Early Region Research Articles

Viral tumor antigen expression is no longer required in radiation-resistant subpopulation of JCV induced mouse medulloblastoma cells.

The human neurotropic polyomavirus JC, JC virus (JCV), infects the majority of human population during early childhood and establishes a latent/persistent infection for the rest of the life. JCV is the etiologic agent of the fatal demyelinating disease of the central nervous system, progressive multifocal leukoencephalopathy (PML) that is seen primarily in immunocompromised individuals. In addition to the PML, JCV has also been shown to transform cells in culture systems and cause a variety of tumors in experimental animals. Moreover, JCV genomic DNA and tumor antigen expression have been shown in a variety of human tumors with CNS origin. Similar to all polyomaviruses, JCV encodes for several tumor antigens from a single transcript of early coding region via alternative splicing. There is little known regarding the characteristics of JCV induced tumors and impact of DNA damage induced by radiation on viral tumor antigen expression and growth of these cells. Here we analyzed the possible impact of ionizing radiation on transformed phenotype and tumor antigen expression by utilizing a mouse medulloblastoma cell line (BSB8) obtained from a mouse transgenic for JCV tumor antigens. Our results suggest that a small subset of BSB8 cells survives and shows radiation resistance. Further analysis of the transformed phenotype of radiation resistant BSB8 cells (BSB8-RR) have revealed that they are capable of forming significantly higher numbers and sizes of colonies under anchorage dependent and independent conditions with reduced viral tumor antigen expression. Moreover, BSB8-RR cells show an increased rate of double-strand DNA break repair by homologous recombination (HR). More interestingly, knockout studies of JCV tumor antigens by utilizing CRISPR/Cas9 gene editing reveal that unlike parental BSB8 cells, BSB8-RR cells are no longer required the expression of viral tumor antigens in order to maintain transformed phenotype.

Transformation-induced stress at telomeres is counteracted through changes in the telomeric proteome including SAMHD1.

Telomeres play crucial roles during tumorigenesis, inducing cellular senescence upon telomere shortening and extensive chromosome instability during telomere crisis. However, it has not been investigated if and how cellular transformation and oncogenic stress alter telomeric chromatin composition and function. Here, we transform human fibroblasts by consecutive transduction with vectors expressing hTERT, the SV40 early region, and activated H-RasV12. Pairwise comparisons of the telomeric proteome during different stages of transformation reveal up-regulation of proteins involved in chromatin remodeling, DNA repair, and replication at chromosome ends. Depletion of several of these proteins induces telomere fragility, indicating their roles in replication of telomeric DNA. Depletion of SAMHD1, which has reported roles in DNA resection and homology-directed repair, leads to telomere breakage events in cells deprived of the shelterin component TRF1. Thus, our analysis identifies factors, which accumulate at telomeres during cellular transformation to promote telomere replication and repair, resisting oncogene-borne telomere replication stress.

FV6. Intention attribution and neural processing of laughter in female and male adolescents with conduct disorder

Introduction Children and adolescents with aggressive behavior tendencies show difficulties in correctly determining the social intention of others, tending to interpret the behavior of others as being intentionally hostile. Previously, we have shown distinct neural correlates of attributing hostile intent with increasing aggression scores within a community sample of adolescents. The aim of the current study was to investigate these neural processes among adolescents with a diagnosis of conduct disorder (CD) compared to a healthy control group (CG), while considering gender by diagnosis interactions. Methods 47 adolescents with CD (M 14.2, SD 2.01) and 65 adolescents without any psychiatric diagnoses (M 15.1, SD 2.41) rated 1.5 s audiovisual laugh sequences in a 3T MRI scanner. 20 friendly, 20 tickling and 20 taunting sequences were rated as friendly or taunting on a four-point scale. Rating differences were calculated with a 2 (gender) × 2 (group: CD vs CG) × 3 (laughter type) mixed model ANOVA. Neural responses were calculated in SPM8 in a whole brain analysis of the BOLD response to each laughter type and the correlation with hostile ratings of laughter intention. Results Overall, CD rated laughter as more hostile than CG (F(2, 107) = 5.940, p . 05 ), yet a significant laughter type × group interaction (F(2, 107) = 5.944, p . 001 ) showed this to be driven by friendly laughter. In neural response, CD > CG showed increased activation in the right inferior parietal lobe (IPL; x = 57, y = −51, z = 42; k = 313; Tpeak = 4.97; pFWE females showed increased activation in medial prefrontal (mPFC), extending into dorsal anterior cingulate cortices (dACC) (x = −6, y = 54, z = 9; k = 317; Tpeak = 4.59; pFWE Conclusions The current findings show deviant processing of social signals among adolescents with conduct disorder. CD participants showed heightened activation in early auditory processing regions (MTG) as well as more hostile ratings of friendly laughter. These results partially replicate previous findings in a population-based sample, emphasizing the importance of early sensory processing during social intention attribution in aggressive participants. Males showed increased activation of the areas implicated in both the mentalizing and social pain networks, mPFC and dACC, during intention attribution of taunting laughter. This unexpected result speaks for heightened emotional processing in males in response to signals of social exclusion.

Effects of Delayed Cord Clamping on 4-Month Ferritin Levels, Brain Myelin Content, and Neurodevelopment: A Randomized Controlled Trial

ObjectiveTo evaluate whether placental transfusion influences brain myelination at 4 months of age.Study designA partially blinded, randomized controlled trial was conducted at a level III maternity hospital in the US. Seventy-three healthy term pregnant women and their singleton fetuses were randomized to either delayed umbilical cord clamping (DCC, >5 minutes) or immediate clamping (ICC, <20 seconds). At 4 months of age, blood was drawn for ferritin levels. Neurodevelopmental testing (Mullen Scales of Early Learning) was administered, and brain myelin content was measured with magnetic resonance imaging. Correlations between myelin content and ferritin levels and group-wise DCC vs ICC brain myelin content were completed.ResultsIn the DCC and ICC groups, clamping time was 172 ± 188 seconds vs 28 ± 76 seconds (P < .002), respectively; the 48-hour hematocrit was 57.6% vs 53.1% (P < .01). At 4 months, infants with DCC had significantly greater ferritin levels (96.4 vs 65.3 ng/dL, P = .03). There was a positive relationship between ferritin and myelin content. Infants randomized to the DCC group had greater myelin content in the internal capsule and other early maturing brain regions associated with motor, visual, and sensory processing/function. No differences were seen between groups in the Mullen testing.ConclusionAt 4 months, infants born at term receiving DCC had greater ferritin levels and increased brain myelin in areas important for early life functional development. Endowment of iron-rich red blood cells obtained through DCC may offer a longitudinal advantage for early white matter development.Trial registrationClinicalTrials.gov: NCT01620008.

Confident false memories for spatial location are mediated by V1

ABSTRACTPrior functional magnetic resonance imaging (fMRI) results suggest that true memories, but not false memories, activate early sensory cortex. It is thought that false memories, which reflect conscious processing, do not activate early sensory cortex because these regions are associated with nonconscious processing. We posited that false memories may activate the earliest visual cortical processing region (i.e., V1) when task conditions are manipulated to evoke conscious processing in this region. In an fMRI experiment, abstract shapes were presented to the left or right of fixation during encoding. During retrieval, old shapes were presented at fixation and participants characterized each shape as previously on the "left" or "right" followed by an "unsure"–"sure"–"very sure" confidence rating. False memories for spatial location (i.e., "right"/left or "left"/right trials with "sure" or "very sure" confidence ratings) were associated with activity in bilateral early visual regions, including V1. In a follow-up fMRI-guided transcranial magnetic stimulation (TMS) experiment that employed the same paradigm, we assessed whether V1 activity was necessary for false memory construction. Between the encoding phase and the retrieval phase of each run, TMS (1 Hz, 8 min) was used to target the location of false memory activity (identified in the fMRI experiment) in left V1, right V1, or the vertex (control site). Confident false memories for spatial location were significantly reduced following TMS to V1, as compared to vertex. The results of the present experiments provide convergent evidence that early sensory cortex can contribute to false memory construction under particular task conditions.

Developing incrementality in filler-gap dependency processing

Much work has demonstrated that children are able to use bottom-up linguistic cues to incrementally interpret sentences, but there is little understanding of the extent to which children’s comprehension mechanisms are guided by top-down linguistic information that can be learned from distributional regularities in the input. Using a visual world eye tracking experiment and a corpus analysis, the current study investigates whether 5- and 6-year-old children incrementally assign interpretations to temporarily ambiguous wh-questions like What was Emily eating the cake with __? In the visual world eye-tracking experiment, adults demonstrated evidence for active dependency formation at the earliest region (i.e., the verb region), while 6-year-old children demonstrated a spill-over effect of this bias in the subsequent NP region. No evidence for this bias was found in 5-year-olds, although the speed of arrival at the ultimately correct instrument interpretation appears to be modulated by the vocabulary size. These results suggest that adult-like active formation of filler-gap dependencies begins to emerge around age 6. The corpus analysis of filler-gap dependency structures in adult corpora and child corpora demonstrate that the distributional regularities in either corpora are equally in favor of early, incremental completion of filler-gap dependencies, suggesting that the distributional information in the input is either not relevant to this incremental bias, or that 5-year-old children are somehow unable to recruit this information in real-time comprehension. Taken together, these findings shed light on the origin of the incremental processing bias in filler-gap dependency processing, as well as on the role of language experience and cognitive constraints in the development of incremental sentence processing mechanisms.

Tactile-to-Visual Cross-Modal Transfer of Texture Categorisation Following Training: An fMRI Study.

We investigated the neural underpinnings of texture categorisation using exemplars that were previously learned either within modalities (visual training and visual test) or across modalities (tactile training and visual test). Previous models of learning suggest a decrease in activation in brain regions that are typically involved in cognitive control during task acquisition, but a concomitant increase in activation in brain regions associated with the representation of the acquired information. In our study, participants were required to learn to categorise fabrics of different textures as either natural or synthetic. Training occurred over several sessions, with each fabric presented either visually or through touch to a participant. Pre- and post-training tests, in which participants categorised visual images only of the fabrics, were conducted during a functional magnetic resonance imaging (fMRI) scan. Consistent with previous research on cognitive processes involved in task acquisition, we found that categorisation training was associated with a decrease in activation in brain regions associated with cognitive systems involved in learning, including the superior parietal cortex, dorsal anterior cingulate cortex (dACC), and the right dorsolateral prefrontal cortex (DLFC). Moreover, these decreases were independent of training modality. In contrast, we found greater activation to visual textures in a region within the left medial occipital cortex (MOC) following training. There was no overall evidence of an effect of training modality in the main analyses, with texture-specific regional changes associated with both within- (visual) and cross- (touch) modal training. However, further analyses suggested that, unlike categorisation performance following within-modal training, crossmodal training was associated with bilateral activation of the MOC. Our results support previous evidence for a multisensory representation of texture within early visual regions of the cortex and provide insight into how multisensory categories are formed in the brain.

Degradation of a Novel DNA Damage Response Protein, Tankyrase 1 Binding Protein 1, following Adenovirus Infection

ABSTRACTInfection by most DNA viruses activates a cellular DNA damage response (DDR), which may be to the detriment or advantage of the virus. In the case of adenoviruses, they neutralize antiviral effects of DDR activation by targeting a number of proteins for rapid proteasome-mediated degradation. We have now identified a novel DDR protein, tankyrase 1 binding protein 1 (TNKS1BP1) (also known as Tab182), which is degraded during infection by adenovirus serotype 5 and adenovirus serotype 12. In both cases, degradation requires the action of the early region 1B55K (E1B55K) and early region 4 open reading frame 6 (E4orf6) viral proteins and is mediated through the proteasome by the action of cullin-based cellular E3 ligases. The degradation of Tab182 appears to be serotype specific, as the protein remains relatively stable following infection with adenovirus serotypes 4, 7, 9, and 11. We have gone on to confirm that Tab182 is an integral component of the CNOT complex, which has transcriptional regulatory, deadenylation, and E3 ligase activities. The levels of at least 2 other members of the complex (CNOT3 and CNOT7) are also reduced during adenovirus infection, whereas the levels of CNOT4 and CNOT1 remain stable. The depletion of Tab182 with small interfering RNA (siRNA) enhances the expression of early region 1A proteins (E1As) to a limited extent during adenovirus infection, but the depletion of CNOT1 is particularly advantageous to the virus and results in a marked increase in the expression of adenovirus early proteins. In addition, the depletion of Tab182 and CNOT1 results in a limited increase in the viral DNA level during infection. We conclude that the cellular CNOT complex is a previously unidentified major target for adenoviruses during infection.IMPORTANCE Adenoviruses target a number of cellular proteins involved in the DNA damage response for rapid degradation. We have now shown that Tab182, which we have confirmed to be an integral component of the mammalian CNOT complex, is degraded following infection by adenovirus serotypes 5 and 12. This requires the viral E1B55K and E4orf6 proteins and is mediated by cullin-based E3 ligases and the proteasome. In addition to Tab182, the levels of other CNOT proteins are also reduced during adenovirus infection. Thus, CNOT3 and CNOT7, for example, are degraded, whereas CNOT4 and CNOT1 are not. The siRNA-mediated depletion of components of the complex enhances the expression of adenovirus early proteins and increases the concentration of viral DNA produced during infection. This study highlights a novel protein complex, CNOT, which is targeted for adenovirus-mediated protein degradation. To our knowledge, this is the first time that the CNOT complex has been identified as an adenoviral target.

Temozolomide Enhances Triple-Negative Breast Cancer Virotherapy In Vitro.

Triple-negative breast cancer (TNBC) is one of the most aggressive types of cancer, and treatment is limited to chemotherapy and radiation. Oncolytic virotherapy may be a promising approach to treat TNBC. However, oncolytic adenovirus (OAd)-based mono-therapeutic clinical trials have resulted in modest outcomes. The OAd potency could be increased by chemotherapy-induced autophagy, an intracellular degradation system that delivers cytoplasmic constituents to the lysosome. In this study, the ability of alkylating agent temozolomide (TMZ)-induced autophagy to increase OAd replication and oncolysis in TNBC cells was evaluated. Human TNBC MDA-MB-231 and HCC1937 cells and mouse 4T1 cells were infected with an OAd expressing the red fluorescent protein mCherry on the virus capsid (OAdmCherry) alone or in combination with TMZ. TNBC cells treated with OAdmCherry/TMZ displayed greater mCherry and adenovirus (Ad) early region 1A (E1A) expression and enhanced cancer-cell killing compared to OAdmCherry or TMZ alone. The combined therapy-mediated cell death was associated with virus replication and accumulation of the autophagy marker light chain 3 (LC3)-II. Overall, this study provides experimental evidence of TMZ’s ability to increase oncolytic virotherapy in both human and murine TNBC cells.

Mapping Frequency-Specific Tone Predictions in the Human Auditory Cortex at High Spatial Resolution.

Auditory inputs reaching our ears are often incomplete, but our brains nevertheless transform them into rich and complete perceptual phenomena such as meaningful conversations or pleasurable music. It has been hypothesized that our brains extract regularities in inputs, which enables us to predict the upcoming stimuli, leading to efficient sensory processing. However, it is unclear whether tone predictions are encoded with similar specificity as perceived signals. Here, we used high-field fMRI to investigate whether human auditory regions encode one of the most defining characteristics of auditory perception: the frequency of predicted tones. Two pairs of tone sequences were presented in ascending or descending directions, with the last tone omitted in half of the trials. Every pair of incomplete sequences contained identical sounds, but was associated with different expectations about the last tone (a high- or low-frequency target). This allowed us to disambiguate predictive signaling from sensory-driven processing. We recorded fMRI responses from eight female participants during passive listening to complete and incomplete sequences. Inspection of specificity and spatial patterns of responses revealed that target frequencies were encoded similarly during their presentations, as well as during omissions, suggesting frequency-specific encoding of predicted tones in the auditory cortex (AC). Importantly, frequency specificity of predictive signaling was observed already at the earliest levels of auditory cortical hierarchy: in the primary AC. Our findings provide evidence for content-specific predictive processing starting at the earliest cortical levels.SIGNIFICANCE STATEMENT Given the abundance of sensory information around us in any given moment, it has been proposed that our brain uses contextual information to prioritize and form predictions about incoming signals. However, there remains a surprising lack of understanding of the specificity and content of such prediction signaling; for example, whether a predicted tone is encoded with similar specificity as a perceived tone. Here, we show that early auditory regions encode the frequency of a tone that is predicted yet omitted. Our findings contribute to the understanding of how expectations shape sound processing in the human auditory cortex and provide further insights into how contextual information influences computations in neuronal circuits.

The effect of implementation of an early detection team: A nationwide register-based study of characteristics and help-seeking behavior in first-episode schizophrenia in Denmark

BackgroundIn an effort to make people with signs of psychosis seek help as early as possible, Region Zealand launched in 2012 an early detection team project as the first and only in Denmark. The project consisted of a combination of easy access and an information campaign targeting the public. This nation-wide study examined characteristics and help-seeking behavior of patients with first-episode schizophrenia (FES) in the early detection region in comparison with other Danish regions. MethodData from the Danish National Schizophrenia register on all Danish patients diagnosed with first-episode schizophrenia during 2012 to 2015 were linked to demographic and health care data drawn from official national registers. Binary logistic regression analyses examined the difference between the early detection region and other regions controlling for demographic characteristics and utilization of mental health care services and contacts to general practitioner (GP). ResultsPatients in the early detection region were younger (OR = 0.51; CI: 0.42–0.62; p < 0.000) than in regions without early detection teams. Furthermore, they were more likely to be of Danish origin, and less likely to have contact with mental health services and GPs prior to FES. ConclusionThe study suggests that implementing an early detection team in combination with an information campaign contributed to detecting patients with first-episode schizophrenia earlier than in regions without the early detection team. The study gives an indication of different pathways among patients in the early detection region.

Protein E6 in High Risk Human Papillomaviruses

Human Papillomavirus (HPV) refers to a group of more than one hundred HPV high risk or low risk strains associated with a variety of conditions. Over fifty strains lead to genital infections, while others influence the development of various cancers, such as cervical, skin, etc. The combination of HPV E6/E7, hTERT, and Ki67 mRNA expression levels complement each other to diagnose lesions of the aforementioned cancers, specifically dealing with the cervix. All HPV strains share a common structure: circular, double stranded DNA encoding for eight open reading frames (ORF) surrounded by a non‐enveloped capsid. ORFs function to produce the E1‐E7 proteins needed for HPV replication (early region), L1‐L2 proteins that make up the capsid (late region), and replicate and transcribe viral DNA with the help of cis components (long control region). The E6, in particular, is an oncoprotein composed of polypeptides made up of 150 amino acids and a Cys‐X‐X‐Cys motif that allow for the formation of two zinc fingers. E6 and E7 recognize a number of host proteins by hijacking their cellular domain‐motif interaction networks. In the HPV‐16 and HPV‐18 strains, the E6 protein binds its amino and carboxyl terminal regions to a leucine‐rich sequence (LxxLL) within the cellular ubiquitin ligase, E6AP. This heterodimer degrades p53, a proapoptotic tumor suppressor, although the exact mode of assembly is unknown. Complete degradation potentially leads to oncogenesis in high‐risk HPV strains, as these transformed cells have lost their ability to arrest at the G1 checkpoint within their cell cycle and perform apoptosis. Aside from the E6‐E6AP heterodimer binding directly to p53, the E6 protein has other oncogenic functions, allowing it to bind to four general classes of other proteins, providing indirect pathways to degrade p53. Binding options include proteins involved in cell polarity and motility, tumor suppressors and inducers of apoptosis, transcriptional coactivators and DNA replication/repair factors. Regarding transcription coactivators, degradation of p53 transcriptional activity is possible, as seen by the targeting of p53 coactivator p300/CBP or yADA3. Given the mechanism of E6‐E6AP complex hindering p53's from carrying out normal function, E6 is an attractive target for therapies against cervical cancers. The Olathe North High School MSOE Center for BioMolecular Modeling SMART Team used 3‐D modeling and printing technology to examine structure‐function relationships of E6.Support or Funding InformationSMART team is a joint venture between the Milwaukee School of Engineering and the Olathe School District's 21st century program Medical Professions Academy.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Quantifying the anthropocene loss of bioindicators for an early industrial region: an equitable baseline for biodiversity restoration

Directly observed biodiversity data have a limited temporal span of c. 100–150 years. Consequently, for a region such as temperate Western Europe, our knowledge of species distributions is restricted to a period impacted by the process of massive industrialisation. There is a danger of shifted baselines in terms of conservation policy and targets. Here we present a novel source of high resolution archaeobotanical information for lichen epiphyte bioindicators; these data can reconstruct species distributions for the pre-industrial European landscape. We compare these historic records to a species’ post-industrial distribution and environmental response, quantifying the spatial trend and causes of biodiversity loss. The results indicate regional extinction rates of c. 76% in response to habitat loss and industrial pollution. We propose pre-industrial baselines that would better represent biodiversity restoration for temperate regions (net gain), and which would be equitable with advocacy for species and habitat protection in the present-day tropics (no net loss).

CTCF Binding Sites in the Herpes Simplex Virus 1 Genome Display Site-Specific CTCF Occupation, Protein Recruitment, and Insulator Function.

There are seven conserved CTCF binding domains in the herpes simplex virus 1 (HSV-1) genome. These binding sites individually flank the latency-associated transcript (LAT) and the immediate early (IE) gene regions, suggesting that CTCF insulators differentially control transcriptional domains in HSV-1 latency. In this work, we show that two CTCF binding motifs in HSV-1 display enhancer blocking in a cell-type-specific manner. We found that CTCF binding to the latent HSV-1 genome was LAT dependent and that the quantity of bound CTCF was site specific. Following reactivation, CTCF eviction was dynamic, suggesting that each CTCF site was independently regulated. We explored whether CTCF sites recruit the polycomb-repressive complex 2 (PRC2) to establish repressive domains through a CTCF-Suz12 interaction and found that Suz12 colocalized to the CTCF insulators flanking the ICP0 and ICP4 regions and, conversely, was removed at early times postreactivation. Collectively, these data support the idea that CTCF sites in HSV-1 are independently regulated and may contribute to lytic-latent HSV-1 control in a site-specific manner.IMPORTANCE The role of chromatin insulators in DNA viruses is an area of interest. It has been shown in several beta- and gammaherpesviruses that insulators likely control the lytic transcriptional profile through protein recruitment and through the formation of three-dimensional (3D) chromatin loops. The ability of insulators to regulate alphaherpesviruses has been understudied to date. The alphaherpesvirus HSV-1 has seven conserved insulator binding motifs that flank regions of the genome known to contribute to the establishment of latency. Our work presented here contributes to the understanding of how insulators control transcription of HSV-1.

Co-infection with human polyomavirus BK enhances gene expression and replication of human adenovirus

Immunocompromised patients are susceptible to multiple viral infections. Relevant interactions between co-infecting viruses might result from viral regulatory genes which trans-activate or repress the expression of host cell genes as well as the genes of any co-infecting virus. The aim of the current study was to show that the replication of human adenovirus 5 is enhanced by co-infection with BK polyomavirus and is associated with increased expression of proteins including early region 4 open reading frame 1 and both the large tumor antigen and small tumor antigen. Clinical samples of whole blood and urine from 156 hematopoietic stem cell transplant recipients were tested. We also inoculated adenocarcinomic human alveolar basal epithelial cells with both human adenovirus 5 and BK polyomavirus to evaluate if co-infection of viruses affected their replication. Data showed that adenovirus load was significantly higher in the plasma (mean 7.5 x 103 ± 8.5 x 102 copies/ml) and urine (mean 1.9 x 103 ± 8.0 x 102 copies/ml) of samples from patients with co-infections, in comparison to samples from patients with isolated adenovirus infection. In vitro co-infection led to an increased (8.6 times) expression of the adenovirus early region 4 open reading frame gene 48 hours post-inoculation. The expression of the early region 4 open reading frame gene positively correlated with the expression of BK polyomavirus large tumor antigen (r = 0.90, p < 0.0001) and small tumor antigen (r = 0.83, p < 0.001) genes. The enhanced expression of the early region 4 open reading frame gene due to co-infection with BK polyomavirus was associated with enhanced adenovirus, but not BK polyomavirus, replication. The current study provides evidence that co-infection of adenovirus and BK polyomavirus contributes to enhanced adenovirus replication. Data obtained from this study may have significant importance in the clinical setting.

HPV-Induced Field Cancerisation: Transformation of Adult Tissue Stem Cell Into Cancer Stem Cell.

Field cancerisation was originally described as a basis for multiple head and neck squamous cell carcinoma (HNSCC) and is a pre-malignant phenomenon that is frequently attributable to oncogenic human papillomavirus (HPV) infection. Our work on β-HPV-induced cutaneous squamous cell carcinomas identified a novel Lrig1+ hair follicle junctional zone keratinocyte stem cell population as the basis for field cancerisation. Herein, we describe the ability for HPV to infect adult tissue stem cells in order to establish persistent infection and induce their proliferation and displacement resulting in field cancerisation. By review of the HPV literature, we reveal how this mechanism is conserved as the basis of field cancerisation across many tissues. New insights have identified the capacity for HPV early region genes to dysregulate adult tissue stem cell self-renewal pathways ensuring that the expanded population preserve its stem cell characteristics beyond the stem cell niche. HPV-infected cells acquire additional transforming mutations that can give rise to intraepithelial neoplasia (IEN), from environmental factors such as sunlight or tobacco induced mutations in skin and oral cavity, respectively. With establishment of IEN, HPV viral replication is sacrificed with loss of the episome, and the tissue is predisposed to multiple cancer stem cell-driven carcinomas.

Connectivity of the ventral visual cortex is necessary for object recognition in patients.

The functional profiles of regions in the ventral occipital-temporal cortex (VTC), a critical region for object visual recognition, are associated with the VTC connectivity patterns to nonvisual regions relevant to the corresponding object domain. However, whether and how whole-brain connections affect recognition behavior remains untested. We directly examined the necessity of VTC connectivity in object recognition behavior by testing 82 patients whose lesion spared relevant VTC regions but affected various white matter (WM) tracts and other regions. In these patients, we extracted the whole-brain anatomical connections of two VTC domain-selective (large manmade objects and animals) clusters with probabilistic tractography, and examined whether such connectivity pattern can predict recognition performance of the corresponding domains with support vector regression (SVR) analysis. We found that the whole-brain anatomical connectivity of large manmade object-specific cluster successfully predicted patients' large object recognition performance but not animal recognition or control tasks, even after we excluded connections with early visual regions. The contributing connections to large object recognition included tracts between VTC-large object cluster and distributed regions both within and beyond the visual cortex (e.g., putamen, superior, and middle temporal gyrus). These results provide causal evidence that the VTC whole-brain anatomical connectivity is necessary for at least certain domains of object recognition behavior.

Ptolemaic military operations were a dominant factor in the spread of Egyptian cults across the early Hellenistic Aegean Sea.

Early in the Ptolemaic era, Egyptian cults, particularly those of Isis and Sarapis, spread successfully to ports across the ancient Aegean Sea. Leading researchers in the field claim that the spread of these cults was influenced by multiple factors, ones that were mainly economic or political in character. However, the question of which factors had more weight or impact than others in the process of the early spread of Egyptian cults has not yet been answered in academic discussion. This could be related to the fact that the issue of the spread of religious innovations in the ancient Mediterranean has been addressed mainly by established historiographical methods such as the collection and critical analysis of archaeological and literary sources. Hypotheses and conclusions derived from these methods are, however, often unable to reflect the complexity of historical processes. A possible solution can be found in supplementing this established methodological apparatus by formalized methods, e.g. the coding of relevant datasets, statistics, geospatial modeling, and network analysis. To be able to compare the possible impacts of different factors on the spread of Egyptian cults in the Aegean Sea region, we 1) constructed a model of the ancient maritime transportation network as a platform for quantitative analysis, 2) transformed selected factors of possible influence into georeferenced parameters of the network, and 3) defined a mathematical model that allowed us to determine which parameters of the network explain the spatial dissemination of archaeological evidence connected to Egyptian cults. The results suggest that the most significant correlation is between the placement of Ptolemaic garrisons and the distribution of Egyptian temples and artefacts in the early Hellenistic Aegean Sea region. The interpretation would be that Egyptian military forces potentially played a significant role in the spread of Egyptian cults.

Novel Human Polyomavirus Noncoding Control Regions Differ in Bidirectional Gene Expression according to Host Cell, Large T-Antigen Expression, and Clinically Occurring Rearrangements.

Human polyomavirus (HPyV) DNA genomes contain three regions denoted the early viral gene region (EVGR), encoding the regulatory T-antigens and one microRNA, the late viral gene region (LVGR), encoding the structural Vp capsid proteins, and the noncoding control region (NCCR). The NCCR harbors the origin of viral genome replication and bidirectional promoter/enhancer functions governing EVGR and LVGR expression on opposite DNA strands. Despite principal similarities, HPyV NCCRs differ in length, sequence, and architecture. To functionally compare HPyV NCCRs, sequences from human isolates were inserted into a bidirectional reporter vector using dsRed2 for EVGR expression and green fluorescent protein (GFP) for LVGR expression. Transfecting HPyV NCCR reporter vectors into human embryonic kidney 293 (HEK293) cells and flow cytometry normalized to archetype BKPyV NCCR revealed a hierarchy of EVGR expression levels with MCPyV, HPyV12, and STLPyV NCCRs conferring stronger levels and HPyV6, HPyV9, and HPyV10 NCCRs weaker levels, while LVGR expression was less variable and showed comparable activity levels. Transfection of HEK293T cells expressing simian virus 40 (SV40) large T antigen (LTag) increased EVGR expression for most HPyV NCCRs, which correlated with the number of LTag-binding sites (Spearman's r, 0.625; P < 0.05) and decreased following SV40 LTag small interfering RNA (siRNA) knockdown. LTag-dependent activation was specifically confirmed for two different MCPyV NCCRs in 293MCT cells expressing the cognate MCPyV LTag. HPyV NCCR expression in different cell lines derived from skin (A375), cervix (HeLaNT), lung (A549), brain (Hs683), and colon (SW480) demonstrated that host cell properties significantly modulate the baseline HPyV NCCR activity, which partly synergized with SV40 LTag expression. Clinically occurring NCCR sequence rearrangements of HPyV7 PITT-1 and -2 and HPyV9 UF1 were found to increase EVGR expression compared to the respective HPyV archetype, but this was partly host cell type specific.IMPORTANCE HPyV NCCRs integrate essential viral functions with respect to host cell specificity, persistence, viral replication, and disease. Here, we show that HPyV NCCRs not only differ in sequence length, number, and position of LTag- and common transcription factor-binding sites but also confer differences in bidirectional viral gene expression. Importantly, EVGR reporter expression was significantly modulated by LTag expression and by host cell properties. Clinical sequence variants of HPyV7 and HPyV9 NCCRs containing deletions and insertions were associated with increased EVGR expression, similar to BKPyV and JCPyV rearrangements, emphasizing that HPyV NCCR sequences are major determinants not only of host cell tropism but also of pathogenicity. These results will help to define secondary HPyV cell tropism beyond HPyV surface receptors, to identify key viral and host factors shaping the viral life cycle, and to develop preclinical models of HPyV persistence and replication and suitable antiviral targets.

Rates of Mutation and Recombination in Siphoviridae Phage Genome Evolution over Three Decades.

The evolution of asexual organisms is driven not only by the inheritance of genetic modification but also by the acquisition of foreign DNA. The contribution of vertical and horizontal processes to genome evolution depends on their rates per year and is quantified by the ratio of recombination to mutation. These rates have been estimated for bacteria; however, no estimates have been reported for phages. Here, we delineate the contribution of mutation and recombination to dsDNA phage genome evolution. We analyzed 34 isolates of the 936 group of Siphoviridae phages using a Lactococcus lactis strain from a single dairy over 29 years. We estimate a constant substitution rate of 1.9 × 10−4 substitutions per site per year due to mutation that is within the range of estimates for eukaryotic RNA and DNA viruses. The reconstruction of recombination events reveals a constant rate of five recombination events per year and 4.5 × 10−3 nucleotide alterations due to recombination per site per year. Thus, the recombination rate exceeds the substitution rate, resulting in a relative effect of recombination to mutation (r/m) of ∼24 that is homogenous over time. Especially in the early transcriptional region, we detect frequent gene loss and regain due to recombination with phages of the 936 group, demonstrating the role of the 936 group pangenome as a reservoir of genetic variation. The observed substitution rate homogeneity conforms to the neutral theory of evolution; hence, the neutral theory can be applied to phage genome evolution and also to genetic variation brought about by recombination.