bluuebunny/crossref_metadata_2025 huggingface dataset:
Neuroendocrine and behavioral measures of stress-reactivity in male goal-tracker and sign-tracker rats
Authors: Lopez Sofia A. , Mubarak Eman , Yang Charlotte , Parsegian Aram , Klumpner Marin , Campus Paolo , Flagel Shelly B.
Published: September 2020

Environmental cues attain the ability to guide behavior via learned associations. As predictors, cues can elicit adaptive behavior and lead to valuable resources (e.g., food). For some individuals, however, cues are transformed into incentive stimuli and elicit motivational states that can be maladaptive. The goal-tracker/sign-tracker animal model captures individual differences in cue-motivated behaviors, with reward-associated cues serving as predictors of reward for both phenotypes but becoming incentive stimuli to a greater degree for sign-trackers. While these distinct phenotypes are characterized based on Pavlovian conditioned approach behavior, they exhibit differences on a number of behaviors relevant to psychopathology. To further characterize the neurobehavioral endophenotype associated with individual differences in cue-reward learning, neuroendocrine and behavioral profiles associated with stress and anxiety were investigated in male goal-tracker, sign-tracker, and intermediate responder rats. It was revealed that baseline corticosterone increases with Pavlovian learning, but to the same degree, regardless of phenotype. No significant differences in behavior were observed between goaltrackers and sign-trackers during an elevated plus maze or open field test, nor were there differences in corticosterone response to the open field test or physiological restraint. Upon examination of central markers associated with stress-reactivity, we found that sign-trackers have greater glucocorticoid receptor mRNA expression in the ventral hippocampus, with no phenotypic differences in the dorsal hippocampus or prelimbic cortex. These findings demonstrate that goal-trackers and sign-trackers do not differ on stress- and anxiety-related behaviors, and suggest that differences in neuroendocrine measures between these phenotypes can be attributed to distinct cue-reward learning styles.

Significance Statement

While the goal-tracker/ sign-tracker animal model derives from individual differences in Pavlovian conditioned approach behavior, other traits, including some of relevance to addiction and post-traumatic stress disorder, have been shown to co-exist with the propensity to sign-track. The extent to which this model encompasses differences in aversive arousal and associated neuroendocrine measures, however, remains largely unexplored. Here we show that behavioral and corticosterone response to stress-related paradigms do not differ between goal-trackers and sign-trackers. However, glucocorticoid receptor expression in the ventral hippocampus does differ between phenotypes, suggesting that this central marker that is typically associated with stress-responsivity, may, in fact, play an important role in appetitive motivation.
Identification of genetic factors controlling phosphorus utilization efficiency in wheat by genome-wide association study with principal component analysis
Authors: Safdar Luqman Bin , Umer Muhammad Jawad , Almas Fakhrah , Uddin Siraj , Safdar Qurra-tul-Ain , Blighe Kevin , Quraishi Umar Masood
Published: September 2020

Despite the economic importance of P utilization efficiency, information on genetic factors underlying this trait remains elusive. To address that, we performed a genome-wide association study in a spring wheat diversity panel ranging from landraces to elite varieties. We evaluated the phenotype variation for P utilization efficiency in controlled conditions and genotype variation using wheat 90K SNP array. Phenotype variables were transformed into a smaller set of uncorrelated principal components that captured the most important variation data. We identified two significant loci associated with both P utilization efficiency and the 1st principal component on chromosomes 3A and 4A:qPE1-3AandqPE2-4A. Annotation of genes at these loci revealed 53 wheat genes, among which 6 were identified in significantly enriched pathways. The expression pattern of these 6 genes indicated thatTraesCS4A02G481800, involved in pyruvate metabolism and TCA cycle, had a significantly higher expression in the P efficient variety under limited P conditions. Further characterization of these loci and candidate genes can help stimulate P utilization efficiency in wheat.

KEY MESSAGE

We report two new loci for P utilization efficiency on chromosomes 3A and 4A of wheat. The prioritized candidate genes at these loci can be investigated by molecular biology techniques to improve P efficiency in wheat and grass relatives.
Electrophysiological Indicators of Sleep-associated Memory Consolidation in 5- to 6-Year-Old Children
Authors: Joechner Ann-Kathrin , Wehmeier Sarah , Werkle-Bergner Markus
Published: September 2020

In adults, the synchronised interplay of sleep spindles (SP) and slow oscillations (SO) supports memory consolidation. Given tremendous developmental changes in SP and SO morphology, it remains elusive whether across childhood the same mechanisms as identified in adults are functional. Based on topography and frequency, we characterise slow and fast SPs and their temporal coupling to SOs in 24 pre-school children. Further, we ask whether slow and fast SPs and their modulation during SOs are associated with behavioural indicators of declarative memory consolidation as suggested by the literature on adults. Employing an individually tailored approach, we reliably identify an inherent, development-specific fast centro-parietal SP type, nested in the adult-like slow SP frequency range, along with a dominant slow frontal SP type. Further, we provide evidence that the modulation of fast centro-parietal SPs during SOs is already present in pre-school children. However, the temporal coordination between fast centro-parietal SPs and SOs is weaker and less precise than expected from research on adults. While we do not find evidence for a critical contribution of SP–SO coupling for memory consolidation, crucially, slow frontal and fast centro-parietal SPs are each differentially related to sleep-associated consolidation of items of varying quality. While a higher number of slow frontal SPs is associated with stronger maintenance of medium-quality memories, a higher number of fast centro-parietal SPs is linked to a greater gain of low-quality items. Our results demonstrate two functionally relevant inherent SP types in pre-school children although SP–SO coupling is not yet fully mature.
Stable 3D Head Direction Signals in the Primary Visual Cortex
Authors: Guitchounts Grigori , Lotter William , Dapello Joel , Cox David
Published: September 2020

Visual signals influence the brain’s computation of spatial position and orientation. Accordingly, the primary visual cortex (V1) is extensively interconnected with areas involved in computing head direction (HD) information. Predictive coding theories posit that higher cortical areas send sensory or motor predictions to lower areas, but whether this includes cognitive variables like the HD signal—and whether HD information is present in V1—is unknown. Here we show that V1 encodes the yaw, roll, and pitch of the head in freely behaving rats, either in the presence or absence of visual cues. HD tuning was modulated by lighting and movement state, but was stable on a population level for over a week. These results demonstrate the presence of a critical spatial orientation signal in a primary cortical sensory area and support predictive coding theories of brain function.
Integrated omics networks reveal the temporal signaling events of brassinosteroid response inArabidopsis
Authors: Clark Natalie M , Nolan Trevor M , Wang Ping , Song Gaoyuan , Montes Christian , Guo Hongqing , Sozzani Rosangela , Yin Yanhai , Walley Justin W
Published: September 2020

Brassinosteroids (BRs) are plant steroid hormones that are known to regulate cell division and stress response. We used a systems biology approach to integrate multi-omic datasets and unravel the molecular signaling events of BR response inArabidopsis. We profiled the levels of 32,549 transcripts, 9,035 protein groups, and 26,950 phosphorylation sites fromArabidopsisseedlings treated with brassinolide (BL, most active BR) for six different lengths of time. We then constructed a network inference pipeline called Spatiotemporal Clustering and Inference of Omics Networks (SC-ION) that was able to integrate these multi-omic data into one, cohesive regulatory network. Our network illustrates the signaling cascade of BR response, starting with kinase-substrate phosphorylation and ending with transcriptional regulation. We used our network predictions to identify putative, relevant phosphorylation sites on the TF BRI1-EMS-SUPPRESSOR 1 (BES1); the importance of which we experimentally validated. Additionally, we identified an uncharacterized TF, which we named BRONTOSAURUS (BRON), that regulates cell division. Further, we show thatbronmutant roots are hypersensitive to BL. Together, this work demonstrates the power of integrative network analysis applied to multiomic data and provides fundamental insights into the molecular signaling events occurring during BR response.
Soil characteristics constrain the response of bacterial and fungal communities and hydrocarbon degradation genes to phenanthrene soil contamination and phytoremediation with poplars
Authors: Correa-García Sara , Rheault Karelle , Tremblay Julien , Séguin Armand , Yergeau Etienne
Published: September 2020

Rhizodegradation is a promising cleanup technology where microorganisms degrade soil contaminants in the rhizosphere. A symbiotic relationship is expected to occur between plant roots and soil microorganisms in contaminated soils that enhance natural microbial degradation in soils. However, little is known about how this initial microbiota influences the rhizodegradation outcome in a context of different soil microbiotas. Recent studies have hinted that soil initial diversity has a determining effect on the outcome of contaminant degradation. To test this hypothesis, we planted (P) or not (NP) balsam poplars (Populus balsamifera) in two soils of contrasting diversity (agricultural and forest) that were contaminated or not with 50 mg kg-1of phenanthrene (PHE). The DNA from the rhizosphere of the P and the bulk soil of the NP pots was extracted and the bacterial genes encoding for the 16S rRNA, the PAH ring-hydroxylating dioxygenase alpha subunits (PAH-RHDα) of gram-positive (GP) and gram-negative (GN) bacteria, and the fungal ITS region were sequenced to characterize the microbial communities. and the abundance of the PAH-RHDα genes were also quantified by real-time quantitative PCR. Plant presence had a significant effect on PHE degradation only in the forest soil, whereas both NP and P agricultural soils degraded the same amount of PHE. Bacterial communities were principally affected by the soil type, and upon contamination the dominant PAH degrading community was similarly constrained by soil type. Our results highlight the crucial importance of soil microbial and physicochemical characteristics in the outcome of rhizoremediation.
Deletion of Mcpip1 in Mcpip1AlbKOmice recapitulates the phenotype of human primary biliary cholangitis
Authors: Kotlinowski Jerzy , Hutsch Tomasz , Czyzynska-Cichon Izabela , Wadowska Marta , Pydyn Natalia , Jasztal Agnieszka , Kij Agnieszka , Dobosz Ewelina , Lech Maciej , Miekus Katarzyna , Pośpiech Ewelina , Fu Mingui , Jura Jolanta , Koziel Joanna , Chlopicki Stefan
Published: September 2020

Background & Aims

Primary biliary cholangitis (PBC) is an autoimmune disease characterized by progressive destruction of the intrahepatic bile ducts. The immunopathology of PBC involves excessive inflammation; therefore, negative regulators of inflammatory response, such as Monocyte Chemoattractant Protein-1-Induced Protein-1 (MCPIP1,aliasRegnase1) may play important roles in the development of PBC. The aim of this work was to verify whether Mcpip1 expression protects against development of PBC.

Methods

Genetic deletion ofZc3h12awas used to characterize the role of Mcpip1 in the pathogenesis of PBC. 6-52-week-old Mcpip1fl/fland Mcpip1AlbKOmice were used for immunohistochemical, biochemical and molecular tests.

Results

We found that Mcpip1 deficiency in the liver recapitulates most of the features of human PBC, in contrast to mice with Mcpip1 deficiency in myeloid cells (Mcpip1LysMKOmice), which present with robust myeloid cell-driven systemic inflammation. In Mcpip1AlbKOlivers, intrahepatic bile ducts displayed proliferative changes with inflammatory infiltration, bile duct destruction, and fibrosis leading to cholestasis. In plasma, increased concentrations of IgG, IgM, and AMA autoantibodies (anti-PDC-E2) were detected. Interestingly, the phenotype of Mcpip1AlbKOmice was robust in 6-week-old and 52-week-old mice, but milder in 12-24-week-old mice, suggesting early prenatal origin of the phenotype and age-dependent progression of the disease. Hepatic transcriptome analysis of 6-week-old and 24-week-old Mcpip1AlbKOmice showed 812 and 8 differentially expressed genes (DEGs), respectively, compared with age-matched control mice, and revealed a distinct set of genes compared to those previously associated with development of PBC.

Conclusions

The phenotype of Mcpip1AlbKOmice recapitulates most of the features of human PBC, and demonstrates early prenatal origin and age-dependent progression of PBC. Therefore, Mcpip1AlbKOmice provide a unique model for the study of PBC.

Lay summary

Deletion of hepatic Mcpip1 in Mcpip1AlbKOmice leads to development of PBC that recapitulates phenotype of human patients. These animals, show early prenatal origin and age-dependent progression of the disease. Thus, Mcpip1AlbKOmice provide a unique model for studying PBC.
TRPV1 feed-forward sensitisation depends on COX2 upregulation in primary sensory neurons
Authors: Li Tianci , Wang Gaoge , Chin Hui Vivian Chin , Saad Daniel , de Sousa Valente Joao , Montanara Paolo La , Nagy Istvan
Published: September 2020

ABSRTRACT

Increased activity and excitability (sensitisation) of a series of molecules including the transient receptor potential ion channel, vanilloid subfamily, member 1 (TRPV1) in pain-sensing (nociceptive) primary sensory neurons are pivotal for developing pathological pain experiences in tissue injuries. TRPV1 sensitisation is induced and maintained by two major mechanisms; post-translational and transcriptional changes in TRPV1 induced by inflammatory mediators produced and accumulated in injured tissues, and TRPV1 activation-induced feed-forward signalling. The latter mechanism includes synthesis of TRPV1 agonists within minutes, and upregulation of various receptors functionally linked to TRPV1 within a few hours, in nociceptive primary sensory neurons. Here, we report that a novel mechanism, which contributes to TRPV1 activation-induced TRPV1-sensitisation within ~30 minutes in at least ~30% of TRPV1-expressing cultured murine primary sensory neurons, is mediated through upregulation in cyclooxygenase 2 (COX2) expression and increased synthesis of a series of COX2 products. These findings highlight the importance of feed-forward signalling in sensitisation, and the value of inhibiting COX2 activity to control pain, in nociceptive primary sensory neurons in tissue injuries.
RNA LEVER Mediates Long-Range Regulation of ε-globin by Keeping PRC2 in Check
Authors: Teo Wei Wen , Cao Xinang , Wu Chan-Shuo , Tan Hong Kee , Zhou Qiling , Yang Henry , Chai Li , Tenen Daniel G.
Published: September 2020

Polycomb Repressive Complex 2 (PRC2) is an epigenetic regulator required for gene silencing during embryonic development. Previous studies have reported that PRC2 interacts with RNA in a promiscuous manner, but the biological functions of such interaction are unknown. Here we present a seesaw mechanism for the regulation of ε-globin through inactivatingEZH2 by an upstream non-codingRNA (LEVER). We show that LEVER, a non-coding RNA identified by RNA immunoprecipitation sequencing (RIP-seq) of the PRC2 core subunit EZH2 and Nanopore sequencing, binds PRC2 and thereby prevents the accumulation of H3K27 methylation along the genomic region where LEVER RNA is transcribed. The open chromatin within the LEVER locus in turn competes for the chromatin interaction between the ε-globin promoter and the Locus Control Region (LCR), working as a negative regulatory element of ε-globin expression. Hence, LEVER RNA negatively regulates ε-globin by sequestering PRC2 from repressing the LEVER locus, which is a competitor of the ε-globin-LCR interaction.
rab-27acts in an intestinal secretory pathway to inhibit axon regeneration inC. elegans
Authors: Lin-Moore Alexander T. , Oyeyemi Motunrayo J. , Hammarlund Marc
Published: September 2020

Injured axons must regenerate to restore nervous system function, and regeneration is regulated in part by external factors from non-neuronal tissues. Many of these extrinsic factors act in the immediate cellular environment of the axon to promote or restrict regeneration, but the existence of long-distance signals regulating axon regeneration has not been clear. Here we show that the Rab GTPaserab-27inhibits regeneration of GABAergic motor neurons inC. elegansthrough activity in the intestine. Re-expression of RAB-27, but not the closely related RAB-3, in the intestine ofrab-27mutant animals is sufficient to rescue normal regeneration. Several additional components of an intestinal neuropeptide secretion pathway also inhibit axon regeneration, including NPDC1/cab-1, SNAP25/aex-4, and KPC3/aex-5. Together these data indicate that RAB-27-dependent neuropeptide secretion from the intestine inhibits axon regeneration, and point to distal tissues as potent extrinsic regulators of regeneration.
A myosin II-based nanomachine devised for the study of Ca2+-dependent mechanisms of muscle regulation
Authors: Pertici Irene , Bianchi Giulio , Bongini Lorenzo , Lombardi Vincenzo , Bianco Pasquale
Published: September 2020

The emergent properties of the array arrangement of the molecular motor myosin II in the sarcomere of the striated muscle, generation of steady force and shortening, can be studiedin vitrowith a synthetic nanomachine, made by an ensemble of eight HMM myosin fragments from rabbit psoas muscle carried on a piezoelectric nanopositioner and brought to interact with a properly oriented actin filament attached via gelsolin (a Ca2+-regulated actin binding protein) to a bead trapped by a Dual Laser Optical Tweezers. The application of the original version of the nanomachine to investigation of the Ca2+-dependent regulation mechanisms of the other sarcomeric (regulatory or cytoskeleton) proteins, adding them on at a time, was anyway prevented by the impossibility to preserve Ca2+as a free parameter. Here the nanomachine is implemented by assembling the bead-attached actin filament with the Ca2+-insensitive gelsolin fragment TL40. The performance of the nanomachine is determined either in the absence or in the presence of 0.1 mM Ca2+(the concentration required for BTA preparation with gelsolin). The nanomachine exhibits a maximum power output of 5.4 aW, independently of [Ca2+], opening the possibility for future studies of the Ca2+-dependent function/dysfunction of regulatory and cytoskeletal proteins.
Dispatched conformational dynamics couples transmembrane Na+flux to release of lipid-modified Hedgehog signal
Authors: Wang Qianqian , Asarnow Daniel E. , Ding Ke , Zhang Yunxiao , Ma Yong , Cheng Yifan , Beachy Philip A.
Published: September 2020

The DISP1 protein, related to the NPC1 and PTCH1 cholesterol transporters and to H+-driven transporters of the RND family, enables tissue patterning activity of the lipid-modified Hedgehog protein by releasing it from tightly-localized sites of embryonic expression. A 2.5 Å resolution cryo-EM structure of DISP1 revealed three Na+ions coordinated within a channel that traverses its transmembrane domain. This channel and Na+-binding are disrupted in DISP1-NNN (3.4 Å resolution), a variant with isosteric substitutions for three intramembrane aspartates that each coordinate and neutralize charge for one of the three Na+ions. DISP1-NNN and other variants that singly disrupt the Na+sites retain binding to the lipid-modified Hedgehog protein, but most fail to export Hedgehog. The Sonic hedgehog signal (ShhN) interacts with the DISP1 extracellular domains (2.7 Å complex structure), including an unusual extended DISP1 arm, at a location well above the membrane. Variability analysis reveals a dynamic series of DISP1 conformations, only a restricted subset of which appear to bind ShhN. The bound and unbound DISP1 conformations display distinct Na+site occupancies; these differences, in conjunction with the unusual locations of certain lipids bound to DISP1, suggest a mechanism by which transmembrane Na+flux may power extraction of the lipid-linked Hedgehog signal from the membrane, thus enabling its tissue patterning activity. The Na+-coordinating residues resolved in our DISP1 structures are wholly or partly conserved in some of the other metazoan RND family members, such as PTCH1 and NPC1, suggesting the utilization of Na+flux to power their conformationally-driven activities.
Cloning of Human ABCB11 Gene inE. colirequired the removal of an Intragenic Pribnow-Schaller Box before it’s Insertion into Genomic Safe Harbor AAVS1 Site using CRISPR Cas9
Authors: Vats Nisha , Sanal Madhusudana Girija , Venugopal Senthil Kumar , Taneja Pankaj , Sarin Shiv Kumar
Published: September 2020

Background

Genomic safe harbors are sites in the genome which are safe for gene insertion such that the inserted gene will function properly, and the disruption of the genomic location doesn’t cause any foreseeable risk to the host. The AAVS1 site is the site which is disrupted upon integration of Adeno Associated Virus (AAV) and is considered a ‘safe-harbor’ in human genome because about one third of humans are infected with AAV and so far there is no apodictic evidence that AAV is pathogenic or disruption of AAVS1 causes any disease in man. Therefore, we chose to target AAVS1 site for the insertion of ABCB11, a bile acid transporter which is defective in Progressive Familial Intra Hepatic Cholestasis Type-2 (PFIC-2), a lethal disease of children where cytotoxic bile salts accumulate inside hepatocytes killing them and eventually the patient.

Methods

We used CRISPR Cas9 a genome editing tool to insert ABCB11 gene at AAVS1 site in human cell-lines.

Results

We found that human ABCB11 sequence has a “Pribnow- Schaller Box” which allows its expression in bacteria and expression of ABCB11 protein which is toxic toE. coliand the removal of the same was required for successful cloning. We inserted ABCB11 at AAVS1 site in HEK 293T using CRISPR-Cas9 tool. We also found that ABCB11 protein has similarity withE. coliEndotoxin (Lipid A) Transporter MsbA.

Conclusion

We inserted ABCB11 at AAVS1 site using CRISPR-Cas9, however, the frequency of homologous recombination was very low for this approach to be successful in-vivo (Figure: pictorial abstract).

Pictorial Abstract

ABCB11 gene (which codes the transporter of human bile salts) is targeted to AAVS1 site using a construct which has 5’ and 3’ overhangs which are homologous to the AAVS1 site. A Pribnow box was detected inside ABCB11 gene which allowed the gene to transcribe in E. Coli causing bacterial lysis probably through competitive replacement of a homologous transporter protein in E. Coli (E. coli Endotoxin (Lipid A) Transporter) MsbA, resulting in Lipid A (L) accumulation inside the bacteria.
Drosophila suzukiiavoidance of microbes in oviposition choice
Authors: Sato Airi , Tanaka Kentaro M. , Yew Joanne Y. , Takahashi Aya
Published: September 2020

While the majority ofDrosophilaspecies lay eggs onto fermented fruits, females ofD. suzukiipierce the skin and lay eggs into ripening fruits using their serrated ovipositors. The changes of oviposition site preference must have accompanied this niche exploitation. In this study, we established an oviposition assay to investigate the effects of commensal microbes deposited by conspecific and heterospecific individuals, and showed that presence of microbes on the oviposition substrate enhances egg-laying ofD. melanogasterandD. biarmipes, but discourages that ofD. suzukii. This result suggests that a drastic change has taken place in the lineage leading toD. suzukiiin how females respond to chemical cues produced by microbes. We also found that hardness of the substrate affects the response to microbial growth, indicating that mechanosensory stimuli interact with chemosensory invoked decisions to select or avoid oviposition sites.
Poorly expressed alleles of several human immunoglobulin heavy chain variable (IGHV) genes are common in the human population
Authors: Ohlin Mats
Published: September 2020

Extensive diversity has been identified in the human heavy chain immunoglobulin locus, including allelic variation, gene duplication, and insertion/deletion events. Several genes have been suggested to be deleted in many haplotypes. Such findings have commonly been based on inference of germline repertoire from data sets covering antibody heavy chain encoding transcripts. The inference process operate under conditions that may limit identification of genes transcribed at low levels. The presence of rare transcripts that would indicate the presence of poorly expressed alleles in haplotypes that otherwise appear to have deleted these genes has now been assessed. Alleles IGHV1-2*05, IGHV1-3*02, IGHV4-4*01, and IGHV7-4-1*01 were all identified as being expressed at very low levels from multiple haplotypes, haplotypes that by inference often appeared not to express these genes at all. These alleles harbor unusual sequence variants that may compromise the functionality of the encoded products. Transcripts of two of these alleles to a large degree do not encode a functional product, suggesting that these alleles might be non-functional. It is proposed that the functionality status of immunoglobulin genes should also include assessment of their ability to encode functional protein products.
Saccharomyces cerevisiaeEcm2 Modulates the Catalytic Steps of pre-mRNA Splicing
Authors: van der Feltz Clarisse , Nikolai Brandon , Schneider Charles , Paulson Joshua C. , Fu Xingyang , Hoskins Aaron A.
Published: September 2020

Genetic, biochemical, and structural studies have elucidated the molecular basis for spliceosome catalysis. Splicing is RNA catalyzed and the essential snRNA and protein factors are well-conserved. However, little is known about how non-essential components of the spliceosome contribute to the reaction and modulate the activities of the fundamental core machinery. Ecm2 is a non-essential yeast splicing factor that is a member of the Prp19-related complex of proteins. Cryo-electron microscopy (cryo-EM) structures have revealed that Ecm2 binds the U6 snRNA and is entangled with Cwc2, another non-essential factor that promotes a catalytically active conformation of the spliceosome. These structures also indicate that Ecm2 and the U2 snRNA likely form a transient interaction during 5’ splice site (SS) cleavage. We have characterized genetic interactions between ECM2 and alleles of splicing factors that alter the catalytic steps in splicing. In addition, we have studied how loss of ECM2 impacts splicing of pre-mRNAs containing non-consensus or competing SS. Our results show that ECM2 functions during the catalytic stages of splicing. It facilitates the formation and stabilization of the 1st-step catalytic site, promotes 2nd-step catalysis, and permits alternate 5’ SS usage. We propose that Cwc2 and Ecm2 can each fine-tune the spliceosome active site in unique ways. Their interaction network may act as a conduit through which splicing of certain pre-mRNAs, such as those containing weak or alternate splice sites, can be regulated.
Septal HDAC1 facilitates long-lasting extinction of social fear in male mice
Authors: Bludau Anna , Paul Andreas , Neumann Inga D , Menon Rohit
Published: September 2020

Social anxiety disorder (SAD) is primarily caused by traumatic social events and is characterized by intense fear and avoidance of socially enriched contexts. Formation of such intense social aversion requires learning of the association between trauma and originally neutral social stimuli. Acetylation of histones in response to learning is critical for long-term memory formation. Class I histone deacetylases (HDACs) have emerged as powerful negative regulators of long-term memory formation. However, the lack of clarity in isoform and spatial specificity in HDAC function exists. Utilizing the social fear conditioning (SFC) as well as cued fear conditioning (CFC) paradigms, we aimed to functionally characterize the role of class I HDACs, especially HDAC1, in the regulation of aversive memories in socialversusnon-social contexts. Using cFos expression as a marker of cellular activity, we revealed that the lateral septum (LS) was specifically activated post-acquisition in socially fear conditioned (SFC+) mice. We further measured an increase in activity-inducing HDAC1 phosphorylation at serine residues within the septum of SFC+mice during extinction of social fear. Pharmacological inhibition of HDAC1 within the LS facilitated, while viral overexpression of septal HDAC1 impaired extinction of social fear. Finally, we found that the abovementioned facilitation of extinction memory by HDAC1 inhibition was long-lasting and persists even after 30 days of extinction. Our results show that LS-HDAC1 is a key regulator of long-lasting social fear extinction, which points towards HDAC1 as a potential therapeutic target for SAD.
Limitation of phosphate assimilation maintains cytoplasmic magnesium homeostasis
Authors: Bruna Roberto E. , Kendra Christopher G. , Groisman Eduardo A. , Pontes Mauricio H.
Published: September 2020

Phosphorus (P) is an essential component of several core biological molecules. In bacteria, P is mainly acquired as inorganic orthophosphate (Pi). Once in the cytoplasm, Pi is incorporated into adenosine triphosphate (ATP), which exists primarily as a Mg2+salt. Notably, whereas P is essential, excess of cytosolic Pi hinders growth. Here we demonstrate that cytotoxic effects of excessive Pi uptake result from its assimilation into ATP and subsequent disruption of Mg2+dependent processes. We show thatSalmonella entericacells experiencing cytoplasmic Mg2+starvation restrict Pi uptake, thereby limiting the availability of an ATP precursor. This response prevents excessive ATP synthesis, overproduction of ribosomal RNA, chelation of free cytoplasmic Mg2+and the destabilization of Mg2+-dependent core processes that ultimately hinder bacterial growth and leads to loss of cellular viability. We demonstrate that, even when cytoplasmic Mg2+is not limiting, excessive Pi uptake leads to increased ATP synthesis, depletion of free cytoplasmic Mg2+, inhibition of translation and growth. Our results establish that bacteria must restrict Pi uptake to prevent the depletion of cytoplasmic Mg2+. Furthermore, they provide a framework to understand the molecular basis of Pi cytotoxicity and reveal a regulatory logic employed by bacterial cells to control P assimilation.

Importance

Phosphorus (P) is essential for life. As the fifth most abundant element in living cells, P is required for the synthesis of an array of biological molecules including (d)NTPs, nucleic acids and membranes. Organisms typically acquire environmental P as inorganic phosphate. While essential for growth and viability, excessive intracellular Pi is toxic for both bacteria and eukaryotes. Using the bacteriumSalmonella entericaas a model, we demonstrate that Pi cytotoxicity is manifested following its assimilation into ATP, which acts as a chelating agent for intracellular cations, most notably, Mg2+. These results identify physiological processes disrupted by excessive Pi and elucidate a regulatory logic employed by bacteria to prevent uncontrollable P assimilation.
Characterizing tissue composition through combined analysis of single-cell morphologies and transcriptional states
Authors: Bao Feng , Deng Yue , Wan Sen , Wang Bo , Dai Qionghai , Altschuler Steven J. , Wu Lani F.
Published: September 2020

Advances in spatial transcriptomics technologies enable optical profiling of morphological and transcriptional modalities from the same cells within tissues. Here, we presentmulti-modalstructuredembedding (MUSE), an approach to deeply characterize tissue heterogeneity through analysis of combined image and transcriptional single-cell measurements. We demonstrate that MUSE can discover cellular subpopulations missed by either modality as well as compensate for modality-specific noise. MUSE identified biologically meaningful cellular subpopulations and stereotyped spatial patterning within heterogeneous mouse cortex brain tissues, profiled by seqFISH+ or STARmap technologies. MUSE provides a framework for combining multi-modal single-cell data to reveal deeper insights into the states, functions and organization of cells in complex biological tissues.
Effect of single‐ versus double‐layer uterine closure during caesarean section on postmenstrual spotting (2Close): multicentre, double‐blind, randomised controlled superiority trial
Authors: Stegwee SI , van der Voet LF , Ben AJ , de Leeuw RA , van de Ven PM , Duijnhoven RG , Bongers MY , Lambalk CB , de Groot CJM , Huirne JAF ,   for the 2Close study group
Published: September 2020

Objective

To evaluate whether double‐layer uterine closure after a first caesarean section (CS) is superior compared with single‐layer uterine closure in terms of postmenstrual spotting and niche development in the uterine caesarean scar.

Design

Multicentre, double‐blind, randomised controlled superiority trial.

Setting

Thirty‐two hospitals in the Netherlands.

Population

A total of 2292 women aged ≥18 years undergoing a first CS were randomly assigned to each procedure (1:1): 1144 women were assigned to single‐layer uterine closure and 1148 women were assigned to double‐layer uterine closure.

Methods

Single‐layer unlocked closure and double‐layer unlocked closure, with the second layer imbricating the first.

Main outcome measures

Number of days with postmenstrual spotting during one menstrual cycle 9 months after CS. Secondary outcomes: perioperative and menstrual characteristics; transvaginal ultrasound measurements.

Results

A total of 774 (67.7%) women from the single‐layer group and 770 (67.1%) women from the double‐layer group were evaluable for the primary outcome, as a result of drop‐out and amenorrhoea. The mean number of postmenstrual spotting days was 1.33 (bootstrapped 95% CI 1.12–1.54) after single‐layer closure and 1.26 (bootstrapped 95% CI 1.07–1.45) after double‐layer closure (adjusted mean difference −0.07, 95% CI −0.37 to 0.22, P = 0.810). The operative time was 3.9 minutes longer (95% CI 3.0–4.9 minutes, P < 0.001) and niche prevalence was 4.7% higher (95% CI 0.7–8.7%, P = 0.022) after double‐layer closure.

Conclusions

The superiority of double‐layer closure compared with single‐layer closure in terms of postmenstrual spotting after a first CS was not shown. Long‐term obstetric follow‐up of our trial is needed to assess whether uterine caesarean closure guidelines should be adapted.

Tweetable abstract

Double‐layer uterine closure is not superior for postmenstrual spotting after a first caesarean; single‐layer closure performs slightly better on other outcomes.
Why for feed and not for human consumption? The black soldier fly larvae
Authors: Bessa Leah W. , Pieterse Elsje , Marais Jeannine , Hoffman Louwrens C.
Published: September 2020

With the surge in consumption of insects, the search continues to find ways to increase the popularity of insect‐based products in the Western world. The black soldier fly larvae (BSFL), which is mainly utilized for animal feed, has great potential to provide a sustainable source of nutrients for human food. This review aims to discuss some of the key benefits and challenges of BSFL and their potential role as a food ingredient and/or product for human consumption. Few articles specifically discuss BSFL as a food source, therefore a comprehensive literature search strategy consisted of collecting and evaluating published data about BSFL as animal feed that could be relevant to its use in food. The hurdles that need to be overcome in order to introduce BSFL as a viable food option include safety concerns, technofunctional properties, nutritional aspects, consumer attitudes, and product applications for BSFL.
Impact of the COVID‐19 pandemic on patients with chronic rheumatic diseases: A study in 15 Arab countries
Authors: Ziadé Nelly , el Kibbi Lina , Hmamouchi Ihsane , Abdulateef Nizar , Halabi Hussein , Hamdi Wafa , Abutiban Fatemah , el Rakawi Manal , Eissa Mervat , Masri Basel
Published: September 2020

Aim

To evaluate the impact of the coronavirus disease 2019 pandemic (COVID‐19) on the access to rheumatology care for patients with chronic rheumatic diseases (CRD) in the Arab countries.

Method

A web‐based cross‐sectional survey was designed by the Arab Adult Arthritis Awareness group (AAAA) consisting of 16 rheumatologists representing countries from the Arab League of Associations for Rheumatology (ArLAR) and was validated by the ArLAR scientific committee. The survey was disseminated online through social media and patients' association channels between May 8 and May 22, 2020. The steering committee developed recommendations to improve the care of patients with CRD during the COVID‐19 pandemic.

Results

A total of 2163 patients were included in the analysis; 72% were female; mean age was 40 years (SD 11.9). The Levant, the Gulf, and North Africa contributed almost equally to the sample. The pandemic had a significant negative impact on rheumatology visits in 82% of cases, access to hydroxychloroquine (47%), and chronic medication persistency (28%). The negative impact on rheumatology visits was associated with female gender, country, medication non‐persistency, isolation due to COVID‐19, and impact on mental health. Sixty‐one patients (2.8%) stated that they had COVID‐19, 5% said that a close contact was infected, and 47% were in isolation because of COVID‐19.

Conclusion

The current study highlights the deleterious consequences of the COVID‐19 pandemic on the continuity of rheumatology care. Therefore, an action plan, including establishing a telemedicine platform, securing drug availability, and promoting medication persistence through the appropriate communication channels, is strongly recommended.
Efficacy of abiraterone acetate in castration‐resistant metastatic prostate cancer: A real‐world data analysis
Authors: Oyman Abdilkerim , Başak Mustafa , Özçelik Melike , Özyükseler Deniz Tataroğlu , Işık Selver , Yıldırım Mahmut Emre
Published: September 2020

Purpose

The aim of this study was to evaluate the efficacy of abiraterone in patients with castration‐resistant metastatic prostate cancer.

Materials and methods

We retrospectively analyzed 51 patients with mCRPC treated with abiraterone acetate from January 2014 to August 2018. Clinicopathological information, treatment modalities, treatment responses, and survival times were retrospectively reviewed.

Results

A total of 51 patients who received abiraterone 1000 mg/day + prednisone 10 mg/day between January 2014 and August 2018 were included in the study. Of these patients, 33 (64.7%) had post‐chemotherapy (CT) and 18 (35.3%) had CT‐naive abiraterone receipt. Median overall survival (OS) was 17.3 months (range 9.3‐33.1). Median OS was found to be 12.7 months (range 9.4‐18.3) and 29.4 months (range 9.3‐33.0) in the CT‐naive and post‐CT group, respectively (P = .236). Median radiographic PFS (rPFS) was 10.1 months (range 4.5‐18.4). In the CT‐naive group, rPFS was 10.1 months (IQR 6.0‐14.7) and in the post‐CT group, it was 9.7 months (range 4.0‐18.4) (P = .808). PSA progression‐free survival (PSA‐PFS) was 9.1 months (range 4.6‐13.1). In the CT‐naive group, PSA‐PFS was 7.4 months (range 4.6‐13.4) and in post‐CT, it was 9.1 months (range 4.8‐13.1) (P = .843).

Conclusions

These results show that abiraterone acetate is an effective and reliable agent in real‐life data.
The cytokine profile of menstrual blood
Authors: Crona Guterstam Ylva , Strunz Benedikt , Ivarsson Martin A. , Zimmer Christine , Melin Anna‐Sofia , Jonasson Aino F. , Björkström Niklas K. , Gidlöf Sebastian Brusell
Published: September 2020

Introduction

The menstrual cycle is regulated by a complex interplay between endometrial epithelial cells, endothelial cells, immune cells, and sex hormones. To communicate, cells secrete cytokines that have multiple and diverse effects on recipient cells. Knowledge of how these cells interact in the uterus is insufficient. Menstrual blood is easily accessible and provides a source to study menstrual cycle physiology. This study aimed to determine the cytokine profile in menstrual blood plasma and investigate the differences in cytokine profiles between menstrual and peripheral blood plasma. Several previous studies indicate an improved chance of embryo implantation after endometrial scratching. Consequently, our secondary aim was to compare the menstrual blood cytokine profile before and after luteal phase endometrial scratching.

Material and methods

Nineteen healthy donors collected menstrual blood for the first 24 hours of menstruation in two sequential cycles. Matched peripheral blood was taken at the same time. An endometrial biopsy was performed at cycle day 7‐9 post ovulation in between the two collection times. A Luminex multiplex assay was performed in one batch analyzing a predetermined group of cytokines in plasma.

Results

Peripheral blood plasma and menstrual blood plasma showed substantial significant differences in cytokine profile. In menstrual blood plasma, C5/C5a, interleukin‐6 (IL‐6), IL‐1β, and CXCL8 were detected in high concentrations, whereas IL‐2, IL‐12p70, XCL1/Lymphotactin, and interferon‐γ were low. The most pronounced median differences between menstrual and peripheral blood plasma were found for IL‐6, IL‐1β, and CXCL8. The cytokine profiles of menstrual blood plasma were similar between the individual donors and did not differ over two subsequent cycles. None of the cytokines analyzed in menstrual blood plasma differed significantly before or after luteal phase endometrial scratching (P < .01).

Conclusions

Our results demonstrate that the menstrual blood cytokine profile is distinctly different from peripheral blood plasma and that the inter‐individual difference in menstrual blood cytokine profile in healthy donors is limited and stable over time. The small injury caused by an endometrial biopsy does not change the cytokine profile in the subsequent menstrual cycle. Our study provides new insights into menstrual cycle physiology.
The challenge to define the optimal prophylactic regimen for vitamin K deficiency bleeding in infants
Authors: Lembo Chiara , Buonocore Giuseppe , Perrone Serafina
Published: September 2020

Infants are at risk of vitamin K deficiency that may lead to vitamin K deficiency bleeding (VKDB). Although many vitamin K prophylactic regimens have been developed throughout the years, still cases of late form VKBD may occur. The introduction of combined prophylactic strategy with prolonged oral prophylaxes after the intramuscular dose at birth has showed a decrease of the late severe VKDB incidence. Nevertheless, there is still lack of consensus about the administration scheme after the first dose at birth.

Conclusion

Late form VKBD is not eradicated, and the best prophylactic regimen in term and preterm infants is still an open debate.
Reference interval for albumin‐adjusted calcium based on a large UK population
Authors: Schini Marian , Hannan Fadil M. , Walsh Jennifer S. , Eastell Richard
Published: September 2020

Context

Primary hyperparathyroidism is a common condition and results in hypercalcaemia, especially in older women. Thus, it is critical to obtain a robust estimate for the upper limit of the reference interval for albumin‐adjusted serum calcium in the general population. The current reference interval in use in the UK (Pathology Harmony range, 2.20 to 2.60 mmol/L) was based on a consensus.

Objectives

To establish a reference interval for albumin‐adjusted serum calcium in men and women.

Design

Cross‐sectional study of men and women who did not have chronic kidney disease or vitamin D deficiency; outliers were identified statistically and then rejected and then a 99% reference interval was calculated.

Patients

502 524 men and women aged 40 to 69 years from the UK Biobank Study.

Measurements

Serum total calcium, albumin, 25‐hydroxyvitamin D, estimated glomerular function (eGFR).

Results

We developed an equation for albumin‐adjusted serum calcium and applied it to 178 377 men and women who did not have chronic kidney disease or vitamin D deficiency. We identified 2962 (1.7%) as outliers, and when excluded, we report a 99% reference interval of 2.19 to 2.56 mmol/L (8.76 to 10.24 mg/dL). We found that for older (55‐69 years) and younger women (40‐55 years) the upper limits were 2.59 mmol/L and 2.57 mmol/L and that for all men, the upper limit was 2.55 mmol/L.

Conclusions

We have established an upper limit of the reference range for older women that would identify all high outliers (2.60 mmol/L and above). The upper limit for young women and for men is lower, at 2.57 and 2.55 mmol/L respectively. The current reference interval in use has to be updated and improved based on these findings. These upper limits may prove helpful for identifying hypercalcaemic disorders like primary hyperparathyroidism in clinical practice.
RAR‐related orphan receptor A: One gene with multiple functions related to migraine
Authors: Farahani Sedigheh , Solgi Leila , Bayat Sahar , Abedin Do Atieh , Zare‐Karizi Shohreh , Safarpour Lima Behnam , Mirfakhraie Reza
Published: September 2020

Aims

RAR‐related orphan receptor (RORA) involves in regulation of several biological processes including inflammation and circadian rhythm that probably are involved in migraine pathophysiology. In the current study, the association between RORA rs11639084 and rs4774388 variants and susceptibility to migraine were investigated in a sample of Iranian migraine patients for the first time.

Methods

In a case‐control study including 400 participants, 200 migraineurs and 200 healthy controls, genotyping of RORA rs4774388 and rs11639084 polymorphisms was performed using tetra‐primer amplification refractory mutation system–polymerase chain reaction (TP‐ARMS‐PCR).

Results

The distribution of rs4774388 C/T and T/T genotypes differed significantly between the studied groups. Moreover, an association was observed between rs4774388 and migraine under the recessive mode of inheritance (P = 0.002; OR = 1.89.; CI = 1.25‐2.87). The distribution of rs11639084 alleles and genotypes was not significantly different between migraineurs and healthy controls.

Conclusion

Current results suggest RORA, as a molecular link, may explain inflammation and circadian rhythm dysfunction in migraine. Further studies in different ethnicities are required to confirm the function of RORA in migraine development.