The interactions between peptides and lipids are of fundamental importance in the functioning of numerous membrane-mediated cellular processes including antimicrobial peptide action, hormone-receptor interactions, drug bioavailability across the blood-brain barrier and viral fusion processes. Moreover, a major goal of modern biotechnology is obtaining new potent pharmaceutical agents whose biological action is dependent on the binding of peptides to lipid-bilayers. Several issues need to be addressed such as secondary structure, orientation, oligomerization and localization inside the membrane. At the same time, the structural effects which the peptides cause on the lipid bilayer are important for the interactions and need to be elucidated. The structural characterization of membrane active peptides in membranes is a harsh experimental challenge. It is in fact accepted that no single experimental technique can give a complete structural picture of the interaction, but rather a combination of different techniques is necessary. Peptide-membrane interactions are involved in numerous crucial biological processes, such as antimicrobial defense mechanisms, viral translocation, membrane fusion, functions of membrane proteins, transport of therapeutic compounds, disruption of integrity of membranes, and others. Membrane interacting peptides comprise a large family of diverse peptides exhibiting a broad range of biological activities and, therefore, continuously attract growing interest for their biomedical applications.During peptide membrane interactions, both the peptide and the membrane may experience a series of structural changes. Thus, theoretical and experimental studies of peptide-membrane interactions constitute a challenging topic of research and complete understanding of the relationship between the structure of the peptide and the mechanism of interaction with lipids, as well as molecular details of this process, still remain elusive. However, it is of paramount importance to reveal the biological functions of membrane active peptides and to design peptides with tailored functionalities that may be exploited for therapeutic applications.For example, antimicrobial peptides (AMPs) are able to recognize and kill many pathogens, and a number of these peptides have been identified as key components of the natural immune defense system. A related family of peptides, the so-called cell-penetrating peptides (CPPs), is capable of efficient translocation across the cell membrane, either by themselves or together with a molecular cargo, and are being explored as potential programmable drug delivery vectors . There iMembrane interacting peptides can be classified according to their structure or their interaction with lipid membranes. They can assume different structures such as helical, β-stranded, mixed (containing both helices and strands) and cyclic, which are fundamental for modulating their membrane function. They usually contain amino acids with marked hydrophilic  or relatively hydrophilic  character, according to the Wimley and White hydrophobicity scale , which iAmong membrane interacting peptides, those that cause membrane alteration or permeation are fundamental because they may be exploited for the obtainment of potential new antibiotics. On the contrary, peptides that do not disrupt the membrane modulate the structure and dynamics of the lipid bilayer and also play key roles in protein trafficking, exo and endocytosis. Intensive research efforts have been carried out to elucidate the interaction of peptides with lipid bilayers at atomic detail, taking into account the position, orientation, structure, and dynamics of the peptide in the lipid bilayer and its effects on surrounding lipids. Therefore, all this information is necessary to unveil their functioning modalities and to envisage their potential applications.The complexity of the biological membrane is overwhelming and the multiple processes that occur simultaneously within this unique environment are just beginning to be understood.A variety of structural models describing interactions between peptides and membranes have emerged and have been investigated in recent years, such as the transmembrane channel aggregates, the “barrel-stave” model, and the “carpet” mechanism . The maiMany of such mechanisms include diverse stages of interaction between the peptides and the membrane or between various peptides, meaning that peptide-membrane interactions are complex and diverse phenomena. Depending on their composition, charge, and structure different peptides employ different interaction mechanisms with the membrane. In this review, we will focus on some of the most commonly seen scenarios in the studies of peptide-membrane interactions and on the different techniques that could be used to obtain deep details of the interactions.Peptide membrane interactions are important for the selective targeting of peptides to specific types of cells. Biological membranes are complex structures composed mainly of proteins and lipids which do not form a single homogeneous mixture, being some regions are more enriched in some components of the mixture. The membrane bilayer of different organisms can be very different. For example, the plasma membrane of bacteria and mammalian cells differ in their composition and properties and this accounts for specific interactions of peptides with one or the other membrane. The cationic characteristics of AMPs mainly contribute to cell selectivity, because the surface of bacterial membranes is more negatively charged than that of mammalian cells. The cell membranes of bacteria are rich in acidic phospholipids, such as phosphatidylglycerol and cardiolipin. The cell walls also contain anionic molecules, such as lipopolysaccharides in the outer membrane of Gram-negative bacteria and teichoic acids and lipoteichoic acids in the peptidoglycan of Gram-positive bacteria. In mammalian cells, acidic phospholipids are usually only present in the inner leaflets of plasma membranes, while the outer leaflets are mainly composed of zwitterionic phosphatidylcholine and sphingomyelin, although negatively charged gangliosides can also be present as minor species.Among the varied lipid components of mammalian membranes, the structure and physical properties of cholesterol  is rather different from that of other major membrane lipids; in fact, cholesterol is characterized by the presence of a hydrocarbon tail—a ring structure region with four hydrocarbon rings—and a hydroxyl group, and in lipid membranes, is slightly tilted with respect to the bilayer normal configuration, with the hydroxyl group positioned in the lipid head group region. This feature induces very different energies of interaction between proteins and cholesterol compared to other membrane lipids. In particular, depending on the temperature, cholesterol has different effects on membrane fluidity. At high temperature, it interferes with the movement of phospholipid fatty acid chains, making the membrane less fluid while at lower temperatures it prevents membranes from freezing and maintains membrane fluidity. These characteristics determine a segregation of components in membranes with the formation of cholesterol enriched microdomains (lipid rafts) . The facde novo designed and synthetically produced [http://www.bbcm.univ.trieste.it/*tossi/amsdb.html or at http://aps.unmc.edu/AP/main.php. AMPs have the ability to kill pathogenic microorganisms, including Gram-positive and Gram-negative bacteria, viruses, protozoa, and fungi; thus providing a first unspecific defense mechanism against microbial invasion. Moreover, they play an important role in the innate immune system of higher organisms  [AMPs were first discovered some decades ago in plants, insects, amphibian venoms and tissue extracts . Numerouproduced –23. An omammals) ,25, whicAMPs generally consist of less than 60 amino acid residues, and bear a positive net charge, which is often due to the Lys and Arg residues –31. AddiConsidering their secondary structure, AMPs can be divided into four different groups : (i) linAMPs display antimicrobial activity at micromolar concentrations or below. Besides membrane disruption, their bactericidal mode of action may involve interference with metabolic processes or targeting of cytoplasmic components . PredomiAs a consequence of their membrane target and mechanism of action, development of bacterial resistance towards AMPs is much less likely to happen compared to conventional antibiotics. Therefore, AMPs have been brought into focus as potential candidates for a new generation of antibiotics.Drosophila melanogaster Antennapedia proteins [Some intracellular proteins are able to pass through the cell membrane. Since the earliest discovery of this property for the HIV Trans-Activator of Transcription (Tat) and proteins ,37, manyA wide range of different applications of CPPs has been described, including transport of proteins, oligonucleotides, quantum dots, polysaccharides, nanoparticles, chemotherapeutics, polymers, and liposomes –47.Like AMPs, CPPs carry a positive net charge that is due to a large amount of basic amino acids (such as Arg and Lys) in their peptide sequence, and are also characterized by an amphipathic structure. The growing number of CPPs discovered so far is divided into natural, synthetic and chimeric peptides . The mecCPPs are overall hydrophobic, an essential characteristic for the interaction with the lipid part of the cell-membrane. The combination of a charged and a hydrophobic domain plays a key role in the interaction with natural membranes; the charged moieties are involved in the initial interaction with the bilayer while the hydrophobic domain is crucial for insertion.Most CPPs are amphipathic, consisting of two domains: a hydrophilic (polar) domain and a hydrophobic (non-polar) domain. Such an amphipathicity can occur at the levels of their primary structure or secondary structure. Primary amphipathic peptides correspond to the sequential assembly of a hydrophobic domain with a hydrophilic domain separated by a spacer domain; while secondary amphipathic peptides are generated by the conformational state which positions hydrophobic and hydrophilic residues on opposite sides of the molecule.All enveloped viruses share common steps of entry to the host mammalian cell using two main routes, either by the endocytic or nonendocytic pathways. Clathrin coated vesicles, macropinocytosis and caveolae are all pathways exploited by viruses preferring the endocytic route, while the non-endocytic pathway involves direct crossing of the plasma membrane at neutral pH. Regardless of the chosen route, the basic mode of entry by enveloped viruses is through membrane fusion, an essential and ubiquitous mechanism in most cellular events. Fusion is mediated by viral membrane proteins which undergo remarkable conformational modifications as a consequence of a trigger that is represented either by low endosomal pH or receptor binding. These conformational changes lead to the exposure of hydrophobic peptides, loops or patches , which then interact with and destabilize one or both of the opposing membranes. Three different classes of viral fusion proteins have been identified to date based on their common post-fusion structural motifs . These aHowever, the overall activity of the three classes of fusion proteins, regardless of their structural and biochemical differences, seems to induce membrane fusion in a similar manner. In fact, after fusion activation all fusion proteins will end up forming a similar hairpin structure. The principal element of the fusion machinery is always represented by a fusion peptide able to be inserted into cell membranes and drive membrane destabilization. Following activation by receptor binding or acidification of the endosomal compartment, the fusion peptide is projected toward the most external side of the glycoprotein where it is inserted into the target membrane. The structural changes in the fusion protein end up in the correct position of the target membrane, held by the peptide or fusion loops, and the viral membrane, held by the trans-membrane region of the fusion protein. Further remarkable refolding steps result in the merging of the two lipid layers and the consequent release of the viral nucleocapsid inside the host cells.The precise molecular events that lead to fusion are still unknown. Surely, the membrane leaflets have to be modified in order to allow accomplishment of fusion. It is now widely accepted that fusion peptides are capable of bilayer destabilization, by a combination of lipid head group charge neutralization and a deep localization of the peptide in the bilayer ,53. It iN-terminal region of VP1 of picornaviruses [In contrast, non-enveloped viruses, which by definition lack an outer membrane, mediate membrane fusion via utilization of previously sequestered viral lytic factors. The process of non-enveloped virus’ entry generally starts with the interaction with a cellular stimulus  at the penetration site which drives a conformational change that triggers the release of viral components with membrane lytic activity. This is followed by the binding of the small viral peptides or their hydrophobic domains with membranes to allow the transfer of viral genetic material and/or nucleocapsid to the cytoplasm. The molecular details of membrane disruption and the structural characteristics of the domains involved vary for different viruses. Examples of membrane altering sequences derived from several nonenveloped virus capsid proteins are: (i) peptides generated by autocatalytic cleavage of a precursor such as in the case of the γ peptide of nodaviruses ; the VP4aviruses ; and theaviruses ; (ii) peaviruses ; (iii) paviruses ; (iv) peaviruses ,62.Notwithstanding their origin, viral entry peptides are recently finding applications for the design of novel drug delivery tools due to their ability to cross membrane bilayers ,63,64.Amyloids are insoluble fibrillar aggregates of proteins/peptides . AccumulAmyloidogenic peptides are natively unfolded peptides which interact with lipid membranes and are involved in the formation of amyloid fibrils . The forIn vivo amyloid peptides have been shown to disrupt intracellular calcium regulation, plasma membrane potential, mitochondrial membrane potential and function and long-term potentiation in neurons. The formation of the β-sheet conformation from native protein structures can be induced by high protein concentrations, metal binding, pH, amino acid mutation and interaction with lipid membranes [Most of these peptides have been demonstrated to be capable of channel formation in lipid bilayers and it has been proposed that this may represent one of the pathogenic mechanisms. The channels formed exhibit a number of common properties, including irreversible, spontaneous insertion into membranes, production of large, heterogeneous single-channel conductances, relatively poor ion selectivity, inhibition of channel formation by Congo red and related dyes and blockage of inserted channels by zinc. There are also other possible mechanisms for membrane permeabilization which may involve interactions with specific membrane receptors  or distoembranes –80. Poreembranes . Recentlembranes ,82. Inseembranes ,12,83.The binding, location and orientation of a peptide relative to a lipid bilayer as well as lipid rearrangements in the presence of peptides are the most important features of peptide-lipid interactions. During the last decades, several experimental techniques have been developed and applied to biological systems, which differ in the nature and size of the sample, the sensitivity and the type and resolution of the information that they can provide. This section describes some of the most recent and significant methodologies which are usually combined in biophysical studies of membrane interacting peptides, since they provide complementary information .There are techniques that describe the morphological changes induced by peptides such as electron microscopy (EM), atomic force microscopy (AFM) and fluorescence imaging –87. EM aTechniques such as fluorescence, electron paramagnetic resonance spectroscopy (EPR), infrared spectroscopy (IR), circular dichroism (CD), surface plasmon resonance (SPR), X-rays and NMR illustrate interaction and are capable of describing both peptide and lipid structure and dynamics. The secondary peptide structure upon binding, degree of penetration and orientation can easily be obtained from IR, CD and EPR studies –103. NMRComputational methods have been widely used for studying peptide membrane interactions. Computer simulations are in fact an essential tool in the study of dynamic processes in biology and chemistry, providing data on sizes and time scales that are not accessible to experimental techniques. The mode of insertion of helical peptides has been predicted with success ,109, butExperimental techniques that have been used to get structural information include X-ray diffraction, Neutron Reflectivity methods (NR) as well as NMR, EPR, AFM.The possibilities of using X-ray diffraction to gain detailed insights into peptide-membrane systems are limited, because of the lack of long-range crystalline order. Crystal structures have been obtained only of a few membrane-bound peptides, which were crystallized from water or co-crystallized using organic solvents as membrane mimetics ,110.in situ measurements at solid-liquid interfaces can be performed. An important advantage of this method is the absence of sample damage, as frequently observed in X-ray reflectivity experiments, even upon prolonged exposure to the neutron beam. The information that can be extracted from a single neutron reflectometry experiment includes the film thickness, the scattering length density profile across the film and the surface roughness σ at the solid-liquid interface. Several applications to the study of membrane interacting peptides have been reported recently [Neutron reflectivity (NR) is a well-established technique for studying membranes at the solid/water interface, and provides detailed information on the structure and on the composition of the interface, but also of buried materials. Because of the large penetration depth of neutrons, due to their weak interactions with almost any material, recently ,112.13C, 15N and 2H [1H-15N bond relative to a molecular reference frame. The relative orientations of α-helices in the membrane can be determined, as well as any deviation of helices from their ideal geometry, like bends or kinks; the only information that cannot be obtained is the immersion depth. Another of the most frequently used solution NMR methods for investigating the orientation and localization of peptides bound to membrane-mimetics uses the effect of paramagnetic probes on the peptide itself or tags on the peptide [NMR—and specifically, solid state NMR—has numerous applications in peptide membrane interaction studies; in fact, a series of NMR experiments have provided important insights about peptide orientation ,113–117.N and 2H . A varieN and 2H ,118–121.N and 2H –124. MicN and 2H . AnotherN and 2H . The RDC peptide . For exa peptide . The int peptide . In solu peptide . Two difEPR has been widely used to monitor membrane interactions ,131–134.AFM has also been used for the characterization of peptide-membrane systems –87. DestThe understanding of the interaction of peptides with lipid bilayers at atomic level requires characterization of the position, orientation, structure, but also dynamics of the peptide in the lipid bilayer and its effects on surrounding lipids. Molecular dynamics (MD) simulation is a powerful research tool to theoretically study peptide-membrane interactions, which can provide a detailed description of these processes at a molecular level. However, considerable obstacles need to be faced to obtain accurate results, due to inexact force fields and other approximations in simulation methodology, and to the relaxation times of these phenomena, which are typically longer than those presently accessible to MD. In fact, in order to fully describe peptide-membrane interactions, it would be necessary to consider the behavior of an individual or several peptide molecules, to elucidate the dynamics of the peptide-membrane assembly process, to provide a link between the observed behavior and the data measured in experiments and to systematically explore a large number of systems.Over the years, a number of theoretical and computer simulation approaches have been developed to describe membrane behavior and peptide-membrane interactions, which vary in the way the peptide-membrane system is modeled, and thus, in the type of information that can be obtained from each particular model ,139–143.Several atomistic molecular simulation studies attempted to address long scale peptide-membrane interactions in their full complexity showing pore formation ,144, pepThese limitations in time and length scales led to the development of coarse-grained approaches (CG) for the study of complex biomolecular phenomena, in which the time problem may be circumvented at loss of resolution . CG apprRecently, the CG models have been further extended, representing each amino acid according to its properties, such as tendency to form hydrogen bonds, hydrophobicity/hydrophilicity and charge, or representing the backbone of the amino acid and the side chains as different beads –152. MorThere are also other experimental techniques that can be used in order to map the position of different molecules in a peptide-lipid bilayer system.Fluorescence spectroscopy is widely used for determining the approximate position and orientation of lipid associated peptides and membrane affinities. Both the peptide and/or the surrounding lipids could be labeled with a fluorescent tag. This technique also allows investigation of the fast kinetics of peptide insertion into the membrane. Membrane bound peptides often contain, or are modified to contain, a tryptophan (Trp) residue which possesses intrinsic fluorescence. Moreover, Trp is extremely sensitive to the polarity of its surrounding and can be used for calculating binding affinities for membrane-mimetics. Upon interaction with a hydrophobic environment, the Trp fluorescence emission is shifted to shorter wavelengths (blue shift) and decreases in intensity ,155,156.Surface plasmon resonance (SPR) has become one of the most important techniques for studying macromolecular interactions. Recent advances in the preparation of stable membrane-like surfaces and the commercialization of sensor chips have enabled widespread use of SPR in the study of membrane interactions. The number of published papers has increased steadily in the last few years, especially since Biacore introduced the HPA and L1 chips, dedicated to lipid systems ,163–169.SPR studies, in general, provide both qualitative and quantitative data on molecular interactions .The typical qualitative study allows the determination of lipid specificity of a membrane-binding peptide, which very often is the key regulatory step of peptide action. Lipid specificity can be easily studied by manipulating the lipid composition of the immobilized membrane. The visual inspection of binding curves can immediately deliver qualitative information to describe how peptides bind to some kinds of lipids. Moreover, it is possible to determine the apparent rate and affinity constants from sensorgrams, especially when the differences between various conditions, either different types of membranes/buffers/pH or different variants of systems studied, are subtle. The affinity constants can be directly determined from the equilibrium binding responses over a range of peptide concentrations, by fitting the data to a Langmuir adsorption isotherm. In addition, binding constants can be determined directly from the sensorgrams by numerical integration analysis –173. ThiKAbilayer/KAmonolayer provides an indication of the depth of penetration into the membrane core. A ratio of ~1 indicates that the peptide is surface localized, and a ratio above ~10 indicates that the peptide has been inserted into the lipid bilayer. Several studies have reported this behavior, using calculations of different binding affinities of peptides on L1 or HPA chips [Additional information about the mechanism of membrane association can be obtained when both HPA and L1 chips are used. Due to structural differences between the two chips, it is possible to distinguish between surface adsorption in the HPA chip and insertion into the hydrophobic core of the membrane in the L1 chip. If the peptide binds only to the water–lipid interface, and binding is not accompanied with deeper penetration, then similar equilibrium constants can be observed with both chips. On the other hand, if the peptide inserts deeper in the membrane and needs a transmembrane compartment, then no binding is observed in HPA. A difference between the affinities of the peptides to lipid monolayers, compared to bilayers, indicates the contribution of the membrane hydrophobic core to the binding process. The ratio PA chips –174. In Isothermal calorimetry (ITC) is another methodology which has proved to be very useful for the study of peptide membrane interactions. The development of high sensitivity titration calorimeters has allowed the measurements of binding reactions with solutions in the micromolar concentration range. A complete thermodynamic picture can be obtained to provide information about the driving forces of the binding reaction. ITC can also be used to investigate secondary processes accompanying peptide-membrane interactions, such as membrane permeabilization, peptide induced lipid phase transitions, peptide aggregation at the membrane surface and peptide conformational changes .The phase transition temperature and thermodynamics of lipid phase transitions are extremely sensitive to the presence of exogenously added compounds. Differential scanning calorimetry (DSC) has been used in a variety of peptide lipid studies to monitor changes in these parameters and obtain valuable information regarding the ability of peptides to interact and/or disrupt the lipid acyl chain packing also providing insight into their interaction mechanism ,175,176.CD spectroscopy is used to study the secondary structure of peptides and its changes in different environments such as temperature, pH, ligands, or denaturants . The oriThe interaction of peptides with membranes is a topic of great interest, both for the purpose of understanding the mechanism of action of membrane-active peptides as well as for designing peptides which can modulate membrane properties and may have applications for human health .The great interest in biophysical studies of the interaction between AMP and lipids is in part due to the rapid emergence of antibiotic-resistant bacterial strains. The hope is that the understanding of the molecular mechanisms used by AMPs for membrane perturbation may allow the design of novel peptide antibiotics which could be used as an alternative or in combination with conventional antibiotics. The biological activity of AMPs stems from their ability to perturb the lipid bilayer structure of membranes. Moreover, some of this peptides exhibit high specificity towards their target membrane and are toxic to humans while others are toxic only to microorganisms. The understanding of the damaging properties on specific target membranes plays a key role in the rationale design of novel peptide antibiotics, and is thus pharmaceutically relevant.The transport of drugs or drug delivery systems across the cell membrane is another complex biological process, often difficult to measure because of its dynamic nature. Biophysical investigations play a critical role in understanding the mechanism of cellular uptake, the toxicity of drugs and to optimize drug delivery systems . Model mMembrane translocation is a necessary step for membrane permeabilization, and in fact, CPPs and AMPs are very similar molecules, and although being treated differently, both interact with the bilayer. At high concentration, peptides reported as CPPs perturb the membrane and become membrane permeabilisers, whereas at low concentrations, AMPs may reach cytoplasmic targets before membrane permeabilization. Cellular membranes of target cells where the activity of these two classes of peptides are evaluated are quite different. In fact, CPPs are evaluated against mammalian cells, whereas the target of AMPs is the bacterial cell. The bacterial membrane has a higher percentage of negatively charged lipids and does not contain sterols, thus, different effects reported with CPPs and AMPs can arise from the differences in membrane composition, and other factors which modulate peptide affinity and membrane destabilization. Another class of membranotropic peptides that is an object of active research for the design of novel delivery tools is represented by viral-derived peptides.The membrane affinity and the capacity to destabilize the bilayer dictate the extent to which a peptide enters the cell by a physical mechanism at the expense of the endosomal pathway. Most CPPs induce an endocytotic mechanism with the first step of their cellular entry being the electrostatic interaction with the cell surface. In particular, conformational changes modulate peptide/lipid interactions and the strength of these interactions might determine whether the peptides remain entrapped at the plasma membrane or not and whether they follow an endocytotic route or induce membrane disorganization with direct translocation. In this context, it is of paramount importance to introduce novel drug delivery peptides which use mainly a translocation mechanism to enter cells such as viral membranotropic peptides, which have not been yet widely exploited. Moreover, it is particularly intriguing the use of Class I viral fusion peptides as drug delivery vectors, as they are characterized by a structural plasticity which seems to be fundamental for membrane interaction and their hydrophobicity together with the presence of a few charged residues allows both electrostatic and hydrophobic interactions which support a translocation mechanism through the membrane bilayer.Delivery across cellular membranes involves several mechanisms such as direct transfer through cell surface membrane by lipid membrane fusion or transient permeabilization of the cell membrane; or after endocytosis, transfer across vesicular membranes by lipid disruption, pore formation or fusion. Several of these membrane reorganization steps are involved in the entry of viruses or other intracellular microorganisms, and are also triggered by protein toxins and defense peptides ,5,12. ReTwo fusion peptides belonging to Class I have been so far exploited as delivery vectors, in particular, the influenza and the HIV fusion peptides –188. TheC-terminus; when the arginine is mutated the fusogenic activity of the peptide is strongly impaired. The hydrophobic domain is also crucial for insertion of the peptide in the membrane and corroborates the notion that hydrophobic interactions between fusion proteins and cell-membrane phospholipids initiate membrane perturbation in the early stages of viral fusion. gH625 cellular uptake is thus associated with its ability to interact with membrane lipids and to form a transient helical structure that temporarily affects membrane organization, thereby facilitating insertion into the membrane and translocation.MPG is a chimeric peptide derived from the fusion peptide of HIV-1 and the nuclear localization sequence of SV40 large T antigen ,187. TheN-methyl amino acids [Many amyloid diseases are neurodegenerative and terminal and affect large portions of the elder population, thus prompting massive scientific interest for research on inhibition of any step of amyloid formation –202. Sevno acids , prolineno acids  or sugarno acids  have proThe exact manner used by peptides to interact with membranes and molecular details of this process are still an area of active research and a matter of extensive debate and controversy. Different peptides utilize different interaction mechanisms or combinations of mechanisms. Furthermore, the mechanisms of interaction can change depending on the conditions of the system, such as pH, temperature and concentration of peptide.In order to understand the biological processes of peptide-membrane interactions, or to design peptides with tailored functionalities for specific applications, we need to establish, with molecular resolution, the link between the structure and physical characteristics  of the peptide and the particular interaction mechanism it induces.It is clear that a better understanding of peptide-membrane interactions at a molecular level not only is important in the elucidation of various biological processes, but also could be instrumental in designing peptides with tailored functionalities, for example, for antibiotic and drug delivery applications.


Bangladesh has a high proportion of households incurring catastrophic health expenditure, and very limited risk sharing mechanisms. Identifying determinants of out-of-pocket (OOP) payments and catastrophic health expenditure may reveal opportunities to reduce costs and protect households from financial risk.This study investigates the determinants of high healthcare expenditure and healthcare- related financial catastrophe.A cross-sectional household survey was conducted in Rajshahi city, Bangladesh, in 2011. Catastrophic health expenditure was estimated separately based on capacity to pay and proportion of non-food expenditure. Determinants of OOP payments and financial catastrophe were estimated using double hurdle and Poisson regression models respectively.On average households spent 11% of their total budgets on health, half the residents spent 7% of the monthly per capita consumption expenditure for one illness, and nearly 9% of households faced financial catastrophe. The poorest households spent less on health but had a four times higher risk of catastrophe than the richest households. The risk of financial catastrophe and the level of OOP payments were higher for users of inpatient, outpatient public and private facilities respectively compared to using self-medication or traditional healers. Other determinants of OOP payments and catastrophic expenses were economic status, presence of chronic illness in the household, and illness among children and adults.Households that received inpatient or outpatient private care experienced the highest burden of health expenditure. The poorest members of the community also face large, often catastrophic expenses. Chronic illness management is crucial to reducing the total burden of disease in a household and its associated increased risk of level of OOP payments and catastrophic expenses. Households can only be protected from these situations by reducing the health system's dependency on OOP payments and providing more financial risk protection. The fundamental role of a healthcare system is not only to improve population health but also to protect households from financial catastrophe associated with illness Like many developing countries, Bangladesh is also facing the double burden of disease Thus, previous estimates may have provided an incomplete picture of the impact of medical expenses on populations with a high prevalence of chronic illness and limited risk-pooling mechanisms. In designing healthcare financing systems, policy makers need to understand determinants not just of OOP payments, but also the related problem of catastrophic health expenditure associated with high OOP payments The study was conducted in Rajshahi city, Bangladesh, which is the third largest city in the country and broadly representative of many urban areas in Bangladesh Mahallah, the lowest administrative region of a Bangladeshi city. In the first stage, 40 clusters were randomly selected from 159 eligible PSUs with probability proportional to size. In the second stage, a fixed number (40) of buildings was selected by systematic random sampling from each chosen cluster based on a household listing operation to provide the necessary frame for selecting buildings. During the final stage, one household was randomly selected from each building, with a target sample of 1600 households. Of these households, only seven refused interview or were not available to be interviewed, and the final effective sample size was 1593 households, resulting in a response rate of 99.6%.A cross-sectional three-stage cluster sampled household survey was performed during August to November 2011. The primary sampling unit (PSU) was the Respondents were administered a structured questionnaire developed based on the Bangladesh Household Income and Expenditure Survey (HIES) The respondents in this study were the women in the household, the household head, or the most knowledgeable person in the household, where necessary. Informed consent was obtained prior to conducting the interviews. The survey questionnaire contained two main sections: the household questionnaire and the individual illness questionnaire. The household questionnaire covered survey and household identification and household consumption expenditure, including food consumption, non-food expenditure, housing and durable goods; the individual questionnaire contained demographic information such as age, sex, marital status, education and occupation of individuals in the household, and health problems in the past 30 days. The food consumption section covered purchased, home-produced and in-kind consumption in the past 30 days or past 12 months prior to interview. Following the same recall process as food consumption, the non-food expenditure section also covered purchased and in-kind goods. Housing rent or equivalent rents were recorded in the past 30 days and the durable goods section recorded detailed information on number of items, duration, present and past value of the most recent items in the one year recall period. Cost of medical expenses including fees , drugs and medical supplies, transport costs for patients and accompanying family members, and other costs were recorded for each episode of illness in the past 30 days prior to interview. All expenditure was recorded in the Bangladeshi currency, taka (TK). In addition, data on timing and cost of all episodes of illness, care-seeking behavior and inpatient or outpatient care were recorded for the past 30 days prior to interview.Consistent with common definitions, OOP healthcare expenditure was defined as ‘catastrophic’ if it exceeded 40% of household non-food expenditure or capacity to pay in the past 30 days Descriptive statistics were calculated using the mean , median (inter-quartile range) or frequency and proportions as appropriate. Trend tests were performed using the Mantel-Haenszel chi-square test for categorical variables and linear regression analysis for continuous variables, with ordinal numbers (1–5) assigned to the quintile categories. Double hurdle and Poisson regression models were used to identify the determinants of OOP payments and catastrophic expenditure, respectively. A brief description and motivation of the models is given below.Reporting of zero expenditure is quite common in household consumption expenditure surveys. For example, both medical and tobacco consumption expenditure are zero for many individuals or households over a survey recall period. In addition, participation in expenditure and the magnitude of expenditure may not be statistically independent The double hurdle model was used to assess the relationship between demographic and household variables and the size of OOP expenses. The dependent variable for the probit model is a dichotomous variable that indicates whether OOP expenses were incurred (the participation decision). The Tobit regression model analyses the natural logarithm of OOP payments as a function of the covariates (the consumption decision). This model can be presented symbolically through two related equations for participation and consumption.* is another latent variable describing household level of healthcare expenditure, y is the observed dependent variable , z is a vector of variables explaining the participation decision, and x is a vector of variables explaining the expenditure decision. According to Jones, the likelihood function can be written as p then denotes probability of expenditure, Observed consumption:kelihood .In the case of rare events, Poisson regression can provide more accurate estimates than logistic regression All analyses at both the univariate and multiple regression stages were adjusted for the probability sample design. Statistical analysis was performed using Stata/SE Version 12.0.The study modeled households' OOP health payments and risk of catastrophic expenditure as a function of household characteristics and economic status and presence of illness and care-seeking behavior, using average illness per child and adult as a measure of illness This study received ethical approval from the Ethics Committee of The University of Tokyo and the Bangladesh National Research Ethics Committee, with reference number BMRC/NREC/2010-2013/1161. About one third of the population in Rajshahi city are still illiterate and even written consent is not common practice among them. Therefore, a consent form to obtain and document verbal or written consent from respondents was proposed and approved by the Ethics Committee together with the study protocol. Prior to the interview, our enumerator carefully read the consent form to the subject and then very briefly explained the aims and importance of the study. This consent form contained information on the objectives of the study, risks, benefits and freedom of participation, and confidentiality.The incidence of catastrophic healthcare expenditure by illness, care-seeking behavior and household level characteristics is presented in 2(1) = 8.88; p<0.01), indicating a double hurdle model is appropriate for this data. Presence of chronic illness, household size, average illness per child and adult, care-seeking behavior, education level of the household head and household consumption quintile significantly affected the level of household OOP healthcare spending.Results of the double hurdle model are presented in This paper, based on a representative household survey in Rajshahi city, Bangladesh, is the first to consider illness, care-seeking behavior, demographics of the household head, and household economic characteristics as household-level predictors of OOP payments and catastrophic expenditure. It is also among the few examples of studies that have reported the incidence of catastrophic healthcare expenditure in Bangladesh This study found that sampled households, none of whom have any form of risk-pooling insurance, spend about 11% of their total household budget on healthcare, and nearly 9% of households experience financial catastrophe. At a 25% non-food expenditure threshold, the incidence of financial catastrophe was similar to another published study, at 18% This study revealed that the per capita monthly OOP health expenditure made by households was TK 138.0 (US$ 1.8), which is similar to national-level findings in Bangladesh Consistent with other studies Our analyses demonstrate a negative impact of average illness per child and adult, and presence of chronic illness in the household, on the household economy. These results are similar to the determinants of catastrophic expenditure in Burkina Faso and India Type of health service used was also another important determinant of OOP payments and financial catastrophe, with intensity of OOP payments at public outpatient facilities lower than private outpatient facilities. These findings are similar to several studies from developing countries The research protocol and sampling process in our study was designed to avoid any biases in the results, but our study has several limitations. We examined only urban households in one metropolitan area of the country, so the results cannot necessarily be generalized to the whole country. However, the representative nature of the sample means that the results may be applicable to other cities, and thus the study may reflect the reality of health market participation for a large proportion of the Bangladeshi population. Inpatient service use is infrequent (4%) and a much larger sample is required to explore the role of chronic vs. acute illness in hospitalization and costs. Such an analysis might better describe the role of preventable hospital admissions in catastrophic spending. Consumption and expenditure were self-reported and prone to error, although estimates were confirmed by other household members or aged persons in the community. For example, female interviewees frequently over- or under-estimated the cost of bicycles, sewing machines and cars, but we minimized the bias by asking another household member or an older member of the household.This study identifies determinants of high medical expenditure and financial catastrophe: illness either in children or in adults, chronic illness, receiving inpatient care, poorer economic status and lower education level of the household head. The chronic care of NCDs requires long-term routine clinic visits, testing, and medications, reducing households’ flexibility to respond to the cost of unexpected hospitalization or other illness episodes. It is clear that immediate action is necessary to reduce levels of catastrophic health expenditure by reducing the burden of OOP payments in Bangladesh, which can be achieved by:Implementing compulsory health insurance for salaried workers in both public and private sectors, and voluntary memberships for dependents, farmers and self-employed persons, similarly to programs in Vietnam that have been shown to reduce OOP payments to lower levels than observed in this study Improving routine management of NCDs, to reduce the cost of chronic disease management, and incorporating chronic disease management into public services and health financing initiatives, to ensure that this expenditure is included in risk-pooling and welfare initiatives and the high OOP payments associated with chronic illness that were identified in this study can be ameliorated by better and more equitable management, prevention and treatmentIncorporating ancillary services into basic care packages in public facilities, so that users are not required to pay significant OOP expenses for essential pharmaceutical or other ancillary services which are supposed to be almost free, but which our study found were still associated with high OOP payments and catastrophic expenditure riskIf necessary reforms are implemented, especially those targeted at the poorest members of Bangladeshi society, significant reductions in the burden of OOP payments can be made, with consequent improvements in the health of the population.


Verrucous carcinoma has a special propensity to mimic benign lesions of the oral cavity. A case of the oral verrucous carcinoma in maxillary alveolar ridge, extending to buccal vestibule, cheek, and labial mucosa, which was diagnosed and initially treated as chronic candidiasis, is presented. Clinical, histopathological, and therapeutic considerations related to diagnosis of the verrucous carcinoma in the oral cavity are discussed. The verrucous carcinoma, a low-grade variant of squamous cell carcinoma, has been reported in the head and neck area, with predilection by oral cavity and larynx –4. The cThe establishment of clinical or histopathological diagnosis of verrucous carcinoma in the oral cavity may be difficult to interpret. This difficulty has been attributed to the indolence of this tumor's grow, its benign histologic features, and the lack of awareness by the pathologist of the tumor's gross appearance , 4.We present the case of a patient with oral verrucous carcinoma in maxillary alveolar ridge, which was diagnosed and initially treated as chronic candidiasis. Clinical, histopathological, and therapeutic considerations related to diagnosis of the verrucous carcinoma in the oral cavity are discussed.A 67-year-old female was referred for evaluation of verrucous white lesion in the oral cavity. There was no history of tobacco or alcohol consumption. According to the patient, the lesion was present for approximately 24 months and it had a slow growing in the last six months. She had been submitted to an incisional biopsy whose diagnosis was of chronic candidiasis. As a result, antifungal therapy (Fluconazol 150 mg/week) was provided during six months without any remission of the lesion.Clinical examination revealed the presence of a wide verrucous white lesion in the molars region of the maxillary alveolar ridge, extending to buccal vestibule, cheek, and labial mucosa . The subRadiographs examination showed no infiltration of the maxillary bone. The differential diagnosis included verrucous leukoplakia, verrucous carcinoma, and squamous cell carcinoma. Then, two incisional biopsies were performed in different areas of the lesion and submitted to histopathological analysis.Microscopic features showed stratified squamous epithelium with intense hyperplasia, hyperparakeratosis, low atypia degree and formation of keratin pearls . The retGenerally, the verrucous carcinoma occurs in elderly and tumors of the tongue, gum and buccal mucosa are more common among females . The etiThe diagnosis of the verrucous carcinoma may be difficult, requiring, in some cases, several biopsies. This lesion has a special propensity to mimic of benign tumors of the oral cavity. The clinical and histopathological differential diagnosis should include pseudoepitheliomatous hyperplasia, well-differentiated squamous cell carcinoma, chronic candidiasis, and condyloma accuminatum. The definitive diagnosis obviously requires concurrence between the clinician's appreciation of the tumor and the pathologist's identification of the microscopic criteria described by Ackerman , 6. The The treatment of oral verrucous carcinomas, as in the reported case, consists of surgical excision without the need for lymph nodes dissection, because regional metastasis is rare , 4, 6, 9Finally, it is important to reinforce that appropriate diagnosis of the verrucous carcinoma in the oral cavity can prevent the most extensive involvement of adjacent areas and/or wide surgical resection of the lesion including lymph nodes dissection as recommended for some cases of oral well-differentiated squamous cell carcinoma.


In genome science, the advancement in high-throughput sequencing technologies and bioinformatics analysis is facilitating the better understanding of Mendelian and complex trait inheritance. Charting the genetic basis of complex diseases – including pediatric cancer, and interpreting huge amount of next-generation sequencing data are among the major technical challenges to be overcome in order to understand the molecular basis of various diseases and genetic disorders. In this review, we provide insights into some major challenges currently hindering a better understanding of Mendelian and complex trait inheritance, and thus impeding medical benefits to patients. Single-gene mutations in the form of rare clinical phenotypes and Mendelian disorders are being identified in every age and sex group of the human population . After cRb1 gene affected with retinoblastoma (OMIM: 180200) and not the other – although both carry the same mutation (NF1) gene  gene . After sF1) gene , environF1) gene , allelicIn addition to monogenic disorders , understanding the genetic basis of complex modes of inheritance relating to diseases of digenic, trigenic, and even more complex traits, is another challenge in genome science. In analyzing complex modes of inheritance, genome-wide association study (GWAS) strategies have not lived up to earlier expectations for the discovery of complex traits, due to the erroneous assumption that common disorders are regulated by common genetic risk factors . The sysPopulation genetic studies are important for understanding the distribution of genetic variability among populations, inferring their demographic histories and their adaptation to natural selection and genes flow, associated with human diseases and health . MoreoveToday, clinicians are faced with the difficulty of choosing between disease-targeted sequencing tests and high-throughput sequencing (genome and exome sequencing) approaches . Within In developed countries, cytotoxic chemotherapy and radiotherapies have raised the overall success rate in the treatment of pediatric cancer to 80%. However, a great reduction in life quality due to major side-effects of therapies is limiting the success of therapy –23. DuriThe presence of structural variations in repetitive DNA sequences, heterogeneity within tumor samples, and mutations affecting the regulatory regions of genes are the main foci of PCGP’s aim of better understanding the complications of genome data interpretation. The human reference genome still carries gaps and there is an incomplete picture of the full set of genome variations in human population groups . Hence, The striking degree of diversity in human tumor samples has resulted in the initiation of a number of different cancer projects aimed at developing the accurate estimation of cancer cells intermixing with normal support and immune cells . FinallyOver the last decade, numerous advances in the fields of structural and functional genomics have taken place . High-thde novo assembly of short reads, at repetitive loci, as delivered by most NGS technologies, is challenging. In addition to creating gaps, complex and misassembled rearrangements could be resulting from the erroneous collapse of repeats on one another. Consequently, the accurate and scalable assessment of structural variations solely by NGS platform is still problematic.  technology has made the problem of data analysis more challenging by generating substantial data in the form of whole-genome sequencing, exome sequencing, transcriptome analysis, expression profile chromatin immunoprecipitation-based (ChIP) sequencing, and methylome (epigenetics) . In NGS,lematic. . Despite problem , 29. AftRecent progress in acquiring complete genome sequence information is leading toward a personalized medicine tradition. Although whole genome and exome sequencing strategies are contributing to anticipate clinically significant variants, these are still in infancy. The accurate understanding of factors influencing rare Mendelian and common genetic traits, and the precise handling of high-throughput next-generation data, are the most significant technical challenges still impeding the medical benefits to patients. However, the cost-effective application of NGS technologies – quick, secure, accurate analysis of large scale NGS data – for identification of common and rare genetic variants across the human populations and diseased individuals together with advancement in the fields of proteomics and metabolomics will provide a way of knowledge transfer from scientific literature toward clinical practices.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.


When compared with Thailand, the seroprevalence of hepatitis A virus (HAV) is extremely high among its neighbouring countries. To investigate the seroprevalence of HAV among the Thai people residing in the border area between Thailand and Myanmar, 308 residents in Umphang, Maesod district, Tak, were recruited. Sera were tested for HAV IgG antibodies by enzyme-linked immunosorbent assay. The overall seroprevalence among the Thai people residing in the border area of Thailand was significantly higher than that among the general Thai population . As asymptomatic or mild HAV infection typically occurs in children, the Thai people residing in the border area may receive little benefit from universal HAV vaccination. Lower protective antibodies against HAV, along with the exclusion of HAV vaccine from the Expanded Programme on Immunization, potentially increase the susceptibility to HAV among the general Thai population and may lead to more future outbreaks if HAV is introduced from the border areas. The findings suggest that HAV vaccines should be recommended to travellers before their journey to the border between Thailand and Myanmar where HAV is endemic. The seroprevalence of hepatitis A virus (HAV) in most parts of Asia has gradually declined as a result of significant improvements in hygiene and living standards -3. LeastThe seroprevalence of HAV among the Thai people residing in the border area adjacent to Myanmar is yet to be studied, although, due to geographical proximity, it is presumed to be higher than that among the general Thai population. The Expanded Programme on Immunization (EPI) in Thailand has excluded HAV vaccine based on cost-effective analysis, indicating that the low incidence of HAV infection, along with the cost of the vaccine, tempers its cost-effectiveness . Yet, thThe participants included either healthy children who had attended the well-child clinic or patients with acute illness who had attended the outpatient clinic or had been hospitalized at the Umphang Hospital, Tak, Thailand, during January 2009–June 2010.oC until tested for anti-HAV by a commercial ELISA  at the Center of Excellence in Clinical Virology, Chulalongkorn University, Thailand. The seroprevalence of HAV in each age-group was calculated and reported as percentage. The age-related seroprevalence of HAV among the people residing in the border area was compared with that of Thai , which had a high seroprevalence of 60%. By the age of 40 years, almost every subject had developed HAV IgG antibodies.When comparing the seroprevalence of HAV among the people residing in the border area with that of Thai  and Myanet al. (HAV remains a major public-health problem wherever hygiene and sanitation are inadequate. Accordingly, the border area of Thailand is prone to sporadic outbreaks of HAV as previously reported in a study by Barameechai et al. . The serThe age-related anti-HAV seroprevalence among the people residing in the border area between Thailand and Myanmar was in between that observed among the Thai population and Myanmar immigrant workers in previous studies. The seroprevalence in all three populations increased in relation to the age of the subjects and reached its peak with the Thai people residing in the border area who, on average, were older than Myanmar immigrant workers but younger than positive subjects within the general Thai population. As it has been previously shown that HAV IgG antibodies can be trans-placentally transferred , the spiThe results of the present study reflect the hygiene and sanitation conditions in the border area of Thailand which may have had relatively little improvement when compared with the other parts of the country. It was reported that HAV infection in children usually exhibits mild non-specific symptoms and low case-fatality compared to HAV infection in adults , 10. FigThe authors express their deep gratitude to the Commission on Higher Education, Ministry of Education, Department of Medical Sciences, Ministry of Public Health, and Center of Excellence in Viral Hepatitis, Thailand Research Fund, Center of Excellence in Clinical Virology, Chulalongkorn University, CU Centenary Academic Development Project, and King Chulalongkorn Memorial Hospital for their generous support. Finally, they thank Ms Petra Hirsch for reviewing the manuscript.


The Lateral Habenula: No Longer Neglected (Geisler and Trimble, The habenula is a small, bilateral brain structure located at the dorsal end of the diencephalon. This structure sends projections to the dopaminergic striatum and receives inputs from the limbic forebrain, making the habenula a unique modulator of cross-talk between these brain regions. Despite strong interest in the habenula during the seventies and eighties (Herkenham and Nauta,  The habenula  is a small, complex, and evolutionarily conserved structure. The habenular complex on the dorsal diencephalon is surrounded by the third ventricle and includes the medial habenula (MHb) and lateral habenula (LHb) Figure . Both stThe MHb primarily receives inputs via the stria medullaris, a white matter pathway originating in the limbic system  via the internal portion of the fasciculus retroflexus (FR) , attention deficits, hyper-reactivity to stress, and schizophrenic-like symptoms  on the Morris Water Maze. These results prompted researchers to speculate about the possible involvement of the habenula in schizophrenia  receives the most scientific attention  injection (De Biasi and Salas, Another avenue of MHb research utilizes pharmacological agents that specifically target the habenula. The African shrub Tabermanthe iboga contains ibogaine, an alkaloid with hallucinatory properties. ibogaine has a number of anti-addictive properties and has been shown to block rodent self-administration of cocaine (Cappendijk and Dzoljic, ex-vivo brains, but not functionally in vivo (Strotmann et al., A few human neuroimaging studies have defined habenular activity (Ullsperger and von Cramon, This review addresses the importance of the MHb in regulating behavior and the involvement of this structure in multiple neuropathologies and addiction. A growing literature suggests that the MHb plays an important role in mood disorders, anxiety, stress, memory, and nicotine withdrawal as well as in cocaine, methamphetamine, and alcohol addiction. We strongly suggest that the MHb should no longer be neglected.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.


Rationale: There is little knowledge regarding mullerian adenosarcoma and its metastasizing pattern.Aim: Our objective was to evaluate the impact on the prognostic of the patient brought by tumor metastasizing to the retroperitoneum, to analyze the particularities of the treatment in such cases and to bring a significant change in the early therapeutic attitude to mullerian adenosarcomas.Methods and results: We present a first case report of a hypervascularized retroperitoneal metastasis from an initial low-grade uterine adenosarcoma. The presence of such a metastasis brought the worst prognostic factor to the patient. Conclusions: We consider that in front of a macroscopic polypoid mass there should be an active change in the diagnosis and in the therapeutic attitude. The frequent confusion between similar histopathological entities with different aggressiveness states and specific treatment responses, the poor outcome at advanced tumor stages, frequently in young patients, should trigger a universal remodeling in approaching these tumors. Their frequent confusion with recurrent cervical polyps or with other similar histopathological entities makes their diagnostication even more difficult, impeding an adequate precocious treatment. Mullerian adenosarcomas have been first described by Clement and Scully in 1974, but, due to their rarity, they still represent an elusive aspect of our knowledge. There is still a limited, small number of reports concerning these tumors, randomized trials have not been possible and screening methods are absent [2]. Typical mullerian adenosarcomas display a mixed composition: a benign neoplastic glandular component, with a well-differentiated epithelial lining, and a low-grade, malignant sarcomatous stromal structure. Usually, uterine adenosarcomas arise in the endometrium and more seldom within the myometrium. Rare cases of mullerian adenosarcoma of the cervix (4 reports in literature), vagina, ovaries, salpynx, or even more unusual extragenital primary mullerian tumors  have been described [5]. The majority of the uterine adenosarcomas are confined to the endometrium, but in 15% of cases, they can invade the inner half of the myometrium. The recurrences (in 20-30% of cases) are limited to the vagina, pelvis or abdomen and can appear after a long period of time. The treatment consists of a total hysterectomy with bilateral oophorectomy, but a more conservative attitude is followed in young women. There is no consensus regarding the complementary treatments [6]. Here, we present a very rare case of an aggressive retroperitoneal metastasis from a low-grade typical uterine mullerian adenosarcoma. To the best of our knowledge, there are no such reports in literature, the metastasizing rate being very low. Having this case in view, we consider that the current diagnosis and treatment of uterine tumors should be remodeled.Among the rare uterine sarcomas , mullerian adenosarcomas are even less observed and diagnosed, representing only 8% of all uterine mesenchymal malignancies . At low power magnification, adenosarcoma exhibits a leaf-like pattern, resembling phyllodes tumor. The glandular epithelium is active, like a proliferative endometrium, with mitotic activity, despite the advanced age of the patients, and with focal atypia; the stroma is composed of spindled cells arranged in whorls and/or loosely dispersed round cells. Stromal cells form a periglandular cuffing, a zone of maximum mitotic activity and nuclear atypia. Routinely, the diagnosis is made without a stromal cell mitotic count (that in adenosarcoma is more than 1-2 per 10 HPFs), immunohistochemical determinations  or DNA aneuploidia/diploidia determination. The features of an aggressive variant, the presence of sarcomatous overgrowth would change the specificity of the medical treatment and follow-up. We consider that in the case of uterine sarcoma, these determinations should be done, together with an analysis of the entire surgical resection piece and the abandon of a fast diagnosis in front of an apparent benign polyp. In the case presented by us, the discovery of a simultaneous ovarian metastasis at the time of the initial operation by histopathological examination, without a macroscopically altered aspect, suggests that maybe, the attitude of conserving the ovaries in younger women is not of the safest and should be, maybe, reconsidered. In this case, the initial tumor presentation displayed many of the characteristic features , with some factors suggesting a poor prognostic: concomitant metastasis, treatment and follow-up negligence, but also some encouraging: lack of sarcomatous overgrowth, a limited myometrium invasion, lack of risk factors. There still remains a question about the spectacular disappearance of the local recurrence after radiotherapy. But, even more unusual is the metastasizing of the tumor to the retroperitoneum that brought the worst possible prognosis to the patient. Usually, retroperitoneal tumors exhibit a silent growth until they reach gigantic dimensions, with multivisceral and large vessel involvement. Although their treatment is dependent on a radical, even very aggressive resection, this is frequently impossible due to the lack of accessibility into this anatomic space and the difficulty of vascular interventions, the fear of the general surgeon of hypervascularized, almost spontaneously bleeding tumors, very close to or even invading large vessels [13].. In the presented case, the massive involvement of the abdominal aorta and inferior vena cava were the decisive factor to limit the intervention, although preoperative CT data were inconclusive regarding this aspect. In the territory of retroperitoneal tumors, both primitive and metastases, a multidisciplinary approach is essential: a mixed surgical team, with the presence of a vascular surgeon and appropriate materials for complex vascular interventions, urologists, experts in digestive surgery, but also of radio- and chemo- therapists to apply complementary treatments in selected cases. Not only in retroperitoneal tumors, but also in uterine sarcomas, vascular involvement has been found to be one of the most significant prognostic factors in patient survival [14]. A new staging system has been already considered for uterine sarcomas, stressing on their particular biological behavior [4]. Having this case in mind, the reports of other authors, and the dependence of the treatment on radical surgery, we propose reconsideration in the current approach of uterine tumors. There should be an increase in the level of susceptibility at the presentation of a patient with a polypoid mass, a limitation, as much as possible, of conservative surgery in younger patients, a mandatory extensive histopathological analysis of the entire hysterectomy specimen with routine stromal mitotic count and immunohistochemical determinations, an obligativity of staging lymphadenectomy during the primary surgery (subject of debate until now), a thorough preoperative and intraoperative vascular involvement assessment, a long period of follow-up after the operation, with an attentive screening of the retroperitoneum too and with a careful imagistic vascular observation. This approach might increase, in time, the diagnosis of these tumor variants that probably occur more frequently than reported and would certainly improve the survival of these, often young patients, before reaching, very fast, a very advanced stage.Usually, mullerian adenosarcomas occur after menopause (median age of 58 years), being described from the second to the tenth decade [Research Support: This study has been done in the framework of a FEST project.


Background: Biomonitoring studies are conducted to assess internal dose  to environmental chemicals. However, because of the ubiquitous presence in the environment of some of these chemicals, such as bisphenol A (BPA), external contamination during handling and analysis of the biospecimens collected for biomonitoring evaluations could compromise the reported concentrations of such chemicals.Objectives: We examined the contamination with the target analytes during analysis of biological specimens in biomonitoring laboratories equipped with state-of-the-art analytical instrumentation.Discussions: We present several case studies using the quantitative determination of BPA and other organic chemicals  in human urine, milk, and serum to identify potential contamination sources when the biomarkers measured are ubiquitous environmental contaminants.Conclusions: Contamination with target analytes during biomonitoring analysis could result from solvents and reagents, the experimental apparatus used, the laboratory environment, and/or even the analyst. For biomonotoring data to be valid—even when obtained from high-quality analytical methods and good laboratory practices—the following practices must be followed to identify and track unintended contamination with the target analytes during analysis of the biological specimens: strict quality control measures including use of laboratory blanks; replicate analyses; engineering controls  as needed; and homogeneous matrix-based quality control materials within the expected concentration ranges of the study samples. Humans are exposed to environmental chemicals through industrial and indoor air pollution, diet, and use of personal care and consumer products. Biomonitoring  is widely used to assess human internal exposure  to these chemicals .Proper biomonitoring practices take into account the selection of the relevant biomarker and biomonitoring matrix, the potential impact of the collection protocol on the biomarker levels in the sample, as well as the integrity of the sample during its collection, handling, storage, and analysis . FurtherHowever, even with the application of sophisticated and accurate methods, external contamination with some ubiquitous environmental organic chemicals, such as bisphenol A (BPA), polybrominated diphenyl ethers (PBDEs), and polychlorinated biphenyls (PCBs), during sample analysis can compromise the analytical determination of these compounds . ExternaField blanks have been used to assess potential contamination during collection, processing, and/or transport of environmental samples  and the Several reports have investigated potential sources of contamination with ubiquitous organic chemicals during sample analysis and how to mimimize contamination in a typical biomonitoring laboratory setup . Here weContamination from solvents and reagents. During analysis, contamination from solvents (including water) or reagents is monitored through the calculated concentration and the signal/noise (S/N) ratio of the reagent blank or QCB peak  of the method. Therefore, reagent or solvent contamination can not be revealed by the calculated QCB concentration, but only through the higher than expected S/N ratio of the QCB .QCB peak . However samples . If the To measure BPA in urine, we have used methanol and water as mobile phases of an on-line SPE-HPLC-MS/MS system , 2006. ITo minimize the impact of low-level contamination of HPLC and SPE mobile phases with the target compounds , we modified our experimental configuration by adding guard cartridges to both mobile phase lines, similar to the approach we used before for filtering out an interference of perfluorooctanoate . SpecifiWhen contamination affecting standards, QC materials, and study samples cannot be avoided completely, the concentration of study samples is adjusted by subtracting the contribution from the QCB. However, a high QCB contribution will increase the variability of measurements at low-concentration ranges. In the example above, contamination with BPA of the water purification system increased the frequency of out of control values of the low-concentration QC materials (~ 2.5 µg/L), which, unfortunately, is also around the median urinary concentration of BPA for the general population in various countries . TogetheContamination of experimental apparatus. The experimental apparatus  could be a contamination source when measuring PBDEs , another alternative is including additional QCBs and QCs within the analytical batch.The analyst can also be the contamination source, especially for chemicals present in consumer or personal care products. In another example, we noticed random, yet often repeated contamination with triclosan in some QCBs and QCs included in the batches prepared by one analyst, but not others. After ruling out the potential contamination sources discussed above and confirming that this person followed the required standard operating procedures, we inventoried all of the situations, both in and outside the workplace, where the analyst could come in contact with triclosan-containing products . At the Unintended contamination with ubiquitous environmental chemicals, such as BPA and other organic compounds , during biomonitoring analyses is possible, even with state-of-the-art analytical methods and laboratory facilities. Unfortunately, until all of the environmental sources of these chemicals are known, totally eliminating external contamination is practically impossible. However, judicious application of the measures below will allow the identification of contamination scenarios, thus facilitating the implementation of measures to isolate and track external contamination and minimize as much as possible its recurrence and impact.Evaluate the QCB  by checking both the calculated concentration and the S/N ratio of the QCB peak to assess any systematic or non-systematic contamination.Prepare the QCBs using the same procedure and apparatus as the unknown study samples to avoid missing labware and apparatus potential contamination.Use guard columns to filter out potential interference contaminants from the SPE mobile phases or separate the chromatograhic peak of the target analyte in the study samples from its interference peak in the HPLC mobile phases.Conduct replicate analyses  of study samples to identify random contamination from the laboratory environment and/or analyst, especially when the number of study samples is small. When limited sample volume precludes replicate analyses, include additional blanks randomly placed within the analytical batch.Prepare the samples for analysis in a controlled environment  if contamination from the analyst or the environment  is suspected.Use homogeneous matrix-matched QC materials at concentrations within the expected concentration ranges of the study samples.When possible, participate in external quality assessment programs or use SRMs to evaluate the accuracy of the measurement. Unfortunately, for nonpersistent environmental organic chemicals  commercially available SRMs do not exist and only BPA is routinely included in G-EQUAS. Expanding such quality assessment programs and characterizing SRMs to include these chemicals are critical to improve the accuracy of biomonitoring methods.In summary, valid biomonotoring data require integrating numerous preanalytical and analytical steps. First, adequate selection of the most relevant biomarkers and biomonotoring matrices as well as collection, handling, shipping, and storage procedures to preserve the integrity of the specimen and the target analytes are needed before starting sample analyses. Second, use of sensitive, selective, and accurate analytical methods and state-of-the-art laboratory facilities. Third, use of highly trained laboratory personnel not only to operate sophisticated instrumentation, but also to recognize situations that may compromise both the integrity of the biological specimen and the validity of its analysis. Last, use of good laboratory practices and implementation of measures as discussed above to minimize unintended contamination of the biological specimens with the target analytes during analysis. We strongly believe that biomonitoring studies that maximize the harmonization of the various disciplines and expertises  will considerably strengthen the exposure assessment.


Populus are particularly noted for their drought sensitivity; therefore, understanding the mechanisms by which these economically and ecologically important forest trees respond to drought is of paramount importance. The ability of Populus trees to contend with drought is dependent on the responsiveness of the genome, and in turn, the ability of the transcriptome to appropriately remodel growth, metabolism and development. Amassing evidence indicates that different species of Populus have divergent mechanisms and adaptations to contend with drought stress; however, individuals within a given species also display divergent drought responses. In order to investigate the intra-specific variation underpinning the drought response, we examined six genotypes of P. balsamifera. Using Affymetrix Poplar GeneChips, we found a positive correlation between the magnitude of drought-induced changes in the transcriptome and the capacity of the genotype to maintain growth. Surprisingly identifiable differences at the transcriptome were observed, and similar responses were observed within the metabolome. Although common drought responses could be identified within the species, the complexities of these responses must be taken into consideration when defining species- or genus-level drought responses.Drought is one of the most significant factors limiting tree growth. Trees in the genus


A 43-year-old man from a rural area in Brazil presented to his physician with complaints of palpitations and lightheadedness, mainly on exertion. An electrocardiogram (ECG) showed right bundle-branch block (RBBB), left anterior fascicular block (LAFB), and a premature ventricular contraction (PVC) . EchocarChagas heart disease (chronic chagasic cardiomyopathy). Chagas heart disease (CHD) is the most common etiology of cardiomyopathy in Latin America, and a major cause of cardiovascular death among middle-aged individuals in endemic areas. CHD is also the most serious and frequent manifestation of chronic Chagas disease. It develops in 20%–30% of infected persons, usually 10–30 years after Trypanosoma cruzi contamination, and manifests as three major syndromes that often coexist in the same patient: arrhythmias, heart failure, and thromboembolism Arrhythmias are very common, of different types, and may cause palpitations, dizziness, syncope, and sudden cardiac death (SCD). Frequent, complex PVCs, including couplets and runs of NSVT, are a common finding on 24-h Holter monitoring and exercise testing. They correlate with the severity of ventricular dysfunction, but can also occur in patients with quite well preserved ventricular function. Episodes of NSVT are seen in approximately 40% of patients with mild wall motion abnormalities and in virtually all patients with heart failure, an incidence that is higher than that observed in other cardiomyopathies Heart failure is often a late manifestation of CHD. It is usually biventricular with a predominance of left-sided failure at initial stages and of right-sided failure at more advanced disease. Systemic and pulmonary embolisms arising from mural thrombi in the cardiac chambers are quite frequent. Clinically, the brain is by far the most frequently recognized site of embolisms (followed by limbs and lungs), but at necropsy, embolisms are found more frequently in the lungs, kidneys, and spleen Electrocardiographic alterations are varied, but the most common are RBBB, LAFB, PVCs, primary ST-T changes, Q waves, low QRS voltage, different degrees of atrioventricular block, and manifestations of sinus node dysfunction. All these alterations may be isolated or associated. The association of RBBB with LAFB strongly suggests chronic CHD in endemic regions On the chest radiography, cardiac size is normal in the initial stages of disease and even when multiple ECG abnormalities are present. The cardiac size may be slightly, moderately, or severely increased at later stages of CHD. In nearly half of the cases with heart failure, the manifestations of pulmonary congestion are poor or even absent Holter monitoring is an excellent method for investigating patients with CHD. It may be used to identify complex ventricular arrhythmias, diagnose transitory arrhythmias, and evaluate anti-arrhythmic therapy. The exercise testing evaluates the functional capacity of the patient, qualifies and quantifies PVCs, and may verify the efficacy of anti-arrhythmic drugs.T. cruzi antigens needs to be detected by at least two different serological methods  to confirm the etiological diagnosis.In patients with high suspicion of chronic Chagas disease or in those with a compatible clinical syndrome, because parasitaemia is scarce, the presence of immunoglobulin G (IgG) antibodies against T. cruzi infection includes a complete medical history and physical examination, and a resting ECG The initial evaluation of the newly diagnosed patient with chronic Recently, some of us used a rigorous multivariate analysis to develop a risk score for mortality prediction in 424 outpatients with CHD from a regional Brazilian cohort The clinical history, the ECG abnormalities, and the findings on echo, Holter, and exercise testing in the present case are characteristic of chronic CHD. Serological tests for Chagas disease were performed and gave positive results. According to the scoring system to determine the risk of death described above, this patient would receive a score of 8 points based on his sex (2 points), the presence of apical aneurysm (3 points), and the presence of runs of NSVT (3 points). This score puts him in the intermediate-risk category (risk of death in the next 10 years of 44%).The importance of having the results of a chest radiography on board, which was not done in this case, relies on the fact that the presence of cardiomegaly would add 5 more points to the patient's risk score, moving him to a higher-risk category. Of note, the combination of NSVT and LV systolic dysfunction in patients with CHD increase the risk of subsequent death by 15 times compared to patients without these abnormalities on Holter monitoring and on echocardiography Management of patients with CHD should focus on eradication of the parasite, which may halt disease progression, alleviation of symptoms of disease, and identification of patients at high risk of death, in order to implement strategies with the potential to reduce mortality. Regarding antitrypanosomal treatment, only two agents are currently available: benznidazole and nifurtimox. On the basis of the results of some observational studies, a panel of experts convened by the US Centers for Disease Control and Prevention in 2006 recommended that antiparasitic therapy should generally be offered to adults 19–50 years of age with mild to moderate cardiomyopathy Because some evidence suggests that mild segmental LV wall motion abnormalities are predictors of deterioration of ventricular function during follow-up The ethics committee of Anis Rassi Hospital approved this report and waived the need for informed consent.Clinical manifestations of Chagas heart disease are diverse and related to three major syndromes: cardiac arrhythmias, heart failure, and thromboembolism. Improved models for risk stratification are now available, and certain guided treatments may increase survival.Segmental or global left ventricular dysfunction and episodes of nonsustained ventricular tachycardia are independent, additive risk factors for overall mortality in Chagas cardiomyopathy.Although not supported by incontestable evidence, it is possible that patients with runs of nonsustained ventricular tachycardia on Holter/exercise testing, and asymptomatic or mildly symptomatic left ventricular dysfunction on echocardiography, derive some benefit from combined therapy with an angiotensin-converting enzyme inhibitor and amiodarone.T. cruzi infections or even with mild to moderate disease.Persistence of parasites and the accompanying chronic inflammation is the basis for the pathology in Chagas heart disease. Thus, unless proved otherwise by a randomized controlled trial, benznidazole or nifurtimox should usually be offered to patients <50 years old with presumably long-standing indeterminate The potential benefits of novel therapies  need assessment in prospective randomized trials. The best therapeutic strategy for patients with apical aneurysm not containing thrombus is unknown.


Consistent evidence links major depression and its affective components to negative health outcomes. Although the pathways of these effects are likely complex and multifactorial, recent evidence suggests that innate inflammatory processes may play a role. An overview of current literature suggests that pathways between negative moods and inflammation are bi-directional. Indeed, negative moods activate peripheral physiologic mechanisms that result in an up regulation of systemic levels of inflammation. Conversely, peripheral inflammatory mediators signal the brain to affect behavioral, affective and cognitive changes that are consistent with symptoms of major depressive disorder. It is likely that these pathways are part of a complex feedback loop that involves the nervous, endocrine, and immune systems and plays a role in the modulation of peripheral inflammatory responses to central and peripheral stimuli, in central responses to peripheral immune activation and in the maintenance of homeostatic balance. Further research is warranted to fully understand the role of central processes in this feedback loop, which likely contributes to the pathophysiology of mental and physical health. Evidence shows an association of major depression with increased risk for adverse physical health outcomes. Indeed, depression, whether assessed as a continuum of symptoms or as the presence of a clinical syndrome, predicts the incidence and progression of diseases of aging, including cardiovascular, metabolic and neurodegenerative diseases, as well as all-cause mortality -3. GivenEarly studies show an association of depression with the down-regulation of functional parameters of the immune system . HoweveThe inflammatory response is a non-specific immune reaction that is initiated when monocytes/macrophages are activated by pathogens or tissue damage to release pro-inflammatory cytokines, such as interleukin (IL)-6, IL-1β, and tumor necrosis factor (TNF)-α. These cytokines initiate a local and systemic inflammatory response, which includes the hepatic synthesis and release of acute phase proteins, such as C-reactive protein (CRP) and fibrinogen . PeripheCirculating levels of pro-inflammatory mediators are widely accepted as a marker of systemic levels of inflammation. However, caution should be taken in assuming these inflammatory mediators are immune-derived as many cells produce these signaling proteins, including adipocytes and endothelial cells . RegardlAvailable data suggest that relationships between pro-inflammatory cytokines and depressed mood are bi-directional. In support of immune-to-brain pathways, sickness symptoms mediated by increases in circulating pro-inflammatory cytokines are consistent with symptoms of depression including fatigue, sleep disturbances, anxiety, negative mood, anhedonia, and loss of appetite . Indeed,To examine the impact of peripheral inflammation on the central nervous system, recent attention has focused on whether immune-related patterns of brain activation are consistent with those that accompany clinical depression. Here, animal studies show that pro-inflammatory cytokines can penetrate the blood-brain barrier to stimulate the production of central pro-inflammatory cytokines by microglial cells in discrete brain regions that are involved in mood regulation and reward processing . Recent In addition to immune-to-brain pathways, evidence also shows that negative mood states, stressful experiences, and antagonistic dispositions can activate peripheral physiologic pathways that modulate immune function. For example, negative moods are associated with activation of the hypothalamic-pituitary-adrenal (HPA) axis and the peripheral release of cortisol, along with increased activation of the sympathetic and decreased activation of the parasympathetic branches of the autonomic nervous system ,34. Theset al. [In sum, converging evidence supports reciprocal pathways linking inflammation and the disruption of mood. It is likely that these pathways are part of a complex feedback loop that involves the neuroendocrine and immune systems and plays a role in both the modulation of peripheral inflammatory responses to stimuli and the maintenance of homeostatic balance. Further research examining specific symptoms and the central processes involved in these circuits is warranted to fully understand the role of inflammation in the pathophysiology of depression and of associated health morbidity. Evidence suggests that examining the impact of peripheral inflammation on central processes that play a role in specific symptoms of depression may point to novel targets for future intervention. For instance, Motivala et al.  found thThe authors declare that they have no competing interests.BM conducted an up to date review of the relevant literature and wrote the initial draft of the manuscript. ALM edited the manuscript and made substantial revisions. AL read, contributed relevant research, and provided feedback on early and final versions of the manuscript. All authors read and approved the final manuscript.ALM: Anna Marsland, Ph.D. is the Director of the Behavioral Immunology Laboratory and a member of the Biological and Health Psychology faculty at the University of Pittsburgh. Her expertise is in the field of psychoneuroimmunology. BM: Berhane Messay is a second year graduate student in the Clinical and Biological/Health Psychology Programs at the University of Pittsburgh. AL: Alvin Lim is a second year graduate student in the Biological/Health Psychology Program at the University of Pittsburgh.


Iron deficiency is still the world's most common nutritional deficiency, and generally, iron deficiency anemia is the most prevalent form of anemia. The major risk groups for iron deficiency include women of childbearing age, pregnant women, and lactating postpartum women. There exist plenty of studies attending and reconfirming this important issue. To combat this problem, different food iron fortification programmes have been implemented in some regions, iron supplementation guidelines have been elaborated in some countries, and iron therapy schemes involving both oral and intravenous administration have been proposed. But despite these efforts and the recommendations by the World Health Organization, the problem concerning iron deficiency and anemia is still unsolved in most parts of the world. Even in many industrialized countries, the topic of iron and pregnancy often stays unattended and there is a lack of consensus concerning guidelines in different countries.Other disorders, such as hemoglobinopathies, also contribute to the high prevalence of anemia worldwide. The prevalence of anemia is influenced by a variety of deficiencies , infectious diseases, parasitic infestations, inflammatory diseases as well as malnutrition. As data for iron are quite profound, these are often scarce if we look on other vitamins and micronutrients, except for folic acid. As we know, periconceptional supplementation with folic acid has the potential to reduce the incidence of neural tube defects  dramatically. Unfortunately, folic acid prophylaxis is not commonly used for several reasons.By this special issue, our intention was to broaden the perspective on anemia in pregnancy and propose ways to solve the problems. We have chosen three articles concerning the prevention/treatment of iron deficiency anemia in pregnancy/postpartum including papers from Australia, Nigeria, and Denmark. Another Danish study focuses on the prevention of postpartum anemia through hemostatic sealing of the placental bed at caesarean section, thereby reducing perioperative blood losses. A paper from France and Lebanon evaluates the relationship between periconceptional folate deficiency and neural tube defects. A Chinese study addresses the supposed relationship between iron deficiency and postpartum depression. Together, these papers illustrate some of the many facets of the complexity of the prevention and treatment of anemia in pregnant and postpartum women.Alexander KrafftAlexander KrafftLaura Murray-KolbLaura Murray-KolbNils MilmanNils Milman


P < 0.001).Alzheimer's disease (AD) is a progressive neurodegenerative disorder characterized by a gradual decline in memory associated with shrinkage of brain tissue, with localized loss of neurons mainly in the hippocampus and basal forebrain, with diminished level of central cholinergic neurotransmitter-acetylcholine and also reported to be associated with accumulation of ubiquitinated proteins in neuronal inclusions and also with signs of inflammation. In these disorders, the abnormal protein aggregates may themselves trigger the expression of inflammatory mediators, such as cyclooxygenase 2 (COX-2). In the present study, the effects of Meloxicam, Selegiline, and coadministration of these drugs on scopolamine-induced learning and memory impairments in mice were investigated. Rectangular maze test, Morris water maze test, Locomotor activity, and Pole climbing test were conducted to evaluate the learning and memory parameters. Various biochemical parameters such as acetylcholinesterase(AChE), TBARS assay, catalase activity, and DPPH assay were also assessed. The present study demonstrates that Meloxicam, Selegiline, and co-administration of these test drugs had potential therapeutic effects on improving the antiamnesic activity in mice through inhibiting lipid peroxidation, augmenting endogenous antioxidant enzymes, and decreasing acetylcholinesterase activity in brain. The memory enhancing capacity of the drugs was very significant when compared to disease control ( Loss of cholinergic cells, particularly in the basal forebrain, is accompanied by loss of the neurotransmitter acetylcholine . Increased MDA level as one of the ROS has been shown to be an important marker for in vivo lipid peroxidation. Recently, many studies reported that memory impairment in the scopolamine-induced animal model is associated with increased oxidative stress within the brain , 30, 31. From the behavioral test, that is, rectangular maze test and Morris water maze test, it is clearly seen that there was a general decrease in the transfer latency in all treated groups compared to the scopolamine-treated group. The memory loss effect of scopolamine is more prominent compared to the control group. In comparison with Donepezil, the drug-treated groups had almost equal performance which indicates synergistic effect of meloxicam and selegiline against memory loss. Meanwhile locomotor activity and pole climbing avoidance tests are done which also indicate the leaning ability . The major antioxidant and oxidative free radical scavenging enzymes like glutathione, SOD, and catalase play an important role to reduce oxidative stress in brain. In this study, from the DPPH assay antioxidant levels are estimated. These enzyme levels are decreased in the scopolamine-treated group compared to the control group. The enzyme levels are almost equal in combination group and the standard group. Individual groups are showing less than standard group. It supports the antioxidant action of drugs. In the present study rats after scopolamine treatment showed a significant increase in the brain levels of malondialdehyde, which is the measure of lipid peroxidation and free radical generation. In the drug-treated groups, there is a significant decrease in the levels of malondialdehyde which is nearly equal to the standard group. From the results, it is clear that the anti-inflammatory activity of meloxicam decreases the disease progression. The antioxidant activity of selegiline is clear from the biochemical tests, which includes the estimation of antioxidant enzymes. In conclusion, the present study demonstrates that Meloxicam, Selegiline, and co-administration of these test drugs had potential therapeutic effects on improving the antiamnesic activity in mice through inhibiting lipid peroxidation, augmenting endogenous antioxidant enzymes, and decreasing acetylcholinesterase (AChE) activity in brain.