The discovery of penicillin transformed the medicine industry and initiated a search for a better antibiotic every time resulting in several synthetic and semi-synthetic antibiotics. Beginning with the 1937 sulfa drug tragedy, the drug regulations had a parallel growth along with the antibiotics and the antibiotic-based generic Pharma industries. This review article is focused on the scenario depicting current global Pharma industries based on generic antibiotics. Several regulatory aspects involved with these industries have been discussed along with the complexity of the market, issues that could affect their growth, their struggle for quality, and their compliance with the tightened regulations. With the skyrocketing commercialization of antibiotics through generics and the leveraging technologic renaissance, generic industries are involved in providing maximum safer benefits for the welfare of the people, highlighting its need today..Ever since the discovery of antibiotics, the quality of human life greatly improved in the 20 Humans are constantly exposed to a variety of microorganisms, including the major groups, namely, bacteria, fungi (yeasts and molds), algae, protozoa, and viruses. Despite steadily improving public awareness and continued diagnostic and therapeutic advances, human race faces a continuous threat from microorganisms through infections. Many individuals develop a variety of infections but quickly overcome them. In most cases these microorganisms do not produce infection because the skin and mucous membrane surfaces provide effective barriers against invasion. However, some individuals are unfortunate as a few microorganisms can invade through the barrier or through lesions from surgery or trauma and develop chronic or persistent infections. Statistics show 2.5 deaths on an average per 1 million people in the USA as a result of bacterial infections.[At the beginning of the 20th century, a metabolic approach was applied to the formulation of new drugs. German bacteriologist Paul Ehrlich contended that since various cells of the body and of various microorganisms could be selectively stained by certain dyes, there must be specific active groups in cells of the human body and in microorganisms to which drugs of the dye type might attach. Such a drug would then act as a “magic bullet,” attacking the target cells specifically and killing only the microorganisms, while leaving the human body unaffected. Drug devThe pharmaceutical industry develops and produces a variety of medicinal products that save the lives of millions of people from various diseases and permits many people suffering from illness to recover and to lead productive lives. There are different types of pharmaceutical companies, namely, mainline pharmaceutical companies, which are established firms that have many approved drugs already on the market. These companies often have a significant number of research and development (R&D) laboratories and manufacturing plants throughout the nation and around the world. In contrast, smaller pharmaceutical companies often do not have any approved drugs on the market. In addition to developing their own drugs, some of them may perform contract research for other pharmaceutical companies. Finally, generic pharmaceutical companies manufacture drugs that are no longer protected by patents. Because their products are all established drugs, they devote fewer resources to R&D and more to manufacturing.A generic medicine is a faithful copy of a mature drug—no longer under patent—marketed with the chemical name of the active ingredient. It is a pharmaceutical product intended to be bioequivalent with the innovator or the company which launched the new drug, manufactured without a license from the innovating company and marketed after expiry of a patent or other exclusivity rights. The development of generics markets stems from at least 2 major points. On the demand side, to face the rapid growth of health care expenditures, health care systems have been reformed recently. On the supply side, however, in this decade the slow input of innovative drugs and the expiry of patents on many important medical specialties have led to increasing price competition, favoring generics in many developed countries.The generic industries play a major role in the cost reduction of pharmaceutical drugs in general and antibiotics in particular. Historically, the pharmaceutical industry capitalized on the discovery that many microbial secondary metabolites act as antibiotics. Even today, significant portion of the worldwide pharmaceuticals are dedicated to the production of antibiotics. Prescription drugs represent a significant component of increasing costs, with shares ranging from 4% in the USA to nearly 18% in France and Italy. The indu8The USA market is considered to be one of the biggest markets in the health care sector due to its million dollar business potential and is the primary target for most of the generic pharmaceutical manufacturers. In USA, the pharmaceutical industries have achieved worldwide reputation through R&D on new drugs and spend a relatively high amount of its profits on R&D compared with other industries. Each year, pharmaceutical industry testing involves millions of compounds, yet in the long run, yields only fewer new fruitful medicines. It is critical to maintain appropriate incentives for the development of new drug products, because the necessary R&D is risky and costly. At the same time, expenditures on pharmaceutical products continue to grow and often outpace expenditures for other consumer products. The new drug development, as well as the generic drug availability is well balanced by the current regulations in the USA through implementing the Hatch–Waxman Act.In the USA, oral and parenteral antibiotics were found to possess equal shares of sales. The estimates of antibiotic availability from numerous countries throughout the world serve as an indicator of potential patterns of human antibiotics sales. The extraordinary therapeutic effects of antibiotics, the occurrence of resistance, and the considerable resources spent on antibiotics worldwide were the compelling reasons for concern about adequate and appropriate use of these powerful agents. Antibiotics often accounted for 15%–30% of drug expenditures, the largest share of expenditure among any therapeutic group of drugs. According to the US Centers for Disease Control and Prevention, 1.7 million people per year in the USA face hospital-acquired infections, leading to 5.8% deaths among them. Recent fThe FDA or USFDA is an agency of the US Department of Health and Human Services, responsible for protecting and promoting public health through the regulation and supervision of prescription and over-the-counter pharmaceutical drugs, vaccines and biopharmaceuticals, and many other commodities, which may influence public health. FDA plays an inevitable role in the approval of generic as well as new drugs, which are to be marketed in the USA. The Drug Price Competition and Patent Term Restoration Act of 1984, usually referred to as the Hatch–Waxman Act, was designed to promote generics in the USA while leaving intact a financial incentive for R&D. It allows generics to win FDA marketing approval by submitting bioequivalence studies. Approvals were generally provided with the following certifications:Paragraph I Certification: The generic applicant certifies that there are no patents listed in the orange book. “Orange book” being a publication of USFDA, lists the patents relating to drugs approved for marketing and sale in the USA, including patents that protect active ingredients.Paragraph II Certification: In case any listed patents have previously expired, the applicant may enter the marketplace immediately upon FDA approval.Paragraph III Certification: The applicant certifies that any listed patent has not yet expired but will expire on a particular date. The FDA may approve the Abbreviated New Drug Application (ANDA) and make it effective as of the patent expiration date.Paragraph IV Certification: The applicant for generic approval intends to market the drug prior to expiration of any patent(s) listed in the orange book; the applicant makes a certification that the patent(s) are not infringed or are invalid and FDA notifies the New Drug Application (NDA) holder and patent owner accordingly. It also Another provision of the Hatch–Waxman Act is that it grants a 30-month stay to drug companies that file suits against generic manufacturers who challene their patents. Thus the act maintains a fair balance between the innovator of a new drug and the generic drug producers.The history of antibiotic regulation clarifies the relationship between regulatory plan and the scientific/regulatory constraints and the marketing conditions in which they operate. Antibiotics and insulin-containing drugs were added to the regulatory scheme beginning with a series of steps in 1941. However, the procedures for establishing safety and efficacy applicable to other “new drug” antibiotics were subject to a far different regulatory scheme.In the case of antibiotics, the monographs were developed on the basis of the first product or the innovator product reviewed and approved in the antibiotic class. Thereafter, any forthcoming vendor merely needed to show that it was bioequivalent to the innovator product for which the monograph was developed and that it followed the specifications of the monograph. In this way, the innovator owned no rights even though the following vendors receive the approval based on his certificate. This seems to be a paradox.Along with this, the antibiotic vendor should provide a sample of each batch of the antibiotics to the FDA for laboratory testing and certification. The batches were tested by the agency and if found to meet the standards, the Antibiotic Certificate was issued. Also, thIn 1986, over-the-counter antibiotics that complied with the applicable monograph were excluded from the batch certification process. In contrast to the earlier times where only penicillin was the available antibiotic in the market, several hundreds of antibiotics started getting approval from the agency. As a result of the 1962 Amendments, the FDA required the submission for several antibiotics of scientific evidence of substantial well-controlled clinical studies, demonstrating the effectiveness of the product. Those products that failed to provide such evidence had their certifications overturned. In addition, the FDA cancelled approval of several antibiotics that did not have substantial scientific evidence. In 1985, antibiotic applications were classed as either “New Antibiotic Drug Applications” or “Abbreviated Antibiotic Drug Applications.” In line with the new regulatory framework, applicants and manufacturers could now make certain changes in their products without requiring prior FDA approval.In 1997, antibiotics were started to be reviewed and approved on the same basis as any other pharmaceutical product. In 2000, the FDA recounted the term “antibiotic drug” as it refers not only to the active chemical substance, but to any derivative of the substance, such as a salt or an ester of the substance. Under the new scheme, similar to nonantibiotics, antibiotics could also henceforth contain drug substance information in the finished product application itself. Thus, antibiotics entered into the normal stream of pharmaceuticals as any other pharmaceutical categories.The antibiotics market generated sales of US$42 billion in 2009 globally, with 14 products recording sales of more than $1 billion.18 There Japan is the second-largest pharmaceutical market in the world. It is well known that Japan is the third country next to the USA and the UK to become self-sufficient in penicillin manufacture as early as in 1948. Besides this, much effort was made in exploratory research on anti-infectives. Starting from colistin, several antibiotics, namely, kanamycin, bleomycin, piperacillin, norfloxacin, meropenem, and others, are from Japan. As of the 12 months to Q1 2009, the Japanese pharmaceutical market accounted for nearly 10% of the world market, and generated total sales of $71.6 billion. The Japanese government ensures that all citizens and workers have health insurance. Compared with that in 2008, Japan tightened monitoring and inspection of imported antibiotics in 2009 with more stringent requirements for their testing.Several regulatory challenges were involved in the development of a generic antibiotic. Regulatory challenges, such as bioequivalence, patent expiry, newer antibiotic, and the complexity involved in the regulated market, are explained in the following sections. max), and the time to peak concentration (tmax). Statistically, geometric mean ratio of the test to the reference drug for AUC and Cmax must fall within 90% confidence limits of 80 and 125.[In order to protect consumers, generic products must be demonstrated to be therapeutically equivalent to a previously approved product, typically an innovator product. Nowadays bioequivalence studies are essential part of a company’s registration dossier. These bioequivalence studies measure the bioavailability of 2 formulations of the same active ingredient. The plasma concentration of a drug determines the number of drug molecules at a receptor and hence the therapeutic effect. The plasma concentration is governed by absorption, distribution, metabolism, and elimination. The pharmacokinetic parameters measured are area-under-the-concentration (AUC) time curve, the peak concentration (C and 125.21 WithinSubstitution of generic drugs for brand name products is highly controversial, especially in antibiotics and often is met with suspicion by health care providers and patients. Bioequivalence issues present more concerns over generic drug substitution, such as consumer perception of risk, differences in product and packaging appearance, and differences in excipients. Also, several antibiotics, such as cephalosporins, have relatively narrow therapeutic window and would undergo stringent bioequivalence testing, because relatively modest changes in the concentration achieved in body fluids might well be associated with large changes in the frequency of therapeutic failure or significant toxicity. It arises several questions as whether is it possible to market various intravenous products with same concentration of active ingredient but with variable quality, even if it does not affect the clinical outcome, it does little to build trust in the generic industry.As the earth moves closer to the concept of a global village by elimination of trade barriers, new challenges crop up. A case in point is the market for generic antibiotics. Hatch–Waxman established a regulatory framework that sought to balance incentives for continued innovation by brand name companies and opportunities for market entry by generic drugs. Before 1962, it was observed in the USA that out of the 150 off patent drugs in the market, there were no generic drugs. Many companies did not go in for manufacture of generic drugs because of the impractical and nonscientific manner in which the regulatory authorities viewed the approval process. The Hatch–Waxman Act addressed these issues and proposed many reforms. The underlying objective of the Act is that in the absence of generic drugs, it is difficult to check the profiteering motive of the patent owner of a drug, who may put patients to ransom. However, at the same time it also takes care of the interest of the patent owner and provides relief for undue lengthy process. It is now possible for many generic companies to qualify for the 180-day market exclusivity if several applications are filed on the same day. Under the Hatch–Waxman Act, the government has a system of patent term “restorations” under which monopoly of the original patentee can be extended for a maximum period of 5 years in addition to the initial patent term. In the European Union also there exists a system of supplementary protection.Although the need for new antibiotics is increasing, a number of factors make these drugs less economically attractive than drugs that treat chronic diseases. Pharmaceutical companies appear to be less interested in developing anti-infective drugs. Reason b26Even though it looks simpler, the markets for generic antibiotics are more complex due to several factors. Several factors that influence the generic antibiotic market growth are depicted in Streptococcus pyogenes, cutaneous staphylococci, and propionibacteria. In a hospital setting, an increased use of antibiotics is often associated with an increase in the frequency of antibiotic resistance. Furthermore, the relationship between antibiotic use and resistance is most evident when resistance is due to mutations selected during therapy, resulting in clinical failure. The inappropriate use or overuse of antibiotics has caused the major problem of polydrug resistance, and resistance patterns of bacteria to the new antibiotics are likely to parallel the extent to which they prescribe. Such general antibiotic resistance affects the market to a greater extent.[The evolution of antibacterial resistance in human pathogenic microorganisms is the result of the interaction between antibiotic exposure and the transmission of resistance within and between individuals. In the community, there is cumulative evidence that the antibiotic exposure of populations promotes acquired antimicrobial resistance in community pathogens, such as r extent. In todayIn the USA, a black box warning is a type of warning that appears on the package insert for prescription drugs that may cause serious adverse effects. It is so named for the black border that usually surrounds the text of the warning. Several antibiotics, including ciprofloxacin, gemifloxacin, levofloxacin, moxifloxacin, norfloxacin, and ofloxacin are affected by these blackbox warnings. Similarly “Dear Doctor” letter is intended to alert physicians to safety precautions that should be taken to reduce the potential risk reported to be associated with the drug products. Even though these warnings may be common for both the generic and branded drugs, these label warnings may create a negative impact on the public perception with the generic antibiotics. Apart from these, drug withdrawal and recall procedures affect the generic market in varying degrees. Recalls are an appropriate alternative method for removing or correcting marketed consumer products, their labeling, and/or promotional literature that violate the laws administered by the FDA. Recalls Pharmaceutical firms that produce an antibiotic are usually given temporary monopoly power through a patent, granted to recover the incurred investment in R&D and by this to encourage future innovation of new drugs. The granting of this monopoly power ignores the fact that this also gives them some control over the levels of the drug’s treatment efficacy on the one hand, as well as of the infected population on the other. In turn, a too intensive use of antibiotics within the community may lead to an increase in the bacterial resistance of the drug. Furthermore, literature from the USA has shown that brand name manufacturers do not compete on price once generic competitors become available. The lack of price competition may lead to increased costs in the private market. Private insurance companies generally do not require generic substitution and some provinces do not require generic substitution for cash-paying customers. FurthermAntibiotics are among the most frequently prescribed medications in modern medicine. Most antibiotics have 2 names, the trade or brand name, created by the drug company that manufactures the drug or otherwise known as innovator, and a generic name, based on the antibiotic’s chemical structure or chemical class. The antibiotic market has evolved itself to a much stable place in spite of its complexity. The regulations of global health communities or agencies have been strictly developed on these antibiotics due to their potential in curing life-threatening diseases. Previously, a special regulatory care was given to antibiotics rather than any other drugs due to their relatively newness and also due to their significance in usage by public for life-saving purpose. Antibiotics were subjected to a batch certification requirement apart from complying with the monograph. This regulation required that each batch of antibiotic produced be certified to conform to the regulations of identity, strength, quality, and purity before marketing in the USA. This resulted in an aggressive cost increase in the antibiotic sector as well as time delay for approval. Nowadays, after several amendments to the regulations by the FDA, antibiotics are being regulated as any other pharmaceuticals. At the same time, industries are not showing interest in discovering newer antibiotics due to several factors, such as drug resistance, cost involved in the research, comparatively less profit due to industrial competencies, and so on. The introduction of Hatch–Waxman Act has made a paradigm shift in the industry by balancing the benefits for both the innovator and the generic industry. The introduction of only few newer antibiotics in the last 5 years shows the lack of awareness about these antibiotics. Although the infections were inhibited when improved sanitation and drugs were applied to combat microbes, and while many of the terrifying acute diseases, such as typhoid, cholera, and dysentery, have been subdued, many serious microbial diseases have not been eliminated. With the dangerously growing levels of antibiotic resistance and newer type of infections, organisms, and so on, it emphasizes the need for a more streamlined and enhanced means of developing and approving new agents, the need for greater integration of oversight and policy development efforts, and the necessity of greater availability of better data. Rather, regulations should be tightened up to avoid improper prescriptions and the misuse of these antibiotics.However, the introduction of Hatch–Waxman Act has in fact increased the generic antibiotic industries and has a definite effect on their growth. The law requires that these drugs must meet the specifications of the official compendia, as any other pharmaceuticals. Although several efforts have been taken by the regulatory agencies devoted to the tests and methods of assay of antibiotics, including sterility, biological test, microbiological and chemical assays, general and specific chemical tests, and tests on specific dosage forms, the generic industries struggle hard to bring the quality and safe antibiotics complying with these regulations. In future, in spite of the complexity and competency in the field, the wellbeing of the generic as well as the industries involved with the new drug inventions should be well protected to preserve the public health.In this article, generic Pharma industries with respect to antibiotics were explored. Considering the Hatch–Waxman Act, how the generic approvals made as fair has been explained. Also, history of antibiotics, its regulations, current market scenario, and several regulatory challenges facing antibiotic industries have also been discussed with an effort to bring into light a specialized field with challenges. However, with the increase in globalization and the demand for newer antibiotics, industries should be encouraged with focus on the future regarding the antibiotic. In summary, it is critical to devise strategies for antibiotic regulations, to ensure public health with wellbalanced commercial and regulatory aspects.


Seizures are a common problem in small animal neurology and it may be related to underlying diseases. Porencephaly is an extremely rare disorder, and in Veterinary Medicine it affects more often ruminants, with only few reports in dogs.A one-year-old intact male Shih-Tzu dog was referred to Veterinary University Hospital with history of abnormal gait and generalized tonic-clonic seizures. Signs included hypermetria, abnormal nystagmus and increased myotatic reflexes. At necropsy, during the brain analysis, a cleft was observed in the left parietal and occipital lobes, creating a communication between the subarachnoid space and the left lateral ventricle, consistent with porencephaly; and also a focal atrophy of the caudal paravermal and vermal portions of the cerebellum. Furthermore, the histological examination showed cortical and cerebellar neuronal dysplasia.per se or in assemblage could result in seizure manifestation.Reports of seizures due to porencephaly are rare in dogs. In this case, the dog presented a group of brain abnormalities which  Seizures are a common problem in small animal neurology and it may be related to underlying diseases. Brain anomaly associated with seizures was described in 4.16% of 240 dogs in a retrospective study, hThe main clinical manifestation of porencephaly is the occurrence of seizures,14, wherIn this case, CSF evaluation was normal. The dog had anesthetic complications without any apparent cause and died, and according to Gaynor et al., it is aThe major gross alteration presented herein is consistent with porencephaly. We also found cerebral neuronal dysplasia and cerebellar vermis focal atrophy. Even though porencephaly has been associated with viral infections and nutritional deficit in ruminants and vascular disturbs in humans, the specific cause in dogs remains undetermined,15. ViraPorencephaly is an extremely rare disorder of the brain, with no previous description in Shih-Tzu dogs, and this report gives additional evidences to relate porencephaly to the occurrence of seizures. Therefore, as observed by Davies et al. as well Orally informed consent was obtained from the owner of the dog for publication of this case report.None of the authors of this paper have any financial or personal relationship with other people or organizations that could inappropriately influence or bias the content of this paper.GFM is the supervisor responsible for the case report and was responsible for collecting samples and interpretation of the histological findings. MGL participated in clinical research. AS and GDM performed the analysis and interpretation of the histological findings and photographed the images. All the authors helped to draft the manuscript. All authors read and approved the final manuscript.


New theoretical and conceptual frameworks are required for evolutionary biology to capitalize on the wealth of data now becoming available from the study of genomes, phenotypes, and organisms - including humans - in their natural environments. We live in an exciting time for biology. Technological advances have made data collection easier and cheaper than we could ever have imagined just 10 years ago. We can now synthesize and analyze large data sets containing genomes, transcriptomes, proteomes, and multivariate phenotypes. At the same time, society's need for the results of biological research has never been greater. Solutions to many of the world's most pressing problems—feeding a global population, coping with climate change, preserving ecosystems and biodiversity, curing and preventing genetically based diseases—will rely heavily on biologists, collaborating across disciplines.Theodosius Dobzhansky famously proclaimed that “nothing makes sense in biology except in the light of evolution." Though Dobzhansky's statement is sometimes dismissed by biologists in other fields as self-promotion, recent advances in many areas of biology have shown it to be prophetic. For example, genomics, which emerged mostly from molecular biology, is now steeped in evolutionary biology. Evolutionary theory helps to explain our origins, our history, and how we function as organisms and interact with other life forms, all of which are crucial to understanding our future e.g., . Evolutist CenturyA New Biology for the 21The pervasive relevance of evolution can be seen in the 2009 report commissioned by the National Research Council of the National Academies, It's hard to overstate evolutionary biology's power to explain the living world and our place in it. Many applications of evolutionary theory and methods—from animal and plant breeding to vaccine development to management of biological reserves and endangered species—affect society and promote human well-being Our ability to apply evolutionary concepts to a wide range of problems has never been greater. Changes in the availability of data and an emerging scientific culture that embraces rapid, open access to many kinds of data , along with a computational infrastructure that can connect these rich sources of data , will trPeriodically, groups of scientists meet to identify new opportunities in evolutionary biology and associated disciplines e.g., . Rather The new field of “evolutionary medicine" Feeding the rapidly growing human population, especially with increasing stress on agricultural systems from climate change, continues to be a major challenge. The green revolution, from the 1950s onwards, rested on selective plant breeding for larger yields and was underpinned by evolutionary theory Evolutionary approaches have often been applied to the conservation of species and ecosystems Models of mutation, inheritance, and selection have inspired the development of computational evolutionary algorithms that are used to solve complex problems in many fields Genealogical relationships bear on many court cases. Is the defendant really the parent of the plaintiff? Does the evidence  at the crime scene tend to exonerate or implicate a suspect? Evolutionary methods, particularly population genetics, are now used frequently in forensics and court cases to test the link of crime scene evidence to individuals Divining the direction of future scientific research is always fraught with difficulty. Nonetheless, we feel that it is possible to identify some general themes that will be important in coming years. We also present some examples of classic research problems that remain unsolved and that might well be the focus of future work, as well as new and important questions for which evolutionary approaches may be key.The flood of data in all areas of evolutionary biology poses important theoretical challenges: new kinds of theory are sometimes required to make sense of new kinds of data. We can already point to certain broad areas of evolutionary biology that will likely demand sustained theoretical work. These include the elaboration of more formal theories for evolutionary developmental biology ; the more complete incorporation of the roles of epigenetics, behavior, and plasticity in models of trait evolution; analysis of units of selection; and attempts to construct a quantitative and predictive theory that describes the genetic basis of adaptation. In other areas, problems will likely be more statistical than theoretical. Indeed, the enormous quantity of genome data poses serious statistical challenges even for fields that already possess strong theoretical foundations, such as evolutionary genetics.DNA sequencing can now generate whole-genome data not only for single representatives of a few species but for multiple individuals from multiple conspecific populations and even from entire communities. Such multilevel data are giving rise to whole new fields of study  as well as to new theoretical, computational, and data management challenges.C. elegans and yeast One particularly exciting avenue of research afforded by new genomic technology is the possibility of directly observing the dynamics of evolution. In the last few years, genomic analyses of experimental evolution have yielded new understanding of how RNA molecules, viruses, and bacteria evolve . Precise information on place, time, and reproductive stage can be integrated with data on local environmental conditions, often obtained from remote sensing While the explosion of data on genome sequences has received the most attention, supplementing these data with information on the natural history of individuals, species, and their environments will be important. Core information from field-collected specimens always includes species identity and place and time of collection, but increasingly, this information is being enriched with links to field notes and phenotypic , behavioral , and genomic data in a variety of databases  phenotypic and not merely (discrete) DNA sequence data—as well as on integration across databases to allow synthesis of very different kinds of data see . The culAs noted at the outset, we live in an exciting time for evolutionary biology. The field has embraced the “omic" revolution, and answers to many classic questions, which have motivated research for a century, are now within reach. The study of evolution, which in the past was often equated with changes in gene frequencies in populations, has become more holistic and integrative. Researchers are increasingly interested in exploring how interactions among genes, individuals, and environments have shaped the evolutionary process, both at micro- and macrolevels. At the same time, large challenges such as global warming, novel infectious diseases, and threats to biodiversity are increasing, and the opportunity for evolutionary biologists to contribute to their resolution has never been greater.Realizing the full potential inherent in evolutionary biology is, however, far from assured. The task of integrating evolutionary knowledge within and across scales of biological organization, as discussed above, requires development of many comparative databases and analytical tools. We would do well to collaborate broadly, cultivating new expertise, and to watch out for the unexpected, as analyses of new kinds of data can reveal that preconceptions are unfounded.Because most of our science is supported by limited public funds, evolutionary biologists and ecologists should support and participate in efforts to help the public understand the issues and the value of scientific understanding. Science in general and evolutionary science in particular are often politicized, exactly because of their fundamental importance to human society. The next 20 years hold the promise of a golden age for evolutionary biology. Whether we realize that promise depends in part on how effectively we communicate that message.Cyberinfrastructure—The research environments that support advanced data acquisition, data storage, data management, data integration, data mining, data visualization, and other computing and information processing services distributed over the Internet beyond the scope of a single institution. In scientific usage, cyberinfrastructure is a technological solution to the problem of efficiently connecting laboratories, data, computers, and people.Evolutionary developmental biology—The study of the evolution of development, often by the comparative study of gene expression patterns through the course of development in different species.Evolutionary genetics—Population and quantitative genetics.Gene network—A flow diagram describing the interactions among genes during development that affect a particular phenotype or set of phenotypes.Genomics—The study of the entire complement of DNA in organisms (Genome), including is sequence and organization.GMO—Genetically modified organisms in which the genome has been deliberately changed; transgenic organisms resulting from DNA manipulations.Lateral  gene transfer—Genetic transfer between species, as opposed to vertical gene transmission from parents to offspring in a lineage.Metadata—Data associated with individual DNA sequences or organismal specimens .Model organism—Organisms whose genome has been sequenced and for which sophisticated tools for genetic manipulation are available.Natural history—The entire description of an organism, including its phenotype, genome, and ecological context .Nonmodel organism—Organisms whose genome has not been sequenced and/or for which sophisticated tools for genetic manipulation are not available.Ontology—The naming of categories, especially of the functions of genes.Population genetics—The study of the evolutionary forces that change the genetic composition of a population; the discipline is often concerned with evolution at one or a few genetic loci.Quantitative genetics—The study of the inheritance and evolution of traits that are typically affected by many genetic loci.Transgenic tools—Tools that enable the deliberate transfer of DNA sequences from one organism to another or the deletion or modification of DNA sequences, in every cell, in one organism.Figure S1An example of the enormous phylogenetic trees that soon will represent the norm in phylogenetic analyses. This is the consensus tree of the maximum likelihood phylogenies for 55,473 species of seed plants with the location of significant shifts in species diversification rates marked in red across the tree. Adapted from (TIF)Click here for additional data file.Figure S2The Phenomobile, a remote sensing field buggy, and the Blimp, for remotely imaging an entire field. The Phenomobile integrates a variety of remote sensing technologies for measuring phenotypic variables on many plants simultaneously. The buggy straddles a plot and collects measurements of plant temperature, stress, chemistry, color, size and shape, as well as measures of senescence. The Blimp is designed to image all the plants in an entire field from a height of 30–80 m using both infrared and digital color cameras. These technologies were developed by David Deery of the High Resolution Plant Phenomics Centre at the Commonwealth Scientific and Industrial Research Organisation in Australia. Photo credit: Carl Davies, CSIRO Plant Industry.(TIF)Click here for additional data file.Text S1Training to sustain evolutionary biology.(DOCX)Click here for additional data file.Text S2Infrastructure needs and opportunities in evolutionary biology.(DOCX)Click here for additional data file.


The safety of drinking water is evaluated by the results obtained from faecal indicators during the stipulated controls fixed by the legislation. However, drinking-water related illness outbreaks are still occurring worldwide. The failures that lead to these outbreaks are relatively common and typically involve preceding heavy rain and inadequate disinfection processes. The role that classical faecal indicators have played in the protection of public health is reviewed and the turning points expected for the future explored. The legislation for protecting the quality of drinking water in Europe is under revision, and the planned modifications include an update of current indicators and methods as well as the introduction of Water Safety Plans (WSPs), in line with WHO recommendations. The principles of the WSP approach and the advances signified by the introduction of these preventive measures in the future improvement of dinking water quality are presented. The expected impact that climate change will have in the quality of drinking water is also critically evaluated. Water Safety Plan Manual . Within an EU research financed project  one of the objectives was to train water companies on the principles of the WSPs, the experience demonstrated that the companies acknowledged the benefit of the new approach, and some of the largest water companies have incorporated it already. However, small water companies would only be implementing WSPs when the approach becomes a mandatory requirement under the new EU legislation. In reality legislations are good driving forces for such improvements as outlined previously. For instance the national Spanish legislation on Legionella promulgated after the world largest outbreak, requires a part of the microbiological controls two other obligations: (i) to pass a training course for those responsible of handling installations at risk of propagating the legionellosis; and (ii) to implant control plans based on the methodology of HACCP. According to Bartram et al.  in which specific case studies are presented. A dedicated web site on the WSPs has been developed by the International Water Association (IWA) . In addition, WHO and IWA developed guiding documents to initiate such process considering all levels of resources available, so that it can be implemented all over the world, even at poorly developed countries: .Further information, of how to implement a WSP, can be found in the WHO technical guidance documents  ,74,127. et al.  have stuet al. . Floods et al. . Even thet al. ,129. In et al. .http://www.who.int/water_sanitation_health/publications/9789241598422_cdrom/en/.Fishes, green algae and diatoms are often used as water quality indicators of climate changes in waters. Daufresne and Böet  observedi.e., diarrhoea) auto limits by itself without treatment in most healthy people. However, the older and immunocompromized people are at higher risk. Today we also know that this type of infection may have important sequels . The microbiological controls applied to drinking water, have relied on the analysis of faecal pollution indicators in the finished dinking water. The classical indicators have served together with the improvements on the treatment and disinfection to control waterborne outbreaks. However, the use of these indicators may be substituted by the direct detection of pathogenic microorganisms, e.g., in the case of pathogenic viruses [The microbial contamination of drinking water and its control constitutes a major issue worldwide, because it is still a major source of infection and can cause mortality, especially in the children of developed countries, and threatens the health of the population of developed regions, as illustrated by recorded outbreaks. The latter are however, considered to be underestimated because the major symptom ( viruses ,132,133.


Identification of proteins involved in milk production is important to understand the biology of lactation. Many studies have advanced the understanding of mammary function and milk secretion, but the critical molecular mechanisms implicated in milk fat secretion is still incomplete. Milk Fat Globules are secreted from the apical surface of the mammary cells, surrounded by a thin membrane bilayer, the Milk Fat Globule Membrane (MFGM), formed by proteins which have been suggested to be cholesterolemia-lowering factors, inhibitors of cancer cell growth, vitamin binders, bactericidal, suppressors of multiple sclerosis. Using a proteomic approach, we compared MFGM from milk samples of individuals belonging to two different cattle breeds, Chianina and Holstein, representative of selection for milk and meat traits, respectively. We were able to isolate some of the major MFGM proteins in the examined samples and to identify differences between the protein fractions of the two breeds. We detected differences in the amount of proteins linked to mammary gland development and lipid droplets formation, as well as host defence mechanisms. We have shown that proteomics is a suitable, unbiased method for the study of milk fractions proteins and a powerful tool in nutritional genomics. Even when high MASCOT scores were obtained , if proteins were identified by a single peptide a combination of automated database search and manual interpretation of peptide fragmentation spectra was used to validate protein assignments. The mass error, the presence of fragment ion series and the expected prevalence of C-terminus containing ions (Y-type) in the high mass range were all taken into account in the verification procedure. Moreover, protein hit identity was confirmed by replicate measurements.2 µL/min . SequentUsing a proteomic approach, we have successfully identified some of the major MFGM proteins in two different cattle breeds. We utilised a nonbiased method for the discovery of specific molecules involved in this unique secretory function. This work provides a preliminary study of the complexity of the secretion of MFG in mammary epithelial cells during the lactation process. A considerable advantage of the proposed method relies on the possibility of analysing this peculiar kind of cells without the need of biopsy or animal slaughtering, which often represents a limiting factor in sampling, for both ethical and economic reasons. We analysed samples of two cattle breeds selected for different purposes, and our preliminary results indicate that protein differences at the MGF level can be observed even if the animals were raised in the same conditions. In detail, PIGR, having a role in the immunoglobulin function, and GP2, a protein that may serve as a protective against infections, resulted overexpressed in Holstein, which is a highly specialised dairy breed. Dairy breeds are selected, among other traits, for resistance to mastitis and for somatic cell score. This may represent an effect of long time selection and could lead on an insight on the effects of milk of different origin on calf health and on its nutraceutical characteristics. In Chianina beef breed ADRP, a protein involved in lipid deposition, is overexpressed. Further studies should focus on fat amount and composition in the milk of this breed, and on the potential implications of such information for the human diet. Also MFG-E8, a protein involved in the lactation process and showing anti-viral activity, is overexpressed in this breed at the lactation stage analysed. Beyond having antiviral properties, MFG-E8 could be related to the different timing in mammary gland maturation and drying off between the two breeds. Further investigations are needed to better define the difference in MFGM protein composition between Holstein and Chianina breeds and to fully describe the variation of the MFGM proteins during the lactation phases.Our study shows that proteomics is a suitable method for the investigation of the characteristics of milk proteins. Moreover, it is a powerful tool in nutritional genomics, allowing the analysis of milk as food and helping the selection of animals with characteristics relevant for human health.This work was financially supported by the “GENZOOT” research program funded by the Italian Ministry of Agriculture. We wish to thank Stefano Corbianco for providing the samples, Maria Giulia Egidi for help in mass spectrometry analysis and Paolo Ciorba for sampling assistance.


This study examines listener judgments of musical tension for a recording of a Schubert song and its harmonic reduction. Continuous tension ratings collected in an experiment and quantitative descriptions of the piece's musical features, include dynamics, pitch height, harmony, onset frequency, and tempo, were analyzed from two different angles. In the first part of the analysis, the different processing timescales for disparate features contributing to tension were explored through the optimization of a predictive tension model. The results revealed the optimal time windows for harmony were considerably longer (~22 s) than for any other feature (~1–4 s). In the second part of the analysis, tension ratings for the individual verses of the song and its harmonic reduction were examined and compared. The results showed that although the average tension ratings between verses were very similar, differences in how and when participants reported tension changes highlighted performance decisions made in the interpretation of the score, ambiguity in tension implications of the music, and the potential importance of contrast between verses and phrases. Analysis of the tension ratings for the harmonic reduction also provided a new perspective for better understanding how complex musical features inform listener tension judgments. The percept of musical tension provides a window into the disparate components that comprise an expressive performance. Performers' interpretations of a composed piece highlight structural features of the music, shaping the listener's perception of both apparent and unusual aspects of the score Palmer, . In addiTension has long been central topic of interest in music theory , which combines elements of Narmour's , 1992 imIn addition to Huron's work, there have been other researchers who have suggested links between tension and affective arousal , make it possible to investigate these responses in greater temporal detail than has previously been statistically defensible, providing new insights into the time course of tension ratings and agreement between responses.In this paper, we focus on two previously unexplored aspects of musical tension: (1) the individual timescales at which disparate parameters contributing to tension are processed, and (2) detailed differences in how participants rate similar excerpts. We examine tension from two very different angles—from the perspective of the average tension response and from the ratings identifying tension-related differences between the stimuli. On both accounts, novel approaches to analyzing continuous tension data are explored, providing new insight into how tension is perceived and processed. From a broader cognitive point of view, timescales for feature processing can be viewed from a “levels-of-processing” perspective , 11 female, 14 male. Subjects had an average of 9.78 years of formal training on a primary musical instrument (SD = 5.18) and an average self-rank in instrumental skill level of 3.90 (SD = 0.94) on a scale of 1–5. Average number of semesters of college-level music theory training was 3.32 (SD = 2.43) and the mean overall self-ranked musical training level was 3.52 (SD = 0.92), where 0 = no training and 5 = professional-level training. The four outlier participants were excluded for not completing the tension-rating task. In the first case, the participant became bored and started hitting random keys, resulting in the data collection interface exiting prematurely; in the other cases, the subjects were unresponsive and did not indicate any tension changes during some or all presented stimuli.A total of 29 subjects took part in the experiment, of which four were excluded from this analysis. The remaining 25 participants were primarily undergraduate and graduate students at New York University, mean age 25.16 years . These other stimuli were an original 4′15″ excerpt from a Brahms piano concerto and three scrambled versions of it. The two stimuli used for the current study are a recording of “Morgengruss” from Schubert's Participants were seated in front of a computer and presented the stimuli over Sennheiser HD 650 headphones in a hemi-anechoic chamber. They were asked to indicate changes in musical tension while listening to the stimuli by moving a horizontal slider on a MATLAB GUI that used Psychtoolbox extensions .p-value from goodness-of-fit tests on multiple variants of the time series segmented into frames, with data-driven algorithmic controls on the independent model and distribution testing . The coordination test is evaluated using non-overlapping time frames of size greater than the sampling interval. To reduce the impact of arbitrarily cutting up the time series in even time frames and the risk of splitting apart changes related to the same event, the coordination score is calculated from all distinct slicings of the time series into adjacent non-overlapping time frames of fixed duration. The details of the calculation are accessible via the activity analysis MATLAB toolbox . Very high coordination scores indicate that across the responses, activity is very well coordinated and strongly suggests repeatable summary time series, however, this does not mean that all ratings are in complete agreement on how and when tension changes.Built on this process of evaluating coordination of activity in a collection of time series is the Coordination Score. It is helpful to consider which collections of responses, e.g., responses to different stimuli, show more or less coordinated activity. The coordination score is calculated from the averaged negative log of the x Upham, . The cooThe principle of the coordination score for single-activity events can also be applied to evaluate the independence of different forms of activity in the same collection of responses, for example, both increases and decreases in tension ratings . If a collection is coordinated in their activity, increases and decreases should alternate, but if the responses are independent, both types of activity are likely to happen in the same time frames. The coordination between two types of activity are similarly quantified and scored. If a collection of ratings is coordinated in each direction of tension change but fails to reject independence of increases and decreases, this undermines the robustness of the average time series. Its temporal profile, most often the object of analysis in relating continuous-response data to the time course of the stimulating music, is not likely to be robust since contributions of individual responses cancel out rather than strengthen the common progress of tension.In the analyses below, activity-level time series are used with average time series to describe the four collections of tension ratings, while the coordination scores, rating change increases, decreases, and alternation between the two, provide arguments for which response collections are employed for modeling and comparisons between verses.As a first step in the analysis procedure, seven musical and expressive features were quantified for the Morgengruss performance: dynamics, tempo, onset frequency, harmonic tension, and pitch height of the melody, inner voice, and bass line. Although this is not an exhaustive list of features, it was deemed sufficient to account for most of the variation in the tension responses.The note onsets of the vocal line and accompaniment were determined using marker references in Amadeus Pro  in conjunction with the audio-editing program Audacity (V 2.0.0) for locating precise onset times. With some musical discretion, onsets were generally marked at vowel onsets in consonant lead syllables and piano onsets were also included whenever they did not coincide with the vocal line. Onset frequency was calculated directly from the onset times and was quantified as the difference between the maximum event duration in the performance (3.51 s) and the current note-event duration. Thus the shorter the length of the note event was, the higher the onset frequency. Tempo was determined by using beat onsets. In cases where there wasn't an onset available on a beat, the onset time was linearly interpolated from the positions of the last onset and the next available onset.Pitch height of the melody, inner voice, and bass lines were initially encoded as MIDI values and aligned to onset times. The inner voice is only active in the third phrase where the accompaniment echoes the vocal line; however, it was deemed prominent enough to merit addition to the feature set. A harmonic tension graph was created by using the mean tension response to the harmonic reduction stimulus . Tension values sampled 2 s after the onset of each chord were used to quantify harmonic tension for the entire piece . For pitch height, the mean and standard deviation for all three melodic lines combined were used to normalize the graphs. The normalized mean tension response and features are shown in Figure The mean tension response and feature graphs for loudness, tempo, onset frequency, and harmony were then normalized to zero-mean and unit standard deviation  a memory window, which immediately precedes the attentional window and represents an abstraction of a previous tension trend; (3) differing weights for each of the musical parameters.The first analysis approach explored the timescales at which performers and listeners process individual musical features and structures. The vehicle for this analysis was a modified version of Farbood's  trend satrend salience of musical features is the key determiner of tension judgments. The model predicts tension as a function of individual musical features by taking into account the combined directional change of all of the features in the attentional window. This combined directional change, or tension trend, is weighted by what immediately precedes it in the memory window—if the direction of the trend in the memory window matches the current attentional window trend, the magnitude of the cumulative tension trend is additionally increased. These trends are integrated over time to generate a final tension prediction.The concept of tension trends is the theoretical core of the model, describing how individual musical and auditory features integrate and interact to produce a global feeling of changing tension. The idea is that the t is defined assf(t) is the slope of best linear fit of feature f at time t; β = 1 if the sign of s(t − d) does not equal the sign of s(t); β is some positive value, empirically determined, if the sign of s(t − d), where d represents the duration of the memory window, is the same as the sign of s(t); wf is the weight of feature f with ∑fwf = 1. See Farbood . Furthermore, the feature weights were not used—differences in feature contributions were solely determined by the individual timescales.This model was modified for the current study in one significant respect: instead of employing fixed attentional and memory windows, different time windows were used for each feature in order to optimize the model predictions. Farbood's  evaluatiTo translate between the recorded stimuli and metrical time, timings for every 1/36th of a measure were interpolated from the onset times in the performance, accommodating quadruple and triple subdivisions of quarter notes. Linear interpolation had to be used in absence of a better model for perceived tempo between onsets, since the resulting inaccuracies would be too small for the timescales employed in the analyses below. This mapping translated participants' tension ratings and musical features, initially sampled at 10 Hz, to metrical time with nearest-neighbor interpolation. Counting measures from the beginning of the verse, the harmonic reduction omitted mm. 7, 18, and 19, since they were repetitions of preceding chords. Matching these ratings to the verses, gaps were either interpolated or left blank, depending on the analysis.p value for that time frame, thus providing a local estimate of likelihood for the experimental activity at each moment. Those moments with activity levels exceeding 95% of the alternatives have been marked as having significant coordination. By this method, it is possible to assess overlapping time frames (non-overlapping time frames are used in the collection coordination score). The basis of this approach is to assume that rating changes are not related to the stimulus—that they are noisy signals rather than clean—and that the points selected are those that have strong evidence to the contrary. This conservative perspective is necessary so long as we do not have a model for forecasting activity levels and the noise in rating changes with more precision. For now, moments of high activity coordination can be considered likely driven by the common stimulus, but moments of less notable activity levels should not be dismissed as noisy and unrelated to the music.While global measures of coordination are useful for confirming shared behavior between ratings of tension to the same stimulus, they are insufficient for comparing ratings to related stimuli with any temporal sensitivity. For that, we employed a nonparametric method for generating distributions of activity levels for each time frame against which actual activity level could be assessed. First, each response time series was reduced to a point process, a sequence of 0 s and 1 s, where 1 indicates the activity in question, such as an increase in tension rating from one sample to the next. Shift values for each response were randomly selected from a time interval , yielding an alternate alignment of these point processes by shifting the complete series in time by the corresponding time value (with the ends looped for continuity), and the activity levels for each time frame were calculated as for the original alignment  and was integrated in 1 s attentional window frames. The goal was to optimize the output of the model by adjusting the window durations for each feature and correlating the prediction results with the empirical data.The model was trained on the first half of the data (consisting of the piano introduction and first two verses) and then tested on the final two verses. From a theoretical perspective, it was assumed that the timescales for the three pitch-height descriptions should be identical. That resulted in five memory and attentional window durations to optimize, totaling 10 variables, each with 42 possible durations (0–20 s in 500 ms increments). This high-dimensional space is too large for an exhaustive search, so a step-wise testing procedure was implemented to explore the state space. Starting with all variables at the minimum duration of 0 s, one feature at a time was incremented until the correlation between the model output and mean tension response reached a peak. After optimal values were found for all features, the model output was tested again by exploratory deviations from the fixed optimal values in order to provide further confirmation of the result.Given this heuristic approach, it is possible that there exists a better solution than what was found—i.e., that the resulting values represent a local maximum. However, the contributions of each feature appear to be predictable around individual maxima for each variable, indicating that a better solution is unlikely. The optimal values found, listed in Table These values, optimized only for the training data, were then used to produce a tension prediction for the test data. This also resulted in a strong correlation with the mean tension response, ρ = 0.79, providing some evidence that the model was not overfitting the data. This comes with the caveat that it cannot be determined with certainty that overfitting did not occur since the training and test data are very similar both in terms of the tension responses and feature descriptions . See Figure The next part of the analysis entailed an in-depth comparison of tension ratings between different presentations of related musical material, namely the individual verses and the harmonic reduction. Given the differences between the stimulus excerpts, we first considered statistics summarizing the cohesion of the collections of tension ratings per excerpt in their second presentations, including rating-change activity coordination. Second, we explored the summary times series for these tension ratings, as aligned to the common musical time course, examining differences between mean tension ratings and moments of extreme coordination. These analyses were primarily descriptive and speculative, demonstrating new approaches to explore and compare continuous ratings of tension. The results provided insight into continuous ratings of tension, potential features to explore in modeling efforts, and interesting examples of musical moments with counterintuitive rating-change behavior.As stimuli, the harmonic reduction and the performed piece are substantially different in character. The harmonic reduction is limited in timbre, with a steady tempo, uniform loudness of notes, very sparse onsets of notes with little impression of meter. Its relationship to the performed verse is restricted to the harmonic sequence, the piano as a sound source, and the approximate duration. As such, it is remarkable how similar the tension ratings were to those of the performed piece, at least as captured by the average.The verses differ in lyrics, expression, structural order, and other performance parameters. As the participants in the study were US residents, we presume that most if not all were unable to understand the German text. The impact of the lyrics on the differences in tension ratings between verses were therefore likely related to the non-verbal aspects of the musicians' performance. The analysis of tension-rating differences between verses were thus focused on features that were accessible to listeners regardless of language comprehension: articulation, tempo deviation, timbre, and other aspects of affective expression.Ignoring the temporal sequence for the moment, Table Looking at the aligned average tension ratings in Figure Across verses, the loudness and tempo of the first phrase (mm. 1–7) were similar or greater than those of the second phrase, mm. 8–11. Despite this, the second phrase always had a higher tension range than the first. The harmonic reduction offers one possible explanation for this pattern, as the tonal character of the second phrase is farther removed from the tonic. Although loudness and tempo have often been noted to have significant influence over the perceived tension and intensity of music, this moment demonstrates that other features can contribute independently.With regard to absolute tension-rating values, a few peculiarities of the ratings for the harmonic reduction require some explanation. As mentioned before, these values included the beginning of the rating task, the period when participants moved from the initial start position to a stimulus-related value. The relatively steep rise over mm. 1–3, shown in Figure p < 0.05), as described in section Local Coordination Analysis. The following discussion will concentrate on these “significant” moments as a way to contrast ratings for each verse and the harmonic reduction.Additional interesting contrasts can be seen in the rating-change activity levels in Figure Rating-change activity levels for the harmonic reduction are shown in black in Figure The last verse of the performance had the highest average tension values of all excerpts, as reported in Table Contrasts between repetitions are rarely discussed in models of musical tension, despite their importance in performance practice. Performers generally make contrasts in returning material, particularly when semantic differences such as lyrics support a difference. While the overall tension averages were very similar, there were many other interesting contrasts in rating-change activity between verses that co-occurred with deliberate differences in the performances of the same melodic line and accompaniment.The second phrase (middle graph, Figure The activity levels revealed another layer of complication for interpreting the tension ratings of this second phrase. Although most participants did show a slight increase in tension ratings in mm. 9 and 11, the most concentrated activity occurred in mm. 8 and 10, as can been seen in Figure The specific timing of tension-rating changes to the cadential motion in mm. 16–19 of the performed verses and mm. 16–17 in the harmonic reduction point to the importance of context and clarity of cues. Measures 18–19 in the performed recording were very similar to chords 16–17 in the reduction in terms of harmony, timbre, and voicing. However, the ratings for mm. 18–19 in the performance were never as coherent as those of the harmonic reduction. The ratings for the performed verses anticipated the final event, a tonic arrival, decreasing considerably before its onset. There are two plausible explanations for this: metrical anticipation and contextual richness. As mentioned above, the time between events in the harmonic reduction was too long for participants to predict the exact moment of the next downbeat. This lack of definite pulse would have forced participants to wait for cues, instead of anticipating them. In contrast, the performed version provided listeners with a rich metrical framework. Prepared by temporal expectations, they may have been reporting decreases early in anticipation of a downbeat that had been displaced by the expressive rubato of the performers. Alternately, these decreases may have been the result the stronger cadence in mm. 16–17 preceding this moment, during which participants also indicated tension decreases before the arrival of tonic. Other cues present in mm. 16–17 in the performed version could have encouraged tension-rating decreases before tonic arrival, including dynamic cues in the vocal line, which include timbre and loudness tapering over the last sung syllable of the verse. Like the confusion in m. 10, these moments are reminders that much of perceptual effects of music are not easily deduced from the notated page.There are many other nuances of performance that could be explored using activity-level time series since they allow for the identification of salient moments of rating change. With more precise representations of tempo variation, articulation, and vocal timbre, it may be possible to quantify the factors behind inter-verse differences in m. 2, the transition to m. 4, the transition to m. 6, and at the end of m. 8. The timing of changes in this collection of responses point to the relevance of context—structural, metrical, and harmonic contrasts—between successive verses and phrases.This study examined listener judgments of musical tension for a recording of the Schubert song Morgengruss and its harmonic reduction. We focused on two previously unexamined aspects of tension: the differences in processing timescales of disparate expressive and structural features contributing to tension and differences in tension ratings between the verses and the harmonic reduction. The methodologies employed in both set of analyses were novel, providing a fresh perspective on how tension reflects listeners' and performers' musical perceptions.The first part of the analysis examined timescales of musical feature processing by using a modified version of Farbood's  trend saThe results indicated that harmony was processed across a far longer time span than all other features, having a combined attentional plus memory window duration of 20.5 s. The feature with the next-longest combined window duration was pitch height at 4 s. Onset frequency, tempo, and dynamics all had combined window durations of ~2 s, however, dynamics had an optimal memory window of 0 s, indicating that loudness induces the most instantaneous tension response. These results align with more general perspectives on working memory such as Craik and Lockhart , who theIt is perhaps no surprise that listeners are responsive to highly local changes in loudness. Instinctual response to loudness is a basic, low-level function—anticipating and sensing approaching and retreating objects in the environment is important from an evolutionary perspective; in particular, listeners are highly sensitive to looming sounds Neuhoff, . Tempo pThe optimal window durations for pitch height were approximately twice as long as those for tempo and onset frequency. This reflects the likelihood that melodic contour is processed at a higher level of abstraction than all the other features examined here except harmony. Gestalt perception is a primary factor in melodic contour perception (cf. Meyer, The process of tonal induction and harmony perception requires higher-level cognitive abstractions than any other features, and this is evidenced by the long optimal time windows. These findings are in concordance with the result of a preliminary study by Farbood  that exaThe second part of this study examined the differences in tension ratings for the varying interpretations of the Morgengruss score across the four performed verses and the harmonic reduction. While the average tension ratings for the verses were very similar, differences in how participants reported tension changes support the importance of performance decisions that provide nuance to the interpretation of the score. Performer-controlled features such as loudness and tempo, less-easily-quantified articulation and sustain, and contrasts between successive verses were each highlighted as likely factors for these subtle but substantial differences in the tension ratings.Analysis of ratings of the reduction underscored the importance of harmony by providing an explanation for the high tension ratings in the second phrase of each verse. It also offered an interesting case study for analyzing activity in tension ratings; having a reduction representing at least one complex feature made it easier to make sense of the complicated information contributing to the dynamics of average tension ratings.There is some precedent for comparing tension or other continuous ratings between interpretations of a common work (Fredrickson and Johnson, While the coordination analyses of the tension-rating changes were data-driven and robust, the comparisons between responses across different stimuli were not performed systematically. Interpretation of contrasts between verses were informed by musical experience and need to be tested, perhaps with other examples of controlled stimuli. The study of Romantic lieder in particular would be greatly improved with the incorporation of a dynamic model of tempo prediction, sensitive to the grouping implications of different degrees of rubato and descriptors of the sung timbre and articulation.Comparisons of the tension ratings between verses also raises the question of structure and tension judgments, in particular, the importance of contrast between verses. How the same moment in a verse is treated from one iteration to the next warrants more attention since it might be a means to capture the role of form in musical memory and the continuous experience of music. What would it mean for a model to include information about the previous presentation of the same material? While the idea of contrast between repetitions affecting tension is intuitive and familiar to performers, testing this hypothesis would require more instances of specially composed stimuli.The effectiveness of any analysis of tension is dependent on the inclusion of a complete set of features contributing to tension. Although we examined several key parameters, they do not represent an exhaustive set of features that account for all tension variations perceived by the performers and listeners of the Schubert. Perhaps the most significant feature that requires future examination is timbre. Defining the perceptual dimensions of timbre is a difficult task, and that is perhaps one reason why timbre is underexplored in the tension literature. Prior studies that have explored timbral tension in some capacity have looked at features such as roughness, brightness, spectral flatness, and density (von Helmholtz, Although additional experiments using more varied musical stimuli would undoubtedly strengthen and solidify these findings, the analyses and observations made in this study present new perspectives on tension. They help illuminate the complex process of how tension is affected by performance decisions and how listeners respond to those differences. Having a deeper understanding of tension perception can help us better grasp the interplay between expressive performance, listener interpretation, and musical structure.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Overtrading is a common anomaly among stock investors. This study examines the relationship between overtrading and investment returns and the impact of the Big Five traits and gender on overtrading in a unilateral trend stock market using a simulated stock investment system. The data were derived from a sample of undergraduates from six universities who performed in a simulated stock investment situation and had their personality traits measured by the Big Five Personality Questionnaire. The results indicate that: (1) Overtrading was significant in rising stock markets, but not significant in falling markets. (2) The degree of female investors who overtraded was significant in rising markets. (3) The degree of overtrading investors who were high in extroversion or agreeableness was significant in rising markets. The implications of these results for more effective investment strategies are discussed. Overtrading is a common anomaly among stock investors, as observed in behavioral science research. As early as 1968, Jensen found that the returns of the most actively traded mutual funds were lower than the market rate of return Do stock investors trade excessively? Does large trading volume reduce investment returns? Do market situations, personality and gender affect trading volume and thereby affect returns? Early studies were unable to reach an agreement on these questions. Insight into these issues would guide investors to adjust their investment behavior and increase their investment returns.Studies investigating securities investment decisions are difficult to perform because an investment decision is similar to a ‘black-box’ in which the identity, asset scale, investing activity and return from a trade are sufficiently ambiguous that needed information is difficult to access. A number of researchers are able to obtain data from brokerage companies, but most of the information is dated. In the real market, the behavior of investors is affected by many factors, such as experience, knowledge and the market situation. These uncontrollable factors render research results unreliable. This article uses the simulated stock trading system developed by the University of Science and Technology Beijing to gain instant and accurate trading data. In the simulated system, the participants are allocated a certain amount of fictitious capital and stock that they can use to trade in a virtual network, thus affecting the price trend of the stocks. As the researchers can hide behind the network and act as a large institution to manipulate increases and decreases of stock prices, the real-world stock market can be simulated more closely. This type of simulated system is, by itself, an innovation in the study of experimental economics because it can accurately reflect the choices of participants when they make investment decisions. More importantly, the system allows variables to be manipulated experimentally and enables experimental conditions to be controlled, thereby ruling out unrelated factors and duplicate tests while simultaneously overcoming certain limitations of empirical studies.In a conclusion that summarized the micro-level foundations of behavioral finance, DeBondt and Thaler Drawing lessons from the above research, overtrading is defined in this paper by a negative relationship between trading volume and investment returns. When returns are significantly negatively influenced by trading volume, overtrading occurs; otherwise, overtrading is not present.Is trading hazardous to one’s wealth? Our study addresses this issue by examining the relationship between trading volume and investment returns, as well as the influence of the investment situation, the Big Five personality dimensions and gender on overtrading. In this article, different investment situations were developed by manipulating stock prices, and the Big Five personality dimensions of participants were measured.In reality, the trading behavior of stock market investors is influenced by rising and falling markets. Shefrin and Statman These findings indicate that stock investors make different decisions when facing different investment situations, as measured by gains and losses. Specifically, when making decisions in possible gain situations, investors are risk-averse, selling large amounts of stocks when prices go up and becoming likely to trade excessively. In contrast, when making decisions in possible loss situations, investors tend to be risk-seeking and therefore do nothing when prices go down. The first hypothesis of this paper is the following:Previous studies explored the relationship between personality traits and investment behaviors using an experimental economics approach. Chinese scholars Peng & Wang Extroversion reflects the involvement of individuals in the outside world. Individuals with high extroversion scores are energetic and sociable, while individuals with low scores are independent and introverted Agreeableness refers to individuals’ courteousness, trustworthiness, tolerance, compassion, generosity and cooperative demeanor Conscientiousness reflects an individual’s tendency to act in an organized, effective, reliable and self-disciplined manner Neuroticism reflects an individual’s tendency to experience negative emotions; high-N individuals are apt to irrationally overreact to bad feelings and therefore experience psychological effects such as anxiety, pressure, hostility, tension, anger and depression Openness is a personality dimension that represents the difference between conservative thought and open thought. It is the individual’s tendency to pursue novel, artistic, flexible and intellectual factors Previous research has noted that gender influences investors’ trading behavior; specifically, men behave over-confidently, while women do not Experiments in this paper took place in the financial engineering laboratory of Dongling School of Economics and Management at the University of Science & Technology Beijing. This platform provided technical support for both hardware and software. The hardware included the servers and all the computers needed for the participants, while the software included the simulated stock trading system developed independently by the laboratory.One server was used as the web interface server, and another server was used as the dealmaker server for the simulated stock trading system. Using the web interface server, participants in this experiment could determine their asset balance, make deals, delegate and revoke deals. The trading information then flowed into the dealmaker server, where buy and sell deals were matched according to the principle of price priority and time priority, and unmatched trading could be revoked by the participants.In this research, the trading system used was a “T+0″(The settlement date of security transactions and denote that the settlement and delivery takes place immediately after the transaction occurs. To settle means to exchange the shares (stocks) for the cash and vice versa.) delivery and settlement system. Due to the experimental time limit and to set free the volatility of the stock prices and record the returns of investors with different risk attitudes, there were no tightening constraints on the price-changing margin, such as 5% or 10%. In this research, the limit of the price-rising margin was 500%, while the limit of the price-falling margin was 99.9%.Unilateral movement is a common and simple trend in the stock market that indicates either a unilateral rising or a unilateral falling in stock prices. This research is based on this simple stock trend to analyze the impact of investment situations, Big Five personality dimensions and gender on overtrading behavior, and leaves the analysis of more complicated stock trends to further studies.The experimental platform only allowed 160 subjects to be studied at one time, to ensure that data took on an apparent trend, the research was only designed with one stock. The number of shares issued, shareholders and the financial statements information about the public company can be viewed in the simulation system. Participants can calculate the internal value. Due to the experimental time limit and the main research is on the returns of stock trading, there are no dividends. Each of the participants in each of the experimental pre-openings was assigned ¥200,000 virtual cash and 10,000 shares of stock. To induce some variation across experiments, opening prices were different. After the opening, each test was traded freely at the same time by buyers and sellers; Each investment transaction time was 20 minutes, with a 5-minute break between each field being enacted by shutting down the system.Before the experiment, all the participants were required to complete the Big Five personality assessment scale. Capital and stock were distributed evenly to each participant. The researcher then used a hidden account to manipulate the stock price and force the stock market to be unilaterally rising or falling, leaving the participant investors to make their own investing decisions and the computers to record their positions in trading volume and investment returns.The basic principle for manipulation was to control the stock prices’ stability, or else the unilateral trend would be destroyed. Guided by this principle, some manipulations in these experiments took place in the third minute, while others took place in the fourth minute. Referring to the specific stock price trends, the manipulation in each experiment could form a rough trend because the stock price was also affected by the commissioned price paid by the participants, which was out of our control, as well as the stock trend determined by the equilibrium of price put forward by both buyers and sellers. Consequently, specific stock price trends could be manipulated by the researchers. Therefore, in this research, all stock price trends were treated equally, no matter the manner in which the trends were generated.First, participants were provided with the following instructions: “These experiments will be conducted in a simulated stock investment situation, and the goal of the experiments is to study overtrading and the impact of personality type on the stock trading volume. You will be informed of the results at the end of the course. We designed one stock for trading in the simulation system. The number of shares issued, shareholders and information about the public company available in financial reporting statements can be viewed in the simulation system. Each student has been assigned ¥200,000 in virtual cash and 10,000 shares of stock. You can freely trade stocks after the beginning of the experiment. There are no dividends. The final trading returns will serve as the main reference for the grading of the investment course.” Then, participants conducted 6 experiments under non-identical conditions over the course of two weeks. The first three experiments were conducted in a unilaterally price-rising situation, while after two weeks, the following three experiments were conducted in a unilaterally price-falling situation.To avoid the influence of the experimental market experience on the behavior of investors, the same participants conducted two further experiments with an inverted treatment order . Experimental platforms and parameters were the same as the previous experiment, with the only change occurring in the order of experimental situations. The first experiment was conducted in a unilaterally price-falling situation while the following was conducted in a unilaterally price-rising situation.The 115 participants in this experiment were students in the selected investment course at the University of Science and Technology Beijing; the students came from different specialized subjects from 16 universities. Among the participants, 43.5% were male and 56.5% were female. The students were permitted to communicate with one another and to trade online at the same time. Each experiment lasted 20 minutes, and there was a 5-minute break between each experiment. We employed standard psychological methods to study the relationship between personality and returns, which would not cause physical or psychological damage. We followed the guide of the Chinese Ethics Committee of Registering Clinical Trials and received the approval of Student Affairs, which was responsible for the safety of students and was a formal organization at the University of Science and Technology Beijing. Due to the number of students and because the students were online during the experiments, we only received the students’ oral consent; however, we also received the approval of the teacher in charge, and the students were informed that the purpose of this experiment was to study overtrading and the impact of personality and gender on trading volume before the experiment was begun, After the experiment, the results were shared with the students.We used the five broad Big-Five personality domains (50 items) from International Personality Item Pool (IPIP) In this paper, trading volume was denoted as the number of stocks being traded by individual investors. The sum of the trading volume in the three experiments with a unilaterally rising stock price was treated as the trading volume in the investment situation with a unilateral increase in stock price. Similarly, the sum of the trading volume in the three situations with a unilaterally falling stock price was treated as the trading volume in the investment situation with a unilateral decrease in stock price. Meanwhile, the average return in each situation was treated as the return in the unilaterally price rising situation and the unilaterally price falling situation, respectively. The related formulas are:TV in UPR = sum of TVs in three corresponding UPR experiments.TV in UPF = sum of TV in three corresponding UPF experiments.R in each experiment = (ending assets − beginning assets)/beginning assets.R in UPR = average R in three corresponding UPR experiments.R in DPI = average R in three corresponding UPF experiments.TV stands for Trading Volume and R stands for Returns, while UPR stands for a Unilaterally Price Rising situation and UPF stands for a Unilaterally Price Falling situation.The software used in the statistical analysis was SPSS19.0.The stock price trends generated in the 6 experiments are shown below:The two experiments with an inverted treatment order  are shown below:Descriptive statistics and the results of correlation analysis of returns and trading volume are presented in Descriptive statistics and the results of correlation analysis of returns and trading volume with respect to the inverted order treatment are presented in Descriptive statistics and correlation analysis of personality and gender on trading volume are shown in The trading volumes of stock investors are always high, especially in the Chinese stock market. Because of these high trading volumes, many researchers assume that there is overtrading in the stock market. In this paper, we collected data from a simulated stock trading system and performed a statistical analysis on these data, finding that in unilaterally price-rising situations, there is a negative relationship between returns and trading volume. In other words, there was overtrading in the stock market. However, in unilaterally price-falling situations, there was no significant relationship between stock returns and trading volume, and overtrading did not occur. This result is consistent with Hypothesis 1, which stated that the investment situation would influence investing behavior, thus supporting the “disposition effect” and the “prospect theory” proposed by Shefrin and Statman The data analysis in this paper also indicates that investors’ trading volume is not influenced by the Big Five personality dimensions of investors. However, in unilaterally price-rising situations, the trading volume of an investor with high scores in extroversion and agreeableness could adversely predict stock returns. These results are consistent with Hypotheses 3, 4 and 6 but not Hypotheses 2 and 5. In Hypothesis 2, we hypothesized that extroverted investors may be more rational and therefore less prone to overtrading. However, the results show that investors with high extroversion scores trade excessively in unilaterally price-rising situations. Li and Liu Regarding the analysis on gender’s impact on trading volume, we found that the trading volumes of male investors are higher than those of female investors in both unilaterally price-rising and price-falling situations. This result is consistent with previous research indicating that male investors are more confident than female investors and trade more than female investors do by up to 45% This paper focuses on the impact of the investment situation, personality and gender on overtrading. Due to the complexity of personality and the variety of factors affecting investment decisions in stocks, the conclusions from this research should be further investigated. There were some limitations experienced by researchers. First, the investment environment was in the simulated stock trading system. In the real world, there are so many types of stocks and investors that stock prices are also influenced by investment policy, company performance and the manipulation of institutions. Therefore, in a further study, more stocks should be included in the simulated system to reflect real-world stock markets more closely. Second, there were only 8 experiments that lasted 20 minutes with different manipulation methods, there are limitations to the external validity of the results. To confirm the relationships found in this paper, further research should include more experiments in different situations. Finally, the definition of overtrading in this paper involves the existence of a significantly negative relationship between returns and trading volume, ignoring the effect of market periods in the decision-making process of investors. Therefore, a time-series analysis should be utilized in further studies.Despite these limitations, there are still some reasonable suggestions based on the conclusions in this paper. First, investors should hold appropriate stocks and gain returns in unilaterally price-rising situations because investors tend to sell winners too early and trade excessively in such situations. Second, female investors should decrease their trading volume and avoid early selling in unilaterally price-rising situations because they are likely to trade excessively. Finally, although personality as a whole does not affect trading volume, investors with high scores in extroversion and agreeableness tend to trade too much in unilaterally price-rising situations; thus, these investors should avoid pursuing risk and interference from other investors and decrease their trading volume at such times.Through analysis of the data collected in the simulated stock trading system, we observed that there is a significantly negative relationship between returns and trading volume in unilaterally price-rising situations, while there is no such relationship in unilaterally price-falling situations, which indicates overtrading in the former situation. The trading volume of male investors was significantly larger than that of female investors; however, males did not overtrade in unilaterally price-rising situations, while female investors did. Finally, there was no significant relationship between trading volume and the Big Five personality dimensions, but the returns of high-E investors and high-A investors were adversely affected by their trading volumes, which indicates overtrading by investors with high scores in extroversion and agreeableness.