JACMP Community! In search of inspiration for my inaugural editorial of my second round as Editor‐in‐Chief of the JACMP, I looked at some historical documents for guidance. The idea for an open‐access journal (that term did not exist at the time) was first discussed as I remember in November of 1997. Ann Wright had mentioned to me, in an almost offhanded way, that it would be very useful to have a standalone clinical medical physics journal, apart from Medical Physics. She was thinking of a print journal, but it occurred to me that the Internet (then only a few years old) would offer a great opportunity to publish a new type of journal — one that did not offer print issues, but would exist only online. The savings of printing and postage would allow for a new type of business model, one that could provide the articles at low or even at no cost to the author and reader.Greetings to the Journal for Clinical Physics. We had no concept of how difficult and complex it would be to realize this vision, or of the dearth of resources available to proceed. The following is the verbatim initial report of that task group as written in April of 1998. It shows, I believe, that we formulated the kernel of a vision that was later substantially realized, although we did get more than a few things wrong.Alex Turner, ACMP Chairman, thought that was an idea that should be explored and appointed a task group to investigate the possibility of a new JACMP became, I believe, the first open‐source and open‐access academic journal in Radiology and Radiation Oncology, and one of the first such journals in Medicine. Next issue, I will discuss how our vision for the JACMP has evolved over the years, and what we might expect for the future.In 2000, the Michael D. Mills, Ph.DEditor‐in‐ChiefOctober 8, 2012The idea that the medical physics community would be served by a clinical medical physics journal is not new. For some time, medical physicists involved primarily in clinical work have found it difficult to identify a forum which would allow for timely, wide, and archival distribution of clinical techniques and information. The advent of Web‐based technologies and the growing access of the medical physics community to the Web‐based information pool suggest an electronic journal is an idea worthy of consideration. The preliminary response of vendors suggests support for new and emerging avenues to distribute information respecting their products. Many vendors already maintain web sites and are looking to promote their use. Specific ideas to identify an editor‐in‐chief and an editorial board, establish and build the journal as a stand‐alone business, target participation from members of the larger medical physics community, develop a user profile for marketing purposes, and promote collateral growth within the ACMP are discussed. Some technical aspects respecting how the journal might operate are outlined. It is our hope to have a working demonstration of this journal and its proposed operation at the 1998 ACMP Board meeting.Medical Physics enjoys an international reputation for excellence among scientific journals. However, many valuable clinical papers which do not contain new science are rejected for review under existing editorial policies. The same can be said for Physics in Medicine and Biology, published in the UK by the Institute of Physics and Engineering in Medicine. Published by ASTRO, The International Journal of Radiation Oncology Biology Physics also enjoys worldwide respect. Physics articles are, however, limited and only a small percentage of medical physicists are members of ASTRO and receive the Journal. Medical Dosimetry is published by the American Association of Medical Dosimetrists. This Journal does not enjoy the reputation of the three journals first mentioned. In addition, few medical physicists are members of the AAMD and consequently receive the Journal. Proceedings published after clinical workshops are often of high quality; however, distribution is usually limited to participants in the conference. Task group reports address some clinical issues, but the scope of these reports is limited to the executive charge to the group. The investigation, drafting, and review process for these reports often takes years to complete. Proceedings, workshop reports, manuscripts rejected due to lack of space, and reports of individual projects practical in nature constitute a class of clinical information within the medical physics community which cannot find a timely, peer reviewed publication outlet which also reaches the general medical physics population.Published by the AAPM, The potential target audience is composed of all practicing clinical medical physicists, and a limited number of radiation oncologists, radiologists, dosimetrists, and administrators. Currently, we estimate approximately 70% of the target audience has access to the Web, with the vast majority of these having no better than 28.8 K modem communications. While the entire ACMP membership is part of the target audience, ACMP members comprise only 10% or less of the target audience.Medical Physics or Medical Dosimetry; 3) co‐publish articles with Administrative Radiology or Reports on Practical Oncology. Each of these avenues were found to have significant drawbacks respecting cost, vendor interest, current publication intent, scope of distribution, and potential of cooperation with other organizations. In particular, it was expressed the AAPM would not be comfortable with incorporating this type of activity into Medical Physics. Also, none of the above offer the advantages of an electronic journal discussed below.Suggestions include: 1) expand the ACMP, AAPM or IOMP newsletter to include peer‐reviewed clinical articles; 2) co‐publish clinical articles with Following is a suggested, but not exhaustive, list of the types of articles which might be published: 1) articles which describe a unique solution to a clinical problem for a specific patient, type of patient target volume, or piece of equipment; 2) equipment evaluation and review articles describing commissioning tests and techniques for clinical implementation; 3) articles which describe multiple types of equipment, often from different vendors, which must work together and communicate seamlessly; 4) health physics papers describing unique shielding or radiation protection problems; 5) papers dealing with Quality Assurance, Continuing Quality Improvement, and Total Quality Management; 6) articles dealing with administrative problems including staffing, space, equipment evaluation and purchase, project analysis, and resource planning; 7) review articles dealing with how different institutions and vendors approach similar clinical problems; 8) articles describing ways to implement and verify recommendations from scientific bodies  while recognizing specific equipment or clinical limitations; 9) business management articles addressing the changing reimbursement and management landscape and how individual medical physicists and the medical physics community might be proactive; 10) reviews of emerging technology, changing regulatory standards, and practice accreditation; 11) debates respecting clinical issues; 12) medical‐legal issues as they impact the practice of medical physics. The editorial board of the journal will be responsible to define the list of article types to be included in the journal.Medical Physics (Jan/Feb 1974) describes the purpose of that journal, but also suggests other legitimate professional interests: “…professional purpose(s)… deals with the individual personal interests of the professional person and might include legislative lobbying, wage and salary negotiations, and unionization. We have no taste for these …”An editorial by Gail Adams in Vol.1 No. 1 of Medical Physics since inception, but nevertheless are legitimate concerns of professional life. The articles published in clinical and professional proceedings associated with the ACMP meetings number between 20 and 30 per year. An initial proposed goal which we believe may be achieved is to publish 15 such papers per quarter, or 60 papers per year. It was agreed the journal should accept articles from anyone, at least initially.The proposed clinical medical physics journal would review and publish such articles which have been rejected by It is proposed the journal be published in quarterly issues with papers being posted to the current issue as soon as they are available. The journal might consist of seven sections: 1) Editorials; 2) Invited Papers; 3) Letters to the Editor; 4) Peer‐reviewed Articles 5) Peer‐reviewed Articles — sponsored by a specific vendor ; 6) Continuing Education Articles — perhaps associated with CAMPEP; and 7) Calendar of Meetings. Serious consideration was given to a suggestion that an employment notification service be included as part of the journal. This idea was rejected as potentially too controversial for this new venture. However, we recommend the ACMP Board revisit the need for the ACMP to operate and publish placement notices for its members. We propose that vendors be allowed to sponsor papers for inclusion in a designated section of the journal. Sponsorship buys a one‐page advertisement to be included at the end of the paper and printed out upon retrieval. Also, the individual will receive a link to transfer to the web site of the vendor to view the latest offerings. This feature will exist in perpetuity for this paper at an initial cost of $500.00 to the vendor. This cost might be adjusted on a sliding scale according to the length of the article. These features allow the vendor to publish articles presenting technical and clinical specifics of a product, and to present timely information specific to the range of products offered by the vendor. Also, vendors will be listed along a bar on the left side of the journal page and vendor ads will flash every 20–30 seconds at the top or bottom of the page. This benefit to the vendor can only be accomplished using an electronic medium.Vendors are going to support the journal on the basis of the number of “hits” or different individuals visiting the site and seeing the flashing ads each month. A good number in the industry is $0.25 per pair of eyes per month, or $0.75 per pair of eyes per quarterly issue, or $3.00 per medical physicist per year. A reasonable goal is 1000 “hits” per month from medical physicists. A vendor may be persuaded on this basis to support the concept of a journal during start‐up at a rate of $2,000.00–$4,000.00 per year, provided we demonstrate this level of activity. To attract younger medical physicists to the journal, continuing education features would be a prominent focus of the journal's mission.The advantages of an electronic journal include:1) lower operating and distribution costs; 2) worldwide access to the medium; 3) searching capabilities; 4) potential for multimedia capabilities including sound and moving pictures; 5) for those who do not subscribe, the potential for charging for articles on a flat‐fee per download basis; 6) there are some specific services which may be offered to supporting vendors which are only available if the format is electronic.Articles will be submitted in Word‐6 or WordPerfect format with imbedded Excel, JPG or GIF files for tables and figures, receptively. They will pass through the entire review process in this manner. Once accepted for publication, the articles will be written by Adobe Acrobat into PDF format. They may be viewed online and in color, as they will appear on paper using this format. Adobe Acrobat Viewer is free software and is being distributed with the AAPM CD‐ROM to all AAPM members this month. It operates as a plug‐in for both the Netscape and Microsoft Web browsers. The articles may be downloaded and printed out by anyone who has access to the Web.The journal could operate in association with the ACMP Web page, which currently exists on the ACR server as part of the ACMP contract. Routine backup procedures and password protection are already in place for this server.Medical Physics, PMB, and / or IJROBP. The level of support for this position should be equivalent to that offered to the program of the current editor of Medical Physics, or $35,000.00 per year. This figure is based on six hours per week support for the editor and four hours per week support for a secretary. The editor‐in‐chief will be responsible for recruiting associate editors and peer reviewers, and determining the content of each issue. He/she will be responsible for assigning a retention period associated with the issues and articles before archiving to CD‐ROM or equivalent. The Managing Editor will be responsible for the operation of the journal and establishing vendor support. The Webmaster will be responsible for Web administration of the journal, including detailed standards for submission of articles. The financial matters for the journal would be handled by the Journal Finance Committee appointed by the ACMP Chairman during the three‐year start‐up period. These positions will be unpaid during the three‐year start‐up period. Total initial budget for this venture is $35,000.00 for support of the Editor‐in‐Chief, $5,000.00 for publicity, $5,000.00 for an assistant to the Webmaster, and $10,000.00 contingency fund to be used at the discretion of the Editor‐in‐Chief; total $55,000.00.Journal operation will be under the director of an editor‐in‐chief and a managing editor. The editor‐in‐chief should be an ACMP member who enjoys universal respect within the medical physics community. Ideally, he/she should be a past chair of the ACMP, a past president of the AAPM, winner of national and international honors such as Williams and/or Coolidge awards, and demonstrate a long history of editorial involvement with Medical Physics is $175.00 per year for individuals and $550.00 per year for libraries. However, any nonsubscriber may obtain a limited password by answering the questionnaire above; this individual will still be able to: 1) view, search and download the first three years of the journal; 2) view and search all abstracts for all years; and 3) download a specific paper for a user fee paid by credit card for a suggested price of $20.00. In comparison, to order a reprint from Medical Physics online costs $12.95 for shipping and handling and $15.00 royalty fee; total $27.95.Journal access may be password protected, but the passwords could be offered for free during the start‐up period. Individuals may obtain a password online by answering a short questionnaire about their education and employment background. This information will be used to identify a user profile so vendors may be persuaded to invest money in the concept. After the three‐year start‐up period is complete, total access for download will be restricted to ACMP members or subscribers only, utilizing assigned password access. Subscription cost should be similar to the cost of ACMP membership for one year; current subscription cost for The free password to the journal marketing should be limited to all medical physicists within the United States. Notice of the existence of the journal may be sent online to medical physicists with email addresses. A one‐page mailing should be prepared and placed in the ACMP and AAPM mailings for fall of this year. The mailings should be an announcement of the existence of the journal and an invitation to submit papers. The AAPM mailing will cost the ACMP $300.00 insertion fee plus the cost of producing and shipping 4,500 fliers to the AAPM headquarters. This mailing should be performed three times. The ACMP could purchase the membership list from AAPM and perform a separate mailing. A second and third mailing will probably be required to expose fully the journal to the medical physics community. Hard copies of the journal should be placed at the ACMP booth during meetings, along with a free download service for medical physicists.American Clinical Medical Physics Journal (ACMPJ) may be the most cost‐effective and realistic vehicle to accomplish this aim. Although the cost of the operation during the start‐up period is significant , the cost of walking away from this opportunity may be even greater. We hope to provide evidence of significant vendor support for this concept at the ACMP Board Meeting, as well as a working sample journal page.The key to success of any company, organization or institution is the management of information, which is the lifeblood of its existence. Individuals and professionals cannot compete and survive in a hostile environment unless they have ready access to the information which allows them to survive in such an environment. The challenge to the ACMP at this point in time is either to grow or die. To provide the medical physics community with essential, timely medical physics professional and clinical information may be our best chance to double in size within the next 3–5 years. The


Modified starches exhibited altered granule morphology and size compared to the control. About 20–30% of the transgenic lines of each series showed red-staining granules upon incubation with iodine, and contained higher phosphate content than the blue-stained starch granules. Moreover, low amylose content and altered gelatinization properties were observed in these red-stained starches. Principle component and correlation analysis disclosed a complex correlation between starch composition and starch physico-chemical properties. Ultimately, the expression level of endogenous genes involved in starch metabolism was analysed, revealing a compensatory response to the decrease of phosphate content in potato starch. This study provides a new perspective for engineering starch phosphate content in planta by making use of the compensatory mechanism in the plant itself.Phosphate esters are responsible for valuable and unique functionalities of starch for industrial applications. Also in the cell phosphate esters play a role in starch metabolism, which so far has not been well characterized in storage starch. Laforin, a human enzyme composed of a carbohydrate-binding module and a dual-specificity phosphatase domain, is involved in the dephosphorylation of glycogen. To modify phosphate content and better understand starch (de)phosphorylation in storage starch, laforin was engineered and introduced into potato . Interestingly, expression of an (engineered) laforin in potato resulted in significantly higher phosphate content of starch, and this result was confirmed in amylose-free potato genetic background ( Arabidopsis have indicated that phosphoglucan phosphatase starch excess 4 (SEX4) cooperates with like-SEX4 1 and 2 (LSF1 and LSF2) proteins to remove phosphate groups . Absorbance at 750 nm was then measured in a Model 680 XR Microplate Reader , and a calibration curve was used to calculate the concentration in nmol PO4/mg starch.Total phosphate content in starch was determined essentially according to the method of Morrison  with somAll analyses conducted in this manuscript have been performed in duplicate unless indicated otherwise.2/KI) and were then investigated with light microscopy . Further, scanning electron microscopy (SEM) Phenom™  was used to examine the detailed morphology on the granule surface. Starch samples of approximately 1 mg were evenly distributed on a carbon tabs  and mounted onto 12.70 mm aluminium specimen stubs , followed by coating with gold using a sputter coater . All images were digitally recorded.Starch samples were stained with 20×diluted Lugol’s solution (1% Io), peak (Tp) temperature of gelatinization and the melting enthalpy  were determined using Differential Scanning Calorimetry (DSC).Particle size distribution and gelatinization properties of starches were analysed as described in Ji, Vincken (35). The onset  was used to determine amylopectin chain length distribution. Degree of polymerisation (DP) 6–35 was separated according to the method of Huang, Nazarian-Firouzabadi .Starch moisture content was measured in duplicate using standard methods (AACC Approved Method 44–15.02), which involved oven drying at 105°C for 16 hours.Starch content was determined as described by Kok-Jacon, Vincken, Suurs and Visser .EF1α [6 and then converted to log 10. Ultimately, the resulting value (v) was used to assign transformants to different categories: none , low , medium  and high  expressers.Expression levels of (engineered) laforin from all transformants were determined in triplicate by qRT-PCR using the primers listed in EF1α . After nSEX4 [LSF1 and LSF2 [SP [AMY23 and β-amylase genes BAM1 and BAM9 [ISA1, ISA2 and ISA3 [SBEI and SBEII [SSSII and SSSIII [Ef1α gene as a reference gene, employing the 2-ΔΔCT method.Five randomly-selected transgenic lines from D, CD and SD series were used for gene expression analysis of key genes involved in the starch metabolism. In total, four starch-degrading genes and seven starch-synthesizing genes were selected according to previous studies, including phosphoglucan phosphatase starch excess 4 gene SEX4 , like-SEand LSF2 , 45, staLSF2 [SP , α-amylaand BAM9 , 48, isoand ISA3 , 49, stand SBEII , 51, sold SSSIII , 52. TheSignificant differences between modified starches and control samples in phosphate content and granule size were assessed by one-way analysis of variance (ANOVA). The least significant difference values were calculated at 5% probability. Statistical significances between gene expression level of transformants and that of the control were determined by using t-test. Inter-relationships between starch components and starch properties were analysed by means of Pearson correlations. All statistical analyses were performed using GenStat (16th edition). Principle component (PCA) analysis was performed using PAST software Package .In total, three transgenic series containing, respectively, DSP (D), CBM20-DSP (CD) and SBD-DSP (SD) were generated by introducing three constructs into potato plants. About thirty independent plants from each transgenic series and the control were multiplied and grown in the greenhouse. Plants were monitored in all stages and no significant differences were observed in plant architecture, tuber morphology nor tuber yield relative to the control plants (data not shown).Expression of SD in transformants was examined by qRT-PCR and Western dot blot analysis. According to gene expression level, transformants were classified into four different categories: none (N), low (L), medium (M) and high (H) expressors. All SD lines showed gene expression, and 28%, 34% and 38% of transformants were categorized as L, M and H expressors, respectively . FurtherLight microscopy (LM) and scanning electron microscopy (SEM) were used to investigate how the accumulation of the (engineered) laforin protein affects the starch granule morphology. About 22% of the D transformants, 19% of the CD transformants and 27% of the SD transformants showed red granules with blue cores of varying size when stained with iodine . FurtherIn addition, the exhibition of red-stained granules strongly suggests the reduction of amylose content in these starch granules, therefore, the amylose/amylopectin ratio was determined in all modified starches and the apparent amylose content (AM%) was calculated . The resGBSSI in transformants, as GBSSI is responsible for the amylose synthesis. Hence, GBSSI expression was investigated for transformants from each series, containing transformants with eight red-stained starches and seven blue-stained starches in total. The results revealed that amylose content strongly correlates with relative expression of GBSSI . Within each series, the red-stained group had a higher phosphate content than the blue-stained group. To illustrate, the phosphate content of the red-stained group from D series was ~32% higher than that of the control, and ~9% and ~6% higher than the red-stained group from CD and SD series, respectively. In each of the three blue-stained groups, the starch phosphate content was ~11% higher than that of the control, while between the three series of transformants no significant differences were observed.As shown in amf) and generated ~30 transformants of each transgenic series . Starch phosphate content was measured for each line. Results showed that the phosphate content in amfD and amfCD starches were 26% and 12% higher than that in control UT-amf, respectively (amf background provides confidence that starch phosphate content is affected by the (engineered) laforin, and not only by GBSSI suppression.In the red-stained starches the increase in phosphate content appeared to be linked to the increase in amylopectin content rather than to the expressed laforin. However, blue-stained starches containing unaltered amylose content also showed a significant increase in phosphate content, which indicated that the (engineered) laforin plays a role to increase starch phosphate content. To further investigate this, the same constructs were transferred into amylose-free potato mutant . By contrast, starches from the red-stained group in D series did not show any significant difference in granule size. Further analyses showed that there was no significant correlation between the expression of the (engineered) laforin and granule morphology or granule size.The median granule size (d50) was determined for all starches by analysing granule size distribution. The transformants with CD and SD proteins had larger starch granule size compared to the control . In parto and Tp (H) of these red-stained starches was consistently lower than that of the control. Unlike the red-stained starches, the gelatinization characteristics did not differ between the control and any of the blue-stained starches.DSC analysis disclosed that red-stained starches had a significantly higher To and Tp , suggestStarches, after debranching with isoamylase, were analysed with HPAEC (chain length ranged from DP6 to DP35). No significant changes in amylopectin fine structure were observed relative to the control (data not shown). Similarly, starch content was measured for the selected starches and no consistent changes were found compared to the control .o, Tp and Tc). The modified starches with relatively higher phosphate content, circled by the dotted line, had higher gelatinization temperatures, which is consistent with the visual inspection of the data in H) and granule median size (d50). D series (red dots) was separated from CD and SD series , while CD and SD modified starches did not separate from each other, suggesting that DSP functioned differently when alone than when appended to a glucan-binding domain (CD and SD). The analysis also showed that starches from CD and SD series had larger granules and higher MC than that from D series.To divulge more information resulting from combined effects of the different starch characteristics, a principal component analysis (PCA) was performed for all samples. As shown in r = 0.6 or 0.7) with gelatinization temperatures , and negatively correlated with gelatinization enthalpy  and amylose content . In contrast, AM displayed remarkably opposite correlation patterns with P, exhibiting a negative association to To, Tp and Tc (r = -0.8 or -0.6), and a positive correlation to ΔH (r = 0.7). Negative correlations between starch moisture content (MC) and gelatinization temperatures (r = -0.4 or -0.6) were observed. Furthermore, starch granule median size (d50) showed a negative correlation (r = -0.2 or -0.3) with To, Tp and Tc, but its associations with starch compositional characters  were not significant.Correlation analyses were performed to dissect the underlying relationships between starch compositional characteristics and starch physico-chemical properties . PhosphaGBSSI expression. Interestingly, in general, the results displayed increased expression of starch dikinases, GWD1 and GWD3, in all selected transformants compared to the control  between the expression levels of both genes was found . For the AMY23 expression, an increase up to 20-fold (p < 0.05) was found in transfomants from CD series compared with the control, but the increase in transformants from D and SD series was smaller. For other starch-synthesizing genes no consistent differences between transformants and the control were detected for any of the series (SP expression and GWD1 expression (r = 0.9), as well as GWD3 expression (r = 0.8), indicating that these three genes are co-expressed . This finding seems to conflict at first glance with the results of Worby, Gentry [in vitro. Clearly, the environment in potato amyloplasts is much more intricate than that in the in vitro system, and one important difference is the presence of other endogenous genes, which may be an obstacle to detecting the individual effect of laforin expression on phosphate content in potato. Therefore, we investigated whether the introduction of laforin affects the expression of other genes involved in starch phosphorylation pathway. Interestingly, the expression levels of starch phosphorylating genes, GWD1 and GWD3, were significantly increased in the different transformants. Moreover, although modest, a positive correlation was found between starch phosphate content and the transcript levels of both GWD1 and GWD3 genes  laforin in potato induces starch phosphorylation via GWD1 and GWD3 and thereby leads to an increase in phosphate content in starch granules, albeit the exact mechanism by which this would work remains to be elucidated and study on protein level is needed to further confirm the hypothesis. It has been demonstrated that starch phosphate groups are essential for well-functioning starch metabolism and plant performance  laforin activity and GWD1/GWD3 activity, and each of which is difficult to quantify. A similar compensatory regulation was also proposed by Mahlow, Hejazi, Kuhnert, Garz, Brust, Baumann [Arabidopsis GWD1-deficiency mutants.In this study, laforin, a human phosphatase gene, was engineered and introduced into potato plants to modulate phosphate content in starch granules, thus exploring the effect of phosphate content on starch properties and the mechanism of phosphoglucan metabolism in storage starch. When setting up the experiments, the idea was to generate starches with low amounts of phosphate content by expressing the (engineered) laforin gene in potato tubers. However, contrary to the expectation, the phosphate content increased substantially in starches from the transgenic lines compared with that of the control plants regardless of the construct and genetic background . It is  Baumann , who detOne other clear effect of the transformation was that about 23% of modified starches displayed red-stained color and low amylose content, and further analysis revealed that the lower content of amylose was caused by GBSSI suppression . MoreoveGBSSI gene, which would also explain the results presented here, as the promoter and transit peptide used in the constructs were derived from the GBSSI gene. However, since in this study the occurrence rate of modified starches with low amylose content is dramatically higher than the above-mentioned studies, it suggests that the upregulation of GBSSI might also be a consequence of the presence of the (engineered) laforin.Notably, GBSSI suppression in transformants has been observed in earlier studies using similar constructs containing GBSSI promotor and transit peptide, however, this phenomenon occurred in a much lower ratio, about 3% , 58. TheSP and that of phosphorylating genes, GWD1 and GWD3  laforin on genes involved in starch biosynthesis/degradation rather than due to the direct activity of the (engineered) laforin enzyme. This might be the reason why no significant correlation has been found between laforin gene expression and starch granule morphology, size and properties. The CD and SD starch granules showed a larger size than the control, which is in accord with previous observations that the introduction of starch-binding proteins in potato could alter the granule morphology and granule size , 36, 62.r = -0.7) was found between phosphate content and amylose content , which is supported by previous studies [Correlations between different starch compositional characters and starch properties have been identified. A strongly negative correlation  in potato starch is an efficient way to modulate starch phosphate content in storage starch. Intriguingly, this study reveals that potato tuber starch, containing by nature already high level of starch phosphate, has obviously a strong compensatory mechanism to modulate starch phosphate content during starch biosynthesis. A better understanding of this compensatory mechanism may provide better insights on how to modify starch phosphate content in potato starch.S1 FigBacillus circulans, an artificial PT-linker, ampicillin resistant gene and terminator, respectively. To generate this construct, a sequence encoding the potato GBSSI promoter and part of GBSSI transit peptide (HindIII—HindIII) was amplified from the construct pBIN19/SBD2 [AAGCTTAATACTAAAAAATGCAACAAAAT-3’5’-CC and AAGCTTGTTAACAGCCCTTAAACCAT-3’5’-CC and inserted into the corresponding sites of the pUC19 vector. The orientation of the HindIII—HindIII fragment was verified by sequencing. Subsequently, the sequence containing a SBD2 fragment and a part of GBSSI transit peptide (HpaI—BamHI) was amplified from pBIN19/SBD2 with primers GTTAACAAGCTTGATGGGCTCCAATCAAGAACT-3’5’- C and GGATCCGCCAAAACAGCCAAGCTTATG-3’5’-CG, followed by cloning into corresponding sites of pUC19.SBD, Linker, Amp and Ter stand for starch binding domain of cyclodextrin glycosyltransferase from N19/SBD2  with pri(TIF)Click here for additional data file.S2 FigSEX4), like-SEX4 genes (LSF1 and LSF2), starch-branching genes (SBEI and SBEII) and isoamylase genes , soluble starch synthase genes (SSSII and SSSIII) and β-amylase 1 (BAM1). The qRT-PCR was performed on control tubers (UT) and five random-selected transgenic tubers from each series, containing transformants with eight red-stained starches and seven blue-stained starches. The values are expressed as the mean ± S.D. from three independent measurements. No consistent changes in the expression level of these genes were observed relative to the control.Include: phosphoglucan phosphatase starch excess 4 ((TIF)Click here for additional data file.S1 Table(PDF)Click here for additional data file.S2 Table(PDF)Click here for additional data file.


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.Alexandre HuebAna Maria SilvaAnthony PanosCelso Salgado de MeloChiara GattoEdmo Atique GabrielEduardo Keller SaadiFabio GaiottoFábio TaniguchiFernando AntonialiGabriel LiguoriGilberto Venossi BarbosaJoão de Deus e BritoJosé Carlos PachónLindemberg SilveiraLuís Alberto Oliveira DallanLuiz Antônio Castilho TenoLuiz Antônio RivettiMarcos Aurélio Barboza de OliveiraMaurício MachadoMelchior LimaMichel Pompeu Barros de Oliveira SáNicolas BrozziPaulo Roberto B. ÉvoraReinaldo BestettiRicardo DiasRicardo de Carvalho LimaTania Strabelli


The American Psychosomatic Society is one of the oldest and probably the most influential scientific society in psychosomatic/biopsychosocial research worldwide. The current article delineates the historical development and current strategic orientation of the society.Review of published literature, archived materials and current documents of the society.Psychosomatic Medicine, which had already been founded in 1939 and has become one of the major journals in the field. As an organization, APS has developed into a premier international scientific society, providing an interdisciplinary home for researchers from medicine, psychology and related areas, gathering under the mission “to advance and integrate the scientific study of biological, psychological, behavioral and social factors in health and disease” and dedicated to the goals of Scientific Excellence, Clinical Relevance, and Vibrant and Diverse Membership. Besides editing Psychosomatic Medicine, the APS organizes Annual Meetings and specialized events, issues several scientific awards and scholarships and is engaged in collaborative efforts to improve the research and funding landscape for biobehavioral research in the US and translate psychosomatic research findings into medical education and clinical practice.The American Psychosomatic Society (APS) was founded in 1942, originally named the “American Society for Research in Psychosomatic Problems “. It originated from the editorial board of the Journal th anniversary year, the American Psychosomatic Society has developed into the scientific landscape of the 21st century, and its current updated strategy addresses contemporary demands in advancing science and improving holistic patient care.In its 75 The origin of Psychosomatic Medicine in the United States of America is often attributed to Helen Flanders Dunbar of New York and her publication in 1935 of “Emotions and Bodily Changes, A Survey of Literature on Psychosomatic Interrelationships 1910-1933” )ce paper , where Ece paper ; p. 134)ce paper  Based once paper .Psychosomatic Medicine and that “many more general scientific and medical journals regularly accept papers which are basically psychosomatic than was the case in earlier years.” The APS membership had grown to 950 by 1984. However, Jenkins stated that psychosomatic research still paid insufficient attention to individuals’ environments, cultural factors and to persons in childhood, old age or minority populations, as well as to treatments and rehabilitation for major illnesses, which he considered desirable for the future.As time progressed, Jenkins  found psA membership survey conducted in 1987  found thth anniversary was celebrated with a large international meeting held in New York in 1992. At that time, the Society had, in 1986, appointed a professional manager and was financially healthy due to a new publisher and contract for the Journal Psychosomatic Medicine which generated substantially increased and reliable royalties. These changes gave the Society the opportunity to engage more in committee work and press relations. Awards and scholarship programs were created and the Society started a newsletter as one of several measures to keep members more actively involved [APS’ 50Psychosomatic Medicine. One important initiative in this context focused on advancing neuroscience in psychosomatic research by organizing dedicated symposia at APS meetings, reviewing relevant literature in several articles in Psychosomatic Medicine and encouraging original research by, eg, the donation of a dedicated annual award [However, despite several efforts to keep the balance between psychosomatically-oriented internists or other medical specialists on one hand and consultation-liaison psychiatrists on the other, and between clinicians and more basic researchers, the clinical focus in APS diminished further in the following two-and-a-half decades. The position of Ph.D.s in the Society was strengthened in order to meet the interests of health psychologists and basic researchers. This undoubtedly led to a strong research record of the APS membership whose publications now appear in highly ranked medical and psychological journals, in addition to the continuously high quality of research published in al award .M.D. members lost their majority in the Society in 2001 and never again reached a portion of >50% of APS members. Since 2009 M.D.s make up some relatively stable 30% of the membership. The decline in the numbers of (often older) M.D. members went along with an increase in younger members (<47 y/o) from 25% in 2000 to a maximum of 46% in 2014 and a related increase in members holding neither an M.D. nor Ph.D. degree from 10% in 2000 to approximately 20% in recent years (unpublished data). Over the last 20 years, APS Presidents have been alternating at approximately equal rates between M.D.s and Ph.D.s [In contrast to this development in APS, institutionalized psychosomatic medicine gained increased credit by the creation of a subspecialty in “Psychosomatic Medicine” for psychiatrists in 2005, mainly reflecting consultation-liaison psychiatry. However, the new specialty found little resonance in the APS and most clinical psychiatrists specialized in “Psychosomatic Medicine” are now organized in the Academy of Psychosomatic Medicine, while the majority of APS members is nowadays made up by non-physician researches.The decreasing number of M.D. members and the parallel increase in basic “mechanistic” research  almost As early as in 1960, Wittkower  had statPsychosomatic Medicine”. After several months of discussion the committee decided to keep the name of the journal and also not to add anything to it. However, with its 70th anniversary in 2009, the editors of Psychosomatic Medicine – with the approval by the APS Council – finally added the subtitle “Journal of Biobehavioral Medicine” to the (unchanged) name of the journal. When the subtitle was added to the journal, the editors stated that it aimed at helping the journal to reach out to the broader medical community and to reconcile “the journal’s identity with its 21st century content.” [In 1977, an ad-hoc committee was created to consider a name change or a subtitle for the journal “ontent.” . Howeverontent.” .This occurred during a phase of discussions within the membership of APS whether it should not better change its name in total. Several members had several reasons for considering a name change. Its partly negative connotation among somatic clinicians and the lay public, who often use it in a stigmatizing way, was one key argument. Unlike other countries such as Germany or Japan, the term “psychosomatic” has negative connotations among the lay public and many health practitioners in the United States, where it is often equated with imaginary symptoms rather than (measurable) mind-body interactions. In addition, the term was thought to perpetuate the mind-body dualism it originally intended to overcome by, eg, ignoring the social dimension. Finally, many behaviorally or biologically oriented researchers perceived “psychosomatic” as too closely linked to “psychoanalytic”. In fact, psychoanalytic psychiatry while having been at the heart of APS in its early years has been the theoretical and clinical background of only a small minority of members in recent years. A majority of members now understand themselves as behavioral, biological or “biobehavioral” researchers, as epidemiologists, neuroscientists or physician scientists. The name change discussions started in the late 1990s and several alternative names were brought up during the process. However, none of the alternative names developed sufficient traction to justify the abandonment of history and the costs associated with rebranding.The discussion only came to an end after a membership vote in 2013, where members were finally invited to decide whether they wanted to keep the historical name or rename the society to “Association for Biopsychosocial Medicine”. Although “biopsychosocial” was widely acknowledged to cover the interpersonal and social aspects of psychosomatic medicine better than the original term, a majority finally voted in favor of keeping the original name . This doth anniversary in 2017/18. However, APS has, in early 2016, refined its goals, adapted a new mission and strategic goals and launched a number of activities to achieve those goals [So APS is still the American Psychosomatic Society when it comes to celebrating its 75se goals , 25.www.psychosomatic.org) now reads: “To advance and integrate the scientific study of biological, psychological, behavioral and social factors in health and disease.”The mission statement as published on the APS website , aiming at integrating efforts across organizations to clarify major biobehavioral research questions and guiding scientific directions to advance and coordinate clinically relevant science in biobehavioral medicine.Collaborate with the Consortium of Social Science Associations (COSSA ), providStrategies related to the goal of scientific excellence include:Education: APS is working with authors of educational videos and other materials for the APS website to translate research findings into clinical application. APS members are also working on a comprehensive educational textbook on psychosomatic medicine and on educational programming during Annuals MeetingsAPS is liaising with the Integrated Primary Care Alliance(IPCA) to improve biopsychosocial understanding and practice in primary care settingsPsychosomatic MedicineA dedicated Clinical Series is being prepared for APS started cross-advertizing for events with the Academy of Psychosomatic Medicine as a mainly clinically-oriented partner society.The following strategies are related to the goal of clinical relevance:Several ongoing scholar and travel awards mainly targeting trainees and junior scientists from the US and worldwideA new international lab-to-lab exchange program  for whicThe anniversary awards mentioned above, which offer an opportunity for senior and mid-career researcher to get their outstanding work recognized beyond the existing APS awards such as the Herbert Weiner Early Career Award, the MacLean Award for outstanding neuroscience research in Psychosomatic Medicine, the Patricia A. Barchas Award for Sociophysiology, the Distinguished Scientist Award and the Donald Oken Fellowship honoring a distinguished consultation-liaison psychiatrist or internist who hasn’t previously attended an APS meeting.As related to the goal of fostering a vibrant and diverse membership the following initiatives are underway:th anniversary [st, 2017 and culminate in a series of events during the 2018 Annual Meeting, which will take place in Louisville, Kentucky on March 7–10, 2018. For the anniversary celebrations, APS is currently conducting some research into its history, taking advantage of a rich pool of archived materials ranging back to the very beginnings of the society, collecting historical memories by interviews with senior members and addressing historical aspects during the jubilee meetings. Readers are cordially invited to submit historical materials to the APS national office at info@psychosomatic.org and to join us in celebrating the 75th anniversary and developing American and international psychosomatic medicine into the future.This year, APS is celebrating its 75iversary , startin


Therefore, in the background of the double mutant dpe2/phs1, a triple mutant was generated lacking the initial starch degrading enzyme glucan, water dikinase (GWD). The triple mutant showed improved plant growth, a starch-excess phenotype, and a homogeneous starch distribution. Furthermore, the number of starch granules per chloroplast was increased and was similar to wild type. However, starch granule morphology was only slightly affected by the lack of GWD as in the triple mutant and, like in dpe2/phs1, more spherical starch granules were observed. The characterized triple mutant was discussed in the context of the generation of starch granules and the formation of starch granule morphology.An Arabidopsis double knock-out mutant lacking cytosolic disproportionating enzyme 2 (DPE2) and the plastidial phosphorylase (PHS1) revealed a dwarf-growth phenotype, reduced starch content, an uneven distribution of starch within the plant rosette, and a reduced number of starch granules per chloroplast under standard growth conditions. In contrast, the wild type contained 5–7 starch granules per chloroplast. Mature and old leaves of the double mutant were essentially starch free and showed plastidial disintegration. Several analyses revealed that the number of starch granules per chloroplast was affected by the dark phase. So far, it was unclear if it was the dark phase  Ho. Ho8]. HMaltose exporter 1; , a, adpe2-1; sex1-8 . The gendpe2-5, phs1b, sex1-8, and dpe2-5/phs1b in the Col-0 background have been described previously , ), a), adpe2/c enzyme  should bphs1b exhibited the typical flat discoid Arabidopsis starch granules. Dpe2 and dpe2-5/phs1b showed rounder starch granules, as described , wh, whphs1bdetected . Furtherobserved .dpe2-5/phs1b (dpe2-5/phs1b and dpe2-5/phs1b/sex1-8 revealed only small differences (dpe2-5/phs1b, whereas in the other mutants it has a dominant effect. However, the lack of GWD has an important effect on the starch granule size. Thus, for all mutants lacking GWD, the starch size is massively increased to 5–6.5 μm (Thus, the starch morphology is massively affected by the lack of GWD in these two mutants. In contrast, the starch granules isolated from the triple mutant revealed no obvious differences in their morphologies compared to -5/phs1b . Similarferences . Thus, G5–6.5 μm . However5–6.5 μm  was acco5–6.5 μm .dpe2-5/phs1b/sex1-8, we were able to show that the reduced number of starch granules per chloroplast in dpe2/phs1 was dependent on ongoing starch degradation in the dark, whereas the starch morphology, and thus the formation of more spherical starch granules, was only slightly influenced by early inhibition of starch degradation.In summary, by generating


We present a 7-year-old patient who was diagnosed with asymptomatic nephropathic cystinosis following the detection of the pathognomonic corneal white crystalline opacities during a routine eye examination. The nephropathic form is further divided into two subtypes: infantile cystinosis and juvenile cystinosis.4 The non-nephropathic form is also known as ocular cystinosis. In ocular cystinosis, pathognomonic yellow-white crystalline deposits are observed in the cornea, but the kidneys are not affected.4Cystinosis is a rare metabolic disease with autosomal recessive inheritance,A 7-year-old female patient with no complaints presented to our clinic for routine eye examination. Her uncorrected vision was perfect in both eyes. On anterior segment examination, yellowish-white crystallized opacities were observed throughout the corneal stroma bilaterally , 1B. Ant4 Electron microscopic and ultrastructural examinations of the crystals have shown that they formed of intralysosomal L-cysteine.5 Most of the cases in the literature are of infantile type, which is the clinically most severe form.6Cystinosis is a lysosomal storage disease with an autosomal recessive inheritance pattern. Impairment of the transporter system responsible for transporting cystine out of lysosomes results in the accumulation of cystine crystals in tissues such as the kidneys, eyes, bone marrow, liver, spleen, pancreas, thyroid, skeletal muscles, thyroid, and brain.6 These deposits can be located in the corneal epithelium, stroma, and endothelium, and their distribution may be associated with disease course and duration.7 The corneal crystals begin to accumulate in infancy, being found in almost all cases of nephropathic cystinosis at 16 months.7 They first accumulate in the peripheral cornea and progress toward the center with age.8 While these crystalline deposits often do not cause visual impairment, in rare cases they may form band keratopathy, thus affecting the central cornea and reducing visual acuity.9 Superficial punctate and filamentary keratopathy are frequently observed in adults, whereas band keratopathy, peripheral corneal neovascularization, and posterior synechiae associated with increased iris thickness are seen in the elderly.10 Rare long-term complications of infantile and juvenile cystinosis include patchy pigmentary retinopathy, pigmentary changes in the macula, and involvement of the iris, ciliary body, choroid, and optic nerve due to the cystine accumulations.10 The most common symptoms in older patients is photophobia and associated blepharospasm.11 In addition, color vision, peripheral vision, and night vision may be reduced due to anterior and posterior segment complications.11 Depigmentation of the peripheral retina with pigment epithelium mottling is the most common posterior segment complication.12 In about 10-15% of patients, retinopathy leads to blindness.13Ocular cystinosis is diagnosed when the pathognomonic refractive cystine crystal accumulations are observed throughout the conjunctiva and entire cornea  on anterior segment examination.de novo, recent refractive or corneal surgeries, and interventions such as keratoplasty. Corneal dystrophies that cause crystalline keratopathy include Schnyder crystalline cornea dystrophy and Bietti crystalline corneoretinal dystrophy. Schnyder dystrophy is a slowly progressive corneal dystrophy with autosomal dominant inheritance.14 Clinical manifestations include opacification of the central or midperipheral cornea, dense arcus senilis, reduced corneal sensitivity, and recurrent corneal erosion.15 Bietti corneal dystrophy shows an autosomal recessive inheritance pattern and is characterized by progressive night blindness and narrowing of the visual field. Clinical manifestations include sparkling, yellowish retinal crystals resembling chalk powder, choroidal atrophy and sclerosis, and yellow-white crystals in the superficial stroma and subepithelial layer of the peripheral cornea. Monoclonal gammopathy and multiple myeloma lymphoproliferative disorders may also cause crystalline keratopathy.16 Corneal accumulations may also form in the epithelium or stroma. Diagnosis is made by conjunctival biopsy, or blood or bone marrow smear. Drug-induced crystalline keratopathy may also occur with the use of fluoroquinolone (ciprofloxacin) drops. The effects usually resolve spontaneously when the topical fluoroquinolone is discontinued.Ocular cystinosis is considered one of the corneal crystalline keratopathies. The term crystalline keratopathy describes a group of diseases in which crystal deposits form on the anterior surface of the corneal epithelium or stroma due to a variety of reasons such as infection, corneal dystrophies, or systemic causes. The etiology of crystalline keratopathy may be ocular, systemic, or medication-induced. Infectious causes include 17 It initially manifests as asymptomatic aminoaciduria. Proximal tubular dysfunction which develops between 6-12 months of age leads to the loss of amino acids, sodium, potassium, bicarbonate, magnesium, carnitine, calcium, phosphate, glucose, and low-to-middle molecular weight proteins in the urine, resulting in the development of Fanconi syndrome.18 Infants may present with lack of appetite, polyuria, polydipsia, severe dehydration and electrolyte imbalance, vomiting, constipation, and sometimes vitamin D-resistant rickets. Biochemical tests may reveal hypokalemia, metabolic acidosis, hypophosphatemia, hypocalcemia, low carnitine level, and hyponatremia. If left untreated, end-stage renal failure develops by the end of the first decade.19Infantile nephropathic cystinosis accounts for 95% of cystinosis cases and is the most severe form of the disease. The renal phenotype consists of Fanconi syndrome, in which progressive glomerular dysfunction and loss eventually lead to end-stage renal failure.20 This form is less severe than nephropathic cystinosis, with milder disease course and symptoms. Children with juvenile cystinosis do not exhibit significant retardation of growth or development.Five percent of cystinosis patients are diagnosed with the juvenile form. It appears in late childhood or early adolescence.21 Patients exhibit subclinical hypothyroidism characterized by normal T3-T4 levels and elevated TSH.In 50-70% of patients, fibrosis resulting from accumulation of cystine crystals in thyroid follicular cells causes primary hypothyroidism in the second decade of life.22 Around the age of 18, 50% of infantile cystinosis patients develop gradual reduction of insulin secretion and C-peptide production, resulting in glucose intolerance and diabetes.Endocrine and exocrine pancreatic insufficiency often occurs in patients with cystinosis after renal transplantation.23 Azoospermia may cause infertility. Female patients are usually asymptomatic.Primary hypogonadism occurs in 70% of male cystinosis patients.23 Cerebral cortical atrophy, hydrocephalus, demyelination, and vacuolar necrotic brain changes may develop at later ages.Central nervous system involvement may cause hypotonia, tremors, delayed speech, gross and fine motor impairment, idiopathic intracranial hypertension, neurocognitive dysfunction, behavioral disorders, and encephalopathy.The cysteamine used in the treatment of cystinosis acts by disrupting the disulfide bonds of the cystine molecule, allowing the resulting intermediary metabolites to be eliminated from the body without accumulating. Topical cysteamine therapy administered in ocular cystinosis aims to reduce the cystine crystals that have accumulated in the cornea and to prevent complications they may cause. It has been shown that treating patients for 6 months with topical 0.05% cysteamine drops administered 5 times daily resulted in reduction of corneal cystine crystals, improvement in visual acuity, and decreased photophobia and blepharospasm. However, there was no decrease in corneal crystals after topical cysteamine use in our patient.Cystinosis is a metabolic disease that causes the accumulation of cystine crystals throughout the body and most commonly affects the eyes and kidneys. Ophthalmologists play an important mediator role in diagnosing asymptomatic cystinosis patients by anterior segment examination, especially during routine eye examination. Patients diagnosed with cystinosis should be referred to pediatric nephrology for systemic involvement and possible complications. Early initiation of cysteamine therapy is beneficial for preventing the development of late-stage renal failure.


Dear Reader,Sports Medicine - Open over the past 12 months.As we approach the end of 2016, we would like to thank all those who have contributed to Sports Medicine - Open over the course of 2016. Their skill and dedication, along with their support for this journal, are critical to the journal’s success.Firstly, we must acknowledge how much we are indebted to all of the authors who have submitted articles to Sports Medicine - Open over the last 12 months:The quality of the articles published is also testament to the significant efforts of the peer reviewers, whose commitment ensures that the journal’s content is held to the highest possible standard. We would like to thank the following individuals who, in addition to members of the Honorary Editorial Board, acted as reviewers for articles published in Maicon AlbuquerqueBrazilMaurice ArnaudSwitzerlandPatricia AronsonUSAGuilherme ArtioliBrazilGreg AtkinsonUKTodd BackesUSASeung-Soo BaekRepublic of KoreaTyson BeachCanadaClint BellengerAustraliaStephen BirdAustraliaBarbara BowsherUSAMichel Brinkthe NetherlandsSiacia BroosBelgiumRoland BüchterGermanyDavid BuzasUSALilian Calderón-GarcidueñasUSAKarina Rabello CasaliBrazilCristina CasalsSpainRicardo CassilhasBrazilHelmi ChaabèneTunisiaHamdi ChtourouTunisiaFrancis ConidiUSAStephen CornishCanadaRobert DefraitesUSAKevin DeightonUKDominic DellwegGermanyDavid DochertyCanadaKenji DomaAustraliaSladjana Dronjak-ČučakovićSerbiaMeghan EdwardsUSAMatthew ElyUSADamian FarrowAustraliaPeta ForsythAustraliaPaul FreemanUKVictor FroelicherUSAThomas GeeUKBruno GrassiItalyTom HazellCanadaGenevieve HealyAustraliaMartin HoffmanUSASharmin HossainCanadaJohn HoughUKErin HowdenAustraliaChun-Jung HuangUSAJackie HudsonUSAFlorentino HuertasSpainCharlotte Huppertzthe NetherlandsStefanie HüttermannGermanyShigeki IzumiJapanMichael JaffeeUSASaša ŠegaJazbecSloveniaBruce JonesUSAKelly JonesNew ZealandMariana KaiselerUKKristof KippUSAThomas KjeldDenmarkBeat KnechtleSwitzerlandKarsten KoehlerUSASylvain LabordeGermanySteen LarsenDenmarkDavid LavalleeUKTrent LawtonNew ZealandJohn LeddyUSASeth LenetskyNew ZealandDuncan MacfarlaneHong KongKarin MagnussenDenmarkYorgi MavrosAustraliaScott McIntoshUSATerry McMorrisUKRomain MeeusenBelgiumRégis MestrinerBrazilGeoffrey MinettAustraliaLluís MontSpainColleen MuñozUSAMarius MyrstadNorwayTamas NagyHungaryFabio NakamuraBrazilDavid NiemanUSAOnni NiemeläFinlandVahur ÖöpikEstoniaSushil PantNepalCamille PerchouxFranceNoel PerkinsUSANeal PollockUSADenis Prud'hommeCanadaJoseph PuritaUSAClaudio Rafols Del CantoChileAnette RanhoffNorwayUlrike RimmeleSwitzerlandRodrigo RodriguesBrazilKelly RussellCanadaBryan SaundersBrazilB. Volker ScheerUSAMarc Scheltingathe NetherlandsAlison Schinkel-IvyCanadaStephen SimonsUSADean SimontonUSAJonathan SingerCanadaNeil SmartAustraliaJostein Steene-JohannessenNorwayUrs StöckerSwitzerlandMogammad Sharhidd TaliepSouth AfricaMarcus TaylorUSAMatthew TuckerUSALuc van Loonthe NetherlandsWill VickeryUKHenry WillifordUSAPeter WilmshurstUKCarl WoodsAustraliaLéder XavierBrazilWanghong XuChinaJulia ZakrzewskiUKFinally, we would like to offer our sincere thanks to the members of the journal’s Honorary Editorial Board, who have acted as peer reviewers and authors, and have provided guidance on journal content, policy and processes:Duarte AraújoPortugalPeter ArnettUSASam BlackerUKFadi CharcharAustraliaJia Yi ChowSingaporeMatias Costa-PazArgentinaKeith DavidsUKJoel GagnierUSARuben GoebelQatarJohn HawleyAustraliaMartin HägglundSwedenPatria HumeNew ZealandMikel IzquierdoSpainSteve KamperAustraliaJustin KeoghAustraliaTom LoneyUnited Arab EmiratesTim MeyerGermanyCarles PedretSpainLuis PratoUSAMichael SchwenkGermanyAlison SeptemberSouth AfricaSanjay SharmaUKRoy ShephardCanadaRoberto SimãoBrazilKeith TolfreyUKWillem van Mechelenthe NetherlandsMounir ZokUSAWe hope that you have found the articles published throughout 2016 to be both interesting and informative. We have appreciated the high quality of content contributed to the journal this year and we look forward to keeping you up to date with sport and exercise science research in the year to come.With best wishes,Steve McMillan & Roger OlneyEditors


Tuberculosis (TB) is a global health burden, and a major cause of mortality and morbidity in West Africa. Here, we select keyconserved pathogen epitopes of proven immunogenicity to form a potential TB epitope ensemble vaccine. We compared two vaccineformulations: one comprising class I epitopes from the 13 most prevalent class I epitope-bearing antigens and class II epitopesderiving from the 20 most prevalent class II epitope-bearing antigens and another consisting of epitopes derived solely from 5 antigensidentified as the most immunogenic by VaxiJen. In the prevalence analysis, 279 class I and 561 class II epitopes were collected and asubset selected using our published methodology, yielding 32 conserved epitopes. Combining 9 conserved epitopes gave a putativevaccine with predicted population coverage (PPC) over 95%. This consists of ISSGVFLLK, AVAGAAILV, WYYQSGLSI, YQSGLSIVM,MPVGGQSSF, QSSFYSDWY, WDINTPAFEWYYQSGLSIVM, DAPLITNPGGLLEQAVAVEE and NQAVTPAARALPLTSLTSAA. 5immunogenic antigens VaxiJen-identified yielded 187 epitopes, which we again analyzed using published protocol. This identified 11conserved epitopes. From this set the highest PPC value (<85%) was obtained by combining: GQQYQAMSAQAAAFH,DDIKATYDKGILTVSVAVSE and AVAGAAILV. We conclude that prioritizing epitope selection using predicted immunogenicityalone is likely to be unduly restrictive and is currently not an optimal or advisable strategy in the design of epitope ensemble vaccines. Mycobacterium Tuberculosis (TB) is a global health burden withan estimated 2015 death toll of 1.8 million, with 10.5 million newTB cases .The immune response against TB predominantly involves T cellmediated immunity, specifically CD4+ and CD8+ T cells, whichbind to complexes of class I or class II MHC bound to specificepitopes. Once activated, both CD4+ and CD8+ cells secretecytokines inducing an immune response. CD8+ T cells alsomediate cytotoxicity and the lysis of infected cells.The Bacillus Calmette-Guérin (BCG) anti-TB vaccine has beenused for disease prophylaxis and control of TB progression forover 80 years. BCG is an attenuated form of mycobacterium bovisand is currently the only anti-TB vaccine in use worldwide.However, BCG has a well-attested variable efficacy (0-80%) inadult populations presenting with pulmonary TB , limitinSeveral live vaccines are currently in development against TBincluding recombinant BCG VPM1002 and MTBVAC . A keyiHere, we seek to identify pathogen epitopes able of form anepitope ensemble vaccine against TB. The complete genomesequence of Mycobacterium Tuberculosis H37Rv strain,comprising over 4 million base pairs and approximately 4,000genes  andinflWe applied an epitope ensemble vaccine design protocol alreadydescribed ,9 to TB.TB Antigens were processed using VzxiJen (URL:http://www.ddgpharmfac.net/vaxijen/Vaxijen.html), whichpredicts immunogenicity scores, allowing classification ofantigens as either an antigen capable of inducing protectiveimmunity or as a non-antigen ,11. BactTwo sets of epitope-bearing antigens were collected. The firstdataset comprised proteins with greater than 10 epitopes. Therewere 13 Class I epitope-bearing antigens (Ag85B (59 class Iepitopes), Putative ATP-dependent 6-phosphofructokinaseisozyme 2 (28 epitopes), Alanine and proline-rich secretedprotein Apa (26), Probable membrane protein Rv1733c (26),ESAT-6-like protein EsxB (23), ESAT-6-like protein EsxH (19),Low molecular weight T-cell antigen (17), 6 kDa early secretoryantigenic target (16), Lipoprotein LpqH (14), PPE68 (14), Ag85A(13), Alpha-crystallin (13), MPT64 (11)) and 20 class II-bearingantigens (Ag85B (55 class II epitopes), 6 kDa early secretoryantigenic target (55 epitopes), ESAT-6-like protein EsxB (45), 60kDa chaperonin 2 (42), Uncharacterized PPE family proteinPPE19 (36), Ag85A (34), Putative ATP-dependent 6-phosphofructokinase isozyme 2 (32), Alanine and proline-richsecreted protein Apa (30), Phosphate-binding protein PstS 1 (29), Uncharacterized PPE family immunogenic protein PPE68 (27),MPT64 (24), Alpha-crystallin (22), Lipoprotein LpqH (21),Probable membrane protein Rv1733c (21), MPT70 ( 19), 10 kDachaperonin (18), MPT83 (15), Putative phthioceroldimycocerosate transporter LppX (12), ESAT-6-like protein EsxH(12), MPT63 (12).PVS identified conserved epitopes. From the 13 class I epitopebearingantigens, only 9 had conserved epitopes. Overall, 22conserved epitopes were found. The greatest number of epitopeswas collected from diacylglycerol acyltransferase Ag85B (11),followed by lipoprotein LpqH (3), diacylglycerol acyltransferaseAg85A (2), alanine and proline-rich secreted protein Apa (2),PPE68 (1), ATP-PFK 2 (1) and alpha-crystallin (1). From the 20class I epitope-bearing antigens, only 5 had a total of 11conserved epitopes: ATP-PFK 2 (1), alanine and proline-richsecreted protein Apa (1), lipoprotein LpqH (1), diacylglycerolacyltransferase Ag85B (3), PPE19 (5).The second dataset comprised antigens with 3 or more epitopesfor analysis using VAXIJEN. For class I, 22 antigens wereidentified. Antigens with the top 5 immunogenicity scores wereidentified: Lipoprotein LpqH ,Isoniazid-induced protein , ESAT-6-like protein EsxB, Alpha-crystallin , PPE68 . Forclass II, 61 antigens were identified. Antigens with the top 5immunogenicity scores were identified: PE_PGRS14 , Lipoprotein LpqH , ESAT-6-like proteinEsxB , Alpha-crystallin ; PstS 1 .PVS identified conserved epitopes: for class I only 4 antigens hadconserved epitopes, returning a total of 6 epitopes, and for class IIonly 3 of the 5 antigens had conserved epitopes, returning a totalof 5 epitopes.For both VaxiJen and non-VaxiJen analysis, combinations ofepitopes were assessed for population coverage. For the high prevalenceset, non-VaxiJen analysis, the highest PPC valuefound was 98.75  and 95.60 (West Africa). For the VaxiJenanalysis, the highest PPC value found was 83.79  and68.62 (West Africa). See Compared to the unrestricted analysis, results from VaxiJenranking were disappointing. Due to the small number ofconserved epitopes identified, limited combinations could beundertaken. The conserved epitopes from VAXIJEN have similarHLA binding profiles. For class I, A*02:06, A*02:01 and A*02:03were the most common restrictions seen. The limited variation inbinding profiles contributed to the low PPC values seen. Thehighest combined PPC values for both subsets contained the classI conserved epitope AVAGAAILV, derived from lipoproteinLpqH, consistent with the epitope being highly immunogenicand promiscuous.In this vaccine design exercise, we sought to limit selection toepitopes from major highly immunogenic antigens. Suchconstrained selection should concentrate the immune responseon antigens more likely repeatedly to evoke strong responses invivo. As an alternative to using all experimentally verifiedepitopes available, we used VaxiJen - the first and, ostensibly, theonly on-line server for whole protein immunogenicity prediction- to rank antigens in terms of potential efficacy. However,prioritizing antigens using VaxiJen did not prove effective. Thisidentifies a shortcoming of this method, due in no small part tothe dearth of adequate training data. This is something we willseek to rectify in due course. Prevalence analysis, however,allowed for the total pool of available epitopes to be tractable yet large enough to generate an optimal hypothetical vaccinecandidate. Epitope combinations derived through epitopeprevalenceanalysis had sufficiently high population coverage togenerate effective potential universal epitope ensemble vaccines.However, due to the low immunogenicity traditionallyassociated with peptide vaccines, such candidates are likely torequire the addition of adjuvants. Overall, we have shown howcomputational approaches can identify a potential ensemblevaccine against a major bacterial pathogen by selecting epitopesof known, validated immunogenicity with broad populationcoverage. These will now need experimental validation todetermine the efficacy and safety of the vaccine in live subjects.


This issue closes out Volume 3 of the JACMP. I am pleased to note that the journal continues to grow in size and outreach. Volume 2 was 60% larger (both in the number of manuscripts and the number of pages published) than Volume 1 and Volume 3 is 60% larger than Volume 2. In three years the journal has more than doubled in size.The monthly usage statistics for the journal have also shown a steady increase, both in requests and in the unique hosts accessing the journal, during these three years.Of particular interest has been the number of different countries from which manuscripts have been submitted. In addition to the United States and Canada, papers have come from fourteen others countries. Although the majority of the manuscripts come from the United States the percentage of submissions from other countries has increased in the last few months. Because an electronic journal is accessible worldwide it has always been editorial policy to serve the international medical physics community, and it is gratifying to see this taking place.The success of the journal and its growth would not have been possible without the continued support of the American College of Medical Physics, who along with the journal's sponsors and advertisers have continued to underwrite the cost of the journal. This means that in 2003 Volume 4 will again be subscription free and available to all medical physicists.The completion of Volume 3 also marks the end of my tenure as Editor‐in‐Chief and that of Georgeanne Moore as Manuscript Manager. Much of the success of the journal has been due to her hard work and handling of the daily mechanics of running the journal. The journal is very fortunate that Dr. Edwin McCullough will be taking over as Editor‐in‐Chief and Sarah Evers as the Manuscript Manager. We wish them well and look forward to continued growth and success of the journal in their capable hands.Peter R. Almond, Ph.D.Editor‐in‐Chief


Journal of Comorbidity and the Society for Academic Primary Care (SAPC) are pleased to announce an exciting new partnership aimed at strengthening collaborations and enhancing opportunities among primary care professionals with an interest in comorbidity and multimorbidity. The Journal of Comorbidity and SAPC share a mutual goal to improve the management and care of patients by making clinical and research information and perspectives available to a global network of healthcare professionals. This new partnership will be an invaluable contribution to expanding the research platform for discussions and the scholarly exchange of knowledge, ideas, and research on comorbidity and multimorbidity.The SAPC provides a clear voice and a strong presence for academic primary care in the complex and ever-changing primary care environment. The Society offers a point of reference and contact for those seeking academic solutions to the problems they face in the provision and study of primary care, and the advancement of academic primary care.Academic general practice was present from the inception of the UK National Health Service (NHS), and the Society has been at the heart of that community for over four decades. The first independent academic department was established at the University of Edinburgh in 1956. Other departments emerged, and in 1972, held their first scientific meeting. In 1974, a new society  was formed. A growing recognition of the role and contribution of the wider academic primary care community led to the group changing its name to the Society for Academic Primary Care in 2000 [To facilitate this endeavour, the SAPC holds annual and regional conferences that bring together hundreds of researchers and educators from the primary care community to showcase their latest work on all aspects and issues relating to primary care. The annual conference not only attracts delegates from the UK but also includes professionals from across the globe who share their experiences and research findings, attend workshops and special interest sessions, and network with like-minded primary care professionals. More recently, the annual SAPC conference has become an important avenue for discussing and disseminating research on multimorbidity.Journal of Comorbidity comes at a historic time, as multimorbidity is now the norm, not the exception. Understandably, interest in and awareness of multimorbidity are growing, but much more knowledge, research, and guidance on the care, treatment, and understanding of multimorbidity is desperately needed. SAPC has had a long-standing interest in, and is committed to, building affiliations between the academic community and publishers with primary care interests. This work contributes to the three pillars of SAPC activity to:The partnership with the Raise the profile of academic primary care and SAPC to a wider audienceSupport workforce career development, including increased access to various publishing opportunitiesEnhance impact through collaboration.Journal of Comorbidity’s mission, which is to provide a high-quality resource to fulfil the following objectives:These pillars closely align with the Highlight comorbidity/multimorbidity as a fundamental component of overall patient careEnhance the understanding of comorbidity/multimorbidityIdentify and fill gaps in the evidence on comorbidity/multimorbidityProvide comprehensive guidance on preventing and managing comorbidity/multimorbidityPromote the collaboration of multidisciplinary healthcare providersFacilitate daily decisions in clinical practice.Journal of Comorbidity. It is just the beginning of SAPC’s renewed efforts to increase publishing collaborations and to provide its members with a broader platform of engagement with a key partner who shares our goal to provide useful and accessible health information. We look forward to working with the journal’s Editors in widening the reach, impact, and engagement with key researchers in the field of multimorbidity.SAPC is delighted to be working in partnership with the Journal of Comorbidity is an international, open-access, peer-reviewed journal that aims to optimize the management of patients with comorbidity and multimorbidity. The journal features original clinical and experimental research articles, guidelines, policies, editorials, commentaries, protocols, and critical review articles, as well as proceedings of congresses. The Editorial Board also welcomes ideas and suggestions for special issues dedicated to unique themes.The Journal of Comorbidity and SAPC is an important move in encouraging SAPC members to consider the Journal of Comorbidity as the premier journal for publishing their work related to comorbidity and multimorbidity. As a benefit of this partnership, SAPC members will receive a discounted rate to publish in the journal.The affiliation between the The Editorial Board is very excited to partner with SAPC and look forward to working with them over the coming years, and to the active participation of its members. We are confident that the journal, in partnership with SAPC, will continue to grow and ultimately increase its impact in this important research field.