Unveiling the Mainland vs. Insular Variability of the Eumerus barbarus Species Group (Diptera: Syrphidae) in the Western Mediterranean Basin.

Comprising nearly 300 described species, Eumerus Meigen, 1822, is one of the most speciose syrphid genera worldwide, and its taxonomic diversity is remarkable in the Mediterranean basin. The Eumerus barbarus (Coquebert, 1804) group consists of four species in the western Mediterranean. Although the phenotypic variability of this species group has been commented on in previous studies, it has never been contrasted with molecular data. In the present work, the morphological variation found in 300+ specimens of this species group from the western Mediterranean is explored and tested against the COI mitochondrial DNA (mtDNA). The highest phenotypic disparity was found in E. barbarus and Eumerus sulcitibius Rondani 1868. The integrative approach has not revealed cryptic diversity within the species E. barbarus but in E. sulcitibius. As a result, a new species close to E. sulcitibius was discovered, Eumerus sardus Aguado-Aranda, Ricarte & Hauser sp. n., from Sardinia, Italy. The new insular species is here described, illustrated, and discussed. A total of twenty-three haplotypes of COI mtDNA were identified amongst the analyzed Mediterranean specimens of E. barbarus, whereas two and five haplotypes were distinguished in the Iberian specimens of E. sulcitibius and Eumerus gibbosus van Steenis, Hauser & van Zuijen, 2017, respectively. Moreover, the first known barcodes of E. gibbosus and Eumerus schmideggeri van Steenis, Hauser & van Zuijen, 2017 were obtained, and the distribution ranges of all species are mapped. An updated dichotomous key to the males of the E. barbarus group from the western Mediterranean is provided.

Are Appearances Deceiving? Morpho-Genetic Complexity of the Eumerus tricolor Group (Diptera: Syrphidae) in Europe, with a Focus on the Iberian Peninsula.

Eumerus Meigen, 1822 is one of the largest Syrphidae genera in the Palaearctic Region, with the highest levels of taxonomic diversity found in the Eumerus tricolor species group. Despite its high diversity, the interspecific levels of morphological variability can be low. Additionally, some species may show certain levels of intraspecific variability. Hence, species delimitation may become challenging. In this work, we assessed the diversity of the E. tricolor group in the Iberian Peninsula through an integrative analysis of nomenclature, morphology and the 5' (COI-5') and 3' (COI-3') end regions of the Cytochrome c oxidase subunit I gene. Two new species, Eumerus ancylostylus Aguado-Aranda & Ricarte sp. n. and Eumerus petrarum Aguado-Aranda, Nedeljkovic & Ricarte sp. n., were described, and their intra- and interspecific variations discussed. In addition, the first barcodes of Iberian members of the E. tricolor group were obtained, and the distribution ranges of all species were mapped within the study area. The systematic position of the new species is discussed based on the resulting COI-based trees. The male genitalia of Eumerus hispanicus van der Goot, 1966 and Eumerus bayardi Séguy, 1961 were studied and illustrated. A lectotype was designated for Eumerus lateralis (Zetterstedt, 1819). An updated dichotomous key for all known European species of the E. tricolor group is provided. The egg of E. petrarum sp. n. is also described.

Assessing the Diversity and Systematics of Brachyopini Hoverflies (Diptera: Syrphidae) in the Iberian Peninsula, Including the Descriptions of Two New Species.

Five genera of Brachyopini, Chrysogaster Meigen, 1800, Melanogaster Rondani, 1857, Lejogaster Rondani, 1857, Orthonevra Macquart, 1829 and Riponnensia Maibach et al. 1994a are here revised from the Iberian region. Two new species, Melanogaster baetica Ricarte and Nedeljkovic, sp. n. and Orthonevra arcana Ricarte and Nedeljkovic sp. n., are described from Spain, and a third species, Chrysogaster coerulea Strobl in Czerny and Strobl, 1909 stat. n., is reinstated as valid and redescribed. A lectotype is designated for Orthonevra plumbago (Loew, 1840). The holotype of Orthonevra incisa (Loew, 1843) and the lectotype of O. plumbago are described in detail and illustrated. Melanogaster baetica sp. n. is similar to Melanogaster parumplicata (Loew, 1840) in male genitalia morphology, while O. arcana sp. n. is similar to O. incisa in the entirely-pollinose sternum I and the conspicuous incision on the posterior margin of tergum V in female. The first Iberian record of Chrysogaster rondanii Maibach and Goeldlin de Tiefenau, 1995 is provided, whilst Melanogaster aerosa is removed from the Iberian checklist of Syrphidae. Identification keys are presented to the five Brachyopini genera and 18 species now reported from the Iberian Peninsula (Chrysogaster, 6 spp.; Lejogaster, 2 spp.; Melanogaster, 3 spp.; Orthonevra, 5 spp.; Riponnensia, 2 spp.). COI (Cytochrome c oxidase subunit I) barcodes of the two new species plus C. coerulea, Chrysogaster solstitialis (Fallén, 1817), Orthonevra nobilis (Fallén, 1817) and Orthonevra frontalis (Loew, 1843) were successfully obtained from Spanish specimens. A COI-based tree was produced to locate these taxa in a wider systematic framework within the tribe.

Preferred Outcome Measures Used in Randomized Clinical Trials of Total Knee Replacement Rehabilitation: A Systematic Review.

OBJECTIVE: To determine the most frequently used outcome measures in total knee replacement rehabilitation trials. LITERATURE SURVEY: Systematic review of randomized trials searched in five databases: Web of Science, MEDical Literature Analysis and Retrieval System, Physiotherapy Evidence Database, Scopus, and Cochrane Library. METHODOLOGY: Trials were included if participants underwent total knee replacement rehabilitation and outcome measures were used to assess rehabilitation outcomes. A descriptive synthesis determined the frequency of using outcome measures and preferred assessment time points. Outcomes were classified into eight categories: patient- and clinician-reported function, performance-based function, balance, anxiety and depressive symptoms, quality of life, and others. SYNTHESIS: Eighty-one trials were included and 102 different outcome measures were classified. The most frequently reported outcome was knee range of motion, used in 54% of trials, followed by a visual analog scale of pain (43%) and Western Ontario and McMaster Universities Arthritis Index (WOMAC; 40%). Patient- and clinician-reported function were the categories most frequently assessed (74%), whereas performance-based measures were implemented by 56% of trials. The most frequent assessment time points were 1 week presurgery (52%) and 3 months postsurgery (39%). CONCLUSIONS: There is consensus regarding the need to evaluate functional outcomes in total knee replacement rehabilitation trials but none regarding the outcome measure that should be used. These findings suggest that most trials include patient- and clinician-reported functional measures, along with pain and performance-based measures in trial designs.

The study of hidden habitats sheds light on poorly known taxa: spiders of the Mesovoid Shallow Substratum.

The scarce and biased knowledge about the diversity and distribution of Araneae species in the Iberian Peninsula is accentuated in poorly known habitats such as the Mesovoid Shallow Substratum (MSS). The aim of this study was to characterize the spiders inventory of the colluvial MSS of the Sierra de Guadarrama National Park, and to assess the importance of this habitat for the conservation of the taxon. Thirty-three localities were selected across the high peaks of the Guadarrama mountain range and they were sampled for a year using subterranean traps specially designed to capture arthropods in the MSS. Species accumulation curves were built both for the observed species richness and for the non-parametric richness estimators. The literature was reviewed in order to update the distributional maps of the rarest species. Forty-two species were collected, of which four were species new to science. More than half were represented by one or two individuals which caused the accumulation curves to rise slowly and to end without reaching an asymptote. Almost half of the species showed significant increases in their Iberian distribution ranges. Two species were recorded for the first time in the Iberian Peninsula and 32 species were new additions to the spider checklist of the Sierra de Guadarrama National Park.

Adipose tissue-derived mesenchymal stem cells: isolation, expansion, and characterization.

Over the last decade, cell therapy has emerged as a potentially new approach for the treatment of cardiovascular diseases. Among the wide range of cell types and sources, adipose-derived mesenchymal stem cells have shown promise, mainly due to its plasticity and remarkable paracrine-secretion capacity, largely demonstrated at the in vitro and in vivo levels. Furthermore, its accessibility and abundance, the low morbidity of the surgical procedure, its easy isolation, culture, and long-term passaging capacity added to its immunomodulatory properties that could allow its allogeneic transplantation, making it one of the most attractive candidates for clinical application. In this chapter, we will focus on the methodology for the isolation, expansion, phenotypical characterization, differentiation, and storage of the adipose-derived stem cells.

Culture of human bone marrow-derived mesenchymal stem cells on of poly(L-lactic acid) scaffolds: potential application for the tissue engineering of cartilage.

PURPOSE: Due to the attractive properties of poly(L-lactic acid) (PLLA) for tissue engineering, the aim was to determine the growth and differentiation capacity of mesenchymal stromal cells (MSCs) in PLLA scaffolds and their potential use in the treatment of cartilage diseases. METHODS: MSCs were cultured in PLLA films and thin porous membranes to study adherence and proliferation. Permeability and porosity were determined for the different scaffolds employed. The optimal conditions for cell seeding were first determined, as well as cell density and distribution inside the PLLA. Scaffolds were then maintained in expansion or chondrogenic differentiation media for 21 days. Apoptosis, proliferation and chondrogenic differentiation was assessed after 21 days in culture by immunohistochemistry. Mechanical characteristics of scaffolds were determined before and after cell seeding. RESULTS: MSCs uniformly adhered to PLLA films as well as to porous membranes. Proliferation was detected only in monolayers of pure PLLA, but was no longer detected after 10 days. Mechanical characterization of PLLA scaffolds showed differences in the apparent compression elastic modulus for the two sizes used. After determining high efficiencies of seeding, the production of extracellular matrix (ECM) was determined and contained aggrecan and collagens type I and X. ECM produced by the cells induced a twofold increase in the apparent elastic modulus of the composite. CONCLUSIONS: Biocompatible PLLA scaffolds have been developed that can be efficiently loaded with MSCs. The scaffold supports chondrogenic differentiation and ECM deposition that improves the mechanics of the scaffold. Although this improvement does not met the expectations of a hyaline-like cartilage ECM, in part due to the lack of a mechanical stimulation, their potential use in the treatment of cartilage pathologies encourages to improve the mechanical component.

Transplantation of mesenchymal stem cells exerts a greater long-term effect than bone marrow mononuclear cells in a chronic myocardial infarction model in rat.

The aim of this study is to assess the long-term effect of mesenchymal stem cells (MSC) transplantation in a rat model of chronic myocardial infarction (MI) in comparison with the effect of bone marrow mononuclear cells (BM-MNC) transplant. Five weeks after induction of MI, rats were allocated to receive intramyocardial injection of 10(6) GFP-expressing cells (BM-MNC or MSC) or medium as control. Heart function (echocardiography and (18)F-FDG-microPET) and histological studies were performed 3 months after transplantation and cell fate was analyzed along the experiment (1 and 2 weeks and 1 and 3 months). The main findings of this study were that both BM-derived populations, BM-MNC and MSC, induced a long-lasting (3 months) improvement in LVEF (BM-MNC: 26.61 +/- 2.01% to 46.61 +/- 3.7%, p < 0.05; MSC: 27.5 +/- 1.28% to 38.8 +/- 3.2%, p < 0.05) but remarkably, only MSC improved tissue metabolism quantified by (18)F-FDG uptake (71.15 +/- 1.27 to 76.31 +/- 1.11, p < 0.01), which was thereby associated with a smaller infarct size and scar collagen content and also with a higher revascularization degree. Altogether, results show that MSC provides a long-term superior benefit than whole BM-MNC transplantation in a rat model of chronic MI.

Effects of birth weight on anti-mullerian hormone serum concentrations in infant girls.

CONTEXT: We previously demonstrated that low birth weight (BW) infant girls show increased serum anti-Müllerian hormone (AMH) concentrations and poststimulated estradiol levels compared to normal-BW infants, suggesting an altered follicular development. However, the impact of high BW on reproductive function is less known. OBJECTIVE: To evaluate the effect of BW on AMH, we determined the concentrations of this hormone in low-BW, normal-BW, and high-BW female infants during the first 3 months of life. DESIGN: Twenty-seven low-BW, 29 normal-BW, and 28 high-BW infant girls were studied. We measured serum gonadotropins, steroid hormones, AMH, glucose, insulin, free fatty acids, IGF-I, and adiponectin in a fasting blood sample. In addition, in a subgroup of normal-BW (n = 23) and high-BW infants (n = 10), a GnRH analog leuprolide acetate test was performed. RESULTS: Serum concentrations of AMH were higher in low-BW and high-BW infants compared to normal-BW infants (P = 0.028 and 0.022, respectively). In addition, in high-BW infants, adiponectin concentrations were lower (P = 0.018), and poststimulated FSH and estradiol levels were higher compared to normal-BW infants (P = 0.024 and 0.047, respectively). CONCLUSIONS: Serum AMH and poststimulated estradiol concentrations are increased in low-BW and high-BW female infants, suggesting that these girls may show evidence of an altered follicular development. However, the increased poststimulated FSH levels and low adiponectin concentrations observed in high-BW infants suggest that ovarian function is perturbed through a different mechanism from that in low-BW infants.

Epigenetic signatures associated with different levels of differentiation potential in human stem cells.

BACKGROUND: The therapeutic use of multipotent stem cells depends on their differentiation potential, which has been shown to be variable for different populations. These differences are likely to be the result of key changes in their epigenetic profiles. METHODOLOGY/PRINCIPAL FINDINGS: to address this issue, we have investigated the levels of epigenetic regulation in well characterized populations of pluripotent embryonic stem cells (ESC) and multipotent adult stem cells (ASC) at the trancriptome, methylome, histone modification and microRNA levels. Differences in gene expression profiles allowed classification of stem cells into three separate populations including ESC, multipotent adult progenitor cells (MAPC) and mesenchymal stromal cells (MSC). The analysis of the PcG repressive marks, histone modifications and gene promoter methylation of differentiation and pluripotency genes demonstrated that stem cell populations with a wider differentiation potential (ESC and MAPC) showed stronger representation of epigenetic repressive marks in differentiation genes and that this epigenetic signature was progressively lost with restriction of stem cell potential. Our analysis of microRNA established specific microRNA signatures suggesting specific microRNAs involved in regulation of pluripotent and differentiation genes. CONCLUSIONS/SIGNIFICANCE: Our study leads us to propose a model where the level of epigenetic regulation, as a combination of DNA methylation and histone modification marks, at differentiation genes defines degrees of differentiation potential from progenitor and multipotent stem cells to pluripotent stem cells.

Comparison of ex vivo expansion culture conditions of mesenchymal stem cells for human cell therapy.

BACKGROUND: Mesenchymal stem cells (MSCs) are multipotent stem cells. Based on their properties, several clinical trials have been designed to explore their potential therapeutic effect. Fetal calf serum (FCS, commonly used for in vitro expansion) is an undesirable source of xenogeneic antigens and bears the risk of transmitting contaminations. As an alternative for FCS, platelet lysate (PL) and both autologous and allogeneic human serum have been proposed. The aim of this study is to compare the culture of bone marrow (BM)-derived MSCs in the presence of different serum supplements to determine the effect on cell growth, differentiation potential, and immunologic function. STUDY DESIGN AND METHODS: MSCs from BM of healthy volunteer donors were grown in the presence of 10% FCS supplemented with 1 ng/mL basic fibroblast growth factor (bFGF), 10% human serum supplemented with 1 ng/mL bFGF, 5% PL, and PL 5% supplemented with 1 ng/mL bFGF (PL plus bFGF). RESULTS: MSCs that expanded in either medium showed a comparable morphology, phenotype, and proliferative and differentiation capacity. While the presence of MSCs in vitro significantly decreased CD3/CD28-mediated T-cell activation, this effect was significantly higher in MSCs cultured with human serum. Production of interferon-gamma was inhibited by cocultured media with MSCs while MSCs also induced a significant inhibition of cell cycle in T cells. DISCUSSION: In conclusion, PL or autologous serum could offer an alternative to the use of FCS in MSC expansion for clinical use maintaining the same growing potential, phenotype, immunomodulatory properties, and differentiation potential.

Metabolic parameters in cord blood of newborns of women with polycystic ovary syndrome.

OBJECTIVE: To assess metabolic parameters in the cord blood of newborns of women with polycystic ovary syndrome (PCOS) and to correlate these parameters with those of mothers with PCOS during midgestation. DESIGN: Case-control study. SETTING: Unit of Endocrinology and Reproductive Medicine. PATIENT(S): Thirty newborns of mothers with PCOS (PCOSn) and 34 newborns of control mothers (Cn) were studied. INTERVENTION(S): A sample of cord blood was obtained at delivery. In all mothers, an oral glucose tolerance test (oGTT) with measurement of glucose and insulin was performed at 22-28 weeks of gestation. In cord blood and in the fasting sample of the oGTT, serum leptin, adiponectin, insulin, glucose, and lipids (triglycerides, cholesterol, high-density lipoprotein cholesterol, and low-density lipoprotein cholesterol) were determined. RESULT(S): PCOSn showed significantly higher leptin concentrations than Cn. Moreover, in PCOSn, leptin concentrations in cord blood were correlated with birth weight (r = 0.495) and body mass index of the mother at midpregnancy (r = 0.644). CONCLUSION(S): The metabolic parameters in the cord blood of PCOSn are similar to those observed in controls, except for leptin concentrations, which are significantly higher. The latter could be related to the fetal adiposity or the metabolic condition of the mother.

Characterization of the paracrine effects of human skeletal myoblasts transplanted in infarcted myocardium.

BACKGROUND: The discrepancy between the functional improvements yielded experimentally by skeletal myoblasts (SM) transplanted in infarcted myocardium and the paucity of their long-term engraftment has raised the hypothesis of cell-mediated paracrine mechanisms. METHODS AND RESULTS: We analyzed gene expression and growth factors released by undifferentiated human SM (CD56(+)), myotubes (SM cultured until confluence) and fibroblasts-like cells (CD56(-)). Gene expression revealed up-regulation of pro-angiogenic (PGF), anti-apoptotics (BAG-1, BCL-2), heart development (TNNT2, TNNC1) and extracellular matrix remodelling (MMP-2, MMP-7) genes in SM. In line with the gene expression profile, the analysis of culture supernatants of SM by ELISA identified the release of growth factors involved in angiogenesis (VEGF, PIGF, angiogenin, angiopoietin, HGF and PDGF-BB) as well as proteases involved in matrix remodelling (MMP2, MMP9 and MMP10) and their inhibitors (TIMPs). Culture of smooth muscle cells (SMC), cardiomyocytes (HL-1) and human umbilical vein endothelial cells (HUVECs) with SM-released conditioned media demonstrated an increased proliferation of HUVEC, SMC and cardiomyocytes (p<0.05) and a decrease in apoptosis of cardiomyocytes (p<0.05). Analysis of nude rats transplanted with human SM demonstrated expression of human-specific MMP-2, TNNI3, CNN3, PGF, TNNT2, PAX7, TGF-beta, and IGF-1 1 month after transplant. CONCLUSIONS: Our data support the paracrine hypothesis whereby myoblast-secreted factors may contribute to the beneficial effects of myogenic cell transplantation in infarcted myocardium.