The function of p53 is best understood in response to genotoxic stress, but increasing evidence suggests that p53 also plays a key role in the regulation of metabolic homeostasis. p53 and its family members directly influence various metabolic pathways, enabling cells to respond to metabolic stress. These functions are likely to be important for restraining the development of cancer but could also have a profound effect on the development of metabolic diseases, including diabetes. A better understanding of the metabolic functions of p53 family members may aid in the identification of therapeutic targets and reveal novel uses for p53-modulating drugs. TP53 allele display an enormously increased cancer risk, a condition known as Li-Fraumeni syndrome  progress to irreversible senescence, a process named geroconversion, whereas cells in which mTOR is inhibited  ultimately achieve a reversible quiescent state . However, the majority of cancer cells display alterations in glucose metabolism  cells, pyruvate is subsequently fed into the mitochondrial tricarboxylic acid (TCA) cycle in order to generate NADH and FADHtabolism . Often, tabolism . Intermetabolism  but alsotabolism . Furthertabolism ; therefotabolism . Increastabolism . FurtherTP53-induced glycolysis and apoptosis regulator (TIGAR), which acts as a phosphatase that degrades F2,6BP and thereby decreases the activity of PFK1. Hence, p53, via TIGAR, lowers the glycolytic rate and would be predicted to promote the diversion of glycolytic intermediates into the PPP  takes place in the mitochondria and breaks down fatty acids into two-carbon units in order to yield acetyl-CoA, NADH, and FADHduce ATP . Fatty aduce ATP . Multiplduce ATP . Additioduce ATP , are knomulation .p53 plays a crucial role in lipid metabolism, participating in both normal and pathological conditions. True to its role as a tumor suppressor, p53 generally functions as a negative regulator of lipid synthesis by activating fatty acid oxidation and inhibiting fatty acid synthesis . Mouse eThe regulation by p53 of carbohydrate and lipid metabolism is tightly linked to another important function of p53: the regulation of ROS. In its role as a “guardian of the cell,” p53 can eliminate the deleterious effects of oxidative insult either by limiting ROS damage in cells that can be salvaged or by using ROS to eliminate cells damaged beyond repair. Hence, if the insult is transient and repairable, then p53 can activate a suite of antioxidant responses, many of which are linked to carbohydrate or lipid metabolism. For example, the activation of the PPP via TIGAR produces NADPH that can reduce glutathione, an important cellular antioxidant. Interestingly, a recent study showed that p53 can also directly promote GSH synthesis at the expense of nucleotide synthesis after serine deprivation, and, thereby, it actively controls ROS levels under conditions of metabolic stress . MitochoAutophagy (here referring to macroautophagy), which literally translates as “self-eating,” is an important cellular catabolic process carried out in the cytoplasm. Autophagy is characterized by the formation of double-membraned autophagosomes around cytoplasmic components that have been targeted for degradation. Once enclosed around their cargo, autophagosomes fuse with lysosomes, forming autolysosomes in which the delivered contents are then catabolized . AutophaThe relationship between p53 and autophagy is complex, given that p53 can both promote and inhibit autophagy in a context-dependent manner . This abDRAM-1 is activated by p53 and encodes multiple isoforms that regulate autophagy , thereby promoting cell-cycle arrest and survival in response to nutrient starvation (The activation of autophagy by (nuclear) p53 is well established and can occur through a variety of mechanisms. The mTORC1 pathway is a major inhibitory regulator of autophagy. Hence, by negatively regulating mTORC1 , p53 actutophagy . The proutophagy . Recentlutophagy . The tumutophagy . Recentlnditions , suggestnditions , ATG7 prarvation .−/− HCT116 cells also inhibited autophagy, the mutants showing the highest cytoplasmic localization having the greatest effect (The role of cytoplasmic p53 in the regulation of autophagy was established by suppressing p53 expression in colon cancer (HCT116) cells. Surprisingly, this resulted in the upregulation of autophagy, whereas the reintroduction of p53 repressed autophagy . In simit effect . It seemt effect . Under nt effect .Clearly, the identification of an ever-increasing number of functions for p53 provokes the important question of how the final response of the cell is determined. Many of the examples discussed above indicate that the particular outcome of the activation of p53 depends on the level of the insult. This may influence the activity of p53 itself as well as that of other proteins or transcriptional factors that ultimately determine the fate of the cell. p53 has clear relationships with other transcription factors that are known to be critical in the response to metabolic stress, most notably PGC, SREBP, and Nrf2. PGC-1α is a critical regulator of glucose, lipid, and energy metabolism, whose functions include the regulation of mitochondrial biogenesis and the expression of antioxidants. Under conditions of mild metabolic stress, p53 has been shown to increase PGC-1α expression, resulting in the induction of an antioxidant response . MoreoveTp63 and Tp73 genes are transcribed from two distinct promoters, resulting in either full-length proteins that retain a full transactivation (TA) domain (TAp63 and TAp73) or N-terminally truncated isoforms (ΔNp63 and ΔNp73) that lack part of this domain but still retain some ability to activate transcription. In addition, alternative RNA splicing results in different C termini for both p63 and p73 isoforms, termed α-ε and α-η, respectively. TAp63 and TAp73 can supplement p53 function by transactivating p53 target genes, resulting in cell-cycle arrest and apoptotic cell death, although p63 and p73 also have distinct transcriptional targets. In contrast, ΔNp63 and ΔNp73 generally have antiapoptotic functions and have been shown to act as dominant negatives for inhibiting the function of p53 family members. However, ΔN isoforms also activate specific sets of target genes and function independently of other p53 family isoforms. Despite these similarities and overlap in activity, p63 and p73 each have functions that are strikingly different from those of p53, and both proteins play critical roles in development . Unlike p63, p73 is essential for proper neural development, and p73 knockout mice display developmental defects in the CNS. Analysis of selective TAp73 or ΔNp73 knockdown suggested that ΔNp73 isoforms are necessary for neuronal survival, whereas TAp73 is required for the long-term maintenance and differentiation of neuronal stem cells. Both p63 and p73 are important for germ cell maintenance. p63 controls the quality of the female germline by eliminating damaged oocytes, whereas p73 maintains genomic stability of the oocyte pool.p53 belongs to a family of transcription factors that also includes p63 and p73, functional homologs of p53 that show high sequential and structural similarity. Both the Sirt1, AMPKα2, and LKB1 and thereby coordinates fat and glucose metabolism. Loss of TAp63 results in defects in lipid utilization, fatty acid synthesis, and FAO, and mice lacking TAp63 consequently display insulin resistance as well as symptoms of obesity, type 2 diabetes, and premature aging  had a shortened lifespan  and increased levels of ROS . Furtherem cells . Another+/m mice . HoweverThe metabolic functions of p53 are emerging as critical not only for tumor suppression but also for maintaining normal cellular homeostasis. Nevertheless, many questions remain. Clearly, different stress signals can activate different p53 responses, as measured by transcriptional profiles, and several proteins that contribute to the ability of p53 to promote adaptation to metabolic stress are preferentially activated by p53 in response to nutrient deprivation . However


In this study we aimed at characterizing the regulation of hepatic metabolic pathways by the p53 transcription factor.Analysis of gene expression following alteration of p53 status in several human- and mouse-derived cells using microarray analysis, quantitative real-time PCR, chromation immunoprecipitation and reporter gene assays. A functional assay was performed to determine lipid transfer activity.plfp, abca12 and cel) was further characterized. In addition to HepG2, the genes were induced following activation of p53 in human primary hepatic cells isolated from liver donors, p53-dependent regulation of these genes was evident in other cell types namely Hep3B cells and mouse hepatocytes. Furthermore, p53 was found to bind to the genes’ promoters in designated p53 responsive elements and thereby increase its transcription. Importantly, p53 augmented the activity of secreted PLTP, which plays a major role in lipoprotein biology and atherosclerosis pathology.We identified a novel role for the p53 protein in regulating lipid and lipoprotein metabolism, a process not yet conceived as related to p53, which is known mainly in its tumor suppressive functions. We revealed a group of 341 genes whose expression was induced by p53 in the liver-derived cell line HepG2. Twenty of these genes encode proteins involved in many aspects of lipid homeostasis. The mode of regulation of three representative genes (These findings expose another facet of p53 functions unrelated to tumor suppression and render it a novel regulator of hepatic lipid metabolism and consequently of systemic lipid homeostasis and atherosclerosis development.


The human waking EEG spectrum shows high heritability and stability and, despite maturational cortical changes, high test-retest reliability in children and teens. These phenomena have also been shown to be region specific. We examined the stability of the morphology of the wake EEG spectrum in children aged 11 to 13 years recorded over weekly intervals and assessed whether the waking EEG spectrum in children may also be trait-like. Three minutes of eyes open and three minutes of eyes closed waking EEG was recorded in 22 healthy children once a week for three consecutive weeks. Eyes open and closed EEG power density spectra were calculated for two central (C3LM and C4LM) and two occipital (O1LM and O2LM) derivations. A hierarchical cluster analysis was performed to determine whether the morphology of the waking EEG spectrum between 1 and 20 Hz is trait-like. We also examined the stability of the alpha peak using an ANOVA.The morphology of the EEG spectrum recorded from central derivations was highly stable and unique to an individual (correctly classified in 85% of participants), while the EEG recorded from occipital derivations, while stable, was much less unique across individuals (correctly classified in 42% of participants). Furthermore, our analysis revealed an increase in alpha peak height concurrent with a decline in the frequency of the alpha peak across weeks for occipital derivations. No changes in either measure were observed in the central derivations.Our results indicate that across weekly recordings, power spectra at central derivations exhibit more “trait-like” characteristics than occipital derivations. These results may be relevant for future studies searching for links between phenotypes, such as psychiatric diagnoses, and the underlying genes  by suggesting that such studies should make use of more anterior rather than posterior EEG derivations. The EEG can be recorded non-invasively and inexpensively, with accurate estimates of cortical oscillations made with just a few minutes of recording. EEG recordings are of great utility not only for learning about the healthy brain, but also reflect changes in brain structure/function associated with psychiatric disorder. The heritability and stability of the EEG add to its utility. For example, studies of twins have found that the human waking EEG is one of the most heritable traits with heritability estimates ranging between 0.55 and 0.9,2, depenIn addition to being highly heritable, the waking EEG spectrum is stable across time. A number of studies in adults, adolescents, and children have shown high test-retest reliability between sessions  for both absolute and relative spectra,9-11. Mientire waking EEG spectrum in early adolescents (ages 11 to 13 years) over weekly intervals and examine the extent to which the waking EEG spectrum is a biological trait in this age range. We used a cluster analysis based on distance between the three weekly-recorded power density spectra to assess whether the EEG spectrum is trait-like. Compared to ICC analysis, in cluster analysis the entire EEG spectrum is taken into account, making it more representative of an individual, thus enhancing its utility and validity. Unlike ICC, cluster analysis does not rely on a priori knowledge of repeated measures within a given individual and thus does not make use of intra and inter-individual variability. Rather, in cluster analysis, spectra are grouped according to their similarity. Furthermore, we examined whether trait-like characteristics, as determined by the cluster analysis, vary in central and occipital EEG derivations. In order to compare our results with previous stability studies, we also examined the stability of the alpha peak across recordings using an ANOVA.The aim of the current study was to examine the stability of the Twenty-two healthy right-handed early adolescents, aged 11 to 13 years  participated in this study. All participants were pre/early pubertal (n = 13) or mid-pubertal (n = 8) with the exception of one female who was late pubertal.Participants were recruited using television and newspaper advertisements, as well as at schools, and at a meeting of the association for parents with gifted children. The Zurich cantonal ethical committee (KEK) for research on human participants approved the protocol and the participants’ legal guardian gave written informed consent. This study conforms with the Code of Ethics of the World Medical Association (Declaration of Helsinki), printed in the British Medical Journal (18 July 1964). Participants were compensated for their participation by cinema, book or CD vouchers and a T-shirt.Participants were healthy, had no history of neurologic and psychiatric disease, and were medication free. For three days before each experimental session participants were asked to abstain from caffeine and medication and to adhere to a regular sleep–wake schedule . Wrist-worn actimeters and sleep logs verified compliance. The data used in the current study is the control data from another study.Participants came to the laboratory on three different occasions in weekly intervals, always at the same time of the day. At each session, a baseline waking EEG was recorded to assess the stability of waking EEG in early adolescents over weekly intervals.EEG , electrooculogram (EOG), and electrocardiogram (ECG) were recorded with a polygraphic amplifier Artisan . The analog signals were high-pass  and low-pass filtered (−3 dB at 67.2 Hz), sampled at 256 Hz, and recorded using Rembrandt DataLab .Waking EEG was recorded continuously for 6 min . Participants sat on a chair, rested their head on a chin rest, and were instructed to avoid movement. Vigilance was ensured by continuous online visual inspection of the recordings and alerting the subjects via intercom when signs of drowsiness were present.Artifacts were visually marked and artifact-free segments were subjected to spectral analysis  using MATLAB . All participants had at least thirty 2-s epochs of artifact free data in each condition  which were used to calculate the power density spectra. Frequencies between 1 and 20 Hz were analyzed.2 test to assess whether the rate of correct clustering was significantly different between derivations or ‘eyes open/closed’ conditions.Hierarchical cluster analysis based on Euclidean distance was used to examine whether the log-transformed EEG spectra from 1 to 20 Hz) were trait-like using MATLAB functions PDIST and LINKAGE (average aggregation strategy). Cluster analysis was performed in several steps. First, a power spectrum was obtained for each recording/participant and represented as a vector consisting of power density at all frequency bins . Next, the distance between all vectors  was calculated and vectors with small distances between them were clustered together, while vectors that were far apart were clustered separately. Finally, the distance between vectors is represented visually as a dendrogram, which consists of upside-down U-shape lines where the height of the U represents the distance between the connected objects  of the alpha peak. The alpha peak was determined on an individual basis because significant interindividual variability in the frequency of the alpha peak has been shown[To be able to compare our results with previous studies and investigate whether the characteristics of the alpha rhythm vary across sessions and derivations, we determined the frequency and height  visual analog scale similar to Aitken, was admIn order to compare the morphology of the entire spectrum we performed a hierarchical cluster analysis (see Methods). The number of participants that clustered successfully over all three recordings was dependent on EEG derivation Figure . For the2(1) = 13.54, p = 0.0002; Eyes open: χ2(1) = 9.40, p = 0.002). To demonstrate this difference, four participants whose spectra clustered in the eyes closed condition for derivation C3LM but not O1LM are shown in Figure 2(1) = 0.23, p = 0.63, Eyes open: χ2(1) = 0, p = 1) or left and right occipital (Eyes closed: χ2(1) = 2.28, p = 0.13, Eyes open: χ2(1) = 0, p = 1) derivations  and frequency (Hz) of the alpha peak in the waking EEG spectrum using an ANOVA with factor ‘week’ . Average power spectra are shown in Figure We examined the stability of the peak height  = 7.86; p = 0.001; O2LM: F = 7.134, p = 0.006). Peak height increased between weeks one and two (O1LM: t(21) = −2.68; p = 0.014; O2LM: t(21) = −2.78; p = 0.011) and between weeks one and three (O1LM: t(21) = −4.12; p <0.001; O2LM: t(21) = −2.9; p = 0.009). Means and standard deviations for peak height and frequency are shown in Table A change in mental tension across weeks was observed (mean (SD): week 1 = 30.3 (16.8); week 2 = 29.7 (24.2); week 3 = 27.6 (21.1); F = 4.248; p = 0.036). A post-hoc paired t-test revealed a decline in mental tension between weeks one and three (t(21) = 2.37; p = 0.028) but not between weeks one and two (t(21) = 1.88; p = 0.075). No other variables showed a significant change across weeks.We included mood as a covariate in the ANOVA analysis in order to examine whether the change in mental tension had an impact on the observed effects in the occipital derivations. Examining the data in this way, there was no longer an effect of week for O1LM peak frequency in the eyes closed condition and O2LM peak height in the eyes open condition.The current study used two different measures to examine the stability of the EEG spectrum in early adolescents: a cluster analysis approach and characterization of the alpha peak. Overall, the results showed that the waking EEG is highly stable across recordings and unique to an individual. In addition, we were able to demonstrate that the degree to which the EEG spectrum is trait-like is dependent on the brain region.entire spectrum rather than limited to the alpha band. Although the alpha oscillation comprises an important cortical rhythm, functional neuroanatomy is more accurately reflected in the entire EEG spectrum.The current study used cluster analysis to examine whether the waking EEG in early adolescents represents a trait. Thus, the unique contribution of our analysis is not solely the examination of stability, but also the quantification of interindividual variability. Another advantage of the current analysis is that clustering is based on the interindividual variability rather than low intraindividual stability may account for the lower rate of clustering in occipital versus central derivations. For example, greater variability across individuals was observed in the frequency and shape of the waking EEG spectrum in central compared to occipital derivations , we would not expect successful clustering for any participant by chance. We do not expect that the difference between central and occipital derivations is due to less stability of the alpha peak in the occipital regions because our ANOVA analysis revealed a change in alpha peak characteristics in only the eyes open condition, whereas the difference between occipital and central derivations was present in both eyes open and closed conditions. The lack of In a previous study of trait-like characteristics of the sleep EEG in a different sample of adolescents, Tarokh et al. found thThe alpha frequency band is an important rhythm to consider when examining the heritability and stability of the waking EEG. Several studies have shown that alpha peak power is a highly stable EEG parameter and thatSeveral limitations of this study are important to note. With respect to our analysis of the stability of the alpha frequency peak, our frequency resolution was 0.5 Hz, which limits our ability to detect more subtle changes in frequency across recording session. Furthermore, this analysis was restricted to four derivations, which limits our ability to examine regional differences in further detail. We examined a narrow age range and the degree to which the regional differences we observe are specific to this developmental stage is unknown. In fact, there are large differences in the maturational trajectory of different cortical regions. Future We used a novel method to show not only that the waking EEG spectrum is trait-like, but that the degree to which the EEG is trait-like depends on brain region. This finding has implications for resting state waking EEG studies in search of biological markers of cognitive capabilities and psychiatric disorders suggesting that such studies should use central rather than occipital derivations.The authors have no competing interests to declare.SPL and PA designed the study. LT, SPL and DB performed data analysis. DB and SPL participated in data collection. LT, SPL, DB and PA wrote the manuscript. All authors read and approved the final manuscript.Peter Achermann, Sarah P Loughran Shared senior authorship.Dominik C Benz, Leila Tarokh Shared first authorship.


Posttraumatic stress disorder (PTSD) and major depressive disorder (MDD) frequently co-occur after traumatic experiences and share neurocognitive disturbances in verbal memory and executive functioning. However, few attempts have been made to systematically assess the role of a comorbid MDD diagnosis in neuropsychological studies in PTSD.The purpose of the current study is to investigate neurocognitive deficits in PTSD patients with and without MDD. We hypothesized that PTSD patients with comorbid MDD (PTSD+MDD) would have significantly lower performance on measures of verbal memory and executive functioning than PTSD patients without MDD (PTSD–MDD).n=84) and patients with PTSD–MDD (n=56).Participants included in this study were 140 treatment-seeking outpatients who had a diagnosis of PTSD after various single traumatic events and participated in a randomized controlled trial comparing different treatment types. Baseline neuropsychological data were compared between patients with PTSD+MDD (The PTSD+MDD patients had more severe verbal memory deficits in learning and retrieving words than patients with PTSD alone. There were no differences between the groups in recall of a coherent paragraph, recognition, shifting of attention, and cognitive interference.The results of this study suggest that a more impaired neurocognitive profile may be associated with the presence of comorbid MDD, with medium-sized group differences for verbal memory but not for executive functioning. From a clinical standpoint, being aware that certain verbal memory functions are more restricted in patients with comorbid PTSD and MDD may be relevant for treatment outcome of trauma-focused psychotherapy. Posttraumatic stress disorder (PTSD) and major depressive disorder (MDD) are common outcomes after experiencing a traumatic event , as well as consistent deficits in verbal memory  a PTSD diagnosis according to DSM-IV; 2) a single traumatic event that led to the development of PTSD and had stopped at the time of inclusion; 3) age between 18 and 65 years; 4) mastery of the Dutch language. Exclusion criteria were: 1) acute suicidality; 2) current severe MDD or current severe alcohol or substance dependence according to DSM-IV; 3) a lifetime psychotic disorder according to DSM-IV; and 4) a severe personality disorder according to the SCID-II screener , 1 (= rarely), 3 (= sometimes), and 5 (= often) and the total PTSD score (range 0–110) consists of the sum of the scores. The depression scale of the Hospital Anxiety and Depression Scale (HADS) was used to measure the severity of the depressive symptoms by self-report  is a test to measure shift of attention, planning and cognitive flexibility Reitan, . PatientThe Stroop Color Word Test is thought to measure selective attention and cognitive flexibility . If this overall test was significant, we examined group differences on the separate test variables within the univariate general linear model. Two-tailed tests were applied throughout and level of significance was set at α=0.05 for the multivariate tests. Level of significance for the univariate analyses was set at α=0.01 to correct for multiple testing. Partial-eta squared was calculated as an effect size for significant differences.Analyses were conducted using SPSS version 19.0 . Chi-square tests and independent Demographic and clinical characteristics of the two groups are displayed in F-test revealed a significant difference between the groups for the CVLT, F=2.89, p=0.011. Univariate analyses revealed that the PTSD+MDD group scored significantly lower on CVLT sum of trials 1–5, F=8.63, p=0.004, η2=0.058; CVLT short-term cued recall, F=8.15, p=0.005, η2=0.057; CVLT long-term free recall, F=10.98, p=0.001, η2=0.069; and CVLT long-term cued recall, F=7.77, p=0.006, η2=0.053, than the PTSD–MDD group. Multivariate F-tests did not reveal significant differences between the groups for the RBMT, F=2.90, p=0.059; TMT, F=1.10, p=0.337; or Stroop, F=1.88, p=0.137.Mean scores and standard deviations of the two groups on the neurocognitive tests are displayed in The main finding of the current study is that treatment-seeking patients with PTSD and comorbid MDD have more pronounced verbal memory deficits than patients with PTSD alone, with medium-sized differences between the groups. These verbal memory deficits in the group with PTSD and MDD seem to be restricted to the encoding, short-term retrieval and long-term retrieval of separate words, since there were no significant differences in retrieval of a coherent paragraph or in recognition between the two groups. Various aspects of executive functioning in the present study were similar for both groups as well, as expressed by a lack of differences on measures of mental speed, shift of attention, selective attention, and cognitive interference. In sum, encoding and retrieval of separate words was a bigger challenge for PTSD patients who had comorbid MDD diagnoses, than for those who had PTSD without MDD. Results of this study thus suggest a somewhat more impaired neurocognitive profile for PTSD patients with comorbid MDD.Our results confirm and extend the studies of Sachinvala et al.  and JohnWe note that the group with PTSD and comorbid MDD in our study also had more severe PTSD symptoms than the group without comorbid MDD. Therefore, it is possible that the differences we found between the groups are partly attributable to greater PTSD symptom severity. Moreover, it could be the case that PTSD symptom severity contributes to the other neuropsychological processes  for which we were not able to confirm statistically significant group differences. However, more severe PTSD symptoms and a comorbid MDD diagnosis are such intertwined constructs that it is very difficult, and may not even be clinically meaningful, to statistically separate the influence of these two variables (Miller & Chapman, Limitations of this study include that we did not administer tests for all types of attention and that we had to exclude patients with severe MDD, as this was an exclusion criterion for the treatment trial. It would be interesting to examine especially sustained attention in samples with PTSD+MDD versus PTSD alone, as depressive symptoms were also found to play a role in sustained attention in PTSD (Meewisse et al., In conclusion, patients with PTSD and comorbid MDD seem to have more difficulty in learning separate units of verbal information and retrieving this information in the short and long term than PTSD patients without comorbid MDD, even when the severity of the MDD diagnosis is only mild to moderate. PTSD symptom severity may contribute to these difficulties. From a clinical standpoint, it is good to be aware that these aspects of verbal memory are more restricted in patients with comorbid PTSD and MDD, as verbal memory performance can influence the treatment outcome of trauma-focused psychotherapy (Wild & Gur,


This tumour suppressor protein has three main domains; the N-terminal transactivation domain, the CTD  and the core domain (p53C) that constitutes the sequence-specific DBD (DNA-binding region). Most p53 mutations related to cancer development are found in the DBD. Aggregation of p53 into amyloid oligomers and fibrils has been shown. Moreover, amyloid aggregates of both the mutant and WT (wild-type) forms of p53 were detected in tumour tissues. We propose that if p53 aggregation occurred, it would be a crucial aspect of cancer development, as p53 would lose its WT functions in an aggregated state. Mutant p53 can also exert a dominant-negative regulatory effect on WT p53. Herein, we discuss the dominant-negative effect in light of p53 aggregation and the fact that amyloid-like mutant p53 can convert WT p53 into more aggregated species, leading into gain of function in addition to the loss of tumour suppressor function. In summary, the results obtained in the last decade indicate that cancer may have characteristics in common with amyloidogenic and prion diseases. Disruption of the p53 network usually has severe consequences that favour cell survival and tumour progression –3. p53 iThe human p53 protein comprises 393 amino acid residues and three main functional regions: the N-terminal activation domain, which is able to interact with a variety of proteins; the CTD, responsible for tetramerization; and the core domain (p53C) comprising residues 94–312 that constitute the sequence-specific DBD (DNA-binding region) of the protein  Figure . More thTP53) are frequently associated with increased susceptibility to cancer development. Inactivation of p53-regulated pathways has been described in over 50% of all human cancers, making them interesting targets for cancer therapies  is located at 17p13.1 and is composed of 11 exons  breast cancer  reduction assay  are found in most proteins. These small APRs are normally sheltered from aggregation due to intra-protein interactions or because they are hidden in the hydrophobic protein core. However, when the protein structure is disturbed by mutations or environmental changes, these sequences can be exposed, thus favouring protein aggregation . One of lymphoma . This coThe formation and kinetics of WT and mutant p53 amyloid oligomers and fibrils has been widely discussed ,79–82. pThe DNA-binding properties of p53 are frequently lost upon p53 aggregation; however, the small cognate double-stranded DNA has been shown to stabilize both the p53C domain and full-length p53 and to have the potential to rescue aggregated and misfolded species  Figure . TherefoThe presence of intermediate states that participate in the p53 aggregation route has been highlighted in different studies ,72,86. MTP53 gene proposes that the presence of even a single mutant protein in the formation of the p53 tetramer would lead to loss-of-function of p53 [The dominant-negative hypothesis for WT p53 inactivation in cells carrying a mutation in one of the alleles of the n of p53 ,88. Our n of p53 ,16. In on of p53  , conjugated ubiquitin and several cell cycle-regulating cyclins, among other molecules [In addition to self-aggregation and combined mutant-WT aggregation, p53 aggregation has also been described where other proteins function as a platform. This is the case for the above mentioned p53–Hsp70–Mdm2 interaction  and alsoolecules . p53 wasolecules .We conclude that aggregation of p53 into a mixture of oligomers and fibrils sequesters the native protein into an inactive conformation that is typical of a prionoid. Our findings that amyloid aggregates are present in biopsies of breast cancer tissues, especially in the aggressive tumours, show the relevance of this prion behaviour in cancer pathogenesis ,76. In fThe prion-like behaviour of oncogenic p53 mutants, as represented in


The tumor suppressor gene p53 is involved in a variety of cellular activities such as cellular stress responses, cell cycle regulation and differentiation. In our previous studies we have shown p53’s transcription activating role to be important in osteoblast differentiation. There is still a debate in the literature as to whether p53 inhibits or promotes differentiation. We have found p53 heterozygous mice to show a p53 dependency on some bone marker gene expression that is absent in knockout mice. Mice heterozygous for p53 also show a higher incidence of osteosarcomas than p53 knockout mice. This suggests that p53 is able to modify the environment within osteoblasts. In this study we compare changes in gene expression resulting after either a transient or stable reduction in p53. Accordingly we reduced p53 levels transiently and stably in C2C12 cells, which are capable of both myoblast and osteoblast differentiation, and compared the changes in gene expression of candidate genes regulated by the p53 pathway. Using a PCR array to assay for p53 target genes, we have found different expression profiles when comparing stable versus transient knockdown of p53. As expected, several genes with profound changes after transient p53 loss were related to apoptosis and cell cycle regulation. In contrast, stable p53 loss produced a greater change in MyoD and other transcription factors with tissue specific roles, suggesting that long term loss of p53 affects tissue homeostasis to a greater degree than changes resulting from acute loss of p53. These differences in gene expression were validated by measuring promoter activity of different pathway specific genes involved in differentiation. These studies suggest that an important role for p53 is context dependent, with a stable reduction in p53 expression affecting normal tissue physiology more than acute loss of p53.  In adgulation ,7. In ougulation . Other sgulation ,10. Whilgulation . In embrgulation . So whilThe importance of tissue specificity with regard to p53 function is further supported by studies carried out in p53 transgenic mice ,14. In aSince p53 responds to multiple stimuli and can affect multiple, different cellular processes, changes in p53 expression will affect cells differently depending on whether the change is transient or stable. In this study we test this by using shRNA interference to reduce p53 levels in a bipotential cell line, C2C12, transiently and compared the changes in gene expression of genes involved in p53 pathways with a stable reduction of p53. We found greater changes in expression of MyoD and of other transcription factors with tissue specific roles in the stable p53 knockdown cells, which suggests that long term partial loss of p53 expression affects tissue homeostasis to a greater degree than changes resulting from transient loss of p53. Additionally, we show differences in the promoter activities of genes involved in differentiation between stable and transient loss of p53. Together this points toward the role of p53 as being context dependent, with a stable loss affecting cellular differentiation more than a transient loss.2/95% air.  In order to study the role of p53 in the differentiation of mesenchymal stem cells, the bi-potential cell line C2C12  was selected because it can differentiate into myotubes and can be induced to osteoblasts, depending on the culture media . These cThe p53 HuSH-29 shRNA plasmids , were used to create transient and stable knockdown lines of p53 within the C2C12 cells. We used three different shRNA sequences to knockdown p53. Control cells received in a similar sized non effective scrambled sequence (TR20003). SuperFect  was used for both transient and stable transfections according to manufacturer’s protocol. Additionally, for obtaining stable clones, cells were selected post transfection using puromycin. Single cell clones were expanded, passaged and characterized before use. To determine the changes at the transcriptional level, RNA was isolated either using TRI Reagent  or the QIAGEN RNeasy Mini Kit (cat. no. 74104) prep system depending on the assay used. Genomic contamination of the RNA was removed using the Ambion DNA Free kit . DNA free RNA was quantified using the Nanodrop2000 .Semiquantitative RT-PCR was used to measure the relative expression of p53 after shRNA introduction as well as, the various bone and muscle markers.. RNA was reverse-transcribed using the QIAGEN OneStep RT-PCR kit (cat. no. 210210) using equivalent amounts of RNA. The following primers were ordered from QIAGEN, and used according to their protocol: Gapdh (cat. no. QT01658692) and p53 (cat. no. QT00101906). To measure the intensity of the resulting PCR, the samples were run on a 1% agarose gel containing ethidium bromide and the image captured on a Kodak system. The image was then quantified using the UnscanIt™ software and the relative quantification was calculated using Gapdh to normalize the expression.2 First Strand Synthesis Kit  according to the standard protocol. A master mix using the RT2 SYBR Green/ROX kit  was made for each sample and then the array was then loaded and ran on an ABI Prism 7300 Real Time PCR System, using the settings as defined by SABiosciences for their arrays. Once run, the data was normalized using the included genes on the array, then fold changes were calculated by using the standard 2-ΔΔCt method, using the array of the control cells as the second Δ.In addition to looking at the various differentiation markers, a more in depth analysis of pathway perturbations was also assayed using the QIAGEN/SABiosciences p53 Signaling Pathway PCR Array . Genes included on the array are direct targets of p53, regulators of p53, or genes in a p53 pathway that are downstream of p53, and cover many of the different pathways p53 is found to affect, such as apoptosis, differentiation, and cell cycle regulation, as well as control genes for normalization, genomic DNA contaminating and RT efficiency. The RNA obtained for a stable line of shRNA plasmid 34 (S34), 36 (S36), as well as, a transient transfection created from plasmid 36 (T36) and a control line of C2C12 cells was made using QIAGEN RNeasy Mini Kit (cat. no. 74104) prep was first quantified using the Nanodrop 2000, then 1.5 µg of total RNA was reverse-transcribed using the RT5’-GTTCTTCACGCCCAAAAGATG-3’, reverse 5’-GGACAGTTGGGAAGAGTGTCATT-3’), Pten , p300 , Rb1 , Bag1 , Casp2 , Casp9 , Mcl1, , Numb , and Prkca , with 18S  used as a normalizer. These primers were used in an assay using SYBR Green QPCR on duplicate test samples. Samples of RNA free of DNA contamination were reverse transcribed using the Ambion High Capacity cDNA kit . A master mix containing the Applied Biosystems PowerSYBR® Green 2x Master Mix , an equal amount of the cDNA reaction, and water to make 25 µl volume samples were created for each gene. The mix was then aliquoted into a 96 well plate, primer added, and ran on an Agilent 7300 Real Time PCR System. Fold changes were calculated using the 2-ΔΔCt method using 18S as the first Δ with the control samples as the second Δ.In order to validate results of the e PCR arrays, several genes that underwent changes in expression were selected, and quantitative real-time PCR (QPCR) was performed (see Results section) on triplicate samples of S36, T36, and control C2C12 cells. Specific primers for Realtime analyses are as follows. p53 , MyoD (Integrated DNA Technologies (IDT), Coralville, IA, forward C2C12 clones with the different p53 shRNAs were grown to confluency for stable lines or two days post transfection for transient transfected lines. Total protein was isolated from each plate using M-PER mammalian protein extraction reagent  with supplemented Roche Complete ULTRA Tablets protease inhibitor . Protein concentration was measured using Bradford assay . Following isolation and quantification, the proteins were subjected to western blot analyses. A 25 or 50 μg fraction of each protein extract was separated on a SDS PAGE gel, then electro transferred onto presoaked PVDF paper . The membranes were then blocked with 5% milk protein in PBS/T, and the primary antibody for either p53  or β-actin  was added and allowed to incubate overnight at 4°C. The membrane was then washed with PBS/T three times, and HRP conjugated secondary antibodies to mouse or rabbit IgGs  was added, washed again, and exposed using the ECL Western blotting reagents . The image was then captured on a Kodak system and UnscanIt™ was used to obtain quantification.Several gene specific reporter assays were carried out to examine the effect of transient and stable p53 knockdown. Our reporter constructs consisted of mainly luciferase reporters but some of them were Chloramphenicol Acetyl Transferase (CAT) based reporters. Specific details for both assays are provided below. When stable p53 knockdown lines were used, cells received just the reporter construct. When the assay was carried out in cells with transient knockdown of p53, we either used a specific shRNA construct or a mixture of three p53 shRNA constructs. Specific details are provided in figure legends. Control cells received a scrambled sequence of a similar length in the same plasmid backbone (for knockdown of p53) and/or the empty vector (reporter assays). Forty-eight hours after transfection, they were trypsinized, pelleted, and then resuspended in serum free media. An aliquot was used to count cells for normalization, the rest was assayed using Promega’s Bright GloTM Luciferase Assay Kit . Equal volumes of the resuspended cells and Bright-GloTM were mixed and incubated for 2 minutes. A Turner Designs TD20/20 Luminometer was used to record the luciferase activity for each sample. Samples were read for 10 seconds, each after a two second delay. Luciferase readings for each of the samples were then normalized according to the number of cells/ml. All measurements were carried out on triplicate samples and experiments were repeated at least thrice.In order to evaluate the functional activity of p53 we transfected cells with, pG13-luc , a plasm14C Chloramphenicol. The product was extracted with xylene and radioactivity measured using a liquid scintillation counter. All measurements were carried out on triplicate samples and experiments were repeated at least thrice.Forty-eight hours after transfection the cells were lysed and equal amounts of protein were used to measure chloramphenicol acetyltransferase (CAT) activity using n-Butyryl CoA and Because p53 is involved in the development and differentiation of C2C12 cells into later, more committed cell types, the promoters of certain genes representing various pathways were assayed for activity in the stable and transient p53 knockdowns to test to see if transient or stable loss of p53 had different effects upon the expression of bone or muscle markers. The notch pathway was assayed using the Hey1-luc plasmid . In addi< 0.05 for significance. To visualized the difference between how T36 and S36 differ when compared to control cells, clustering was done using Cluster3 [All statistics were done in GraphPad Prism 5 (Microsoft), using either unpaired t-tests or one-way ANOVA with Tuckey correction for multiple testing and post hoc tests, and requiring a p Cluster3  by usingCluster3 , which wIn order to create a stable knockdown of p53, we isolated puromycin resistant single cell clones after transfecting various p53 shRNA constructs in a plasmid conferring puromycin resistance into C2C12 cells. Individual clones were expanded and characterized. The same constructs were also used for transient reduction of p53, but no selection was performed and cells were harvested after 48 hours for RNA and protein. Our intention was to generate cells with a similar knockdown in p53 using the same shRNA construct. In order to rule out possible dosage dependent effects between and among the different constructs and type of tranfections, several methods were used to assess the knockdown Therefore in addition to determining extent of p53 knockdown at the transcriptional and translational level, we tested the level of p53 functional activity using a construct containing 13 copies of a p53 binding sequence . This coIn order to look for more global differences between acute loss, from the transient knockdown, and a long term loss of p53 the SABioscience Q-PCR array was used to assay the changes in genes related to p53. We chose to compare RNA from stable and transient knockdown created with the shRNA (S36 vs. T36). RNA from both these cells was first compared to control cells receiving a scrambled shRNA sequence. As expected, many of the targets showed changes with loss of p53 expression. Hierarchical clustering of the expression changes between the two lines are shown as compared to control cells. Two types of change were observed one where genes are up-regulated in the acute loss compared to the long term loss of p53, and a second cluster with genes up-regulated in long term p53 loss when compared to the acute loss. . As expeSome of our previous studies demonstrated that the amount of p53 within osteoblasts might affect cell proliferation and differentiation variably -28. SincSeveral genes belonging to apoptosis and differentiation related pathways showing alterations in the array were chosen for validation using realtime PCR. For this analysis we created new stable lines with p53 knockdown using the same shRNA. Transient knockdown of p53 was also carried out in triplicate. RNA from these newly generated cell lines were compared to results obtained from the gene array. As shown in < 0.001; In addition to the expression data, we wanted to determine if specific pathways related to differentiation are activated differently dependent on whether p53 loss was stable or transient. The notch pathway is known to be important for tissue differentiation and has been shown to be important for both myoblast and osteoblast differentiation ,30. When< 0.001; < 0.001; < 0.01; Since the C2C12 cells can differentiate into either myocytes or osteoblasts, we tested the promoter activity of the genes Muscle Creatine Kinase (MCK), a late marker of myogenic differentiation, and osteocalcin, a late marker of osteoblast differentiation. When p53 is stably reduced the loss results in a significant decrease in activity of the osteocalcin promoter (p p53 is a transcription factor that regulates a number of different genes affecting cellular conditions ranging from proliferation, apoptosis, and DNA repair to cellular metabolism and senescence. While a role for p53 in cellular differentiation is well accepted, there is still a debate on whether p53 facilitates or opposes differentiation. This stems from the fact that loss of p53 during development is associated with little or no effect on tissue differentiation . In the In an attempt to separate the different roles of p53 we studied the short and long term effect of p53 loss on its target genes. In this study we utilized C2C12, a bipotential cell line capable of differentiation towards myoblast and osteoblast, to determine the effect of transient and stable knockdown of p53 using RNA interference. An acute loss of p53 lowered the mRNA expression for a number of genes related to apoptosis. This is expected, as one of the major roles of p53 is in regulating apoptosis, a function that represents an immediate response to prevent damaged DNA and or cells from survival. However, these same genes show a small increase in expression after long term loss of p53; perhaps as an adaptation to reduced p53 levels. Specifically, members of the caspase family of genes, Casp2 and Casp9, as well as the Casp2 assembly protein Cradd, are all down-regulated with an acute loss of p53, whereas they appear to be up-regulated when p53 is reduced for the long term. It is also possible that the change seen in caspases with stable p53 loss may reflect other functions unrelated to apoptosis as has been seen for some cell types -40. SimiThe apoptotic pathway is not the only pathway that appears to differ according to the length of p53 knockdown. Genes involved in p53 related differentiation pathways varied in expression based on the type of p53 loss. MyoD and Numb are well known differentiation and cell fate related genes ,43. ValiWe studied lineage specific, as well as, lineage independent signaling pathways that might be affected by the knockdown of p53 in C2C12 cells. We investigated activity of the Notch pathway and did not find differences between stable and transient knockdown as in both cases this pathway was depressed with loss of p53. In a genome wide study carried out to determine p53 regulators, the notch pathway, especially Hey1, was found to be members of an evolutionarily conserved network governing p53 function . It was These studies indicate that while the role of p53 is important to protect cells from damage by regulating key apoptotic genes, p53 has an additional role in maintaining tissue homeostasis. This may be especially important in osteoblasts since bone remodels throughout life. The presence of p53 may be critical to allow for orderly progression of the different steps of the differentiation process. p53 is a direct regulator of matrix proteins osteopontin and osteocalcin ,50. Oste


AbstractHemiptera: Coccoidea) known from Hungary has increased in the last 10 years by 39 (16.6 %), to a total of 274 species belonging to 112 genera in10 families. The family Pseudococcidae is the most species rich, with 101 species in 34 genera; Diaspididae contains 59 species in 27 genera; Coccidae contains 54 species in 27 genera; and the Eriococcidae contains 33 species in 8 genera. The other 6 coccoid families each contain only a few species: Asterolecaniidae (7 species in 3 genera); Ortheziidae (7 species in 4 genera); Margarodidaesensu lato (5 species in 5 genera); Cryptococcidae (3 species in 2 genera); Kermesidae (4 species in 1genus); and Cerococcidae (1 species). Of the species in the check list, 224 were found in outdoor conditions, while 50 species occurred only in indoor conditions. This paper contains 22 species recorded for the first time in the Hungarian fauna.The number of scale insect species ( Hemiptera: Coccoidea) live on a wide variety of plant species and many of them are important agricultural pests. Publication of new knowledge of this insect group is therefore very important from a practical viewpoint. The distribution data of different species may serve also as a reliable biodiversity indicator in different terPageBreakritories, such as nature reserves and agricultural or urban landscapes. The distribution data may also reflect the progress of climatic changes , Coccidae (54 species) and Eriococcidae with 33 species. The new species to the Hungarian fauna recorded here belong to the Pseudococcidae, Diaspididae and Eriococcidae. Most of the species in the checklist  are native and live outdoors. The check list contains 50 introduced  species, mainly occurring indoors in Hungary on ornamental plants in greenhouses and buildings. Of these indoor species, 33 occurred only in greenhouses or buildings  and 7 were found exclusively on imported tropical/subtropical fruits for consumption. Four of the species living in greenhouses sometimes also occur outdoors. Four other species, which are typically found on imported fruit, also appear in greenhouses from time to time. Two of the newly recorded species were found on imported nursery plant material. In the present list, 22 species are new to the Hungarian fauna. According to these data, Hungary is the most scale-insect-species-rich country in in Central Europe  in the last ten years, and currently totals 274 species in ten families  and 2. Tl Europe .PageBreakPageBreakPageBreakPageBreakPageBreakPageBreakPageBreakPageBreak106 (38.69%) species are considered as widely distributed Pan-Palaearctic species, 75 (27.37%) are widely distributed Euro-Siberian species, 91 (33.21%) are cosmopolitan, and only two species are known to originate from the Mediterranean subregion.No species should be considered as truly endemic only on the basis of its presence in a checklist, because the lack of a species in the surrounding countries is most likely due to inadequate exploration of those areas . Out of PageBreakperate climate, but with several submediterranean, xerophilous habitats. In addition, the Great Hungarian Plain belongs to the steppic province of the Palaearctic Region, which ranges from Hungary to China and the Far East , Aspidiotus destructor Signoret, 1869, Aulacaspis yatsumatsui Takagi, 1977). The following species have become significant pests in Hungary in recent years: Aspidiotus nerii Bouché, 1833; Coccus hesperidum Linnaeus, 1758; Planococcus citri ; Pseudococcus longispinus ; Pseudococcus viburni  and Saissetia coffeae   .Pseudococcus elisae Borchsenius, 1957). The number of species in this category is surprisingly low, compared to the number of pests living on various fruits exported from the different regions of production. The low species number reflects the efforts made by exporting countries to prevent the spread of invasive pests. It should be noted that most of these species were unable to establish in Hungary even indoors in greenhouses or on ornamental plants in buildings, despite repeated introductions over more than one hundred years. On the other hand, some of them have become regular pests in Hungary, which has lead to some overlap with the category in Aspidiotus nerii, Planococcus citri and Pseudococcus viburni occur in greenhouses and buildings, while Pseudaulacaspis pentagona and Carulaspis caruelii are found outdoors.A detailed study of the scale insects introduced into Hungary on tropical and subtropical fruits was published by PageBreakPageBreakPageBreak


As a hallmark of tumor cells, metabolic alterations play a critical role in tumor development and could be targeted for tumor therapy. Tumor suppressor p53 plays a central role in tumor prevention. As a transcription factor, p53 mainly exerts its function in tumor suppression through its transcriptional regulation of its target genes to initiate various cellular responses. Cell cycle arrest, apoptosis and senescence are most well-understood functions of p53, and are traditionally accepted as the major mechanisms for p53 in tumor suppression. Recent studies have revealed a novel function of p53 in regulation of cellular metabolism. p53 regulates mitochondrial oxidative phosphorylation, glycolysis, glutamine metabolism, lipid metabolism, and antioxidant defense. Through the regulation of these metabolic processes, p53 maintains the homeostasis of cellular metabolism and redox balance in cells, which contributes significantly to the role of p53 as a tumor suppressor. Further understanding of the role and molecular mechanism of p53 in cellular metabolism could lead to the identification of novel targets and development of novel strategies for tumor therapy. Tumor suppressor p53 plays a central role in tumor prevention -4. p53 iAs a transcription factor, p53 mainly exerts its function in tumor suppression through transcriptional regulation of its target genes -4. In re18flurodeoxyglucose than normal tissues. Remarkably, recent research has begun to answer these questions. These studies have led to the concept that the metabolic alterations are a hallmark of tumor cells . Th. Th63]. p53 can further regulate glycolysis through its regulation of PI3K/AKT and mTOR signaling pathways Figure . The abeIn addition to glucose metabolism, p53 regulates many other aspects of cellular metabolism, including glutaminolysis and fatty acid oxidation Figure . GLS2 acEmerging evidence also shows that p53 is involved in lipid metabolism. In response to nutritional starvation, p53 induces fatty acid oxidation to drive the TCA cycle to provide energy for cells Figure . Guanidi+ dependent enzyme in mitochondrial matrix, which catalyzes proline degradation in mitochondria and thereby reduces intracellular ROS levels [Oxidative stress and increased levels of ROS in cells play an important role in tumorigenesis. Recent studies have shown that reducing the ROS levels and enhancing antioxidant defense is an important mechanism of p53 in tumor suppression. Mitochondrial oxidative phosphorylation is a main source of endogenous ROS in cells. Whereas p53 promotes mitochondrial oxidative phosphorylation to maintain the homeostasis of cellular energy metabolism, p53 also plays a fundamental role in reducing the intracellular ROS levels mainly resulted from mitochondrial oxidative phosphorylation and maintaining the redox balance. To exert its antioxidant function, p53 induces a group of antioxidant genes, including sestrins 1/2, TIGAR, GPX1, ALDH4, GLS2, and Parkin, especially under conditions of nonstress or low stress, to lower ROS levels and prevent DNA damage (Figure S levels . By incrS levels -53. In aS levels . As a trS levels ,19.Interestingly, in addition to the function of antioxidant defense, p53 can exert prooxidant function through transcriptional up-regulation of a group of prooxidant genes depending on the levels of oxidative stress that cells are facing (Figure Here we reviewed our current understanding of the role and mechanisms of p53 in maintaining the homeostasis of cellular energy metabolism and the redox balance in cells. Whereas cell-cycle arrest, apoptosis, and senescence are traditionally accepted as the major mechanisms by which p53 exerts its tumor suppressive function, these findings provide strong evidence that the functions of p53 in maintaining the homeostasis of cellular energy metabolism and the redox balance in cells contribute significantly to the role of p53 as a tumor suppressor.Metabolic alterations are a hallmark of cancer cells. Ample evidence has shown that the metabolic alterations are critical for the growth, proliferation and survival of tumor cells. Therapeutic strategies targeting cancer metabolism are being developed and tested for cancer treatment. For instance, Metformin, an anti-diabetic drug, which regulates cell metabolism and the AMPK/mTOR signaling, has been already selected as a candidate for tumor therapy ,86. ReceThe authors declare that they have no competing interests.YL and JL prepared the initial draft of the paper. ZF modified and finalized the paper. All authors read and approved the final manuscript.


MYL3, encoding the essential light chain of myosin, are rare and have been associated with sudden death. Both recessive and dominant patterns of inheritance have been suggested. We studied a large family with a 38-year-old asymptomatic HCM-affected male referred because of a murmur. The patient had HCM with left ventricular hypertrophy (max WT 21 mm), a resting left ventricular outflow gradient of 36 mm Hg, and left atrial dilation (54 mm). Genotyping revealed heterozygosity for a novel missense mutation, p.V79I, in MYL3. The mutation was not found in 300 controls, and the patient had no mutations in 10 sarcomere genes. Cascade screening revealed a further nine heterozygote mutation carriers, three of whom had ECG and/or echocardiographic abnormalities but did not fulfil diagnostic criteria for HCM. The penetrance, if we consider this borderline HCM the phenotype of the p.V79I mutation, was 40%, but the mean age of the nonpenetrant mutation carriers is 15, while the mean age of the penetrant mutation carriers is 47. The mutation affects a conserved valine replacing it with a larger isoleucine residue in the region of contact between the light chain and the myosin lever arm. In conclusion, MYL3 mutations can present with low expressivity and late onset.Hypertrophic cardiomyopathy (HCM) is caused by mutations in genes encoding sarcomere proteins. Mutations in  MYL3 which encodes the myosin essential light chain (ELC) of the sarcomere . It. ItMYL3 MYL3 mutation p.V79I. The mutation segregates with disease and is absent in 600 control alleles in a family in which no other mutation was identified in known HCM-associated genes. The phenotypic presentation of the p.V79I mutation is that of classical HCM with asymmetric septal hypertrophy, just as previously described for the p.A57G mutation in two unrelated Korean families [We have described a patient with mild HCM associated with a novel, dominantly inherited, families , 17 but families , p.M149Vfamilies . ExpressThe part of beta myosin containing the IQ1 motif that interacts with the N-C-loop of ELC harbours a number of HCM-associated mutations, that are, p.S782D , p.S782NHomology modelling suggests that the p.V79I mutation may interfere with the interaction between ELC and myosin heavy chain, a mechanism which is believed to be the cause of disease for a number of reported mutations. Based on this evidence we find it likely that the p.V79I mutation is disease causing. However, it can not be ruled out that the mutation may only be associated with hypertrophy when other triggering conditions are present. In this case, the proband is very obese; obesity has recently been associated with a cardiac hypertrophic response in mice fed a high-fat diet through inactivation of the Foxo3a transcription factor via the Akt pathway . As the MYL3 have been associated with sudden death [MYL3-based genetic predisposition, the hypertrophic phenotype and obesity in the proband should also strengthen the recommendation to the proband to lose weight.The finding of a clinically silent mutation with low expressivity and late onset raises the question of whether it should entail a detailed followup of mutation carriers. However, as most mutations in en death , it woul


A significant amount of evidence suggests that the p38-mitogen-activated protein kinase (MAPK) signalling cascade plays a crucial role in synaptic plasticity and in neurodegenerative diseases. In this review we will discuss the cellular localisation and activation of p38 MAPK and the recent advances on the molecular and cellular mechanisms of its substrates: MAPKAPK 2 (MK2) and tau protein. In particular we will focus our attention on the understanding of the p38 MAPK-MK2 and p38 MAPK-tau activation axis in controlling neuroinflammation, actin remodelling and tau hyperphosphorylation, processes that are thought to be involved in normal ageing as well as in neurodegenerative diseases. We will also give some insight into how elucidating the precise role of p38 MAPK-MK2 and p38 MAPK-tau signalling cascades may help to identify novel therapeutic targets to slow down the symptoms observed in neurodegenerative diseases such as Alzheimer's and Parkinson's disease. The MAPKs are a specific class of serine/threonine kinases which respond to extracellular signals such as growth factors, mitogens, and cellular stress and mediate proliferation, differentiation, and cell survival in mammalian cells. There are 4 distinct groups of MAPKs within mammalian cells: the extracellular signal-related kinases (ERKs), the c-jun N-terminal kinases (JNKs), the atypical MAPKs , and the p38 MAPKs . The p38in vivo. In this paper we will especially focus our attention on the role of two p38 MAPK substrates in neurons: MAPK-activated protein kinase 2  and tau protein. The involvement of the p38 MAPK-MK2 and p38 MAPK-tau signalling cascades in neuroinflammation, actin remodeling, and tau hyperphosphorylation in neurodegenerative diseases will also be discussed. Highlighting the functional role of specific p38 MAPK substrates in neurodegenerative disease will be of particular importance as these could be potential signalling targets which could be exploited therapeutically to slow cognitive decline occurring in normal ageing and in neurodegenerative disease.In the present paper we will give an overview of p38 MAPK localisation, activation, and the functional role of this signalling cascade in the mammalian brain, especially the activation of the p38 MAPK cascade during synaptic plasticity in the hippocampus. Although p38 MAPK isoforms have been shown to be highly expressed in the brain, only a handful of brain-specific substrates for p38 MAPK have been characterised in vitro experiments in 293T HEK cells showed a similar distribution profile for both the endogenous and exogenous MKK3 and MKK6 proteins in that they are both nuclear and cytoplasmically localised within the cell . It. Itα/β Mammation , 62. In ytokines . This siβ plaques or NFTs formed from free aggregated neuronal tau within the brain. However the relationship between these structures and the symptoms of cognitive impairment and memory loss that is associated with the disease remains uncertain. Increasing evidence has shown that the stability of dendritic spines and actin remodelling may participate in the pathology of the disease. The loss of synapses is a common occurrence within postmortem tissue of AD patients , 83β is  neurons , contribal cells . In AD, deposits  through  in vivo . Upon thin vitro. It has been demonstrated that release of proinflammatory cytokine IL-1 from activated microglia increased the levels of tau phosphorylation in neurons. These changes are partly mediated through activation of p38 MAPK as a significant increase in the levels of phospho-p38 MAPK was observed upon application of IL-1β in cultures of neocortical neurons and microglia [in vitro that IL-1β-induced tau phosphorylation was considerably decreased in neuronal culture [Glial-neuron interactions and the effect these interactions have on tau phosphorylation have been analysed icroglia . Additio culture , again hα has been seen to localise to areas where NFTs, amyloid plaques, and glial cells are present both within human AD brain and transgenic mouse models [β. This complex, which forms through Aβ-peptide-induced dimerisation, suggests a connection between the aberrant processing of APP and the ASK-1-MKK6-p38 MAPK cascade which is involved in inflammation and abnormal tau phosphorylation [In vitro activation of MKK6-p38 MAPK has led to tau phosphorylation at specific sites, the most efficient being Ser-396, which has been suggested to have a functional role in microtubule binding. Abnormal phosphorylation at Ser-396 is observed in AD brain but not in normal functioning adult brain [β peptide, can activate the upstream MKKK ASK-1 and MKK6, consequently activating p38 MAPK, which can directly phosphorylate tau protein hence linking deposition of Aβ plaques to downstream tau phosphorylation through the activation of the p38 MAPK signalling cascade under pathophysiological conditions. Under pathophysiological conditions, activated p38e models , 89, 90.e models  and to be models , the prerylation . In vitrlt brain . Furtherlt brain . This suPick's disease is another severe neurodegenerative disorder, which involves progressive dementia and aphasia through the development of Pick bodies, which are comprised of neurofibrils formed of aggregated phosphorylated tau. It is known that oxidative stress is involved in instigating Pick's disease, and since it has been highlighted that the p38 MAPK cascade is activated upon such stimuli, it may play an important role in this disease as well. It has been observed in post-mortem brain tissue that phosphorylated p38 MAPK localises to the Pick bodies which contain highly phosphorylated tau protein, and since p38 MAPK is capable of phosphorylating tau, as described above, it emphasises the importance of p38 MAPK in this disease as well as AD and other related tauopathies .α-synuclein (SNCA), which is a protein present within the LBs of PD patients and known to play a pivotal role in the development of the disease. α-Synuclein is highly expressed in neuronal tissue and is subject to many post-translational modifications such as phosphorylation and ubiquitination, although its physiological function is poorly characterised. It has been suggested, however, that these post-translational modifications participate in neurotoxicity [α-synuclein is the predominant fibrillar component of the proteinaceous Lewy bodies seen in PD. Additionally, it has been observed that amplified α-synuclein levels are linked to an increase in neuroinflammation as it has been shown that α-synuclein activates p38 MAPK and other MAPKs in glial cells, a finding that is supported by the fact that extracellular α-synuclein released from damaged neurons interacts with microglia [α, which can induce and promote neuroinflammation as a physiological neuroprotective mechanism. If the glial cells become overactivated, however, as may occur through elevated levels of extracellular α-synuclein released from damaged Lewy body containing neurons, then chronic inflammation could be established and lead to neuronal cell death. Parkinson's disease is the second most prevalent neurodegenerative disease and around 127,000 people in the UK are currently living with the disease, which has been estimated to rise by 28% by the year 2020 . PD invotoxicity  as aggreicroglia . As descα, thus causing neuroinflammation and potentially neuronal cell death. An investigation by Culbert et al. [MAPKAP kinase-2, one of p38 MAPK more prevalent substrates has also been implicated within PD, where it has been shown that MK2-deficient mice show decreased levels of neuroinflammation and loss of dopaminergic neurons within the substantia nigra after treatment with the Parkinson's inducing neurotoxin MPTP compared to MK2 wild-type mice . MK2, aft et al.  demonstrConsiderable progress has been made in the understanding of the functional role of the p38 MAPK signalling cascade in synaptic plasticity in the hippocampus and its potential role in neurodegenerative diseases such as AD. However, less is known regarding the role of the direct targets of p38 MAPK, such as MK2 and tau, in regulating neuroinflammation and the actin cytoskeleton in dendritic spines of neuronal cells. Growing evidence suggests that remodelling of actin at dendritic spines plays a crucial role in synaptic plasticity and therefore in cognitive processes such as learning and memory. Furthermore, recent findings in animal models have linked early symptoms of AD with loss of cognitive functions, combined with a reduced number of dendritic spines in the hippocampus. Abnormal dendritic spine morphology has also been observed in brain tissue from patients suffering from AD. Therefore, with an ageing population continuing to grow and the consequent rise in AD, elucidating the precise role of p38 MAPK-MK2 and p38 MAPK-tau signalling cascades in controlling actin remodelling becomes very important as it may identify novel targets to slow down the cognitive decline observed in normal ageing and in the early stages of neurodegenerative diseases.


The transcription factor p73 belongs to the p53 family of tumour suppressors and similar to other family members, transcribed as different isoforms with opposing pro- and anti-apoptotic functions. Unlike p53, p73 mutations are extremely rare in cancers. Instead, the pro-apoptotic activities of transcriptionally active p73 isoforms are commonly inhibited by over-expression of the dominant negative p73 isoforms. Therefore the relative ratio of different p73 isoforms is critical for the cellular response to a chemotherapeutic agent. Here, we analysed the expression of N-terminal p73 isoforms in cell lines and mouse tissues. Our data showed that the transcriptionally competent TAp73 isoform is abundantly expressed in cancer cell lines compared to the dominant negative ΔNp73 isoform. Interestingly, we detected higher levels of ΔNp73 in some mouse tissues, suggesting that ΔNp73 may have a physiological role in these tissues. The Trp73 gene belongs to the p53 family of transcription factors and, like the other members, is transcribed into different isoforms -4. TP73


TP53, which occur in 96% of serous ovarian tumors, alter the core molecular pathways involved in drug response. One subtype of TP53 mutations, widely termed gain-of-function (GOF) mutations, surprisingly converts this protein from a tumor suppressor to an oncogene. We term the resulting change an oncomorphism. In this review, we discuss particular TP53 mutations, including known oncomorphic properties of the resulting mutant p53 proteins. For example, several different oncomorphic mutations have been reported, but each mutation acts in a distinct manner and has a different effect on tumor progression and chemoresistance. An understanding of the pathological pathways altered by each mutation is necessary in order to design appropriate drug interventions for patients suffering from this deadly disease.Ovarian cancer is the most lethal gynecological malignancy, with an alarmingly poor prognosis attributed to late detection and chemoresistance. Initially, most tumors respond to chemotherapy but eventually relapse due to the development of drug resistance. Currently, there are no biological markers that can be used to predict patient response to chemotherapy. However, it is clear that mutations in the tumor suppressor gene  TP53, in 96% of all serous ovarian tumors. TP53 encodes p53, a tumor suppressor that acts as a major control hub for the cellular response to various stresses, including DNA damaging chemotherapy. Once activated, p53 protects against cancer by functioning as a sequence-specific transcription factor or through protein: protein interactions, activating cell cycle arrest, apoptosis, and DNA damage repair.One of the main goals of clinical research is to establish better drug regimens for patients suffering from specific diseases. In the field of ovarian cancer research, the current standard therapy of platinum and paclitaxel has been in place without change for almost twenty years . The lacRB1 or APC that are largely inactivated by mutations that result in deletion or truncation .,50.49,50R273 are the most common TP53 alterations in ovarian carcinomas, with the most frequent mutations resulting in an amino acid change to histidine or cysteine. Mutation of R273 significantly alters the target DNA sequence without causing structural distortions, thus maintaining the ability to bind DNA [In vivo studies in mice with an engineered R273H mutation in the endogenous locus (p53R273H/−) demonstrate no difference in survival as compared to p53−/− mice [p53R273H/− mice develop more carcinomas, including high numbers of lung adenocarcinomas and squamous cell carcinomas. Additionally, cells cultured from these tumors demonstrate an increased capacity to proliferate [TP53 null background results in resistance to cisplatin [Mutations in codon bind DNA ,76. In v−/− mice . Howeverliferate . In studisplatin ,41. It risplatin .et al. showed that after DNA damage, R273H/C p53 complexes with the transcription factor NF-Y on the promoters of the cell cycle genes cyclin A, cyclin B, cdk1, and cdc25c and activates their expression [The mechanisms underlying increased resistance to some chemotherapeutic agents likely involve new protein interactions as well as new transcriptional targets ,61,77,78pression ,79. Genepression . These ipression . Whetheri.e., NF-Y and SP-1), although the interaction has the opposite effect compared to oncomorphic p53 [In addition to mediating chemoresistance, interactions between R273H/C mutant p53 and other proteins may contribute to oncogenesis or tumor progression. The best characterized R273H p53 interaction is with its family member p63. Specifically, p63 activity is inhibited by R273H p53 binding, which in turn promotes TGF-β-induced metastasis . The losphic p53 ,69,83. TR248 is the second most commonly altered amino acid in ovarian carcinomas, occurring at a frequency of 6.02% [R248W or R248Q). This alteration does not significantly affect the overall conformation of the p53 protein but changes the DNA binding response element, thereby altering interactions with DNA [248 can increase oncogenicity through increased invasion and chemoresistance to particular drugs. In lung cancer cells, although there are no differences in the proliferation rate of clones stably expressing this mutant, cells with this mutation do display significantly higher migration [P-glycoprotein) [R248W TP53 knock in mice (p53R248W/−), expression of R248W p53 markedly accelerates the development of lymphomas and sarcomas [TP53 R248Q mutant was created, and these mice display accelerated tumor onset as well as a shortened survival [The codon of 6.02% ,34. The with DNA . Cellulaigration . The mecprotein) . As withprotein) . The difsarcomas . In addisurvival .R248 mutations in chemoresistance, disruption of this complex sensitizes cells to adriamycin and cisplatin and induces apoptosis. These data also highlight that, while mutation at R248 is widely considered to predict for resistance to therapy, patients must also be screened for the SNP at codon 72 to achieve an accurate prediction for response.The p53 mutant R248 can activate the transcription of several genes associated with chemoresistance, namely c-Myc, CXCL1, PCNA, ABCB1 (MDR1), and IGF1R ,77,86,87TP53 mutation in all cancers and is the best characterized mutation. Many different types of cancer cell lines that harbor this mutation as well as murine cancer models have been used to study the oncogenic properties of R175H, and these studies have suggested a role in chemoresistance [R273H: greater tumorigenic potential with enhanced proliferative capacity [p53−/− mice [R175H TP53 transgenic mice display increased genomic instability. In experiments in lung cancer cells devoid of p53, overexpression of the oncomorph R175H p53 enhances resistance to both etoposide and cisplatin [The p53 oncomorph R175H is the third most common sistance . Transgecapacity . Moreove−/− mice ,90. In aisplatin . Furtherisplatin .Similar to the R273 and R248 p53 oncomorphs, the R175H mutated protein can interact with proteins NF-Y, p63, p73, and SP-1 to increase cell cycle progression, survival, metastasis, and chemoresistance ,79,81,82Y220C mutation occurs in 3.74% of ovarian carcinomas, though it has not been as extensively studied as the other TP53 mutations. Though it is not defined by others as an oncomorphic mutation, one might speculate it has oncomorphic activity given its reported interaction with p63 and p73 and its transcriptional activation of stathmin, which, as a microtubule destabilizer, is implicated in chemoresistance in many solid tumors [The d tumors ,96.Y220C mutation. Specifically, the mutated codon is not in direct contact with DNA but rather creates a binding pocket in the core domain of the protein [in vivo cell or mouse models of cancer. Thus, the binding pocket presents an opportunity for a novel strategy of cancer treatment based on the idea of restoring WT p53 function and thereby sensitivity to DNA damaging chemotherapy.In contrast to the lack of understanding of its functional properties, several structural studies have provided novel insight into the  protein  that is  protein . ImportaTP53 mutation has context-specific effects. Additionally, each oncomorphic mutation may have differing actions in response to different cancer therapeutics used. Experiments should be performed to compare the various oncomorphic mutants in the same parental cancer cell line in order to reduce variability. Two main strategies should be utilized for these studies: over-expression of oncomorphic mutants in p53-null cells, as well as knockdown of WT p53 concomitant with re-expression of the oncomorphic p53 variants  since clearly all TP53 mutants are not equal. Ovarian cancer is an ideal model to study p53 mutant biology because mutations in this gene are so common. Based on our calculations, oncomorphic mutations may comprise up to 20% of all mutations in ovarian tumors. However, a single therapy may not be adequate for all tumors with oncomorphic TP53 mutations. Extensive research in vitro as well as in vivo in mouse models is necessary to examine the biology of each mutation, particularly those that occur with high frequency. The studies described above will pave the way for appropriate comparisons of specific mutants, and possibly identify common signaling pathways that can be targeted for anti-cancer drugs. For example, exciting new methodologies may help progress the field of mutant p53 research by attempting to find oncomorphic p53 binding consensus sequences, as well as transcriptome sequencing to identify oncomorphic p53-associated biomarkers that can be used as predictive tools for resistance to chemotherapy. It will be further necessary to complement in vitro studies with animal models of ovarian cancer to understand the function of these oncomorphs and identify drug strategies that will be most useful.The process of carcinogenesis involves gaining oncogenic activities as well as losing tumor suppressive activities. Tumors that acquire an oncomorphic TP53 oncomorphic mutations can lead to identifying better therapies for patients with oncomorphic mutations in their tumors, and potentially avoid the high incidence of chemoresistance that is currently one of the barriers to improving survival for this deadly disease.As new technologies helps advance methodologies used to study mutant p53 biology, the most useful information will be discovering commonalities between the mutants. Understanding the basic science of in vitro and in vivo studies as well as several clinical trials showing that restoration of WT p53 function causes rapid tumor regression in mice and prolonged survival in humans [TP53 gene into tumors using an adenovirus. Excitingly, 50% (8 of 16) women with recurrent ovarian, peritoneal, or fallopian tube cancer that were treated with the replication-deficient adenovirus encoding human recombinant WT TP53 (SCH 58500) showed a decrease in serum CA125 levels, indicative of clinical response, with minimal side effects [TP53 gene therapy with chemotherapeutic drugs such as cisplatin synergized to enhance clinical efficacy [TP53 gene therapy in ovarian cancer was closed after the first interim analysis because adequate therapeutic benefit was not achieved [One alternative approach is the idea of restoring WT p53 function. The concept of restoring WT p53 activity is strongly supported by n humans –105. Var effects . Moreoveefficacy . UnfortuOther approaches such as targeting cellular proteins responsible for stabilizing mutant p53 are another route that may bring success. A recent development involves inhibiting the heat-shock protein HSP90, which chaperones many mutant p53 proteins ,107 and TP53 mutations.In summary, the information gained from studying the mutant p53 transcriptome and interactome described in this review has solidified the foundation for the development of strategies that can one day be used to treat the large number of cancer patients that harbor


Fourth, there was a significant gliosis mainly affecting the lower cortex of both disorders. The data suggest that the pattern of cortical degeneration is different in sCJD and vCJD which may reflect differences in aetiology and the subsequent spread of prion pathology within the brain.The laminar distributions of the pathological changes in the cerebral cortex were compared in the prion diseases sporadic Creutzfeldt-Jakob disease (sCJD) and variant CJD (vCJD). First, in some cortical regions, the vacuolation (“spongiform change”) was more generally distributed across the cortex in sCJD. Second, there was greater neuronal loss in the upper cortex in vCJD and in the lower cortex in sCJD. Third, the “diffuse” and “florid” prion protein (PrP PrP) gene [sc) is acquired from growth hormone, dura mater grafts, or corneal transplantation [PrP gene [Four subtypes of the prion disease Creutzfeldt-Jakob disease (CJD) have been described to date including familial CJD (fCJD), linked to germline mutations of the prion protein (rP) gene , and iatantation . By contPrP gene , 4. In aPrP gene . VariantPrP gene . VariantPrP gene .sc in the form of discrete deposits or plaques [sc deposit in CJD. Hence, “florid” deposits are composed of a central “core” surrounded by a rim of small vacuoles and are especially characteristic of vCJD. Broad bundles of amyloid are present within the core, and the deposits resemble more closely the “classic” deposits typical of Alzheimer's disease (AD) [sc occurs in the form of “synaptic-type” deposits, and there are relatively few florid deposits or kuru plaques [Neuropathologically, CJD is characterized by the presence in the brain of vacuolation “spongiform change”, neuronal loss, a reactive astrocytosis, and the deposition of PrP plaques , 7. Therase (AD)  rather tase (AD) . In addiase (AD)  which arase (AD) . By cont plaques , 13. sc spreads into the brain in CJD, for example, direct neural transmission from the site of infection, replication of PrPsc in the spleen followed by neural entry through the spinal cord [Differences in pathology between subtypes of CJD could reflect variations in aetiology and the subsequent spread of prion pathology within the brain. Various hypotheses have been proposed to explain how PrPnal cord , infectinal cord , 16, andnal cord . In neurnal cord –20. Neocnal cord . Hence, PrP gene or family history of prion disease, and there was no evidence of the known types of iatrogenic aetiology. In the vCJD cases, the pattern of PrPsc deposition typical of these cases was observed with florid-type PrPsc deposits in the cerebral cortex, cerebellum, basal ganglia, thalamus, and brain stem [PrP gene and the presence of Type 1 or Type 2 isoforms of PrPsc [sc (M/M1). All vCJD cases studied were methionine/methionine (M/M) homozygotes at codon 129. In addition, the PrPsc characteristic of vCJD had a uniform glycotype (PrPsc Type 4) [Eleven cases each of sCJD and vCJD  .μm sections were stained with haematoxylin and eosin (H/E), cresyl violet, or immunostained against PrP using the monoclonal antibody 12F10 (dilution 1 : 250) which binds to a region of human PrP downstream of the neurotoxic domain adjacent to helix region 2: residues 142–160 [Blocks of the frontal cortex (B8) at the level of the genu of the corpus callosum, parietal cortex (B7) at the level of the splenium of the corpus callosum, occipital cortex including the calcarine sulcus, inferior temporal gyrus (B22), and parahippocampal gyrus (B28) were taken from each case. Tissue was fixed in 10% phosphate-buffered formal saline and embedded in paraffin wax. Sequential, coronal 7 sc deposits across the cortical lamainae was studied using methods similar to those of Duyckaerts et al. [sc deposits in sCJD, all pathological changes were counted in 50 × 250 μm sample fields, the larger dimension of the field being located parallel with the surface of the pia mater. An eyepiece micrometer was used as the sample field and was moved down each traverse one step at a time from the pia mater to white matter. Histological features of the section were used to correctly position the field. Counts from the five traverses were added together to study the distribution of the pathology within each cortical region. Because of the diffuse nature of the synaptic deposits in sCJD, these lesions were quantified by “lattice sampling”, that is, by counting the number of times the intersections of the grid lines encountered a PrPsc deposit [The distribution of the vacuolation, surviving neurons, glial cell nuclei, and PrPs et al. . Five tr deposit . r”) and a significant reduction in the residual sums of squares compared with the preceding curves [χ2) contingency table tests.No attempt was made to locate precisely the boundaries between individual cortical laminae. First, the degree of pathological change and cell losses in the CJD cases made laminar identification difficult. Second, identification is especially difficult in the frontal cortex because it exhibits a heterotypical structure, that is, six laminae cannot be clearly identified even when the cortex is fully developed. Instead, variations in lesion density with distance below the pia mater were analysed using a curve-fitting procedure  , 29. Heng curves . The dissc and reveals the strongly immunolabeled florid PrPsc deposits which are composed of a condensed core of PrPsc and the more lightly immunolabeled and irregularly shaped diffuse deposits.sc deposits in the CJD cases studied are shown in sc deposits was fitted by a first-order polynomial  suggesting a linear decline in abundance with distance below the pia mater. By contrast, in sCJD , the distribution of the synaptic PrPsc deposits was fitted by a second-order (quadratic) polynomial  suggesting a peak of abundance in the lower cortical laminae. Typical examples of the laminar distribution of the PrPsc deposits in sCJD and vCJD. The diffuse and florid PrPsc deposits were more frequently distributed in the upper cortex in vCJD while the synaptic deposits were predominantly distributed in the lower cortex in sCJD. There were no essential differences in the laminar distribution of histological features in allocortical areas, for example, PHG and isocortical areas in either sCJD or vCJD.A summary of the laminar distributions of the pathological changes in all brain areas is shown in There were similarities and differences in the laminar distributions of the histological and pathological changes in sCJD and vCJD. First, there were similarities in the distribution of the vacuoles in the two disorders, but in sCJD, there were more brain regions in which the vacuoles were distributed across all laminae while in vCJD, there were more regions in which the vacuolation spared the superficial laminae and the region immediately adjacent to the white matter. Hence, the vacuolation may be more extensively distributed across the laminae in more regions in sCJD than in vCJD.Second, there was a marked difference in the distribution of the surviving neurons in sCJD and vCJD. In normal control brain, cortical neurons are often bimodally distributed with peaks of density in the upper and lower cortex, the two peaks being asymmetric and the upper density peak being significantly larger than the lower density peak –20. In sχ2 contingency tables): (a) All categories including totals for the bimodal distributions: Vacuoles χ2 = 11.87 ; Neurons χ2 = 13.27 , Glial cells χ2 = 4.43 (P > .05); Diffuse PrPsc versus Synaptic PrPscχ2 = 15.72 , Florid PrPsc versus Synaptic PrPscχ2 = 17.94 ; (b) comparison of unimodal distributions only: Vacuoles χ2 = 0.48 ; Neurons χ2 = 3.12 , Glial cells χ2 = 3.23 ; Diffuse PrPsc versus Synaptic PrPscχ2 = 12.55 , Florid PrPsc versus Synaptic PrPscχ2 = 13.37 ; (c) comparison of bimodal distributions only: Vacuoles χ2 = 4.21 , Neurons χ2 = 7.56 , Glia cell nuclei χ2 = 0.67 , Nonflorid PrPsc  χ2 versus Synaptic PrPscχ2 = 2.76 , Florid PrPsc versus Synaptic PrPscχ2 = 2.76 .Comparisons between sCJD and vCJD (sc deposition, this is unlikely to be the case in vCJD where neuronal losses appear to be greater in the upper cortical laminae. It is possible that in sCJD, the glial cell reaction directly reflects pathological changes in the lower laminae but in vCJD reflects greater degeneration of the afferent and efferent subcortical pathways which have their cells of origin or axon terminals in the lower cortex.Third, there are similarities in the distribution of glial cell nuclei in sCJD and vCJD which showed a marked preference for the lower cortical laminae, a distribution also reported in previous studies . Althougsc deposits were observed in the two disorders. In vCJD, the diffuse and florid PrPsc deposits showed a marked preference for the upper cortical laminae while the synaptic deposits of sCJD were more frequently distributed in the lower cortex. These distributions could be related to the degeneration of different anatomical pathways in the neocortex. Hence, the distribution of the pathological changes in vCJD suggests degeneration of the feed-forward corticocortical projections while degeneration of the feed-back corticocortical pathways and/or the feedback pathways may be present in sCJD [Fourth, significant differences in the laminar distribution of PrP in sCJD , 31. PrP gene [in situ, and the distribution of the pathology will reflect the site of origin and the pathways by which prion pathology spreads from these sites. Hence, if there are multiple sites of origin of the pathology, there may be a less marked topographic pattern to the pathology in sCJD than in vCJD.Differences in the distribution of the pathology in sCJD and vCJD could reflect the differences in the aetiology, and subsequent spread of prion pathology. Hence in vCJD, prions are believed to enter the nervous system by absorption through the gut following consumption of infected food, replication in the spleen, and neural entry via the spinal cord  whereas PrP gene , 4. The PrP gene . SubsequPrP gene . By contThe data are consistent with the hypothesis that there are different patterns of cortical degeneration in sCJD and vCJD. All laminae of the neocortex may be affected in both sCJD and vCJD, but, there is a greater development of the pathology in the lower laminae in sCJD and in the upper laminae in vCJD. Differences in cortical degeneration may be directly related to the different aetiologies of the two disorders and differences in the pattern of spread of the pathology through the brain.