The use of fluorescent nanoparticles as probes for bioanalytical applications is a highly promising technique because fluorescence-based techniques are very sensitive. Quantum dots (QDs) seem to show the greatest promise as labels for tagging and imaging in biological systems owing to their impressive photostability, which allow long-term observations of biomolecules. The usage of QDs in practical applications has started only recently, therefore, the research on QDs is extremely important in order to provide safe and effective biosensing materials for medicine. This review reports on the recent methods for the preparation of quantum dots, their physical and chemical properties, surface modification as well as on some interesting examples of their experimental use. The reaa is lower than one of this magnitudes: ae, ah and aexc . It is known the Bohr radius depends on the material  and therefore there is not a clear line to say that a nanoparticle is a QD or not, if one only considers its size. QDs can be based on metallic  [Nanometer-sized crystals, often referred to as quantum dots (QDs), have been also intensively investigated. Typical QD sizes range between 2–20 nm , but acc Pt, Au) . Confine Pt, Au) . In addi Pt, Au) ,11.2.2.1.in vivo QDs application that cadmium ions were determined as the primary cause of cytotoxicity, because they are able to bind to thiol groups on critical molecules in the mitochondria and cause enough stress and damage to cause significant cell death [2+ could be reduced or eliminated by adding additional surface coatings. Scientists in Ireland have been using gelatin during the production of CdTe QDs, thereby reducing the toxicity of the particles [et al. used as stabilizer the tripeptide glutathione, which exists in most organisms and thus can be applied to detoxify Cd2+ ions in Medicine due to its chelating capability [et al. reported that peptides could be used as an excellent ligand for coating the surface of QDs for several reasons: 1) peptides mimic the biological environment and are stable at physiological pH; 2) reactive groups  can be dialed into the hydrophilic domain of the peptide sequence and enable enzymatic or standard conjugation chemistry to obtain biomolecules of interest; 3) molecular evolution strategies to randomize peptides can be adapted to select high-affinity binders to the QD surface; and 4) there is the possibility of creating multifunctional QDs by mixing peptide sequences in certain molar ratios in a single step for in vitro and in vivo studies  [Unfortunately, most QDs are toxic, which implies a potential danger, especially for future medical applications. The most widely used and studied QDs consist of a core of cadmium selenide or telluride, because their quantum confinement region spans the entire optical spectrum . Wallingll death . Howeverarticles . In othepability . Iyer etG, etc.) .In other words, surface functionalization plays the key role in nanoparticle toxicity. It was found the improved biocompatibility that certain surface coatings provide could arise from decreased cellular uptake of these nanoparticles for cellular degradation . Accordiin vivo toxicity of QDs with carboxyl and amine surface coatings during investigation of the inflammatory properties, tissue distribution, and prothrombotic effects after intravenous injection into mice [in vivo imaging experiments. Nevertheless, a key question is still open: whether QDs can be used directly in clinical phase studies? Consequently integrated cellular toxicity studies and the complete in vivo toxicology of QDs still need to be evaluated before their use in human applications.According to many studies, short-term (acute) and long-term toxicity of QDs is an important issue for QD applications in bio systems; not only at the cellular level, but also in animal models –22. Cellnto mice . Yong dento mice . Therefonto mice . They foThe other important criteria for QDs using in medicine are prevention of nanoparticle aggregation in a biological environment and effective suppression of non-specific adsorption of biomolecules at the nanoparticle surface. The insufficient colloidal stability can be improved by functionalization of QDs surface with hydrophilic polymers such as PEG  or with In contrast to organic fluorophores, QDs have some unique photophysical properties. They have a continuous absorption spectrum for wavelengths shorter than the wavelength of fluorescence emission. QDs can be synthesized to be effectively monodispersed which is a cause that their emission spectra are quite narrow and symmetric, and do not show any red-tail. In this way many different colors can be excited with just one wavelength of excitation and can be spectrally well resolved . For exaHowever there are two disadvantages in comparison to classic fluorescent dyes. The first negative point of QDs is the lack of strong polarization of the emitted photons, mainly in the case of elongated QDs, also called nanorods . The secet al. also found the photoluminescence lifetime of intracellular thiol-capped CdTe QDs varied greatly depending on the intracellular environment: in the acidic lysosomes (with a pH of around 4.5–5.0) was remarkably shorter than those in other parts of the cell [As it is known, the photoactivation of the semiconductor nanoparticles is highly affected by several parameters. Dembski and colleagues have systematically investigated the role of those parameters on the photon-induced photoluminescence enhancement of CdSe/ZnS QDs embedded in silica colloids (multicore particles) and polymer-stabilized CdSe/ZnS QDs in various environments . The stuthe cell . On the the cell .2.2.Concerning the biological applications of QDs, two main groups may be cited: biosensors and labels in biological imaging. A few examples of each group can be seen on the schema below .QDs are excellent candidates for biosensing owing to their unique physical and optical properties and possibility of attaching various biomolecules to their surface . Some neMost biomolecules have been linked to water-soluble QDs and it was found the binding has no effect either on the QDs’ optical signature or on the functionality of the biomolecules. Biomolecules can be bound to the surface of QDs either directly  or attached via a stabilizing layer which act as a crosslinker between the ligand and reactive surface of the nanoparticle. Non-covalent direct binding can be achieved by applying of electrostatic-based coupling strategy. Compared to the conventional covalent techniques, the electrostatic non-covalent self-assembly approach is simpler, more reproducible, and more easily achieved . This apN-hydroxysulfosuccinimide (NHS) to form the QD-protein covalent conjugates is one of the most frequent coupling methods. The research group of Wang determined that 3-mercaptopropyl acid-stabilized water-soluble CdTe nanoparticles synthesized in aqueous solution were able to conjugate with peptides or proteins mediated by NHS [The covalent bioconjugation approach is based on the replacement of thiol acids present on the QDs’ surface with thiolated biomolecules. This type of binding was chosen, for example, to covalently link streptavidin maleimide  or to cod by NHS . These sIn these applications, core-shell structured QDs are more favorable, where CdSe is referred to a core and ZnS to a shell –47. The in vitro and in vivo. Scientists have developed hybrid functionalized QDs-liposome nanoparticles and found they were efficiently uptaken by living cells in the absence of cell death and can therefore be used as fluorescent probes for ex vivo cell-labeling studies with most types of water-soluble QD without further modifications [in vitro (tumor spheroids) and in vivo (subcutaneous solid tumors). Pan and colleagues evaluated in vitro QDs loaded in poly(lactide)-vitamin E TPGS (d-α-tocopheryl polyethylene glycol 1000 succinate) nanoparticles for cellular and molecular imaging [et al. studied an application of QD-labeled biomolecules in a sandwich-type immunoassay for detection of total prostate-specific antigen (TPSA), which is an important cancer marker, on a screen-printed carbon substrate in connection with fluorescence imaging [There are several possibilities for using QDs in biolabeling and cellular imaging both ications . Moreove imaging . Kerman  imaging . Similar imaging .in vivo, mainly inside small animals, where they can substitute conventional organic fluorophores emitting in the IR, which suffer from poor stability and quantum yield [et al. evaluated in vivo multiplex imaging of mouse embryonic stem (ES) cells labeled with peptide-based Qtracker delivered QDs [QDs can be applied in molecule tracking in immunochemistry, where they replace the fluorescent beads used for the study of the dynamics of neurotransmitter receptors. Due to their much smaller size (about 10–20 nm) compared to latex beads (approx. 500 nm), the lateral movement of individual receptor can be studied in great detail . Anotherum yield , 60, 61.ered QDs   formation of nanosized semiconductor particles through colloidal chemistry ,68 and  or liquid phase epitaxy (LPE). Although LPE is still important, this technique was used mostly in past century to prepare micrometric structures. VPE can be further categorized according to reaction type into chemical (CVD) or physical vapor deposition (PVD). Chemical deposition is interesting in view of its price, however the most important and most widely used technique in industry is VPE from metalorganic medium (MOVPE). Molecular beam epitaxy (MBE) is other peculiar PVD technique and can be also classified into solid source MBE or gas source MBE . Perhaps less well-known approach for the growth of nanostructures using MBE system is “droplet epitaxy” [Epitaxial method of QDs preparation is widely used in optoelectronics  and nanotechnologies. However, in the near future, it could be very interesting and promising to use epitaxially grown QDs in epitaxy” . Here liConcerning the QDs direct patterning, self-assembled monolayers QDs can be coated via Langmuir-Blodgett technique onto hydrophobic–hydrophilic pattern generated with employing of photolitographically prepared template .et al. prepared metal nanodots using the electrochemical deposition through cavities formed within macroporous poly(pyrrole) secondary templates which were themselves prepared by electrodeposition around templates formed of self-assembled poly(styrene) spheres assembled on evaporated Au substrates [2 nanodots with controllable size on a Si substrate, where an alumina filtration membrane with homogenous micropores acts as the “microscopic mass-point” nozzle [4F with ethylene glycol [For the past few years, template-based nanoengineering methods, in which many different materials such as porous alumina, polymer gel, surfactant, activated carbon and carbon fiber have been used as templates to synthesize different kinds of nanostructured porous materials, have been extensively studied. For example Ghanem bstrates . Recentl” nozzle . The tit” nozzle . During e glycol .2 nanodots is very much different from that of TiO2 thin films and powders. After high temperature annealing, the nanodots are polycrystalline and consist of a mixed phase of anatase and rutile instead of single rutile phase. Scientists expect that TiO2 nanodots with a single phase of anatase can be prepared from an epitaxial Al/TiN bilayered film on a sapphire substrate by electrochemical anodization of the TiN layer using a nanoporous anodic aluminum oxide film as the template [2 QDs properties is very complex, because several parameters affect the final quantum confinement (the multiple influence of QDs size and coordination of Ti atoms on Ti–O bond length and thus on band gap energy) [The phase development of the isolated TiOtemplate . The ana energy) ,92.in vitro applications. The enhanced photocatalytic effect of TiO2 nanoparticles in skin and other types of cancers was recently demonstrated [2 nanoparticles was shown to have a unique property of a light-inducible nucleic acid endonuclease, which could become a new tool for gene therapy. The preparation of TiO2 nanodots with small dot size and high dot density on substrates becomes increasingly significant since small dot size can guarantee quantum effects and high activity, while high dot density can provide a basis for miniaturizing devices and increasing performance capacity [2 on the electrode could enhance the enzyme catalytic performance for promising biosensor applications. However, the electrochemical behavior of TiO2 depends not only on the crystal structure and surface properties, but also on the textural properties, which include specific surface area, pore volume, pore dimension and distribution. Great effort has been dedicated to the synthesis of TiO2 with a controllable morphology and pore structure [2 QDs electronic structure via the first-principles band structure method [The use of non-toxic titanium dioxide can completely suppress the problem of QDs toxicity in biomedicine for capacity . It has tructure . Scientie method .3.2.Optical characterization of QDs is usually provided by UV-VIS and photoluminescence spectroscopy, which offer fast, non destructive and contactless option. Hazdra and colleagues employed a rarely used experimental method, photomodulated reflectance spectroscopy, for checking deposited QD structures. This technique provides equivalent energy resolution to that of photoluminescence at low temperatures and probes a wider range of critical points, highlighting the ground state and also many high order interband optical transitions from which band structure, especially of the wetting layer, can be deduced .As already mentioned, the optical properties (fluorescence emission) of QDs can be fine-tuned by the QDs’ size, which is a key parameter that determines the spectral position and purity of photoluminescence. QDs’ size is generally calculated using conventional techniques like scanning electron microscopy (SEM), transmission electron microscopy (TEM), and dynamic light scattering (DLS) studies. Gu and colleagues determined the size and composition of optically active CdZnSe/ZnBeSe QDs using photoluminescence, photoluminescence excitation, and Raman scattering spectroscopies combined with a model of photoluminescence and LO phonon energies . BesidesIn order to monitor the size of epitaxially prepared QDs, several characterization methods are usually employed, e.g. TEM, atomic force microscopy (AFM) or more preferably scanning tunneling microscopy (STM) as can be seen from et al., the QDs’ surface chemistry plays also very important role, because it contributes to the final hydrodynamic diameter of QDs and thus it is necessary to find a method providing qualitative information about this parameter [et al. characterized the surface chemistry and size distribution of ODs using analytical ultracentrifugation method, which was found to be highly sensitive to nanocrystal size, resolving nanocrystal sizes that differ by a single lattice plane.According to Lees arameter . To date4.in vivo. Therefore scientists are making great efforts to solve this problem, generally by coating QDs surface with protecting and stabilizing shell, in order to use them safely for subsequent bioconjugation with proteins, peptides or other chemical moieties. It was also found that round shape of QDs is preferred when using them in solution for biosensing, so called colloidal QDs, as it was referred in majority of papers, while the deposited QDs with shape of islands are generally required for optoelectronic media.QDs have been found useful due to their sensing capability for various particular applications, in fields such as optoelectronics and medicine. The main reasons for their use for medical purposes are the following: excitation wavelength far from the emission, inherent photostability and long fluorescent lifetime. It is evident that an appropriate material for biosensing devices will be very different from the one suitable for optical devices. In particular the question of QDs’ toxicity in the former applications is very important, mainly for essays performed Concerning the medical applications, there are still several issues, like desirable orientation of attached biomolecules, which remain to be resolved, so it can appear that commercial QDs are more appropriate for some approaches. Hence, it is necessary to continue the QDs research to develop their surface functionalization and conjugation with biomolecules more precisely. From biomolecules detection point of view, it is also interesting to study the behavior of QDs immobilized on variety of supports.


Drug resistance is generally considered to be a major impediment to successful cancer chemotherapy, yet it is generally not possible to predict the degree or timing of the emergence of tumour resistance in most chemotherapy protocols. Recent developments with the single-cell gel electrophoresis or 'comet' assay for DNA damage at the single-cell level suggest that this technique might provide a method for identifying and potentially monitoring tumour cell responsiveness to many anti-cancer agents in situ. In principle, this assay could be applied to any accessible tumour being treated with chemotherapeutic agents that cause overt DNA damage. We have investigated that supposition using several rodent and human tumour cell lines exhibiting a spectrum of resistance to the DNA strand-breaking drug, etoposide. By assessing cells grown as monolayers, spheroids and xenografted tumours in immunodeficient mice, we found that the comet assay can provide not only an index of sensitivity to etoposide, but, additionally, can demonstrate the efficacy (or lack thereof) of multidrug resistance (MDR) reversing agents for cells in vitro, and tumours in vivo.


Recent years have seen a huge increase in the amount of biomedical informationthat is available in electronic format. Consequently, for biomedical researcherswishing to relate their experimental results to relevant data lurking somewhere withinthis expanding universe of on-line information, the ability to access and navigatebiomedical information sources in an efficient manner has become increasinglyimportant. Natural language and text processing techniques can facilitate this taskby making the information contained in textual resources such as MEDLINEmore readily accessible and amenable to computational processing. Names ofbiological entities such as genes and proteins provide critical links between differentbiomedical information sources and researchers' experimental data. Therefore,automatic identification and classification of these terms in text is an essentialcapability of any natural language processing system aimed at managing the wealthof biomedical information that is available electronically. To support term recognitionin the biomedical domain, we have developed Termino, a large-scale terminologicalresource for text processing applications, which has two main components: first, adatabase into which very large numbers of terms can be loaded from resources suchas UMLS, and stored together with various kinds of relevant information; second,a finite state recognizer, for fast and efficient identification and mark-up of termswithin text. Since many biomedical applications require this functionality, we havemade Termino available to the community as a web service, which allows for itsintegration into larger applications as a remotely located component, accessed througha standardized interface over the web.


Objective: Sex reassignment surgery is accepted in properly planned cases to change facial appearance in accordance with a new gender. Although changing the hairdo or makeup, and depilation, give satisfying cosmetic results in many patients, hard and soft tissue facial surgeries are needed in most cases. Unfortunately, few studies on facial corrections have been published and feminizing a male face is still considered an arbitrary undertaking. We present a useful surgical protocol to feminize the male face. Method: Ten male-to-female transsexuals aged 20 to 32 years (average 23 years) were referred after gender reassigning for facial feminization (1990–2007). Clinical examination of the patients revealed variable degrees of common masculine features such as long square faces, broad chins, excessive projection, frontal bossing, prominent flaring gonial angles, high hairlines, and low-set eyebrows. After complete clinical assessment and paraclinical workup including cephalometry, the patients were admitted for feminizing surgery. The face was outlined in geometric proportions. It was aimed to round the male face using a sequence of staged surgical procedures for both hard and soft tissues. Results: Ten male patients aged 20 to 32 years (average 23 years) underwent facial feminization using basic surgical guidelines. Various amounts of soft and hard tissue changes were required for individual patients and were tailored for each case according to the preoperative workup and standard parameters. Conclusion: All 10 patients were satisfied with their improvement in appearance; the degree of impact these procedures may have on lifestyle remains to be assessed. The sex of a person is what one sees while gender is what one feels.Ten male-to-female transsexuals aged 20 to 32 years (average 23 years) were referred for gender-reassigning facial surgery and feminization from 1990 to 2007. Clinical examination in these patients revealed variable degrees of common masculine features such as long square faces and broad chins with excessive projection, frontal bossing, prominent flaring gonial angles with high hairlines and low-set eyebrows Figs .Our experience in treating 10 male-to-female transsexuals aged 20 to 32 years (average 23 years) correlated a set of effective, general, and surgical soft and hard tissue procedures and guidelines for facial feminization. Considerable feminization was achieved after these procedures in all our subjects. Various amounts of soft and hard tissue changes were required for individual patients depending on facial features and findings. Thus, all these procedures may not be required and also should be tailored for each individual case according to the preoperative workup and assessment and comparison with standard parameters  was issued in this regard. Since then, the incidence of this procedure has been on the increase and does not relate to politics or the political atmosphere whatsoever; rather, it is a therapeutic issue. Religious decrees concerning therapeutic policies are issued via high-ranking religious authorities (Mujtahids) as are social issues. As science advances, decrees may be issued or modified to conform and adapt religious doctrines to the present era and to manage emerging problems in contemporary society. Today, SRS, when deemed necessary by the patient's doctor, has become accepted in our country as well as many other countries in properly planned cases to suit the new gender. Doctors are authorized to carry out such procedures when deemed necessary by the treatment team and when this is envisioned to be in the best interests of the patient.Sex reassignment surgery (SRS) was uncommon in our country before 1990 because this topic was never put to debate and there was no decree on the subject. However, in 1991 this issue was put to debate by the religious clergy and was decreed to be a permitted therapeutic procedure by the highest religious authority; a A combination of surgical procedures is required for male feminization to alter the masculine hard and soft tissue features. Application of these surgical techniques can produce favorable results. The available data have shown that in properly selected patients, surgery yields significant improvements in lifestyle, social relationships, self-esteem, body image, employment status, and sexual adjustment. Most investigators describe the outcome of SRS as satisfactory.


Wayne Hall and colleagues discuss the limitations of genomic risk prediction for population-level preventive health care. Before genomic information is used in public health screening, it must be shown that:such information predicts disease risk better than phenotypic information;cost-effective interventions exist for those at increased genetic risk;these interventions are more cost-effective than population-level interventions;genetic risk information motivates desired behaviour change.Currently there are no examples of genetic screening for disease risk that satisfy these criteria.In the 1990s, during the era of the Human Genome Project, many researchers were very optimistic about the capacity of such large-scale genetic projects to revolutionize the prevention of disease e.g., . Many prTen years on, genome-wide association studies (GWAS) have changed our understanding of the aetiology of many common diseases such as type 1 diabetes and obesity, but they have not identified major susceptibility alleles for most common diseases. With a few exceptions, susceptibility alleles for the most common human diseases have proven to be very weakly predictive of disease risk, with odds ratios for individual alleles typically ranging from 1.1 to 1.6 Despite these limitations, many researchers continue to advocate the use of genetic information to predict disease risk e.g.,  and a nuIn principle, individual genetic variants could potentially provide reasonable prediction of disease risk, if the findings for multiple susceptibility alleles were combined statistically In many contexts, information from multiple genetic variants does not appear to provide better prediction than known risk factors such as family history or environmental risks. For instance, Lango and colleagues The difference in success of prediction between these empirical studies is perhaps not surprising; disease prevalence and heritability are important determinants of the clinical utility of a genetic test. Also, genetic associations often vary by population, because population level variations in location, ethnicity, age, and other factors influence the prevalence not only of genetic risk factors for common diseases, but also of environmental and behavioural risk factors for common diseases, such as tobacco and alcohol use, diet, and exercise Advocates of the preventive use of genetic risk information often simply assume that preventive interventions will be cost-effective if genotypic information can be shown to predict disease risk e.g. . From a However, even if efficacious interventions are available, we need large controlled trials to assess whether providing these interventions to asymptomatic individuals at increased genetic risk is more cost-effective than treating all persons displaying physiological risk factors (such as elevated blood pressure or cholesterol) Some advocates of genomic medicine simply assume that giving genetic risk information will prompt individuals to change their behaviour in desired directions There is some evidence that genetic risk information may make individuals assume that prevention requires pharmacological intervention It is also unclear whether genetic risk information produces sustained changes in risk behaviour Predictive genomic medicine adopts a “high risk” strategy Population-based tobacco control strategies, such as taxing cigarettes and reducing opportunities to smoke, have halved cigarette smoking rates in Australia There are similar arguments for the greater efficiency of population-based strategies in reducing risky alcohol use, obesity, and diabetes. These strategies aim to reduce population access to cheap energy-dense foods and increase opportunities to exercise. Based on the successful experiences in tobacco control, such strategies will probably include: increased taxes on, reductions in the promotion of, and decreased availability of, energy-dense foods; and redesigning urban environments to reduce sedentary behaviour and increase opportunities for incidental exercise in everyday life Public health professionals are also concerned about the potential misuse of genetic risk information by industries that wish to promote harmful forms of consumption . These industries are likely to advocate for population-wide genetic screening in order to undermine public health policies that will reduce the use of their products in the population Analyses of industry documents The alcohol industry has also promoted the idea that alcohol-related problems only occur in a minority of genetically vulnerable drinkers The major public health challenge in formulating a policy toward population-wide genomic screening will be in discovering how to obtain whatever public health benefits genomic medicine delivers for common diseases without undermining effective population health policies that reduce exposure to the common risk factors responsible for the high prevalence of these diseases in developed countries Public health utility should, however, be differentiated from the usefulness of genetic information in a medical context. Research may identify low-frequency genetic variants with large effects that can be used in matching treatments to patients in clinical settings. Rare variants may also identify promising new targets for drugs to treat common diseases. But the usefulness of such variants for public health screening will be limited because of their low frequency in the population.Although early research has not shown a strong improvement in predictive power when genetic and environmental factors are combined to estimate disease risk, the use of Mendelian randomisation may enable epidemiologists to assess the causal role of environmental factors in common diseases But it is clear that genetic screening of whole populations is unlikely to transform preventive health in the ways predicted 10 years ago. The integration of individual genomic risk prediction into public health disease prevention strategies will require good evidence that this approach improves on the cost-effectiveness of existing population level interventions. The utility and cost-effectiveness of predictive genomics, like any other new health technology, should be evaluated disease-by-disease and population-by-population. Its utility will depend not only upon the costs of genetic screening  but also on: the effectiveness of treating those at increased risk; the morbidity and mortality that these preventive interventions avert and cause; and on our ability to prevent the subversive use of genomic information by interested industries to undermine effective public health policies. Until we have a much stronger evidence base, and more data on interactions between genotypes and common environmental exposures, advocates of genomic medicine should be much more modest than some have been in the claims they make about its likely impacts upon population health.


Ectomyelois ceratoniae  , is the most important pest of pomegranate in Iran. As it has been rarely recorded on other host plants, control methods have mostly been focused on its populations on pomegranate. In this study, shapes and sizes of wings were compared in populations on 4 host plants  using a landmark-based geometric morphometric method, and analysis of partial warp scores and centroid sizes. The results showed significantly smaller wing size in populations on pomegranate and a significant host plant-associated shape difference among populations as a consequence of allometric growth. This suggests that the wing size and shape differences among test populations may not have a genetic basis and could happen because of differences in the nutritional content of host plants. The results of the analysis suggest that the female carob moth lays her eggs on host plants that provide suitable conditions for hatching. The larger size of moths on hosts other than pomegranate showed that some host plants such as fig, pistachio and walnut can provide for increased stored nutritional reserves by larvae that may result in more successful over-wintering and higher fecundity in adults. This suggests that in spite of the more extensive activity of carob moth on pomegranate in Iran, populations on other host plants can have an important effect on expanding pest population sizes in following years which should be considered in control methods.The carob moth,  It is believed that nominally polyphagous species of herbivorous insects sometimes are comprised of multiple morphologically similar biological species with more specialized appetites . If hostEctomyelois ceratoniae  , the larva of which feed inside the fruit and highly affects fruit quality. This moth is commonly found in pomegranate orchards in Iran, but there are only a few records of its damage on other host plants such as fig  and Chatrood on pomegranate . From the province of Markazi: from the city of Saveh on fig  and pomegranate . From the province of Tehran: from the city of Eshtehard on walnut  , Table 1During the collections, infected fruits were collected and the larvae inside them were reared to adulthood in the laboratory. Wing slides were prepared from adults and examined using a dissecting microscope and a CCD video camera. To detect the genetic or environmental basis of host selection, some of the adults that emerged from infected pomegranates from Saveh, were transferred to pistachio and walnut to produce new generations that could be examined for differences between them. 17 forewings and 14 hindwings were obtained from individual moths reared on walnut  and pistachio .W matrix) as suggested by 10 landmarks on the forewing and 7 landmarks on the hindwing were chosen , and theA two-way MANOVA was performed to find significant differences among test populations and sexes, and to test whether the differences are the same in each sex. Morphologic distances among test populations were computed and the resultant distance matrixes were also subjected to cluster analysis by the unweighted pair group method to show similarity among test populations. In order to minimize the geographic effects (distances and barriers), and compare host plant associated populations, sympatric or quasi-sympatric populations on different host plants were compared: fig from Markazi province  versus pomegranate from the same site (MA-Po), pistachio from Kerman province  versus pomegranate from the same province , Walnut from Tehran province  versus pomegranate from Markazi province . Critical α were calculated using the Bonferroni method , hence tTo find any isometry in size variation between populations, analyses of allometry among known groups were performed. If rejected, there would be another question to be answered: Do the populations vary in different allometric trajectories? In other words, “Would shape vary significantly between populations if size were held constant?” Regression of shape on size was performed to find any allometry among pair-wise compared populations and then MANCOVAs were designed in any pair of compared populations using the method of fitting constants  to compaTo determine whether the size of wings in known groups varies significantly, ANOVAs on matrix of centroid size were performed. Statistical analyses were performed using NTSYS-pc  and MINITwo-way MANOVA showed the existence of significant differences among wing shapes in host plant associated and sexual populations, however interactions between them were not significant . The cluRegression of shape on size in the above comparisons showed significant allometric growth between sympatric or quasi-sympatric host plant-associated populations . AccordiComparing centroid sizes of host plant associated populations showed significant differences between them , and in all comparisons pomegranate associated populations had smaller wings than other host plant populations . CompariRhagoletis pomonella  are more similar to those from Chatrood , which are about 770 Km apart, than those on fig from the same site and also on walnut from Eshtehard which is much closer . Since nomonella  and 1993etua sp. . Alternanditions . This phE. ceratoniae was confirmed by further molecular analysis carried out by the authors (unpublished data). The molecular data obtained using 6 amplified fragment length polymorphism (AFLP) primer combinations showed high levels of variation within populations of E. ceratoniae, but non-significant genetic distances among sympatric host associated populations.Comparison of pair-wise geographically close populations on different host plants revealed significant shape variation in nearly all comparisons. Analyses of wing size showed that populations feeding on pomegranate have smaller wings than those feeding on the other host plants, even smaller than their own offspring that were reared on other host plants in laboratory. Allometric analyses showed allometric growth in test populations, i.e.; variation in size of specimen was accompanied with variation in shape and sympatric or geographically close populations on different host plants varied in the same allometric trajectory. Hence, significant shape variations between compared populations occurred because of the allometric consequence of size change, rather than having a genetic basis. A lack of genetic basis of variations between host associated populations of E. ceratoniae which emerges from early May in Iran, preferably attacks pomegranate first. Apparently this fruit provides suitable conditions for oviposition. This may be because of the physical condition of pomegranate neck that protects the eggs and leads larvae to the inside of the fruit. After completing some generations on pomegranate, as soon as other host plants provide suitable conditions for laying eggs, such as the grooves and tracks that occur on pistachio (istachio , some inistachio  showed tHeliconius erato, and it was shown that large flies have some advantages compared to small ones such as having higher mating fertilization and reproductive success (E. ceratoniae than infected pomegranates of the previous year. Hence, infected pomegranates may not be the only sites for over-wintering of E. ceratoniaes and collecting them may not effectively prevent over-wintering. Our observations while collecting specimens showed that high levels infestation by E. ceratoniae on host plants other than pomegranate were not seen. However, considering that our results suggest that infestation of other host plants is likely, it is necessary that control measures should not focus only on pomegranate, as alternate host plants could be important sites for increasing E. ceratoniae population in the future.It has been shown that the adult body size of some insects such as cerambycid beetles , dung be success  even if  success . In our


Lack of time and competing demands limit the ability of patients and providers to engage in informed decision-making discussions about prostate cancer screening during primary care visits. We evaluated a patient recall invervention to mitigate these challenges.Using mail and telephone outreach we invited men age 50-74 years without a PSA test in the prior 12 months to make appointments with their primary care providers in order to discuss the pros and cons of PSA-based prostate cancer screening. We assessed patient responsiveness to the program, provider documentation of screening discussions, orders for PSA laboratories, and provider attitudes.Out of 80 eligible patients, 37 (46%) scheduled and 28 (35%) completed a recall appointment. A large majority (91%) of patients eligible for PSA screening received an order for this test. Providers documented PSA discussions more often for these patients than for a recent sample of their other patients who received traditional care . Twelve of 14 participating providers felt the program improved their ability to impart information about the risks and benefits of screening, but were uncertain whether it influenced their patients' preexisting preferences for screening. Some expressed doubts about the advisability of PSA-specific appointments.To a limited extent, this pilot recall intervention enhanced opportunities for discussions of prostate cancer screening between patients and their primary care providers. As currently configured, however, this program was not found to be feasible for this purpose. A future version should promote screening discussions in the context of a broader range of health maintenance concerns and include more detailed, low-literacy information to educate patients in advance of clinic visits. Prostate cancer is the second-leading cause of cancer death in men over the age of 50. The prostate specific antigen (PSA) test combined with a digital rectal exam (DRE) can detect prostate cancer early; however, there is little high-quality evidence that screening and current treatments, many of which have disabling side effects, reduce mortality . For thiIn response to these challenges, efforts have focused on educating and activating patients for discussions through the use of decision aids and other educational materials. A few studies have demonstrated that decision aids are associated with increased knowledge and reduced screening rates ,8. PersoHere, we evaluate a modified approach to informed decision-making for prostate cancer screening. The approach involves using a registry to identify patients eligible for PSA screening and proactively outreaching to these patients by mail and telephone in order to schedule appointments specifically intended to facilitate meaningful discussions with medical providers about the risks and benefits of screening. The goal is to give patients the opportunity to consider a brief educational message at leisure in their home environment while providing ample time in clinic to discuss PSA screening with medical providers. We assessed the operational feasibility of the program; level of patient willingness to schedule and complete decision-making appointments; documentation of PSA screening discussions during clinic visits; rates of PSA ordering; and provider feedback about the program.This study was conducted in a large ambulatory primary care practice affiliated with the University of Colorado Hospital (UCH). The practice provides primary care to a diverse patient population  and is staffed by 40 attending physicians, four nurse practitioners, and 20 primary care residents. Attending providers were consulted and agreed ahead of time to participate in the pilot program described below.not have an administrative claim for a PSA test within the past 12 months (to coincide with standard recommendations for screening on a yearly basis if screening is to be carried out). Through a subsequent manual review process, men from this initial sample were excluded from outreach if they had clinic notes indicating an active cancer or a terminal diagnosis; were deceased; or no longer appeared to be receiving care within the system. In addition, men were excluded if PSA testing would not be warranted for screening purposes because of a history of prostate cancer or a previously elevated PSA result or prostate nodule. In other words, it is often not possible to determine eligibility based on automated (computer-based) EHR-review alone; a manual review of the EHR as well as patient return postcards and telephone contact are often necessary to identify patients who have received PSA screening outside the system in the specified time frame (12 months), who are no longer receiving care in the system (more likely if they inform us of same or have not received care in the system in >18 months), and who are ineligible for PSA screening. Such patients, once identified, are deemed ineligible and not offered phone-based appointment scheduling.Based on automated electronic health record  abstraction, men were deemed to meet initial inclusion criteria if they were age 50 to 74 years, had an attending (rather than resident) primary care provider (PCP), had seen a PCP in the practice at least once in the preceding 18 months (to minimize the inclusion of patients no longer receiving care in the system), and did Registry records of men meeting initial inclusion criteria were imported into an information management utility developed for this and other preventive and chronic disease outreach interventions -12. The whether to get prostate cancer screening." The letter encouraged the patient to contact our call center to schedule a PCP appointment to discuss PSA screening. A postage-paid return postcard accompanied each letter on which the patient could indicate whether he had recently had a PSA test outside of the health system, no longer received primary care within the system, was uninterested in discussing PSA screening, or preferred to be contacted at a specified time and telephone number. If the patient did not respond to the letter within two weeks by postcard or phone, an outreach coordinator (OC) made up to three calls to his home at various times of day, leaving a voice message on the first and last attempts. If a patient could not be reached after three calls had been made, he was considered a passive decliner of a PSA screening discussion.The information management utility generated invitation letters to be mailed to each patient's home. Each letter, bearing the name of the patient's PCP on the signature line, included the following message: "The PSA test can help doctors find and treat prostate cancer at an early stage, but the test is not perfect and does have potential downsides. Believe it or not, for example, there is still a lack of evidence that prostate cancer screening saves lives. Also, this type of screening can sometimes lead to unnecessary treatments that cause permanent side effects. Cancer prevention experts recommend that men speak with their doctors about the PSA test before going ahead with screening. The goal of a visit with your primary care provider is not to get prostate cancer screening but to help you make an informed decision about If telephone contact was established, the OC reviewed the rationale for having a discussion about PSA screening (i.e. men should know the pros and cons of prostate cancer screening before making a decision and the PCP will be able explain these). If the patient accepted, the OC scheduled a 20-minute appointment over the telephone. The OC then sent a brief note to the PCP via the EHR summarizing the purpose of the upcoming visit: "Patient X has scheduled an appointment with you at date/time to discuss prostate cancer screening. As you know, there are benefits and risks to such screening and it is important for men to be informed before making this decision. An informed decision-making reference is available at the following website: <url>" . FinallyWe assessed the following outcomes:These were characterized as the proportions of eligible participants who, during and after mail and telephone outreach, scheduled and completed an appointment with a PCP to discuss prostate cancer screening. Only those patients who scheduled an appointment with an OC through the call center , and who received verbal information from an OC about the informed decision-making purpose of the clinic visit, were considered to be responsive to the program.PSA discussion - generic reference to or specific details evidencing at least some screening "discussion" or "conversation"; (2) At least one downside - generic reference to having discussed, or specific documentation of, at least one downside of PSA screening; and (3) Patient preference - documentation that the patient did or did not wish to proceed with screening. For each provider who had at least one patient that completed a recall visit for the purpose of a screening discussion, we carried out a similar review of EHR documentation for the three most recent patients for whom the same provider completed PSA screening during a period immediately preceding implementation of the intervention. Based on USPSTF guidelines, these patients should  have engaged in a decision-making discussion before completing screening, which should have been documented in the EHR. We regarded these patients as having received traditional care. In other words, theoretically for this group, decision-making discussions could have taken place opportunistically during traditionally-arranged, episodic clinic visits . We then compared traditional and intervention care in terms of the presence of each type of documentation element.We determined the proportion of patients whose providers documented PSA screening discussions in the EHR during appointments scheduled for this purpose. Two research assistants also independently characterized the content of the clinic notes according to the presence or absence of three types of elements: (1) The rate of PSA ordering was characterized as the proportion of screen-eligible patients whose providers ordered or indicated an intention to order a PSA screening test during the clinic visit.We devised a brief online survey which we administered after the intervention to participating providers in order to assess their open-ended opinions about their willingness to discuss PSA screening with patients; the value they place on such discussions; the mechanics of the intervention; and perceived effects on quality of care.Statistical procedures were performed using STATA . Chi-square tests were used to determine the strength of association between patient characteristics  and willingness to schedule an appointment. Non-parametric Mann-Whitney tests were used to derive aggregate comparison measures of participating providers in terms of their average rates of including EHR documentation of screening discussion elements during traditional care and intervention-related visits. Descriptive statistics and quotes illustrative of recurrent themes were used to characterize the results of the provider survey.This intervention was designed and carried out as a quality improvement research program that relied on standard methods for creating patient registries and providing patient outreach. The Colorado Multiple Institution Review Board approved publication of results following the removal of personal health information.because of information contained in the outreach materials.Of the 154 patients who met initial, automated inclusion criteria, 40 (26%) were identified as meeting one or more exclusion criteria after brief manual review, leaving 114 in outreach sample. Characteristics of 80 patients deemed eligible for program participation after outreach was completed are summarized in Table patient's final screening preference  carry out prostate cancer screening discussions on their behalf, but one detractor suggested that this would only "increase the level of fragmentation" in patient care.Thirty-seven attending PCPs  who responded to the survey, including 14 that participated in intervention-related visits, agreed that discussions of the risks and benefits of screening are somewhat to very important. At the same time, however, two-thirds of respondents reported that such discussions are very challenging and rarely take place, even when extra time is made available. Said one provider, "Competing demands, time, and complexity of working through the hypotheticals are probably the biggest challenges. The near universal message in the media that men should have prostate screening also presents a challenge." Twelve of 14 PCPs who participated in intervention-related visits felt that their ability to share information about screening was enhanced, but most remained uncertain whether the extra time and information did much to sway their patients' Given the uncertain benefits and risks of harm associated with PSA-based screening, patients should participate in an informed decision-making process about this test . Howeverconversation about screening. For example, patients whose primary motivation was to complete screening might have been more likely to schedule appointments in order to complete screening. For these patients, discussions may have done little to change their minds . Meanwhile, patients with little interest in screening at the time of outreach, or after receiving a letter, might not have found the offer of a discussion particularly compelling and therefore may have declined to schedule one. On the other hand, we noted that half of these patients declined to schedule a visit because they had already made a decision to forego screening based on a prior conversation with their PCP about the topic. This supports the idea that such discussions can have an important influence on patient attitudes and behavior. A key question, then, is whether a patient recall program can motivate appointment scheduling among patients with ill-defined preferences for PSA screening.In a larger sense, however, it remains unclear whether the pilot program achieved its key objective, which is to help patients make genuinely informed decisions about screening. First, whereas we would have hoped that documentation of a discussion would have been evident in all of the intervention clinic visit notes, such documentation was present in less than half of these. Second, it is doubtful that all patients responded with equal enthusiasm to the offer of a These considerations, in addition to provider feedback about access problems in the clinic, suggest that a future recall program would be more effective if it facilitated PSA discussions within the context of addressing a larger set of health maintenance concerns. We did not wish to overwhelm patients with written information about prostate cancer screening, but earlier work suggests it may nonetheless be beneficial to include with the outreach materials more robust, low-health literacy educational materials or a PSA decision aid as these could help patients focus and be more prepared for discussions when they finally take place .We remain intrigued by a patient-centered medical home model of team-based care in which - instead of physicians - mid-level providers, including nurse practitioners and physician's assistants, assume greater responsibility for the provision of primary prevention services . With prnot order a PSA test. Nonetheless, we believe that where PSA tests were ordered, the intervention and traditional care differences in documentation are significant and likely to reflect meaningful differences in the frequency and quality of the decision-making process. Future versions of this intervention should improve accuracy in the eligibility review process (e.g. better identification and exclusion of patients with previously-elevated PSA results) as well as accord greater attention to variables that are likely to influence cost-effectiveness and long-term sustainability .While this study offers lessons for future work in prostate cancer screening informed-decision-making, it does have important limitations. The intervention included a relatively small number of patients, precluding meaningful subgroup analysis, and was performed in a single academic healthcare setting that is not representative of other types of practice environments. We did not assess patient knowledge, screening preferences, decision-making locus, and satisfaction before or after clinic visits. The lack of documentation of PSA discussions among some patients in the intervention group does not mean that such discussions did not take place. Similarly, it was not practically feasible to identify patients receiving traditional care who discussed PSA screening and whose providers, as a consequence, did intentions to be screened rather than provider orders for PSA tests. Finally, a recent systematic review indicates that only a very small number of interventions to educate and activate patients have been administered outside of clinical settings [Despite these limitations, this work had some strengths. First, under a low-risk clinical quality improvement umbrella, we were able to avoid selection bias by extending outreach to all men who were eligible for PSA screening rather than limiting participation to patients who formally consented to participation in a study. In contrast, prior studies of interventions to improve informed decision-making have limited participation to patients who provide written informed consent ahead of time ,18, who settings .In summary, this patient recall intervention appeared to activate and enhance opportunities for discussions of prostate cancer screening among male patients and their primary care providers, although not as frequently as desired. As currently configured, we cannot recommend this recall program for prostate cancer or other controversial screening tests. This type of intervention may be of greater interest and acceptability to patients if a future version promotes screening discussions in the context of a broader range of health maintenance activities and includes more detailed, low-literacy information designed to educate patients in advance of clinic visits.DRE: digital rectal examination; EHR: electronic health record; OC: outreach coordinator; PCP: primary care provider; PSA: prostate specific antigen; USPSTF: United States Preventive Services Task Force.The authors declare that they have no competing interests.TDD conceived the study, supervised intervention implementation and evaluation, and assumed primary responsibility for manuscript preparation. CTL and MB assisted in planning the intervention, reviewing the results, and preparing the final manuscript. AS, TM, and LS carried out the intervention and data collection. TM also completed the statistical analysis. All authors read and approved the final manuscript.Supported in part by funding by The American College of Physicians Foundation Health Literacy Awards Program.  and the American Cancer Society, MRSG-06-081-CPPB .The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2296/10/75/prepub


FTO) gene region with body mass index (BMI) and risk of obesity was identified in multiple populations, making FTO the first locus unequivocally associated with adiposity. At the time, FTO was a gene of unknown function and it was not known whether these SNPs exerted their effect on adiposity by affecting FTO or neighboring genes. Therefore, this breakthrough association inspired a wealth of in silico, in vitro, and in vivo analyses in model organisms and humans to improve knowledge of FTO function. These studies suggested that FTO plays a role in controlling feeding behavior and energy expenditure. Here, we review the approaches taken that provide a blueprint for the study of other obesity-associated genes in the hope that this strategy will result in increased understanding of the biological mechanisms underlying body weight regulation.In 2007, an association of single nucleotide polymorphisms (SNPs) in the fat mass and obesity-associated ( Obesity is therefore a major international public health threat and economic burden. Although environmental factors, such as little physical activity and over-eating, have driven the recent rise in the numbers of people who are overweight or obese, genetic factors are estimated to account for 40–90% of the population variation in BMI FTO) became the first to be associated reproducibly with human body mass. We review the implications of genetic association between SNPs in the FTO gene region and BMI in humans, the various studies undertaken and the challenges in progressing from a genetic association to new biological insight. We also provide an overview of possible future directions for research in this field.Overweight and obesity, defined as body mass indices (BMI) >25 and >30, respectively, are associated with premature death through increased risk of many chronic diseases, including type 2 diabetes, cardiovascular disease and cancer FTO region and risk of T2D FTO SNPs and T2D was mediated by an association with BMI FTO SNPs and BMI and the risk of being overweight or obese has been confirmed in multiple populations FTO SNPs on BMI is modest, with those individuals homozygous for the risk allele weighing, on average, 3 kg more than those homozygous for the protective allele Before 2007, despite huge efforts using genome-wide linkage studies and candidate gene association studies, no genetic variation had been unequivocally associated with BMI and risk of obesity in population studies FTO. Thus, the association signal could be due to correlation between FTO intronic SNPs and variation elsewhere in the gene or control elements of other genes. Indeed, the transcription start site of the retinitis pigmentosa GTPase regulator-interacting protein 1-like (RPGRIP1L) gene  is in close proximity (∼400 bp) to the 5’ end of FTOFTO are associated with obesity through an effect on RPGRIP1L. Efforts to fine-map the association signal to causative variant(s) are underway (Association does not necessarily mean causation. BMI-associated SNPs lie within a 47 kilobase (kb) linkage disequilibrium (LD) block encompassing parts of the first two introns as well as exon 2 of underway .FTO signal, additional GWAS have succeeded in identifying many additional novel obesity loci Since the discovery of the FTO and BMI were first discovered FTO gene product. Fto was first identified in the mouse by positional cloning Fto was only one of six genes deleted in the Ft mouse , it was not known which, if any, of these phenotypic manifestations were due to Fto deficiency. Interestingly, a phenotype similar to that of Ft mice was seen in a human patient harboring a small chromosomal duplication on 16q12.2, a region that includes the FTO gene When associations between SNPs at FTO. Moreover, FTO was not annotated in the public databases as having homology to any other known gene. The discovery that this mysterious gene was associated with human obesity In silico analyses of the human FTO sequence revealed homologs in other vertebrates  and marine algae (from unicellular photosynthetic picoplankton to multicellular seaweed) FTO shares features with Fe (II) and 2-oxoglutarate (2OG) oxygenases Escherichia coli enzyme AlkB and its eukaryotic homologs, which can repair DNA methylation damage by hydroxylating methyl groups on the DNA leading to their removal in vitro studies, which showed that murine Fto is indeed a 2OG oxygenase that can catalyse nucleic acid demethylation Clues from the Ft mouse and human chromosomal duplication were not sufficient to shed light on the specific effects of in vitro studies, as the activity of molecules might differ in whole organisms and under different conditions: therefore, animal models are used to further elucidate in vivo function.Caution is always recommended when interpreting gene function from Fto and human FTO mRNA expression showed that both are ubiquitous, with particularly high levels of expression in the brain and hypothalamus Fto mRNA expression in wild-type rodent tissues confirmed its ubiquitous expression, with high levels of expression in hypothalamic regions known to play important roles in the regulation of energy intake and expenditure, and suggested that expression of Fto might be regulated by nutritional status Fto mRNA expression compared to fed controls. This effect is not rescued by supplementation with the anti-starvation hormone leptin Fto expression observed during fasting is independent of leptin levels. In support of this conclusion, hypothalamic Fto expression is reduced in fasted Lepob mice (which lack leptin) compared to fed Lepob and control mice Fto is downregulated during fasting and upregulated during feeding, and that variation in Fto resulting in decreased expression or activity might provide a signal that promotes feeding and obesity.Early studies of mouse Fto expression was increased significantly in the hypothalamus of food-deprived and food-restricted rats Fto was widely and consistently expressed in brain regions related to circadian rhythmicity Fto in fasted rats suggests that high levels of Fto protein might stimulate food intake, which is the opposite effect predicted from its expression in wild type mice Fto mRNA in the hypothalamus decreased food intake, whereas a 40% decrease in Fto protein led to increased food consumption Fto were limited to a specific region of the hypothalamus (the arcuate nucleus), where the expression of Fto is very high. To date, all studies of rodents have been done over prolonged periods of fasting, which might better resemble starvation rather than fasting and hence some of the observations might reflect that. There is a need for studies with shorter fasting times in rodents to better understand the regulation of Fto expression during this time.In contrast to the mouse data, FTO are associated with human obesity and that Fto mRNA levels appear to be regulated in response to feeding and fasting, its expression was determined in relation to changes in obesity. In six mouse models of obesity ; Glossary), hypothalamic Fto expression did not differ significantly from that in wild type mice Fto gene expression in other metabolically relevant tissues, such as fat (adipose) tissue. Adipose tissue acts as a fat store and synthesizes and secretes a variety of proteins (such as leptin) that influence appetite and metabolism at distal sites. Expression of Fto in mesenteric fat was reduced significantly compared to wild type in all mouse models except DIO. This is interesting because murine Fto gene expression was shown to be downregulated under fasting conditions, suggesting that obese mouse models mimic the fasted state, possibly contributing to their over-eating.Given that variants in Fto expression differs under feeding and fasting conditions and displays tissue-specific differences in mouse models of obesity, but it is not known whether these differences are the cause or the consequence of obesity. To further investigate whether differences in Fto expression or function can cause increases or decreases in fat mass, mice harboring Fto mutations were generated.Fto deficiency have been reported: a null mutation (Fto−/−) resulting in the complete absence of Fto protein expression Fto mouse model carries a point mutation resulting in a change in the amino acid sequence from isoleucine to phenylalanine at position 367 (I367F). Although this residue is outside the catalytic core, it is located within a ∼20 amino acid block conserved throughout vertebrates that defines a new functional domain In vitro experiments demonstrated that, although the full-length I367F protein was correctly localized to the nucleus, it displayed lower levels of expression in mammalian cells and resulted in reduced catalytic activity, possibly through altering the Fto dimerization state. However, the exact role of different Fto dimerization states in energy balance is poorly understood. The mutant Fto I367F protein retains partial function, which is likely to account for some of the observed differences between the phenotypes of these two animal models. Namely, although both models demonstrated reduced body weight and fat mass, this starts early in life in the case of Fto−/− mice, whereas weight reduction in FtoI367F mice has a maturity-onset. Also, there is no discernible phenotype in the Fto null heterozygous mice, whereas the phenotype of the heterozygous FtoI367F is very similar to that of their homozygous mutant littermates. The importance of I367 in dimerization might help explain why the FtoI367F heterozygous mice have a phenotype: the I367F substitution might have a dominant negative effect by disrupting the function of wild type Fto through formation of heterodimers. The weight reduction observed in the two models is also very different: Fto null mice have a 30–40% weight reduction compared to wild type littermates, whereas the FtoI367F mutants have only a 10% reduction in weight. Another significant difference between the two models is that only Fto−/− mice exhibited growth retardation and early perinatal death Two mouse models of Fto mouse models is due to decreased energy intake, increased energy expenditure or both. Compared to wild type littermate controls, there was no difference in absolute food intake in Fto-deficient mouse models Fto−/− mice ate relatively more given their reduced body weight and size Fto-deficient mice was not due to reduced food intake. However, both models exhibited higher levels of energy expenditure (higher metabolic rate) such that overall the animals gained less weight and were protected from obesity FtoI367F mutant) but was potentially mediated by increased sympathetic nervous system (SNS) activity, a system that originates in the spinal cord with projections to peripheral tissues and is used by the hypothalamus to regulate energy homeostasis These studies then investigated whether reduced growth and adiposity in the FtoI367F mutants compared to wild type mice FtoI367F mutants demonstrated increased carbohydrate metabolism relative to fat metabolism. Hypothalamic neuropeptide expression did not differ between wild type and mutant in the fasted state, but Npy expression was lower in fed mutant mice. Given that increased Npy stimulates food intake, this finding suggests that Fto-deficient mice are more sensitive to satiety and are thereby protected from obesity through over-eating. A blunting of Npy induction was seen in Fto−/− mice in the fasted state, lending support to the suggestion that Fto promotes Npy expression Microarray analysis of white adipose tissue, liver and skeletal muscle detected some differences in gene expression in Fto−/− and FtoI367F mice support the idea that association of FTO SNPs with human obesity arises via regulatory or functional effects on FTO rather than other genes in the region. Although the Fto null mutation could impact regulation of other genes, the non-synonymous mutation in FtoI367F mice is most likely to exert its functional effects on Fto alone FTO and that inhibition of FTO might protect against obesity The phenotypic characterization of both the FTO was reported to segregate with an autosomal recessive disease in a large Palestinian Arab consanguineous family with nine affected members Fto mouse models. This discrepancy between the effects of human and mouse mutations  are eagerly anticipated. The lack of clinical obesity in patients carrying R316Q and family members is perhaps consistent with the relationship between loss of Fto function and leanness in mice. More recently, however, loss-of-function heterozygous mutations in FTO were found in both lean and obese humans, suggesting that haploinsufficiency does not protect against obesity completely, though the relationship might be complicated by other obesity-promoting factors Fto mutations and human FTO SNPs  could shed light on the biological mechanism impacting body mass, for example by determining whether the causal variant(s) change the amino acid sequence of FTO or whether they lie within a control element affecting expression of other genes in the region, such as RPGRIP1L. However, fine-mapping the association signal might be difficult because the obesity-associated SNPs lie within a 47 kb LD block in which the effects of causal variant(s) could be indistinguishable from other, highly correlated proxies. Under these circumstances it might prove more important to understand the biological effect of the risk haplotype  on genes and pathways. FTO obesity-associated SNPs do not appear to influence FTO expression but could be investigated for their effect on expression of its neighboring gene RPGRIP1L. Nevertheless, there is now some evidence to suggest that the associations with BMI are indeed mediated through FTO . Though none of the obesity-associated SNPs fall within these elements, the possibility that the association signal results from correlation with causal variant(s) within these regions has not been excluded At present, the strongest associations between ough FTO ; howeverFTO has generated many questions as well as answers. Perhaps the most important lesson from the extensive work on FTO over the last three years is the promise that other obesity loci identified by GWAS could lead to new understanding of the underlying biology of human adiposity. Despite the small effect sizes and predictive value of the common variants identified to date, these hits often represent previously unsuspected obesity genes. The story of FTO research provides valuable lessons for the study of other obesity-associated genes. Ultimately, it is hoped that knowledge of susceptibility gene function will highlight novel genes and molecules that could be targeted for therapeutic intervention. For example, the data presented in this review indicate that FTO deficiency protects against obesity. However, the devastating effects of a loss-of-function non-synonymous mutation in FTO in a human family emphasizes that care should be taken in developing drugs to alleviate obesity by reducing the expression or inactivating catalytic activity of FTOAy: the lethal yellow (Ay) mutation in the agouti gene produces mice with a complex phenotype including obesity and a yellow coat color Body mass index (BMI): weight (in kilograms) divided by the square of height (in meters). Overweight is defined as BMI >25, and obesity as BMI >30.Cepfat: homozygous loss-of-function mutations in the carboxypeptidase E gene result in obesity and hyperglycemia Diet-induced obesity (DIO): mice with obesity induced by diet, rather than purely genetic causes Dominant negative: a mutation that results in a protein that interferes with the function of the wild type protein, usually through dimerization with the wild type protein.Energy balance: energy intake versus energy expended as internally produced heat and physical activity.Lepob: disruption of both copies of the leptin gene results in obese, hyperphagic (over-eating) mice Leprdb: shares similar features with the Lepob mouse and results from abnormal splicing of the long, hypothalamic leptin receptor Linkage disequilibrium (LD) block: linkage disequilibrium is the name given to the phenomenon whereby alleles at multiple loci appear together more often than would be expected by chance. This is often measured by r2, which ranges from zero (unlinked loci) to 1 (complete correlation between loci). An LD block defines a region within which variable loci are highly correlated .Lipolysis: the breakdown of fat (triglycerides) into free fatty acids within body cells.Polydactyly: having >10 fingers or toes.Tagging SNPs: SNPs that can be used as proxy for other, correlated SNPs ).Thermogenesis: heat production. A component of energy expenditure that can be stimulated by the sympathetic nervous system.Tub: tubby mice exhibit adult-onset obesity due to mutation in the Tub gene It is clear that the journey undertaken by


Mitotic cell death is an important form of cell death, particularly in cancer. Chromosome fragmentation is a major form of mitotic cell death which is identifiable during common cytogenetic analysis by its unique phenotype of progressively degraded chromosomes. This morphology however, can appear similar to the morphology of premature chromosome condensation (PCC) and thus, PCC has been at times confused with chromosome fragmentation. In this analysis the phenomena of chromosome fragmentation and PCC are reviewed and their similarities and differences are discussed in order to facilitate differentiation of the similar morphologies. Furthermore, chromosome pulverization, which has been used almost synonymously with PCC, is re-examined. Interestingly, many past reports of chromosome pulverization are identified here as chromosome fragmentation and not PCC. These reports describe broad ranging mechanisms of pulverization induction and agree with recent evidence showing chromosome fragmentation is a cellular response to stress. Finally, biological aspects of chromosome fragmentation are discussed, including its application as one form of non-clonal chromosome aberration (NCCA), the driving force of cancer evolution. Morphological characterization of abnormal chromosomes represents an important aspect of medical genetics. Modern medical genetics was firmly established following the successful identification of trisomy 21 in Down syndrome ,4, but sDespite decades of intensive effort, identification of commonly shared chromosome aberrations for most diseases has been difficult -9. In coChromosome fragmentation is one type of recently identified NCCA that requires more attention . ChromosThe identification of chromosome fragmentation as a form of cell death is new to the field, but the morphological description of chromosome fragmentation is not. With similar morphologies, chromosome pulverization and PCC have been extensively studied since the 1960s ,15. ManyChromosome fragmentation is a major form of mitotic cell death that is identified through abnormal cytogenetic figures ,13. ThesChromosome fragmentation is non-apoptotic as evidenced by differences in morphology and other biochemical differences . ApoptosPCC was first described in 1970 by Johnson and Rao, using cellular fusion methods developed previously ,17. PCC PCC results in three distinct chromosomal morphologies determined by the cell cycle progression of the induced cell ,23. G2 PFusion and chemical inhibitor based methods of inducing PCC are similar in molecular mechanism and morphology. Both methods result in exposure of an interphase cell to the activated cyclin b/cdk1 complex  . In fusiPCC has applications in the research lab and the clinic. Chemical induction of PCC can be used to increase the number of mitotic cells within tumor samples, allowing for rapid karyotyping with only very short term culture being required . PCC canMultiple reports analyze two phenomena called chromosome pulverization or chromosome shattering -32. ThesPrevious reports have shown chromosome fragmentation but have called it chromosome pulverization/shattering, therefore one option is to continue to use the term(s) shattering/pulverization, however there are a number reasons not to do so. They include the following: First, descriptions of pulverization have focused largely on identification of later stage fragmentation events. The term chromosome fragmentation encompasses all stages of this mitotic cell death while pulverization has only been used to describe late stages. Second, DNA fragmentation is used to describe the genome degradation that takes place during apoptosis . TherefoChromosome fragmentation and PCC can be confusingly similar, so what exactly differentiates these phenomena? In the following discussion, chromosome fragmentation and PCC will be differentiated first by morphology and then by mechanism Table .S-phase PCC and chromosome fragmentation each produce striking mitotic figures where gaps/breaks are interspersed among pieces of condensed chromosomes ,14,34. SMost mitotic figures from cells undergoing chromosome fragmentation show variable degrees of fragmentation across the chromosomes where chromosomes that are completely degraded are found within figures that also contain completely intact chromosomes and every degree of degradation between these extremes also can be seen Figure . In humaMultinucleated and micronucleated cells are an interesting case where chromosome fragmentation and PCC can occur and affect a limited amount of the genome within the multinucleated cell. Multinucleated cells are a common occurrence in tumors, indicative of genomic instability, not commonly shared and are therefore one type of NCCA ,39. In mIt should be pointed out that chromosome fragmentation is a unique phenomenon of mitotic cell death, and should not be confused with just any type of chromosomal breakage. Chromosome fragmentation involves chromosome breaks, but this does not mean they are the same. For example, the recently reported phenomenon that there appears to be chromatin tethers that hold broken chromosomes together is likely not associated with chromosome fragmentation . It woulFurther work has also separated the two distinct phenomena. Chromosome fragmentation was not inducible in double thymidine blocked S-phase cells that remained in interphase (Stevens unpublished observation). This indicates that these cells arrested properly in either S-phase and/or G2 and were not able to prematurely enter mitosis. PCC, when the morphology is similar to chromosome fragmentation, occurs specifically in cells in S-phase . InterspPCC results in the abnormal activation of MPF, so inhibition of this activation would inhibit PCC. Roscovitine is a purine analog that functions as an inhibitor of the G2 to M transition through inhibition of CDK1 activity ,42. Roscin vitro studies and is now in active clinical trials [Though chromosome fragmentation and PCC can typically be differentiated based on morphological and mechanistic differences there is potential overlap of the two. This potential overlap occurs at the G2 checkpoint where any potential DNA damage is repaired prior to mitosis. Induction of PCC occurs through the abnormal activity of MPF subsequently subverting the G2 checkpoint. Traditionally, caffeine has been used to inhibit the G2 checkpoint and concurrent treatment with various drugs or radiation can result in chromosome fragmentation ,43-45. Ul trials -49. Concl trials . Chromosl trials . In thisMitotic catastrophe is often used to refer to mitotic related cell deaths. However there is no distinct definition of this type of cell death. Reports vary widely and include deaths that occur directly during mitosis, or occur following a failed mitosis -56. Due Certain models of mitotic catastrophe, particularly those that are based on cell fusion, are in fact suggestive of PCC. However others are based on loose G2 checkpoints ,51. Whenin vitro model of spontaneous transformation, p53 function is abrogated in the germ line [Following the discussion of the importance of chromosome fragmentation in cell death, it is necessary to mention its importance in genome research as well as future diagnostic use. Importantly chromosome fragmentation is one manifestation of the previously ignored non-clonal chromosome aberrations. NCCAs reflect the level of genomic instability within a population of cells, which according to the principles of the genome theory, are the driving force of cancer evolution and drug resistance -12,59,60Increased chromosome fragmentation has also been shown to occur in the PA-1 cell line where chromosome fragmentation takes place in order to eliminate polyploidy cells . ChromosAlthough chromosome fragmentation is important to maintain system stability, there are many redundant systems within cells that ensure that cells maintain homeostasis and are not permanently damaged at the genome level. Chromosome fragmentation becomes activated when cells reach mitosis with extensive damage or in the face of extensive stress. Chromosome fragmentation works in conjunction with other regulators of cellular integrity, such as the cell cycle checkpoints and other forms of programmed cell death (apoptosis and autophagy), to maintain the cellular system identity. Therefore, chromosome fragmentation can be applied as a clinically useful measure of NCCAs. Furthermore, chromosome fragmentation frequencies have also been shown to roughly approximate total NCCA levels. This is significant, as chromosome fragmentation analysis can be used as a relatively quick and inexpensive substitute for spectral karyotyping in patient monitoring. Although chromosome fragmentation indices may not hold all the information that SKY analysis could reveal, it does give an overall estimate of the genomic instability contained within a given sample. This information combined with information from the same slides such as mitotic index, frequency of apoptotic cells, and frequency of micronuclei may be useful in determining treatment modalities in many diseases including cancer.Chromosome fragmentation is a form of mitotic cell death that was only recently characterized. In some manifestations, chromosome fragmentation can appear quite similar to PCC. However in depth analysis shows that chromosome fragmentation and PCC are in fact quite different phenomena, though at times they may be linked to each other. They are distinguishable by morphological and mechanistic analyses. Distinguishing between chromosome fragmentation and PCCs is significant. Demonstration that they are two independent biological phenomena means that chromosome fragmentation can now be used as an easily applied cytogenetic index to measure mitotic cell death that could have clinical and basic research applications. In addition, despite decades of extensive studies, there is little known about the biological significance of chromosome pulverization. By separating chromosome pulverization from PCC, a long standing puzzle can now be solved. It now seems that the biological meaning of PCCs is rather limited. Well controlled experiments are needed to further separate PCC and chromosome fragmentation. In addition, chromosome fragmentation has been identified as a form of NCCA, which is the driving force of cancer progression, and therefore analyses of chromosome fragmentation in clinical settings could lead to better treatments and outcomes in cancer therapy.BrdUrd: Bromo-deoxyuridine; MPF: Mitosis promoting factor; NCCA: Non-clonal chromosome aberration; PCC: Premature chromosome condensation; SCE: Sister chromatid exchange; SKY: Spectral karyotypingThe authors declare that they have no competing interests.JBS and HHH wrote the paper. SMR contributed table


Dosage sensitivity is an important evolutionary force which impacts on gene dispensability and duplicability. The newly available data on human copy-number variation (CNV) allow an analysis of the most recent and ongoing evolution. Provided that heterozygous gene deletions and duplications actually change gene dosage, we expect to observe negative selection against CNVs encompassing dosage sensitive genes. In this study, we make use of several sources of population genetic data to identify selection on structural variations of dosage sensitive genes. We show that CNVs can directly affect expression levels of contained genes. We find that genes encoding members of protein complexes exhibit limited expression variation and overlap significantly with a manually derived set of dosage sensitive genes. We show that complexes and other dosage sensitive genes are underrepresented in CNV regions, with a particular bias against frequent variations and duplications. These results suggest that dosage sensitivity is a significant force of negative selection on regions of copy-number variation. The opposing theory could be called the selection hypothesis, stating that negative and positive selection shape the distribution of CNVs through the functional elements they encompass.It has been estimated that at least 2% of the human genome is affected by structural variations heterozygous mutations et al.dosage sensitive.Gene duplication and loss are key mechanisms in evolution et al.balance hypothesis). A range of experiments lend support to the balance hypothesis. It has been noted that expression levels of interacting proteins are highly co-ordinated What are the underlying causes of dosage sensitivity? Papp et al.et al.et al., and were not significantly enriched for protein complexes.There have been, however, several conflicting reports. Deutschbauer master-regulators controlling the balanced expression of a range of other proteins These findings point towards a more complex relationship between haploinsufficiency and duplication sensitivity The newly developed CORUM database via BioMART.The CORUM database As an alternative to the manually compiled set of complexes in CORUM, an independent set of putative complexes was computationally derived from high-throughput protein interaction experiments by identifying highly connected clusters of proteins in an extended network of human protein interactions Dosage sensitive genes were extracted from the annotations of the Baylor College of Medicine Medical Genetics Laboratory 105k diagnostic Chromosomal Microarray (version 7) A separate set of genes overexpressed in cancer tissue was also used et al.et al., there is a correlation between detectable expression variation and total expression strength  Segmental duplications of Gene Ontology (GO) annotations from the GOA project dN/dS values for human genes relative to mouse orthologs were acquired from EnsEmbl via BioMart. Only genes with a single unique ortholog in mouse were used in the analyses.In-species paralogs for 10755 HGNC gene identifiers were downloaded from EnsEmbl Compara via BioMart. The paralog prediction uses automatically generated maximum-likelihood phylogenetic trees. Details can be found at the EnsEMBL compara website.Statistical significance of overlaps between gene sets was computed with Fisher's exact test (FET). The Mann-Whitney-U test (MWU) was employed to determine significance of differences between two distributions. In cases of multiple testing, Bonferroni correction was applied. All calculations were performed in R cis and trans effects of copy-number variations (CNVs) on genes. However, there are few reports of a direct quantitative effect on expression levels for genes inside a specific CNV et al.Association studies We find that duplications and deletions have distinguishable profiles of expression ratios. The expression ratio is defined as the average expression of a gene in individuals with a CNV phenotype, divided by the average expression in wild-type individuals. Assuming a simple linear relationship between copy-number and expression level, one would expect a distribution with peaks at The magnitude of the expression difference between CNV and wild type individuals is smaller and more continuous than expected. However, the location shift between the two distributions is highly significant , defined as the average variation in absolute expression levels per gene, normalised to the mean absolute expression level, see et al.Axelsen CORUM is a manually curated data source and thus prone to ascertainment bias. To ensure that these results are not biased by the composition of CORUM, we generated a separate dataset of putative protein complexes extracted from several high-throughput protein interaction detection experiments see . The cluWe have shown above that protein complexes are under constraint to maintain their relative expression levels and exhibit limited expression variability between individuals. For our further analysis of dosage sensitivity, we also used an independently assembled set of 146 genes with known dosage-related disease phenotypes see . There iAs previously stated, we found that CNVs can affect the expression levels of genes they contain. We therefore hypothesised that a CNV that encompasses a gene which is part of a protein complex will be more likely to have a negative effect on fitness. As most available CNV data were derived from healthy individuals, we expect that genes encoding protein complexes will be underrepresented in CNV regions.Out of 18534 protein coding genes for which both genomic locations and a unique gene name could be retrieved, 2311 genes are fully inside a CNV region. From 1975 proteins in the CORUM database, only 165 are found in a CNV region, significantly fewer than one would expect by chance  category “binding” within CNV regions, but they do not comment further on this fact. We verified independently that “binding” is the second most underrepresented GO category after “intracellular” amongst genes in CNV regions. This lends further support to the hypothesis that dosage sensitivity due to protein complex membership has an influence of the composition of CNV regions.Nguyen We speculated that a negative fitness effect due to a copy-number variation will increase the likelihood of subsequent removal of that CNV from the gene pool. The CNVs that contain CORUM genes occur in significantly fewer individuals  are of fundamentally similar nature and frequently overlap. We thus hypothesised that the reduction in negative selection within CNVs is related to the higher copy number of some genes which have been recently duplicated in a fixed SD. If we split the genes in CNV regions into those that overlap a SD and those that do not, we see that dN/dS ratios are highly significantly elevated in the genes that overlap SDs  are biased towards deletions rather than duplications. If we assume that negative selection is acting on CNVs, the intuitive biological explanation for this phenomenon would be that CORUM genes are at least as sensitive to duplication than deletion, which in turn supports the concept that members of protein complexes are sensitive not just to under- but also to overexpression.We made another observation that support this hypothesis. When comparing a manually curated set of dosage sensitive genes derived from the scientific literature, we found that a significantly larger than expected proportion of these genes were members of a protein complex as defined by the CORUM database. Taken together, these findings clearly indicate that stoichiometric fluctuations negatively affect protein complexes.A key proposition that underpins our understanding of dosage sensitivity is that duplication or deletion of the genomic region containing a gene will result in a significant up- or downregulation of expression of the gene e.g. increasing or decreasing transcription or modulating mRNA degradation. Such autosomal dosage compensation has been first observed in maize and DrosophilaWe speculate, however, that there could also be a physiological explanation for the unexpectedly low change in gene expression upon copy-number variation. It is conceivable that the cell attempts to compensate changes in copy number on gene expression by It has been noted Interestingly, we also show that genes in CNV regions have significantly more paralogs than expected by chance, while genes in protein complexes possess, on average, fewer paralogs Conversely, the reason for the increase in dN/dS in many genes within CNV regions could be attributed to their higher number of paralogous sequences. In fact, genes in CNVs overlapping segmental duplications are again primarily, but not exclusively, responsible for the elevated dN/dS ratios. These observations underline that CNV regions are a frequent source of gene duplicates which occasionally get fixed over the course of evolution and thus drive evolution of some gene families.We observed that CNV regions are less likely to contain genes encoding protein complexes, as well as other dosage sensitive genes. Furthermore, CNVs which occur in multiple individuals and can thus be assumed to be older than unique CNVs are particularly depleted of CORUM genes. Hence, it appears that pressures on correct dosage limit the set of genes which can sustain variation in copy-number, even though the effect of CNVs on gene expression is not straightforward.et al.Dang


EHP 116:190–195; Parvez et al.][.Exposure to toxic levels of arsenic is a serious problem in many Asian countries, notably Bangladesh and India, where exposure to inorganic arsenic through naturally contaminated groundwater is widespread and often excessive. Scientists have long recognized the lung to be a major site of action of ingested arsenic, and most of the focus has been on risks associated with lung cancer. Nonetheless, habitual ingestion of high-arsenic drinking water also promotes many noncancer lung effects, and researchers are investigating the potential utility of biomarkers to identify such effects One of the challenges of pursuing connections between arsenic and respiratory illness is that assessments of respiratory symptoms may be prejudiced by interviewer bias if study participants show visible skin lesions, a hallmark sign of chronic arsenic poisoning. To overcome this obstacle, researchers have begun exploring the use of biomarkers for chronic respiratory disease such as serum levels of Clara cell protein CC16. This serum marker runs low in individuals with compromised lung conditions induced by chronic environmental exposures such as cigarette smoking or ozone.In the current study, investigators sought to determine the relationships of serum CC16 with well-water arsenic, total urinary arsenic, and urinary arsenic methylation indices in a population of 241 nonsmoking individuals exposed to arsenic-laden drinking water in Araihazar, Bangladesh. The mean arsenic concentration in drinking water was 134 μg/L, and individuals with skin lesions consumed significantly more arsenic than those without .In individuals with skin lesions (but not those without such lesions), there was a significant inverse association of CC16 with urinary arsenic as well as a marginally significant inverse association of CC16 with the cumulative arsenic exposure index, which estimates long-term exposure. The researchers speculate that individuals with skin lesions either are exposed to higher levels of arsenic or have a unique susceptibility to the respiratory effects of arsenic exposure for reasons unknown.The analysis also revealed positive associations of CC16 levels with the secondary arsenic methylation index (an indicator of arsenic methylation capability), particularly among individuals without skin lesions. This suggests that individuals with better methylation capacity may be less susceptible to the adverse respiratory effects of arsenic. Moreover, the inverse association of CC16 with the percentage of mono-methylated arsenic (a commonly measured arsenic metabolite) in urine indicates that individuals with incomplete methylation may be more vulnerable to arsenic-induced respiratory problems.This cross-sectional investigation, the first to use biomarkers of arsenic exposure and lung injury in this way, is an important step toward further improving our understanding of arsenic-related non-malignant respiratory illness. Serum CC16 shows promise as a biomarker for assessing early respiratory damage induced by arsenic, especially among individuals with skin lesions associated with the consumption of arsenic-contaminated drinking water.