The cerebellum regulates complex movements and is also implicated in cognitive tasks, and cerebellar dysfunction is consequently associated not only with movement disorders, but also with conditions like autism and dyslexia. How information is encoded by specific cerebellar firing patterns remains debated, however. A central question is how the cerebellar cortex transmits its integrated output to the cerebellar nuclei via GABAergic synapses from Purkinje neurons. Possible answers come from accumulating evidence that subsets of Purkinje cells synchronize their firing during behaviors that require the cerebellum. Consistent with models predicting that coherent activity of inhibitory networks has the capacity to dictate firing patterns of target neurons, recent experimental work supports the idea that inhibitory synchrony may regulate the response of cerebellar nuclear cells to Purkinje inputs, owing to the interplay between unusually fast inhibitory synaptic responses and high rates of intrinsic activity. Data from multiple laboratories lead to a working hypothesis that synchronous inhibitory input from Purkinje cells can set the timing and rate of action potentials produced by cerebellar nuclear cells, thereby relaying information out of the cerebellum. If so, then changing spatiotemporal patterns of Purkinje activity would allow different subsets of inhibitory neurons to control cerebellar output at different times. Here we explore the evidence for and against the idea that a synchrony code defines, at least in part, the input–output function between the cerebellar cortex and nuclei. We consider the literature on the existence of simple spike synchrony, convergence of Purkinje neurons onto nuclear neurons, and intrinsic properties of nuclear neurons that contribute to responses to inhibition. Finally, we discuss factors that may disrupt or modulate a synchrony code and describe the potential contributions of inhibitory synchrony to other motor circuits. Purkinje neurons are the principal neurons of the cerebellar cortex Figure . They reA primary question in cerebellar physiology, therefore, is how cerebellar nuclear cells transduce input from Purkinje cells to generate cerebellar output Figure . Becausetop). Consistent with this idea, nuclear cells often respond to sensory inputs with reduced spike rates . This idea is attractive, especially given the high densities of low-voltage activated “T-type” Ca currents in nuclear cells  recruit inhibitory inputs to that Purkinje cell, by activating molecular layer interneurons  receptor agonists, however, left synchrony unaffected, leading to the conclusion that Purkinje–Purkinje inhibition, mediated by local collaterals, provided the GABAergic signals that were crucial to maintaining synchrony. This idea is consistent with computational work demonstrating that synchrony is an extremely common consequence of synaptically connected oscillatory inhibitory cells  boutons all in contact with the same cell body.” Ramón y Cajal similarly observed that each Purkinje neuron axon formed six to eight “nests” onto as many cells  of behaving monkeys, spike synchronization is observed during voluntary movement, even without significant changes in firing rates, giving rise to the notion that dynamically organized cell assemblies are involved in generating movement  beta frequency (10–15 Hz) oscillations selectively predict motor performance in a coordinated arm and eye movement task in monkeys, leading to the suggestion that LFP oscillations help coordinate activity in distant, distinct cortical areas that control arm reaching and saccadic eye movements (Dean et al., Two major aspects of the hypothesis that the degree of Purkinje cell synchrony affects nuclear cell output, which we refer to as “synchrony coding,” are well supported by evidence: first, Purkinje cells indeed synchronize their simple spikes during behaviors, and second, the biophysical specializations of nuclear cells are well suited to permit entrainment to synchronized IPSPs. Nevertheless, to what extent, if at all, and under what conditions, if any, a time-locked response to synchronous IPSPs is a significant mechanism for encoding cerebellar output has yet to be demonstrated. At the level of the cerebellar cortex, specific remaining questions are how many Purkinje cells actually synchronize in response to specific stimuli, and how many of these cells converge onto common targets. At the level of the nuclei, major questions are whether the naturally occurring fractional synchrony is sufficient to engage time-locking, and how concomitant excitation or neuromodulatory input shapes the response to synchronized inhibition.None of these questions need have unique answers. The likelihood of simple spike synchrony, for example, may depend on the mode of firing by Purkinje cells. Analyzing data from awake and anesthetized rodents, De Schutter and Steuber  noted boRegarding regional variation, synchrony of Purkinje cell simple spikes indeed appears to differ across cerebellar cortical areas. In crus I and II, synchrony is more difficult to detect (Heck et al., A receptor kinetics on synchrony has been shown at other synapses. In the hippocampus, the kinetics of GABAA receptor mediated currents determine in part the rate at which the network can oscillate, such that rapid decay time constants support high frequency oscillations (Wang and Buzsáki, Regarding alternative mechanisms to synchrony coding, recordings from rat cerebellar slices have presented arguments for linear processing in the cerebellar cortex, by illustrating the linearity of input-output relations of several cerebellar synapses, including parallel fiber synapses and inhibitory synapses onto Purkinje cells (Walter and Khodakhah, A parallel example of spike timing shaped by inhibition is evident in the basal ganglia output to the thalamus, an anatomical and physiological sister circuit of the corticonuclear pathway. Spontaneously active, GABAergic internal pallidal neurons, like Purkinje neurons, often show overall increases in firing rate during target thalamic neuron activation (Anderson and Horak, Resolving whether and when synchrony coding is useful or necessary to cerebellar function will depend in part on understanding what nuclear cells communicate to down-stream targets. In the red nucleus, individual interpositus axons ramify parasagittally (Shinoda et al., The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Epithelial ovarian cancer (EOC) continues to represent one of the most lethal conditions in women in the western countries. With the shifting of childbearing towards higher age, EOC increasingly affects women with active childbearing wish, resulting in major impacts on treatment management. Next to the optimal therapeutic treatment strategies, gynecologic oncologists are being asked to incorporate into their decision-making processes the patients' wish for fertility preserving alternatives ideally without compromising oncologic safety. Nowadays, fertility-sparing surgery represents an effective alternative to conventional radical cytoreduction in younger women with early stages of the disease. As such, this paper considers indications for fertility sparing surgery in EOC, reflects on outcomes from the oncologic and reproductive data of the largest and most relevant series outcomes data, reporting on fertility sparing techniques in EOC, reviews medicamentous efforts to prevent chemotherapy induced gonadotoxicity, and discusses future aspects in the gynecologic cancer management. Epithelial ovarian cancer (EOC) continues to represent a lethal condition which commonly affects women in a multifocal and peritoneal metastasized fashion. Attributed to a special tumor biology and large heterogeneity, clinical outcomes in EOC vary broadly and although significantly associated with an adequate systemic and operative treatment, they often detach themselves from these common and known influential factors and follow not yet fully understood tumor biology patterns .With the constant shifting of childbearing age towards higher ages, the increasing incidence of EOC in women with active childbearing potential constitutes a therapeutic dilemma . Both paEven though radical surgery with the primary objective of maximal tumor reduction is currently the established cornerstone in the management of advanced EOC , fertiliNevertheless, the inevitable question arises, whether fertility-sparing surgery (FSS) for EOC harbors life-threatening risks, which compromise patient's survival and set any chance for a normal family life into the background due to early and potentially chemotherapy-resistant relapse.Since no randomized trials exist or will ever exist to prospectively evaluate and answer this question, our experience is limited to scattered, retrospective cases series. The aim of this paper is to provide an overview of the most significant hitherto reports and discuss future perspectives.FSS for ovarian neoplasms have been traditionally adopted in early-stage malignant ovarian germ cell tumors and in ovarian sex cord-stromal tumors such as granulosa-cell tumors, Sertoli-Leydig cell tumors, ovarian dysgerminomas, as well as in borderline tumors of the ovary with excellent reproductive outcomes without compromising oncologic safety –9. Gold For invasive EOC data are much more constricted and limited to retrospective, nonrandomised series referring mainly to patients with low-grade stage IA tumors with favorable histology, while data regarding higher stages of the disease or unfavorable constellation of histologic characteristics are rather conflicting , 10–12. According to the 2007 guidelines of the American College of Obstetrics and Gynecology (ACOG), fertility-sparing surgery for reproductive-age patients with invasive EOC is recommended for highly or moderately differentiated stage IA disease with non-clear-cell histology . In an eOpen questions remain if there should be differentiation between iatrogenic Ic disease due to intraoperative tumor rupture versus Ic due to tumor on ovarian surface or malignant cells in peritoneal cytology, whether patients with G3 tumors with no evidence of further metastatic disease in adequate staging are eligible of FSS and whether nonserous or non-endometrioid histologic subtypes should be a priori excluded from any organ-preserving technique.Data from the largest report series in the literature concerning oncologic outcome after FSS in EOC are summarized in When collectively evaluating most published results so far, mean relapse rates are estimated to be around 10%, even in patient's cohorts which included also Ic stage disease , 20–23. In the study of Zanetta et al., none of the women undergoing bilateral oophorectomy had microscopic foci of cancer in the normal-looking contralateral ovary. The rate of bilaterality is congruent with the previous observations of other authors who reported an extremely low rate of contralateral involvement for mucinous tumours .In two very recent studies originating also from the same authors group by Kajiyama et al., the authors assessed the feasibility of fertility-sparing surgery in patients with clear cell or mucinous carcinoma of the ovary, two histological types which have been associated in various reports with a rather less favorable prognosis , 22. In When attempting to identify a recurring profile of the patients who relapsed after FSS, then no specific pattern can be identified, since in various series also Ia patients tend to recur, whereby here one has to consider the possibility of occult more advanced disease after inadequate staging. One of the major limitations of the existing studies is that not all of the cases underwent an obligatory systematic lymphadenectomy, in which case there might occur an up-staging of some patients , 16. RepThe issue of impaired fertility in young cancer survivors represents a therapeutic dilemma for both treating physicians and affected patients. Systemic and operative oncologic treatments consistently compromise the ovarian function, often resulting in infertility and premature menopause –28.Data regarding reproductive outcome after FSS in EOC are summarized in When evaluating the incidence of spontaneous abortions, then rates range between 11% and 33%. No evidence is however given regarding details about the gestational week, the onset of symptoms, and whether there was a habitual recurring modus, so that no conclusions can be extracted about the cause of the abortions and their pathophysiology.In the recent evaluation by Satoh et al. , there aWherever reported, none of the patients who successfully conceived and gave birth presented any relevant, cancer-related clinical problems during the perinatal period. Also no higher rates of congenital malformations or abnormal fetal outcomes have been reported in the current literature , 16, 32.It is well known that the number of oocytes decreases as a normal process from the fetal life up to menopause. At 20 weeks of gestation, female infants have about six to seven million oocytes, newborns only one to two million, and women at the age of 37 have only about 25,000 oocytes left. Chemotherapeutic agents, which act cytotoxic by interrupting the normal cell cycle and inducing apoptosis do also negatively affect the highly endocrine-active ovarian tissue. Cisplatin and its analogues, that is, agents which play a highly significant role in the management of ovarian cancer, have been proven to present a risk factor for ovarian failure by an estimated odds ratio as high as 1.77 . In ordeThe possibility of reproductive dysfunction as a consequence of cancer treatment has an established negative impact on the quality of life of cancer survivors . The merAfter thorough insight of the current literature, FSS in early-stage EOC appears an absolutely viable and safe option for women younger than 40 years who wish to preserve their childbearing potential after careful consideration of histologic subtypes. The optimal indication is referring to stage Ia G1/G2 disease, as well as stage Ic with favorable, that is, non-clear-cell histology. Here there has to be differentiated between iatrogenic—due to intraoperative tumor rupture—versus biologic Ic disease—due to surface involvement or positive Douglas cytology—since the latter is associated with less favorable outcomes after FSS. In case of stage Ic disease and clear cell histology, there is increasing evidence of a poorer relapse-free survival compared to non-clear-cell histology. For that reason a fertility-sparing approach in this special patients collective should be indicated only after thorough discussion and informed consent of the affected patients with careful balancing of the risks and benefits. In any case, the treating gynecologic oncologist should be fully aware of his double role in treating the malignant disease as well as in providing oncofertility care to young EOC patients, by considering offering fertility-sparing alternatives when allowed so by tumor stage and histologic differentiation.


Panthera leo) and leopards (Panthera pardus), are financially valuable for ecotourism and trophy hunting operations on land also utilized for the production of other wildlife species for the same purpose. Predation of ungulates used for trophy hunting can create conflict with landholders and trade off thus exists between the value of lions and leopards and their impact on ungulate populations. Therefore productionist and conservation trade-offs are complexly graded and difficult to resolve. We investigated this with a risk-benefit analysis on a large private wildlife production area in Zimbabwe. Our model showed that lions result in substantial financial costs through predation on wild ungulates that may not be offset by profits from hunting them, whereas the returns from trophy hunting of leopards are projected to exceed the costs due to leopard predation. In the absence of additional income derived from photo-tourism the number of lions may need to be managed to minimize their impact. Lions drive important ecological processes, but there is a need to balance ecological and financial imperatives on wildlife ranches, community wildlife lands and other categories of multiple use land used for wildlife production. This will ensure the competitiveness of wildlife based land uses relative to alternatives. Our findings may thus be limited to conservancies, community land-use areas and commercial game ranches, which are expansive in Africa, and should not necessarily applied to areas where biodiversity conservation is the primary objective, even if hunting is allowed there.Large African predators, especially lions ( Where wildlife is utilized commercially, there can be conflicts among different forms of wildlife-use and between financial and conservation objectives Panthera leo) and leopards (Panthera pardus) have potential to generate significant returns from both photo-tourism and trophy hunting In Africa, wildlife occurs in a variety of scenarios, including At natural densities large African carnivores, especially lions, generally limit ungulate populations Lycaon pictus) In many African countries, large areas of land are designated hunting concessions or multiple-use zones, where trophy hunting is practised Giraffa camelopardalis), sable, and wildebeest (Connochaetes taurinus) remained relatively stable, and eland (Taurotragus oryx), kudu (Tragelaphus strepsiceros), waterbuck (Kobus ellipsiprymnus), zebra (Equus burchelli), impala (Aepyceros melampus) and warthog (Phacochoerus aethiopicus) numbers declined. Although most carnivores were present in SVC from 1996 the numbers of lions and spotted hyenas only started to increase markedly in 2006, whereas wild dogs increased to high numbers by 2003 and have declined slightly since then. Cheetah and leopard numbers appeared to be stable , yielding gross income of US$912,900 . During the same time 144 leopards were hunted (mean = 28.8 leopards year−1), generating a gross income of US$2,135,232 .From 2005 to 2009 seventeen male lions were hunted , combined with a 5% male lion harvest, resulted in a positive gross financial benefit for a population of up to 70 individuals. To balance out the predation effect of the current estimate of 160 lions, photo-tourism benefits per landowner would need to equal about US$ 13,000 year−1 , which needs to be evaluated against the objectives of the area. In our model we assumed that 5% of the lion population would be hunted as trophies. Such levels of harvest may be excessive for populations that have reached carrying capacity, for which quotas of about 3% are recommended Wildlife-based land uses are typically viable by virtue of the fact that multiple use values can be derived from wildlife, including trophy hunting, meat, skins and photo-tourism The situation in SVC is, however, not unique and thus our results apply to many other areas, such as large parts of Namibia Under a scenario of a managed lion population other large carnivores may temporally benefit. Both cheetahs 2 Savé Valley Conservancy (SVC) is located in southeast Zimbabwe in an area of semi-arid savanna typified by low and variable rainfall Loxodonta africana), African buffalo (Syncerus caffer) and sable (Hippotragus niger) contribute the bulk of the revenues generated from hunting, although ungulates are also extensively hunted The 3,440 kmAcinonyx jubatus) and spotted hyenas (Crocuta crocuta), with only leopards persisting in reasonable numbers Prior to the establishment of SVC, large predators were virtually eradicated from the area. Lions and wild dogs were eradicated, cheetahs (We developed a model to assess the net financial impacts of predators in SVC. The modelling process followed two key steps. First we built a biological model to estimate the numbers of prey species that would have been killed by the complement of predators estimated To determine the relative proportion of the various ungulate species in the large predator diet we used scat analysis data from SVC for leopards, wild dogs and spotted hyenas To estimate annual predation rates we determined the daily minimum food requirements for each predator species from literature The total numbers of ten most abundant small [50–10 k kg] and medium to large [>100 kg] herbivore prey species [excluding megaherbivores] killed per large predator species per year were estimated by combining the relative proportions of each herbivore in the diet, the biomass requirements of each large predator, and the numbers each prey species that an individual would likely have killed, multiplied by the annual population estimate of each large predator species see . This apWe estimated mortality by summing all quantifiable causes of ungulate mortality. The number of animals trophy hunted was determined from hunting returns. The number of animals found poached was recorded during a recent study, and adjusted upwards by 50% for under-recording of illegal off-takes −1. Since not all herbivores lost to predation or poaching would have been trophies, only the percentage of these taken by hunters [averaged from 2005–2009] was assumed to result in the loss of otherwise saleable trophies.Financial returns from trophy hunting were calculated as the trophy fee and the meat value of each animal hunted. The financial losses attributable to predation and poaching were calculated by summing the value lost in terms of meat, foregone trophy fees, and for buffalo and sable for foregone hunt-package fees. All trophy values, daily rates and minimum hunt durations were taken from the main hunting operator in the area Modelling is a compromise between the need for maximum realism and minimum complication in construction and design Lion hunting is an important income generator for Zimbabwe, and generally in African hunting concessions


Low birth weight has been associated with reduced hand grip strength, which is a marker of future physical function and disease risk. The aim of this study was to apply a twin pair approach, using both ‘individual’ data and ‘within-pair’ differences, to investigate the influence of birth weight on hand grip strength and whether this association may be mediated through fat free mass (FFM). Participants from the East Flanders Prospective Twin Survey were included if born without congenital abnormalities, birth weight >500 g and ≥22 weeks of gestation. Follow up in adulthood (age: 18–34 year), included anthropometric measures and hand grip . Birth weight was positively associated with hand grip strength  and FFM , adjusted for gestational age, sex and adult age. Using ‘within-pair’ analyses, the birth weight hand grip association was significant in DZ men only , which was attenuated following adjustment for FFM. Within-pair birth weight FFM associations were most pronounced in DZ men . Our ‘individual’ analyses show that higher birth weight is associated with greater adult hand grip strength, which is mediated through greater adult FFM. The ‘within-pair’ analyses confirm this observation and furthermore show that, particularly in men, genetic factors may in part explain this association, as birth weight differences in DZ men result in greater differences in adult strength and FFM. The association between low birth weight and increased disease risk in later life is well established Birth weight is used as a measure of prenatal growth, which is not only determined by the prenatal environment, but also by the foetus' genetic potential. As a consequence, associations between birth weight and adult disease may be due to a compromised environment in utero, or to genetic factors that affect both birth weight and adult disease risk. For example, it has been suggested that the previously observed association between low birth weight and increased risk of type 2 diabetes Twin studies provide a unique opportunity to disentangle the contribution of foetal growth restriction, maternal environment and genetic factors to the birth weight – adult disease association. This is because both members of a twin pair have the same gestational age and also share the same maternal exposures during pregnancy, such as maternal weight, nutrition and smoking status. However, each individual within a twin pair will have their own foetal-placental environment, which will contribute to differences in birth weight between the two twins. In monozygotic (MZ) twins, these differences in birth weight are solely due to the foetal-placental environment, whereas in dizygotic (DZ) twins differences in birth weight may also be due to genetic as well as environmental differences The aim of this study was therefore to investigate the influence of birth weight on hand grip strength in young adult twins. As it has been hypothesised that the association between birth weight and hand grip may be mediated through fat free mass The study was approved by the Ethics Committee of the Faculty of Medicine of the Katholieke Universiteit, Leuven, Belgium, and all participants gave written informed consent.The East Flanders Prospective Twin Survey (EFPTS) is a population-based twin survey that prospectively registered all twins born in the Belgian Province of East Flanders since 1964 We randomly contacted 803 pairs of all twins born between July 1964 and May 1982 during adulthood, when aged between 18 and 34 years, to participate in the Prenatal Programming Twin Study. Of those, 424 pairs  agreed to participate For the within-pair analyses, we excluded opposite-sex twin pairs (n = 43 twin pairs), as including pairs with discordant sex, where sex may be associated with both birth weight and hand grip, can lead to an overestimation of effect sizes. Pairs of which only one member participated in the study were also excluded, leaving a total of 326 same-sex twin pairs, 170 female pairs  and 156 male pairs .2 (m2). Fat free mass (kg) was estimated using bioelectrical impedance .Height was measured barefoot to the nearest 0.1 cm using a wall mounted stadiometer (Harpenden). Weight was measured to the nearest 0.1 kg on a balance scale  with the volunteer wearing light clothing. Body mass index (BMI) was calculated as weight (kg)/heightHand grip strength of the dominant hand was measured once using a Jamar hand grip dynamometer . Measurements were performed whilst standing, with the arm fully extended next to the body. Hand grip strength was measured on a single occasion after a demonstration and clear explanation by the research associate. Participants were able to familiarise themselves with the hand dynamometer prior to this measurement, without carrying out the test. Once the participants felt ready to carry out the test, they were verbally encouraged by the research associate to perform to their maximum ability.Mean and standard deviation were calculated for descriptive characteristics, for all individuals, for variables collected at birth as well as in adulthood. T-tests were used to investigate differences in means between MZ and DZ twins and between men and women.Twins were considered as individuals , to allow comparisons with other cohort studies, and as members of twin pairs .Linear regression was performed using mixed effect models, allowing for the fact that twins share more similarities than unrelated individuals, to investigate the associations between birth weight and adult hand grip strength. Similarly, the association between birth weight and adult fat free mass were analysed using linear regression mixed effect models.Regression models were adjusted for gestational age, adult age and where analysis was carried out in both men and women together the models were adjusted for sex. To investigate whether associations were independent of adult body size, models were additionally adjusted for adult height. Models investigating associations between birth weight and adult hand grip were also adjusted for adult fat free mass to examine whether associations were independent of fat free mass.Evidence for different effects in men and women were investigated by including an interaction term between birth weight and sex.difference in birth weight between the first and second born twin is modelled as the ‘exposure’ and the difference in hand grip between the first and second born twin is modelled as the ‘outcome’. This allows the influence of birth weight on hand grip strength to be estimated independently from shared maternal influences, such as maternal weight, nutrition and smoking status, and in MZ twins additionally from genetic factors.The Furthermore, comparing the effect size of the within-pair association of MZ twins to those of DZ provides information on the contribution of genetic factors to the birth weight – adult had grip strength relationship. For example, as MZ twin pairs share the same genes, any observed within-pair associations between birth weight and adult hand grip strength are more likely to be due to foetal-placental or environmental influences. Whereas larger effect sizes in DZ twin pairs, compared to MZ twins, would suggest that genetic factors contribute to the association between birth weight and adult hand grip strength.The associations between within-pair differences in exposure and outcome were analysed using linear regression. The first regression model was presented as unadjusted, or adjusted for sex where analysis was carried out in men and women together. The second regression model included adjustment for the within-pair difference in adult height, to investigate the influence of body size. Finally, hand grip models were also repeated adjusting for sex, within-pair difference in adult height and within-pair difference in adult fat-free mass, to investigate whether these associations were independent of body size and composition.All analyses were performed using Stata 9 .Female twins were just over 106 grams lighter at birth (p = 0.002) and had lower adult hand grip strength  than male twins Table 1.Birth weight was positively associated with hand grip strength  with a 1 kg increase in birth weight corresponding to a 2.6 kg increase in adult hand grip strength after adjustment for gestational age, adult age and sex Table 2.Additional adjustment for adult height attenuated the strength of association by more than half but remained significant . After adjustment for fat free mass the association was further attenuated and no longer significant Table 2.The strength of association between birth weight and adult hand grip strength in men was more than twice that of women, after adjustment for gestational age and adult age Table 2.We found that higher birth weight was consistently associated with increased fat free mass  and the strength of this association was more than halved after further adjustment for adult height . The association was twice as large in men  than in women  when adjusted for gestational age and adult age. Yet, the sex interaction was not significant . Adjustment for height attenuated the association, more so in women than in men .The association between within-pair difference in birth weight and the within-pair difference in adult hand grip strength was positive in direction but not significant , suggesting that, within a pair, the twin with the higher birth weight tended to have greater adult handgrip strength than their lighter sibling. When stratifying by sex, again there was a positive trend, but this was not statistically significant in either men or women. Further adjustment for intra-pair difference in either height or fat free mass completely abolished the association Table 3., When stratifying by zygosity status, to investigate the influence of genetic factors, we found a strong association between the within-pair difference in birth weight and the within-pair difference in adult hand grip in DZ men, while no association was observed in MZ men , Table 4Consistent with the cohort analyses, we observed a positive association between the within-pair difference in birth weight and the within-pair difference in fat free mass, indicating that the higher birth weight twin had more fat free mass in adult life compared to their lower birth weight twin. Again, this association was more pronounced in men than in women. Adjustment for height attenuated the association in men, and completely abolished the association in women.Positive associations between the within-pair difference in birth weight and the within-pair difference in adult fat free mass were observed in both DZ and MZ men, but this association was of a more than 4-fold greater magnitude in DZ men compared to MZ men Table 4 The opposite patterns of association were observed in women, as an association between the within-pair difference in birth weight and the within-pair difference in fat free mass, was only observed in MZ women but not in DZ women Table 4,Our cohort analyses that consider twins as individuals show that a higher birth weight is associated with greater adult hand grip strength, which is predominantly mediated through the association between birth weight and fat free mass. The overall trend from the twin pair analyses, which controls for shared factors, confirms the cohort observations and suggests that within a pair the heavier twin at birth has greater hand grip strength in adult life than their lighter sibling. Uniquely, the twin pair analysis identified that the association between birth weight and hand grip strength was stronger in DZ than in MZ men, suggesting a potential influence of genetic factors, acting on both on birth weight and adult hand grip in men.The positive associations between birth weight and adult hand grip strength are consistent with those observed in singletons and show similar effect sizes Our findings suggest different patterns in the association and mediation between birth weight, and hand grip and fat free mass for men and women. In women the association between birth weight and hand grip strength was completely abolished after adjustment for adult height or fat free mass. In addition the association between birth weight and adult fat free mass in women was substantially reduced after adjustment for adult height. In men, however, while the association with hand grip is attenuated after adjustment for adult height or fat free mass, there remains a residual effect of birth weight on adult fat free mass independent of adult height. This suggests that the association between birth weight and grip strength is mediated through both adult height and fat free mass in women, whereas in men the association is largely mediated through fat free mass.2) in boys The gender-specific association between birth weight and adult fat free mass is consistent with the results observed in a study in children that used the more sensitive four-compartment measurement of body composition. In this study, birth weight was associated with height in both boys and girls, but birth weight was only associated with fat free mass (measured as Fat Free Mass Index = FFM/heightA potential explanation for the difference observed in the birth weight fat free mass association observed in this study, which was only independent of height in men, is that adult muscle mass distribution is known to vary between men and women both birth weight and adult hand grip. A similar pattern is observed for fat free mass; i.e. in the association was more pronounced in DZ men than in MZ men, again suggesting that while in utero growth restriction influences adult fat free mass, there are additional genetic factors acting on both birth weight and later fat free mass.A novel finding in this study was the significant association, using within-pair analysis, between birth weight and adult hand grip strength in DZ but not in MZ men. This suggests that, at least in men, the association between birth weight and adult grip strength may in part be explained by genetic influences that act on difference in birth weight between the first and second born twin. The range of such within-pair differences are typically much smaller than the range of birth weights observed for individuals using conventional cohort analysis, making it harder to detect associations, so it is possible that the within-pair analysis was underpowered. Furthermore, it should be noted that this present study used bio-electrical impedance to estimate fat-free mass, which may not be as accurate as other laboratory based measures such as duel x-ray absorbtometry, densitometry or deuterium dilution, although many of these more detailed measures are less feasible in larger studies. It should also be noted that we did not measure physical activity, which can influence both fat-free mass and physical performance. Therefore, it is possible that the associations between birth weight and fat-free mass may be mediated by physical activity levels. Finally, we acknowledge that twins may not be representative of singletons, as twins tend to be born both earlier and lighter. Therefore, the results should be interpreted with caution. However, we should highlight that our twin sample is randomly drawn from a prospective, population-based cohort with detailed information collected at birth and with objective measures of muscle strength and body composition in young adulthood.There are however some limitations to this present study. While the twin pair analysis has a number of advantages for disentangling genetic and in utero effects, it has typically less statistical power to detect associations as compared to the individual cohort analyses. Firstly, because using twin pairs as the unit of analysis effectively halves the sample size, secondly the within-pair analysis uses the Future large scale twin studies would be particularly useful to investigate the developmental origins of physical performance and body composition further. Elucidating the influences of shared maternal factors, both in terms of biological influences such as maternal weight and nutrition status, as well as confounding from socio-economic factors, is one of the key issues when investigating the developmental origins of health and disease. Therefore, future twin studies with detailed maternal measures of these variables would be particularly useful.Future genome wide association studies may identify common genetic variants influencing birth weight, muscle strength and body composition. For preventive purposes, it would be particularly interesting to understand whether the associations between these potential gene variants with future body composition and physical function, including muscle strength, are modified by lifestyle factors, such as increased physical activity.In summary, our findings suggest that birth weight is positively associated with adult hand grip strength, which appears to be mediated predominantly via influences of birth weight on fat free mass. Furthermore, in men these associations may, at least in part, be influenced by genetic factors that act on both birth weight and later hand grip strength and fat free mass. These findings suggest that a compromised early life, due to either growth restriction in utero or other maternal factors, may be important for both hand grip strength and fat free mass in adulthood, which has implications for both physical function and disease risk in later life.


This review provides an introduction to mass spectrometry based proteomics and discusses several proteomics approaches that are relevant in understanding the pathophysiology of fibrotic disorders and the approaches that are frequently used in biomarker discovery. What are the changes in protein levels that occur during or because of fibrosis? What are the signaling mechanisms associated with fibrosis? Mass spectrometry based proteomics has been successfully used to study many diseases in order to identify potential biomarkers and/or understand pathogenesis, however, there haven't been many studies devoted to understanding proteins and their post-translational modifications (PTM) as connected with fibrosis in different tissues.One of the reasons for this is probably technological - up to recent years the majority of proteomics techniques were not sensitive enough to identify and/or quantify minor changes in protein expression levels. The analysis of PTMs, on the other hand, is challenging because those modifications are usually in low abundance, complex or transient in nature. This overview will discuss mass spectrometry based proteomics techniques for protein characterization and point out those studies that have successfully used the approaches to study fibrosis and to search for potential biomarkers in fibrotic tissues or patients plasma.Proteomics strives to characterize proteins and their role in complex biological systems. It is only one of the components of -omics approaches which - when fully applied to a biological problem - will generate a composite picture of biological processes. Mass spectrometry based proteomics is the first step to study the changes that proteins undergo as a consequence of fibrotic disorder, whether the changes are in protein levels , temporal (following the development of a disorder), or on the post-translational level (e.g. phosphorylation) as compared to the control, healthy tissue or plasma. The final goal of many proteomics studies is the discovery of a biomarker protein or a protein panel that enables detection of disease, can monitor and measure the treatment progress and could also predict development of the disease. Majority of studies so far have used patient plasma to elucidate the mechanisms of fibrosis or find its putative biomarkers as compared to relatively few studies that use either tissues after biopsy or animal models/cell lines.Mass spectrometry is routinely used to identify proteins, quantify their levels (relative or absolute) and to characterize protein post-translational modifications. Some useful reviews on the application of different mass spectrometry based approaches were written by Yates et al. ,2 and CoThe advancements in the mass spectrometric technology, such as in instrument designs  and mass analyzers (orbitrap) have significantly improved the sensitivity and mass accuracy. Nevertheless, sample preparation and fractionation are very important especially when dealing with scarce clinical samples to be able to characterize high numbers of proteins and their post-translational modifications. Dealing with tissue samples or plasma poses additional challenges because of sample stability, solubility and abundance issues. Proteins in tissues are usually difficult to solubilize , whereas plasma contains abundant proteins  that can easily mask and thus prevent the identification of lower abundant proteins . With the depletion of most abundant plasma proteins (up to 20), this approach then enables mass-spectrometry based proteomics to dig deeper into plasma proteome in order to identify low abundant proteins and their possible modifications.The final goal of many proteomics experiments is to completely characterize a proteome: protein's identification, location, function, interacting partners, post-translational modifications (PTMs) and in many cases also the turnover rate of protein production/degradation or the rate of formation for a particular PTM. We would also like to know what are the occupancies for a particular PTM (e.g. what percentage of a given site in protein is phosphorylated). Some recent reviews on PTMs are the following: phosphorylation -6, acetyMass spectrometry is an indispensible tool for relative protein quantitation which is usually applied to discovery of clinical biomarkers and mostly uses large-scale proteomics experiments. The first step in a typical large scale proteomics experiment is a separation technique on either the protein e.g. 1D gel) or peptide level (e.g. SCX - strong cation exchange chromatography), followed by analysis of each of the individual fractions with liquid chromatography coupled to tandem mass spectrometry system (LC-MS/MS). The liquid chromatography is mostly reverse phase based with the flows in nanoliter range. Usually, 1D gel separation on protein level followed by an LC-MS/MS analysis gives a higher number of identified proteins because the first dimension separation is on the protein level as compared to SCX which separates peptides on the basis of charge. A summary of proteomics approaches using mass spectrometry is depicted in Figure D gel or The analysis of PTMs usually involves enrichment which is based on an antibody pull-down or it takes advantage of some physicochemical property of the PTM to selectively target proteins or peptides with that PTM. For example, tyrosine (Tyr) phosphorylated proteins (or peptides) are easily enriched by using a specific anti-pTyr antibody. On the other hand, serine or threonine phosphorylated proteins or peptides are enriched by using immobilized-metal affinity chromatography (IMAC) and metal-oxide affinity chromatography (e.g. TiO2 based). After the enrichment, the mass spectrometric analysis can pose some difficulties as well. A lot of PTMs are labile modifications that can easily fragment under collisionally induced dissociation in mass spectrometer .There are several complementary approaches for relative protein quantification, such as DIGE (2D flourescence difference gel electrophoresis) , iTRAQ . In the quantitative approach (DIGE), two protein samples are labeled with fluorescent dyes and when combined and separated on a 2D gel can be visualized by measuring the fluorescence showing whether a protein is up- or down regulated and for how much. The identification of individual spots of interest is carried out by mass spectrometry. The gels are usually scaled up (pick gel), stained with Coomassie blue, excised and digested with trypsin before being analyzed on a MALDI TOF/TOF or electrospray type of an instrument. Since fluorescence can be very sensitive, it is sometimes difficult to identify the proteins in spots by mass spectrometry which is a couple of orders less sensitive. Another drawback of this approach is that sometimes the individual spots may provide several identifications with similar proteins co-eluting.As a complementary technique to DIGE, iTRAQ has recently gained a lot of popularity in the protein biomarker applications because of the ability to multiplex several samples in one run (up to 8) and because the number of identified proteins are usually higher.Proteomic analysis of fibrotic tissue is further complicated with the limited ability of most common techniques to solubilize proteins in a complicated matrix. Usually, the number of identified proteins from a tissue sample is smaller as compared to a cell lysate. Most of the biologically important proteins  are of lower abundance, thus it is essential to identify as many proteins as possible. The other drawback associated with fibrotic tissue analysis is limitation with the quantitation techniques because some of them due to the nature of labeling cannot be applied (e.g. SILAC vs. iTRAQ). Plasma samples, on the other hand, pose challenges as well, mainly due to high abundant proteins that are usually depleted to enable identification of less abundant proteins in the sample.Liver fibrosis has been relatively well studied using available proteomics tools and there are detailed reviews that critically discuss the use of proteomic approaches in hepatic fibrosis, including the search for putative biomarkers ,15. In aRenal fibrosis and complex proteomics associated with chronic kidney disease has been reviewed in a recent paper by Klein et al. . Two othIdiopathic pulmonary fibrosis (IPF) - which is a disease of unknown etiology and is connected to progressive lung fibrosis - is the most studied condition related to lung fibroproliferative disorders. Kinulla et al.  reviewedAmong interesting proteins revealed and further studied by proteomics approaches, apolipoprotein A-I may be useful in therapeutic strategies after it was recently shown that its levels are decreased in IPT lungs compared to normal tissue animal models . SimilarCastro et al.  criticalScambi et al.  investigMass spectrometry based proteomics is a powerful tool to identify proteins and their posttranslational modifications. When applying these techniques to fibrotic disorders, the sample preparation is critical - with biological fluids it is possible to remove the most abundant proteins by immunodepletion. Liver fibrosis has been relatively well studied using available proteomics tools and there are several possible biomarkers that were identified by using proteomics approaches.The current state of technology definitely brings along a lot of opportunities, and challenges as well, especially with the data analysis and making sense of the protein lists.DIGE: 2-D Fluorescence Difference Gel Electrophoresis; PTM: post-translational modification; LC-MS/MS: liquid chromatography coupled to tandem mass spectrometry; iTRAQ: isobaric tag for relative and absolute quantitation; SILAC: stable isotope labeling with amino acids in cell culture.The author declares that they have no competing interests.


Environmental and genetic interventions extend health span in a range of organisms by triggering changes in different specific but complementary pathways. We investigated the gene expression changes that occur across species when health span is extended via different interventions. To perform this comparison using heterogeneous datasets from different measurement platforms and organisms, we developed a novel non-parametric methodology that can detect statistical significance of overlaps in ranked lists of genes, and estimate the number of genes with a common expression profile. By comparing genetic and environmental interventions that consistently lead to increased health span in invertebrates and vertebrates we built a conserved health span signature and described how such a signature depends on tissue type. Furthermore, we examined the relationship between calorie restriction and resveratrol administration and for the first time, identified common gene and pathway changes in calorie restriction and resveratrol in both invertebrates and mammals. Our approach can thus be used to explore and better define the relationships between highly complex biological phenomena, in this case those that affect the health and longevity. Aging is a very complex phenotype: it can assume different forms in different species, individuals, and tissues, and its mechanisms are multiple, complex and stochastic in nature. Several interventions that extend life span in many organisms have been identified, including environmental factors, genetic manipulations and drugs. It is thought that these diverse interventions share some common pathways through which they extend life span.Drosophila melanogaster and its mammalian homolog Sirt1, has been extensively studied in the field because of its role in reducing age-related pathologies in a wide range of organisms D. melanoets from . GCRMA [ets from  was usedk* genes in the intersection of two lists generated by randomly choosing two sets of m genes out of a total of N was computed as:http://www.physics.brown.edu/physics/researchpages/ibns/listcomp.htm.The probability of observing at least ijm = maxl,m)=1…N( (lmk) − ijk as metrics, where N is the total number of experimental conditions to be compared, and ikj is the number of genes in the intersection when comparing the i-th experimental condition to the j-th experimental condition.Hierarchical clustering was performed using a complete linkage algorithm and using Probesets identification numbers in the fruit fly and mouse microarrays were converted to the corresponding gene symbol and matched across species using the HomoloGene database from NCBI.


Stroke patients recover more effectively when they are rehabilitated with bimanual movement rather than with unimanual movement; however, it remains unclear why bimanual movement is more effective for stroke recovery. Using a computational model of stroke recovery, this study suggests that bimanual movement facilitates the reorganization of a damaged motor cortex because this movement induces rotations in the preferred directions (PDs) of motor cortex neurons. Although the tuning curves of these neurons differ during unimanual and bimanual movement, changes in PD, but not changes in modulation depth, facilitate such reorganization. In addition, this reorganization was facilitated only when encoding PDs are rotated, but decoding PDs are not rotated. Bimanual movement facilitates reorganization because this movement changes neural activities through inter-hemispheric inhibition without changing cortical-spinal-muscle connections. Furthermore, stronger inter-hemispheric inhibition between motor cortices results in more effective reorganization. Thus, this study suggests that bimanual movement is effective for stroke rehabilitation because this movement rotates the encoding PDs of motor cortex neurons. One of the challenges of rehabilitation research is to elucidate efficient method of promoting the functional recovery of upper limb movement in stroke patients. In neuroscience, a related challenge is determining the neural mechanisms of such functional recovery. Although stroke patients tend to recover lower limb movement after therapeutic intervention, the majority of these patients (65%) do not regain full movement of their upper limbs A recent computational study suggested that constraint-induced therapy is effective because it leads to the reorganization of a damaged region in the motor cortex based on supervised and unsupervised learning Although previous computational studies investigated the unimanual movements of stroke patients, individuals often move their arms bimanually. Bimanual movement is effective for the recovery of paretic arm movement Neural activities in the motor cortex differ during unimanual and bimanual movement Using a computational model of stroke rehabilitation This study investigated how bimanual movement affected stroke recovery by modeling the different PDs and modulation depths during unimanual and bimanual movement. During bimanual movements, we invesitigated only bimanual parallel movements in which the right and left arms move in the same directions, because the neural activities during these parallel bimanual movements have been investigated previously. We describe the definitions used in the model in detail in the Importance of encoding PD rotations for reorganization, we consider changes of modulation depth, i.e., Initially, in simulating bimanual rehabilitation, we assume that changes occur only in the PDs. These changes are referred to as PD rotations for the remainder of this manuscript. Because PDs are rotated pseudo-randomly in bimanual movement, we modeled these rotations asImportance of encoding PD rotations for reorganization, the modeling of rotations in decoding PDs is described.Decoding PDs are used to calculate the population vector (PV) as follows:We can model the upper limb movements of stroke patients using a PV by removing a fraction of the model neurons After a neuronal lesion, we investigated whether reorganization in the damaged motor cortex can be facilitated by rotations of the encoding PDs. When the encoding PDs are not rotated  and 60 (45+15) degrees. However, in this case, the norm of the PV is smaller than that of the well-equalized population; this reorganization is observed when the encoding PDs are strongly rotated. Furthermore, in well-equalized populations, more neurons are activated in generating PVs oriented at 45 degrees, and movement precision is better than that observed in the impaired populations observed after unimanual rehabilitation. In other words, in unimanual rehabilitation, the reorganization stops at one of the local solutions for which the angular error is decreased to some extent, but movement speed does not recover. When non-quenched  random variables are added to each rehabilitation (learning) step, these variables act as search noise and cause reorganization, thus avoiding trapping in local solutions This study investigated the influence of quenched  rotations of encoding PDs on reorganization, but annealed  rotations can also be considered. However, we observed no significant reorganization when the rotations were annealed (data not shown). By contrast, when we added annealed noise to each learning step see Methods,This study assumed the following conditions: subjects move their arms towards one of eight radially distributed targets at angles of Based on these neural activities, the neural population generates a population vector The PV can be used to model the reaching movements of stroke patients by depleting a fraction of the model neurons After the neuron depletion we modeled the reorganization process of a damaged motor cortex as the optimization process of the following cost function:We assumed that bimanual movement induces rotations of the PDs only during encoding and not in decoding


Targeted Benefits, and as Co-Benefits/Costs (in Marxan's cost function); we also compared these approaches with a Hybrid approach . For this analysis, the Co-Benefit/Cost approach yielded a less costly reserve network than the Hybrid approach (1.6% cheaper). Including timber harvest as an opportunity cost in the cost function resulted in a reserve network that achieved targets equivalently, but at 15% lower total cost. We found counter-intuitive results for conservation: conservation-compatible services  were positively correlated with each other and biodiversity, whereas the conservation-incompatible service (timber) was negatively correlated with all other networks. Our findings suggest that including ecosystem services within a conservation plan may be most cost-effective when they are represented as substitutable co-benefits/costs, rather than as targeted benefits. By explicitly valuing the costs and benefits associated with services, we may be able to achieve meaningful biodiversity conservation at lower cost and with greater co-benefits.There is growing support for characterizing ecosystem services in order to link conservation and human well-being. However, few studies have explicitly included ecosystem services within systematic conservation planning, and those that have follow two fundamentally different approaches: ecosystem services as intrinsically-important targeted benefits vs. substitutable co-benefits. We present a first comparison of these two approaches in a case study in the Central Interior of British Columbia. We calculated and mapped economic values for carbon storage, timber production, and recreational angling using a geographical information system (GIS). These ‘marginal’ values represent the difference in service-provision between conservation and managed forestry as land uses. We compared two approaches to including ecosystem services in the site-selection software Marxan: as  Although the area of land protected has expanded considerably across the planet Ecosystem-based approaches to management are rapidly gaining momentum, with governments across the world requiring the simultaneous consideration of ecological sustainability and the flow of multiple benefits to people (ecosystem services) from ecosystems Biodiversity conservation is often a catalyst for ecosystem-based approaches to management. Yet, conservation planning lacks an established means to measure the full extent of costs and benefits associated with alternative conservation plans, for people. Because ecosystem services can be important co-benefits or opportunity costs of conservation, there has been considerable interest in ecosystem services in planning As the migration toward ecosystem services-based approaches for management increases, two key questions must be answered: (1) how well do biodiversity and ecosystem services correlate across space? And, given imperfect correlations, (2) how can we use existing planning tools to most effectively prioritize for a range of management considerations within a particular landscape and on a constrained budget? A central challenge in coupling ecosystem services research and conservation planning is that many efforts to value services To integrate ecosystem services into conservation planning, it is helpful to develop frameworks for marginal valuation compatible with the prevailing tools of reserve-design, such as Marxan intrinsically important within a framework of cost-minimization, whereas the Co-Benefit/Cost approach includes them as substitutable in a framework of net-benefit-maximization—a critical philosophical difference  will frequently be at odds with biodiversity conservation. But even in such cases of incompatibility, there may be great gains in conservation efficiency by including these services in conservation planning as opportunity costs We present spatially explicit, marginal economic values of three ecosystem services  and integrate them in the Targeted Benefit, Co-Benefit/Cost, and Hybrid approaches to answer the two key questions posed above. Carbon storage refers to the carbon stored in above and below ground biomass as well as soil We chose to investigate these three services based on an informal survey of the NCC Central Interior team of experts, and in consideration of available data. All three services were chosen based on the demonstration of changes in value due to timber harvesting (the greatest threat to conservation in the area) and their importance to local beneficiaries. The analysis consisted of two primary objectives: (1) obtaining an economic valuation of ecosystem services and mapping these services with biodiversity features and (2), the inclusion of these values for a set of Marxan analyses, in part to compare the efficiency of the Targeted Benefit, Co-Benefit/Cost, and Hybrid approaches. The goal of this work is to guide conservation thinking in the study region regarding the extent to which recreational angling, timber harvest, and carbon storage offer opportunities or obstacles for conservation, the places where these opportunities/obstacles arise most strongly, and the means by which systematic conservation planning might account for these services.The estimated net present value of changes in ecosystem services associated with a difference in management between timber harvesting and conservation are highly variable within and between services . Values It is unsurprising that timber harvest values dominate the other two services modeled, given that managed forestry is the prevailing land use/land cover in the region. A small but notable fraction of the landscape is characterized by negative (red) or negligible (light yellow) timber values, meaning that harvest-operation costs are projected to outweigh benefits . Such loThe reserve network for biodiversity features is much patchier than the networks for ecosystem services , as mighTimber production represents the greatest opportunity cost to conservation in the study area. The hotspots chosen for timber are primarily in the center of the study area, close to roads as reflecting ease of transportation. These areas do not overlap with the hotspots for carbon storage. Therefore, opportunity costs related to timber production would be relatively low in the areas chosen for a carbon storage reserve network, thereby increasing the likelihood of conservation.Conservation-compatible services (angling and carbon storage) were positively correlated in space with biodiversity, and all these were negatively correlated spatially with timber harvest . The corOur representation of ecosystem service values provides a novel and powerful approach for investigating general relationships between pairs of benefits, including ecosystem services and biodiversity, and between approaches to integrating services in conservation planning. Due to the nascent stage of research on the production and impacts to ecosystem services In our study area, treating ecosystem services as Targeted Benefits generally yielded more spatially cohesive but costlier reserve networks than treating services as co-benefits or costs (our Co-Benefit/Cost or Hybrid approaches). While this may be due partly to the particular parameters and details of our study, there is reason to believe that the efficiency gains of the co-benefit/cost approach are general, if one assumes that the ecosystem-service valuations and their inclusion alongside other economic costs is accurate. The Co-Benefit/Cost approach represents the importance of ecosystem-service values to total costs/benefits directly, whereas the Feature approach does so through a suite of parameters ). The purpose—and the advantage—of the Co-Benefit/Cost approach is therefore efficiency. Accordingly, if services have associated economic values , and those values are well known, including those values in the cost surface will be the simplest means of ensuring the services are given their due weight.Efficiency is not all-important, however, so even if economic valuations are trustworthy, the choice between the two approaches in any given context should depend largely on the nature of the benefits and values at stake. Integrating service values into the cost surface effectively treats the benefits at stake as substitutable costs or benefits that might serve instrumental roles towards conservation. Treating services as benefits for which targets should be set (and achieved) effectively treats the benefits as inherently important and not perfectly substitutable with other benefits. Whereas the former is appropriate if the principal values at stake are preference-based, as with market values, the latter is much more appropriate if the values at stake include principles and virtues Our study revealed a mild good-news story for conservation, in the positive correlations between conservation-compatible services and biodiversity and the negative correlations of the service considered incompatible with conservation in this this case (timber production). The spatial incongruence between carbon storage and timber production networks is an unexpected positive outcome for conservation. One might expect that areas of high timber production value would align with high carbon storage, but they did not. Many areas with high carbon storage values are characterized by variable slope, which results in high harvesting costs and thus low timber production values. Although it is important to confirm this result with more sophisticated forestry modeling, this incongruence between timber harvest values and conservation might assist conservation of biodiversity and compatible services based on relatively low opportunity costs of timber harvest.The relatively weak and sometimes negative correlations between ecosystem services and biodiversity shown here echoes past research on ecosystem services in conservation planning Tradeoffs are expected between certain services and biodiversity for a variety of reasons Ecosystem service research and, particularly, spatially explicit approaches to doing so are often restricted by available data. Similarly, our analysis is limited somewhat by inconsistencies in the original data. In particular, the model of timber production was based on timber supply reviews, which are reports conducted for individual timber supply areas (TSAs) in British Columbia All of the networks that used a Co-Benefit/Cost or Hybrid approach  consisted of many small patches that are likely not realistically implementable as conservation areas. As such, further experimentation with Marxan's parameters and connectivity analysis is needed to realize the potential of this novel approach.potential, not realized benefits, but the gains of including these values depend on the realization of these values. Benefits could be realized if the conservation agent  were to receive any of the following: funds for carbon offsets for carbon stored on lands that would have been logged; assistance (or less opposition) from the forestry industry in exchange for bypassing conservation protection of high timber-value areas; assistance or funding from recreational angling groups for conserving areas of importance to them; or other incentives associated with new policies to internalize costs and benefits of ecosystem services. Institutional changes such as the creation of carbon storage credits and/or markets are critically enabling here. Without such policies and institutions for other ecosystem services, ecosystem-service contributions to better land-use and management decisions will be limited and dependent on individual good will and leadership.The values associated with these ecosystem services are Our study both confirms previous results and offers a significant advance to the incorporation of ecosystem services into conservation planning. Our case study reveals weak positive associations between conservation-compatible services  and biodiversity, and weak negative associations of these with conservation-incompatible timber harvest. Further, it demonstrates the considerable efficiency gains of including more sophisticated cost (and co-benefit) data into planning: including services in conservation planning analyses will appropriately skew prioritization towards areas with lower costs or greater co-benefits. Our central advance is a demonstration of contrasting approaches to including services in planning: treating services as targeted benefits vs. as co-benefits/costs. In our study, the two approaches—which differ in their applicability, depending on the service—performed roughly similarly, with moderate efficiency gains for the co-benefit/cost approach.The challenge of modeling ecosystem services is immense, but so is the opportunity: conservation projects designed and implemented effectively can benefit people and ’nature’; and rigorous analysis of ecosystem-service benefits could be instrumental in inspiring the institutional changes needed to internalize these important values in decision-making.2  static assessment of current carbon storage with a model representing how carbon storage varies across time in a given forest landscape. For the former, we used publicly available data from the World Resources Institute containing information about carbon storage in soil, as well as above and below ground vegetation This model was used to determine the change in carbon storage in two hypothetical BC Interior forest landscapes that differed only by their fire disturbance and managed harvest cycles. The landscapes had fire disturbance cycles of 500 and 750 years and harvesting cycles of 100 and 120 years respectively. To calculate carbon loss we adopted findings from Kurz et al. Our economic values for carbon storage reflect avoided social damages associated with climate change. We valued carbon storage at $8.46 (CDN) per ton of carbon dioxide using the mid-price average of three carbon trading markets: the Chicago Climate Exchange, the New South Wales and the EU Emissions Trading scheme on March 19th, 2008. We followed the methods and assumptions outlined by Naidoo and Ricketts The coarse resolution of the data hides variations within each cell; thus carbon values represent averages across individual stands at different ages/stages, with a distribution of stages resulting from the historic management regime. We assumed that each cell has been managed according to a sustainable yield model for harvest rotations specified by its particular TSA, such that for cells that are managed with a 40-year rotation approximately 1/40th of the harvestable area in each cell was harvested each year for the past 40 years. Accordingly, we may slightly overestimate or underestimate the change in carbon storage in any cell based on whether harvest was more or less recent than half of the listed rotation length. Subject to these assumptions, our value represents the net present value (NPV) of social damage avoided by the difference in carbon storage associated with timber harvesting/conservation.Timber production is measured here as an opportunity cost of conservation, with the difference between the two land-use scenarios being 100% of the net value of timber harvest. NCC does not intend to allow regular timber harvest within reserves.We calculated NPVs for timber production over a 1000-year timeframe with a discount rate of 4%, assuming a constant ratio of benefits to costs, based on expert opinion and forestry economics literature Costs consisted of harvesting costs, cost of transportation to the closest processing facility, and the costs of replanting (silviculture costs). These costs were based on slope, distance and biogeoclimatic zone, respectively, and were derived from previous merchantability assessments in the province We assumed that timber harvesting and associated activities will have an adverse effect on recreational angling values through an increase in sedimentation We determined the value of recreational angling in the study area, and how much it may be impacted by timber harvesting activities, using data from an angler effort model that predicts how much actual angler effort (measured in days spent fishing) can be supported by a particular lake given its productivity, distance from major population centers, and accessibility by roads. Parkinson et al. Using the Ministry of Environment's Fisheries Sensitive Watershed database, we assigned relative sensitivity scores (from 0 to 1.0) to third-order watersheds in the study area based on equal weighting of six characteristics: soil type, density of alluvial streams, lake buffering capacity, amount of forest cover, annual precipitation, and slope For simplicity in the absence of other understanding, we assumed a linear 1∶1 relation between sensitivity to timber harvesting and change in economic values of recreational angling. Thus we combined the sensitivity score of each watershed with its recreational angling value, based on amount of effort, to derive a final value of the expected difference in recreational angling values between conservation and timber harvest scenarios .Biodiversity features were divided into coarse and fine filter features such as old growth forest ecosystems and rare plant species, respectively. The fine filter data consist of over 75 plant species and 100 animal species . Animal species were selected based on their designation as threatened on provincial, national, and international lists, as well as other more expert-informed subjective characteristics such as whether the species is endemic, regionally important or especially vulnerable to change. Data used to represent these features came from a variety of sources including the BC Conservation Data Centre, the BC Ministry of Environment, the Canadian Wildlife Service and Ducks Unlimited.The data represent terrestrial ecological systems, as defined by the NatureServe classification system We used the site selection program Marxan (version 2.0.2) to select reserve networks Our results focused on the “best” and “summed solutions” outputs. The “best” solution from each scenario is the reserve network that has the lowest objective function score and meets all targets We did not incorporate variable threats (probability of habitat loss) but rather assumed that all areas not protected would eventually be subject to degradation (the ‘scorched-Earth’ approach). This was in line with the approach of the NCC and their ecoregional assessment. While we see great value in incorporating a threats analysis into conservation planning, there was no available defensible threats analysis for this study region.Wherever possible, we maintained parameter values as closely as possible across scenarios. In some cases, this may seem odd , but it is important for two reasons. First, the context here is of conservation, and if ecosystem services are to be helpful for biodiversity protection, the management needs and costs will parallel those of biodiversity, to a degree. Second, maintaining consistency of parameters across scenarios is key for maintaining comparability across scenarios.Ecosystem services have generally been included in conservation assessments as benefits for which particular targets are desired We ran separate scenarios in Marxan that included carbon storage, recreational angling and timber production ecosystem service values in the suitability index (using the transformation explained below). By associating ecosystem service values as co-benefits (for angling and carbon) or costs (for timber) with each planning unit, we effectively increase or decrease the cost of including the planning unit in the reserve. Despite calls to do so, researchers have rarely included socio-economic values in choosing areas for conservation ; this novel Benefit/Cost approach offers a simple means for conservation planning to better reflect implications for human well-being.To integrate ecosystem-service values into the SI, we had to convert the SI road index scores into dollar values reflecting fee-simple acquisition costs. NGOs including NCC also ‘acquire’ land through conservation easements For the conversion to dollar values we used a four-part linear transformation based on land prices (acquisition costs) in the study area, assuming that higher SI scores (higher road density or proximity to roads) correlate with higher land prices due to urbanization. Because NGOs also receive land donations, the market price may be an overestimate of acquisition costs, possibly rendering our transformation to market prices somewhat conservative. But it is impossible to account for the unpredictability and site-specificity of donations at an ecoregional scale. For greater detail on the SI and our transformation, please see Appendix C .We then added timber production values and/or subtracted recreational angling and carbon storage values from the transformed SI. We thereby assume that ecosystem services increase (in the case of timber production) and/or decrease  the costs or difficulty of conservation. For example, an area with high carbon storage values may be more easily conserved as some of the costs of conservation might be recouped through future fiscal returns via carbon credits. In a similar way, an area with high timber production values may be found to have opportunity costs that render conservation socially unacceptable.ad hoc calculations of total costs, to ensure fair comparisons. First we identified the areas in the “best” reserve network for biodiversity, recreational angling and carbon storage benefits together (our Targeted approach). We then determined the cost of this reserve network inclusive of ecosystem service values —although the cost surface of Marxan in this case had not included the service values. We compared this summed cost to the cost of a reserve network that protected biodiversity but included recreational angling, carbon storage and/or timber production values within the cost surface (our Co-Benefit/Cost approach).We compared the efficiency of this Co-Benefit/Cost approach to the Targeted Benefit approach by comparing the cost of the “best” solutions in the two approaches In order to assess the spatial correlation between “summed solutions” for each scenario we calculated Pearson's correlation coefficients for each pair of scenarios . These vText S1Three appendices providing additional details on methods. Appendix A ; Appendix B (details of Marxan scenarios); and Appendix C (suitability index transformation).(DOCX)Click here for additional data file.


Lung cancer is the most common cancer in the world. Despite modern advancements in surgeries, chemotherapies, and radiotherapies over the past few years, lung cancer still remains a very difficult disease to treat. This has left the death rate from lung cancer victims largely unchanged throughout the past few decades. A key cause for the high mortality rate is the drug resistance that builds up for patients being currently treated with the chemotherapeutic agents. Although certain chemotherapeutic agents may initially effectively treat lung cancer patients, there is a high probability that there will be a reoccurrence of the cancer after the patient develops resistance to the drug. Erlotinib, the epidermal growth factor receptor (EGFR)-targeting tyrosine kinase inhibitor, has been approved for localized as well as metastatic non-small cell lung cancer where it seems to be more effective in patients with EGFR mutations. Resistance to erlotinib is a common observation in clinics and this review details our current knowledge on the subject. We discuss the causes of such resistance as well as innovative research to overcome it. Evidently, new chemotherapy strategies are desperately needed in order to better treat lung cancer patients. Current research is investigating alternative treatment plans to enhance the chemotherapy that is already offered. Better insight into the molecular mechanisms behind combination therapy pathways and even single molecular pathways may help improve the efficacy of the current treatment options. Lung cancer is a leading cause of cancer-related deaths and has one of the lowest survival rates among all cancers. In 2008, lung cancer was found to be the most commonly diagnosed cancer as well as the primary cause of cancer-related mortality for males worldwide and the second leading cause of cancer-related deaths for women  and NSCLC. SCLC is a type of lung cancer that is characterized by short cell doubling time, rapid, aggressive progression, and early occurrence of blood-borne and lymph metastasis , a platinum-based therapeutic treatment, is usually utilized to treat NSCLC and has been shown to effectively improve patient survival  is a tyrosine kinase that contributes to the regulation of cellular homeostasis Cohen, . It is aStudies have identified some EGFR mutations in the tyrosine kinase domain in NSCLC patients which may help predict therapeutic results with tyrosine kinase inhibitors (TKIs), erlotinib, and gefitinib  activation and amplification and resistance to TKIs  is associated with increased cell invasion, migration, and proliferation and loss of cell adhesion proteins in numerous cancer cell lines and linked to the progression of cancer malignancies . Emerging studies have linked epigenetic changes, including DNA methylation at CpG islands, to the development of drug resistance to many anti-cancer drugs  inhibitor trichostatin A (TSA) to enhance the effects of erlotinib  and phosphatidylinositol-3-kinase (PI3K) pathways, affects signal transduction linked to angiogenesis, apoptosis, and cell proliferation  inhibitors, thalidomide, biological vaccines, and small molecular weight inhibitors directed at receptor protein tyrosine kinases. Many of these have been tested in phase II clinical trials for success in overcoming drug resistance  are small regulatory RNA molecules that post-transcriptionally regulate gene expression by targeting messenger RNAs (mRNAs; Croce and Calin, Cancer research has progressed rapidly and led to the development of several methods of treatment, including molecular targeted chemotherapy. Although several chemotherapeutic agents have proven to be quite effective in treating numerous patients with lung cancer, in the majority of cases, patients either already were resistant to the effects of the drug or eventually developed resistance as time progressed. Erlotinib, for example, typically is effective on patients with EGFR mutations, but generally does not successfully treat lung cancer patients without malignancy reoccurrence. Although numerous mechanisms for drug resistance have been developed, they are not able to effectively account for all cases of drug resistance. However, research has shown promising potential in targeting multiple tumorigenic pathways simultaneously. Combination therapy with erlotinib has already demonstrated success in numerous lung cancer cell lines. Antitumor effects have already been observed in several studies. Current studies are exploring new agents to enhance chemotherapy and focusing on identifying potential predictors of treatment benefit Leighl, . FurtherThe authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Patients who suffer from severe burns develop metabolic imbalances and systemic inflammatory response syndrome (SIRS) which can result in multiple organ failure and death. Research aimed at reducing the inflammatory process has yielded new insight into burn injury therapies. In this review, we discuss strategies used to curb inflammation in burn injuries and note that further studies with high quality evidence are necessary. Trauma resulting from severe burn injury triggers a systemic inflammatory response syndrome (SIRS) and serious metabolic disturbances. One of the best known systemic manifestations observed in the first hours after a major burn injuries is related to increased systemic capillary permeability with protein leakage into the interstitial space, generalized edema, and a tendency toward hypovolemic shock. Adequate fluid replacement is mandatory in the first hours after a traumatic burn. However, in burn patients, other systemic disorders are also accompanied by SIRS such as cardiac dysfunction, acute respiratory distress syndrome, acute renal failure, increased intestinal permeability resulting in bacterial translocation, hypermetabolism, hypercatabolism, and sepsis –4. TheseBurn injury induces global changes to the entire immune system resulting in suppressed immune function and increased susceptibility to infection. This immunopathological response can contribute to the development of SIRS and subsequent multiple organ failure. Patients with severe burns are more likely to die from sepsis due to the massive release of inflammatory mediators from the burn wounds. Total body surface area (BSA) involved and smoke inhalation are predictors of death. Each one percent increase in total body surface area burned was associated with a six percent increase in mortality risk. Also, the presence of smoke inhalation increased mortality risk by ninefold . In addiNevertheless, the systemic disorders observed in the first hours after a severe burn injury are related to increased systemic capillary permeability with protein leakage and a tendency toward hypovolemic shock. Burns greater than 10% BSA in children or 15% BSA in adults are potentially life-threatening injuries (because of the risk of hypovolemic shock) and should be treated with formal fluid resuscitation and monitoring in a burn unit . Hence, Despite the convenience of using formulas as an initial guide for fluid replacement ∗%  BSA), it is difficult to carry out fine adjustments in fluid delivery to the severely burned patient in practice. Commonly, there is a tendency to administer an oversupply of fluid (fluid creep) –9. But wIn the last decade, our burn center staff has preferred the use of the formula: 3 mL∗weight  (kg)∗% BSA of crystalloid infusion instead of the Parkland formula in the first 24 hours after burn injury. Using our formula, we have observed only minor amounts of general edema in the first days after extensive burn trauma, with consequent reduced morbidity and faster recovery. Our findings, “data not shown,” have been shared by other authors , 13. Fluα), interleukin-1b (IL-1b), interleukin-6 (IL-6), interleukin-8 (IL-8), and interleukin-10 (IL-10), have been reported both systemically and locally in burn patients. It has been proposed that there is a genetic influence on cytokine production and susceptibility to sepsis. A recent and interesting study indicates that genetically determined individual differences in IL-10 production might influence the susceptibility to septic complications in burned patients -1,3-linked glucose polymer derived from the cell wall of the yeast, revisiae , 29. Glua et al.  observedn injury . in vivo and in vitro and may serve as a therapeutic agent for the prevention of a systemic inflammatory response and leakage of fluid into tissue after major burn injuries [Ulinastatin is a protease inhibitor obtained from human urine; it has been shown to have anti-inflammatory effect by suppressing the production of proinflammatory cytokines . Luo et injuries . Coagulation and inflammation may interact during thermal injuries with major consequences for the pathogenesis of microvascular injury and subsequent multiple organ dysfunction or failure ; their iTreatment focused on the normalization of coagulation and the inhibition of systemic inflammation might have a positive impact on organ function and overall outcome in septic burn patients . MoleculTreatment of coagulation abnormalities is a challenge for burn specialists. Clear indications, efficacy, and the economic feasibility of the use of specific coagulation factors in burn patients should be targeted in clinical trials over the next few years.Nutritional support plays an undisputed part in the treatment of critically ill patients. Enteral nutrition supplemented with arginine, for example, alters intestinal homogenates from a pro- to an anti-inflammatory profile in mice with large areas of burn injury . NutritiStudies have shown that increased systemic retinol binding protein (RBP) levels are associated with insulin resistance (IR) and hyperinflammation in diabetic and obese patients. Increased RBP levels occurring after burn injury correlate with increased IR, inflammatory and catabolic responses, incidence of multiorgan failure, and mortality . InsulinSIRS is associated with a debilitating systemic hypercatabolic state, which is mediated by cytokines and chemokines . Systemiα) levels are lower in severe cases. These variations in cytokine levels may indicate impaired healing in severe burn injury patients which leads to their poorer prognosis. One protein that has been studied in inflammation control is leptin. Leptin is a circulating hormone that regulates energy intake and expenditure, including appetite and metabolism. It is well established that leptin is involved in the regulation of inflammation –51. LeptLeptin production is acutely increased during infection and inflammation, as a part of the host's acute phase response . On the Another hormone that should be mentioned in inflammation in burns is cortisol. Although hypercortisolemia has been suggested as a primary hormonal mediator of whole-body catabolism following severe burn injury, it may not play a central role in the postburn hypermetabolic catabolic response .Oxidative stress has been documented in burn injuries in both animals  and humaA patient's vitamin status is directly involved in inflammation, antioxidant response, burn wound healing, and the expected immune responses . AdjuvanIt is known that severe burns can be accompanied by vasoplegic syndrome (VS), a phenomenon which is characterized by persistent and diffuse vasodilation, hypotension, and low vascular resistance resulting in circulatory and respiratory failure. The decrease in systemic vascular resistance observed in VS is associated with excessive production of nitric oxide (NO). The plasma NO content is increased during the first hours after burn injury. It seems that the increased concentration of NO, combined with other biochemical phenomena of the systemic inflammatory response, leads to a widespread leakage of protein and intravascular fluid into the interstitial space, resulting in various degrees of edema and hypovolemia. Nitric oxide stimulates soluble guanylate cyclase to increase cyclic guanosine monophosphate (cGMP) production, leading to smooth muscle relaxation. The NO competitor, methylene blue (MB), which is an inhibitor of the soluble guanylate cyclase (sGC), has been proposed in the treatment of refractory cases of vasoplegia. We suggest MB as a viable, safe, and useful coadjuvant therapeutic tool to be used during fluid resuscitation , 68. Another way to suppress vascular hyperpermeability is the administration of nitric oxide synthase (NOS) inhibitors . HoweverThe intrinsic mechanisms by which inhalation injury contributes to elevated mortality are unclear. One hypothesis is that a massive influx of inflammatory mediators and cells  results in direct injury to the airway, leading to production of secretions and airway obstruction with mucus and products of inflammatory cell breakdown including extracellular deoxyribonucleic acid , 72.β1. Potential sources of extracellular DNA in airway secretions include directly damaged epithelial cells and inflammatory cells responding to injury or infection [Joyner et al.  found manfection . Therefonfection , 75. TheIn addition to the use of MSCs in regenerative research, it is known that these cells can also be used for modulation of inflammation. Bone-marrow-derived MSC allografts administered via intravenous transplantation have been shown to decrease proinflammatory cytokines, increase anti-inflammatory cytokines, decrease lung water mass fraction, ameliorate the systemic inflammatory response, and protect lung tissue in rabbits with smoke inhalation injury . MSCs maMSCs have a therapeutic benefit in burn-injured animals by providing anti-inflammatory as well as antiapoptotic effects. Human MSCs administered to the muscle of burned rats reduced infiltration of inflammatory cells into organs as kidney, lung, and liver. Interleukin-10 (IL-10) is one of the key cytokines with anti-inflammatory capacities; it has been demonstrated that MSCs can enhance the secretion of IL-10 by macrophages or dendritic cells. Furthermore, the activation of the AKT  signaling pathway in transplanted MSCs can provide an antiapoptotic effect in clinical settings of systemic inflammation .In an experimental study in rats, treatment with hyperbaric oxygen accelerated the recovery from burn wounds and reduced the development of scars . Rates oHypothermia has been implicated as an aggravating factor in critically ill patients, including those with serious burns , 85. On Peripherally active opioids have anti-inflammatory effects and can modulate wound healing. Local opioid application is being used for pain reduction in patients with inflammatory lesions such as burns . Opioid Burns are unique among acute injuries in the progressive nature of tissue necrosis and possible serious complications following the initial trauma, such as SIRS and severe metabolic imbalance. This intense instability in homeostasis may result in multiple organ failure and death. Current treatment of burn shock includes prompt fluid resuscitation and early burn wound excision. However, aggressive crystalloid-based resuscitation strategies are also associated with inflammatory mediator dysfunction. Hence, the patient's care team must always be careful to restrict the supply of liquid to only what is necessary to rescue the hypovolemic shock. Early excision of burns plays an important role in attenuation of SIRS as it can decrease the release of inflammatory mediators and the bacterial colonization of wounds. Over the past twenty years, since the introduction of early excision and grafting in our burn unit, we have observed a reduction in the mortality rate from 12% to less than 5%.Currently, alternative therapies are emerging that seek to modulate the complex systemic inflammation in burns. We support the discovery of new therapeutic options that can modulate the production of inflammatory cytokines which we believe will lead to improved treatment of burns. Inherently, the knowledge of the balance between pro- and anti-inflammatory cytokines is of fundamental importance and this also becomes a challenge to be overcome in the future. The aim of this paper is to comprehensively review current therapies available which can inhibit the inflammatory response to burns.