown-colored “pheomelanin” and gray to black-colored “eumelanin” in the hairs. However, no study has explored this conjecture yet. “Evolution Canyon” (EC) in Israel is a natural microscale laboratory where the relationship between organism and environment can be explored. EC is comprised of an “African” slope (AS), which exhibits a yellow-brownish background habitat, and a “European” slope (ES), exhibiting a dark grayish habitat; both slopes harbor spiny mice (A. cahirinus approximates the background color of the slope on which they dwell. Pheomelanin is slightly (insignificantly) higher in individuals found on the AS to match the brownish background, whereas individuals of the ES had significantly greater eumelanin content to mimic the dark grayish background. This is further substantiated by a significantly higher eumelanin and pheomelanin ratio on the ES than on the AS.We measured hair-melanin (both eumelanin and pheomelanin) contents of 30 spiny mice from the EC using high-performance liquid chromatography (HPLC) that detects specific degradation products of eumelanin and pheomelanin. The melanin pattern of It appears that rodents adaptively modulate eumelanin and pheomelanin contents to achieve cryptic coloration in contrasting habitats even at a microscale. Peromyscus crinitus) Peromyscus maniculatus;) Peromyscus polionotus) Chaetodipus intermedius) The study of phenotypic responses to the environment embraces a long, enlightened history per se has been hypothesized to serve three main adaptive functions: intraspecific communication, concealment (crypsis), and regulation of physiological processes such as thermoregulation, UV protection, etc. Animals attain concealment when overall body color resembles their background in which they live There is ample evidence documenting what proportion of functional colors and patterns represent background color-matching Acomys cahirinus), which exhibit slope-specific adaptations to various selection pressures (http://evolution.haifa.ac.il].The “Evolution Canyon” model A. cahirinus) to mimic differently colored habitats between the slopes . Individuals of A. cahirinus on the opposing slopes exhibit visible differences in their overall coat color: AS individuals tend to be lighter and ES individuals tend to be darker  where the demand is to mimic yellow-brown backgrounds and higher eumelanin content on the ES (than on the AS) where the demand is to mimic dark gray backgrounds to increase survival fitness by reducing the potential to be detected by predators.The “African” slope is characterized by light terra rossa soil with a stretch of yellow grass cover displaying a yellow-brown colored milieu ; by conte darker . Here weAcomys cahirinus exhibit differences between the two (interslope) contrasting habitats. Mean eumelanin concentration in the hairs from ES/NFS individuals  was significantly  = 2.063, P = 0.024; (28) = 1.145, P = 0.137; (28) = 1.759, P = 0.044;  = 3.68, P = 0.0129;  = 2.70, P = 0.045, A. cahirinus captured from different stations of EC, while eumelanin  = 2.02, P = 0.111; (12) = 0.752, P = 0.467) and pheomelanin  = 0.0448, P = 0.965) did not vary significantly on the AS. This was also true in the ES, wherein eumelanin  = 1.462, P = 0.166) and pheomelanin  = 0.552, P = 0.590) contents of males and females did not vary.Hair melanin contents of A. cahirinus) approximates the background (habitat) color. Pheomelanin is insignificantly higher in individuals from the “African” slope to harmonize with the yellow to brownish background that is characterized by light terra rossa soil covered with a stretch of yellow-brownish grass across the slope ” and “European Slope (ES)” vary in their melanin contents? The most likely explanation embraces a series of biochemical facts. Indeed, the pigmentation of hair in mammals mainly depends on the quantity, quality, and distribution of melanin. Natural melanin pigments consist of both insoluble brown to black eumelanin and alkali-soluble pheomelanin in varying ratios and are thus often considered “mixed melanin” One possible way by which the spiny mice modulate the amounts of eumelanin and pheomelanin, is adjusting the size/length of the agouti bands in hairs. Substantially, our results provide some clues that the concentration of pheomelanin fluctuated significantly between stations compared to that of eumelanin, which only led to significant variation in the eumelanin/pheomelanin ratio across stations. This indirectly signifies that the mice probably adjust the size of the agouti bands, or, in other words, they modulate the distribution and accumulation of pheomelanin in hairs to achieve background (habitat) matching. Presumably, the higher the pheomelanin contents derive from the greater the size/length of the agouti band Acomys cahirinus colonized Israel in the Upper Pleistocene, approximately 30–20,000 years ago Mc1r gene) and its antagonist, the agouti signaling protein (product of Agouti gene), are the two candidate genes responsible for most of the inter-specific as well as intra-specific differences in pigmentation melanocortin-1 receptor (Mc1r), have been shown to be solely responsible in the majority of molecular underpinning of phenotypic variation. However, a recent study on melanism in Peromyscus showed independent mutations in agouti itself demonstrating that mutations in genes other than Mc1r can still produce changes in melanin At the molecular level, melanocortin-1 receptor  A previous study in our lab examined genetic allozyme and RAPD diversities to understand the ecological-genetic patterns in the spiny mouse The phenomenon of crypsis through their coat/pelage coloration in rodents is very well established for over a century A. cahirinus provides evidence for background matching and/or endorses Gloger's rule. Gloger's rule ascertains that darker individuals tend to reside in more humid regions and paler individuals in drier areas, possibly optimizing thermoregulation A. cahirinus may respond more towards background matching than microclimatic factors, we cannot rule out the possibility of both causes. Remarkably, regionally in Israel A. cahirinus is darker in the Galilee and lighter in the Negev desert and this is partly attributed to thermoregulatory processes Mus musculus) reveals that dark-colored coats of mice were observed in more humid and closed habitats with darker colored backgrounds, while pale coats were found in drier, more open habitats with lighter background color Hair color is the most obvious and diverse characteristic among pigmentation phenotypes. The quality of habitats can influence animal color A. cahirinus, we propose the hypothesis that through natural selection rodents modulate or switch  eumelanin and pheomelanin contents in their coat/pelage to achieve background matching in contrastingly colored habitats/backgrounds. Large-scale and critical studies that focus on cross-species comparison of hair melanin evolution towards background matching are needed to test our hypotheses. The role of Mc1r and agouti genes in effecting such pigment switching or modulation in spiny mice needs to be studied in detail. Furthermore, ecological transplant experiments and genetic crosses between mice of the opposite slopes of “Evolution Canyon” to assess the Mendelian genetics and levels of natural selection involved, are direct natural extensions of the current study.Based on our results with “Evolution Canyon” I, located at lower Nahal Oren , is a deeply erosive valley running from Mount Carmel westward into the Mediterranean Sea. The opposite slopes of the canyon have identical regional evolutionary history, geology, and soil type Ceratonia siliqua, Pistacia lentiscus, and savanna grasses . In contrast, the “European” north-facing slope (ES) is covered by an east Mediterranean maquis-forest (brushwood forest) of evergreen live oak of Quercus calliprinos, and Pistacia palaestinaThe “African” south-facing slope (AS) is covered by xeric, Mediterranean, savannoid formation of Acomys cahirinus) were trapped from the opposing slopes  of “Evolution Canyon” I, Nahal Oren, Carmel, Israel. A total of 30 mice were captured from the opposing slopes; 14 were caught from the AS including the upper, middle, and lower parts (stations AS1 (n = 5), AS2 (n = 3), and AS3 (n = 6), respectively), and 16 were caught from the ES (stations ES7 (n = 6), ES6 (n = 5), and ES5 (n = 5), respectively). Animals were housed in individual cages. They were kept under controlled conditions at 22–24°C and fed carrots and cucumbers. Animals used in this study were adults. The experiments were approved by the Ethics Committee of the University of Haifa. The mice were kept in the laboratory under 12∶12 LD (light/dark) cycle. Hair samples were carefully  excised with scissors from the rump region of animals over a 4×4 cm area, with uniform lengths . This procedure was consistent for all individuals to standardize sample collections for hair melanin analysis. Animals were treated humanely and care was taken while removing hairs to prevent injury or suffering.Individuals of spiny mice  and analyzed by HPLC with ultraviolet detection to determine eumelanin content A. cahirinus between AS/SFS and ES/NFS and between the sexes were compared using t-tests. Melanin contents are given as ng/mg. One-way analysis of variance (ANOVA), followed by the Waller-Duncan K-ratio t-test was used to compare the differences in hair-melanin contents of A. cahirinus across different stations in the opposing slopes of the EC. Data are shown as mean ± SE.The differences in hair melanin variables of


Osteosarcoma (OS) is the most common nonhematologic malignancy of bone in children and adults. Although dysregulation of tumor suppressor genes and oncogenes, such as Rb, p53, and the genes critical to cell cycle control, genetic stability, and apoptosis have been identified in OS, consensus genetic changes that lead to OS development are poorly understood. Disruption of the osteogenic differentiation pathway may be at least in part responsible for OS tumorigenesis. Current OS management involves chemotherapy and surgery. Peroxisome proliferator-activated receptor (PPAR) agonists and/or retinoids can inhibit OS proliferation and induce apoptosis and may inhibit OS growth by promoting osteoblastic terminal differentiation. Thus, safe and effective PPAR agonists and/or retinoid derivatives can be then used as adjuvant therapeutic drugs for OS therapy. Furthermore, these agents have the potential to be used as chemopreventive agents for the OS patients who undergo the resection of the primary bone tumors in order to prevent local recurrence and/or distal pulmonary metastasis. Osteosarcoma (OS) is the most common nonhematologic malignant tumor of bone in adults and children, with the peak incidence in early childhood , 2. It iIt has been shown that OS cells are similar to undifferentiated osteoblasts, and increasing evidence suggests that osteogenic differentiation defects may be responsible for OS tumorigenesis , 7–10. Oγ agonists has been shown in tumor cells derived from liposarcoma, colon cancer, breast cancer, leukemia, gastric cancer, nonsmall cell lung cancer, and prostate cancer . Tr. Trα1), γ : RXR interaction is seen in Familial Partial Lipodystrophy, an autosomal dominant condition associated with metabolic syndrome, characterized by dyslipidemia, abnormal adipose tissue distribution, and a number of metabolic abnormalities [γ that affect its ability to dimerize with RXR and bind coactivators.A unique association with PPAR and RXR is the direct repeat responsive elements (DR) and the PPAR response elements (PPRE) associated with the heterodimer. DR1 is a repetition of 2 core motifs consisting of AGGTCA spaced apart from one another by one nucleotide in the promoter of multiple target genes  63]. Th. Th63]. malities . This syγ is of particular interest because of its roles in adipogenesis, atherosclerosis, inflammation, proliferation, differentiation, and apoptosis [γ gene contains 3 promoters, producing 2 different proteins, PPARγ1 and PPARγ2, which likely contributes to this diverse range of functions [γ expression in vascular endothelium and smooth muscle cells leads to an inhibition of MMPs, downregulation of Angiotensin II type 1 receptor, and alteration of macrophage invasion [γ, leading to its activation in the inflammatory response cascade. Furthermore, the insulin sensitizing effects of PPARγ are demonstrated using the synthetic antidiabetic therapy thiazolinediones [γ functions in many tissues.PPARpoptosis . The PPAunctions . PPARγ einvasion –71. In iinvasion , 42, 72.nediones . Thus, tγ binds to fatty acids and their derivatives, such as linoleic acid and docosahaxaenoic acid (DHA) [γ, stimulating preadipocytes to differentiate and initiate the steps required in lipid storage. PPARγ effects are carried out through target genes, such as aP2, lipoprotein lipase (LPL), acyl-Coa synthetase (ACS), and CD36 [γ in fibroblasts initiates the adipogenic cascade, while PPARγ knockout mice are unable to form adipocytes or adipose tissue [γ leads to increased adipogenesis and obesity [γ activity results in lower body mass index [For adipogenesis, PPARid (DHA) , 43. Theand CD36 , 75. Ovee tissue , 49, 76. obesity . Anotherss index , 79.γ include LPL and fatty acid transporter FATP in stimulating fatty acid uptake, malic enzyme in NADPH synthesis for lipogenesis, phosphoenolpyruvate carboxykinase in gluconeogenesis, and ACS in FA esterification [γ promotes adipocyte differentiation of MSCs through various signaling pathways [γ activation [γ-associated adipogenic stimulation or osteogenic inhibition depends on the nature of the ligands [Adipose tissue targets of PPARfication , 80–82. pathways –85. MSCstivation –89. Howe ligands . Such liOsteogenesis and adipogenesis appear to originate from the same progenitor bone marrow mesenchymal stem cells (MSCs) –93. MSC γ can stimulate adipocyte differentiation of MSCs. PPAR+ blasts (progenitor cells) have been noted in hematopoeitic lineages, and PPARγ is able to induce terminal differentiation in monocytes and adipocytes [γ in fibroblastic cells initiates the adipogenic cascade, while PPARγ knockout mice were unable to form adipocytes or adipose tissue [γ leads to increased adipogenesis and obesity, while inactivating mutation results in a lower body mass [γ and almost all pathways involved in adipogenesis involve regulation of PPARγ [PPARipocytes , 113. Ove tissue , 49, 76.ody mass –79. To dof PPARγ .γ agonists were found to have a remarkable role regulating bone turnover [γ seems to shift the differentiation pathway away from osteoblastogenesis and towards osteoclastogenesis, this is not always the case. Giaginis et al. reviewed studies focusing on both the synthetic and natural PPARγ ligand effects on osteoblast and osteoclast formation, as well as apoptosis and overall bone formation. They found divergent results, as it appears there are other factors that contribute to bone turnover regulated by PPARγ. For example, since natural ligands are found in both the diet and the inflammatory cascade, perhaps these processes determine the final outcome of PPARγ-regulated bone turnover. Its effect in clinical studies poses a similar paradox, while some patients receiving synthetic PPARγ agonists for Diabetes Mellitus type II experienced bone loss, others were noted to have a decrease in bone resorption markers [Furthermore, in a review by Giaginis et al. PPARturnover . However markers .γ plays an intriguing role in both adipogenesis and osteogenesis. Earlier reports indicate that homozygous PPARγ deficient progenitor cells spontaneously differentiate into osteoblasts via increased osteoblastogenic factors in vitro, and heterozygous PPARγ deficiency results in increased in vivo bone formation [γ has been shown to be significantly upregulated by osteogenic BMPs [γ2 promotes the osteogenic BMP-induced osteogenesis and adipogenesis [γ2 expression leads to an inhibition of adipogenic differentiation as well as stimulation of osteogenic differentiation and osteoid matrix mineralization [PPARormation . Howeverormation , 117. PPnic BMPs , 118. Ovogenesis . Silencilization . Howeverγ and other members of its nuclear receptor superfamily. Regulation of the osteogenic promoter, osteocalcin, by glucocorticoids, vitamin D, and thyroid hormone, occurs through the same nuclear pathway as PPARγ [γ activation by fatty acids and their derivatives might lead to a slowing of osteoblast differentiation, which would explain the tendency to shift to adipogenesis. These findings are intriguing as recent studies have indicated that aging activates adipogenesis and suppresses osteogenesis, possibly through the increased availability of these fatty acids and a decrease in metabolic production of many nuclear hormones. These would shift the signaling towards PPARγ, which might explain part of the mechanism underlying osteoporosis [The regulation underlying these effects could be secondary to the nuclear competition between PPARas PPARγ –121. In oporosis , 123. γ ligands, the most common side effects observed have been headaches, gastrointestinal symptoms , and susceptibility to infections. Troglitazone has been withdrawn from the market secondary to its hepatic toxicity; however, this appears to be drug specific and not universal amongst PPAR agonists.In the treatment of Diabetes Mellitus (DM) type II with synthetic PPARγ agonists induce the cytochrome P450 isoform CYP3A4 in the liver, affecting the metabolism of many other drugs. These drugs have been shown to increase plasma volume, thus, leading to edema and a dilutional drop in hematocrit. While the mechanism has not been elicited, PPARγ agonists antagonize the vasoconstriction induced by hyperinsulinemia, by sensitizing cells to the effects of insulin. Therefore, they relax vascular smooth muscles and decrease peripheral blood pressure. While there is no mention of increased peripheral adipose tissue, the propensity of PPARγ to induce lipid storage might underlie the observed weight gain [In a review by Mudaliar and Henry about the clinical use of glitazones, side effects include edema, weight gain, and mild drops in hematocrit. Rarely, increases in liver enzymes are observed. These synthetic PPARght gain .γ agonists, there has been relatively little other evidence supporting any of these notions [Theoretically, a shift away from osteogenesis and towards adipogenesis might also promote osteoporosis and increase fracture risk. This has been demonstrated in a prospective study of over 80,000 patients being treated for DM type II . Further notions .INK4a, c-Myc, Fos-Jun, MDM2, CDK4, and cyclin D [β/BMP families, and IGF2 [The molecular pathogenesis underlying OS development is poorly understood. OS is associated with aberrations in p53 and Rb expression , 126–128and IGF2 –131. Theand IGF2 –134. Disruption of the osteogenic differentiation pathway from MSCs is thought at least in part to be responsible for OS tumorigenesis , 9, 10 . By previn vivo, which can be overcome by introducing key osteogenic regulator Runx2 [Therefore, investigation into the induction of terminal differentiation in cells with such differentiation defects has increased in recent years. A similar process has been reported in Ewing's Sarcoma, in which silencing the EWS/FLI-1 in Ewing's sarcoma cells leads to their recovery of MSC capability to differentiate into osteogenic lineages . Accordior Runx2 . These for Runx2 , 8. Theror Runx2 . DiffereIt appears that the differentiation status not only is responsible for the development of OS but also may predict its malignant potential. From the early 1970s when the idea of differentiation was first proposed, to more recently when differentiation agents are used for certain cancer phenotypes, it has been observed that this process is associated with many morphological changes in the respective cells. These changes leading to a well-differentiated cell include repression of responsiveness to growth factors, withdrawal from the cell cycle into a state of quiescence, and a decreased ability to re-initiate proliferation . For exaThe fundamental idea behind differentiation therapy for tumors is that by inducing terminal differentiation, the tumor cells lose their proliferative phenotypes. Differentiation causes cells to lose their proliferative potential and repress their responsiveness to growth factors, while at the same time possibly increasing their susceptibility to apoptosis, induction of tumor suppressors, repression of oncogenes, inhibition of angiogenesis, and induction of cytotoxic agents. As cells become more differentiated, these changes make them less aggressive and more responsive to other chemotherapeutic agents.Inhibition of tumor growth through differentiation therapy has been demonstrated in clinical cases of hematologic and breast tumors. Induction of terminal differentiation was first shown to be successful in treating AML with low-dosage araC . Recentlα is responsible for hepatocarcinogenic effects in rodents [β was identified as a downstream target of the APC/β-catenin pathway, associated with human colon tumors [β has been shown as a target for nonsteroidal anti-inflammatory drug- (NSAID-) induced chemopreventive effects in colon cancer [β to bind to its peroxisome proliferator response elements (PPREs) in vitro, while PPARβ overexpression was able to rescue NSAID induced apoptosis in colon tumor cells [β.PPARs play an important role in tumorigenesis and differentiation . PPARα i rodents . PPARβ wn tumors . PPARβ hn cancer , 54. Hign cancer , 149. NSor cells . These fγ has shown promise in therapy promoting terminal differentiation and apoptosis in a variety of malignancies, including liposarcoma, breast cancer, leukemia, gastric cancer, non‐small cell lung cancer, and prostate cancer [γ synthetic ligand troglitazone leads to prostate specific antigen (PSA) stabilization [γ in tumorigenesis. It has been observed that while PPARγ agonists inhibit growth and induce apoptosis in both breast tumor cells and leukemic cells, administration of PPARγ antagonists enhanced this tumor growth inhibitory effect [γ expression [γ agonist ciglitazone induces tumor cell invasion through the generation of ROS and ERK [PPARe cancer –27. In hlization . Althougy effect , 152. Thpression . In fibr and ERK .γ in colorectal tumorigenesis. PPARγ agonists have been shown to promote mouse intestinal tumors, while loss of function mutations of PPARγ has been identified in human colon tumors [γ agonists promote the development of colon tumors in mice with a mutation in the tumor suppressor APC [γ activation by Fas could explain the link between high fat diets and colon cancer. However, one study disputed the PPARγ agonist role in tumor promotion, as it showed that PPARγ agonists were able to induce differentiation and inhibit human tumors from growing in nude mice [γ agonists are able to induce differentiation, cell cycle arrest, and apoptosis in human colon cancer cell lines [γ lead to a reduced number of cancer precursor foci in inflammatory bowel disease [β and PPARγ in colon tumorigenesis may be competition for RXR heterodimerization. The cellular proliferation caused by PPARβ, induced by specific ligands, may lead to overexpression and inhibition of PPARγ heterodimerization, and subsequently contribution to human colon cancer development.This controversy is best exemplified by the role of PPARn tumors , 45, 150ssor APC , 30. Thissor APC . PPARγ aude mice . Furtherll lines . The ant disease . One posγ-associated signaling outcomes. Another example of the differences between species occurs in hepatic cancers. PPARγ agonists are seen as potent carcinogens in rodents, but not seen in humans or primates [γ agonist rosiglitazone seems to enhance carcinogenic effects of the urinary bladder in rodents, while treatment of diabetes with pioglitazone in humans does not seem to increase the incidence of these or any other tumors [These contradictory results might be explained by species specific effects, where a combination of the different factors within each animal leads to different PPARprimates , 158. Fur tumors .γ action are not fully established, it has been shown that PPARγ can inhibit the cell cycle, which is accomplished at least in part through downregulating the protein phosphatase PP2A upon PPARγ activation [γ ligands can also inhibit the G1/S transition by inhibiting Rb phosphorylation [γ upregulates the CDK inhibitors p18 and p21 [γ ligand PGJ2 induces both CDK p21 and the proapoptotic Bax but downregulates the anti-apoptotic Bcl-xL [γ agonist treatment in human pancreatic cancer and bladder cancer cell lines resulted in G1 cell cycle arrest secondary to p21 induction [While the mechanisms underlying PPARtivation . The PPArylation . Further and p21 . PPARγ lc Bcl-xL . Synthetnduction , 165. Fuγ may be explored as a possible intervention in osteosarcoma , which is associated with NSAID-induced apoptosis, has been upregulated by PPARγ in canine OS cell lines [Increasing evidence suggests that activation of PPARosarcoma . PPARγ aeoblasts –10. Therogenesis , 166. A ll lines . The chell lines , 168. KLll lines .γ agonists and 9-cis-retinoic acid have the capability to induce osteoblastic differentiation of OS cells and inhibit OS proliferation [γ to induce apoptosis in chondrosarcoma cells [γ agonist troglitazone promotes the in vitro survival via reduction in apoptosis of the malignant cells [γ and other nuclear receptor hormones. It is plausible that the effects of PPARγ agonists on OS are largely dependent on which step in the differentiation pathway the defect has occurred  and retinoid X receptors (RXRs). RARs are activated by all-trans retinoic acid, a vitamin A metabolite, and heterodimerizes with RXR after ligand binding. RARs have been implicated in early embryonic morphogenesis, including development of the forebrain, hindbrain, and body axis, as well as, early signaling associated with the pancreas, heart, eye, lung, and genitourinary tracts –186. Fur protein , 187–189α induces ligand-independent myogenic differentiation from progenitor cells [RXRs are activated by 9-cis retinoic acid. Beyond the ability of RXR to heterodimerize with many members of the nuclear receptor superfamily, such as PPARs, RXRs can form homodimers. RXRs play important roles in signaling pathways associated with development and carcinogenesis. We have recently demonstrated that exogenous expression of RARor cells . We haveor cells . RXRs aror cells –195. A sor cells –198.γ and retinoids has been carried out in a panel of OS cell lines  has also been shown to play a role in OS cell lines and their responsiveness to therapeutic interventions. VDR is overexpressed on some OS cell lines and administration of 1, 25-dihydroxyvitamin D3 inhibits tumor growth and metastasis, while promoting terminal differentiation , 201. Inγ ligands on different cells in bone and bone marrow microenvironment. Furthermore, these agents can be used as chemopreventive agents for the patients with OS who undergo the resection of the primary bone tumors in order to prevent local recurrence and/or distal pulmonary metastasis. OS is the most frequent primary bone sarcoma, comprising approximately 20% of all bone tumors and about 5% of pediatric tumors overall. OS tumors display a broad range of genetic and molecular alterations, including the gains, losses, or arrangements of chromosomal regions, inactivation of tumor suppressor genes, and the deregulation of major signaling pathways. However, except for p53 and/or RB mutations, most alterations are not constantly detected in the majority of osteosarcoma tumors. Recent studies strongly suggest that OS may be regarded as a differentiation disease that is caused by genetic and epigenetic disruptions of osteoblast terminal differentiation. It has been well established that PPARs and retinoids play an important role in regulating osteogenic differentiation of MSCs. Increasing evidence indicates that PPAR agonists and/or retinoids can inhibit cell proliferation and induce apoptosis in cancer cells, including OS cells. PPAR agonists and/or retinoids may also inhibit OS growth by promoting osteoblastic terminal differentiation. One of the future directions is to develop safe and effective PPAR agonists and/or retinoid derivatives. These agents can be then used as adjuvant therapeutic drugs for OS therapy. Meanwhile, more thorough investigations will be needed to examine the potentially beneficial and/or adverse effects of PPAR


Identifying causal relationships in phylogeographic and landscape genetic investigations is notoriously difficult, but can be facilitated by use of multispecies comparisons.Strix occidentalis caurina; red tree vole: Arborimus longicaudus; southern torrent salamander: Rhyacotriton variegatus; and western white pine: Pinus monticola). Visualizations suggested that, despite occupying the same geographical region and habitats, species responded differently to prevailing historical processes. S. o. caurina and P. monticola demonstrated directional patterns of spatial genetic structure where genetic distances and diversity were greater in southern versus northern locales. A. longicaudus and R. variegatus displayed opposite patterns where genetic distances were greater in northern versus southern regions. Statistical analyses of directional patterns subsequently confirmed observations from visualizations. Based upon regional climatological history, we hypothesized that observed latitudinal patterns may have been produced by range expansions. Subsequent computer simulations confirmed that directional patterns can be produced by expansion events.We used data visualizations to identify common spatial patterns within single lineages of four taxa inhabiting Pacific Northwest forests (northern spotted owl: We discuss phylogeographic hypotheses regarding historical processes that may have produced observed patterns. Inferential methods used here may become increasingly powerful as detailed simulations of organisms and historical scenarios become plausible. We further suggest that inter-specific comparisons of historical patterns take place prior to drawing conclusions regarding effects of current anthropogenic change within landscapes. Landscape genetic and phylogeographic investigations generally rely on a three-step process: data generation, pattern identification, and pattern explanation. Though data generation is generally a straightforward process, the latter two stages can be difficult in many scenarios. Most genetic studies of natural populations produce large, complex data sets . Challenges subsequently ensue when attempting to identify inherent quantitative signals  that provide insights regarding key historical and recent factors that have shaped contemporary genetic structure. Once identified, the set of putative causal mechanisms that may explain said patterns must likewise be discerned. Identification of causal mechanisms becomes notoriously difficult in most cases due to the non-experimental (non-manipulative) nature of investigations and the fact that only a single realization of the evolutionary process can be observed for a given species in a specific geographical region.Despite the challenges associated with identification of causal factors, researchers can nonetheless choose among a wide array of computer programs and analytical procedures that facilitate the inferential process When analyses identify patterns within data sets, causal relationships can nonetheless still be difficult to identify due to the fact that different processes can plausibly produce similar qualitative and quantitative signals In this study, we used a four-stage process to help identify and explain shared patterns of spatial genetic structure in four taxa occupying the same habitat in space and time. First, we used a recently-described visualization technique , a tree vole (Arborimus longicaudus  =  Phenacomys longicaudus), a salamander (Rhyacotriton variegatus), and a tree (Pinus monticola).The goal of this investigation was to explore patterns of spatial genetic structure within single phylogenetic lineages of four different species. Four published data sets were used that met three important criteria: 1) they included reasonable levels of sampling within our region of interest, 2) longitude/latitude or zone 10 UTM coordinates of each sampling location were available, and 3) distinct, single phylogenetic-lineage data sets could be defined for each taxon within the region. Studies meeting these criteria included analyses of a bird . All data for “northern” haplotypes were excluded in our new analyses.Analyses of ng areas . For conp) between individuals for S. o. caurina, A. longicaudus, and R. variegates. For P. monticola, Genetic distances (D) listed in Table 5 of p/(1 – p) or D/(1 – D) and the natural logarithm of geographic distance  in analyses of S. o. caurina, A. longicaudus, and R. variegatus. Nei's genetic distances (D) listed in Table 5 of P. monticola data set. Given that there was variation in the geographical distances between sampling locations (p/(1 – p) or D/(1 – D)] versus the natural logarithm of geographical distances in analyses. Theoretically, this approach accounted for correlations between genetic and geographical distances that may be present and ensured that large interpolation peaks were not resolved solely due to the fact that one or a few sampling areas were geographically isolated Data were subsequently analyzed using the “Genetic Landscape Shape” procedure ocations , we usedOur Genetic Landscape Shape visualizations suggested distinct directional patterns of spatial genetic structure along a North/South axis see . To form−4; δ = 10.0, µ = 10−4; δ = 1.5, µ = 2.5*10−5; δ = 10.0, µ = 2.5*10−5. Note that through manipulation of the above expressions, our dispersal parameters nominally correspond to values of Wright's neighborhood sizes 2) of 18 and 800 for δ  = 1.5 and δ = 10, respectively. Twenty-five simulation replicates of each parameter combination were performed. In the initial phase, simulations were allowed to run for 25,000 generations (µ = 10−4) or 85,000 generations (µ = 2.5*10−5) to permit quasi-equilibrium patterns of spatial genetic structure and diversity to evolve. In the case of a population of 10,000 individuals and µ = 10−4, population genetic theory predicts an expected number of alleles (a) and heterozygosity (H) to be 34.59 and 0.8, respectively, at equilibrium . We followed our normal simulation procedure as described for phase 1, with the exception that empty landscape cells became inhabited during range expansion only when both randomly chosen parental landscape cells were already occupied. For simplicity, we assumed that a newly-occupied cell was always inhabited in all remaining generations. Consequently, over the course of successive simulation generations, the remainder of the landscape was filled at a rate associated with values of δ specified during simulation runs. At two time points during the range expansion event, data sets comprised of a single haplotype from 200 randomly selected individuals were exported for analysis. Ultimately, we examined time points where 1) the lower two-thirds of the landscape (a 100×200 region) was inhabited, and 2) the landscape was completely inhabited. On average, simulations using δ  = 1.5 required 424.21 generations (S.D. = 6.81) and 824.53 generations (S.D. = 12.53) to reach the two-thirds full and completely inhabited states, respectively. Likewise, simulations using δ  = 10 required 62.63 generations (S.D. = 1.07) and 110.78 generations (S.D. = 1.44) to reach the two-thirds full and completely inhabited states, respectively.Once data were generated, AIS was used to analyze simulated data sets using the Genetic Landscape Shape procedure. Resulting surface plots revealed directional patterns of spatial genetic structure where inter-individual genetic distances were smaller in the expansion zone relative to the southern refugial region see . ConsequA. longicaudus and R. variegatus . Likewise, spatial autocorrelations illustrated that pairwise genetic distances were significantly smaller than average over shorter distances and were significantly larger than random expectations as geographic distances increased. In contrast, both analyses suggested that no  spatial genetic structure was present for S. o. caurina and P. monticola  were present in the south relative to the north. In contrast, A. longicaudus and R. variegatus indicated opposite trends. Our post-hoc analyses illustrated significant associations between UTM northing and genetic distances in all four species . In contrast, correlations for A. longicaudus and R. variegatus were positive .Genetic Landscape Shape visualizations suggested two primary patterns of directional spatial genetic structure and diversity . In S. o species . Consist−4) relative to low (µ = 2.5*10−5). Finally, in three of the four parameter combinations, more significant directional patterns were observed after the landscape became completely inhabited relative to the two-thirds inhabited case.Simulated range expansions revealed directional patterns whereby inter-individual genetic distances were reduced in the direction of the expansion event , Table 1Most population genetic and phylogeographic investigations produce large, complex data sets. For example, the (n*(n-1))/2 genetic distances that can be calculated from all pairwise combinations of n samples or sampling locations may generate extremely unwieldy matrices that are difficult to interpret, especially when there is an interest in identifying patterns based on joint analyses of spatial and genetic information. Historically, such data sets have been analyzed using Mantel tests A. longicaudus and R. variegatus showed strong spatial genetic structure, whereas P. monticola and S. o. caurina revealed no (or little) evidence of such patterns. When visualizations were implemented, an alternate view of spatial genetic structure within these species emerged. Interestingly, the overt patterns observed were of a type not generally considered in most investigations: shared directional patterns of genetic structure existed along a South to North transect . In a regional context, this pattern reflected greater genetic diversity in southern relative to northern locales. In contrast, genetic distances were larger in the North relative to the South for A. longicaudus and R. variegatus. Although visualizations are qualitative descriptions of spatial genetic structure patterns, additional post-hoc statistical analyses confirmed the graphical depictions generated  likely avoided cold temperature regimes during glacial periods by actively moving to more suitable southern refugia. As suggested for other avian species S. o. caurina likely expanded its range northward following glacial retreat once environmental conditions became favorable. Similarly, we suggest that P. monticola, while clearly not capable of avoiding unfavorable climatic conditions, likely was instead extirpated from northern regions of its range during glacial periods. Again, following glacial retreat, the species may have been reestablished in northern areas through wind and animal-mediated seed dispersal. P. monticola seeds generally disperse <120 m from parental trees, but have been reported to travel upwards of 800 m in some cases Pinus species, mammal and bird species actively feed upon, cache, and disperse P. monticola seeds S. o. caurina and P. monticola, we suggest that this process may have disrupted a previously-existing isolation-by-distance process and lead to the inability of these two procedures to detect underlying patterns.Though our empirical data reflected the same general geographical area, our analyses indicated that different species responded differently to the same historical events and processes that occurred.  results , as bothR. variegatus and A. longicaudus displayed patterns opposite to those identified in S. o. caurina, P. monticola, and the strict northward range expansion scenario encapsulated in simulations  is protected under the U.S. Endangered Species Act as a threatened subspecies A. longicaudus is currently the only mammal protected by the “Survey and Manage” portion of the Northwest Forest Plan due to its importance as a food source for S. o. occidentalisR. variegatus was previously petitioned, but deemed not warranted, for listing under the U.S. Endangered Species Act P. monticola and A. longicaudus, respectively (estimated from r2 values obtained from squaring correlation coefficients; Three of the four species investigated are of conservation importance in the Pacific Northwest United States. The northern spotted owl (Debates over the most appropriate tools for genetic studies of natural populations are not likely to end in the near future, particularly when undertaking the challenge of comparing multiple species. We therefore agree with points raised by others regarding use of different integrative analytical tools in a heuristic manner when making inferences


Feedback postural control depends upon information from somatosensation, vision, and the vestibular system that are weighted depending on their relative importance within the central nervous system. Following loss of any sensory component, the weighting changes, e.g., when suffering a vestibular loss, the most common notion is that patients become more dependent on visual cues for maintaining postural control. Dizziness and disequilibrium are common after surgery in schwannoma patients, which could be due to interpretation of the remaining sensory systems involved in feedback-dependent postural control and spatial orientation.  To compare visual dependency in spatial orientation and postural control in patients suffering from unilateral vestibular loss within different time frames. n = 27) with no vestibular function prior to surgery (lost through years), group 2 (n = 12) with remaining vestibular function at the time of surgery (fast deafferentation), and group 3 (n = 18) with remaining function that was lost through gentamicin installations in the middle ear (slow deafferentation). All patients performed vibratory posturography and rod and frame investigation before surgery and 6 months after surgery.  Patients scheduled for schwannoma surgery: group 1 (p < 0.001). Patients that suffered fast loss of remaining vestibular function (group 2) became less visual field dependent after surgery (p ≤ 0.035) and were less able to maintain stability compared with group 1 (p = 0.010) and group 3 (p = 0.010).  Postural control improved after surgery in patients that suffered a slow deafferentation (groups 1 and 3) ( The nature and time course of vestibular deafferentation influence the weighting of remaining sensory systems in order to maintain postural control and spatial orientation. Postural control is maintained by both feedback and feed-forward mechanisms . Feed-foAt our hospital, we do a thorough vestibular work-up prior to surgery with the aim to determine what function remains. If significant vestibular function remains, the patients are offered our pretreatment program (“PREHAB”) . PREHAB Postural control is most often evaluated by posturography, and it has been shown that the control system needs to be sufficiently challenged during any evaluation of sensory preference, since the involved sensory systems overlap each other in terms of motion detection and spatial orientation . As for The aim of the present study was to compare visual dependence both with regard to postural control evaluation and spatial orientation in patients suffering unilateral vestibular loss with different time frames.Between the years 2002 and 2011, 136 patients were subjected to vestibular schwannoma surgery at Lund University Hospital. Most patients underwent an initial assessment, in which the vestibular function was investigated with video-recorded head impulse test of all 3 canals of each ear, bithermal calorics, vestibular-evoked myogenic potential measured on the sternocleidomastoid muscle (cVEMP), subjective horizontal and vertical, rod and frame tests, posturography, and eye movement analyses. Most patients also came to a follow-up after 6 months, in which posturography, video-recorded head impulse test, eye movement analyses, and rod and frame test were performed. The criteria for the present study were performed posturography and the rod and frame test, both at the initial assessment and the postsurgical follow-up. There were no signs of central dysfunction  either before or after surgery, since central nervous dysfunction could confound the results. Full records could be retrieved in 57 patients. The reason for incomplete data were CNS affection in 35 patients, 14 patients did not perform rod and frame prior to surgery as it was not a set routine, 13 declined follow-up due to them living far away, 12 patients had incomplete data due to computer failure or move of laboratory, 2 were diagnosed with malignancies and declined follow-up, 1 patient was not able to perform posturography after surgery with eyes closed (group 2 below), 1 patient did not tolerate vibration to the legs before surgery, and 1 did not perform the correct posturography test.The final 57 patients were divided into 3 groups according to vestibular function before surgery and if they performed the PREHAB program.no vestibular function” consisted of patients with no detectable vestibular function prior to surgery, i.e., best caloric response <7.5°/s, pathological head impulse tests of the affected ear, and no cVEMP responses. 17 were females and 10 were males, age 58.2 ± 9.6 years. 19 were subjected to translabyrinthine extirpation and 8 to retrosigmoidal approach. Individual vHIT performance was assessed by an experienced neurootologist, and a pathological impulse was defined as a gain <0.6 and presence of catch-up eye saccades (either overt or covert). Tumor extrameatal size was 20.7 ± 8.4 mm.Group 1  “vestibular function” consisted of patients with remaining vestibular function prior to surgery. These patients were not treated with gentamicin either because they were diagnosed before 2004  or because their hearing was completely or near to normal and hence candidates for hearing preserving surgery (n = 7). 3 were females and 9 were males, age 47.8 ± 10.6 years. 4 were subjected to translabyrinthine extirpation and 8 to retrosigmoidal approach. Tumor extrameatal size was 17.3 ± 8.1 mm.Group 2 (12 patients) “PREHAB” consisted of patients with remaining vestibular function prior to surgery that were treated with intratympanic gentamicin installations [Group 3 (18 patients) “llations . The genThe decision of surgical approach was based upon location and size of the tumor and whether hearing preservation was attempted. Hearing preservation also excluded patients from gentamicin treatment, i.e., hearing mean threshold <30 dB nHL  and speech discrimination >70%.The study (2014/171) was approved by the local ethical board (EPN) at Lund University, Sweden, and the study was performed in accordance with the declaration of Helsinki, with patients giving their informed consent.Postural control was evaluated by perturbing stance while standing on a force platform (400 × 400 × 75 mm) equipped with six strain-gauge sensors. Forces and torques actuated by the feet were recorded with six degrees of freedom by a force platform. Data were sampled at 50 Hz by a computer equipped with a 16-bit AD converter. Postural perturbations were induced by simultaneous vibratory stimulation to the belly of the gastrocnemius muscles of both legs  VibratioThe vibration amplitude was 1.0 mm amplitude at a constant frequency of 85 Hz. The vibratory stimulation was executed according to a computer-controlled pseudorandom binary sequence (PRBS) schedule  for 205 After informing about the test procedure, the subjects were instructed to stand erect but not at attention, with arms crossed over the chest and feet at an angle of about 30 degrees open to the front and the heals approximately 3 cm apart. Two tests were conducted at each trial occasion, eyes open, fixating on mark on the wall at a distance of 1.5 m, and eyes closed. The test order followed our set clinical procedure, always starting with eyes open followed by eyes closed. In order to minimize external disturbances, the test subjects listened to classical music relayed through headphones .The rod and frame test was performed in a dark room, with the subjects seated with their heads immobilized with straps against a neck rest. On a wall 1.5 m in front of them, a 15 cm long and 2 mm wide dimly lit light bar (rod) was projected, which remotely by the patients could be rotated in the frontal plane. The instructions were to align the rod horizontally, either with no other visual influences or with the rod surrounded with a tilted frame . The test procedure was set, starting with no frame, followed by frame tilt. Four measurements each were made of the subjective horizontal and in the frame, and the mean was calculated. The test had no time limit.The variance in recorded torque was calculated for quiet stance (0–30 seconds) and for the stimulation period 31-230 seconds). The data were normalized by squared mass and squared height since regression analysis showed dependence on those factors , 23. Vib0 secondsPostural stability while standing is commonly analyzed using force platforms and the movements of the center of pressure (CoP), i.e., the point of application of the ground reaction force. We present torque variance values from the force platform recordings because these values correspond directly with the energy used towards the support surface, and changes in recorded torque from the force platform correspond well to the actual body movements and posture changes induced by vibratory stimulus . HoweverA ratio close to zero or negative indicates that the stability was similar or poorer with EO than with EC, i.e., visual information was less important for postural control. This formula considers the total amount of body sway during both visual conditions (EO and EC).The rod and frame test values were normalized to the side of the tumor  in test with no frame, and in the Frame test to which side the frame was tilted, i.e., if the frame tilt was tilted to the side of the tumor  or to the opposite side  .The anteroposterior torque variance values during quiet (unperturbed) stance and during balance perturbations were analyzed using repeated measures GLM ANOVA  on log-transformed values . The maiU (exact sig. 2-tailed) test was used for between-groups post hoc comparisons. The Wilcoxon matched-pairs signed-rank test (exact sig. 2-tailed) was used for within-subjects post hoc comparisons, i.e., analyzing the changes between before and after treatments.The Mann-Whitney p values < 0.05 were considered statistically significant. For the Mann-Whitney comparisons, the significant Bonferroni corrected level was set to p < 0.025. For the Wilcoxon post hoc comparisons, the significant Bonferroni corrected level was set to p < 0.05. Nonparametric statistical tests were used in all statistical evaluations since the Shapiro-Wilk test revealed that some of the obtained datasets were not normally distributed and normal distribution could not be obtained after log transformation.In all GLM ANOVA analyses, p < 0.001) (p = 0.017).During quiet stance, visual cues improved the stability in all 3 groups (< 0.001) . Howeverp < 0.001). Postural control improved in the “PREHAB” and the “no vestibular function” groups after surgery (p < 0.001). Moreover, the “PREHAB” group had generally better stability before and after surgery than the “no vestibular function” group (p = 0.002). Furthermore, both the “PREHAB” (p = 0.010) and “no vestibular function” (p = 0.010) groups had significantly better stability postsurgery than the “vestibular function” group.During vibratory perturbation, visual cues improved the stability in all patient groups (p = 0.043).None of the groups significantly changed their quiet stance stability between before and after surgery with eyes closed or eyes open . Howeverp ≤ 0.018) and “PREHAB” (p ≤ 0.016) groups significantly increased performance both with eyes closed and eyes open between before and after surgery. With eyes closed, the “PREHAB” group presented trends of better stability than the “no vestibular function” group (p = 0.032) before surgery. After surgery, the “PREHAB” group had significantly better stability than the “no vestibular function” group (p = 0.024) and the “vestibular function” group (p = 0.017). With eyes open, trends suggest poorer stability in the “no vestibular function” group compared to the “vestibular function” (p = 0.031) group and the “PREHAB” (p = 0.031) group. After surgery, the “PREHAB” group had significantly better stability than the “no vestibular function” group (p = 0.022) and a trend suggests better stability than the “vestibular function” group (p = 0.035).During vibratory perturbation, the patients with remaining vestibular function before surgery did not improve postural performance between before and after surgery with eyes closed and eyes open . Howeverp = 0.018) (The Romberg ratio during quiet stance was significantly higher in the “vestibular function” group compared with “no vestibular function” group (= 0.018) . During p = 0.042) (The Romberg ratio in quiet stance showed a trend to be higher in the “vestibular function” group compared with the “no vestibular function” group postsurgery (= 0.042) .During vibratory stimulation, there were no significant differences between the groups or before and after surgery . There sp ≤ 0.049) (p = 0.004) and significantly larger visuospatial errors postsurgery in the “PREHAB” group than in the “no vestibular function” group (p = 0.027). Ipsilesional frame effect was significantly lower in the “vestibular function” group compared with the “no vestibular function” group (p = 0.027) and the “PREHAB” group (p = 0.042). Contralesional frame effect was significantly lower in the “vestibular function” group compared with the “no vestibular function” group (p = 0.006) and the “PREHAB” group (p = 0.014).The error in perceived earth horizontal increased after surgery in all GLM ANOVA group constellations (≤ 0.049) . Howeverp = 0.010) and “PREHAB” (p = 0.013) groups than in the “no vestibular function” group (p = 0.026) and the “PREHAB” group (p = 0.035). Finally, the contralesional frame effect was significantly smaller in magnitude after surgery in the “vestibular function” group compared with the “no vestibular function” group (p = 0.002) and the “PREHAB” group (p = 0.013). Furthermore, a trend suggests a smaller contralesional frame effect before surgery in the “vestibular function” group compared with the “PREHAB” group (p = 0.048).The error in perceived earth horizontal was smaller before surgery in the “vestibular function” (n” group . Moreovep = 0.012) and “PREHAB” (p = 0.003) groups than after surgery.The error in perceived earth horizontal was significantly smaller before surgery in the “vestibular function” . Those with “no vestibular function” lost the function probably over years due to the slow tumor growth, and the “PREHAB” group over weeks, and those with remaining vestibular function at the time of the surgery as the nerve was cut. The differences between the groups could be summarized as better postural performance with slow deafferentation and less visual field dependency with fast deafferentation. There also seemed to be a tendency with fast deafferentation for less use of visual cues to stabilize posture after surgery, which has been demonstrated earlier . These fp = 0.043 vs PREHAB) and swayed more with both eyes closed and eyes open during challenging stability perturbations after surgery  aim at inducing motor learning at a cellular level before the vestibular loss, in line with multiple plasticity mechanisms active in cerebellar and hippocampal adaptation –38, as wSeparating the sensory trauma (uVD) from the surgical trauma in the PREHAB program might be beneficial for compensation processes in general. It is well known that surgery interferes with central structures such as hippocampal function , 45 and It seems that the different time courses of unilateral vestibular deafferentation affect postural competence, and these performance differences could be the consequence of different sensory weighting differences. This should be considered when designing rehabilitation programs for patients suffering from uVD. Visual dependency or not does not necessarily dictate postural performance nor dizziness, but in order to help each patient in their rehabilitation, it may be of importance to assess the individual sensory components of the postural control system and spatial orientation.


There is an urgent need for novel, noninvasive biomarkers to diagnose Alzheimer’s disease (AD) in the predementia stages and to predict the rate of decline. Therefore, we set up the European Medical Information Framework for Alzheimer’s Disease Multimodal Biomarker Discovery (EMIF-AD MBD) study. In this report we describe the design of the study, the methods used and the characteristics of the participants.Participants were selected from existing prospective multicenter and single-center European studies. Inclusion criteria were having normal cognition (NC) or a diagnosis of mild cognitive impairment (MCI) or AD-type dementia at baseline, age above 50 years, known amyloid-beta (Aβ) status, availability of cognitive test results and at least two of the following materials: plasma, DNA, magnetic resonance imaging (MRI) or cerebrospinal fluid (CSF). Targeted and untargeted metabolomic and proteomic analyses were performed in plasma, and targeted and untargeted proteomics were performed in CSF. Genome-wide SNP genotyping, next-generation sequencing and methylation profiling were conducted in DNA. Visual rating and volumetric measures were assessed on MRI. Baseline characteristics were analyzed using ANOVA or chi-square, rate of decline analyzed by linear mixed modeling.n = 492, MCI n = 527, AD-type dementia n = 202) with a mean age of 67.9 (SD 8.3) years. The percentage Aβ+ was 26% in the NC, 58% in the MCI, and 87% in the AD-type dementia groups. Plasma samples were available for 1189 (97%) subjects, DNA samples for 929 (76%) subjects, MRI scans for 862 (71%) subjects and CSF samples for 767 (63%) subjects. For 759 (62%) individuals, clinical follow-up data were available. In each diagnostic group, the APOE ε4 allele was more frequent amongst Aβ+ individuals (p < 0.001). Only in MCI was there a difference in baseline Mini Mental State Examination (MMSE) score between the A groups (p < 0.001). Aβ+ had a faster rate of decline on the MMSE during follow-up in the NC (p < 0.001) and MCI (p < 0.001) groups.We included 1221 individuals  contains supplementary material, which is available to authorized users. Over the last decade great progress has been made in diagnosing Alzheimer’s disease (AD) at an early disease stage, including before the onset of dementia , 2. The Currently, CSF and PET biomarkers are commonly used for the early diagnosis and prognosis of AD –9. Howevhttp://www.emif.eu). Funded through the Innovative Medicines Initiative (IMI), the EMIF project was established to facilitate the process of reusing and combining existing healthcare data with a focus on two therapeutic areas in the first instance: metabolic diseases and AD. One of the main aims of the EMIF-AD project is to accelerate the discovery of novel diagnostic and prognostic biomarkers for AD and to unravel the underlying pathophysiological mechanisms, using existing data and existing samples, that would otherwise be inaccessible to research beyond the project teams responsible for the collection. In this report, we will describe the set-up of the EMIF-AD Multimodal Biomarker Discovery (EMIF-AD MBD) study, the methods as well as the characteristics of the included subjects. The results of the single and multimodal analyses will be described in future publications.This study was performed as a part of the European Medical Information Framework for Alzheimer’s disease , 400 subjects with mild cognitive impairment (MCI) and 200 subjects with mild AD-type dementia. To create a balanced design in terms of progression and to enable endophenotype designed biomarker studies, we intended to include 50% Aβ-positive (Aβ+) individuals and 50% Aβ-negative (Aβ–) individuals in the groups with NC and MCI. To conduct multimodal analyses, we initially aimed to include subjects who had material from MRI, plasma, DNA and CSF. Later, we adjusted this to subjects with material available in at least two of the modalities listed.https://emif-catalogue.eu), established as part of the objective of the EMIF which seeks to enable the finding, assessment and reutilization of preexisting data. The EMIF-AD Catalog contains metadata about European AD cohorts, enabling the selection of studies that included subjects who, in this instance, met the following inclusion criteria: data on Aβ status, measured in CSF or by amyloid positron emission tomography (PET); age above 50 years at baseline; and availability of MRI scans, plasma and DNA samples. We identified 16 suitable cohorts. Two cohorts declined due to other research interests. Three cohorts were interested to collaborate, but unable because of legal and/or ethical restrictions, or unavailability of sufficient sample volumes. The 11 selected cohorts included three multicenter studies—EDAR (n = 204) [n = 147) [n = 29) [n = 149) [n = 172) [n = 45) [n = 120) [n = 180) [n = 40) [n = 95) [n = 40) [We used the EMIF-AD Catalog n = 204) , PharmaCn = 147)  and DESCn = 149) , Amsterdn = 172) , Barcelo[n = 45) , Barcelo4 , Pharmn = 180) , San Seb[n = 40) , Gothenb[n = 95)  and Laus[n = 40) . Of thesIn all cohorts, the definition for NC was a normal performance on neuropsychological assessment . Five cohorts also used a score of 0 on the Clinical Dementia Rating (CDR)  and a siAll cohorts were asked to contribute available data on demographics, clinical information, neuropsychological testing and Aβ status, as presented in Table n = 1216). We requested at least one test from the following cognitive domains: memory, language, attention, executive functioning and visuoconstruction [z-scores calculated based on local normative data. If local normative data were unavailable, we calculated z-scores based on published normative data from healthy controls for that test. Per cognitive domain, we combined z-scores which we used as a continuous variable, and we used a cutoff value of z-score < − 1.5 to define abnormality.Cognitive data were collected in all cohorts. The cognitive tests used varied across centers. Only the Mini Mental State Examination (MMSE) was administered in all centers and was available for nearly all subjects n = 12. We requClinical data were harmonized, pooled and stored on an online data platform using tranSMART , now enrInitially, the minimum requested amount of plasma was 0.7 ml. If available, another 0.7 ml was requested to conduct additional analyses in a subgroup of subjects. In some cases, only 0.5 ml was available. Prior to the analyses, samples were checked visually for consistency and volume. Results of analyses were also quality checked by inspecting patterns of outliers, and excluding consistent outliers across analyses. Analyses conducted on these samples include: targeted analyses of plasma proteins identified previously  and confA total amount of 2.6 μg DNA or 1 ml whole blood, from which DNA was to be extracted, was requested for the genetic analyses. After performing routine quality checks on extracted DNA , we performed three types of assessments on each sample passing quality control: genome-wide SNP genotyping , genome-wide DNA methylation profiling  and whole exome sequencing.38, Aβ40, Aβ42, Aβ42/40, YKL-40, NFL and neurogranin (Ng). Prior to the analyses, CSF samples were checked visually for volume and absence of blood contamination.The requested amount of CSF to conduct all planned analyses was 0.4 ml, which was used for untargeted proteomic and peptidomic analyses, and a number of targeted analyses measuring concentrations of Aβhttps://neugrid4you.eu) [MRI scans were assembled centrally, quality checked and assessed visually by a single rater. T1-weighted and, when available, FLAIR and/or T2*/SWI images were used for qualitative visual rating, including medial temporal lobe atrophy , global 4you.eu)  for stor42/40 of the central analyses, using a cutoff value of < 0.061 to determine abnormality (n = 770). The cutoff value for the Aβ42/40 ratio was determined based on mixture model analyses comparing the NC and AD groups in this dataset. When no CSF was contributed for central analyses, the local CSF Aβ42 value (n = 271) or the standardized uptake value ratio (SUVR) on an amyloid PET scan (n = 180) with local cutoff values to determine abnormality were used  and years of education (n = 105) were imputed using regression within study with at least two significant predictors . Statistical analyses were performed using R Statistical Software (version 3.3.3) and SPSS (version 24), with significance defined as p < 0.05.Baseline characteristics were compared between groups using ANOVA for continuous variables and chi-square for categorical variables. General linear mixed models with random intercepts and slopes by study were used to examine the influence of Aβ status on MMSE performance and decline over time, adjusted for age, gender and years of education. Missing values for APOE genotype  years. Six hundred and sixty-seven (54%) were female and the average education level was 11.7 (SD 4.1) years. At baseline, 492 (40%) subjects had NC, 527 (43%) subjects had a diagnosis of MCI and 202 (17%) subjects had a clinical diagnosis of AD-type dementia. For 758 (62%) individuals there were follow-up data available : 217 (44%) NC subjects, 398 (76%) MCI subjects and 143 (71%) demented subjects. The average follow-up time for all 758 individuals was 2.3 (SD 1.2) years. Per diagnostic groups, the average clinical follow-up time was: NC 2.4 (SD 0.9) years, MCI 2.2 (SD 1.3) years and AD 2.2 (SD 1.4) years.p = 0.002; MCI, p < 0.001). In all diagnostic groups, Aβ+ subjects were more likely to be an APOE ε4 carrier . In the MCI subjects only, there was a difference in baseline MMSE score between the Aβ groups (p = 0.001). Regarding cognitive domains, we found differences in memory (p < 0.001) and executive functioning (p = 0.042) z-scores in individuals with MCI. In individuals with AD-type dementia we found that Aβ+ individuals performed worse on an executive functioning task (p = 0.013).Table Table p < 0.001). In the NC and MCI groups, the Aβ+ individuals in the NC and MCI groups decline at a faster rate than the Aβ– individuals . For the demented subjects, Aβ did not influence the rate of decline  to work with these data, which require approval from the EMIF-AD team and the data-owners.AD is a complex and multifactorial disorder, which underscores the need for multimodal studies with sufficient statistical power . To dateBesides the major advantages, this study also has some limitations. Currently, we do not have clinical follow-up data for all subjects, as some centers are still in the process of collecting these. However, these data may be added to the database in the future. Also, the data, samples and scans contributed to this study were collected at different centers and were not collected using the same protocol, which will lead to preanalytical variability. To limit this variability, the samples were analyzed centrally and the clinical data were harmonized using standardized values and dichotomous variables.The various complementary analyses conducted in plasma, DNA and CSF and on MRI scans in a large-sized cohort of individuals across the clinical AD spectrum provide a unique opportunity to discover novel diagnostic and prognostic markers, and will also increase knowledge into the AD pathophysiology, which is required for the development of novel therapeutic interventions.Additional file 1:Table S1. Diagnostic criteria per cohort. Cohorts, countries, number of subjects and diagnostic criteria used for NC, MCI and AD dementia (DOCX 90 kb)Additional file 2:Table S2. Number of subjects per test by cognitive domain at baseline. Number of subjects per test and norms used in domains of global cognition, memory, language, attention, executive functioning and visuoconstruction (DOCX 90 kb)Additional file 3:Table S3. Biomarker protocol information for each cohort. Protocol information for PET, CSF and plasma biomarker collection (DOCX 100 kb)Additional file 4:Table S4. Ethical approval committee of each center. Ethical approval committees in each of the participating centers (DOCX 109 kb)


We present the case of a 68-year-old male, who underwent open abdominal aortic graft in August 2016 owing to a ruptured large infrarenal abdominal aneurysm. He subsequently presented 6 months later with back pain, general weakness, reduced mobility and cachexia. He underwent CT, MRI and fluorodeoxyglucose (PDG)-PET spinal imaging, all modalities showing signs of aortic graft infection complicated by L4/5 discitis. The patient was treated conservatively with intravenous antibiotics and spinal brace support, as his general condition did not allow for surgery. Although he showed initial clinical improvement allowing plans for supported discharge, his improvement was not sustained and he died 4 months after admission. A 68-year-old male presented as an emergency in August 2016 with a diagnosis of a ruptured infrarenal abdominal aortic aneurysm. He underwent emergency open vascular surgery with the placement of an aortic bifurcated graft with intraoperative right femoral embolectomy. One day post-operatively, he required a left femoral embolectomy. In February 2017—6 months after his surgery—he presented through the emergency department with worsening low back pain, progressive fatigue, decreased appetite and oral intake. He was severely cachexic. His notable comorbidities included chronic renal failure and had a past history of excess alcohol intake. On admission, he was apyrexial (36.7 C), normotensive (BP 110/70 mmHg) with a normal pulse rate (68 per min). He was severely cachectic weighing 37 kg with a body mass index of 12.4. On palpation of the lumbar spine, there was tenderness without radiation and there were no focal neurological signs in the lower limbs. His breathing sounds were clear and his abdomen was soft with a healthy midline scar of his previous surgery.The blood results were as follows:−1, (130–170 g l−1) white cell count 10.2 10*9 l−1 (4.0–11.0 × 10*9 l−1) , platelets 462 10*9 l−1 (150–400 10*9 l−1). The C-reactive protein (CRP) was elevated at 64 mg l−1 (0–9 mg l−1). His liver profile showed an elevated alanine transaminase level of 554 IU l−1 (0–35 IU l−1) and alkaline phosphatase 1446 IU l−1 (30–150 IU l−1), gamma-glutamyl transferase 670 IU l−1 (9–44 IU l−1), serum total bilirubin 9 umol l−1 (0–17 umol l−1), prothrombin time 11.1 s. (9.9–12.8 s), fibrinogen 8.10 g l−1 (2.00–5.10 g l−1), international normalized ratio 1.0 (0.9–1.2) and albumin 39 g l−1 (35–48 g l−1), serum amylase 70 IU l−1 (0–100 IU l−1), plasma glucose 7.1 mmol l−1 (4.0–7.7 mmol l−1). His corrected calcium was elevated at 2.76 mmol l−1 (2.15–2.60 mmol l−1) with a normal parathyroid hormone level of 3.6 pmol l−1 (1.9–6.4 pmol l−1). Electrolytes and kidney function test showed hyperkalaemia 6.7 mmol l−1 (3.5–5.0 mmol l−1) and mild hyponatraemia with a sodium level of 131 mmol l−1 (132–146 mmol l−1) with a kidney injury; creatinine 267 umol l−1 (59–104 umol l−1), urea 37.3 mmol l−1 (2.5–6.7 mmol l−1) and estimated glomerular filtration rate 21 ml min−1/1.73 m−2.Haemoglobin 120 g l Patient was initially managed with intravenous fluids along with optimization of his nutrition including nasogastric feeding with regular follow up by dieticians. A treatment with intravenous pamidronate was initiated to correct hypercalcaemia. A liver screen including a viral hepatitis screen was negative and a magnetic resonance cholangiopancreatography showed a dilated common bile duct and intrahepatic ducts. A prostate specific antigen was done to rule out possible prostate malignancy and bone metastasis. Myeloma screen was negative. Flexible sigmoidoscopy showed a benign looking polyp with no ulcers or any evidence of malignancy. An abdominal ultrasound showed normal kidneys and spleen structures and a thrombus in the aortic aneurysm. Later, CT chest, abdomen and pelvis showed soft tissue posterior to the aneurysm as well as periaortic inflammatory process, no malignancy was found . A diagn−1, CRP of 199 mg l−1. Although his CRP was initially falling down in the first few days, patient’s back pain was progressive and his WBC and CRP climbed up markedly . Of note, patient did not develop high temperature—not once—during his hospital stay. A blood culture was done, one bottle showed positive growth of coagulase negative Staphylococcus. However, repeated blood culture after 48 h was negative. After 6 weeks of admission, patient developed hospital-acquired pneumonia and was started on IV tazocin. His bloods showed elevated white blood cells (WBC) of 14.00 10*9 l An MRI (lumbar spine) showed a small abscess collection over the right paravertebral region . Radiolo However, follow-up CT (abdomen/pelvis) was done after 10 weeks of antibiotic treatment and showed no improvement of previous findings. Moreover, an FDG-PET scan of lumbar spine showed an increased uptake in L4/5 disc with continuity to the right psoas muscle .−1, 73 mg l−1 respectively). A repeated blood culture was negative. Owing to the progressive decline in the patient’s general status, despite a long antibiotic course of 15 weeks, he was commenced on an end of life care and died in early July 2017. Soon patient’s condition worsened with a progressive back pain rendering him bedridden. He developed low body temperature of 34.8°C and his WBC and CRP reached 4 is caused by low-virulent bacteria and occurs generally late after the original graft implantation. Usually, these patients suffer from non-specific symptoms such as weakness, weight loss, and malaise; potentially leading to delayed diagnosis. Our patient had non-specific symptoms related to ongoing low-grade infection. Owing to the fact that his blood cultures were negative on two separate occasions and patient refused sampling from the infection field, it was mandatory to ascertain the diagnosis using different radiological investigations. For a proper diagnosis, it is mandatory to do clinical examination, blood tests  and radiological investigations. As cultures of blood or even samples collected from the infected field are sometimes negative (up to 33% of cases), CT scan is the most commonly used modality for the diagnosis of aortic graft/aneurysmal wall infections. Potential signs of graft infection on CT scan are perigraft fluid, perigraft soft tissue attenuation, ectopic gas and pseudoaneurysm. Yet, perigraft fluid and soft tissue attenuation are not specific for graft infection, as they might arise from chronic postoperative inflammatory process. Therefore, MRI scan has an additional value of a more accurate evaluation of soft tissue changes.7 A grading system was adopted for PDG-uptake, the intensity of which was graded on a 5-point scale, as follows: Grade 0, FDG uptake similar to that in the background; Grade 1, low FDG uptake, comparable to that by inactive muscles and fat; Grade 2, moderate FDG uptake, clearly visible and distinctly higher than the uptake by inactive muscles and fat; Grade 3, strong FDG uptake, but distinctly less than the physiological uptake by the bladder; and Grade 4, very strong FDG uptake, comparable to the physiological urinary uptake by the bladder.7 In our case, patient showed a high density uptake in L4\5 disc, focal uptake in the anterior part of the aneurysm  and the paraspinal collection that would match Grade 4–5 in this system. The direct extension of the aneurysmal wall into the disc with the presence of the paraspinal collection seen in the MRI scan favours the diagnosis of infection rather than the mere presence of the postoperative changes in the area of interest. FDG-PET scan is nowadays commonly used to diagnose various infections. Fukuchi et al described a high specificity of FDG-PET in the diagnosis of aortic graft infections when certain criteria of FDG-uptake pattern is met.staphylococcusaureus was found to be a common cause of infection; in 54.5% of the cases as reported by Ducasse et al,8 patient was treated with intravenous teicoplanin and rifampicin. The source of infection was unknown in this case although the intra- and post-operative embolectomy interventions are thought to have probably introduced graft infections. As 10 Sharif et al described a 100% mortality rate in conservatively managed patients.9 Heyer et al reported 75%,10 Ducasse et al reported 36.4%.8 Our patient had acute on top of chronic renal failure, which worsened the prognosis. Unfortunately, he died after conservative treatment failed to control his infection. According to multiple studies, outcome markedly worsens when conservative management is used.Aortic aneurysm graft infections are rare and could be misdiagnosed, unless there is an early index of suspicion.Presentation of infected aortic grafts varies and could be subtle with general non-specific symptoms which may delay initiation of proper treatment.CT, MRI and PET-CT scan has an increasing role in the diagnosis of graft infections and should be considered early in the course as laboratory evidence of infection might not be clear enough to raise the alarm for a prompt antibiotic therapy. Therefore, it is mandatory to carefully check for any signs of discitis in the CT scan and if there is a suspicion, a spine MRI should be immediately recommended.


Class 2 patients  had profiles in between the other two classes. Additionally, Class 1 and 2 patients had better quality of life than Class 3 patients. Our results suggest that promoting light PA and altering perceived body shape through counselling may improve quality of life in patients with dyslipidemia.Despite the increasing prevalence and economic burden of dyslipidemia in South Korea, we have little data on the physical activity of patients. Thus, we aimed to investigate how quality of life among patients with dyslipidemia is influenced by a combination of the following variables: light physical activity (PA), sedentary behavior (SB), perceived body shape, and body mass index (BMI). We examined data from the Sixth Korean National Health and Nutrition Examination Survey (KNHANES VI 2015), collected in 2015 by the Korean Centers for Disease Control and Prevention. The analysis included 534 individuals with dyslipidemia out of 7380 survey participants. Latent profile analysis identified three latent classes of individuals based on their physical profiles. Class 1 patients (active;  It is a007,318) . Medical007,318) . BecauseA few encouraging signs for this association include the finding that light PA  decreases dyslipidemia indicators such as TC and high-density lipoprotein (HDL) cholesterol ,10. In cQuality of life has psychological, physical, and social dimensions . The conLatent profile analysis (LPA) is a person-centered approach that should be effective for exploring subtypes of patients with dyslipidemia. The method assumes that patients can be classified into latent groups based on a set of characteristics (their profiles) . This peIn this study, we aimed to first investigate whether South Korean patients with dyslipidemia can be classified into latent classes based on light PA, SB, BMI, and perceived body shape. Next, we aimed to examine how these latent classes influenced quality of life. Our results should provide insight on key correlates of quality of life among patients with dyslipidemia, thus helping us develop appropriate management methods related to lifestyle changes. Such methods are particularly important among this population because no drug treatments are available to manage mild dyslipidemia . To the n = 534; see This study analyzed data from the Sixth Korea National Health and Nutrition Examination Survey: KNHANES VI 2015. The survey was performed in 2015 on 7380 nationally representative participants by the Korean Centers for Disease Control and Prevention (KCDCP). After written consent from the participants was obtained, trained medical and research staff conducted health examinations (BMI), self-reports , and interviews  to measure health-related variables . Data coHow many days during the past 7 days did you walk for at least 10 min at a time?’ and time they spent in light PA ‘How much time did you usually spend walking on one of those days?’. Data were represented as means (min/d).We used two items from the International Physical Activity Questionnaire (IPAQ)  to assesHow much time do you usually spend sitting or reclining on a typical day?’. Examples of SB were sitting or watching TV.We assessed this variable using items from the Korean version of the Global Physical Activity Questionnaire (GPAQ) . ParticiHow do you feel about your body shape?” [We asked participants to answer the question, “ shape?” , using a2; GL-6000-20; Cass Korea, Seoul, South Korea).To calculate BMI, we divided body weight (Seca 225) by squared height ; ability to engage in usual activities ; physical discomfort ; and anxiety/depression . Responses were weighted for conversion to an EQ-5D index based on a previously published formula [This variable was assessed using five items from the Korean version of the EuroQoL five-dimensional questionnaire (EQ-5D)  on a 3-p formula :EQ5D indWeighting was performed using the following criteria: M2 “level 2” = 1, otherwise = 0; M3 “level 3” = 1, otherwise = 0; SC2 “level 2” = 1, otherwise = 0; SC3 “level 3” = 1, otherwise = 0; UA2 “level 2” = 1, otherwise = 0; UA3 “level 3” = 1, otherwise = 0; PD2 “level 2” = 1, otherwise = 0; PD3 “level 3” = 1, otherwise = 0; AD2 “level 2” = 1, otherwise = 0; AD3 “level 3” = 1, otherwise = 0; N3 “level 3” = 1, otherwise = 0. This conversion resulted in higher scores representing better quality of life.Alcohol consumption of individuals over the past year was categorized into two groups  . Additiok) of classes was better than k-1 classes. Drinking and smoking were used as covariates across all analyses to determine latent profiles, thus controlling for other lifestyle factors. Thus, participants in the same class had homogeneous characteristics of light PA, SB, BMI, and perceived body shape. A further analysis then compared associations between latent classes and an outcome variable (auxiliary option) [d) was estimated for each group. Statistical significance was set at p < 0.05. Analyses were performed in SPSS version 22  and Mplus version 7.3 .Participants were classified according to activity level through LPA . First,  option) . Effect 2 . Overall, participants spent 452.7 min/d in SB and 38.71 min/d engaging in light PA.Participants were between 27–80 years old . Average BMI was 25.02 ± 3.35 kg/m2 see . AverageWe identified negative correlations between SB and light PA, light PA and BMI, perceived body shape and quality of life, as well as BMI and quality of life see . AdditioModels with three classes had lower Akaike Information Criterion (AIC), Bayesian Information Criterion (BIC), and sample-size adjusted BIC (SSA-BIC) values than models with one class . Models n = 48, 9%; moderate class: n = 331, 62%; inactive class, n = 154, 29%; d thresholds  [dactive-moderate = 0.80, Cohen’s dactive-inactive = 0.81), perceived body shape  and BMI  had medium effects, while SB had small effects (Cohen’s d = 0.14–0.18).We assigned names to each class based on profile characteristics (active class: ge ≥0.8) , light Pdactive-inactive = 0.49) and moderate classes (dmoderate-inactive = 0.26) had significantly higher values than the inactive class.Quality of life decreased from the active to the inactive class . Both acHere, we successfully used LPA to identify three latent classes based on SB, light PA, perceived body shape, and BMI in South Korean patients with dyslipidemia. Our results have important underlying implications for individual health. Specifically, light PA plays an important role in classifying patients with dyslipidemia. Additionally, perceived body shape is influential in increasing quality of life. Ours is the first attempt to examine whether potential contributors to quality of life among South Korean patients with dyslipidemia could be classified into latent classes. The profiles we identified provide evidence for the roles of light PA and perceived body shape in managing quality of life, knowledge that should be beneficial to healthcare practitioners.Our results suggest that health professionals should focus on promoting light PA as intervention. We found large effect sizes in light PA when discriminating between active patients and those grouped in the less active classes. Active patients engaged in roughly seven and six times more light PA than inactive and moderate patients, respectively. Indeed, light PA is the most frequently recommended form of activity  because Besides light PA, perceived body shape should be viewed as a key factor influencing quality of life. Effect sizes of perceived body shape were comparable to BMI effect sizes, and the latter is a well-established correlate of dyslipidemia ,40. IndeA limitation of this cross-sectional study is that we were unable to address any causality. Thus, we encourage further cohort and intervention studies to investigate how PA and perceived body shape alters quality of life in these contexts. Another limitation is that our study only examined patients with dyslipidemia and lacks generality. We therefore recommend caution in interpreting the results. Although numerous factors contribute to quality of life, PA and SB are widely investigated variables in research on individuals with a clinical diagnosis . In thisFor the first time, this study demonstrated that light PA, perceived body shape, and BMI are linked factors influencing quality of life. In particular, our findings indicate that more attention should be paid to perceived body shape as a contributor to quality of life in South Koreans. A priority for healthcare specialists interested in patient quality of life is to ensure the development of appropriate light-PA programs and counselling strategies to elevate body-shape perception.