A high-density consensus map of A and B wheat genomes

A durum wheat consensus linkage map was developed by combining segregation data from six mapping populations. All of the crosses were derived from durum wheat cultivars, except for one accession of T. ssp. dicoccoides. The consensus map was composed of 1,898 loci arranged into 27 linkage groups covering all 14 chromosomes. The length of the integrated map and the average marker distance were 3,058.6 and 1.6 cM, respectively. The order of the loci was generally in agreement with respect to the individual maps and with previously published maps. When the consensus map was aligned to the deletion bin map, 493 markers were assigned to specific bins. Segregation distortion was found across many durum wheat chromosomes, with a higher frequency for the B genome. This high-density consensus map allowed the scanning of the genome for chromosomal rearrangements occurring during the wheat evolution. Translocations and inversions that were already known in literature were confirmed, and new putative rearrangements are proposed. The consensus map herein described provides a more complete coverage of the durum wheat genome compared with previously developed maps. It also represents a step forward in durum wheat genomics and an essential tool for further research and studies on evolution of the wheat genome. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s00122-012-1939-y) contains supplementary material, which is available to authorized users

In vitro analysis of the effects on wound healing of high- and low-molecular weight chains of hyaluronan and their hybrid H-HA/L-HA complexes

Abstract Background: Recent studies have reported the roles of Hyaluronic acid (HA) chains of diverse length in wound repair, especially considering the simultaneous occurrence in vivo of both high- (H-HA) and low-molecular weight (L-HA) hyaluronan at an injury site. It has been shown that HA fragments (5 ≤ MW ≤ 20 kDa) usually trigger an inflammatory response that, on one hand, is the first signal in the activation of a repair mechanism but on the other, when it’s overexpressed, it may promote unwanted side effects. The present experimental research has aimed to investigate H-HA, L-HA and of a newly developed complex of the two (H-HA/L-HA) for stability (e.g. hyaluronidases digestion), for their ability to promote wound healing of human keratinocytes in vitro and for their effect on cellular biomarker expression trends. Results: Time-lapse video microscopy studies proved that the diverse HA was capable of restoring the monolayer integrity of HaCat. The H-HA/L-HA complex (0.1 and 1%w/v) proved faster in regeneration also in co-culture scratch test where wound closure was achieved in half the time of H-HA stimulated cells and 2.5-fold faster than the control. Gene expression was evaluated for transformation growth factor beta 1 (TGF-β1) proving that L-HA alone increased its expression at 4 h followed by restoration of similar trends for all the stimuli. Depending on the diverse stimulation (H-HA, L-HA or the complex), metalloproteinases (MMP-2, -9, -13) were also modulated differently. Furthermore, type I collagen expression and production were evaluated. Compared to the others, persistence of a significant higher expression level at 24 h for the H-HA/L-HA complex was found. Conclusions: The outcomes of this research showed that, both at high and low concentrations, hybrid complexes proved to perform better than HA alone thus suggesting their potential as medical devices in aesthetic and regenerative medicine. Keywords: Wound healing, Hyaluronan, MMPs, Hybrid complexe

Mortality and pulmonary complications in patients undergoing surgery with perioperative SARS-CoV-2 infection: an international cohort study

Background: The impact of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) on postoperative recovery needs to be understood to inform clinical decision making during and after the COVID-19 pandemic. This study reports 30-day mortality and pulmonary complication rates in patients with perioperative SARS-CoV-2 infection. Methods: This international, multicentre, cohort study at 235 hospitals in 24 countries included all patients undergoing surgery who had SARS-CoV-2 infection confirmed within 7 days before or 30 days after surgery. The primary outcome measure was 30-day postoperative mortality and was assessed in all enrolled patients. The main secondary outcome measure was pulmonary complications, defined as pneumonia, acute respiratory distress syndrome, or unexpected postoperative ventilation. Findings: This analysis includes 1128 patients who had surgery between Jan 1 and March 31, 2020, of whom 835 (74·0%) had emergency surgery and 280 (24·8%) had elective surgery. SARS-CoV-2 infection was confirmed preoperatively in 294 (26·1%) patients. 30-day mortality was 23·8% (268 of 1128). Pulmonary complications occurred in 577 (51·2%) of 1128 patients; 30-day mortality in these patients was 38·0% (219 of 577), accounting for 81·7% (219 of 268) of all deaths. In adjusted analyses, 30-day mortality was associated with male sex (odds ratio 1·75 [95% CI 1·28–2·40], p\textless0·0001), age 70 years or older versus younger than 70 years (2·30 [1·65–3·22], p\textless0·0001), American Society of Anesthesiologists grades 3–5 versus grades 1–2 (2·35 [1·57–3·53], p\textless0·0001), malignant versus benign or obstetric diagnosis (1·55 [1·01–2·39], p=0·046), emergency versus elective surgery (1·67 [1·06–2·63], p=0·026), and major versus minor surgery (1·52 [1·01–2·31], p=0·047). Interpretation: Postoperative pulmonary complications occur in half of patients with perioperative SARS-CoV-2 infection and are associated with high mortality. Thresholds for surgery during the COVID-19 pandemic should be higher than during normal practice, particularly in men aged 70 years and older. Consideration should be given for postponing non-urgent procedures and promoting non-operative treatment to delay or avoid the need for surgery. Funding: National Institute for Health Research (NIHR), Association of Coloproctology of Great Britain and Ireland, Bowel and Cancer Research, Bowel Disease Research Foundation, Association of Upper Gastrointestinal Surgeons, British Association of Surgical Oncology, British Gynaecological Cancer Society, European Society of Coloproctology, NIHR Academy, Sarcoma UK, Vascular Society for Great Britain and Ireland, and Yorkshire Cancer Research

Mortality from gastrointestinal congenital anomalies at 264 hospitals in 74 low-income, middle-income, and high-income countries: a multicentre, international, prospective cohort study

Summary Background Congenital anomalies are the fifth leading cause of mortality in children younger than 5 years globally. Many gastrointestinal congenital anomalies are fatal without timely access to neonatal surgical care, but few studies have been done on these conditions in low-income and middle-income countries (LMICs). We compared outcomes of the seven most common gastrointestinal congenital anomalies in low-income, middle-income, and high-income countries globally, and identified factors associated with mortality. Methods We did a multicentre, international prospective cohort study of patients younger than 16 years, presenting to hospital for the first time with oesophageal atresia, congenital diaphragmatic hernia, intestinal atresia, gastroschisis, exomphalos, anorectal malformation, and Hirschsprung’s disease. Recruitment was of consecutive patients for a minimum of 1 month between October, 2018, and April, 2019. We collected data on patient demographics, clinical status, interventions, and outcomes using the REDCap platform. Patients were followed up for 30 days after primary intervention, or 30 days after admission if they did not receive an intervention. The primary outcome was all-cause, in-hospital mortality for all conditions combined and each condition individually, stratified by country income status. We did a complete case analysis. Findings We included 3849 patients with 3975 study conditions (560 with oesophageal atresia, 448 with congenital diaphragmatic hernia, 681 with intestinal atresia, 453 with gastroschisis, 325 with exomphalos, 991 with anorectal malformation, and 517 with Hirschsprung’s disease) from 264 hospitals (89 in high-income countries, 166 in middleincome countries, and nine in low-income countries) in 74 countries. Of the 3849 patients, 2231 (58·0%) were male. Median gestational age at birth was 38 weeks (IQR 36–39) and median bodyweight at presentation was 2·8 kg (2·3–3·3). Mortality among all patients was 37 (39·8%) of 93 in low-income countries, 583 (20·4%) of 2860 in middle-income countries, and 50 (5·6%) of 896 in high-income countries (p<0·0001 between all country income groups). Gastroschisis had the greatest difference in mortality between country income strata (nine [90·0%] of ten in lowincome countries, 97 [31·9%] of 304 in middle-income countries, and two [1·4%] of 139 in high-income countries; p≤0·0001 between all country income groups). Factors significantly associated with higher mortality for all patients combined included country income status (low-income vs high-income countries, risk ratio 2·78 [95% CI 1·88–4·11], p<0·0001; middle-income vs high-income countries, 2·11 [1·59–2·79], p<0·0001), sepsis at presentation (1·20 [1·04–1·40], p=0·016), higher American Society of Anesthesiologists (ASA) score at primary intervention (ASA 4–5 vs ASA 1–2, 1·82 [1·40–2·35], p<0·0001; ASA 3 vs ASA 1–2, 1·58, [1·30–1·92], p<0·0001]), surgical safety checklist not used (1·39 [1·02–1·90], p=0·035), and ventilation or parenteral nutrition unavailable when needed (ventilation 1·96, [1·41–2·71], p=0·0001; parenteral nutrition 1·35, [1·05–1·74], p=0·018). Administration of parenteral nutrition (0·61, [0·47–0·79], p=0·0002) and use of a peripherally inserted central catheter (0·65 [0·50–0·86], p=0·0024) or percutaneous central line (0·69 [0·48–1·00], p=0·049) were associated with lower mortality. Interpretation Unacceptable differences in mortality exist for gastrointestinal congenital anomalies between lowincome, middle-income, and high-income countries. Improving access to quality neonatal surgical care in LMICs will be vital to achieve Sustainable Development Goal 3.2 of ending preventable deaths in neonates and children younger than 5 years by 2030

Discoid Nanoparticles: pH-dependent Size Response

Introduction: Tumor targeting is a major issue point in development of new drugs. The targeting can be making by conjugating specific targeting molecole or modifying their size or shape. Discoidal Nanopartcles (DNs) have discoidal shape and are very small. DNs is obtained from liposomes by adding Styrene-Maleic Acid copolimer (SMAnh) (Fig. 1). Molecular ratio between styrene and maleic anhydride and pH of microenvironment reaction can however affect the synthesis of DNs [1]. In this work we studied the use of SMAnh as copolymers capable to synthesis DNs starting from spherical liposomes at different molar ratio of copolymer and pH. Materials and Methods: Different liposomes formulations were synthesized using thin layer evaporation method, freeze and thaw and extrusion. The DNs are synthesized by incubating the best resulting liposomes (DMPC) with SMAnh with molecular ratio 2:1 in Hepes 10 mM at pH=7.4; pH=6.5 and pH=5.5. The liposomes and resulting DNs are further physiochemically characterized by DLS and UV-Vis spectrophotometer (Fig. 2). Results and Discussion: The SMAnh copolymer is able to form DNs, but its behavior depends on pH enviroment. Infact, at several pH is possible find different particle sizes. Currently the best enviroment condition is at pH=6.5. Conclusion: DNs could be a innovative drug delivery systems for anticancer therapy. SMAnh molar ratio affects the synthesis of DNs as well as drug delivery

Soil aggregates, organic matter turnover and carbon balance in a Mediterranean eroded vineyard

The carbon cycle is being affected by the human impacts (Novara et al., 2011; Yan-Gui et al., 2013), and one of those is the intensification in the soil erosion in agriculture land (Cerdà et al., 2009; García Orenes et al., 2009). Vineyards also are affected by the human activities (Fernández Calviño, 2012). Vineyards in Sicily are cultivated on 110.000 ha, 10% of which on >10% slope. Deficiencies of soil organic matter are typical of the semi arid Mediterranean environment especially where traditional intensive cropping practices are adopted (Novara et al., 2012; 2013). These practices in vineyards could lead soil to intensive erosion processes (Novara et al., 2011). The fate of SOC under erosion processes is difficult to understand because of the influence of the erosion impact on SOC pathway, which depends on the different features of the process involved (detachment, transport and/or deposition). Soil erosion must be considered a net C source (Lal, 2003), as eroded soils have lower net primary productivity (NPP) (Dick and Gregorich, 2004) caused by reduction in the effective rooting depth and all in all determining decline in soil quality. Breakdown of aggregates and soil dispersion expose SOM to microbial/enzymatic processes and chemical soil properties (Dimoyiannis, 2012; Kocyigit and Demirci, 2012). Moreover the light fraction, transported by runoff, is labile and easily mineralized determining CO2 emission in the atmosphere (Jacinthe and Lal, 2004). Therefore, the carbon pool is lower in eroded than in un-eroded soil scapes and the rate of mineralization of soil organic matter is higher in sediments than in original soil. In this survey we show a research conducted on a slope sequence of three soil profiles in an irrigated vineyard located in Sambuca di Sicilia, Italy (UTM33-WGS84: 4169367N; 325011E). The SOC content was measured at depth intervals of 10 cm up to a depth of 60 cm in each pedon.Wet aggregate-size fractions with no prior chemical dispersion, were isolated by mechanical shaking of 100 g, air-dried fine earth on a column with sieves of 250 and 63 m using a Shaker AS 200 Sieve (RETSCH analytical, Haan, Germany) (200-mm sieves, amplitude of 2 cm, frequency of 1.6 Hz and a water flux of 2 litres minute-1). After the physical fractionation, we discriminate three main aggregate-size fractions: >250, 63–250 and <63 m. Three replicate samples of 5 g of the soil material that we prepared for the fractionation from three different pedons along the slope gradient were incubated at two different depth intervals (Topsoil: 0–15 cm; Subsoil: 35–50 cm). Respiration was monitored during a period of 50 days keeping moisture and temperature constant. Both in topsoil and subsoil layers, particle size distribution in the depositional area shows a decrease of the finest size (<63 m) respect to the soil in the detachment area. A SOC increase was observed due to depositional processes. Mean Residence Time of SOC strongly decreased in the subsoil particularly in the depositional area corroborating that erosion processes could be a SOC sink. Anyway we should also stress that, considering the estimated “off farm” erosion processes, the carbon budget resulted highly negative

Development of hybrid nanocarrier library based biomaterials for cancer therapy

Introduction: The major challenges are requested to improve the properties of nanoparticles injected systemically: i) the increase of payloads inside nanoparticles, ii) the controlled release of payloads, iii) the decrease of interaction and uptake with mononuclear phagocyte system cells, iv) the selective targeting. Geometry and size of the therapeutic nanoparticles play a crucial role for their adhesion to tumor vasculature1. Discoidal non-spherical nanoparticles interact better with the fenestrated endothelium of tumor tissue with respect to spherical nanoparticles. Recently, discoidal nanoparticles have been developed to delivery payloads for anticancer therapy as well as cardiovascular diseases2. The aim of this project is of the synthesis of discoidal and hybrid nanocarrier (DHN) obtained by combining biomaterials, e.g. lipid/polymer, and extracellular vesicles for the treatment of Hepatoma and related-diseases. In particular, DHN shall allow to cure the liver diseases (i.e., cancer) and to stimulate the regeneration of tissue. Materials and Methods: DHN will be designed, synthesized and physicochemical characterize during different techniques such as DLS, NTA,TEM, FT-IR, LC ms/ms, and HPLC. In vitro and in vivo experiment will be carried out to test the toxicity and pre-clinic efficacy in suitable hepatic cellular lines and murine models. Conclusion: There resulting DHN may represent a new nanomedicine for the treatment of Hepatoma and related-diseases

Discoidal Nanoparticles: reaction environment-dependent Size Response

In recent years, the develop of new drug delivery system (DDS) was necessary to optimize the treatment efficiency, thus overcoming the limits of traditional teraphy such as targeting and drug half life. However, many treatmants cause immunogenicity. Several DDS was developed such as liposomes, niosomes and other tipe of nanoparticles, but they can have some limits like the recognition and elimination of the immune system or the tumour targeting. These drawbacks can be bypassed by modifing sizes and shapes of liposomes. Discoidal Nanopartcles (DNs) are obtained from liposomes by adding Styrene-Maleic Acid copolimer (SMA) (Fig. 1). The molecular ratio between styrene and maleic anhydride, pH and temperature of microenvironment reaction can affect the synthesis of DNs [1]. In this work we studied the use of SMA as copolymer, which is able to synthesis DNs by starting from spherical liposomes (DMPC) at different molar ratio of copolymers (2:1 and 4:1), pHs (range of pH from 3.5 to 11.5) and temperatures (4°C, 25°C, 37°C, 65°C). The SMA copolymer can form DNs, and their properties depend on the reaction enviroment. In fact, at different pHs, particle sizes are modified according to these physical parameters. The modifcation of temperature can influence the synthesis of DNs. Currently, the best condition is obtained at pH = 7.4 and 25°C by using 2:1 molar ratio of copolymer. The results are in agreement with previously reported data using SMA as a copolymer under different reaction conditions [1]. These properties could be affected the synthesis of DNs as well as drug delivery, but DNs could be a innovative DDS for anticancer therapy. Acknowledgment: This research is funded by the Italian Ministry of Instruction, University, and Research under the national project PON Ricerca e Innovazione 2014-2020

HYBRID NANOCARRIER LIBRARY BASED BIOMATERIALS FOR THERAPEUTIC APPLICATIONS.

Tumor targeting plays a pivot role in anticancer therapy. Tumor targeting can be carried out by conjugating specific targeting agents on the surface of nanoparticles or modifying their shape and size. To improve the selective targeting and accumulate therapeutic agents inside the tumor tissues, we designed new Discoidal Nanoparticles (DNs) having a non-spherical shape that better interact and cross through the fenestrated endothelium of tumor tissue compared to spherical nanoparticles. These nanoparticles are lipid based and are made up from the same lipids used to make liposomes and are obtained by changing the original spherical shape of liposomes in discoidal and decreasing liposomal native average sizes. Copolymers are basically used to modify spherical shape of liposomes and finally obtain discoidal form. SMAnh is a copolymer synthesized by reversible addition-fragmentation chain transfer polymerization (RAFT) between Styrene and Maleic Anhydride and can form nanodisks. Molecular ratio between styrene and maleic anhydride, pH and temperature of microenvironment reaction can however affect the synthesis of nanodiscks [1]. In this work we studied the use of SMAnh as copolymers capable to synthesis DNs starting from spherical liposomes at different molar ratio of copolymer, pH and temperature. MATERIALS AND METHODS Different liposomes were synthesized using thin layer evaporation method, freeze and thaw and extrusion. Liposomes were physicochemical characterized by using Dinamic Light Scattering (DLS) and UV-Vis spectrophotometer. The best selected formulations was DMPC liposomes that was used for further experiments. The DNs are synthesized by incubating the resulting liposomes with SMAnh (Styrene-Anhydride maleic molecular ratio 2:1) in Hepes 10 mM at pH = 7.4 and pH = 6.4. The resulting DNs are further physiochemically characterized by DLS and UV-Vis spectrophotometer. RESULT AND DISCUSSION SMAnh 2:1 molar ratio cannot make only DN at pH = 7.4. UV-Vis absorption of DN increased over time. This effect is directly related to the turbidity of nanoparticles. UV-Vis analysis agreed DLS data that show the presence of three different particle sizes at 5000 nm (80%), 200 nm (13%) and 13 nm (7%). Large particles depend on the presence of free SMAnh making aggregates in incubation medium. Narrow size distributed DN was obtained at pH = 6.4 using SMAnh 2:1 (molar ratio). DLS data shows two different peaks: 200 nm (10%) that represents bare liposomes and 13 nm (60%) that represents DN (Figure 1)

Properties of Newly-Synthesized Cationic Semi-Interpenetrating Hydrogels Containing Either Hyaluronan or Chondroitin Sulfate in a Methacrylic Matrix

Extracellular matrix components such as hyaluronan (HA) and chondroitin sulfate (CS) were combined with a synthetic matrix of p(HEMA-co-METAC) (poly(2-hydroxyethylmethacrylate-co-2-methacryloxyethyltrimethylammonium)) at 1% and 2% w/w concentration following a previously developed procedure. The resulting semi-interpenetrating hydrogels were able to extensively swell in water incrementing their dry weight up to 13 fold depending on the glycosamminoglycan content and nature. When swollen in physiological solution, materials water uptake significantly decreased, and the differences in swelling capability became negligible. In physiological conditions, HA was released from the materials up to 38%w/w while CS was found almost fully retained. Materials were not cytotoxic and a biological evaluation, performed using 3T3 fibroblasts and an original time lapse videomicroscopy station, revealed their appropriateness for cell adhesion and proliferation. Slight differences observed in the morphology of adherent cells suggested a better performance of CS containing hydrogels