The massive use of synthetic plastics, in particular in the food packaging area, has a great environmental impact, and alternative more ecologic materials are being required. Poly(lactic) acid (PLA) and starch have been extensively studied as potential replacements for non-degradable petrochemical polymers on the basis of their availability, adequate food contact properties and competitive cost. Nevertheless, both polymers exhibit some drawbacks for packaging uses and need to be adapted to the food packaging requirements. Starch, in particular, is very water sensitive and its film properties are heavily dependent on the moisture content, exhibiting relatively low mechanical resistance. PLA films are very brittle and offer low resistance to oxygen permeation. Their combination as blend or multilayer films could provide properties that are more adequate for packaging purposes on the basis of their complementary characteristics. The main characteristics of PLA and starch in terms of not only the barrier and mechanical properties of their films but also of their combinations, by using blending or multilayer strategies, have been analyzed, identifying components or processes that favor the polymer compatibility and the good performance of the combined materials. The properties of some blends/combinations have been discussed in comparison with those of pure polymer films. Over the last few decades, the environment has become one of the major global concerns, especially in the face of pollution, the depletion of natural resources and environmental degradation. As a consequence, research efforts have been developed at different levels to find alternative solutions. A specific concern is the field of packaging, which produces great amounts of non-degradable plastic waste accumulated in critical areas around the planet, causing severe problems and representing high recycling costs. Europe ranks second, along with North America, in the global production of plastic materials with 18.5% of the worldwide annual production . In Europe, the packaging sector is the largest one in the plastics industry, since it represents 40% of the total plastic demand, which rises to 49 million tons  at three different contents  were produced by extrusion method [Another option considered for the purposes of improving PLA properties is to combine it with other biopolymers by using an adequate compatibilization strategy. Different PLA blends have been studied, such as PLA-PHB/cellulose nanocrystals (CNCs) plasticized with 15 wt. % of acetyl(tributylcitrate) (ATBC) . The inct et al. . From tht et al.  also devn method . The stuStarch is a plant polysaccharide consisting of several glucose units joined by glycosidic bonds. It is found in cereals (30–70% of the dry matter), tubers (60–90%) and legumes (25–50%) . It is o−13 and 2.5 × 10−13 cm3·m−1·s−1·Pa−1, depending on the film formulation and film-forming process [−7 and 8.3 × 10−5 g·m−1·s−1·Pa−1, depending on the film plasticization level or moisture content [Starch-based films have demonstrated several advantages, such as their extensibility and good oxygen barrier properties ,56. The  process ,56,57. T process . Moreove process . However content ,61. It i content ,63. TablStarch-based films can be obtained by both thermo processing and casting techniques ,68,69,70Many applications have been described for starch. Indeed, it can be converted into chemicals  and used in the production of synthetic polymers, it can be transformed into other biopolymers through fermentative processes or it can be hydrolyzed to provide monomers or oligomers. Likewise, it can be grafted with a variety of reagents to produce new derivative polymeric materials for different uses . Starch Thermoplastic starch, which is a rubbery material, is obtained by adding plasticizers to the native starch during its processing. Plasticizers can be fructose , sorbitoTo remedy the previously described disadvantages of both PLA and starch and to reduce the cost of the finished products, some alternatives have been figured out. Since both polymers exhibit opposite mechanical and barrier properties, their combination could lead to films with improved functional properties. However, since starch is highly hydrophilic and PLA is hydrophobic, they are thermodynamically immiscible. The biggest issue with mixing both polymers relies on the phase separation. To enhance their compatibility, either starch  or PLA  can be mSeveral studies were also carried out using maleic anhydride, grafted either on PLA ,88 and/oLe Bolay et al.  investigAs previously reported, the map of tensile properties of neat PLA and starch a, shows Although relevant results showed that PLA-starch blending seems to be a good alternative method to enhance the film properties of net PLA or starch films, this requires the involvement of reactive processes, which, in turn, could lead to reactant residues in the films which must be tested as to their food safety. In contrast, the strategy of the bi or multilayer films did not require such great efforts to enhance the interfacial properties of the polymers since there is only contact at the layer interface. Moreover, in terms of barrier properties, a multilayer assembly exhibits more advantages, since the complementary barrier capacity of each polymer can yield a composite material with extremely high resistance to the mass transport of polar (water) and non-polar (gases) molecules, resulting in a very effective packaging material. Nevertheless, the good interfacial adhesion of the materials where there is contact between the layers must be ensured from the mechanical point of view. This aspect could be enhanced by using adequate adhesive materials, but studies on their migration are necessary, especially for food applications, where food safety can be compromised. Some studies carried out into biopolymer bilayer films are summarized in the next section.Very few multilayer film strategies have been studied, with the objective to optimize the functional properties of biodegradable packaging materials. Escherichia coli and Listeria innocua. Although tensile properties were not improved with respect to pure PHBV films, the active compounds yielded more transparent materials with improved WVP. The preparation and characterization of biodegradable bilayer films from isolated soy protein (SPI) and PLA were reported by González and Alvarez Igarzabal [Starch-PCL bilayer films were also obtained by Ortega et al.  by comprgarzabal . The tengarzabal , obtainigarzabal , obtaineThe multilayer strategy is often used in the packaging industry and more and more of the food available in the stores comes in high-tech plastic packaging, such as multilayer films, which ensures a longer-term preservation of the food than a monolayer structure. From this point of view, multilayer films based on PLA and starch offer a useful alternative for food packaging purposes, taking advantage of the complementary barrier properties of both materials, and supplying the films with the high mechanical resistance of PLA. In this sense, amorphous PLA has demonstrated good adhesion properties with thermoplastic starch while the layer combination may be adequate for different food contact purposes, depending on the water or fat content of the food and the potential interactions of the packaging layers with these food components. Combining PLA and starch to obtain biodegradable food packaging materials represents good alternative means to reduce plastic waste. The blending of these polymers in adequate proportions with the incorporation of some compatibilizers has yielded high-performance films which can meet several packaging requirements. For instance, different PLA-starch ratios (70–30 or 80–20) with epoxidized soybean oil and maleic anhydride, or castor oil and hexamethylenediisocyanate offered good mechanical resistance and flexibility and could be potential replacers of non-degradable petrochemical polymers on the basis of their functional properties, polymer availability, adequate food contact properties and competitive cost. On the other hand, PLA-starch multilayer films exhibited a very good barrier capacity for water vapor and gases, while demonstrating reinforced mechanical resistance with respect to net starch films. Likewise, this strategy offers different possibilities for food contact depending on the layer combination (according to whether the food is more or less hydrophilic). Moreover, PLA or starch sheets can act as carriers of active compounds to produce antimicrobial or antioxidant packaging. According to the chemical affinity of the active, their incorporation into the hydrophobic (PLA) or hydrophilic (starch) layers can be designed to produce the most effective material.


According to Journal Citation Report by Clarivate Analytics , the impThis increase in IF, motivated us to make a brief analysis about the citations of the journal and its sources. We have observed that, 61% of all citations  appeared between January 2016 and May 2017. This analysis helped us to focus on and evaluate sources of citations.Between January 2016 and May 3, 2017, Balkan Medical Journal had been cited 338 times by 275 journals from 32 countries , Europe (33.5%) and Asia (12.7%) . BesidesIn other words, while journals from North America and Europe consist 75% of total journals, the percentage of journals from Balkans which cite Balkan Medical Journal is merely 2.2%. The citation-based regional analysis (338 citations) reveals us that the leading region which cites our journal is North America ; followed by Europe ; and Turkey  .We thought that it would be very interesting to check the origin of the first author of 338 papers. This analysis reveals us that the leading country is Turkey ; followed by Asia , Europe , North America  and Balkan countries . In the country-based analysis of the first author, Chinese authors are ranked as the third after their Turkish and American colleagues with 35 studies (10.4%) .In brief, 75% of the journals that cite the Journal are from North America and Europe; and similarly, 71.3% of 338 citations are from the journals published in these continents. However, the percentage of the North American and European first authors of 338 studies remain at 34.9% . On the One may argue that being indexed in PubMed Central in 2012 contributed and will contribute to increase the international visibility and accessibility of the journal as the last two years’ IF scores show. The rapid increase in submission allowed us to choose higher quality manuscripts. Despite recent improvements, quantity and quality of manuscripts originated from Balkan countries are still inadequate. We do believe in that, a close cooperation between team members leads the way to success. Therefore, we are in need to develop a scientific cooperation with Balkan countries. To realize this, Balkan Medical Journal aimed to constitute an efficient platform for sharing of scientific knowledge among Balkan countries.So, factors like being cited by authors from 51 countries with higher IF journals (even IF=21) are our fuel and source of motivation. Balkan Medical Journal has now completed 38 years of publication and is well recognized by international health community. We would like to thank and express our gratitude to all members of the Editorial Board, authors, reviewers, and publisher for their contribution and tremendous support.


Journal of Global Oncology (JGO), ASCO's new online-only, open-access journal dedicated to publishing the highest quality information concerning oncology research and care delivery in countries with limited health care resources.Welcome to the inaugural issue of 1 The presidential theme of past ASCO President Sandy Swain was “Building Bridges,” and global health was a major theme of ASCO's 2013 Annual Meeting. ASCO can indeed build bridges that serve to translate the work we do in the United States and other higher resource nations to improve cancer care in other countries. We can also build bridges that improve cancer care for health care disparity populations within developed nations by learning from the experiences of LMRCs that reflect the unique biologic and social attributes of these populations.2 This journal is a step toward building those bridges by enabling ASCO members to contribute to the vitally important and dynamic global conversation on health care, including access to care, quality of care, and perhaps most important, the design of systems of health care delivery that affect populations of patients with cancer.It is estimated that nearly two thirds of all cancer deaths occur in low- and middle-resource countries (LMRCs), and this is forecast to increase to 70% by 2030.Journal of Clinical Oncology and its sister Journal of Oncology Practice receive many submissions from around the world each year and have published special article series or collections about global oncology research in the past, there presently—or rather, surprisingly—is no journal that focuses solely on this important topic. JGO will fill this void. Our hope is that this journal will be the voice oncology professionals with a global connection or interest seek out for the latest practice-changing research and perspectives on cancer care, research, and care delivery issues unique to countries and settings with limited health care resources.International membership is the fastest growing category for ASCO. In fact, 31% of ASCO members practice outside the United States. Certainly all oncologists, regardless of where in the world they practice, share similar problems and concerns. US physicians have much to learn and gain from the research of our colleagues in other countries, and heretofore there had been a void in the literature of peer-reviewed research and expert commentary on the challenges of LMRCs. While ASCO's flagship JGO is eager to serve as a new home for this important work and serve as the channel by which to deliver it to all who are involved in cancer care. Because submissions from authors in LMRCs should be the lifeblood of JGO, ASCO will be providing more educational programs for researchers in these countries to support the conduct of meritorious clinical research and the successful submission of manuscripts to peer-reviewed journals.Already there is much good research coming from our international colleagues, but we lack a journal with a dedicated focus on global health. A manuscript with good information for the global oncology community may be rejected because of its limited relevance to the majority of readers of journals with a more general oncology focus. This means that vital information is not being published and getting to our oncology colleagues who need it. Annals of Oncology, where he served as Editor-in-Chief. Professor Kerr is a Fellow of the Royal College of Physicians, Honorary Fellow of the Royal College of General Practitioners, Fellow of the Academy of Medical Sciences, and Fellow of the European Academy of Cancer Sciences.Professor David Kerr of Oxford University will be at the helm of this new venture and brings a world of prestige and expertise to the position, having published many peer-reviewed articles and founded global oncology initiatives in India and Africa. He also has served on the editorial boards of several oncology journals, including the JGO's associate editors and editorial board members, who bring to the journal multidisciplinary expertise and firsthand clinical practice and research experience from around the world. We are honored and excited to watch this capable team grow ASCO's latest journal. We look forward to seeing Journal of Global Oncology discussed among colleagues at our various meetings around the world.As well, we are proud of JGO embarks on an exciting new venture. We look to you to send the journal your ideas for content. If JGO has not yet published something on a topic you consider current and vital to the internationally based oncologist, please let Dr Kerr know. This journal will look to international experts in the field of oncology to shed light on the challenges of cancer care and research in LMRCs through peer-reviewed original research and editorials. Some of these experts will be your friends. We ask you to encourage them to consider Journal of Global Oncology as a home for their important work.Thank you, reader, for sharing in this first issue. You are here as Journal of Global Oncology is indicative of ASCO's determination to take a lead role in oncology care across the globe by ensuring that oncologists in all settings are well-equipped with contemporary information and practice guidelines, and supports the Society's vision, as we all do, to help improve cancer care and delivery in LMRCs.As our population continues to diversify, so too will our oncology workforce and the patients they serve. The creation of


Drosophila midgut is a well-established model system for studying the mechanisms underlying ISC regenerative potential during aging. Here, we report the requirement of Drosophila phosphatidylethanolamine binding protein 1 (PEBP1) in ISC regenerative potential. We showed that PEBP1 was strongly expressed in enterocytes (ECs) of guts and its decrease with age and oxidative stress. Furthermore, the downregulation of PEBP1 in ECs accelerates ISC aging, as evidenced by ISC hyper-proliferation, γH2AX accumulation, and centrosome amplification, and intestinal hyperplasia. The decrease in PEBP1 expression was associated with increased extracellular signal-regulated kinase (ERK) activity in ECs. All these phenotypes by EC-specific depletion of PEBP1 were rescued by the concomitant inhibition of ERK signaling. Our findings evidence that the age-related downregulation of PEBP1 in ECs is a novel cause accelerating ISC aging and that PEBP1 is an EC-intrinsic suppressor of epidermal growth factor receptor (EGFR)/ERK signaling. Our study provides molecular insights into the tight regulation of EGFR/ERK signaling in niches for stem cell regenerative potential.The intestine is a high cellular turnover tissue largely dependent on the regenerative function of stem cell throughout life, and a signaling center for the health and viability of organisms. Therefore, better understanding of the mechanisms underlying the regulation of intestinal stem cell (ISC) regenerative potential is essential for the possible intervention of aging process and age-related diseases.  The intestine is not only essential for the uptake of nutrients but also acts as an effective barrier and protects organs from toxins and pathogens in the intestinal lumen. Furthermore, the intestine is a signaling center for the health and viability of the organisms . As the Drosophila is a well-established model system for studying ISC regenerative potential [Drosophila ISC are multipotent and give rise to both enterocytes (ECs) and enteroendocrine cells (EEs) either directly or through an intermediate state of enteroblasts (EBs) [The midgut of otential –9. Drosots (EBs) , 8. Thests (EBs) , 10, 11.ts (EBs) –15. ISC ts (EBs) –20. A rets (EBs) .Drosophila midgut [EC is an important niche in ISC homeostasis , 19, 22.a midgut , 18, 23.a midgut . HoweverThe EGFR/extracellular signal-regulated kinase signaling is a key pathway involved in intestinal homeostasis , 25, 26.Drosophila PEBP1 (CG18594), one of the six PEBPs, is highly expressed in larval and adult midgut [Phosphatidylethanolamine-binding protein 1 (PEBP1)/Raf kinase inhibitory protein (RKIP) plays an important role as a physiological inhibitor of EGFR/ERK signaling pathway in mammals . Loss ort midgut –39, but Drosophila PEBP1 plays a role as an EC-intrinsic suppressor of EGFR/ERK signaling and that the age-related decrease of PEBP1 in ECs accelerates ISC aging via EC death.Here, we show that Drosophila midgut by evaluating the transcriptional level of pebp1 gene with RT-qPCR. PEBP1 expression was found to be higher in the gut as compared with other tissues Myo1A which is active only in the mature EC [pebp1 mRNA greatly decreased by 0.27-fold in EC-specific PEBP1 knockdown guts as compared with the control guts  flies used as the age-related oxidative stress mimic model [− and Pdm1+ cells) was increased in 30- and 60-day-old guts  fly strains to distinguish mature polyploid ECs and mis-differentiated polyploid EC-like cells in aged guts. In comparison with the 5-day-old guts, the number of mature ECs with reduced expression of PEBP1 was increased in the 50-day-old guts   of catalase mutant flies  Figure . The rednt flies . These rDrosophila PEBP1 acts as a modulator of EGFR/ERK signaling, we investigated the correlation between PEBP1 expression and ERK activity in ECs using anti-phospho p44/p42 (pERK1/2) antibody as a marker for EGFR/ERK signaling [+ and Pdm1+ cells) was increased in 30-day-old and 60-day-old midguts  was increased in 50-day-old guts of esg-GFP fly strain, in comparison with 5-day-old guts . These results show that the downregulation of PEBP1 is related with an increase in ERK activity in ECs of aged guts.To evaluate whether ignaling . We foun midguts , arrows.s Figure . The incs Figure . Furtherts >+Myo1A and ts > PEBP1Myo1A flies. After 18h PQ treatment, the number of pERK+ ECs was increased in the guts of control flies (ts >+Myo1A), while the overexpression of PEBP1 in ECs reduced the number of pERK+ ECs  antibody as a mitosis marker. After 7 days of induction at 29° C, the number of PH3+ cells increased by up to 6.03-fold in EC-specific PEBP1 knockdown guts as compared with the control guts . The increase in the number of cleaved caspase-3+ ECs by PEBP1 knockdown was rescued by DIAP1 overexpression . The EC-specific overexpression of activated Raf in the guts resulted in an increase in the activity of ERK accompanied with the increased number of cleaved caspase-3+ ECs, enhanced proliferation, centrosome amplification, DNA damage accumulation in ISCs, and intestinal hyperplasia -H-x-H-R), which is a part of the ligand-binding pocket [Drosophila PEBP1 may be functionally similar to the mammalian PEBP1/RKIP, although it is still unclear how Drosophila PEBP1 negatively contribute to ERK signaling.The present study evidences that PEBP1 acts as an EC-intrinsic suppressor of ERK signaling. While the involvement of EGFR/ERK signaling in ISC homeostasis has been well defined , 20, 25,g pocket , althougg pocket . Thus, oWe also established that the downregulation of PEBP1 in ECs is a novel cause accelerating ISC aging. In the present study, the downregulation of PEBP1 and Raf overexpression in ECs induced age-related phenotypes of ISCs such as hyperproliferation, DNA damage accumulation and the increase in centrosome amplification, and hyperplasia. Our study also revealed that EC-specific depletion of PEBP1 induces EC death and our data in the experiment with concomitant DIAP1 overexpression revealed that the age-related phenotypes of ISCs by the downregulation of PEBP1 are associated with EC death. It is known that EC death is a common niche phenomenon promoting ISC proliferation through the secretion of several cytokines . IncreasHere, we evidence that the age-related downregulation of negative regulator is a cause of misregulated EGFR/ERK signaling during aging. In this study, we showed the activation of ERK in ECs of aged guts. We also showed the downregulation of PEBP1 in aged guts, indicating the age-related downregulation of negative regulator of EGFR/ERK signaling. In mammals, aberrant ERK signaling during aging has been known , 49. TheIn summary, our data strongly indicate that strong expression of PEBP1 in mature ECs of adult midgut is required for ISC homeostasis as EC-intrinsic inhibitor of EGFR/ERK signaling. Our study presents evidence showing that the age-related downregulation of PEBP1 in ECs is a novel cause accelerating ISC aging. In addition, our data provide molecular insight into the tight regulation of ERK signaling for intestinal homeostasis during aging.Drosophila stocks were maintained on standard cornmeal-sugar-yeast medium at 25° C under a 12 h light/dark cycle, as previously described [Oregon-R (OR) flies were used as wild-type. Catalase mutant (n1/+Cat) flies were obtained from a cross of OR males and n1/TM3Cat females. Delta-Gal4 and tubulin-Gal80 fly strains were combined to the different chromosome to generate teperature inducible ts;Delta-Gal4tubulin-Gal80 (tsDl) flies. For the induction of gene expression, conditional expression in adult flies using tstub-Gal80 was achieved by maintaining flies at 18° C throughout their developmental stage and shifting the 3-day-old flies to a higher temperature (29° C) for 3, 5, or 7 days. Esg-Gal4;UAS-GFP (esg-GFP) were obtained from B. Ohlstein [tsMyo1A-Gal4;UAS-GFP,tub-Gal80 (tsMyo1A) were obtained from B. A. Edgar [Delta-Gal4 were obtained from S. X. Hou [UAS-PEBP1 RNAi  and UAS-ERK RNAi (#35641) strains were obtained from the Vienna Drosophila RNAi Center . UAS-PEBP1 (#35835), gofUAS-Raf (#2033), UAS-DIAP1 (#6657), and tstub-Gal80 (#7108) strains were obtained from the Bloomington Drosophila Stock Center .All Ohlstein . Myo1A-G. X. Hou . UAS-PEBTM SYBR® Green Supermix . Data were acquired on a DNA Engine Chromo4 instrument (Bio-Rad). Cycling conditions were 30 s at 95° C, 30 s at 52° C, and 30 s at 72° C. Products were analyzed via melting curve analysis for 10 s at 95° C and 15 s at 65° C, followed by a temperature ramp from 65 to 95° C (0.1° C/s) and continuous fluorescence recording. Oligonucleotide sequences for the analysis of pebp1 [rp49 [Total RNA was isolated from adult tissues using TRIzol reagent  according to the manufacturer's instructions, and 1 μg total RNA was reverse transcribed using M-MLV reverse transcriptase . Quantitative real-time PCR was performed using iQof pebp1  and rp49p1 [rp49  were preImmunostaining was performed as previously described , 19. IntMidguts were dissected, fixed in 4% paraformaldehyde at room temperature for 30 min, and infiltrated overnight with 20% sucrose at 4° C. After flash-freezing in Tissue-Tek optimal cutting temperature (OCT) medium , 10 μm sections were cut on a cryostat  at −20° C. Sections were blocked in 5% BSA (Sigma-Aldrich) for 1 h and incubated overnight with primary antibodies. The samples were washed in PBST and incubated with secondary antibodies conjugated to a fluorescent dye at 25° C for 1 h, followed by their mounting with Vectashield (Vector Labs) and analysis using a Zeiss AxioSkop 2 Plus microscope (Carl Zeiss Inc.).For the immunohistochemistry, the following primary antibodies were used at indicated dilutions: mouse anti-Delta , 1:100, mouse anti-Prospero , 1:100 and mouse anti-Armadillo (Arm), 1:100 ; mouse anti-GFP and rabbit anti-GFP, 1:500 ; rat anti-GFP, 1:500 ; rabbit anti-γH2AvD, 1:1000 ; mouse anti-γ-tubulin, 1:500 (Sigma-Aldrich); rabbit anti-phospho-histone H3, 1:500 ; rabbit anti-cleaved caspase-3, 1:100 and rabbit anti-p44/p42 mitogen-activated protein kinase , 1:100 ; mouse anti-Pdm1, 1:100 ; rabbit anti-Pdm1, 1:100 (from Y. Cai); and rabbit anti-PEBP1, 1:200 (developed in this study). The following secondary antibodies were used at indicated dilutions: fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit, 1:500, Cy3-conjugated goat anti-rabbit, 1:500, FITC-conjugated goat anti-mouse, 1:500 and Cy3-conjugated goat anti-mouse, 1:500 . Nuclei were labeled with 4,6-diamidino-2-phenylindole (DAPI), 1:500 (Molecular Probes). Visceral muscle (VM) was labeled with rhodamine phalloidin (RP), 1:500 .Drosophila PEBP1 (CG18594) antibody was produced by NovoPro Bioscience Inc. . PEBP1 peptide was synthetized by solid phase using Fmoc chemistry. The purity and amino acid composition were confirmed by high-performance liquid chromatography (HPLC) and electrospray ionization mass spectrometry (ESI-MS). The peptide (DQPTVVFDAEPNSLY-Cys) was coupled to keyhole limpet hemocyanin (KLH) through thiol. KLH-coupled antigens (about 1 mg) were mixed with an equal volume of complete Freund adjuvant and injected subcutaneously into two New Zealand rabbits. Boosts (0.5 mg) were given 2 weeks later at an interval of 1 week. Immunopositive sera were monitored by enzyme-linked immunosorbent assay (ELISA) with KLH-coupled antigens as screening peptides. After seven boosts (about 9 weeks), the anti-serum was purified with a peptide-affinity column.ts>+Myo1A and ts>PEBP1 RNAiMyo1A flies after induction of 5 days at 29° C. Total protein from these guts were extracted using a lysis buffer  with a protease inhibitor . Protein quantification was done with Bradford reagent (Bio-Rad) before equal amount of protein was loaded. The whole gut extracts were separated by sodium dodecyl sulfate (SDS)-polyacrylamide gels containing 15% acrylamide and the protein bands were transferred onto polyvinylidene fluoride membranes . The blotted membranes were blocked with 1×TBST containing 5% skim milk, followed by their overnight incubation with the rabbit PEBP1 antibody (1:2000) and goat anti-actin antibody  at 4° C. After washing, the membranes were incubated with horseradish peroxidase (HRP)-conjugated secondary antibodies (1:5000) for 2 h at room temperature. The protein bands were detected with enhanced chemiluminescence (ECL) western blotting detection reagent .A total of 30 guts were dissected from 5-, 30- and 60-day-old flies, or ts>+Myo1A and ts>PEBP1Myo1A flies after induction of 5 days at 29° C were starved for 2 h and exposed to 10 mM paraquat (Sigma-Aldrich) in 1% sucrose media for 18 h. During paraquat treatment, ts>+Myo1A and ts>PEBP1Myo1A flies were maintained in a 29° C incubator for the activation of the gene expression of UAS-Gal4 system. Following paraquat treatment, guts were subjected to immunostaining.Paraquat  feeding assay was conducted as previously described . Five-daFlies were irradiated with a single dose of 20 Gy using a gamma irradiation machine  at a dose rate of 2.25 Gy/min. A continuous dose was administered to the whole body of flies in the irradiation chamber at room temperature. After irradiation, irradiated flies were maintained in a 29° C incubator for the activation of the gene expression of UAS-Gal4 system.+ cells and supernumerary centrosomes (≥3) in mitotic ISCs were determined in whole midguts, defined from R1 to R5 by Buchon et al [+ ECs were counted from the posterior midgut R5 region under microscopic fields at a magnification of 400×. Data are expressed as mean ± standard error of the mean (SEM) and the statistical significance of differences was determined using the Student's t-test . A value of p < 0.05 was considered statistically significant. GraphPad Prism 6  was used for the analysis of variance.The numbers of PH3on et al . The num


P ≤ 0.001) for all traits analyzed. Kiroba recorded high starch yields of 17.4, 12.7, and 8.2 t ha−1 at Chambezi, Amani, and Magadu, respectively. Kilusungu recorded highest cyanide content of 300–400 ppm across all locations but Kiroba recorded highest values of 800 ppm, 15 months after planting at Chambezi. Genotype by environment (GGE) biplot analysis revealed that Kiroba was a superior cultivar in terms of starch yield. Kilusungu recorded highest cyanide content and average starch yield, therefore suitable for use in starch production. The study confirmed effect of genotype and genotype by environment interaction, Kiroba cultivar was superior in terms of starch yield and maximum starch yield was obtained at 9 months after planting. Nyamkagile and Kibandameno had the lowest cyanide content across all environments.High starch yield is the most important trait for commercialized cassava starch production. Furthermore, cyanide present in cassava roots poses a health challenge in the use of cassava for food. Cassava genotypes have varying maturity periods that are also environmental dependent. This study aimed at identifying suitable cultivars and optimum time of harvest to maximize starch production across three environments. The study found significant difference between genotypes, locations, harvest period, and all the interactions ( Starchy tuberous roots of cassava provide food for people in sub‐Saharan Africa and Latin America  Santisopasri et al.  working ,Cyanide is produced by plants as a by‐product of ethylene metabolism, or as reduced form of nitrogen storage and defense against attack by herbivorous  cultivars as protection from predators and thieves . Three healthy plants were uprooted from each plot, marketable roots were selected, counted and weighed (SRM). For starch and dry matter analysis, roots were brought immediately to the laboratory of Food Science and Technology, Sokoine University of Agriculture (SUA).Trials were set at Chambezi (Bagamoyo), Amani (Muheza), and Magadu  from April 2012 up to July 2013 Table . Soil saFSRY (t/ha) was calculated from the formula:SRM = Storage Root Mass.1) were put in preweighed petri dishes (W0). The samples were oven dried at 60°C for 72 h, and weighed (W2) after removal from the oven. DM was calculated as follows;Cassava root dry matter (DM) was determined according to Benesi et al. . Fresh c. Exactly 500 g of fresh peeled cassava from three root samples were chopped using water as extraction solvent in a laboratory blender. The tissues were broken and fiber separated by sieving through double layer of cheese cloth. After washing several times, starch was oven dried at 50–55°C for 72 h and then weighed (Wo).Starch extraction was conducted following the method by Benesi et al.  with sliSC (g/kg) was calculated as = Wo/500*1000−1.Starch yield was obtained as the product of fresh cassava yield and starch content expressed as t haStarch yield = SC*FSRY.Cyanide content was determined following the picrate paper method as illustrated by Bradbury . Root saPpm = mg HCN equivalents/kg fresh root.Data analysis was done using GENSTAT 11 program and Analysis of Variance (ANOVA) for all the traits was performed. When the effects of genotype by environment interactions were significant, further analysis was done using genotype and genotype by environment (GGE) biplot procedure on starch yield and cyanide content. Genotype by environment biplots analysis was performed to determine landrace's superiority, stability, and also which‐won‐where in relation to starch yield and cyanide content . At Chambezi, there was a yield increase from 9 to 15 MAP for three landraces , with Msenene showing the highest increase. The highest yield (39 t/ha) was recorded at 15 MAP from Msenene. At Amani, there was a yield increase from 9 to 15 MAP for four landraces . The highest yield (25 t/ha) was recorded at 15 MAP from Kibandameno. At Magadu, there was also a yield increase from 9 to 15 MAP from four landraces . The highest yield (12.6) was recorded at 15 MAP from Kiroba.The mean fresh root yield at each site and harvest round are given on Table P ≤ 0.001) was observed. Increase in DM was evident from 9 MAP to 12 MAP but decrease at 15 MAP, a trend observed at all sites. However, Kilusungu recorded highest DM at 15 MAP at Bagamoyo site. Highest DM was recorded at 15 MAP from Kibandameno at Chambezi and Amani site recorded highest overall average dry matter and starch content (Table The means for root dry matter content at each site and harvest round are given on Table nt Table  in most P ≤ 0.001). Nyamkagile had cyanide content below 30 ppm at Magadu. Furthermore, Kibandameno also had low cyanide content at Bagamoyo site at 9 and 12 MAP. The trend showed that each landrace gave different response at each harvesting time and it varied with location. Nyamkagile was the only landrace which showed decrease in cyanide content with age. However, the trend and cyanide concentration differed for each environment as demonstrated by significant GEI (P ≤ 0.001) (Table Variations in cyanide content Table  were hig1) Table .The analyses of variances (ANOVA) for fresh root yield, starch content, starch yield, and cyanide content across sites and harvesting rounds Table  show sigIn terms of starch content, the trend per site was Amani > Magadu > Chambezi. For fresh root yield, Chambezi > Amani > Magadu  of the mean square difference between the performance of a variety and the best variety within a given environment, measuring mean performance and stability simultaneously. GGE biplots enables visual comparison of the location, genotype, and their interaction.The GGE biplot analysis showed that PC1 and PC2 accounted for 91.4 and 8.12%  variation, respectively, for starch yield . A convex hull with three vectors was formed with Kiroba, Nyamkagile, and Kalolo as the vertex cultivars Fig. B. KirobaThe GGE biplot analysis for cyanide content showed that PC1 and PC2 accounted for 77.1% and 13.5%, respectively . The dry period that existed between January and March (9–12 MAP) caused low carbohydrate partitioning in the storage root  between landraces and GEI, therefore confirming previous studies. McMahon et al.  on (A) fresh root yield (B) dry matter content (C) starch content (D) cyanide content, of six cassava landraces across three sites.Figure S2. GGE Scatter plot showing discrimination and representativeness of the environments (sites) using the starch yield data of the six cassava landraces.Figure S3. GGE Scatter plot showing discrimination and representativeness of the environments (sites) using the cyanide content of the six cassava landraces.Click here for additional data file.


Inspection of different density gradients showed a continuous increase in ‘heavy’ 16S rRNA in caecal samples over the course of the experiment. Sequencing analyses of unlabeled and labeled 16S amplicons particularly suggested a group of unclassified Clostridiales, Dorea, and a few other taxa  to be most actively involved in starch assimilation in vivo. In addition, metabolic product analyses revealed that the predominant 13C-labeled short chain fatty acid (SCFA) in caecal contents produced from the [U13C] starch was butyrate. For the first time, this study provides insights into the metabolic transformation of RS by intestinal bacterial communities directly within a gut ecosystem, which will finally help to better understand its prebiotic potential and possible applications in human health.Resistant starch (RS) is the digestion resistant fraction of complex polysaccharide starch. By reaching the large bowel, RS can function as a prebiotic carbohydrate, i.e., it can shape the structure and activity of bowel bacterial communities towards a profile that confers health benefits. However, knowledge about the fate of RS in complex intestinal communities and the microbial members involved in its degradation is limited. In this study, 16S ribosomal RNA (rRNA)-based stable isotope probing (RNA-SIP) was used to identify mouse bowel bacteria involved in the assimilation of RS or its derivatives directly in their natural gut habitat. Stable-isotope [U The large bowel of humans and other mammals harbors a complex and highly diverse microbiota . AlthougBifidobacterium, Ruminococcus, Lactobacillus and Roseburia due to ingestion of RS -labeled potato starch, a source of RS, in vivo in a complex caecal community resident in mice. We observed incorporation of the 13C-label into bacterial 16S rRNA only 2 h after oral administration of [U13C] starch to the mice. Phylogenetic analysis of unlabeled and labeled 16S rRNA pools revealed slight differences in the caecal microbiota structure, indicating microbial taxa involved in starch degradation. In addition, metabolite screening by gas chromatography-mass spectrometry (GC-MS) analysis showed label incorporation mainly into butyrate. To the best of our knowledge, our results represent the second report worldwide using RNA-SIP under the in vivo conditions of a vertebrate intestinal system  starch extracted from raw potatoes  via oral gavage into the stomach. The placebo control group received 0.5 mL of a PBS solution (pH 7.0) without starch. Animals were sacrificed at 0 h (placebo control group) or 2 h and 4 h after starch administration by cervical dislocation and the entire gastrointestinal tract (GIT) was immediately dissected post mortem using sterile surgical instruments. The caecum section was opened individually, and contents were collected from each animal. Samples were resuspended in 1 mL of sterile RNAlater  and stored at −80 °C until further analyses.Mice were randomly assigned into three groups of three animals each. At the start, animals of two groups received 0.5 mL of a phosphate-buffered saline solution  suspended with ~0.4 g of uniformly (98%) isotope-labeled  starch assimilation, 16S rRNA gene amplicon libraries were constructed from density-separated and reverse-transcribed RNA obtained from selected SIP fractions. SIP fractions 8–10 contained the bulk of 16S rRNA present in samples from control mice that did not receive any [U13C] starch (0 h). These fractions were considered to represent unlabeled bacterial populations and were thus defined ‘light’ fractions. In caecal samples of mice fed with [U13C] starch, gradient fractions 5–7 contained the bulk of RNA, which was consequently considered to be ‘heavy’, i.e., 13C-labeled, representing bacteria that particularly assimilated the [U13C] starch and sequestered the isotope label in their RNA.For the identification of distinct bacterial taxa involved in  starch shifted towards the higher density fractions 5–7 (~1.817–1.802 g mL−1) with the maximum amount of RNA detected in fraction 7 after 4 h. Furthermore, although after 2 h the majority of RNA still accumulated in fraction 8 and 9 (~1.794 and ~1.786 g mL−1), fraction 7 (~1.802 g mL−1) contained more RNA than control fraction 7 .Lachnospiraceae, unclassified Clostridiales and Ruminococcaceae 16S rRNA gene sequences relatively dominating the ‘light’ as well as the ‘heavy’ communities  (p = 0.038) as well as between the different sampling times (p = 0.023) were detected  . In addidetected .13C] starch, sequencing profiles retrieved form ‘heavy’ and ‘light’ SIP fractions at each sampling time were screened for relative differences of individual taxa; 2 h after administration of [U13C] starch, taxa that showed significantly increased proportions in ‘heavy’ fractions compared to the corresponding ‘light’ counterparts included unclassified Clostridiales (p = 0.03) and Dorea spp. (p = 0.03). Taxa that tended towards significant increases included Bacteroidaceae (p = 0.07), Bacteroides spp. (p = 0.06) and Turicibacter spp. (p = 0.08). All these groups also showed higher abundances in the ‘heavy’ RNA 4 h after starch administration, however, not with statistical significance (To identify bacteria possibly involved in the degradation of the  starch dose , showed inter-animal variations, in each mouse, butyrate was the predominant labeled fermentation product, followed by lactate and acetate 2 h after the administration of starch. During the course of fermentation, isotope-labeled butyrate as well as acetate were consistently produced from the [U13C] starch, while amounts of 13C-labeled lactate concentrations declined after 2 h.To gain further insight into the metabolic activity of the caecal microbiota, GC-FID and GC-MS analysis was performed in supernatants of caecal contents to monitor Bifidobacterium, Akkermansia, Allobaculum, Lactobacillus, Ruminococcus, Prevotella and several other groups -labeled potato starch, a source of RS, as a substrate for the caecal community using mice as model system, and the use of RNA-SIP in combination with high-throughput 16S rRNA sequencing and GC-MS analysis to gain phylogenetic information about the bacteria involved in [U13C] starch degradation in situ. In a previous in vitro study, we had used RNA-SIP to identify bacterial groups from faecal samples of mice that were involved in the in vitro assimilation of [U13C] starch  starch . To the ] starch , and the13C]RNA from unlabeled background RNA [13C] starch-fed mice towards higher densities, 2 h and 4 h after provision of the labeled starch clearly indicated [U13C] starch utilization and incorporation of 13C into bacterial 16S rRNA. After 2 h of feeding, labeling of RNA was less pronounced, which might be explained with the digestion-resistant nature of RS, the presence of competing  starch. Interestingly, only a few of them are related to well-known starch degraders.Screening of 16S amplicon sequencing profiles revealed only a limited number of taxa that were significantly increased in abundance in ‘heavy’ vs. ‘light’ gradient fractions, thereby suggesting involvement in the assimilation of the [UBacteroidaceae and Bacteroides spp. were relatively enriched in ‘heavy’ RNA, isolated 2 h after feeding starch. These taxa comprise a particular high number of members that are associated with the utilization of various types of starch and are, therefore, considered to be one of the major amylolytic species in the human colon [Bacteroides spp. is mediated through a well-characterized starch binding and degradation system referred to as starch utilization system (SUS) [Bacteroides spp. might have functioned as initial starch degraders here, producing smaller hydrolysis products, e.g., maltose and glucose that served as substrates for other taxa such as Dorea spp. and unclassified Clostridiales, which became significantly labeled during the experiment. This is in line with a previous report demonstrating that gut Bacteroidales significantly contributed to cross-feeding in intestinal ecosystems [With limited significance, an colon ,51,52. Sem (SUS) . Hence, osystems . Notablyosystems . Howeverosystems ,57.Dorea is one of the most dominant human gut genera [Dorea portions upon a diet enriched with RS were recently reported in mice [Dorea species, including Dorea formicigenerans, to which the most abundant OTU from this group was affiliated, are not known to be capable of starch hydrolysis so far [Dorea ssp. here represent so-far uncultured species, being capable of starch hydrolysis, or the observed labeling results from cross-feeding on metabolites released from the labeled starch by other bacteria or by the host.t genera . Increas in mice , suggest in mice ,60,61. Ns so far . Either,Turicibacter spp. or Clostridium lactatifermentans [Turicibacter spp. were also shown in the hindgut of pigs fed raw potato starch [Clostridiales’ appear to be affiliated with Lachnospiraceae, whose members are known for their capacity to degrade several plant-derived polysaccharides including starch [Both explanations also hold true for the other taxa that were found to be enriched in the ‘heavy’ RNA fraction, as many of them were related to taxa so far not known as starch degraders, such as rmentans ,64. Inteo starch  and two g starch .Clostridium cluster IV (Ruminococcaceae family) and Clostridium cluster XIVa (Lachnospiraceae family) [Clostridiales and the Lachnospiraceae family, which also became significantly labeled during the experiment.RS is well-known to significantly increase SCFA production in vitro and in vivo ,68,69. I family) , and/or  family) ,74,75. S13C-labeld, which was now confirmed for Bacteroides and unclassified Clostridiales also under in vivo conditions, thereby strongly suggesting an actual involvement of these groups in intestinal starch degradation.The data obtained here show similarities in comparison to our recently published in vitro data on the fermentation of potato starch by a mice fecal microbiota . While iBacteroides and Clostridiales were confirmed as initial starch degraders in the caecal microbiota of mice, while other bacteria related to Clostridiales, Dorea, and Turicibacter might have played a role as cross-feeders on metabolic products, although a direct involvement in starch degradation cannot be ruled out. Clearly, further studies are needed to verify the role of the bacteria suggested as starch degraders and cross-feeders here in more detail. These studies should account for the limitations of our pilot study, i.e., include, for instance, more test and control animals, more sampling points (to address the effect of cross-feeding in more detail), longer incubation times (to achieve a stronger labeling of RNA and metabolites) and 12C-controls . Finally, SIP might be used in a more quantitative way, as recently introduced by Hungate and colleagues [Collectively, our data show that the RNA-SIP approach is generally suited to link the structure and function of intestinal microbiotas also under the in situ conditions of mice intestinal tracts. Heavier, i.e., presumably isotopically labeled RNA, could be extracted only 2–4 h after having fed labeled potato starch to the animals. In addition, a few, but statistically significant differences in microbial community composition between labeled and unlabeled RNA fractions were shown, probably indicating starch-degrading bacteria from the intestinal tract. Bacteria affiliated with lleagues , albeit


Biochemia Medica 2017;27 . In our analysis of Croatian open-access journals in the Hrčak repository , we presented the data on number of journals from Hrčak subsidized by the MSE in 2016 .In our article „Does small equal predatory? Analysis of publication charges and transparency of editorial policies in Croatian open access journals“ published in 2017 June issue of the Because of the inconsistencies in the identification of the journals, we miscategorized 2 out of 175 journals receiving the financial support from MSE as not being included in the Hrčak repository. The correct statement should be that all journals supported by the MSE were in the Hrčak repository. This correction in no other way influence the results and the message of our study.https://mzo.hr/sites/default/files/dokumenti/2017/02/znanstveni_casopisi_i_casopisi_za_popularizaciju_znanosti_odobreni_u-2016_godini.pdf) and the list of Hrčak journals, available at http://hrcak.srce.hr/stat.php. The MSE list includes the name of the publisher, journal title, scientific field, requested and approved financial support in 2016. The list does not include the International Standard Serial Number (ISSN), which is the unique identifier of a journal. The list at Hrčak includes the journal title, status in Hrčak, ISSN (online), ISSN (print), URL of the journal, coverage, number of articles for different criteria, date of journal inclusion, date of the publication of the last issue and date of the last Hrčak upload. This list does not include subtitles of the journal, which are a part of the bibliographic records and are visible at other Hrčak web-pages. As both lists have journal titles, we presumed that automated comparison of the journal titles would give the correct matching of the titles in the two lists. During our analysis, two journal titles were not matched because journal titles differed and we stated in the published article that two journals that received the financial support from the MSE were not included in the Hrčak repository. Since the financial support of the journals was not the focus of the study, we did not perform a check for each of the individual journals.For our analysis, we have used the list available at the MSE site (Radovi Zavoda za hrvatsku povijest (without ISSN and subtitle), carries the main title of Radovi in the Hrčak repository . The same journal title Radovi is also present in the catalogue of the National and University Library (NUL) and their ISSN office, and on the journal cover page itself. The subtitle in Hrčak is Radovi Zavoda za hrvatsku povijest Filozofskoga fakulteta Sveučilišta u Zagrebu. Scopus and Web of Science Core Collection (WoSCC) databases index the journal as Radovi Zavoda za Hrvatsku Povijest with the correct ISSNs. On its own website, the journal cites the title Radovi Zavoda za hrvatsku povijest but the issues themselves are listed under the title RADOVI.The first journal, present on the MSE list of funded journals under the title Hrvatski časopis za odgoj i obrazovanje (without ISSN and without subtitle), carries the main title of the Croatian Journal of Education in the Hrčak repository  and the subtitle Hrvatski časopis za odgoj i obrazovanje. In the NUL catalogue, the journal has the Croatian title Hrvatski časopis za odgoj i obrazovanje. WoSCC indexes it as the CROATIAN JOURNAL OF EDUCATION-HRVATSKI CASOPIS ZA ODGOJ I OBRAZOVANJE and Scopus indexes it as the Croatian Journal of Education, both with the correct ISSNs.The second journal, present on the MSE list of funded journals as Inconsistent presentation of journal information in the online environment is not rare in the global research community and has been discussed by several studies (


The Brazilian Journal of Cardiovascular Surgery (BJCVS) is grateful for the reviewers,listed below, which collaborated in this edition. Without their work would be impossibleto keep the high scientific standard of our journal.Carla TanamatiCharles Simão FilhoEnio BuffoloFrancisco Fernandes Moreira NetoGilberto Venossi BarbosaGiovani Jose Dal Poggetto MolinariJosé Glauco Lobo FilhoLeonardo Andrade MulinariLindemberg Da Mota Silveira FilhoLuciano AlbuquerqueLuiz César Guarita SouzaRicardo de Carvalho LimaRodrigo MilaniRui M. S. AlmeidaTomas SalernoUlisses A. CrotiVictor Rodrigues Ribeiro Ferreira


High capacity and low capacity running rats, HCR and LCR respectively, have been bred to represent two extremes of running endurance and have recently demonstrated disparities in fuel usage during transient aerobic exercise. HCR rats can maintain fatty acid (FA) utilization throughout the course of transient aerobic exercise whereas LCR rats rely predominantly on glucose utilization. We hypothesized that the difference between HCR and LCR fuel utilization could be explained by a difference in mitochondrial density. To test this hypothesis and to investigate mechanisms of fuel selection, we used a constraint-based kinetic analysis of whole-body metabolism to analyze transient exercise data from these rats. Our model analysis used a thermodynamically constrained kinetic framework that accounts for glycolysis, the TCA cycle, and mitochondrial FA transport and oxidation. The model can effectively match the observed relative rates of oxidation of glucose versus FA, as a function of ATP demand. In searching for the minimal differences required to explain metabolic function in HCR versus LCR rats, it was determined that the whole-body metabolic phenotype of LCR, compared to the HCR, could be explained by a ~50% reduction in total mitochondrial activity with an additional 5-fold reduction in mitochondrial FA transport activity. Finally, we postulate that over sustained periods of exercise that LCR can partly overcome the initial deficit in FA catabolic activity by upregulating FA transport and/or oxidation processes. Our bodies consume carbohydrates, fats, and amino acids as fuels, utilizing various catabolic pathways to transfer the energy required for normal physiological functions. The way these pathways function can have an important impact on overall health. While most catabolic pathways are known, we are still striving to understand how these pathways interact, are controlled, and change during exercise and in disease. Here, we have used computer modeling as a tool to understand fuel utilization differences during exercise for two animal models. High capacity running rats (HCR) were used as a healthy, fit cohort, and low capacity running rats (LCR) were used as a sedentary and disease-prone cohort. Our computer model results show that the HCRs are superior at fat utilization compared to LCRs because of their increased ability to transport and catabolize fatty acids. We postulate that these differences depend on exercise intensity and duration, such that longer acclimation periods may minimize fuel utilization differences between these rats. Fuel selection, the balancing of glucose, fatty acids, and amino acid utilization to match the ATP demand, is a hallmark of healthy metabolism . It has A popular mechanistic view of fuel selection centers on the Randle cycle , 21, whiHigh and low capacity running rats ,  , Methods network .Initial FA, was simplified to be only palmitate (C16 acyl chain length), and glucose was the only carbohydrate considered. The computed 87 internal reaction fluxes, with time, are shown for HCR  and LCR To test the stability of the HCR and LCR solutions , we deteX). Furthermore, ATP demand was simulated based on the interpolated rate from the constraint-based solution , plasma lactate  are shown in The predicted ratios of HCR and LCR enzyme activities , such as that shown in 3 parameter sets capable of fitting each data set. We then took the ratio of all possible combinations of parameter sets between HCR and LCR yielding 106 ratios (To obtain a distribution of activity ratios (X6 ratios  for eachAlthough a number of enzyme activities (model adjustable parameters) tend to fall within a relatively small range  to fit t3 parameter sets with respect to the simulation of the respiratory quotient (JCO2/JO2). The respiratory quotient is typically considered to be a good indicator of fuel selection based on stoichiometric utilization of O2 and production of CO2 for different substrates. The results of this calculation yielding 103 sensitivity coefficients for HCR , hexokinase (HK), and the combination of complex 3 and 4 (Complex 3+4) activity Figs  and 10. To help combine the information collected by computing numerous enzyme activity ratios  and sensThe enzyme score calculation reveals higher importance for glycolytic enzymes for LCR and FA catabolic enzymes for HCR, although our minimal approach to fitting LCR data  showed tTo theoretically estimate the FA utilization capacity of HCR and LCR, we decreased glycolytic activities by several orders of magnitude until no appreciable glycolytic flux was observed. Without the ability to synthesize ATP via glycolysis, and eliminating ATP buffering by creatine kinase, FA utilization is the only means to sustain ATP synthesis in our model. After decreasing these activities, an increasing linear ATPase rate was imposed until the ATP synthesis capacity was overwhelmed by ATP demand, at which point ATP concentration collapses . With glTo compare HCR and LCR fuel selection with normalized exercise intensity, we plotted the respiratory quotient (RQ) data for HCR and LCR, as a function of their %VO2max . These dc). A transition constant (Tc) of 0.15 and 0.1 were used for HCR and LCR, respectively. A simulation time of 2 minutes was used to simulate the transient data . We also asked the question if a model completely based on mass-action kinetics and thermodynamic constraints, lacking detailed enzyme kinetic modeling, could still reproduce fuel switching dependent on ATP demand.Constraint-based modeling, rooted in flux balance analysis , was firIt has been previously shown that LCR have similar mitochondrial density in red gastrocnemius muscle , 37, butOur model does not distinguish between enzyme activity effects induced by PTM or expression levels, however, it does support the need to invoke combined changes in both total mitochondrial activity  and a specific decrease in FA transport. Additional decreases in FA β-oxidation enzyme activities  were alsOur model is simplified in that it does not account for amino acid catabolism, although amino acid data were provided in the data set we analyzed . This siDespite the need to decrease HCR FA enzyme activities to simulate LCR data, it is important to understand that these data  were colThus, it appears that the difference between LCR and HCR FAO diminishes with sustained normalized exercise intensity . This rein vivo, but does show that simple kinetic modeling can effectively capture this phenomenon without involving these regulatory processes.Finally, it is interesting that the simple kinetic model developed here, for the HCR rat, can switch between FA and carbohydrate substrates in a physiological manner, as demonstrated by simulating the metabolic response to exercise. The model does not include allostery, such as required for the Randle cycle , 21 or cS  is the stoichiometric matrix and  fGoldberg  databaseGoldberg  server. ussed in ), where The resulting concentration vector was slightly adjusted to match the data shown in C) is the concentration vector, (P) is a partition matrix, (S) is the stoichiometric matrix, and  is the calculated reaction flux vector. The partition matrix is a diagonal matrix that compartmentalizes the system into extracellular, cytosolic, and mitochondrial spaces with partition coefficients of 0.2, 0.75, and 0.05, respectively, based on the fractional volume of skeletal muscle /[HCR (metabolites)], therefore metabolite concentrations that are greater in LCR become much more visible.(TIF)Click here for additional data file.S1 Matlab CodeThis code also contains the code required to simulate the model as is needed to generate several figures.(ZIP)Click here for additional data file.S1 Matlab Figures(ZIP)Click here for additional data file.


By 2030, noncommunicable diseases (NCDs) will be the leading cause of death in every region in the world. While law and policy have an important role to play in curbing this pandemic, our current understanding of how they can most effectively be used is still limited. This contribution identifies a number of gaps in current research and insists on an interdisciplinary research agenda between law, health science and international relations aimed at designing concrete proposals for laws and policies to curb the NCD pandemic, both globally and domestically. In 2012, 38 million people died from noncommunicable diseases (NCDs), accounting for 68% of total deaths globally [The scale and universality of the problem make NCDs a pandemic phenomenon that requires a powerful international response. In this response, prevention is key, because much of the global NCD burden (40%) is linked to four “modifiable behavioral risk factors” that affect many countries: tobacco use, unhealthy diets, physical inactivity and harmful use of alcohol  adopted a set of nine voluntary targets on NCDs to be attained by 2025 . This ‘2Researchers from various disciplinary angles have engaged with NCDs and have made a meaningful contribution to the development of effective NCD laws and policies –8. HowevWhile law and policy interventions have proven successful in curbing NCD incidence already, we need to learn more about the success factors that can be attributed to recent domestic and international instruments. To achieve this goal, we argue in favour of a truly integrated research agenda spanning health science, domestic and international legal scholarship, and international relations theory. In practice, this means that  law and policy-oriented NCD-research needs to build more on health science and international relations research, but also that health science can generate insights on the success of policy interventions. This is further explained below. In addition, domestic and international policy makers should better understand the important role of legal arrangements, while law-makers can learn from domestic and international policy realities and other challenges to which legal interventions need to respond.The overall aim of our proposed integrated research agenda is to develop a comprehensive toolbox that can guide law and policy making, and assists identifying ‘good practice’ laws and policies for domestic governments to adopt . This tyDomestically, there is increasing evidence that taxes on sugar sweetened beverages result in reduced consumption of these beverages, and that subsidies for fresh fruits and vegetables can increase fruit and vegetable consumption . MoreoveAt the same time, knowledge about the various regulatory options available to lawmakers and policymakers can be greatly improved . EvidencTo identify effective law and policy options, in particular ‘best practices’, coordination between health science and law is crucial. Evidence of current patterns of NCDs, and interventions related to them, creates insights into which laws and policies can best be put into place. It also reveals how the implementation of laws and policies is succeeding. In respect of evidence, the estimate that in Europe obesity accounts for about 65–80% of new cases of Type 2 Diabetes is an important signal to legislators to reflect on the need to regulate risk factors such as unhealthy diets, which may be achieved by more effective regulation of certain foods . At the should and can attempt to influence the behavioral and consumption patterns of individuals directly through regulation [Despite overwhelming evidence of the scale of the NCD pandemic, and the importance and possibilities of curbing NCDs through prevention, we are aware that attempting to change lifestyle raises the difficult question as to whether and how national and international authorities gulation . First, gulation .Second, there is evidence that nudging, through regulation alone, may be insufficient because there are important underlying social determinants of health. For example, poverty, poor family relations, unemployment, and a lack of adequate education may increase engagement in certain risk factors , 6. SuccLastly, the practical and social role of companies should not be under-estimated. They heavily control essential aspects such as marketing, a product’s ingredients and presentation, and the availability of alternative products. These are important factors that influence people’s free choice which cannot be ignored. Moreover, while marketing bans or sales points restrictions may offer powerful tools to counter some of these forces, typically, tobacco, alcohol and food and beverages’ companies also have large budgets for lobbying and influencing markets and policy-makers in profound, if not subversive, ways. This type of influence may also have to be reigned in, nationally and internationally.Despite the fact that NCDs pose the largest threat to future global health, they remain an under-emphasized area of inter-disciplinary research . We arguAnother under-researched dimension of NCDs concerns the question of how social and legal change can be brought about at both international and domestic levels. International relations’ research can give a crucial insight into the political processes required for new effective NCD laws and policies to be adopted. A close interaction between law and political science is required to study these mechanisms.International laws and guidelines are essential for global agenda-setting in relation to NCDs and to ensure a comprehensive international response to the rising NCD pandemic. They present an important driving force for national authorities to prioritize certain health concerns and arrive at better regulation. So far the only legally binding international instrument that addresses a behavioral risk factor is the WHO’s Framework Convention on Tobacco Control (FCTC). This influential treaty, which was adopted in 2003 and is currently ratified by 179 countries, has led to a tightening of many domestic tobacco laws and to several domestic court cases addressing the harmful effects of tobacco . There hAn important related question is whether new international standards should be pursued through treaty-making, or whether “soft-law” instruments in the form of Guidelines, Standards, Codes of Conduct or Action Plans can be equally effective in soliciting desired change and action. There is now evidence suggesting that more flexible, informal instruments might be equally, if not more effective in guiding States and other actor’s behavior, as long as the instrument is sufficiently precise and instructive, and has come about through a broad (multi-)stakeholder effort . On the It is also important to understand how new international legal norms may emerge – or even can be actively pursued, framed, diffused, put into effect, and ultimately implemented. Models that explain ‘international norm dynamics’ in social constructivist international relations theory provide useful insights into how key domestic and international actors, including so-called “norm entrepreneurs”, interact with each other in relevant global networks and organizations such as the WHO. They explain how a sufficient momentum for the adoption of new instruments may come about, or not , 21. In Finally, international relations theory suggests that any process of norm creation needs to start with agenda-setting and persuasion, or framing efforts by one or more dedicated “norm entrepreneurs” with (access to) sufficiently strong organizational platforms to spread their messages , 21. OneIn pursuing effective global law and policy responses to NCDs, it will be crucial not only to understand health science and policy options on NCDs and risk factors, but also to understand the various critical factors that determine the success or failure of global health law-making efforts. Such critical factors can relate to the presence of successful norm entrepreneurs, or to understanding better how non-binding WHO instruments can inform and strengthen domestic responses to NCDs, in particular, in comparison to binding instruments such as the FCTC.The role of law and policy in regulating modifiable behavioral risk factors, including tobacco use, alcohol, unhealthy diets and low physical activity, has gained increased attention in international and domestic NCD debates. This article highlights that there is considerable evidence for successful international and domestic legal interventions, but also identifies salient knowledge gaps on the exact role of and options for suitable law and policy responses. Therefore, and in order to meet the ambitious goals on NCD reduction now set out in the Sustainable Development Goals, this article identifies key questions for a bold interdisciplinary research agenda which requires researchers in various fields to work more closely together.


It has been a busy 5 years. So, on behalf of Emeritus Editor-in-chief Dr. Afzal Sheikh and my co-patron and Editor Dr. Bilal Mirza, let me take just a moment to celebrate and reflect. Five years may be the blink of an eye in the print publishing world, but it’s practically a lifetime online. At this juncture, we take time out to reflect briefly on what has been, and comment on future directions.As patrons and editors, I and Dr. Mirza had to work on few very vital issues. First and foremost concern was that it should not be looked upon an isolated Indo-Pak venture! Though in the first five years, 2/3rd of the contributions are from these two neighboring countries, the other 1/3rd valuable share comes from all parts of the globe . We have an international reach now. About the annual viewership statistics, the number of annual visitor have risen by 1/3rd, the pageviews have risen 250%. However, the percentage of returning visitors has not risen that substantially, only from to 16.9% [1] to 26.6% over 4 years. We have better exposure after being indexed with plethora of indexing agencies including PubMed and PubMed Central. On the first and second anniversaries of JNS, we had shared the top five manuscripts that had highest viewership in the year 2013 . On the fifth anniversary, we share here a simple and straightforward way to help gauge the productivity of a scholar. The following 6 manuscripts had more than 5 citations in international journals:We congratulate the team of authors from Niloufer Hospital, Institute of Child Health, Osmania Medical College, Hyderabad, Telangana, India whose case report has scored 12 citations till date; kudos for another addition to i10-index [4]. Journal of Neonatal Surgery has got 266 citations since its inception for 284 citable items. I am not unduly perturbed with the low 2-year impact factors of the journal for the last 3 years ; the reasons are rather obvious. One, globally the surgical journals usually have far lower impact factors as compared to basic or clinical medical research journals. Two, we do not represent any official journal of any Medical Association, thus keeping the politics of the academicians away. We shall strive to go higher solely on the strength of its diverse and high-quality content. We invite material on the latest innovations and developments that are happening in the developed world. If you value the benefits brought by publishing in a high quality open access journal, with a dedicated editorial team, and a rigorous but fair and friendly peer review service, we look forward to receiving your next submission to JNS.I close with once again expressing my sincere gratitude to our reviewers for their valuable time, as well as to our authors and the Journal’s Editorial Board for their most valuable support.Source of Support: NilConflict of Interest: None


Causes of “true” late kidney allograft failure remain unclear as study selection bias and limited follow-up risk incomplete representation of the spectrum.We evaluated all unselected graft failures from 2008–2014  by contemporary classification of indication biopsies “proximate” to failure, DSA assessment, clinical and biochemical data.The spectrum of graft failure changed markedly depending on the timing of allograft failure. Failures within the first year were most commonly attributed to technical failure, acute rejection (with T-cell mediated rejection [TCMR] dominating antibody-mediated rejection [ABMR]). Failures beyond a year were increasingly dominated by ABMR and ‘interstitial fibrosis with tubular atrophy’ without rejection, infection or recurrent disease (“IFTA”). Cases of IFTA associated with inflammation in non-scarred areas (compared with no inflammation or inflammation solely within scarred regions) were more commonly associated with episodes of prior rejection, late rejection and nonadherence, pointing to an alloimmune aetiology. Nonadherence and late rejection were common in ABMR and TCMR, particularly Acute Active ABMR. Acute Active ABMR and nonadherence were associated with younger age, faster functional decline, and less hyalinosis on biopsy. Chronic and Chronic Active ABMR were more commonly associated with Class II DSA. C1q-binding DSA, detected in 33% of ABMR episodes, were associated with shorter time to graft failure. Most non-biopsied patients were DSA-negative . Finally, twelve losses to recurrent disease were seen (16%).This data from an unselected population identifies IFTA alongside ABMR as a very important cause of true late graft failure, with nonadherence-associated TCMR as a phenomenon in some patients. It highlights clinical and immunological characteristics of ABMR subgroups, and should inform clinical practice and individualised patient care. RefinBut the question arises as to whether this is truly representative. Specifically, landmark studies from the Edmonton  and DeKAThe Mayo Clinic group  also idein the absence of prior or current specific diagnosis was more common, with 12/39 (31%) indication biopsies showing this lesion. The authors acknowledged that most (76.1%) grafts failing beyond 5 years did not undergo biopsy within 2 years of failure, and electron microscopy and DSA evaluation were unavailable. Nevertheless, other studies of non-selected graft failures beyond 3 months post-transplantation showed DSA at the time of graft failure in only 9/69 (13%) patients .de novo post transplantation (exact timing unknown). Subclassification of graft failures was as follows: Acute Active ABMR (n = 9); Chronic Active ABMR (n = 15); Chronic ABMR (n = 3). All except 4 patients displayed microvascular damage  as histological qualifiers; 3 patients with Chronic Active ABMR and 1 patient with chronic ABMR displayed macrovascular changes  without microvascular damage or inflammation.Twenty-seven grafts failed to ABMR. DSA was present at the time of biopsy, and persisted to the time of graft failure; all DSA developed Biopsies showing Acute Active ABMR displayed more glomerulitis and PTCitis than biopsies displaying Chronic Active ABMR . All bioThe characteristics of patients and DSA are also shown in Finally, ah-scores were lower in patients with Acute Active ABMR ; six disClass II DSA was more common in patients with Chronic Active or Chronic ABMR, compared with Acute Active ABMR . No diffNine patients (33% of patients with ABMR) displayed C1q-binding DSA , with no difference across histological subgroups. Mean time to graft failure was shorter in patients with C1q-binding DSA .Twenty-nine grafts failed with only IFTA on indication biopsy proximate to failure. None showed microcirculatory inflammation, vasculitis or C4d deposition; DSA was absent in all at the time of biopsy and at graft failure. Banff ci and ct scores were universally ≥2.By definition, no cases displayed interstitial infiltrates meeting acute rejection criteria (including borderline change). However, milder interstitial inflammation was seen in some biopsies, as was inflammation in scarred areas. We therefore segregated IFTA cases into 3 groups : IFTA winon-scarred areas had more likely experienced prior rejection , late rejection (>12 months post-transplantation), and nonadherence . No patients in other IFTA groups displayed nonadherence or late rejection. Time to graft failure was shorter, and renal function decline more rapid, in patients with inflammation in non-scarred areas compared with the other groups . Proteinuria at biopsy was numerically lower when inflammation in non-scarred areas was seen (p = 0.06). Interestingly, many patients with IFTA displayed marked proteinuria, but this was a manifestation of increasing glomerulosclerosis as part of the scarring process; proteinuria was highly correlated with the percentage of globally sclerosed glomeruli on biopsy in these patients with IFTA  glomerular diseases, which were considered as a separate category of graft failures.As defined, these groups displayed notable clinical differences. Specifically, patients displaying IFTA with inflammation in Percentage glomerulosclerosis, “cv” and “ah” scores were all lower in biopsies displaying IFTA with inflammation in non-scarred areas compared with the other 2 groups (p = 0.02 for both comparisons). The paucity of patients not treated with CNI precluded evaluation of the role of CNI exposure in the development of IFTA. More patients with graft failure classified as IFTA were treated with ciclosporin than tacrolimus, but this is confounded by the more recent introduction of tacrolimus into clinical practice coupled with the increased time post transplantation at which IFTA graft failures occurred.de novo) DSA (n = 2). However, these characteristics were mutually exclusive . The other 8 patients displayed TCMR without either DSA or microcirculation injury.TCMR was seen on indication biopsy in 14 patients. All cases met criteria for Banff 1B rejection. Histological features are shown in xclusive  and so nde novo) was detectable in 5 patients, with 16 demonstrating no DSA. These 2 groups did not differ numerically or statistically for any evaluated demographic or clinical factors, with slow progression to failure and moderate proteinuria at the time of failure  are shown in . Note thVery appropriately ABMR is considered a major cause of late graft failure. This study confirms its importance, and refines clinic-pathological correlations in the context of contemporary histological classification and antibody evaluation. Yet the attention to ABMR has been mirrored by the concept that other causes of graft failure, in particular progressive scarring (“IFTA”), are unimportant. The current study opposes this prevailing view by examining unselected patients with true late graft failure, which by virtue of design and duration of previous studies have been incompletely represented hitherto. Acknowledging this heterogeneity informs practice, and suggests individualisation of therapy directed towards the prevailing mechanism.improved graft outcome  within 5 years in the current study. Beyond 5 and 10 years post-transplantation failures became increasingly biased towards IFTA which therefore represented a dominant finding in “true” late graft failure. And it is precisely these failures at these late times which have proven so resistant to the advances and evolution of transplantation practice during the last 25 years [Advanced fibrosis (“IFTA”) in the absence of defined immune/inflammatory lesions was a major cause of graft failure in this series, particularly for late graft failures. Other studies suggest this lesion to be unimportant, although their design was such that IFTA was likely under-represented as discussed above –6. Even non-scarred areas, with these patients experiencing brisker functional decline. We contend that this biopsy finding may represent a low-grade alloimmune response, and attention to adherence and immunosuppression is mandatory.Inflammation was seen in some biopsies with IFTA, in the absence of other inflammatory diagnoses. This was either restricted to areas of scarring, or present in non-scarred areas but by definition not meeting criteria for rejection. Subgroup analysis revealed interesting clinico-pathological correlations. Specifically, nonadherence and late rejection episodes were unique to the group with inflammation in Yet the majority of patients with IFTA displayed either no inflammation or inflammation restricted to scarred areas. The clinical characteristics of patients with these findings were similar, without an abundance of rejection or nonadherence, and similar rate of graft decline. We therefore contend that inflammation within areas of transplant fibrosis may represent a nonspecific feature of the scarring process itself, which indeed is recognised elsewhere . Other sAn important question stemming from the current study is why IFTA may progress in the absence of inflammatory disease, a process previously coined “mysterious dysregulated fibrosis” ,17. ReceThe results of the current study require interpreting in the context of the changing face of transplant immunosuppression and donor/recipient demographics. The design of the study does not lend itself to interpretation of the influence of changes in clinical practice over time, and the results require careful interpretation in that context. Yet in parallel, a major strength is the examination of “true” late graft failure. It affirms the importance of ABMR in late graft loss, and highlights the importance of progressive fibrosis as important pathway to graft failure and target for intervention. This understanding of the true spectrum of graft failure is an important step in improving clinical practice by individualising patient care.


The current paper describes a rare case of a patient who suffered from unilateral apraxia of eye closure as a result of a bilateral stroke. Interestingly, the patient’s ability to voluntarily close both eyelids (i.e. blinking) was not affected, indicating that different neural mechanisms control each type of eye closure. The stroke caused damage to a large part of the right frontal cortex, including the motor cortex, pre-motor cortex and the frontal eye field (FEF). The lesion in the left hemisphere was restricted to the FEF. In order to further study the neural mechanisms of eye closure, we conducted an fMRI study in a group of neurological healthy subjects. We found that all areas of the oculomotor cortex were activated by both left and right winking, including the FEF, supplementary eye field (SEF), and posterior parietal cortex (PPC). Blinking activated FEF and SEF, but not PPC. Both FEF and PPC were significantly more active during winking than blinking. Together, these results provide evidence for a critical role of the FEF in voluntary unilateral eye closure.