Patients with classic galactosemia, a genetic metabolic disorder, encounter cognitive impairments, including motor (speech), language, and memory deficits. We used functional magnetic resonance imaging to evaluate spontaneous functional connectivity during rest to investigate potential abnormalities in neural networks. We characterized networks using seed-based correlation analysis in 13 adolescent patients and 13 matched controls. Results point towards alterations in several networks, including well-known resting-state networks . Particularly, patients showed alterations in networks encompassing medial prefrontal cortex, parietal lobule and (pre)cuneus, involved in spatial orientation and attention. Furthermore, altered connectivity of networks including the insula and superior frontal gyrus -important for sensory-motor integration and motor (speech) planning- was demonstrated. Lastly, abnormalities were found in networks involving occipital regions, linked to visuospatial capacities and working memory. Importantly, across several seeds, altered functional connectivity to the superior frontal cortex, anterior insula, parietal lobule and the (pre)cuneus was observed in patients, suggesting special importance of these brain regions. Moreover, these alterations correlated with neurocognitive test results, supporting a relation with the clinical phenotype. Our findings contribute to improved characterization of brain impairments in classic galactosemia and provide directions for further investigations. Patients with classic galactosemia suffer from a profound deficiency of galactose-1-phosphate uridylyltransferase (GALT) due to mutations in the GALT gene, and affected newborns present with a severe toxicity syndrome upon exposure to galactose in milk. Immediate initiation of galactose-restriction is life-saving, but fails to prevent the development of burdensome brain impairments, including lower intelligence8, language production and speech (motor) problems13, slower information processing and memory deficits14, social difficulties15 and psychiatric conditions16. Furthermore, neurological sequelae are regularly reported, such as tremors, dystonia, coordination and balance disturbances, and ataxia18.The brain represents one of the target organs of damage in classic galactosemia, an inborn error of galactose metabolism, resulting in chronic impairments with significant impact on quality of life and general performance29. Using innovative diffusion-weighted magnetic resonance imaging (MRI) analysis techniques, evidence for abnormal white matter microstructure in bilateral anterior tracts (reduced neurite density) and the left hemisphere (increased dispersion of neurites) was provided28, which strengthens the hypothesis of aberrant myelin composition, possibly resulting from deficient galactocerebroside formation due to aberrant glycosylation26. Additionally, several studies demonstrated cerebral or cerebellar atrophy and decreased tissue density of grey matter29. This neuronal loss has been speculated to result from direct toxicity of galactose and its metabolites or aberrant glycosylation26. In addition to reduced grey matter density, Timmers et al. also observed increased grey matter density in the inferior frontal and medial prefrontal cortex, which could reflect compensation for problematic motor and cognitive functions  or abnormal maturation29.The pathogenic basis of these debilitations is puzzling and there are an increasing number of studies providing evidence for structural changes in white and grey matter, as well as alterations at a functional levelet al. used [18 F]fluorodeoxyglucose (FDG) positron emission tomography (PET) to evaluate glucose metabolism in patients21. Decreased metabolism was found in the occipital region, orbital frontal lobes, sensorimotor areas, cerebellum, superior temporal lobes, and the mid and superior parietal regions. Increased metabolism was demonstrated in bilateral anterior cingulate cortex (ACC) and temporal poles, as well as in basal ganglia. The findings in ACC and basal ganglia were interpreted as showing potential compensation for problematic motor functions.At a functional level, Dubroff 27. Evaluation of neural activity demonstrated that patients recruited different and more extensive brain regions during language production as compared to controls. The left inferior frontal gyrus (IFG), the right insula and the left pre-supplementary motor area (pre-SMA) were subsequently selected as seeds for a seed-based functional connectivity analysis, which revealed variations in functional connectivity in comparison to controls.Recently, a task-based functional MRI (fMRI) study by our group, wherein patients carried out a language production task, was the first to point towards abnormal language-related brain networks in this disorder33. A commonly used approach to study functional connectivity is the seed-based correlation analysis (SCA). This method is hypothesis-driven and correlates the resting-state time courses of an a priori selected region of interest  to the time series of the rest of the brain35, wherein the seed is selected from a task-dependent activation map or anatomically based on previous literature36. Despite differences in data acquisition and analysis methods, multiple resting-state networks have been consistently found across functional connectivity studies, such as the default mode network (DMN)38. Additionally, altered functioning of these networks has been demonstrated in participants suffering from a variety of brain disorders39, including neurodegenerative diseases such as Alzheimer’s disease42, as well as attention deficit hyperactivity disorder (ADHD)45 and other psychiatric conditions.Resting-state fMRI studies, in which neuronal connectivity in absence of a specific stimulus or task is assessed, are increasingly conducted within multiple fields of neuroscience to study the organization of core processing systems of the brain. The rationale behind these studies is that the brain is always active and shows spontaneous neuronal activity even during rest. Brain regions that show synchronous neuronal activity are considered to be functionally connected, thereby constituting a functional network27. However, resting-state fMRI allows us to investigate the overall functional organization of brain networks, which has not been studied before in classic galactosemia. The current study therefore characterized functional networks in resting-state in patients with classic galactosemia as compared to healthy controls using a seed-based approach. Seeds were selected based on previous neuroimaging studies in this population, namely the left IFG, right insula, left pre-SMA, right and left putamen, right and left medial prefrontal cortex (mPFC), and the medial occipital region29. Findings were correlated to available behavioral outcome measures as well, in order to confirm potential relations between functional networks and behavior46. For this, measures of functional connectivity of brain regions of interest were correlated to neurocognitive test results. Our aim was to elucidate affected brain networks in patients with this disease to improve the characterization of brain impairments encountered by these patients.In our previous study, we focused on functional networks that are involved in an active taskq(FDR) corrected map  and the bilateral posterior cingulate cortex (PCC) and precuneus (Pc) Fig. . Furthermap Fig. . More exmap Fig. , Table 1Connectivity maps showed that the left mPFC seed was functionally connected with the surrounding medial and superior frontal area, as well as with the dorsal ACC  in the control group Fig. . In patiThe seed in the right putamen showed temporal correlation with the head of the right nucleus caudatus and right thalamus, as well as with a small region of the right IFG Fig. . In the q(FDR) corrected map  corrected map revealed a less extensive network in patients, with less extensive correlation between the seed and its surrounding area, the contralateral side, and the left parietal lobule SMA and premotor cortices and the left PrG in the control group Fig. . A similActivity in the medial occipital region seed correlated with activity in bilateral middle occipital gyrus, bilateral lingual gyrus, extrastriate cortex, bilateral cuneus, the right precuneus, the right and left PrG in controls Fig. . In the r = 0.62, p = 0.03). This cluster showed increased functional connectivity in the patient group as compared to controls, which altogether suggests that increased connectivity correlates with lower performance (slower VOTs) in patients. Functional connectivity between the left mPFC and both right SPL and right precuneus were negatively correlated with the VOTs . These clusters showed decreased functional connectivity in patients as compared to controls, which altogether suggests that decreased connectivity correlates with lower performance (slower VOTs) on language production in the patient group. Lastly, functional connectivity between the right mPFC and right temporal lobe was positively correlated with VOTs . This cluster showed increased functional connectivity in the patient group as compared to controls, which altogether suggests that increased connectivity correlates with lower language performance (slower VOTs) in patients.A significant correlation between the patients’ beta values  and results of neurocognitive tests was found for eight clusters Table , Fig. 5Ar = −0.67, p = 0.02). This cluster showed decreased functional connectivity in patients as compared to controls, which altogether suggests that decreased connectivity correlates with better performance in patients. On the other hand, functional connectivity between the right mPFC and the left anterior insula was positively correlated with visuospatial working memory . This cluster showed increased functional connectivity in patients as compared to controls), which altogether suggests than an increased connectivity correlates with higher performance on visuospatial working memory in the patient group.Correlations between functional connectivity patterns and visuospatial working memory were observed as well, both involving the left anterior insula . Functional connectivity between the left mPFC and left anterior insula was negatively correlated with visuospatial working memory : decreased functional connectivity correlated with better performance in patients. As this cluster showed decreased connectivity in patients as compared to controls, this suggests that more dissimilar connectivity pattern to controls are associated with better performance in patients.Finally, verbal working memory was correlated to functional connectivity patterns too. Functional connectivity between the occipital region and left cuneus (cluster 1) was negatively correlated with verbal working memory , cuneus and anterior insula. These results indicate a special role of these brain regions within the altered brain connectivity in this patient group , and has been associated with introspective processes such as attention and consciousness, integration of cognitive and emotional information, and episodic memory56. The right insula seed analysis revealed this network in both groups, yet with altered connectivity in patients as compared to controls. Aberrant connectivity between the anterior insula and SFG/mPFC was observed across multiple seed analyses in this study. A meta-analysis of functional neuroimaging data indicates that the insula provides an interface between feelings, cognition and action, and can be subdivided in three parts with distinct functional connectivity profiles57: dorsal anterior insula , ventral anterior insula  and posterior insula . The altered connectivity between prefrontal regions related to executive functions and sensory-motor integration structures may relate to (compensation of) problems in the patients motor sequence planning, including motor speech production and planning difficulties60. This would be in agreement with previously described motor (speech) disorders in classic galactosemia61, as well as with the observed correlation between increased connectivity and poorer language production performance.The insula constitutes a key element of the salience network. This resting-state network additionally consists of the ACC as well as other subcortical and limbic structures, and is suggested to integrate external sensory information with internal emotional and bodily state signals in order to guide behavior, thereby engaging bottom-up detection, attention, working memory and motor systems62. We observed alterations in the occipital seed functional connectivity in the patient group. Evaluation of the occipital seed analysis showed decreased connectivity with the cuneus, which has been implicated in visuospatial attention/processing64. Findings from the right and left putamen analyses  are also in line with this hypothesis.The visual network, encompassing a primary visual component  and an extrastriate component , has been associated with processing of visual information66, as well as for social cognition68, showed abnormal connectivity in patients across several analyses. Previous studies from our group already demonstrated specific abnormalities in the language network leading to lexical and syntactic planning impairments, in addition to often observed motor speech planning problems27. The finding of an association between mPFC – temporal cortex connectivity and language production performance supports this further.Additionally, the STG and MTG, which are important for perception and production of language and speechThe correlations between the patients’ resting-state brain connectivity patterns and their performances on several neurocognitive tasks were used to further investigate the relation between brain networks and behavior. Note that we only selected those clusters that were different between patients and controls, and that correlations could only be explored for the patient group. Nevertheless, we observed several strong and specific correlations for several clusters, with correlations for two clusters surviving even a Bonferroni correction. The correlations should be interpreted with caution, but they might support the general functional interpretation of our resting-state analysis. The observed correlations seem to underline a brain network-behavior relation, although this needs to be confirmed in task-based fMRI studies targeting these specific neurocognitive functions. Most importantly, brain-behavior correlations were detected especially for those clusters that come back across multiple seed analyses as being divergently connected (compared to controls), namely the insula, parietal lobule and (pre)cuneus Fig. , therebyIn general, these findings shed new light on the brain impairments in classic galactosemia. Future studies are needed to investigate whether the alterations observed in this study are the result of targeted damage to these specific regions/networks, or whether these structures are equally affected as other regions of the brain but show a more prominent functional impairment. Alternatively, these differences in functional connectivity might reflect compensation mechanisms to cope with cognitive difficulties. In order to provide more detailed hypotheses on this, investigations of longitudinal data and a systematic comparison of results of structural and functional brain imaging studies are needed.27. The connectivity patterns and network alterations are in agreement with the current resting-state findings. The observed language network abnormalities are also in line with an earlier syntactic language planning study using event related potentials (ERPs), in which lexical and syntactic planning impairments were seen12. Results from an FDG-PET study pointed towards abnormalities of the superior temporal lobes, parietal regions, primary visual cortex, sensorimotor areas and frontal lobes21, brain regions that had a repeatedly altered functional connectivity in the current study. Imaging studies exploring brain structure in patients with classic galactosemia provided evidence for both white and grey matter abnormalities in line with the neurocognitive profile of patients29 and with the here reported results. Previous investigations in classic galactosemia pointed towards cerebellar involvement as well26. Although this brain structure was only partially included in the functional coverage of the fMRI scans, altered functional connectivity with the cerebellum was not observed in our study. This might be explained by the presence of only mild neurological signs and symptoms in our patient sample (as cerebellar dysfunction is associated with neurologic signs), though it should be noted that these were not systematically evaluated . In addition, the a priori selected seeds were all cortical, resulting in neural networks that are mainly (sub)cortical. Future studies specifically tapping into cerebellar connectivity are needed to shed more light on this topic.The current findings are in line with earlier studies in the field of classic galactosemia. A previous tasked-evoked fMRI study from our group already demonstrated altered functional connectivity in the language network, with specific alterations at the level of the IFG, insula and pre-SMA, suggesting impaired sensory-motor integration, motor (speech) planning deficits and suboptimal communication between frontal regions and temporal/parietal regions16, future studies exploring potential gender variations in functional connectivity are desired. Furthermore, the current study method is based on certain assumptions and focuses on a priori selected seeds, deriving from previous investigations. Future resting-state fMRI studies could consider an independent component analysis approach, which analyzes the overall pattern of functional connectivity without focusing on specific regions of interest71. In addition, we correlated functional connectivity to results on neurocognitive tests in order to evaluate potential brain-behavior relations. Since correlations could only be explored for the patient group, conclusions should be drawn with caution. These findings provide the first conjectures on potential brain-behavior relationships and the hypotheses generated by the current study require further testing in more dedicated study designs.One of the limitations of this study is its limited sample size, making this study of explorative nature. The small cohort size hampers solid conclusions on functional connectivity and confirmation of results in a larger sample is thus warranted. Moreover, gender was not equally balanced in our test sample  and this cohort is therefore not fully representative for the classic galactosemia population. Though cognitive performance does not seem to differ between male and female patientsTaken together, robust differences in functional resting-state networks between patients with classic galactosemia and age- and gender-matched controls were found. Results from our SCA point towards alterations in several networks, including some well-known resting-state networks . More specifically, abnormalities were revealed in networks encompassing the mPFC, parietal lobule and (pre)cuneus, which are involved in spatial orientation and attention. Furthermore, altered connectivity of networks including the insula and SFG, which are important for sensory-motor integration and motor (speech) planning/sequencing, was demonstrated. Lastly, abnormalities were found in resting-state networks involving the occipital region and cuneus, which might be linked to the observed impairments in visuospatial capacities and working memory. Most importantly, across several seeds, altered functional connectivity to the superior frontal cortex, anterior insula, parietal lobule and the (pre)cuneus was observed in patients, suggesting these brain regions might be of special importance and deserve more thorough investigations. In addition, these differences in network connectivity correlated with clinical test results in patients, supporting a relation between functional abnormalities and the clinical phenotype. Our findings contribute to the characterization of functional brain impairments in classic galactosemia, which is needed to create a better understanding of this enigmatic disease.et al.27. Briefly, patients who participated in an earlier study of our group and aged 14 years or older were invited to participate in the current study. Classic galactosemia diagnosis was confirmed by GALT enzyme activity measurement and/or GALT mutation analysis. One patient and one control subject were excluded because of extensive motion during scanning and a current health condition, respectively. Characteristics of included patients and controls are presented in Table Thirteen adolescent patients with classic galactosemia and thirteen age- and gender-matched healthy controls participated in this study. Participants were recruited as described by Timmers 27. During the resting-state scan (duration: 6 minutes) a fixation point was presented and participants were instructed to relax, not to think of something in particular and to keep their eyes open. Total duration of the experiment, including the acquisition of other data, was approximately 90 minutes. Afterwards, participants completed a short questionnaire addressing the difficulty of the session .To prevent excessive motion during scanning, participants practiced lying in a dummy scanner. After this practice session, participants received explicit instructions and were placed comfortably in the MRI scanner. Resting-state data were acquired, followed by a language task. The task-evoked functional connectivity pattern analyses were described elsewhereData were obtained on a 3 T Siemens MAGNETOM® Allegra head scanner using an 8-channel head coil, and a 3 T Siemens MAGNETOM® Trio whole body scanner using a 32-channel head coil . Data acquisition on two different scanners was necessary as a result of irresolvable technical issues with the Allegra scanner. Four patients and four controls were scanned on this MRI scanner. Parameters of the Allegra scanner and Trio scanner were identical unless otherwise specified.T1-weighted anatomical images were acquired using an ADNI MPRAGE sequence with 192 slices and 1mm iso-voxel resolution covering the whole brain . Functional T2*-weighted resting-state images were obtained using a standard echo-planar imaging (EPI) sequence covering the whole brain, except for cerebellum .p > 0.05 for all parameters), linear trend removal and spatial smoothing (Gaussian filter FWHM of 4 mm). Functional datasets were co-registered with anatomical data and normalized in Talairach space (3.5 mm iso-voxel). Anatomical data of all participants were averaged to create a group-based dataset.Data were analyzed using BrainVoyager QX version 2.8.4.2645 . For functional datasets, the first four volumes of each complete time series were discarded because of saturation effects. Pre-processing of the functional data included correction for slice time differences, 3D head motion correction , the extracted mean signal from the cerebral spinal fluid (CSF) and white matter (WM), and the global signal were Z-normalized and added as variables of no interest. Though the approach remains a subject of debate73, we decided to regress out the global signal since we are interested in localized network-specific neuronal activity, which could be obscured by unspecific global blood oxygenation level dependent (BOLD) fluctuations. Furthermore, signal oscillations at a frequency of 0.1–0.25 Hz (sine-cosine pairs) were added as confounders for low-pass filtering of the time series.We conducted a SCA with the left IFG, right insula, left pre-SMA, right and left putamen, right and left mPFC, and the medial occipital region as seeds  < 0.05. For contrasts across groups, an initial threshold of p < 0.01 was chosen after which a cluster-size thresholding was performed using MonteCarlo simulations (n = 1000) to correct maps for multiple testing at the level of alpha 0.05.The BOLD time courses of the seeds were extracted, normalized and correlated with the time series from all other brain voxels using a random-effects (RFX) group analysis. We first estimated functional connectivity for each participant separately in a multi-study RFX General Linear Model analysis (first level), which then served as input for a second-level analysis of variance (ANOVA), allowing group contrasts. Functional connectivity per group was inspected at the level of the whole cerebrum on maps corrected at 12) using a Pearson correlation analysis. The Immediate Recall subtest of the Rey Osterreith Complex Figure (expressed as T-score) was used to evaluate visual working memory74. The Digit Span (Forward and Backward) measured verbal working memory75. The Bourdon-Vos test was used to assess sustained attention (mean reaction time [RT])76. Voice onset times (VOTs) from a sentence production task of a previous study in the same patient group were used as a measure of language production12. Correlations with a p value < 0.05 were considered statistically significant.From clusters in which group differences were observed, beta values were extracted. In the patient group, the beta values per cluster  were correlated with results of neurocognitive tests (performed prior to the current study as part of a different investigation by our group


Abra segmentum through multiple sea-level fluctuations preserved in brackish Holocene deposits of the Po Plain, Italy. Prevalence values were significantly elevated (p < 0.01) in samples associated with flooding surfaces, yet the temporal trends of parasite prevalence and host shell length, cannot be explained by Waltherian facies change, host availability, salinity, diversity, turnover, or community structure. The observed surges in parasite prevalence during past flooding events indicate that the ongoing global warming and sea-level rise will lead to significant intensification of trematode parasitism, suppressed fecundity of common benthic organisms, and negative impacts on marine ecosystems, ecosystem services, and, eventually, to human well-being.The forecasts of increasing global temperature and sea level rise have led to concern about the response of parasites to anthropogenic climate change. Whereas ecological studies of parasite response to environmental shifts are necessarily limited to short time scales, the fossil record can potentially provide a quantitative archive of long-term ecological responses to past climate transitions. Here, we document multi-centennial scale changes in prevalence of trematodes infesting the bivalve host  However, ecological research on the response of parasites to anthropogenic warming is necessarily limited to short time scales of the most recent months and years8. In this respect, the most promising avenue is offered by the latest Quaternary fossil record where, thanks to highly-resolved sequence and chrono-stratigraphic frameworks16, past parasite-host interactions can be examined over geologically short (102–103 years), societally relevant time scales.Understanding the historical impact of climate variability on heterocious parasites is prerequisite for forecasting parasite-host interactions in the near future and assessing the potential implications for ecosystem health, ecosystem services, and human well-being15. It is usually in the second intermediate host stage that digeneans affect shell secretion in their molluscan hosts. Gymnophallids induce the active growth of characteristic pits with raised rims on the interior of their bivalve host’s shells , is always a mollusk species. The newly emerged cercariae larvae infest the second intermediate host where the parasite is in an encysted, latent stage (metacercaria), waiting to be ingested by the third, definitive host, which is always a vertebrate organism that enables sexual reproduction of the adult parasite. A peculiarity of many trematode species, within the family Gymnophallidae, is that they have the same first and second intermediate individual hosts, skipping the intermediary, free-living cercariae stage23 demonstrated that trematode prevalence peaked in the lower part of paralic transgressive deposits recording the generalized inundation of the regional coastal system coincident with Meltwater Pulse 1c, that took place between 9.5 and 9.2 ky24. Similarly, significantly higher trematode prevalence was documented in host taxa from sediment-starved northern Adriatic strandline death assemblages, relative to that documented from comparable assemblages from the Po delta shoreline25. These two coastal regimes serve respectively as modern analogues for Holocene transgressive and prograding settings26. This putative link between overall sea-level rise and prevalence, if demonstrated on societally relevant time scales, could serve as an analogue for the response of parasitism to global warming in the coming decades to centuries. Here we explicitly test the link between short term (102–103 years) flooding pulses and upsurges in parasite prevalence using the fossil record of bivalve hosts from a cored Holocene back-barrier succession  time scales (Online Methods). The strata in core 204-S7, from bottom to top, are composed of stacked fluvial channel facies associations  transgressive-regressive sequence of the coastal Po Plain is a wedge-shaped genetically related package of latest Pleistocene-to-Holocene strata. In the studied core 204-S7, the T-R sequence is a few tens of meters thick Fig. , with it30.We collected 61 bulk samples from the top 20 meters of core 204-S7 Fig. . A non-m27 representing short-lived, rapid transition from mesohaline to polyhaline dominated environments . Anterior-posterior length of A. segmentum ranged between 1.5 and 22.3 mm  abundance and prevalence , likely due to the accumulation of parasites through ontogeny. Prevalence values were positively and significantly correlated with host shell length , however there were no significant associations between shell length and either flooding pulses or nMDS1 scores . All Abra valves were classified as either whole or broken, and the broken valves were further categorized into either “sufficiently complete” or “too fragmented” to determine infestation status. There was no significant difference in trematode prevalence values of whole and “sufficiently complete” broken valves . These results suggest that the proportion of “too fragmented” valves was unlikely to represent an important confounding factor in reconstructing the stratigraphic record of trematode dynamics.Sample nMDS1 scores were negatively correlated with the proportion of 40. Therefore, we hypothesize that gymnophallid trematode prevalence will be more strongly influenced by the creation of new habitat in brackish and freshwater settings than in shallow marine settings. Though not the direct topic of research here, an increase in wetlands created by sea level rise would generate new habitat for the gastropod intermediate hosts of Schistosoma41, the trematodes responsible for schistosomiasis in humans.Another potential factor affecting parasite prevalence is the fluctuating availability of habitat-area for trematodes during sea-level cycles. The geologically rapid creation of new habitat during flooding pulses and their subsequent destruction during progradation could exert a first order control on trematode prevalence during high frequency cycles. As sea level continues to rise, some settings will be more strongly influenced than others. For instance, densely populated lowlands, estuarine, and riverine settings would likely display the greatest increase in trematode habitat-area during relative sea level rise as a direct effect of flooding and, indirectly, by the landward rise of the groundwater tableThe fossil record of the northern Adriatic points to a significant association between the prevalence of heterocious parasites and flooding events recording repeated climate-driven sea level shifts. From this historical perspective we posit that the ongoing anthropogenic warming and sea-level rise should trigger a significant upsurge in gymnophallid trematode prevalence and the expansion of wetland habitats ideal for schistosomatid intermediate hosts. The forecasted changes are expected to suppress the fecundity of common benthic organisms, exert negative impacts on ecosystems, impede ecosystem services, and, eventually, negatively affect human well-being.Online Supplementary Info


Geronticus eremita), a critically endangered bird. In total, 43 individually marked focal animals from a free-ranging colony were investigated. The analyses included a description of initiated and received affiliative interactions, pair bond status as well as seasonal patterns of hormone and endoparasite levels. During the reproductive season, droppings contained parasite eggs more often and corticosterone metabolite levels were higher as compared to the period after reproduction. The excretion rate of endoparasite products was lower in paired individuals than in unpaired ones, but paired animals exhibited higher corticosterone metabolite concentrations than unpaired individuals. Furthermore, paired individuals initiated affiliative behaviour more frequently than unpaired ones. This suggests that the reproductive season influences the excretion patterns of endoparasite products and corticosterone metabolites and that affiliative interactions between pair partners may positively affect endoparasite burden during periods of elevated glucocorticoid levels. Being embedded in a pair bond may have a positive impact on individual immune system and parasite resistance.The reproductive season is energetically costly as revealed by elevated glucocorticoid concentrations, constrained immune functions and an increased risk of infections. Social allies and affiliative interactions may buffer physiological stress responses and thereby alleviate associated effects. In the present study, we investigated the seasonal differences of immune reactive corticosterone metabolite concentrations, endoparasite burden (nematode eggs and coccidian oocysts) and affiliative interactions in northern bald ibis ( Rupicapra rupicapra) showed elevated cortisol metabolite concentrations and parasite levels during the mating season as compared to the post-mating period [Peromyscus maniculatus and P. leucopus; [Petrochelidon pyrrhonota; [Anser anser) parasite excretion decreased throughout the parental season, which has been suggested to be related to the high energetic costs of the reproductive season [Parus caeruleus) increased their feeding rates in compensation [Sturnus unicolor) with parasitized nests [The reproductive season is considered socially and energetically costly when compared to the post-reproductive one ,2. Such g period . Chronicg period  and the g period –10. The eucopus; ) and clirhonota; ) showed e season . Furthere season ,15. Whenensation , whereased nests ,16.Macaca sylvanus; [Pan troglodytes schweinfurthii, [, the presence of the mother buffered the stress response to an aversive stimulus [Geronticus eremita) showed that paired individuals excreting high levels of endoparasite products during the reproductive season also engaged in more allopreening behaviour [Social allies and affiliative behaviour may help buffering physiological stress responses –21. For ylvanus; ) and wilfurthii, ) showed stimulus  and in gstimulus ,18,19. Astimulus , indicatstimulus . A recenehaviour .We presently aim at investigating the seasonal differences of concentrations of excreted immune reactive corticosterone metabolites (CM), parasite burden (as measured by the number of samples containing endoparasite products) and frequencies of affiliative behaviour between the reproductive and post-reproductive season in the free-roaming and individually marked colony of northern bald ibis at the Konrad Lorenz Research Station (Austria). This seasonally monogamous and year-round colonial species breeds in dense colonies and forages in flocks, that often split into subgroups of different sizes . Pair boWe distinguished between initiated and received affiliative behaviour since they may differ in their effects on glucocorticoid secretion, as measured by a reduced excretion of CM , self-diThis study complies with all current Austrian laws and regulations concerning the work with wildlife. Catching, restraining of the birds, observing the animals and collecting droppings were performed under Animal Experiment Licence Number 66.006/0026-WF/V/3b/2014 by the Austrian Federal Ministry for Science and Research . We confirm that the owner of the land, the Duke of Cumberland, gave permission to conduct the study on this site. All data were collected non-invasively. Birds were habituated to the presence of humans.males = 24, Nfemales = 19). The age ranged from 0 to 16 years (mean: 4.2). According to the classification described by Böhm and Pegoraro [The study was conducted in Grünau im Almtal . The free-ranging colony of northern bald ibis was established in 1997 at the Konrad Lorenz Research Station (KLF) in Grünau im Almtal by hand-raising zoo-bred chicks  in coordPegoraro  focal inBehavioural data and individual droppings were collected over a period of 112 days between 15 May 2015 and 31 October 2015. Days of data collection were irregularly distributed over the sampling period, as the northern bald ibis population flew to Molln  and stayed there from August until October. In Molln, standardised data collection was not possible, due to inaccessible foraging grounds (e.g. cow paddocks). Hence, data collected in Molln were excluded from the analyses. We included both the reproductive and post-reproductive season in this study. The reproductive season was determined as the time from rearing to fledging of the offspring , whereas the post-reproductive season lasted from mid-June to October. Due to individual variations in the start of egg-laying, beginning and end of both periods were determined separately for each breeding pair depending on the hatching date of the offspring.www.cybertracker.org, [The behaviour was measured by applying a continuous recording method ,38 for eker.org, ). The frker.org, ). In sumDroppings of the focal individuals were collected in order to determine the amount of (1) CM and (2) excreted endoparasite products (nematode eggs and coccidian oocysts).Eudocimus albus) which are similar in size and diet [Steroid metabolites in droppings represent an integrated, proportional record of the unbound portion of plasma steroids depending on the frequency of defecation and gut passage time . We assuand diet . The same below)  at the lBesides the data collection described above, we compared several available enzyme immunoassays in order to select the best-suited one for measuring corticosterone secretion in the northern bald ibis, a necessary step when using such non-invasive methods for measuring faecal glucocorticoid metabolites ,45.To evaluate which of the currently available EIAs is best suited for measuring adrenocortical activity of the northern bald ibis, a handling experiment was run on two experimental days in December 2015 and January 2016. Four individuals  was done using a flotation method in a McMaster counting chamber . At leasDue to generally low parasite egg and oocyst abundance in the samples , we treated samples either as containing or not containing nematode eggs or coccidian oocysts.All statistical analyses were conducted using R version 3.2.5  and the To assess which factors influence (1) parasite burden (samples containing nematode eggs and coccidian oocysts–yes/no), (2) initiated (yes/no) and (3) received (yes/no) affiliative behaviour  we used GLMMs with binomial error distribution and logit link function. To analyse which factors influence (4) CM (response variable) we used a GLMM with gamma error distribution and inverse link function. Fixed factors in these models were (1) sex, (2) age, (3) pair bond status  and (4) season (reproductive and post-reproductive season). Since the northern bald ibis is a social, seasonally monogamous and colonial species, affiliative interactions can be observed between all colony members. Therefore, we also included pair bond status as a fixed factor in the affiliative behaviour models. Subject identities were included as random factors in all GLMMs to control for between-subject variation and unbalanced design. Field constraints did not allow a regular data collection for the three data sets . Interactions between fixed factors were not considered in the statistical analyses as they did not improve the model fit.To select the best models, we used an information theoretic approach and calculated all possible candidate models . We rankModel-averaged results identified season as the strongest determinant of the excretion patterns of nematode eggs and coccidian oocysts. The number of droppings containing endoparasite products was highest during the reproductive season as compared to the time after reproduction . While sPair bond status was the second-most important variable influencing the excretion of coccidian oocysts. Furthermore, pair bond status was indicated as the third-most important variable for nematode egg excretion, CM levels and initiated affiliative behaviour. Paired individuals excreted fewer droppings containing endoparasites  and exhiWe found that initiated affiliative behaviour and the excretion pattern of nematode eggs were best explained by sex. Males initiated affiliative interactions more frequently and excreted a higher number of droppings containing endoparasite products than females. The relative importance indicated the variable sex to be less important for coccidian oocysts. Furthermore, sex had no influence on CM.The most influential variable on the excretion patterns of CM was age, with juveniles exhibiting higher CM levels compared to adults, whereas sub-adult individuals showed lower concentrations than adults. Furthermore, it was the second-most influential variable for initiated affiliative behaviour. A higher frequency of initiating affiliative behaviour was found in juveniles compared to adults, whereas sub-adults initiated less than adults. Age had the least importance when considering the excretion patterns of coccidian oocysts, with lower excretion rates in juvenile and sub-adult individuals as compared to adult ones. Age, however, did not predict the excretion rate of nematode eggs.All candidate models with received affiliative behaviour as response variable did not improve penalised model fit over the null model, as assessed by AICc, indicating that variation in the data cannot be explained by any of these factors .Statistical details of the top-ranked models are presented in Tables We found different modulation patterns of excreted immune reactive corticosterone metabolites (CM) and endoparasites in the reproductive season as compared to the post-reproductive period in the seasonally monogamous but year-round social northern bald ibis. The reproductive season is energetically costly in terms of egg-laying, incubation and rearing of the offspring , which wAs our data are correlative, we can only speculate about a potential causality between elevated CM concentrations and endoparasite burden. We suggest that the increased excretion of endoparasite products as well as high corticosterone metabolite concentrations during the reproductive season may be the result of a trade-off between reproductive effort and immune function . High CMWe further found that males produced more endoparasite-positive droppings than females, suggesting that particularly the male immune system was affected by social investment. This is also indicated by the fact that males in general, independently of age, initiated more affiliative interactions compared to females, which would be expected to function as social support buffering females glucocorticoid secretion. As males are the donors of social support, which benefits the females, they also may generally be more susceptible to infections with parasites due to reduced immune functions as compared to females ,63. AnotAphelocoma coerulescens), while the oldest individuals exhibited again greater corticosterone levels [Higher CM levels and excretions of endoparasite products were found in adults compared to sub-adult individuals and adults also initiated more affiliative interactions. This may be linked to sexual maturity and hence to the necessary behavioural and physiological investment into reproduction. As glucocorticoids are the major hormones regulating metabolism , this ine levels .Probably due to relaxed pair bond relationships outside the reproductive season in the northern bald ibis , affiliaTaeniopygia guttata) for example, elevated corticosterone concentrations returned to baseline levels after reunion with the pair partner [Rattus norvegicus) did not show fear-related behaviours, such as freezing, when smelling the odour of a familiar conspecific [Saimiri sciureus) females showed decreased basal cortisol levels in the presence of a female pair partner [Contrary to expectations, we found no link between receiving affiliative behaviour and the excretion of CM and endoparasite products. However, the excretion patterns of endoparasite products were still lower in paired than unpaired individuals. This may indicate that initiating affiliative interactions with social partners may up-regulate immune functions, even though high CM levels are still present. In fact, pair bonded northern bald ibis excreted higher concentrations of CM compared to unpaired ones. Moreover, long-term monogamous avian species, such as greylag geese, show decreased excretion of CM when family members are present during stressful situations ,72. In s partner . This im partner ,74,75. S partner .On the other hand, it might be expected that interacting with parasitized conspecifics increase the risk of infection . HoweverA previous study on northern bald ibis showed that more nematode eggs were excreted in females than in males during the reproductive season , whereasWe conclude that excretion patterns of endoparasite products and CM concentrations differ in the colonial and seasonally monogamous northern bald ibis according to season. Even though reproduction is energetically costly and may be accompanied by elevated glucocorticoid concentrations ,2, affilS1 Dataset(CSV)Click here for additional data file.S2 Dataset(CSV)Click here for additional data file.S3 Dataset(CSV)Click here for additional data file.S1 TableName, sex , year of hatching, age class, pair bond status as well as involvement in the assay comparison are indicated.(DOCX)Click here for additional data file.


Saccharomyces cerevisiae proliferates in a logarithmicfashion when growing in glucose-containing rich media. When all carbon sources in theenvironment are depleted, the yeast cells enter the stationary phase. There are twotypes of stationary cells: the quiescent (Q) cells, which temporarily cease dividingbefore nutrition replenishes, and the non- quiescent (NQ) cells that keep proliferating2The budding yeast et al. reports that distinct granules are formed in Q and NQ cells,which determines their respective cell fates etal. also proved that the Q cells reenter the mitotic cycle more quicklyafter nutrition replenishment Cells within the stationary phase are commonly observed to have granule structures,containing stress response-related factors A key question remains to be addressed in the field: what determines a cell to becomequiescent or non-quiescent ? The molecular pathways that regulate this cell-fatecommitment remain elusive. Davidson et al.


It is undeniably one of the greatest findings in biology that (with some very minor exceptions) every cell in the body possesses the whole genetic information needed to generate a complete individual. Today, this concept has been so thoroughly assimilated that we struggle to still see how surprising this finding actually was: all cellular phenotypes naturally occurring in one person are generated from genetic uniformity, and thus are per definition epigenetic. Transcriptional mechanisms are clearly critical for developing and protecting cell identities, because a mis-expression of few or even single genes can efficiently induce inappropriate cellular programmes. However, how transcriptional activities are molecularly controlled and which of the many known epigenomic features have causal roles remains unclear. Today, clarification of this issue is more pressing than ever because profiling efforts and epigenome-wide association studies (EWAS) continuously provide comprehensive datasets depicting epigenomic differences between tissues and disease states. In this commentary, we propagate the idea of a widespread follow-up use of epigenome editing technology in EWAS studies. This would enable them to address the questions of which features, where in the genome, and which circumstances are essential to shape development and trigger disease states. In hual cells ). Despitepigenetic phenotypes, although studied for decades now, is still avidly debated. In the centre of attention lie epigenomic processes, reversible marks, modifications and features of chromatin, a macromolecular structure consisting of unmodified and chemically modified nucleic and amino acids found in the nucleus. Chromatin consists first and foremost of DNA itself, whose bases—primarily cytosine residues—can be chemically modified adding information to the DNA sequence [Which processes are causally inducing or protecting sequence . Chromatepigenomic marks possess all the prerequisites to control epigenetic phenotypes, but probably only a small fraction of those found in the nucleus might indeed possess this regulative power.However, with respect to their contribution to gene silencing, we face a paradoxical situation. On the one hand, these marks correlate with inactive loci, the enzymes setting these marks are usually necessary for normal animal development, and well-studied model systems exist proving their functionality in some cases : genomic2.Over the last decades, a plethora of methods has been developed to facilitate epigenomic profiling. In principle, these approaches can be subdivided into two categories by the genomic resolution they offer. Single-base resolution approaches are commonly used to map and quantify DNA modifications. Cytosine methylation (mC) can be detected by bisulfite conversion resulting in the chemical deamination of unmethylated cytosine to uracil while methylated cytosine is protected. After PCR amplification, DNA methylation can be mapped and quantified at single-base resolution by massive parallel sequencing or array hybridization . Profilihttp://www.roadmapepigenomics.org/protocols).While being rather cost-intensive in the beginning, recent technological advances in array technology and sequencing have made large epigenome profiling projects feasible. Large consortia such as ENCODE (Encyclopedia of DNA Elements), REMC (Roadmap Epigenomics Mapping Consortium) and IHEC  are generating comprehensive epigenomic datasets establishing important references relevant to human health and disease –30. Span3.Sequencing of the human genome 15 years ago triggered a global effort to uncover genetic causes of human disease. Since then, many genome-wide association studies (GWASs) have been conducted to identify disease-causing variants hidden in the human population. Although these efforts have been highly successful in detecting disease-associated variants, these genetic aberrations often lacked meaningful predictive value ,32. Whila). Most of these studies have concentrated on DNA methylation largely due to the low amount of tissue required and the cost-effectiveness of detection methods such as Illumina's 450 K array [Technological advances have facilitated the genome-wide examination of epigenetic modifications in the context of disease phenotypes. Akin to GWASs, these epigenome-wide association studies (EWASs) make use of comprehensive cohorts of patients and control groups for large-scale association analysis of epigenomic marks in the context of disease phenotypes. EWAS publications have soared in recent years, primarily focusing on multifactorial disorders including cancer, neurodegenerative and autoimmune diseases a. Most o K array . More re K array , which a K array , systemi K array ,39. With K array , a thoroin vivo and in vitro models of genetic variants, techniques for testing the relevance of individual epigenomic marks have been completely lacking. However, Cas9-based approaches developed in recent years now offer the unique chance to functionally test EWAS hits and to start dissecting cause and effect of epigenomic changes associated with human disease.Taken together, EWASs have yielded thousands of epigenomic markers associated with human diseases, but causes of the disease-associated epigenomic changes remain largely unclear, a challenge that has long been also recognized for GWASs . Whereas4.Escherichia coli, approximately half of all bacteria (approx. 40%) and nearly all archaea (approx. 90%) are equipped with a sophisticated adaptive defence mechanism called clustered regularly interspaced short palindromic repeats (CRISPR) or CRISPR-associated (Cas) [cas genes, organized in operons, and a CRISPR-locus harbouring an array of genome-targeting sequences (termed spacers), which are derived from invading foreign DNA and flanked by identical direct repeats. CRISPR/Cas systems are classified into four different types with class II being the best studied and most widely adopted system as a molecular tool [trans-activating RNA (tracrRNA). By fusing the crRNA and tracrRNA into a single-guide RNA (sgRNA) [First observed in ed (Cas) . These slar tool . In type (sgRNA) , the RNA (sgRNA) –49. Howeb). Once the modifier is bound to the target site immediate consequences of the change in epigenomic marks can be monitored. Direct fusions of dCas9 with effectors have already been successfully employed to shed light on the causal relationship between epigenomic marks and levels of transcription. For instance, recruitment of the catalytic core of the human acetyltransferase p300 to promoters or enhancers leads to robust transcriptional activation [The principle is as simple as it is elegant. A chromatin modifier is fused to dCas9 and recruited to a chosen genomic site by a specific sgRNA b. Once ttivation . Additiotivation ,51. By ctivation –55. Thestivation . This aptivation ,58. An etivation ,60. Tran5.Despite the growing amount of profiles and EWAS data available, the functional value of associated hits remains elusive. At least for DNA methylation, a large number of high-quality profiles with disease-associated changes and a CRISPR toolbox to manipulate those have been reported. We propose to enter the post-epigenomic stage by choosing a number of candidate features from the high-quality epigenomic data generated to date and test their relevance by epigenome editing . For somin vitro. Essential for this is an experimental system that allows manipulation of the epigenomic mark and modelling of the human disease or phenotype  or together in a follow-up epigenomic screen using specially designed libraries of gRNAs. Although a perfect model system might rarely exist—diseases are transformations of organs not cells—certain aspects can often be reproduced in vitro. Moreover, the first attempts to transfer epigenome editing into the living animal have already been reported [in vitro, for example, if their effects depend on a very specific genetic background. We think, however, likely incidences of false negative results for some should not discourage from producing causal evidence for many other EWAS hits or profiling peaks.Several reviews have already suggested the use of epigenome engineering to validate or screen EWAS hits or profile peaks ,61. Testhenotype . Manipulreported ,66. It iAlthough the suggested procedure has not yet been adopted and the functional relevance of thousands of published EWAS hits remains unclear, several publications support the feasibility of editing DNA methylation as well as the functionality for some tested marks ,68. More


Subjective cognitive impairment can be a significant and prevalent problem for gynaecological cancer survivors. The aims of this study were to assess subjective cognitive functioning in gynaecological cancer survivors after primary cancer treatment, and to investigate the impact of cancer treatment on brain structural networks and its association with subjective cognitive impairment.This was a cross-sectional survey using a self-reported questionnaire by the Functional Assessment of Cancer Therapy-Cognitive Function (FACT-Cog) to assess subjective cognitive functioning, and applying DTI (diffusion tensor imaging) and graph theoretical analyses to investigate brain structural networks after primary cancer treatment.p values <0.001). Compared with patients who had received surgery only and non-cancer controls, patients treated with chemotherapy indicated the most altered global brain structural networks, especially in one of properties of small-worldness (p = 0.004). Reduced small-worldness was significantly associated with a lower FACT-Cog total score . Increased characteristic path length was also significantly associated with more subjective cognitive impairment .A total of 158 patients with gynaecological cancer  and 130 age-matched non-cancer controls  were assessed. Patients reported significantly greater subjective cognitive functioning on the FACT-Cog total score and two subscales of perceived cognitive impairment and perceived cognitive ability  in gynaecological cancer patients. As primary cancer treatment can result in a more random organisation of structural brain networks, this may reduce brain functional specificity and segregation, and have implications for cognitive impairment. Future prospective and longitudinal studies are needed to build upon the study findings in order to assess potentially relevant clinical and psychosocial variables and brain network measures, so as to more accurately understand the specific risk factors related to subjective cognitive impairment in the gynaecological cancer population. Such knowledge could inform the development of appropriate treatment and rehabilitation efforts to ameliorate cognitive impairment in gynaecological cancer survivors. Cognitive impairment can be a significant and prevalent problem for survivors with gynaecological cancer , 2. CognCRCI may be related to a number of psychological factors that are seldom investigated in the context of gynaecological cancer, although gynaecological cancer is the second most prevalent form of cancer among women in China . PsycholSelf-reported cognitive impairment are also associated with structural and functional changes in the brain that can be detected by magnetic resonance imaging (MRI) , 17. BraIn view of the poor understanding of the psychosocial impacts of cancer treatment on perceived cognitive impairment and brain networks in gynaecological cancer survivors, it is important to explore CRCI and its associated factors in this population. Therefore, this study aims to assess subjective cognitive functioning in gynaecological cancer survivors after primary cancer treatment, and to investigate the impact of cancer treatment on brain structural networks and cancer treatment’s association with cognitive impairment. This study also aims to explore associated predictors of subjective cognitive impairment in order to gain a deeper understanding of potential factors of importance to CRCI.This cross-sectional study was conducted to assess research participants’ subjective cognitive functioning, psychological wellbeing and brain structural networks immediately after primary cancer treatment. Age-matched non-cancer controls were simultaneously assessed using these outcome measures for direct comparison.Patients were recruited in South China, at a tumor hospital and at a general hospital’s gynaecological oncology unit. This study obtained ethical approval from the ethics committees at both Hunan Cancer Hospital and The Third Affiliated Hospital of Guangzhou Medical University. Subjects were Chinese females ages 18 to 60 years; with a primary diagnosis of gynaecological cancer, without metastatic disease. Patient exclusion criteria were women who did not have a primary diagnosis of cancer, and/or who were in a terminal stage of cancer. Advertisements to recruit non-cancer controls were posted in the hospitals’ common areas. Each non-cancer control subject was within 2 years of the age of the patient. All participants had to be without a diagnosis of a neurodegenerative disease or any potential psychiatric disorder, and without the use of psychotropic medication. All study participants provided written informed consent to participate.Subjective cognitive functioning was assessed using the Functional Assessment of Cancer Therapy-Cognitive (FACT-Cog) scale. A self-report questionnaire measures perceived cognitive impairment, comments from others, perceived cognitive ability, and impact of cognitive impairment on quality of life . The FACDepression and anxiety were evaluated using the Chinese version of the Hospital Anxiety and Depression Scale (HADS) . The HAD2and a b-value of 0. T1-weighted imaging was achieved for morphometric  analysis using three-dimensional fast spoiled-gradient recalled acquisition in steady state in 166 coronal slices .The MRI data were acquired using a Philips 3 T Achieva MRI scanner with an 8-channel head coil. Structural brain networks were assessed in 30 participants who reported cognitive complaints (with the FACT-Cog summary score ≤ 85): ten out of 130 healthy controls had a FACT-Cog summary score of less than 85, then 10 age-matched surgery patients and 10 age-matched patients with chemotherapy who also reported cognitive complaints were included in brain MRI scanning.. This cut-off score for the FACT-Cog was based on existing published studies , 29. DTIp value of 0.1 in the univariate analysis). Analysis of variance (ANOVA) was used to compare three participant groups reported with subjective cognitive impairment and brain structural network properties. Associations between brain network properties and subjective cognitive impairment were explored using Pearson’s correlation coefficient. The threshold of P < 0.05 was used to assess statistical significance.Data related to self-reported cognitive and psychological measures were analysed using SPSS for Windows . Descriptive, comparison and regression analysis were used to analyse behavioural data. Descriptive statistics were used to describe sociodemographic and clinical characteristics of the sample. In comparing subject characteristics, cognitive and psychological measures between patients and healthy controls used independent t-tests for continuous variables, and chi-square or Fischer exact tests were used for testing differences in categorical measures. To analyse the relationship between perceived cognitive functioning and associated factors, a univariate analysis was used followed by multiple regression analysis. As sociodemographic factors, such as age and education level, have been found to be associated with cognitive functioning in ovarian cancer survivors , it is ihttps://www.nitrc.org/projects/panda/). Each individual’s DTI dataset was registered to the same individual’s high-resolution structural image and then into the standard Montreal Neurological Institute (MNI) space using affine transformations. Fractional Anisotropy (FA) images were created from the pre-processed DTI data of all subjects. All FA images were then non-linearly aligned to a common space. The mean FA image was used to represent the centre of all tracts common to the group. Then, all subjects’ aligned FA data were projected onto the skeleton, and the resulting data were subjected to voxelwise cross-subject statistics. Whole brain tractography was then performed in the patient’s native space for each subject at each time point using a deterministic streamlined approach [The DTI images were preprocessed using PANDA: a pipeline toolbox for analysing brain diffusion images . The voxelwise brain structural network analyses were performed using the CPU-GUI platform of GRETNA [The assessment of brain network measures was performed using the toolkit of graph theoretical network analysis (GRETNA)  were in the early stages of cancer, more than 60% of patients  had a diagnosis of cervical cancer, and more than half of patients were receiving chemotherapy or a combination of chemotherapy and other cancer treatment. All research subjects’ demographic and clinical characteristics are shown in Table Of the 288 participants, 158 patients with gynaecological cancer had completed primary cancer treatment within a week, and 130 noncancer controls were balanced in terms of age and marital status (Table p < 0.001), especially in the subscale scores of perceived cognitive impairment and perceived cognitive ability . Within the patient group, patients receiving chemotherapy scored lower in the FACT-Cog total scores and four subscale scores  .While there were no statistically significant differences between patients and healthy controls in terms of anxiety, depression, and fatigue levels Table , there wy) Table . Associap = 0.004). Patients receiving chemotherapy had the lowest mean small-worldness index, compared with patients who received surgery only and healthy controls. Lower small-worldness index was associated with more subjective cognitive impairment  . Shorter characteristic path length, which indicates more efficient network organisation, was significantly associated with fewer subjective cognitive impairment  (Fig. From Table This was a cross-sectional survey using a self-reported questionnaire to assess subjective cognitive functioning, and applying DTI and graph theoretical analyses to investigate brain structural networks after primary cancer treatment. Compared with non-cancer controls, patients reported a higher prevalence of subjective cognitive impairment, especially in the subgroup of patients receiving chemotherapy. Regression analysis also confirmed that receipt of chemotherapy was one of the significant predictors of CRCI. Other risk factors related to CRCI in gynaecological cancer survivors included employment status and depression. Consistent with previous studies, the prevalence of CRCI was significantly higher in survivors with depression than in survivors without depression , 16, 34.While there is a growing concern regarding possible CRCI following primary cancer treatment , approprThis study found that patients after chemotherapy reported the lowest level of small-worldness index and global and local network efficiency, compared with age-matched non-cancer controls. Research evidence shows that disrupted structural networks have been demonstrated to have detrimental effects on cognitive functioning , 22, 38.Limitations of the cross-sectional study design were the inability to explore the course of CRCI over time; additionally, the study could not provide causal inferences for factors associated with subjective cognitive impairment. Thus, future prospective cohort studies should be conducted to explore the causal associations between CRCI and clinical factors, psychosocial variables, and brain networks. In addition, this study did not collect details about patients’ chemotherapy regimens. Hence, this study cannot discuss which chemotherapeutic agents may influence cognition. Finally, heterogeneity of clinical variables, such as cancer type, disease stage, and treatment modalities may have created sampling bias, which limits the generalisability of the results. While the present study has limitations that need to be addressed in future studies, its findings are important to communicate to patients and clinicians alike, especially due to the increasing level of concern about subjective cognitive impairment following cancer treatment.When compared with non-cancer controls, a considerable proportion of gynaecological cancer survivors may exhibit CRCI. After cancer treatment, 40.51% of Chinese gynaecological cancer patients perceived cognitive impairment. Moreover, cognitive impairment occurred not only in patients who had received chemotherapy, but also in approximately 10% of patients who were treated using surgery only. Lower cognitive functioning was associated with unemployment, receipt of chemotherapy, and depressive symptoms. This study provides the first evidence of brain structural network alteration in gynaecological cancer patients post-treatment, and offers novel insights regarding the neurobiological change mechanisms of CRCI in gynaecological cancer patients. Primary cancer treatment can result in a more random organisation of structural brain networks, which may reduce brain functional specificity and segregation and have implications for cognitive impairment. Future prospective and longitudinal studies are required to build upon the study findings in order to assess potentially relevant clinical and psychosocial variables and brain network measures, so as to more accurately understand the specific risk factors related to CRCI in the gynaecological cancer population. Such knowledge could inform the development of appropriate treatment and rehabilitation efforts to ameliorate CRCI in gynaecological cancer survivors.


Worldwide efforts to identify individuals infected with the hepatitis C virus (HCV) focus almost exclusively on community healthcare systems, thereby failing to reach high-risk populations and those with poor access to primary care. In the USA, community-based HCV testing policies and guidelines overlook correctional facilities, where HCV rates are believed to be as high as 40 percent. This is a missed opportunity: more than ten million Americans move through correctional facilities each year. Herein, the purpose of this paper is to examine HCV testing practices in the US correctional system, California and describe how universal opt-out HCV testing could expand early HCV detection, improve public health in correctional facilities and communities, and prove cost-effective over time.A commentary on the value of standardizing screening programs across facilities by mandating all facilities  to implement opt-out testing policies for all prisoners upon entry to the correctional facilities.Current variability in facility-level testing programs results in inconsistent testing levels across correctional facilities, and therefore makes estimating the actual number of HCV-infected adults in the USA difficult. The authors argue that universal opt-out testing policies ensure earlier diagnosis of HCV among a population most affected by the disease and is more cost-effective than selective testing policies.The commentary explores the current limitations of selective testing policies in correctional systems and provides recommendations and implications for public health and correctional organizations. Hepatitis C virus (HCV) infection is a global health issue of urgent importance. An estimated 170 million individuals are currently infected with HCV worldwide and result in 350,000-500,000 deaths a year. Globally, three to four million people are newly infected each year . And witPrison overcrowding, violence, and social isolation factor into prisoner initiation or continuation of behaviors that place them at risk for HCV infection (e.g. injection drug use) . Opt-out or universal testing policies test everyone and individuals may choose whether to participate. Data from state corrections system directors on HCV screening procedures at state prison systems reported that only 17 states (34 percent) had at least one state-prison facility offering routine opt-out HCV screening, with eight (16 percent) having no screening program to capture HCV data in place  , correctScreening for HCV in prisons is a public safety precaution. HCV-infected individuals re-entering society may act as bridging populations, transmitting infections to the community via resumed injection drug use upon release. Each year in the USA, approximately one of every three people infected with HCV is part of the correctional system. After release, most (95 percent) return to their community-of-origin. Early detection and engagement in care can prevent prisoners from unwittingly transmitting HCV to others after re-entering the general community. Universal opt-out HCV screening at entry into prison may result in a dramatic reduction in the number of new HCV infections over the next 30 years, benefits able to extend beyond the prison environment to the general community.This is a historic moment in the quest for HCV elimination thanks to the availability of novel, highly effective, therapy for HCV management. The recently available and highly effective oral direct-acting antivirals (DAAs) offer a cure for HCV-infected individuals. After treatment of one pill a day for 8-12 weeks, over 90 percent of patients reach undetectable viral levels and are “cured” of infection . CompareIn the following section, we provide a deeper look into the California State prison system, to illustrate the value of standardizing HCV screening programs across facilities by mandating all facilities  to implement opt-out HCV screening policies for all prisoners upon entry to correctional facilities.California state prison system is the largest prison system in the USA with 129,613 inmates in February 2017 . Based oThe successful implemented universal opt-out testing for HIV in California state prisons demonstrates the feasibility of expanded testing for HCV. In 2010 it led the USA, as the first state where all prison reception facilities tested every inmate for HIV, unless the individual declined . In respOpt-out HCV screening has been adopted for other US populations. Of the 37 California state correctional facilities, 11 are also reception centers for inmates entering prison with sentences of two or more years. Currently only three reception centers provide opt-out HCV screening to all prisoners upon entry to their facilities. Implementing universal HCV screening across all California prison facilities would provide a consistent standard to identify HCV positive inmates. Equitable access to testing can improve budgetary resources for care. Additionally, the HCV test would become part of the streamlined medical intake protocol and therefore less stigmatized.Require all facilities within a correctional system to offer opt-out HCV screening upon entry, ensuring equitable access to testing for all individuals moving through correctional facilities. This recommendation additionally meets the US legal requirement for facilities to provide health care to be “consistent with acceptable community standards” .Provide training for all medical intake staff on opt-out HCV screening and post-test disclosure counseling. Doing so will ensure a quality standard by which HCV screening is implemented. By packaging screening programs with test disclosure counseling, screening upon entry to correctional facilities will substantially decrease the number of individuals (estimated at 40-70 percent) who are unaware of their HCV status (Couple testing policies with needed services to protect against HCV transmission in correctional facilities. The majority of prisons lack harm reduction services to prevent and educate individuals. Improved identification of persons infected with HCV will allow evidence-based budgetary decisions for facilities to package testing with prevention, education, and care services.As the California universal opt-out HIV testing example illustrates, requiring all correctional facilities to provide opt-out HCV screening upon entry ensures more accurate HCV detection, equitable access to HCV screening, and streamlined systems for more efficient work environments.Universal opt-out screening offers several specific benefits. First, improved accuracy of recorded HCV rates in prisons. These data will inform evidence-based decision making for improved resource allocation by prison healthcare teams. Accurate estimates of the size of the HCV positive population can also help facilities assess the needs of their prison population. For facilities without a systematic screening program, the pool of infected individuals may be smaller than anticipated. By performing a one-time universal screening of all prisoners, the current HCV prevalence within a facility would be better understood. Second, early detection of HCV infections can improve HCV prevention and care. Equipping individuals with the knowledge of their HCV status upon release may help prepare parolees to access medical treatment and harm reduction programs in the community, potentially reducing transmission of HCV. Third, reduced stigma for individuals with or at risk for HCV, and subsequently reduce the risk of HCV transmission in prison and community populations. When people know they have HCV, they are less likely to engage in risky behaviors and are less likely to spread infection. Fourth, increase access to new HCV curative therapies. Delaying infection detection can delay or deny HCV care. Failure to treat may result in decompensated cirrhosis and hepatocellular carcinoma leading to high-intensity healthcare utilization, frequent hospitalizations and, potentially, liver transplantation. And fifth, offers a more cost-effective program, than voluntary or risk-based screening programs because HCV is treatable and curable. The advent of efficacious treatment with a high cure rate now makes wide-scale HCV screening an effective public health intervention.


Focal segmental glomerulosclerosis (FSGS) is the most common primary glomerular disease resulting in end-stage renal disease in the USA and is increasing in prevalence worldwide. It is a diverse clinical entity with idiopathic, genetic, metabolic, infectious, and other causes that culminate in a characteristic histologic pattern of injury. Proteinuria is a hallmark of FSGS as well as other primary and secondary glomerular disorders. The magnitude of proteinuria at disease onset and during treatment has prognostic implications for renal survival as well as associated cardiovascular morbidity and mortality. Significant advances over the last two decades have shed light on the molecular architecture of the glomerular filtration barrier. The podocyte is the target cell for injury in FSGS. A growing list of disease-causing gene mutations encoding proteins that regulate podocyte survival and homeostasis has been identified in FSGS patients. Several pathogenic and regulatory pathways have been uncovered that result in proteinuria in rodent models and human FSGS. The recurrence of proteinuria and FSGS after kidney transplantation is supporting evidence for the role of a circulating permeability factor in disease pathogenesis. These advances reviewed herein have significant implications for disease classification and therapeutic drug development for FSGS. The glomerular filtration barrier is composed of fenestrated endothelial cells, the glomerular basement membrane (GBM), and podocytes, terminally differentiated epithelial cells connected by interposed slit diaphragms between actin-rich foot processes . SeminalThis review discusses the knowledge to date of the underlying pathogenesis of podocyte injury leading to proteinuria and FSGS. We focus on the contributions of putative circulating factors, gene mutations, and viral infections.in vitro assay. Plasmapheresis was associated with a reduced proteinuria as well as a reduced glomerular permeability (The high recurrence rate of FSGS post transplant (estimated at 30–40%) suggests the presence of a circulating factor in recipients. A pivotal study by Savin et al. showed that plasma from patients with recurrent FSGS could increase the glomerular permeability to albumin in an eability . Gallon eability . The alleability . These s−/−) mice model, they showed that lipopolysaccharide (LPS)-induced proteinuria was uPAR-mediated, and uPAR upregulation following LPS stimulation led to αvβ3 integrin activation, with subsequent podocyte foot process effacement, increased podocyte motility, and proteinuria (in vitro and in vivo. In uPAR-knockout (Plaur−/−) mice, the supraphysiologic level of endogenous suPAR and exogenous suPAR injection induced αvβ3 integrin activation, resulting in podocyte injury and proteinuria. A recent study . To translate their findings to human disease, the investigators introduced whole peripheral blood mononuclear cells (human hematopoietic cells), drawn from individuals with recurrent FSGS and injected into NSG mice. This approach led to Gr-1lo cell expansion, suPAR upregulation, and proteinuria. However, CD34+-depleted PBMC from these patients failed to induce any proteinuria. This process was distinct from GVHD pathophysiology since T cell-depleted PBMC yielded similar results. Though human homologs of Gr-1lo − Sca-1 + myeloid progenitor cells have not been identified given the lack of Gr-1 or Sca-1 antigen, these findings implicate a common upstream pathway relevant to general CKD. The same investigators recently linked APOL1-risk alleles to the suPAR–β3 integrin activation pathway  and its soluble form (suPAR) have been the most studied , 17. uPAteinuria . The samnt study  has shed pathway . In thisThese findings have not been without controversy. Conflicting studies have shown that suPAR does not distinguish FSGS from other causes of nephrotic syndrome and that suPAR expression may be nonspecifically increased in the setting of a low eGFR –22. ThesProteomic analysis by liquid chromatography tandem mass spectrometry of recurrent FSGS plasma fractions that induce proteinuria in rats and enhance glomerular permeability to albumin led to the identification of CLCF1 –27. CLCFRecently, Delville et al.  screenedProteomic analysis of plasma and urine samples performed on patients with recurrent FSGS post transplant identified a high-molecular-weight form of Apolipoprotein A-I, named as ApoA-Ib in 93% of recurrent FSGS urines compared with <5% of those without recurrence, patients with non- FSGS proteinuric diseases, or patients transplanted for familial FSGS. Urinary ApoA-Ib had a sensitivity of 92.8% and a specificity of 98.1% for identifying FSGS relapse . UrinaryThere is evidence that plasma proteases may have a pathogenic role in recurrent FSGS. Vasodilator-stimulated phosphoprotein (VASP), a molecule involved in actin cytoskeleton organization is phosphorylated in response to exchange plasma from 10 patients with recurrent FSGS . VASP phBeginning with nephrin, the positional cloning of patients and relatives with familial FSGS has enabled the identification of numerous disease-causing podocyte genes Table . They enThe Rho family small GTPases  are essential in actin cytoskeletal dynamics, cell morphology, motility, and adhesion. While overactive RhoA can be deleterious, there is an emerging consensus that a relative predominance of RhoA activity relative to Rac1/Cdc42 favors a more stationary podocyte with intact foot processes , 54. ConThe onset of proteinuria is variable in genetic disease with autosomal-recessive etiologies generally manifesting clinically in childhood and autosomal-dominant traits in adulthood. Commercially available genetic testing is most likely to identify known disease-causing variants in infants and patients with familial and syndromic disease . Exomic Trypanosoma b. brucei. Over time, the parasite evolved into a serum resistance-associated protein (SRA) containing T. brucei rhodesiense that cause African sleeping sickness and capable of inactivating and evading Apol1. G1 and G2 Apol1 gene variants evade the SRA and are active against T. brucei rhodesiense. In this sense, this is analogous to malaria where the sickle trait is protective against parasitic infection but causes a hemoglobinopathy. Located on chromosome 22, the G1 and G2 alleles confer an increased risk for developing FSGS, hypertensive-associated kidney disease, and HIV nephropathy , p38 MAPK, and JNK. Interestingly, cytotoxicity was abrogated by SAPK inhibition and inhibition of K+ efflux  has been found to be critical for p53-induced oxidative cell injury with mTOR inhibition protecting against HIV-induced podocyte apoptosis . The proin situ hybridization techniques, parvovirus B19 DNA was localized at endothelial cells and both visceral and PECs. However, whether this viral infection results in podocyte dysfunction is still a matter of debate  remain unclear, representing an area of high unmet clinical need. Increasingly, podocyte-associated gene mutations are being identified in patients with FSGS. Expanded knowledge of the molecular architecture of the glomerular filtration barrier raises the prospect of a targeted therapeutic drug development.HIV and other viral infections can induce podocyte injury and FSGS with an increasing interest in the enhanced susceptibility conferred by APOL1-risk alleles. The numerous divergent etiologies and pathogenic mechanisms with resulting variable response rates to therapy and recurrence post transplant speak to the need for a reclassification of the disease into more clinically relevant subtypes. Further exploration of the contributions of JAK/STAT, Rho-GTPase, and ApoL1 to podocyte injury and survival could enhance the quest for novel therapeutic agents.YW, SS, and KC conceived the article contents, prepared the manuscript, and endorsed the final draft submitted.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.


In recent decades, bacteria’s therapeutic role has aroused attention in medicinal and pharmaceutical research. While bacteria are considered among the primary agents for causing cancer, recent research has shown intriguing results suggesting that bacteria can be effective agents for cancer treatment – they are the perfect vessels for targeted cancer therapy. Several bacterial strains/species have been discovered to possess inherent oncolytic potentials to invade and colonize solid tumors in vivo. The therapeutic strategy of using bacteria for treating cancer is considered to be effective; however, the severe side effects encountered during the treatment resulted in the abandonment of the therapy. State-of-the-art genetic engineering has been recently applied to bacteria therapy and resulted in a greater efficacy with minimum side effects. In addition, the anti-cancer potential of tumor-targeting bacteria through oral administration circumvents the use of the intravenous route and the associated adverse effects. This review aims to provide a comprehensive summary of the latest literature on the role of bacteria in cancer treatment. Despite the advancement in cancer treatment and detection, cancer has remained a major health problem and one of the leading causes of deaths worldwide. About 600,000 cancer deaths were projected to occur in the United States in 2016, out of 1.6 million newly reported cancer cases [The conventional chemotherapeutic agents used for the treatment of cancer possess non-specific toxicity toward normal body cells. Also the body cells that are exposed to chemotherapy often become resistant to drugs because of enhanced capability to repair DNA defects in cellular machinery which intervenes apoptosis. The exposed cells increase the production of enzymes that cause detoxification of drug and drug delivery services . These iHelicobacter pylori, which is associated with gastric cancer [Salmonella typhi which is associated with hepatobiliary carcinoma [Campylobacter Jejuni which is associated with small intestinal lymphomas [Chlamydia psittaci which is associated with ocular lymphomas [Mycobacte-rium tuberculosis which is associated with lung cancer [Citrobacter rodentium, which is associated with human colorectal cancer [Bacillus subtilis, competence stimulating peptide (CSP) from Streptococcus mitis and extracellular death factors (EDF) from Escherichia coli together with their tripeptide analogue, are reported to promote tumor cell invasion and angiogenesis through type I collagen extracellular matrix, which influences tumor metastasis [Bacteria are carcinogens and tumor promoters . Bacteric cancer , Salmonearcinoma , Campyloymphomas , Chlamydymphomas , Mycobacg cancer , and Citl cancer . In addil cancer , 12. Alstastasis . The quotastasis .Fig. 1ThClostridia, Bifidobacteria and Salmonella are capable of colonizing the hypoxic area of the tumor and destroy the tumor cells. Therefore, they are potential strains for selective tumor targeting therapy [Conversely, bacteria have shown great potential for cancer therapy. Bacteria of many species demonstrate the surprising ability to invade and colonize solid tumors, which often results in neoplasm growth retardation, and in some instances, complete tumor clearance . Differe therapy –21  has been demonstrated for centuries. However, the associated adverse side effects hinder its development. Adequate balance between the control of infection and the therapeutic benefit of bacteria is an essential requirement for a successful BMTT, but this can only be achieved via the heat-inactivation method . Recentlin urine . Such geAn important factor that applies to the spontaneous regression of cancer is the duality of the immune system . BacteriSalmonella typhimurium strain activates inflammasome pathways by damaging the signals released from cancer cells. This phenomenon significantly increases the amount of inflammatory cytokine IL-1β, TNF-α and Il-18 in tumors, which results in drastic tumor growth suppression [Salmonella typhimurium shows therapeutic efficacy for cancer through involving the inflammasome pathway [The ∆ppGpp pression . IL-1β ipression . The IL-pression . The canpression . Therefo pathway .E. Coli, which are capable of engulfing the solid tumors, are indirectly involved in clearance of some tumor cells (e.g. CT26) through infectious-defense mechanism. Once these bacteria invade the host, they stimulate the initiation of the defense mechanism of the host, which results in the production of lymphocytes T cells. The produced lymphocytes T cells are significantly involved in anti-tumor activity. During the induction phase of bacterial infection, CD8+ T cells are the only effectors responsible for tumor clearance; whereas, in the memory phase the clearance also involves CD8+ and CD4+T cells [+ T is reported to take part in the clearance of the original tumor after bacterial infection [+ and CD8+) have the potential to block the formation of a new set of tumors. The CD8+ T cell is said to have the additional ability to eradicate even the already established tumors [Anaerobic bacteria such as +T cells . CD8+ T nfection . Similard tumors . Furtherd tumors . Their rd tumors , 35. TheSalmonella enterica serovar Typhimurium to mice models indicated that haemorrhaging increases the flushing of bacteria into the solid tumor resulting in necrosis [The tumor necrosis factor (TNF- α) has the potential to damnify the vascular endothelial cells. TNF-α plays the significant part in the formation of the large haemorrhaging area that appears within the tumor. Systematic administering of necrosis . Moreovenecrosis . Thus, tSome substances secreted by bacteria, such as enzymes, can inhibit the growth of tumors. Several experimental studies discovered the therapeutic potential of different substances released from bacteria for treating cancer cell lines.Bacteriocins are cationic peptides that are synthesized by almost all groups of bacteria ribosomally. Bacteriocins are non-immunogenic, biodegradable and contain cancer cell-specific toxicities. The bacteriocins have the potential to serve as synergistic agents to conventional cancer drugs . Cancer Escherichia coli (E.coli), are known to have anti-cancer activities against a variety of human tumor cell lines in vitro, including breast cancer, colon cancer, bone cancer and uteri cell line HeLa  [Klebsiella pneumoniae, was found to induce apoptosis in some human malignant cell lines such as HeLa, Jurkat (T cell derived from acute T cell leukemia), RJ2.25 (a variant of Burkitt’s lymphoma), and colorectal carcinoma cells, with no effect on normal cells [Pediococcus acidilactici K2a2-3 was reported to have cytotoxic activities against HT29 (human colon adenocarcinoma) and HeLa cell lines [Lactobacillus lactis) possesses cytotoxic effect on MCF-7 (human breast adenocarcinoma cell line) [Colicins, the bacteriocin secreted from Enterobacteriaceae such as rcinoma) . Microcial cells . Pediocill lines . Similarll line) , HepG2 (ll line) , and HNSll line) , both inPseudomonas aeruginosa have shown anti-cancer activities [Pseudomonas aeruginosa strains [Furthermore, partially purified bacteriocins produced by certain bacteria such as tivities . Pyocin, strains –46, show strains . Likewis strains .Pseudomonus aeruginosa). The PDC phenazine was first isolated from Streptomyces species; it was demonstrated to be the potential agent for controlling metabolism and biofilm formation in Candida albicans [Multiple phenazine metabolites such as phenazine 1-carboxylic acid (PCA) and Phenazine 1,6-di-carboxylic acid (PDC) are derived from bacteria strains  are decisive in the pathogenesis of chronic infectious diseases [Salmonella tyhimurium and some other pathogens are known to cause severe haemorrhage within the tumor. Once the haemorrhae is activated, it induces the production of T cells that are very significant in biofilm induction [Biofilm is a primitive form of multicellular life that provides bacteria with tolerance strength against antibiotics and host defense mechanisms . Biofilmdiseases . Salmonenduction . Notwithnduction , recent Streptococcus agalactiae inhibit adhesion of cancer cells to endothelial cells, an essential step in cancer metastasis [Mariprofundus ferroxydans) as a new multifunctional drug carrier for cancer therapy and cancer hyperthmia.Anti-cancer drugs cause the induction of biofilm formation during cancer treatment, which results in metastasis distraction , 55. Simtastasis . Furthertastasis  demonstrWhile the above hypotheses avow for the potential of bacterial biofilm in cancer therapy, the evidence is relatively insufficient to build-up the case. However, the efficacy of bacteria biofilm for metastasis distraction calls for the further examination and investigation of the anti-cancer ability of bacteria biofilm.Apart from their direct anti-cancer effect, tumor-targeting bacteria can also be used as carriers for cancer therapeutic agents in cancer treatments. Recent studies have revealed that bacteria are capable of targeting both primary tumors and metastasis –63.Salmonella  and rhEndostatin in a murine model of malignant melanoma which resulted in the reduction of the tumor growth. The therapy is argued to be safer and effective, but also economically desirable – it decreases possible side effects and lowers the therapeutic expense. Moreover, Li et al. [Bifidobacterium adolescentis (non-pathogenic) as a vector for the expression of endostatin within tumors. Their findings showed that Bifidobacterium adolescentis strongly inhibit the angiogenesis and significantly inhibit local tumor growth. Bifidobacterium longum efficiently delivered the anti-angiogenic protein (endostatin) to murine liver tumors and induced anti-tumor activity [The growth and metastasis of solid tumors depend on the formation of new blood vessels (angiogenesis). Thus blocking tumor angiogenesis can be a reasonable approach to treat solid tumors. Jia et al.  applied i et al.  used Bifactivity , 67. In activity .The combined therapy of bacteriolytic and anti-angiogenic using tumor-targeting bacteria has also shown promising results. Bacteria are capable of invading the poorly perfuse tumor areas  using their unique metabolic features. They cause the inhibition of angiogenisis to kill residual tumor cells, and hence significantly increase the chance for tumor eradication . In addiEscherichia coli) expressing B18R enhanced the oncolytic potential of the vesicular stomatitis virus (VSVΔ51) to reduce tumor growth, and thus prolonged the survival of an aggressive tumor model.Oncolytic viruses (OVs) have shown positive outcomes for cancer treatment through their tumor-selective replication and multi-modality attack against cancers . The virbacteriorobot technique is a new innovative theranostic methodology of bacteria-based fabrication for tumor therapy. The technique uses bacteria as microactuators and microsensors to deliver microstructures for targeting and treating solid tumors [Salmonella typhimurium) in biocompatible/biodegradable alginate microbeads and attached flagellated bacteria  on the microbead to fabricate a bacteria-based microrobot in the targeted tumor region. The bacteria-encapsulated delivery system protects the bacteria from being attacked by the immune system, which is safer than the direct inoculation of bacteria [The d tumors . Variousd tumors , 77. In d tumors  encapsulbacteria .Bacteria are the double-edged sword in cancer therapy. Using bacteria for cancer therapy is feasible and its potential to treat solid tumors has been known for decades. However, the clinical application of this therapy never became routine because of the adverse uncontrollable side effects. In an effort to overcome these side effects, some attenuated species of bacteria capable of treating cancer have been recently identified and studied. These species of bacteria are considered safe for cancer therapeutic application with little or no side effects. While bacteria alone may not demonstrate fully therapeutic potential, their modifications as anti-tumor agents, anti-oncogenes or immunogenic antigens, and their combination with other therapeutic processes will improve their potential for cancer therapy. The arena of using bacteria as an anti-cancer agent is still new; further studies are imperative to scrutinize the clinical significance of bacteria-based cancer therapy. The findings presented in this review suggest that this promising cancer therapy needs to be optimized and developed further.