To quantify long-term changes in stromal collagen ultrastructure following penetrating keratoplasty (PK), and evaluate their possible implications for corneal biomechanics.A pair of 16 mm post-mortem corneo-scleral buttons was obtained from a patient receiving bilateral penetrating keratoplasty 12 (left)/28 (right) years previously. Small-angle x-ray scattering quantified collagen fibril spacing, diameter and spatial order at 0.5 mm or 0.25 mm intervals along linear scans across the graft margin. Corresponding control data was collected from two corneo-scleral buttons with no history of refractive surgery. Wide-angle x-ray scattering quantified collagen fibril orientation at 0.25 mm ×0.25 mm  intervals across both PK specimens. Quantification of orientation changes in the graft margin were verified by equivalent analysis of data from a 13 year post-operative right PK specimen obtained from a second patient in a previous study, and comparison made with new and published data from normal corneas.Marked changes to normal fibril alignment, in favour of tangentially oriented collagen, were observed around the entire graft margin in all PK specimens. The total number of meridional fibrils in the wound margin was observed to decrease by up to 40%, with the number of tangentially oriented fibrils increasing by up to 46%. As a result, in some locations the number of fibrils aligned parallel to the wound outnumbered those spanning it by up to five times. Localised increases in fibril spacing and diameter, with an accompanying reduction in matrix order, were also evident.Abnormal collagen fibril size and spatial order within the PK graft margin are indicative of incomplete stromal wound remodelling and the long term persistence of fibrotic scar tissue. Lasting changes in collagen fibril orientation in and around PK wounds may alter corneal biomechanics and compromise the integrity of the graft-host interface in the long term. The biomechanical properties of the cornea are influenced significantly by the organization of collagen fibrils that form the bulk of the corneal stroma Here we have used x-ray scattering to quantify stromal collagen ultrastructure in a pair of post-mortem human eyes from a patient who underwent bilateral PK for keratoconus 12 (left eye) and 28 (right eye) years previously, and a 8.5 mm post-operative corneal button from the right eye of a second patient who required re-graft 13 years after originally undergoing PK. The availability of tissue of this kind is extremely limited. This study therefore afforded us a rare opportunity to investigate the long-term effects of penetrating injury on corneal ultrastructure by carrying out the first quantitative study of collagen organisation across whole corneal specimens featuring PK wounds.The research presented in this manuscript was approved by the Human Science Ethical Committee  and the South East Wales Research Ethics Committee . The institutional review board approved the use of all corneas described in this study; a waiver of consent was given for the donor corneas. All tissue used in this study was obtained in accordance with the tenets of the Declaration of Helsinki, and local ethical rules were adhered to throughout. All experimental procedures were performed in accordance with the Declaration of Helsinki.Left eye history: Underwent 8.2 mm/8.0 mm PK for keratoconus 12 years previously. The apex was thinned and located inferior/temporal. However, thinning and astigmatism could not be accurately quantified due to irregularity of the corneal surface. 5 months after PK there was vascularization into the donor cornea as far as the sutures. The sutures  were removed after 12 months, and at 14 months there were no reported complications. Right eye history: Underwent PK for keratoconus 28 years previously. No further information was available.A pair of post-mortem eyes with history of bilateral PK for keratoconus was obtained from The Danish Cornea Bank, Aarhus University Hospital . The age of the PK recipient at death was 79 yrs. The donors' ages were not available. 16 mm diameter corneo-scleral buttons were excised and the orientation within the eye marked with a scleral suture. The specimens were then immediately wrapped in polyvinylidene chloride film to prevent dehydration, frozen and stored at −80°C until the time of x-ray experiments. Inspection of the PK tissue using light microscopy prior to the x-ray experiments disclosed that both specimens displayed prominent regions of opacity in the peripheral cornea, located on the temporal side in the left eye and inferior/superior in the right, and that the graft margins were still visibly demarcated . AdditioA 8.5 mm diameter post-operative corneal button from a 51 year old patient who required re-graft following suspected recurrence of keratoconus 13 years after initial 7.5 mm/7.25 mm PK surgery. A detailed structural examination of this specimen was published by our lab previously and concluded that corneal ectasia at 13 years may have recurred due to a mechanical failure of the graft rather than keratoconus recurrence Two post-mortem corneo-scleral buttons from donors aged 71 yrs (Control 1) and 47 yrs (Control 2), with no history of keratoconus or refractive surgery, were obtained from Bristol Eye Hospital . The specimens were wrapped in polyvinylidene chloride film, frozen and stored at −80°C until the time of x-ray experiments. Further control results were obtained via re-analysis of raw x-ray data from a normal cornea (Control 3) examined in detail in a previous publication by our lab Small-angle x-ray scattering Following SAXS experiments, the PK specimens were immediately placed in 4% paraformaldehyde and stored at 4°C for subsequent characterization using wide-angle x-ray scattering (WAXS). Our previous work has established that this mild fixation method does not affect corneal/scleral collagen parameters as measured by WAXS Analysis of corneal/scleral WAXS patterns provides a quantitative measure of bulk collagen fibril orientation, as an average value within the stromal volume sampled by the x-ray beam Firstly, the circumferential collagen which characterises the normal human limbus and perilimbal sclera had extended into the peripheral cornea on the temporal side, corresponding in location with the region of prominent corneal opacity  - arrow.inset). These results are shown in In order to quantify further the change in normal alignment of collagen across the graft margin, we compared the relative proportions of collagen oriented transversely and parallel to the wound edge. This was done by calculating from the WAXS patterns Although PK is a highly successful long-term treatment option in cases of severe corneal scarring and advanced pathology, late-onset complications can present in a minority of patients. In the case of PK for advanced keratoconus, the indication presented by the patients studied herein, graft failure occurs in around 6% of cases at a mean time of 13 years post-op Significant changes in collagen alignment around the wound margin could impact on the mechanical stability of the graft-host interface. Specifically, at the examined location in the left PK graft margin of Patient 1, our measurements indicated that only around 10% of the total collagen was aligned transversely to the wound edge. Such a relative lack of meridional fibrils spanning the graft margin might be reasonably expected to encourage a wound under tension to reopen. In contrast, over 50% of the collagen at this point was aligned parallel to the wound edge. Notably, a predominantly circumferential arrangement of fibrils is predicted, from biomechanical considerations, to maximise out-of-plane corneal deformation under intraocular pressure Although neither eye of Patient 1 examined herein suffered a reported failure, the wound margin of the patient's right eye did display a prominent visible separation on the temporal side, which was confirmed by the collagen orientation pattern in this region. It may be significant that this graft was performed earlier and hence had been in the eye some 16 years longer than the fellow eye, which showed no such changes. Moreover, we also noted a distortion of the collagen in the tissue between the separated wound edges, in which fibrils had aligned radially, i.e. along the direction of the apparent outward migration of the wound edge and perpendicular to the edge itself. The altered collagen alignment here may not have resulted in a significant local mechanical effect because the degree of anisotropy was generally low (indicated by the orange color coding of the vector plots). However, interestingly a similar pattern of radial collagen alignment abutting the wound edge was previously noted by the same technique in the specimen from Patient 2, in which an initially successful PK graft had seemingly mechanically failed after 13 years The long-term persistence of circumferential collagen in the graft margin we report here is compatible with the general view of stromal collagen after penetrating corneal injury having a limited capacity to remodel following initial wound repair, and this is further supported by our observation of altered collagen spatial organisation in the graft margin of Patient 1. Specifically, we noted localised increases in collagen spacing and diameter, and a reduced level of spatial order, observations which are all consistent with the presence of fibrotic scar tissue deposited early in wound healing A final observation of the current study of potential importance to corneal biomechanics is that, with reference to the dominant fibril directions of the donor and host collagen, the right PK donor button appeared to have been grafted obliquely. It has been suggested that the superior-inferior and nasal-temporal preferred collagen orientation in the central human cornea may reflect a mechanical adaptation of the tissue to forces imposed by the extraocular muscles The peripheral (keratoconic) tissue of both eyes of Patient 1 displayed prominent peripheral corneal opacity, located temporally in the left eye and inferior/superior in the right. The WAXS data from this these regions revealed abnormal levels of highly aligned tangential collagen, resembling that normally restricted to the limbus and perilimbal sclera In summary, this paper documents the first quantitative, long-term follow-up of collagen organisation across whole transplanted corneas. In principle, the changes noted in and around the graft margin could affect corneal biomechanical behaviour and graft stability. However further research, possibly focussing on the application of structural data in numerical simulation of corneal biomechanics, may help to establish any definitive link that may exist between the ultrastructural changes presented herein and the instances of PK graft dehiscence and mechanical failure documented in the clinical literature.


Only by gazing, thus, can the Alpine climber find values and often he catches too a glimpse of beauty.”[ beauty.”The Indian Journal of Psychiatry (IJP) in its multiple avatars has completed 60 years of existence. For an Institution like the Indian Psychiatric Society with IJP at its core, this is relatively a very small step. It has many miles to go and many more milestones to surpass. However,The dawn of Indian independence also saw the birth of the Indian Psychiatric Society (IPS) and may be regarded as the beginning of the modern psychiatric movement in the country. In the sFrom the very beginning the IJP followed patterns used in reputed international journals containing sections such as editorial, original articles, reviews news and abstracts. De 1949-1951) in his first editorial dealt with the opportunities in psychiatric research in India, opening his remarks with, “Research is the life of Science. A Science is living only as long as research is carried out in it.”[1 in his The first year had an extensive review article titled “Mental Health Services in India” by N. N. De. Most of the papers published then dealt with theoretical issues with a strong inclination towards psychoanalysis and other disciplines of psychology. The referencing style conformed to that used in Quarterly Cumulative Index Medicus.De resigned on grounds of ill health and L. P. Varma. 1951-1954) took over as Editor. Varma edited three volumes of the journal, bringing out a total of nine issues, some of which were combined . Among the outstanding works done during this period were excellent reports on the phenomenology of general paresis of insane; interest851-1954 tAfter Varma, M. V. Govindaswamy was elected its editor 1954-1958), but the journal ceased publication[-1958, buThe revival of the Indian Journal of Psychiatry in its present name was decided at the annual meeting of the IPS in 1958 at Poona under the leadership of I.K. Mujawar. A council under M. V. Govindaswamy was to undertake necessary actions to its resumption. However Dr. M. V. Govindaswamy resigned as elected editor and Col P.N. Bardhan (1958-1960), a pathologist, became the editor and the first issue of the “Indian Journal of Psychiatry” was printed in October 1958 .In the first editorial of the IJP, Col. Bardhan wrote, “The first series was called Indian Journal of Neurology and Psychiatry and there is significance in the change of the name of the journal. Both neurology and psychiatry are major specialties in their own right and a separate journal could easily be devoted to each. The first series was discontinued owing to unavoidable reasons, ill health none the least” and concluded with “the field of psychiatry in this country is very vast and ever expanding. Therefore, to justify its existence, this journal must represent the experience and progress in every branch of psychiatry”.The journal followed the same format as the earlier journals, but the referencing pattern changed to Harvard type and a supplement continued to accompany the first issue of the journal which included details about the meetings and members of various committees of the society.The first 25 years of the IJP mirrored the growth of psychiatry and there were many factors which influenced improved research and publication environment in the country. A remarkThe IJP has since been in the forefront of publishing research by Indians and helped stimulating psychiatric thinking and research in the country. It has, Outstanding work was done during this phase in the field of epidemiology, phenomenology, and therapeutics. The focus of interest seems to have shifted from individual to mass problems and from man to society in general. There were excellent surveys of mental morbidity in general population by researchers from all over the country.–16 Large22This remarkable interest in sociology of psychiatric disorders and in psychology of social phenomena may be attributed to the general enthusiasm, among the psychiatrists for action. This was coupled with either of the following two factors, namely: lack of resources to conduct laboratory research of any kind as well as the effect of changing foci of interest and attention from couch to the community in Western psychiatric world.The mistaken impression that epidemiological work is easy to undertake and quick to bear results may also have played some role in shifting the interest towards mass studies. The changed political atmosphere of the country and socialistic policies of Indian Government may also be responsible, though on a somewhat unconscious level, for this shift of attention. In addit3137A few interesting papers on the relationship of crime with mental disorder–43 have Something that becomes quite obvious while reviewing the outstanding works, especially the ones in the second phase, is the preoccupation and attempts at, what one may call, “Indianization” of Psychiatry, Phenomenological studies with a tendency to explain the form and content of symptomatology in relation to Indian culture, for examThese epidemiological, phenomenological and therapeutic studies, which attracted researchers in the 60s and 70s continued well into the 80s. In additThis initial phase saw a total of four editors viz. Col P. N. Bardhan, (1958 -1960), M. R. Vachha, (1961-1967), Venkoba Rao (1968-1976) and B. B. Sethi (1977-1984), who was, supported by Rudraprakash as assistant editor. Col Bardhan was a pathologist, Vachha a neurologist and Venkoba Rao the first psychiatrist to be editor of the IJP. The IJP crossed significant milestones during their tenures.Prashanti written in Hindi characteristics.[In 1961, the IPS got a new emblem with a brain and a staff of Aesculapius carved on a potter wheel with eristics. Under theristics. Under theristics. The sameeristics.In this period there were a total of eight editors with three of them lasting for less than a term of two years. This churning was more in the last five years of this period. The teams of earlier editors and assistant editors were S. M. Channabasavanna (1985-1988) with S. Chaturvedi, A K Agarwal , the front page was again changed and the current format was used for the first time. Under J. K. Trivedi, the IJP was available in electronic format and linked to the newly formed website of the IPS. Under N. G. Desai, the IJP underwent major changes in the format which is still being followed. The referencing style of the journal was reviewed again and for the first time the Vancouver System was adopted.Schizophrenia, major depression, substance abuse, organic disorders, psychiatric aspects of medical disorders and anxiety disorders respectively formed a major part of the publication in the IJP during this period.58 The arStudies on schizophrenia concentrated on phenomenology,–60 cours6366Studies on Depression and Bipolar Disorder too have concentrated on phenomenology, course,–72 disab74In comparison to the earlier quarter century, the number of articles on the elderly has incrStudies on Addiction medicine, especially alcohol–81 as we8865While drug trials predominated in the first 25 years, these dwindled during the next phase. Most drug trials were replications of those from the West, but helped evaluate efficacy and adverse effects in the Indian population.–99 Howev100103111Psychiatric education has attrwww.journalonweb.com) is now in place. This has reduced clerical work as well as submission-decision time and thanks to this system we now have about more than 200 articles in various stages of review or publication.[The year 2007 saw three major transformations, all interrelated, in the journal. M/s Medknow Publications was appointed publisher, printer and distributor of the IJP who gave the journal access to the other two changes. An online manuscript submission and electronic peer review system (One of the important advantages of the system is that the search for reviewers who are key researchers in the field can be identified from the references quoted in the article as well as a PubMed search using key words. Getting articles reviewed by such significant authorities has been an enriching experience which has greatly improved the worthiness of the publication. It is imwww.indianjpsychiatry.org) which contains full text articles has helped avoid the time lag between decision and printing and the articles have been available for viewing well before it arrives in the hands of subscribers.[The second important transformation is the posting of the journal well before printing on the website  at Kolkata, which was well appreciated. The release of the IPS Golden Jubilee Commemorative volume which contained 50 presidential addresses and 50 editorials was another important feature. This volume was greatly cherished by members as it gawww.indianjpsychiatry.org. The DVD, in addition, contained the Clinical practice guidelines of the IPS, as well as the updated membership directory of the IPS. The IJP now has its articles available “full text online, fully archived”.[In 2009, a mammoth task of digitalizing all past issues of the Indian Journal of Neurology and Psychiatry (1949-1954), the Indian Journal of Psychiatry (1958-2008) was successfully undertaken and a DVD was distributed to all members during ANCIPS in January 2009 as well as archived in the IJP website rchived”.Thanks to these transformations, the journal is now indexed with various indexing agencies such as SCOPUS, DOAJ, Index Copernicus, Health and Wellness Research Center, Health Reference Center Academic, InfoTrac One File, Expanded Academic ASAP, Genamics JournalSeek, Ulrich’s International Periodical Directory, EBSCO Publishing’s Electronic Databases and Google Scholar. The receThe easy and universal access of all articles of the IJP from 1949 till date is a mine of information and this volume - “Annotations of Indian Psychiatry” is a natural extension. This volume is a compilation of articles on important research and publications by Indians, provides an insight to the achievements as well as the future of Indian Psychiatry.The IJP still has a long way to go as we await the impact factor rating from Thomson Science Citation Index. We do hope this will come sooner than later and with a good weightage.Psychiatric journals are both the means and repository of dissemination of professional learning. Journals


Diabetic Retinopathy for the comprehensive ophthalmologist. Walker J.http://drcobook.com. Please note that the book is now three years old and that new information has become available, in particular about intravitreal injections. However, the book also covers timeless topics such as informed consent and diabetes control.This book will be available in electronic format on the new Community Eye Health Update CD, due out with the December 2011 edition of this journal. Look out for your free copy! The book is also available for purchase  or free download (nine PDF files of 14-59MB each) from Diabetes grading scheme: International Clinical Diabetic Retinopathy And Diabetic Macular Edema Disease Severity Scales. International Council of Ophthalmology, October 2002.http://archive.icoph.org/standards/gdrp.htmlPatient information about diabetic retinopathy  www.retinalscreening.nhs.uk/userFiles/File/diabeticRetinopathyFacts.pdfPatient information about a DR screening programme: examples in several languages  www.dhsspsni.gov.uk/public_health_diabetic_retinopathyDetailed patient information about screening for DR  www.retinalscreening.nhs.uk/userFiles/File/EyeScreeningForDiabetes.pdfPatient information about laser treatment www.retinalscreening.nhs.uk/userFiles/File/PreparingForLaserTreatmentDR.pdf


This article explores reported practices of, and motivations for, diversion of methadone and buprenorphine, in a group of imprisoned individuals in OMT.28 in-depths interviews were conducted among 12 OMT-enrolled, imprisoned individuals, most of whom were remand prisoners. All had experienced tight control regimes prior to imprisonment due to varying degrees of 'non-compliance’ and illicit drug use during treatment. Their acquired norm of sharing with others in a drug using community was maintained when entering OMT. Giving one’s prescription opioids to an individual in withdrawal was indeed seen as an act of helping, something that takes on particular significance for couples in which only one partner is included in OMT and the other is using illicit heroin. Individuals enrolled in OMT might thus be trapped between practicing norms of helping and sharing and adhering to treatment regulations. ’Diversion’, as this term is conventionally used, is not typically understood as practices of giving and helping, but may nevertheless be perceived as such by those who undertake them.As we see it, the need to sustain oneself as a decent person in one’s own eyes and those of others through practices such as sharing and helping should be recognized. Treatment providers should consider including couples in which both individuals are motivated for starting OMT. Or we don't get any. You don’t always have some.While Erik accepted that he could not always obtain heroin, it was impossible for Mona to do so. She strongly feared withdrawal and said that she needed a steady income to ensure that she could always buy the heroin she needed. When she became involved in a relationship with a heroin-dependent man, higher income was needed. Even if he contributed, Mona did not have the security that she needed and explained that her choice was then between 3 “hells”: selling drugs, committing property crime or selling sex. She chose the latter because it gave her more control. Her male partner was included in OMT due to his worsening health status. He received a daily, supervised dose of methadone. Hence, he could not share his methadone when she was in withdrawal. They lived together, he in OMT and she on heroin. She continued to sell sex and regularly experienced violence and humiliation. This was a painful situation for both.In contrast to Mona and her partner, Simon was among those who found a situation in which only one partner was enrolled in OMT to be an impossible one. Before entering treatment, he had had a partner who was addicted to heroin and was clear about his opinion that a relationship in which one partner is in treatment and one dependent on illegal drugs involves unsolvable and unbearable moral dilemmas:Then you have to come up with ways [to give away your medication] to help your girlfriend if she is having a bad day. You can't just leave your girlfriend in withdrawal if you can help. So you're off to hell of a bad start if you can't both get help [treatment], together.The study participants had all committed serious crimes and reported criminal activity during OMT. They were all imprisoned at the time of the interviews, which may have influenced their decisions to share particular experiences and events and perhaps also their understandings of and retrospective reflections on these events and their consequences . All par“It is considered unethical to leave a person stranded when he or she is dopesick , unless one is openly feuding with that person”. We suggest that such communities are also communities of 'knowers’ – namely, people who possess corporeal knowledge of withdrawal and are thus, quite literally, able to understand the corporeal condition of another human being in that state. Importantly, the act of giving to a friend in withdrawal with known tolerance for opioids may have a lower harm potential than that of selling to unknown and potentially opioid-naive persons [The moral economy of sharing in drug cultures is well documented by the ethnographic work of Bourgois ,20 and t persons .When a heroin-dependent individual is included in OMT, he or she might struggle to navigate norms of different social systems: the treatment system with its external control measures and possibilities for 'sanctions’ for what is perceived as 'diversion’ and drug using communities with their values of civil and informal execution of what is perceived as 'sharing and giving’. The logics upon which these systems are based – namely, 'harm’ and 'helping’, respectively – might work against each other. OMT enrollment can hence place someone in a position in which he or she violates his or her norms of helping and sharing in order to follow treatment regulations. Diversion’, as this term is conventionally interpreted by clinicians, is not typically understood as practices of giving and helping. These practices may nevertheless be perceived as such by those who undertake them. For those who participated in this study, it was not as though norms for interpersonal relations maintained while using illegal drugs could be nullified when entering treatment. As we see it, the need to sustain oneself as a decent person in one’s own eyes and those of others should be recognized. Thus, the 'positive’ interpersonal skills and practices of OMT patients could perhaps be further explored as possible resources throughout the clinical encounter and rehabilitation process. Further, clinicians should encourage and support strategies developed by individuals in OMT to avoid diversion of their opioid prescriptions, such as changing from one dispensing pharmacy to another in an effort to avoid potential buyers in certain areas. Finally, treatment providers should focus on the patients’ social lives and indeed consider including couples if both individuals are motivated for starting OMT.OMT: Opioid maintenance treatment.The authors declare that they have no competing interests.TC was project manager. TC, IH and ALM conceived the qualitative project and constructed the interview guide. IH conducted the interviews. IH and ALM conducted the analysis and wrote up the first draft of the paper. All authors contributed to and have approved the final manuscript.


As the supply of starch grain and sugar cane, currently the main feedstocks for bioethanol production, become limited, lignocelluloses will be sought as alternative materials for bioethanol production. Production of cellulosic ethanol is still cost-inefficient because of the low final ethanol concentration and the addition of nutrients. We report the use of simultaneous saccharification and cofermentation (SSCF) of lignocellulosic residues from commercial furfural production  and corn kernels to compare different nutritional media. The final ethanol concentration, yield, number of live yeast cells, and yeast-cell death ratio were investigated to evaluate the effectiveness of integrating cellulosic and starch ethanol.Both the ethanol yield and number of live yeast cells increased with increasing corn-kernel concentration, whereas the yeast-cell death ratio decreased in SSCF of FR and corn kernels. An ethanol concentration of 73.1 g/L at 120 h, which corresponded to a 101.1% ethanol yield based on FR cellulose and corn starch, was obtained in SSCF of 7.5% FR and 14.5% corn kernels with mineral-salt medium. SSCF could simultaneously convert cellulose into ethanol from both corn kernels and FR, and SSCF ethanol yield was similar between the organic and mineral-salt media.Starch ethanol promotes cellulosic ethanol by providing important nutrients for fermentative organisms, and in turn cellulosic ethanol promotes starch ethanol by providing cellulosic enzymes that convert the cellulosic polysaccharides in starch materials into additional ethanol. It is feasible to produce ethanol in SSCF of FR and corn kernels with mineral-salt medium. It would be cost-efficient to produce ethanol in SSCF of high concentrations of water-insoluble solids of lignocellulosic materials and corn kernels. Compared with prehydrolysis and fed-batch strategy using lignocellulosic materials, addition of starch hydrolysates to cellulosic ethanol production is a more suitable method to improve the final ethanol concentration. Of the available biofuels that can partly replace the use of liquid petroleum to reduce greenhouse-gas pollution, ethanol is currently the most widely used . Starch Nevertheless, ethanol production from lignocellulosic materials, especially low-cost waste materials, is receiving increased attention . The utiallow disposal of waste products, reduce the landfill problem, and result in production of valuable products. Potential raw materials for bioethanol include wheat straw in Europe, corn stover in the USA and straw in China . Many inet al developed a novel starch-derived ethanol process using chemical and thermal treatment for conversion of non-starch polysaccharides in hulled barley into fermentable sugars [et al. [Much research has been devoted to reducing the cost of cellulosic ethanol by developing a low-cost pretreatment method, very efficient hydrolysis, and efficient fermentative microorganisms ,15,16. Ee sugars . A recene sugars . We confe sugars . However [et al.  showed tFurther reductions in cost would also expected from the reduction of chemicals used in the process. Addition of various nutrients is crucial for the efficient fermentation of cellulosic ethanol, and these contribute significantly to the cost of large-scale productions. Other studies on starch materials have shown that starch hydrolysates such as wheat hydrolysates are potential supplements for ethanol production from lignocellulosic hydrolysates .It does not seem economically feasible to hydrolyze cellulose and hemicellulose in starch materials by adding cellulase and β-glucosidase in starch ethanol-production processes; however, it might be feasible to add starch hydrolysates into the fermentation step using lignocelluloses as substrates. In this fermentation system, cellulase and β-glucosidase could then hydrolyze cellulose from both lignocellulosic and starch materials. Meanwhile, starch hydrolysates might be a source of some nutrients for yeasts, thereby decreasing the consumption of chemicals. Few studies have focused on the feasibility of such integration.In the present work, the simultaneous saccharification and cofermentation (SSCF) of FR and corn kernels, which have different nutrient compositions, were carried out. We investigated the final ethanol concentration, yield, number of live yeast cells, and yeast-cell death ratio. Compared with the fed-batch strategy and the use of prehydrolysis in lignocellulosic materials, the advantage of SSCF process was first tested in terms of the final ethanol concentration and yield. SSCF results from high water-insoluble solids (WIS) FR and corn kernels are summarized. Finally, the feasibility of ethanol production by SSCF of FR and corn kernels without additional organic nutrients was also evaluated.FR contains 42.2% cellulose, 38.7% lignin and 1.9% hemicellulose, and corn kernels contain 75.2% starch, 11.9% non-starch glucan and 17.7% washed solids residue (WSR). The concentration profiles of reducing sugars and ethanol during SSF/SSCF are shown in Figure SSF of lignocelluloses contains a lag phase because of the inhibition due to pretreatment byproducts and a change of medium. However, in our study, we did not see a lag phase in SSF of 5% FR, as the inhibitors had already been removed by the water-washing procedure. Ethanol production from SSCF of FR and corn kernels was faster than that from SSF of FR. The final ethanol concentration increased with increasing concentration of corn kernels in SSCF Figure , which iComparing the ethanol-yield profiles of SSF/SSCF Figure , the finThe final ethanol yield of SSCF increased with increasing concentration of corn kernels Figure . SSCF ofBy using a high concentration of substrates, a high final ethanol concentration was also obtained from cellulosic ethanol. However, a high concentration of lignocellulosic substrate (> 10%) would reduce the final ethanol yield because of higher viscosity and higher levels of inhibitor in the fermentation system . PrehydrFed-batch strategy is another potential method for performing SSF with high lignocellulose concentrations, because it can decrease the occurrence of stirring problems and the number of inhibitors in the fermentation system . HoweverProduction of a greater amount of ethanol from corn cellulose is an advantage of the integration of cellulosic and starch ethanol, and is partly responsible for the high yield obtained from SSCF of cellulosic and starch materials. Lignin is the second most abundant component of FR, and is a key challenge to cost-effective depolymerisation when using lignocellulosic materials. Notably, WSR is difficult to market as animal fodder because of the lignin accumulation that occurs in the cellulosic and corn ethanol integration strategy . High leThe number of live yeast cells increased in the first 17 h, and then decreased from 17 to 120 h Figure . The yeaDetoxification is always necessary for ethanol production from pretreated lignocelluloses to reduce the number of inhibitors . Severalet al. showed that SSF of softwood with 10 g/L yeast-cell mass had a higher overall ethanol yield (70%) than that with 2 g/L yeast-cell mass (64%) [7 yeast cells/ml reduced lactic-acid production significantly, while enhancing ethanol production [7 and 4 × 107 cells/ml [7 cells/ml. It is interesting that a high initial yeast-cell mass at the start of SSF can reduce lactic-acid formation and avoid excessive yeast reproduction, but also reduces the ethanol yield. According to Linde et al., in SSF of 7.5% steam-pretreated barley straw with 20 FPU/g cellulose, the ethanol decreased from 22.4 g/L to 21.4 g/L after 120 h, with a decrease from 5 to 2 g/L of yeast; however, ethanol concentration increased from 18.5 to 19.0 g/L with enzyme loading of 10 FPU/g [Lowering the yeast concentration can reduce the cost associated with yeast cultivation, because it is difficult to reuse yeast in SSF of lignocelluloses. However, a high yeast concentration may reduce lactic-acid production and enhance ethanol production. Stenberg ss (64%) . Other woduction . Low yeacells/ml . In our 10 FPU/g . Therefo10 FPU/g . We obtaSaccharomyces cerevisiae [The initial nitrogen concentration in the media strongly affects fermentation rate, but has no effect on the specific growth rates of revisiae . This corevisiae ,37, whicSSCF of cellulosic and starch ethanol can produce ethanol at a high concentration and yield. The yield of SSCF with both FR and corn kernels was higher than that of SSF with either FR or corn kernels alone, and the ethanol yield of SSCF increased with increasing concentration of corn kernels. The production of additional ethanol from corn cellulose is beneficial for the integration of cellulosic and starch materials, which leads to a high ethanol yield in SSCF.The integration of cellulosic ethanol with starch ethanol can also decrease chemical consumption. SSCF with mineral-salt medium produced the same ethanol yield as that with organic medium. The number of live yeast cells increased with increasing concentration of corn kernels in SSCF, while the yeast-cell death ratio decreased. Ethanol production from SSCF of a high substrate concentration of FR and corn kernels is therefore cost-efficient.Addition of corn hydrolysate increased the final ethanol concentration while maintaining high ethanol yield compared with SSF of a high level of lignocellulosic substrate with prehydrolysis. Compared with the fed-batch strategy for SSF or SSCF of the high WIS lignocellulosic substrate, SSCF of lignocellulosic substrates and starch materials together efficiently produced ethanol without a need for high enzyme loading or organic nutrients. In the starch ethanol-production system, integration of cellulosic and starch ethanol reduced the consumption of starch materials compared with the addition of cellulase and β-glucosidase.Lignin accumulation during the integration of cellulosic and starch ethanol makes it difficult to market the WSR as animal fodder, but studies on delignification could provide a potential way to overcome this disadvantage. More work is needed to evaluate the economic feasibility of the integration of FR and corn ethanol.Raw FR was kindly provided by Chunlei Company . Raw FR, with an initial pH of 2 to 3, was dried at 60°C for 12 hours after being rinsed with water to neutral pH. Washing procedures partly remove inhibitors, including furfural and 5-HMF . The corCorn-starch hydrolysis was performed in a 500-ml flask with 20% dry matter. Corn kernels were liquefied at 85°C for 2 h. Subsequent saccharification was performed at 60°C for 1 h (pH 4.0). The pH of the saccharification liquid was adjusted to 5.5 for SSF. Starch was liquefied and partially converted to glucose prior to performing SSF, to reduce the end-product inhibition of enzymes and the osmotic stress to yeast cells .In one experiment, corn kernels were treated as above for the liquefaction step, but the saccharification step was allowed to go for 24 hours to completely convert all starch to glucose. The glucose was measured to determine the total starch in the kernels, and the remaining WSR was isolated, dried, weighed and analyzed to determine the non-starch polysaccharide levels in the corn kernels.S. cerevisiae in the form of dry yeast . Dry yeast was activated in 2% glucose solution at 36°C for 15 minutes, then at 34°C for 1 hour. Thermostable Alpha-amylase (150 U/g corn kernels) and glucoamylase (20 U/g)  were used for corn liquefaction and saccharification, respectively. Cellulase /ml) and β-glucosidase  enzyme preparations  were used for SSF/SSCF; the amount of Celluclast 1.5 L and Novozyme 188 added was 15 FPU and 17 IU per gram cellulose of FR respectively. SSF of corn kernels was performed without Celluclast 1.5 L and Novozyme 188 supplements.The microorganism used for fermentation was 4)2HPO4 0.5 g/L, MgSO4 7H2O 0.5 g/L) and mineral-salt medium ((NH4)2HPO4 0.5 g/L, MgSO4 7H2O 0.5 g/L) were used. In each experiment, the conical flask was loaded with FR and fermentation medium, which were separately sterilized (121°C for 20 minutes). The enzymes, corn hydrolysate and yeast, with an initial cell mass concentration of 3.3 g dry matter per litre were then added to the conical flask directly. SSF/SSCF were performed at 38°C with an initial pH of 5.5. All experiments were agitated at 120 rpm in a shaking water-bath.The SSF/SSCF experiments were performed under nonsterile conditions in a 100-ml conical flask with a working weight of 60 g. In the anaerobic cultivations, each flask was equipped with a loop trap containing sterile glycerol. The concentrations ranged from 2.5% to 14.5% for corn kernels and from 5% to 7.5% for FR. Organic medium  to detect the reducing sugars and ethanol. The total amount of reducing sugars was measured by the Somogyi-Nelson colorimetric method, with glucose as a standard . EthanolSSCF: simultaneous saccharification and cofermentation; SSF: simultaneous saccharification and fermentation; FR: furfural residue (rinsed with water); WIS: water-insoluble solid; DDGS: dried distillers grains with solubles (a protein-rich byproduct of dry-mill starch to ethanol production); WSR: the washed solids residue (after complete saccharification of corn kernels).The authors declare that they have no competing interests.YT and JJ designed and coordinated the study, YT and DZ carried out the experiments, and YT and CC analyzed the results. YT and DZ wrote the paper, and CC and JJ reviewed the paper. All authors read and approved the final manuscript.


Sensors2012, 12, 16250–16261.” has been withdrawn at the request of the authors before the issue release of Sensors Volume 12, Issue 12. We apologize for any inconvenience this may cause.The following paper: “Jung, S.; Kim, J.H.; Kim, S. Bloom Filter-Based Advanced Traceback Scheme in Wireless Sensor Networks.


Of all the starch varieties studied, quinoa had the best overall emulsifying capacity, and OSA modified quinoa starch in particular. Although the size of the drops was relatively large, the drops themselves were in many instances extremely stable. In the cases where the system could stabilize droplets, even when they were so large that they were visible to the naked eye, they remained stable and the measured droplet sizes after 2 years of storage were essentially unchanged from the initial droplet size. This somewhat surprising result has been attributed to the thickness of the adsorbed starch layer providing steric stabilization. The starch particle-stabilized Pickering emulsion systems studied in this work has potential practical application such as being suitable for encapsulation of ingredients in food and pharmaceutical products.Starch granules are an interesting stabilizer candidate for food-grade Pickering emulsions. The stabilizing capacity of seven different intact starch granules for making oil-in-water emulsions has been the topic of this screening study. The starches were from quinoa; rice; maize; waxy varieties of rice, maize, and barley; and high-amylose maize. The starches were studied in their native state, heat treated, and modified by octenyl succinic anhydride (OSA). The effect of varying the continuous phase, both with and without salt in a phosphate buffer, was also studied. Quinoa, which had the smallest granule size, had the best capacity to stabilize oil drops, especially when the granules had been hydrophobically modified by heat treatment or by OSA. The average drop diameter ( In an emulsion, drops need to be stabilized in order to avoid coalescence. Surfactants adsorbed to the interface of the two phases decrease the interfacial tension and may increase steric hindrance or electrostatic repulsion, which increases the stability of the emulsion. Proteins and surfactants are usually used as emulsifiers in food emulsions. However, polysaccharides have also been used to stabilize emulsions, especially gum arabic, modified celluloses, and starches . When usOil drops stabilized by dispersed particles, known as Pickering emulsions, were originally observed independently by Starch granules naturally vary in size from 0.5 to 100 μm . DependiNative starch is not hydrophobic, and thereby generally not suitable to adsorb to the interface of water and oil and thus to stabilize an emulsion. However, by modification of starches, the hydrophobicity can be increased. Starch can be chemically modified by treatment with different alkenyl succinic anhydrides, for example, octenyl succinic anhydride (OSA), which is approved for food applications at an added amount of up to 3% based on the dry weight of starch. The hydrophobic octenyl group and the carboxyl or sodium carboxylate group increased starches' ability to stabilize emulsions . EmulsifIn our preliminary experiments, oil-in-water emulsions stabilized by OSA-modified quinoa granules were stable for more than 2 months and could be tailored to be creaming, sinking, or buoyancy neutral, depending on the starch to oil ratio . RheologThe objective of this study was to investigate the emulsifying capacity and storage stability of a broad spectrum of starches in their granular form. The starches used were both native and hydrophobically modified and had different granule size and amylose/amylopectin composition. The impact of salt concentration on the emulsifying capacity was studied in order to simulate the conditions of different food systems. Furthermore, the starch concentration was varied to analyze the effects on emulsion droplet size. The results of this study will be used to identify starches suitable for further emulsification and encapsulation studies.The following commercial starches have been investigated in this screening study: rice, waxy rice, maize, waxy maize, high-amylose maize (Hylon VII), and waxy barley . Starch isolated from quinoa grains  by wet-milling  has alsog for 10 min and washed twice with water and once with citric acid solution (pH 4.5) before the starch was air dried at room temperature for at least 48 h.Starch was thoroughly suspended in the double part by weight of water using a stainless-steel propeller and the pH was adjusted to 7.8. Four equal amounts of OSA  were added with an interval of 15 min and the pH was maintained at 7.4–7.9 by adding 1 mol/L NaOH solution drop by drop. When the pH was stable for at least 15 min, the starch solution was centrifuged at 3000g for 10 min, washed once with 50 mL ethanol (90%) and twice with water before the starch was suspended in 300 mL water, cooked in a boiling water bath for 10 min, and cooled to 25°C. The starch solution was titrated with 0.1 mol/L NaOH to pH 8.3. A blank was simultaneously titrated with native starch of the same origin as the OSA starch as a sample. The percentage of carboxyl groups from OSA on the starch granules was calculated byThe OSA substitution was determined by a titration method used in the industry (Lyckeby-Culinar AB). Briefly, 5 g (dry weight) of starch was dispersed in 50 mL 0.1 mol/L HCl and stirred for 30 min. The slurry was centrifuged at 3000where V is the volume (mL) of NaOH required for the sample and the blank titration, M is the molarity of NaOH (0.1 mol/L), W is the dry weight (mg) of the starch, and 210 is the molecular weight of octenyl succinate group.Dry starch was placed on glass dishes and heat treated in an oven at 120°C for 150 min in order to increase the hydrophobicity of starch granules and thereby achieve a higher affinity for a hydrophobic interface. This type of heat modification is based on work by The particle size distributions of the starch were measured using laser diffraction  in a flow through cell. The optical mode was Mie with a refractive index (RI) of starch of 1.54 . A smallThe starch granule samples were sputter coated with gold and images recorded in a scanning electron microscope  operated at 5 kV and a working distance at 8 mm. The lower electron imaging (LEI) detection mode was used as LEI gives a clear three-dimensional image of the surface of the samples studied. The LEI detector combines signals from both secondary electrons and of backscattered electrons.Emulsions were prepared in glass test tubes with 4 mL of the continuous phase, 2 mL of the oil phase, and 100–400 mg starch (50–200 mg/mL oil) by mixing with an Ystral  at 11,000 rpm for 30 sec. Approximately 1 mg of the oil-soluble dye Solvent Red 26 was added to the top of the emulsions after 24 h, and the test tubes were gently turned three times 1 h after addition in order to distribute the color in the entire sample. The color change in the emulsion was observed. The emulsions were shaken with a vortex mixer  for 5 sec after another 2 h to promote the interaction of the dyed oil with the rest of the emulsion drops. The color after vortex is a measure of the orthokinetic stability of the formed drops. Stable drops do not have an exchange with the lipophilic dye; hence, the emulsion phase will remain white. An increased red-colored emulsion phase indicates that the drops were less stabilized by the adsorbing starch granules or there is a free oil phase in the system. Afterward, the samples were stored at room temperature for 6 days before further analysis (described below). The samples were stored for another period of 2 years in sealed test tubes at room temperature before reanalyzing.For microscopy of the emulsions, an Olympus BX50 (Japan) microscope with Plan 2×, UMPlanFL 5× and 10×, and LMPlanFL 20× and 50× objectives (Olympus) was used. The light was transmitted using a polarization filter  and a wavelength filter . The emulsion was diluted five times with buffer corresponding to the continuous phase of the sample. The samples were placed on a microscopic slide, and the drops were studied immediately without cover glass. Images of the drops were taken with a 1280 × 960 resolution camera  connected to the microscope, and the images were processed using the Java image processing program ImageJ (version 1.42m).The test tubes with the emulsified samples were photographed 1 h, 24 h, and 6 days after the vortex treatment, and the images of the samples were analyzed in ImageJ. The emulsifying capacity of the starches and the stability of the emulsions were expressed as the volume of the emulsion to the total volume of the sample, often referred to as the emulsification index (EI). The amount of material, generally remaining starch, at the bottom of the test tube was also calculated. The EI  was calcd32) and the volume mean diameter (d43) were calculated using equations The drop size distribution of the emulsions was determined from microscopic images. The diameter of at least 250 drops was measured with ImageJ in the samples that contained drops that had a diameter smaller than 1.4 mm. The surface mean drop diameter  as percentage and the standard deviation have been calculated according to the following equations to arrive at the distribution of the emulsion drops in each sample.where After an additional 2 years of storage at room temperature (22°C) in glass test tubes, the starch granule-stabilized emulsions were photographed as described above. The starch granule-stabilized emulsions which had an initial mean particle size less than 120 μm determined by equation d32 of approximately 2 μm and had smooth edges  with 70% amylose was also used in order to see the emulsification behavior in a larger spectrum of the amylose content. It has been shown  that OSAd32 = 2 μm), had the preeminent best capacity to stabilize an emulsion at the circumstances used in this study. Native starch granules are supposed to be inefficient as oil drop stabilizers due to the low hydrophobicity; however, native quinoa and to some extent Hylon VII were able to stabilize the formed emulsion drops as seen in The color of the dyed emulsions is a measure of the interaction of the oil in the formed drops as the dye was added on top of the samples after the emulsification and before the samples were mixed in a vortex. Stable drops did not have an exchange with any dyed oil; hence, the emulsion phase remained white. An increased red-colored emulsion phase indicated that the drops were less stabilized by the adsorbing starch granules and that the interface barrier was more sparse, which lead to exchange of oil between the drops. The size of the drops correlated with the color and stability of the emulsion, that is, the more red the sample appeared the larger (or lack of) droplets there were in the system. This was mainly dependent on the size of the stabilizing granules, but also the shape of the granules had an impact. Quinoa, which had a unimodal drop size distribution of small granules  of the stabilized oil drops and the emulsifying index at equal starch to oil ratios. Quinoa had definitely best emulsifying capacity followed by rice, which only had slightly larger granule size, but the granules were more irregularly shaped with sharp edges. The emulsifying capacity was similar for the two varieties of OSA-modified rice  is reported in Table After storage at room temperature for a period of 2 years, the starch granule-stabilized emulsions were reexamined to see if they remain stable. Visual inspection of the test tubes showed that in all cases where there had been successful initial droplet stabilization, that is, in all the OSA-treated starches and some of the heat-treated starches, droplets remained and the emulsion layer had remained . This waTwo different phosphate buffers, with and without 0.2 mol/L NaCl, were used as continuous phase, and the pH was 7 in both buffers . Prelimid43 than the drops in the salt-containing emulsions, which had a more unimodal distribution. These observations can to a large extent be explained by the appearance of the drop systems shown in the microscopic images in When a continuous phase without salt was used, the emulsions had a distinct cone shape formed by the tip of test tubes during emulsification and remained during creaming (left images in d43) versus amount of added starch is plotted in The effect of starch concentration on emulsification was studied on four varieties of starch: quinoa, rice, maize, and waxy barley, all of which were OSA modified and used in a 0.2 mol/L NaCl phosphate buffer . These cThe samples with the highest amount of starch produced emulsions with a density higher than the continuous phase as shown in the sample with 200 mg starch/mL oil of waxy barley to the right in M, was estimated using equationTo quantify the effects of the amount of added starch and granule size, the maximum surface coverage possible for starch concentration with a given particle size was estimated. The assumptions made were that all starch particles were identical, spherical, and attached at the oil–water interface at a contact angle of 90° with an interfacial packing fraction φ ≍0.907, that is, hexagonal close packing . The thedsg is the surface mean diameter of the starch granule, ρsg is the starch density (1550 kg/m3), and φ is the packing density. Estimates of the maximum surface coverage, as well as the mean starch granule sizes for the various starches, are given in 2) increases with starch granule size, it is not surprising that the generated drop diameter in where The specific surface area of an emulsion, per volume of dispersed phase, is defined byd32 was measured by light scattering. Based on the amount of added starch, Csg (as mg per mL) and the theoretical maximum coverage, ΓM, of the given size of starch granules, a theoretical surface area that could be generated per volume of dispersed chase can be calculated, that is,where the surface mean diameter A comparison of the measured and calculated drop surface areas was plotted in By geometric analysis, it could be argued that as the ratio of starch granule size to forming drop size increases, the minimum surface coverage required to stabilize drops decreases, as larger spaces between the granules on the surface are possible while maintaining enough of a steric hinders to prevent coalescence. For this reason, the larger starch granules such as barley and maize had a larger surface area than predicted and the trend increased with increasing area . Microscope images confirmed this, showing larger spaces and gaps on the drops surface between adsorbed starch granules. In the case of rice, it had a smaller generated area than predicted (data points that lay below the line in Hydrophobic starch granules, especially when chemically modified, could be used to stabilize Pickering-type emulsions. The size of the granules had a large impact on the stabilizing capacity, and the smallest starch granules, from quinoa, had the predominantly best emulsifying properties. Also the shape of granules had slight impact on the emulsions. As expected, the emulsion droplet size decreased with increased starch concentration. The presence of salt in the continuous phase contributed to more uniform droplet sizes and less aggregation in the systems. The long-term stability of more than 2 years and the size of the emulsion drops formed make this type of starch granule-stabilized Pickering emulsions suitable for encapsulation of ingredients in food and pharmaceutical products.


Trudo Lemmens and colleagues discuss how the mere suggestion of litigation can bias the medical literature by affecting editorial decisions. They argue that journals and authors should publicly post threats of litigation, or cease and desist letters, for which there are some international legal precedents.Please see later in the article for the Editors' Summary The mere suggestion of litigation can bias the medical literature by affecting editorial decisions.Journals and authors should publicly post threats of litigation or cease and desist letters.There are some international legal precedents for publicly posting litigation threats.Posting of litigation threats has some advantages over commonly employed strategies for guarding against libel chill, such as legal consultations and litigation insurance.Editorial independence is critical if medical journals will continue to be venues for important debates that affect patient care. Editorial independence includes the latitude to accept or reject manuscripts on the basis of their content and without fear of repercussions against the journal. That independence is diminished when concerns about libel lawsuits deter journals from publishing contentious manuscripts. The Supreme Court of Canada has described this phenomenon of “libel chill” as follows: “There is concern that matters of public interest go unreported because publishers fear the ballooning cost and disruption of defending a defamation action. … When controversies erupt, statements of claim often follow as night follows day, not only in serious claims … but in actions launched simply for the purpose of intimidation. Of course ‘chilling’ false and defamatory speech is not a bad thing in itself, but chilling debate on matters of legitimate public interest raises issues of inappropriate censorship and self-censorship” Science's informal 2010 survey of 22 leading scientific and medical journals found that several journals “reported rejecting papers that were ‘clearly libelous’ or removing material that might have attracted libel action,” though “they insisted that this had been done on editorial grounds” New Zealand Medical Journal left a blank space in an article with the note: “the paragraph was withdrawn for legal reasons” It is difficult to determine how often concerns about litigation prevent manuscripts from being published or impact significantly on the content of manuscripts. In 1995, a Report of the Conference to Promote International Cooperation among Medical Journal Editors opined that “[e]ditorial freedom is, first and foremost, a freedom from a number of threats,” and that “[t]hreat of lawsuits against journals and editors seems to be an increasingly relevant and chilling factor in some countries” It is plausible that parties with large sums of money to gain or lose based on the content of a medical journal article  may attempt to exert their influence over editorial decisions with cease and desist letters or threats of litigation. This behavior can have a significant impact on editorial decisions, because of the power imbalance between a well-resourced and keenly motivated adversary and an often under-resourced journal with more submissions than can be published. Smaller journals may not be able to survive even the mere litigation process because of legal defense costs, as one of us was told by an editor when inquiring about the last minute pre-publication removal of a paragraph from an article published in the journal. While threats of lawsuits or subpoenas have been used to intimidate authors and even peer reviewers Science reported: “We know that on occasion academics may make assertions they can't then support; the legal advice we obtain often helps them to clarify their thinking and so we end up with a better paper, as well as one that should stand up in any court” A natural first response to a legal threat is to consult a lawyer. Journal editors are not generally professionally trained to deal with libel threats and even those with formal training may not have experience dealing with such threats. Obtaining such advice can improve the quality of a publication. As one scientific journal publisher quoted in Insurance may help journals defray the high cost of such legal advice. There are, however, two limitations to insurance schemes. The first is that insurance costs generally scale with the insured's perceived risk profile—in this case, its history and likelihood of attracting legal challenges. Journals publishing in controversial areas or willing to take a stance may be more likely to incur higher insurance costs. The second related problem is that insurance companies often exercise control over what approach to take when faced with legal action. They may insist on compromise or settlement, even when editors wish to press ahead with publication.Even if legal advice is affordable it may not be helpful to curb the impact of libel threats. While there are differences in how lawyers approach these threats, one reasonable approach for lawyers consulted by journals about manuscripts that can attract litigation is to err on the side of caution. They tend to have a professional interest in avoiding future criticism that could arise if a journal is taken to court following their more lenient legal advice. Cautionary advice, resulting in rejection of the manuscript, is perhaps more likely when legal consultation is mandated by a libel insurance policy. Decisions about publication may thus be affected by opinions from lawyers who are unable to comment on the scientific merit of the manuscript and who think foremost about the insurance company's interest in avoiding a lawsuit.We recommend that journal editors consider the option of publicly posting litigation threats, e.g., cease and desist letters or more subtle threats that are received either when a manuscript is being considered for publication or after it has been published.champion freedom of expression” The aim of this practice is deterrence and accountability rather than retaliation. Posting cease and desist or litigation threat letters will empower journals to publish worthy manuscripts despite threats and discourage specious claims meant to intimidate journals. The registry will expose such threats to scrutiny by the public, the government, and the press. Note that people or organizations will still be able to protect themselves against unwarranted allegations, as posting threats of litigation should not deter parties with legitimate concerns about being defamed from pursuing legal action. These should be able to publicly explain without embarrassment why they are pursuing legal action. In contrast, companies have an interest in avoiding exposure of their aggressive legal tactics if there is a weak basis for a threat. Making these threats transparent also promotes accountability of journal editors. A journal that refuses to publish an article after receiving a cease and desist or litigation threat letter can be pushed to explain its rationale. Posting threats promotes open discussion about freedom of speech by bringing to light a troubling aspect of research. It would, in our view, be a way to live up to what the Committee on Publication Ethics (COPE), an advisory body with a membership of over 8,000 journals, recognizes to be among the “[g]eneral responsibilities and duties of editors” to “This action can, of course, only work if many journals actually facilitate publication of these letters. The deterrent effect of the practice depends on its widespread adoption such that parties contemplating intimidation tactics actively take it into consideration. Organizations such as the International Committee on Medical Journal Editors (ICMJE), COPE, or the World Association of Medical Editors (WAME) could support this approach and promote adherence by making it a condition of membership. These organizations could also offer certifications to non-affiliated journals for their commitment to this approach, while these journals could also explicitly identify it as an essential component of their editorial policy. Furthermore, these organizations could take the lead in establishing and maintaining, with support of their members, a special website dedicated to this form of exposure of libel threats directed against medical journals. The advantage of the jointly organized website would be the ease of access for posting and for analyzing libel threats, the increased visibility of the initiative, and the sharing of maintenance costs and costs of initial legal consultations.Threat letters can be posted without comment on the merits of the threat and—where needed—with identifying information redacted. Editors should discuss threat letters and carefully assess how to publicize the letters in a way that squares with editorial integrity and risk management. Redaction of threatening letters to remove identifying information about the authors and the type of publication that is the target of a threat could be necessary if the decision is made not to publish a paper. The reason is that publicizing details about the rejected paper could undermine the anonymous submission and evaluation process of journals and could compromise the submission of the paper to another journal. In that case, an editorial note could accompany the publication of the letter, explaining in as much detail as possible the type of threat and the nature of the publication. An editorial note could also identify why the journal ultimately rejected the paper, whether it is because of the threat or for other editorial or quality assessment reasons. Of course, journals should consult legal counsel both before adopting any publication policy, as well as with respect to specific letters and threats.The specifics of a registry of threatening letters requires further elaboration with input of the editorial boards of leading medical journals. But it is worth pointing out here that there are precedents for this approach. One existing registry with a comparable objective, but focusing on internet postings and online publications—not on pharmaceutical and medical publications—is ChillingEffects.org, a website maintained by a coalition of American law school clinics and the Electronic Frontier Foundation Editors or those in charge of organizations that would take on the task of developing websites posting libel threats should carefully explore with expert counsel the legal consequences of publishing libel threat letters in all relevant jurisdictions, not just their own. Libel laws and other sources of liability have long arms such that what is safe under the laws of one jurisdiction may fall within the extraterritorial reach of another. We realize that this means that initially only journals or organizations with significant financial resources may feel comfortable to take the lead with this approach. Yet, there are promising indications and precedents in some Common Law jurisdictions that suggest it is a feasible approach that should not expose them to significant financial risks. We briefly discuss here selected examples from Canada and the United States as a discussion of implications in all countries is beyond the scope of this article.Angle v. LaPierrewithout comment on the letter itself or the underlying issue is not by itself defamatory.In Canada, a 2006 case from Alberta, Angle case dealt with a lengthy series of disputes between individual teachers and a teachers' association. The latter was suing the teachers for public statements concerning the disputes made on a number of public fora, including a website. The teachers had, among other things, republished a letter by the teachers' association in which it threatened legal action if they did not stop voicing their criticism in public, accompanied by the teachers' responses to those letters. The association challenged in particular public statements that referred to its tactics as legal or institutional bullying aimed at curtailing freedom of speech.The the conversation that ensued … was fair comment on the contents of the cease and desist letters. None of it was comment on any of the specific instances and events that prompted issuance of those letters” [our emphasis] While the court recognized that these accusations were “defamatory,” it ruled that several of them were protected as “fair comment,” since the legal tactic of the cease and desist letter could fairly be qualified as an attempt to “coerce a change in behaviour” any comment, and with private information redacted, would have been even more defensible. Of course, Angle also serves as a cautionary tale, for it makes quite clear that exchanges that ensue from the publication of letters could attract liability. Moreover, the Angle case is a lower court decision, and thus not binding on other courts. Yet it shows how the law of defamation has been applied favourably by at least one Common Law court to shield the publication of litigation threats.The court found that a forceful opinion stating that plaintiffs' letters “threatening legal action” constituted an attempt to silence opponents was protected as fair comment in part because it did not delve into the content of the controversy. By that same logic, republishing such letters without In the Unites States, which for several decades has had what some view as “the most speech protective substantive libel laws in the world,” Media law scholar Sam Bayard at Harvard University's Berkman Center for Internet and Society concludes his discussion of the issue by stating: “Don't be bullied by a lawyer threatening you with a copyright infringement suit [under American law] for republishing the contents of a threatening letter. One way or another, this is an extremely weak legal argument, and one that the lawyer is extraordinarily unlikely to pursue” There is also the possibility that attempts to sue those who post litigation threats would actually result in greater public scrutiny of the larger issue of “libel chill” and could result in changes to the law, just as high profile defamation cases in the United Kingdom led to changes in the law that now protect important scientific debate Medical journals should explore the option of bolstering their commitment to editorial independence by publicly posting threats of litigation. Consultations with lawyers and litigation insurance may reduce the likelihood of lawsuits but neither insulates editorial decisions from libel chill. While exposing litigation threats will not completely prevent them, it may be a step towards exposing the problem, increasing accountability, and fostering new social, scholarly, and legal norms.


PLOS Computational Biology is the Public Library of Science journal that targets new biology that is facilitated by computational methods. Since the inception of the journal, biology has been at the center of the scope of PLOS Computational Biology. Thus, each submission is required to put advances in biology into its focus in a very concrete manner and not only focus on computation. This is in contrast to other, more technically oriented journals in the area of computational biology.PLOS Computational Biology has installed a special type of submission, the Methods paper. As the scope statement of the journal spells out, “Methods papers should describe outstanding methods of exceptional importance that have been shown, or have the promise to provide new biological insights. The method must already be widely adopted, or have the promise of wide adoption by a broad community of users. Enhancements to existing published methods will only be considered if those enhancements bring exceptional new capabilities.”The new Methods section of the journal acknowledges the fact that a major methodical advance can, in itself, be so relevant that it deserves transcending the technological domain and being presented to a broader readership including not only computational biologists, but also biologists, the targeted readership of this journal. For this reason, In order to render the processing of Methods papers as effective as possible and to limit the effort required on the part of authors and reviewers, a presubmission inquiry is mandatory for Methods papers. In such an inquiry, the authors are requested to present a concise abstract-like statement on what the manuscript they plan to submit entails and why the authors feel that it fits the Methods section of the journal. Presubmission inquiries are given top priority in the paper handling process; the median processing time should be about a week. Within that time, submission is either encouraged or discouraged.The Methods section of the journal is handled by Deputy Editor Thomas Lengauer.


Lepidoptera encompasses more than 160,000 described species that have been classified into 45–48 superfamilies. The previously determined Lepidoptera mitochondrial genomes (mitogenomes) are limited to six superfamilies of the lineage Ditrysia. Compared with the ancestral insect gene order, these mitogenomes all contain a tRNA rearrangement. To gain new insights into Lepidoptera mitogenome evolution, we sequenced the mitogenomes of two ghost moths that belong to the non-ditrysian lineage Hepialoidea and conducted a comparative mitogenomic analysis across Lepidoptera.Thitarodes renzhiensis and T. yunnanensis are 16,173 bp and 15,816 bp long with an A + T content of 81.28 % and 82.34 %, respectively. Both mitogenomes include 13 protein-coding genes, 22 transfer RNA genes, 2 ribosomal RNA genes, and the A + T-rich region. Different tandem repeats in the A + T-rich region mainly account for the size difference between the two mitogenomes. All the protein-coding genes start with typical mitochondrial initiation codons, except for cox1 (CGA) and nad1 (TTG) in both mitogenomes. The anticodon of trnS(AGN) in T. renzhiensis and T. yunnanensis is UCU instead of the mostly used GCU in other sequenced Lepidoptera mitogenomes. The 1,584-bp sequence from rrnS to nad2 was also determined for an unspecified ghost moth (Thitarodes sp.), which has no repetitive sequence in the A + T-rich region. All three Thitarodes species possess the ancestral gene order with trnI-trnQ-trnM located between the A + T-rich region and nad2, which is different from the gene order trnM-trnI-trnQ in all previously sequenced Lepidoptera species. The formerly identified conserved elements of Lepidoptera mitogenomes (i.e. the motif ‘ATAGA’ and poly-T stretch in the A + T-rich region and the long intergenic spacer upstream of nad2) are absent in the Thitarodes mitogenomes.The mitogenomes of T. renzhiensis and T. yunnanensis exhibit unusual features compared with the previously determined Lepidoptera mitogenomes. Their ancestral gene order indicates that the tRNA rearrangement event(s) likely occurred after Hepialoidea diverged from other lepidopteran lineages. Characterization of the two ghost moth mitogenomes has enriched our knowledge of Lepidoptera mitogenomes and contributed to our understanding of the mechanisms underlying mitogenome evolution, especially gene rearrangements.The mitogenomes of  Drosophila yakuba mitogenome, the first insect mitogenome to have its sequence determined, is shared by the majority of insect species, and this is therefore considered to be the ancestral order for the entire class Insecta  and T. yunnanensis [GenBank: HM744695]). Because of expansion of the A + T-rich region, the mitogenome sequence of T. renzhiensis is longer than any other complete Lepidoptera mitogenome sequenced to date. The 1,584-bp mtDNA sequence comprising partial rrnS, the A + T-rich region, trnI, trnQ, trnM, and partial nad2, was also determined for Thitarodes sp. [GenBank: HQ883371].The complete mitogenomes of T. renzhiensis and T. yunnanensis mitogenomes both include the entire set of 37 genes 3CTTTTTCAACTA whereas there is no such intergenic spacer in T. renzhiensis. There is a 15-bp intergenic spacer between trnS(UCN) and nad1 in both mitogenomes. In this region, a conserved motif ATACTAA is present in all previously sequenced Lepidoptera mitogenomes , AUU (Ile), and UUU (Phe). As a consequence, Leu (UUR), Ile, and Phe are the three most frequently used codon families  and one truncated repeat unit (101 bp). However, the A + T-rich region of Thitarodes sp. consists entirely of non-repetitive sequences  [Papilio maraho (252 bp) [Spilonota lechriaspis (124 bp) [The A + T-rich region of (126 bp) , Papilio(252 bp) , and Spi(124 bp) ,59. ThesrrnS gene in the previously sequenced Lepidoptera mitogenomes, there is a widely conserved structure that includes the motif ‘ATAGA’ and a 16–22 bp poly-T stretch. It has been suggested that this structure might function as a signal for mtDNA replication initiation [T. yunnanensisT. renzhiensis, and Thitarodes sp., indicating that these structural motifs are not conserved in the non-ditrysian Lepidoptera species. The Thitarodes mitogenomes may adopt a different strategy for replication initiation. The absence of the conserved elements in the A + T-rich region, together with above mentioned structures, such as the presence of the ancestral gene arrangement, and the absence of the intergenic spacer upstream of nad2, demonstrates that the Thitarodes mitogenomes have unusual features compared with the previously determined Lepidoptera mitogenomes. Therefore, in future studies, more attention should be paid to non-ditrysian lineages when comparing Lepidoptera mitogenomes.Downstream of the itiation ,60,61. HThitarodes renzhiensis and T. yunnanensis mitogenomes are the first representatives of non-ditrysian lineages of Lepidoptera. The arrangement of the tRNA genes between the A + T-region and nad2 is trnI-trnQ-trnM, different from those of previously sequenced Lepidoptera mitogenomes but identical to the ancestral gene order. Therefore, the previously identified tRNA rearrangement is not a synapomorphy for Lepidoptera mitogenomes. This result indicates that the tRNA rearrangement event(s) likely occurred after Hepialoidea diverged from other lepidopteran lineages. In addition, compared with other Lepidoptera mitogenomes, T. renzhiensis and T. yunnanensis mitogenomes have other unique structural characters such as the lack of the intergenic spacer upstream of nad2. Therefore, non-ditrysian lineages should be taken into consideration in future comparative mitogenomic studies of Lepidoptera. Characterization of the two mitogenomes has contributed to our understanding of Lepidoptera mitogenomes and provided insights into mitogenome evolution, especially gene rearrangements.T. renzhiensis and T. yunnanensis were obtained from cultures in Diqing, Yunnan Province, China. Specimens of Thitarodes sp., whose complete binomial name has not been specified, were collected in the Qilian Mountain, Qinghai Province, China. All the specimens were preserved in anhydrous ethanol and stored at −20 °C until used for DNA extraction. Whole genomic DNA was extracted from an ethanol-preserved larva with the DNeasy Blood & Tissue kit . DNA quality was assessed by electrophoresis in a 1 % agarose gel stained with ethidium bromide.Samples of T. renzhiensis and T. yunnanensis were amplified in 10 overlapping fragments (F1 to F10) using long PCR. All primers were designed based on the conserved nucleotide sequences of the known Lepidoptera mitogenome sequences (Table The entire mitogenomes of both es Table . Primer Raw sequence files were proof read and assembled into contigs using ContigExpress included in the Vector NTI (9.1) package.PCGs were determined and translated into putative proteins using the ORF finder in the DNAStar package . The identity of these polypeptides was established using the BLAST program available on the NCBI web site. To determine the gene boundaries, the 5' ends of PCGs were assumed to be at the first legitimate in-frame start codon  in an open reading frame (ORF) that was not located within an upstream gene encoded on the same strand; the 3' ends were inferred to be at the first in-frame stop codon encountered downstream of the start codon. When the stop codon was located within the sequence of a downstream gene encoded on the same strand, a truncated stop codon (T or TA) adjacent to the beginning of the downstream gene was designated as the termination codon ,11. ThisrrnL was annotated to extend to boundaries of the flanking trnL(CUN) and trnV. The 3' end of rrnS was annotated to be adjacent to the start of trnV, while the 5' end was determined via comparison with orthologous sequences of other Lepidoptera mitogenomes using MEGA ver4.0 [Identification of tRNA genes was performed using the tRNAscan-SE Search Server  with invA ver4.0 .http://amiga.cbmeg.unicamp.br/) [T. renzhiensis and T. yunnanensis mitogenomes were calculated with MEGA. To measure synonymous codon usage bias, RSCU values were also computed. Average codon usage and RSCU values were further analyzed for each superfamily used in this study. GC-skew = (G-C)/(G + C) and AT-skew = (A-T)/(A + T) were used to measure base compositional differences [Nucleotide sequences of the 13 mitochondrial PCGs of the previously determined 37 Lepidoptera species (Table amp.br/) . Nucleotferences . The tanferences .atp6 and atp8, ATP synthase subunits 6 and 8; cob, cytochrome b; cox1–3, cytochrome c oxidase subunits 1–3; nad1–6 and nad4L, NADH dehydrogenase subunits 1–6 and 4 L; rrnS and rrnL, small and large ribosomal RNA (rRNA) subunits; trnX, transfer RNA (tRNA) genes with X representing the one-letter abbreviation of the corresponding amino acid.The authors declare that they have no competing interests.DRY conceived and designed this study. JYC collected specimens and extracted DNA. YQC carried out molecular experiments. YQC and CM analyzed the data and drafted the manuscript. DRY thoroughly revised the manuscript. All authors contributed to the manuscript and approved the final version.Figure S1. Codon distribution in mitogenomes of currently used lepidopteran superfamilies. CDspT, codons per thousand codons.Click here for fileFigure S2. The relative synonymous codon usage (RSCU) in mitogenomes of currently used lepidopteran superfamilies.Click here for fileTable S1. PCR primer sequences.Click here for file