Putative Role Research Articles

Prevalence of pediatric acute-onset neuropsychiatric syndrome (PANS) in children and adolescents with eating disorders

BackgroundPediatric obsessive–compulsive disorder (OCD) and eating disorder symptoms frequently overlap, clouding diagnostic certainty and hypothesized etiologic factors. Pediatric acute-onset neuropsychiatric syndrome (PANS) is defined by abrupt emergence of core obsessive–compulsive behaviours and/or food restriction with concurrent, ancillary cognitive and behavioral symptoms. Inflammatory and immune processes have putative roles in both PANS and a related described condition with cardinal obsessive–compulsive or tic symptoms, known as pediatric autoimmune neuropsychiatric disorders associated with streptococcal infection (PANDAS). While prevalence of PANS and PANDAS has been examined in tic, movement disorder and OCD populations, this has not yet been systematically examined in a pediatric eating disorder sample.ObjectivesTo identify the lifetime prevalence of those meeting PANS and/or PANDAS criteria within a pediatric eating disorder cohort.MethodsConvenience sampling method was utilized to select consecutive youth (ages 8–18-years) presenting to an interdisciplinary pediatric eating disorder subspecialty program with a confirmed eating disorder and completed parent-report PANS/PANDAS questionnaire (n = 100). A parent-reported measure was used to establish lifetime prevalence rates for PANS and PANDAS. Descriptive and exploratory comparative analyses were conducted between PANS and non-PANS groups. Continuous measures were analyzed using two-tailed independent sample t-tests and categorical measures were analyzed using two-tailed Fisher’s exact tests.ResultsAmong participants, 52% (n = 52) met PANS criteria and 0% (n = 0) met PANDAS diagnostic criteria. Core, abrupt-onset PANS symptoms included both food restriction and obsessive–compulsive symptoms in 63.5% (n = 33), food restriction only in 25% (n = 13), and obsessive–compulsive symptoms only in 11.5% (n = 6) of participants. In comparison to those who did not meet PANS criteria, those in the PANS subgroup were less likely to be male and more commonly prescribed a selective serotonin reuptake inhibitor medication. Significant group differences did not emerge for onset age, body mass index, eating disorder type or comorbid psychiatric/medical/autoimmune illness.ConclusionLifetime prevalence of symptoms in keeping with PANS diagnostic criteria within a pediatric eating disorder cohort was notably higher than that previously reported in OCD or tic disorder cohorts. The overlap between starvation effects and ancillary PANS symptoms may challenge the practical utility of this putative syndrome within the eating disorder population.

Transcriptomics Analysis Reveals a Putative Role for Hormone Signaling and MADS-Box Genes in Mature Chestnut Shoots Rooting Recalcitrance.

Maturation imposes several changes in plants, which are particularly drastic in the case of trees. In recalcitrant woody species, such as chestnut (Castanea sativa Mill.), one of the major maturation-related shifts is the loss of the ability to form adventitious roots in response to auxin treatment as the plant ages. To analyze the molecular mechanisms underlying this phenomenon, an in vitro model system of two different lines of microshoots derived from the same field-grown tree was established. While juvenile-like shoots root readily when treated with exogenous auxin, microshoots established from the crown of the tree rarely form roots. In the present study, a transcriptomic analysis was developed to compare the gene expression patterns in both types of shoots 24 h after hormone and wounding treatment, matching the induction phase of the process. Our results support the hypothesis that the inability of adult chestnut tissues to respond to the inductive treatment relies in a deep change of gene expression imposed by maturation that results in a significant transcriptome modification. Differences in phytohormone signaling seem to be the main cause for the recalcitrant behavior of mature shoots, with abscisic acid and ethylene negatively influencing the rooting ability of the chestnut plants. We have identified a set of related MADS-box genes whose expression is modified but not suppressed by the inductive treatment in mature shoots, suggesting a putative link of their activity with the rooting-recalcitrant behavior of this material. Overall, distinct maturation-derived auxin sensibility and homeostasis, and the related modifications in the balance with other phytohormones, seem to govern the outcome of the process in each type of shoots.

Mechanism Underlying Naringenin Hypocholesterolemic Effects: Involvement of Estrogen Receptor α Subtype

Naringenin (Nar) is one of major citrus flavonoids predominantly found in grapefruit and orange. In vivo studies have demonstrated Nar potential as a normolipidemic agent capable to reduce circulating cholesterol in hypercholesterolemic rabbits, rats, and patients, suggesting a new role for this molecule in cardiovascular disease prevention. Although Nar cholesterol-lowering effects are known, the underlying mechanisms have not yet been elucidated. Interestingly, Nar binds to the estrogen receptors (ERs), modulating both transcriptional and membrane-initiating signals. Although estrogen and ERs are deeply involved in lipid metabolism, no data are available regarding a putative role of these nuclear receptors as mediators of the hypocholesterolemic effect exerted by Nar. Thus, the aim of this work was to study the involvement of ERs in Nar-induced modulation of cholesterol metabolism. Results obtained in HepG2 cell line demonstrate that Nar can modulate the molecular network of cholesterol homeostasis. However, these effects were only partially dependent on the activity of estrogen receptor α. As a whole, our data highlight new molecular mechanisms by which Nar influences cholesterol metabolism, opening a new scenery about dietary impact on human health.

Two Antenna-Enriched Odorant Binding Proteins in Dioryctria abietella Tuned to General Odorants and Insecticides

The management of forest pests has become a significant challenge, particularly for wood borers, because they spend most of the time in the trunks or cones. The coneworm, Dioryctria abietella, is a representative of cone borers as its larvae feed on the cones of Pinaceae plants. The molecular mechanisms underlying the interactions between this species and host plants or habitats can assist in developing strategies for pest control. In this study, we extended the expression profiles of 32 odorant binding proteins (OBPs) in the reproductive tissues of D. abietella, revealing the detectable transcription of 29 genes. Using two DabiOBPs highly expressed in antennae (DabiOBP5 and DabiOBP14) as targets, six compounds with high affinities (dissociation constants < 13 μM) were identified through a reverse chemical ecology strategy, including insecticides widely used for the control of lepidopteran pests. Of these compounds, a floral volatile β-ionone and a pear-produced ester ethyl-(2E,4Z)-decadienoate may serve as behaviorally active compounds in D. abietella. The strong binding of DabiOBPs to insecticides suggested their involvement in insecticide resistance, reflecting sophisticated detoxification mechanisms of this moth. In the molecular simulations, DabiOBP14 possessed stronger interactions with the six ligands compared to DabiOBP5, in which a few key residues within the binding pockets were involved in the formation of hydrogen bonds. This study provides some valuable reference active compounds for the development of lures or repellents in D. abietella and unravels the putative roles of two antenna-dominant DabiOBPs in the perception of plant-derived odorants and insecticides.

The ChaC1 active site: Defining the residues and determining the role of ChaC1-exclusive residues in the structural and functional stability.

The glutathione degrading enzyme ChaC1 is highly upregulated in several cancers and viral infections making it a potential pharmacological target for cancer therapy. As an enzyme, however, ChaC1 has a relatively high Km (~2 mM) towards its natural substrate, and therefore finding its inhibitors becomes very difficult. Given this limitation, a careful mapping of the active site has become necessary. In the current study, the enzyme-substrate complex was generated by docking glutathione with the modeled hChaC1 structure. Using a combination of in silico and wet lab approaches, the active site residues forming direct interactions with the substrate glutathione were identified and validated. Furthermore, the role of residues exclusively conserved in the ChaC family and forming the surface of the active site were also explored for their putative role in active site stabilization. Mutants of these residues have been analysed for their structural stability and interaction with the substrate through MD simulations and MMGBSA binding energy calculations. These findings were experimentally validated by assessment of their function through in vivo assays in yeast. The experimental evidences along with the molecular modeling suggest that residues 38'YGSL'41, D68, R72, E115, and Y143 are responsible for high affinity binding of hChaC1 with the substrate/inhibitor, whereas the residues exclusive to the ChaC family are required for the structural stability of the enzyme and its active site. Such a characterization of essential active site and conserved residues is significant as a key step toward rational design of novel inhibitors of the ChaC1 enzyme.

Discovery and Visualization of Age-Dependent Patterns in the Diurnal Transcriptome of Drosophila.

Many critical life processes are regulated by input from 24-hour external light/dark cycles, such as metabolism, cellular homeostasis, and detoxification. The circadian clock, which helps coordinate the response to these diurnal light/dark cycles, remains rhythmic across lifespan; however, rhythmic transcript expression is altered during normal aging. To better understand how aging impacts diurnal expression, we present an improved Fourier-based method for detecting and visualizing rhythmicity that is based on the relative power of the 24-hour period compared to other periods (RP24). We apply RP24 to transcript-level expression profiles from the heads of young (5-day) and old (55-day) Drosophila melanogaster, and reveal novel age-dependent rhythmicity changes that may be masked at the gene level. We show that core clock transcripts phase advance during aging, while most rhythmic transcripts phase delay. Transcripts rhythmic only in young flies tend to peak before lights on, while transcripts only rhythmic in old peak after lights on. We show that several pathways, including glutathione metabolism, gain or lose coordinated rhythmic expression with age, providing insight into possible mechanisms of age-onset neurodegeneration. Remarkably, we find that many pathways show very robust coordinated rhythms across lifespan, highlighting their putative roles in promoting neural health. We investigate statistically enriched transcription factor binding site motifs that may be involved in these rhythmicity changes.

Enterovirus-Cardiomyocyte Interactions: Impact of Terminally Deleted Genomic RNAs on Viral and Host Functions.

Group B enteroviruses, including coxsackievirus B3 (CVB3), can persistently infect cardiac tissue and cause dilated cardiomyopathy. Persistence is linked to 5' terminal deletions of viral genomic RNAs that have been detected together with minor populations of full-length genomes in human infections. In this study, we explored the functions and interactions of the different viral RNA forms found in persistently infected patients and their putative role(s) in pathogenesis. Since enterovirus cardiac pathogenesis is linked to the viral proteinase 2A, we investigated the effect of different terminal genomic RNA deletions on 2A activity. We discovered that 5' terminal deletions in CVB3 genomic RNAs decreased the levels of 2A proteinase activity but could not abrogate it. Using newly generated viral reporters encoding nano-luciferase, we found that 5' terminal deletions resulted in decreased levels of viral protein and RNA synthesis in singly transfected cardiomyocyte cultures. Unexpectedly, when full-length and terminally deleted forms were cotransfected into cardiomyocytes, a cooperative interaction was observed, leading to increased viral RNA and protein production. However, when viral infections were carried out in cells harboring 5' terminally deleted CVB3 RNAs, a decrease in infectious particle production was observed. Our results provide a possible explanation for the necessity of full-length viral genomes during persistent infection, as they would stimulate efficient viral replication compared to that of the deleted genomes alone. To avoid high levels of viral particle production that would trigger cellular immune activation and host cell death, the terminally deleted RNA forms act to limit the production of viral particles, possibly as trans-dominant inhibitors. IMPORTANCE Enteroviruses like coxsackievirus B3 are able to initiate acute infections of cardiac tissue and, in some cases, to establish a long-term persistent infection that can lead to serious disease sequelae, including dilated cardiomyopathy. Previous studies have demonstrated the presence of 5' terminally deleted forms of enterovirus RNAs in heart tissues derived from patients with dilated cardiomyopathy. These deleted RNAs are found in association with very low levels of full-length enterovirus genomic RNAs, an interaction that may facilitate continued persistence while limiting virus particle production. Even in the absence of detectable infectious virus particle production, these deleted viral RNA forms express viral proteinases at levels capable of causing viral pathology. Our studies provide mechanistic insights into how full-length and deleted forms of enterovirus RNA cooperate to stimulate viral protein and RNA synthesis without stimulating infectious viral particle production. They also highlight the importance of targeting enteroviral proteinases to inhibit viral replication while at the same time limiting the long-term pathologies they trigger.

Transcriptional profiling of the terpenoid biosynthesis pathway and in vitro tests reveal putative roles of linalool and farnesal in nectarine resistance against brown rot

The most devastating fungal disease of peaches and nectarines is brown rot, caused by Monilinia spp. Among the many plant responses against biotic stress, plant terpenoids play essential protective functions, including antioxidant activities and inhibition of pathogen growth. Herein, we aimed to characterize the expression of terpenoid biosynthetic genes in fruit tissues that presented different susceptibility to brown rot. For that, we performed artificial inoculations with Monilinia laxa at two developmental stages (immature and mature fruit) of two nectarine cultivars (‘Venus’ –mid-early season cultivar - and ‘Albared’ –late season cultivar-) and in vitro tests of the key compounds observed in the transcriptional results. All fruit were susceptible to M. laxa except for immature ‘Venus’ nectarines. In response to the pathogen, the mevalonic acid (MVA) pathway of the ‘Venus’ cultivar was highly induced in both stages rather than the methylerythritol phosphate (MEP) pathway, being the expression of some MEP-related biosynthetic genes [e.g., PROTEIN FARNESYLTRANSFERASE (PpPFT), and 3S-LINALOOL SYNTHASE (PpLIS)] different between stages. In ‘Albared’, both stages presented similar responses to M. laxa for both pathways. Comparisons between cultivars showed that HYDROXYMETHYLGLUTARYL-CoA REDUCTASE (PpHMGR1) expression levels were common in susceptible tissues. Within all the terpenoid biosynthetic pathway, linalool- and farnesal-related pathways stood out for being upregulated only in resistant tissues, which suggest their role in mediating the resistance to M. laxa. The in vitro antifungal activity of linalool and farnesol (precursor of farnesal) revealed fungicidal and fungistatic activities against M. laxa, respectively, depending on the concentration tested. Understanding the different responses between resistant and susceptible tissues could be further considered for breeding or developing new strategies to control brown rot in stone fruit.

RNA sequencing in parthenocarpic chili pepper (Capsicum annuum) fruit reveals genes involved in transcriptional regulation of capsaicinoid biosynthesis genes responsible for the fluctuation of pungency

• RNA-sequencing was used to assess transcriptome changes associated with fluctuation of the pungency in chili pepper fruits. • Pungent fluctuation was associated with changes in gene expressions responsible for metabolic pathways around the biosynthesis of capsaicinoids. • Clustering analysis in genes related to transcriptional regulation was performed using disclosed RNA sequencing data. • The nine genes were noted to exhibit expression patterns similar to those of capsaicinoid biosynthesis genes. • Transcript levels in most of these nine genes were proportional to the pepper pungency levels and are inferred to contribute to the fluctuation of pungency. Chili peppers ( Capsicum spp.) are a popular horticultural crop; the characteristic hot taste of chili peppers is the result of a class of compounds called capsaicinoids. In the horticultural production of such peppers, fluctuations in pungency (changes in capsaicinoid content) are an important issue, given that this fluctuation can cause confusion in the distribution or deterioration of pepper quality. Here, we sought to clarify the mechanism of this variation in pungency, specifically by investigating this variation at the transcriptome level. In the present study, we used the pungency-variable sweet chili pepper ‘Shishito’ ( C. annuum ) and created high-pungency seedless fruits by parthenocarpic treatment. RNA-sequencing analysis then was used to comprehensively compare gene expression in these fruits with that in less-pungent seeded fruits. We firstly profiled differentially expressed genes (DEGs) contributing to metabolic pathways involved with capsaicinoid biosynthesis. Subsequently, we employed clustering analysis (using previously published RNA-sequencing data) to explore changes in the expression of genes encoding putative transcription factors that might serve as regulators of the capsaicinoid biosynthesis genes (CABGs). Specifically, we screened nine DEGs ( CaMYB31, WRKY9, TF62308, TF60920, TF62062, TF75838, ACOT9, TF62804 , and TF64623 ) that have gene ontology related to the regulation of transcription, and found that the expression patterns of these DEGs were similar to those of several CABGs. We also employed quantitative reverse transcriptional polymerase chain reaction to investigate the expression (in the pungency-varied fruits) of these DEGs with putative roles in transcription. Accordingly, we observed that the transcript levels in most of these DEGs correlated positively with the amount of capsaicinoid in the placental septum. Therefore, we postulate that these DEGs with putative transcriptional regulatory roles are involved in capsaicinoid biosynthesis, such that the transcript levels of these DEGs may contribute to the fluctuation of pepper pungency.

Transcriptome - wide bioinformatics analysis of the binding sites of RNA - binding proteins and their putative role in mendelian diseases

Background: Post-transcriptional regulation is the control of gene expression at the RNA level. After produced, the stability and distribution of the different transcripts are regulated by means of RNA-binding proteins (RBPs). Mutations in RNA-binding proteins can cause Mendelian diseases - prominently neuromuscular disorders and cancers. This study determines the interaction between RBPs and target-RNA complexes from public data of the ENCODE project and identifies mutations associated with Mendelian diseases that could disrupt the RBP-RNA interactions. Materials and methods: we performed a transcriptome - wide bioinformatics prediction of the binding sites of RBPs in the human transcriptome from public data of the ENCODE project. Results: The majority (54%) of pathogenic mutation putatively affecting the binding sites of RBPs are located in protein - coding genes and are mainly classified as loss - of - function mutations. Mutations located in the binding sites of RBPs related to RNA processing. For 13 diseases, Familial hypercholesterolemia is the most significant disease with about 40% of mutations in ClinVar database located into the binding sites of RBPs (p=2.3e-65), but congenital hypogonadotropic hypogonadism is the disease with the highest percentage of mutations affecting the binding sites of RBPs (98%, p=2.7e-25). The RBPs most involved in human Mendelian diseases by binding sites-disrupting mutations are YBX3, AQR and PRPF8. Conclusions: A large number of Mendelian diseases are potentially mediated by disease - causing variants that potentially disrupt the binding sites of RBPs. This will provide insight sharper on post - transcriptional mechanisms. Besides, it is useful to know the role of protein - RNA interactome networks in pathologies, thereby serving the treatment of diseases. Key words: bioinformatics analysis, ENCODE project, ClinVar, RNA-binding proteins, Mendelian diseases

Distinct non-coding RNA cargo of extracellular vesicles from M1 and M2 human primary macrophages.

Macrophages are important antigen presenting cells which can release extracellular vesicles (EVs) carrying functional cargo including non-coding RNAs. Macrophages can be broadly classified into M1 'classical' and M2 'alternatively-activated' macrophages. M1 macrophages have been linked with inflammation-associated pathologies, whereas a switch towards an M2 phenotype indicates resolution of inflammation and tissue regeneration. Here, we provide the first comprehensive analysis of the small RNA cargo of EVs from human M1 and M2 primary macrophages. Using small RNA sequencing, we identified several types of small non-coding RNAs in M1 and M2 macrophage EVs including miRNAs, isomiRs, tRNA fragments, piRNA, snRNA, snoRNA and Y-RNA fragments. Distinct differences were observed between M1 and M2 EVs, with higher relative abundance of miRNAs, and lower abundance of tRNA fragments in M1 compared to M2 EVs. MicroRNA-target enrichment analysis identified several gene targets involved in gene expression and inflammatory signalling pathways. EVs were also enriched in tRNA fragments, primarily originating from the 5' end or the internal region of the full length tRNAs, many of which were differentially abundant in M1 and M2 EVs. Similarly, several other small non-coding RNAs, namely snRNAs, snoRNAs and Y-RNA fragments, were differentially enriched in M1 and M2 EVs; we discuss their putative roles in macrophage EVs. In conclusion, we show that M1 and M2 macrophages release EVs with distinct RNA cargo, which has the potential to contribute to the unique effect of these cell subsets on their microenvironment.

Identification of cold shock proteins in Helix pomatia snails: The putative role in overwintering

Identification of cold shock proteins in Helix pomatia snails: The putative role in overwintering

Mechanism of IFT-A polymerization into trains for ciliary transport

Intraflagellar transport (IFT) is the highly conserved process by which proteins are transported along ciliary microtubules by a train-like polymeric assembly of IFT-A and IFT-B complexes. IFT-A is sandwiched between IFT-B and the ciliary membrane, consistent with its putative role in transporting transmembrane and membrane-associated cargoes. Here, we have used single-particle analysis electron cryomicroscopy (cryo-EM) to determine structures of native IFT-A complexes. We show that subcomplex rearrangements enable IFT-A to polymerize laterally on anterograde IFT trains, revealing a cooperative assembly mechanism. Surprisingly, we discover that binding of IFT-A to IFT-B shields the preferred lipid-binding interface from the ciliary membrane but orients an interconnected network of β-propeller domains with the capacity to accommodate diverse cargoes toward the ciliary membrane. This work provides a mechanistic basis for understanding IFT-train assembly and cargo interactions.

Stabilization of DNA fork junctions by Smc5/6 complexes revealed by single-molecule imaging.

SMC complexes play key roles in genome maintenance, where they ensure efficient genome replication and segregation. The SMC complex Smc5/6 is a crucial player in DNA replication and repair, yet many molecular features that determine its roles are unclear. Here, we use single-molecule microscopy to investigate Smc5/6's interaction with DNA. We find that Smc5/6 forms oligomers that dynamically redistribute on dsDNA by 1D diffusion and statically bind to ssDNA. Using combined force manipulation and single-molecule microscopy, we generate ssDNA-dsDNA junctions that mimic structures present in DNA repair intermediates or replication forks. We show that Smc5/6 accumulates at these junction sites, stabilizes the fork, and promotes the retention of RPA. Our observations provide a model for the complex's enrichment at sites of replication stress and DNA lesions from where it coordinates the recruitment and activation of downstream repair proteins.

Lack of Detection of Toxoplasma gondii in Pipistrellus spp. Bats from Densely Cat-Populated Areas of NE Spain.

Toxoplasma gondii infection in healthy animals is often asymptomatic. However, some species with little history of contact with the parasite, such as marsupials and New World primates, present high mortality rates after infection. Despite its potential conservation concern, T. gondii infection in insectivorous bats has received little attention, and its impact on bat populations' health is unknown. To assess the putative role of insectivorous bats in the cycle of T. gondii, samples of three species of bats (Pipistrellus pipistrellus, P. pygmaeus and P. kuhlii) collected between 2019 and 2021 in NE Spain were tested for the presence of the parasite using a qPCR. All tissues resulted negative (0.0% prevalence with 95% CI: [0.0-2.6]) for the presence of T. gondii. Unlike previous studies on insectivorous bats from Europe, Asia and America, the present study suggests that Pipistrellus spp. bats do not play a significant role in the epidemiology of T. gondii in NE Spain. Further studies are encouraged to elucidate both the epidemiology of T. gondii and its potential impact on the health of microchiropteran species in Europe.

Hedging at the rear edge: Intraspecific trait variability drives the trajectory of marginal populations in a widespread boreal tree species

Abstract Rear‐edge populations at the warm margin of species distribution are small, isolated and face environmental conditions at the limit of species bioclimatic envelope. Intraspecific phenotypic variation contributing to the persistence of peripheral populations is expected to become increasingly important under future climate conditions to avoid local extirpation where range shifts lag behind climate change velocity. We investigated the putative role of intraspecific phenotypic variation for the maintenance of rear‐edge populations of fire‐prone jack pine (Pinus banksiana), an obligate pyriscent boreal species. We assessed whether variation in cone serotiny is associated with the population trajectory of marginal stands located south of the boreal biome, in the temperate forest where natural wildfires are infrequent and unpredictable. To this end, we estimated stand‐scale serotiny, minimal age and tree size structure in 26 jack pine stands from the rear edge (n = 17 sites) and the core (n = 9 sites) of the species' range in eastern Canada. On average, rear‐edge jack pine populations are less serotinous albeit more variably compared with range‐core populations where serotiny is more uniformly high. Rear‐edge stands are generally older and display reverse J‐shape tree size structure indicative of a multi‐aged demographic equilibrium, whereas range‐core stands are younger and show a unimodal stand structure depicting a single ageing cohort generally lacking interfire recruitment. Eco‐evolutionary dynamics shifts from a dependency on wildfires in range‐core populations to stands that can regenerate and persist without recurrent fires at the rear edge, where stand‐scale serotiny reaches values below 85%. Synthesis. Unlike range‐core populations, rear‐edge jack pine populations can locally rely on a dual life‐history strategy to ensure both steady recruitment during fire‐free intervals and successful postfire regeneration. This capacity to cope with infrequent and unpredictable fire regime should increase the resilience and resistance of jack pine populations as global changes alter fire dynamics of the boreal forest. More generally, unique intraspecific phenotypic variation in rear‐edge populations contributes to long‐term species persistence in marginal environmental conditions that might scale up with global changes. The conservation of rear‐edge populations and their genetic legacy appears crucial for the resilience of species.

In-Silico Profiling of Mirnas Against Type 2 Diabetes Mellitus and Myocardial Infarction Associated Genes

In-silico profiling of miRNAs against type 2 diabetes mellitus and myocardial infarction associated genes Khadam Hussain, Muhammad Yousaf, Iram Murtaza Signal Transduction Laboratory, Department of Biochemistry, Faculty of Biological Sciences, Quaid-i-Azam University, Islamabad 45320, Pakistan Abstract Type 2 diabetes mellitus (T2DM) and myocardial infarction (MI) are among top ten leading cause of death, worldwide. miRNA based therapeutic approach may prove an icebreaker to overcome the life-threatening complications of MI and T2DM. Mature miRNAs are endogenous single stranded, non-coding, comprised 20 to 22 nucleotides long sequence which involved in gene silencing at post transcriptional level. It has reported that T2DM affected individuals have more chance to develop MI. Therefore, a single therapeutic approach is required to regulate the common underlying molecular pathways. For this, we found T2DM and MI associated genes by using genetic testing registry (GTR) of national center of biotechnology information (NCBI). To further validate the genes data, we did gene enrichment analysis via ShinyGO database. In order to predict targeted miRNAs (8-mer and species conserved) we used TargetScan, miRWalk and miRBase tools. Data analysis shows that T2DM and MI linked genes have some common targeted miRNAs like miR-128-3p /27-3p/ 181-5p/ 132-3p/ 212-3p/ 30-5p/ 101-3p.1/ 101-3p.2/ 9-5p/ 142-3p.1/ 15-5p/ 16-5p/ 195-5p/ 424-5p/ 497-5p/ 155-5p/ 124-3p.1/ 124-3p.2/ 506-3p that may play a putative role to design a most effective therapeutic strategy. Another set of miRNAs like miR-1-3p/ 206/ 27-3p/ 181-5p/ 7-5p and miR-144-3p/ 135-5p/ 142-3p.2 have selected for T2DM and MI respectively that can target more than one gene in each pathological condition. Taken together, our findings suggest most appropriate miRNAs selection for designing better therapeutic strategy against T2DM linked MI. To check the potential of each miRNA against T2DM and MI, more investigation is needed.

Diverse mechanisms associated with cyhalofop-butyl resistance in Chinese sprangletop (Leptochloa chinensis (L.) Nees): Characterization of target-site mutations and metabolic resistance-related genes in two resistant populations.

Resistance of Chinese sprangletop (Leptochloa chinensis (L.) Nees) to the herbicide cyhalofop-butyl has recently become a severe problem in rice cultivation. However, the molecular mechanisms of target-site resistance (TSR) in cyhalofop-butyl-resistant L. chinensis as well as the underlying non-target-site resistance (NTSR) have not yet been well-characterized. This study aimed to investigate cyhalofop-butyl resistance mechanisms using one susceptible population (LC-S) and two resistant populations (LC-1701 and LC-1704) of L. chinensis. We analyzed two gene copies encoding the entire carboxyltransferase (CT) domain of chloroplastic acetyl-CoA carboxylase (ACCase) from each population. Two non-synonymous substitutions were detected in the resistant L. chinensis populations (Trp2027-Cys in the ACCase1 of LC-1701 and Leu1818-Phe in the ACCase2 of LC-1704), which were absent in LC-S. As Trp2027-Cys confers resistance to ACCase-inhibiting herbicides, the potential relationship between the novel Leu1818-Phe mutation and cyhalofop-butyl resistance in LC-1704 was further explored by single-nucleotide polymorphism (SNP) detection. Metabolic inhibition assays indicated that cytochrome P450 monooxygenases (P450s) and glutathione S-transferases (GSTs) contributed to cyhalofop-butyl resistance in specific resistant populations. RNA sequencing showed that the P450 genes CYP71Z18, CYP71C4, CYP71C1, CYP81Q32, and CYP76B6 and the GST genes GSTF11, GSTF1, and GSTU6 were upregulated in at least one resistant population, which indicated their putative roles in cyhalofop-butyl resistance of L. chinensis. Correlation analyses revealed that the constitutive or inducible expression patterns of CYP71C4, CYP71C1, GSTF1, and GSTU6 in L. chinensis were strongly associated with the resistant phenotype. For this reason, attention should be directed towards these genes to elucidate metabolic resistance to cyhalofop-butyl in L. chinensis. The findings of this study improve the understanding of mechanisms responsible for resistance to ACCase-inhibiting herbicides in grass-weed species at the molecular level, thus aiding in the development of weed management strategies that delay the emergence of resistance to this class of pest control products.

OSCA Genes in Bread Wheat: Molecular Characterization, Expression Profiling, and Interaction Analyses Indicated Their Diverse Roles during Development and Stress Response

The hyperosmolality-gated calcium-permeable channels (OSCA) are pore-forming transmembrane proteins that function as osmosensors during various plant developmental processes and stress responses. In our analysis, through in silico approaches, a total of 42 OSCA genes are identified in the Triticum aestivum genome. A phylogenetic analysis reveals the close clustering of the OSCA proteins of Arabidopsis thaliana, Oryza sativa, and T. aestivum in all the clades, suggesting their origin before the divergence of dicots and monocots. Furthermore, evolutionary analyses suggest the role of segmental and tandem duplication events (Des) and purifying selection pressure in the expansion of the OSCA gene family in T. aestivum. Expression profiling in various tissue developmental stages and under abiotic and biotic stress treatments reveals the probable functioning of OSCA genes in plant development and the stress response in T. aestivum. In addition, protein-protein and protein-chemical interactions reveal that OSCA proteins might play a putative role in Ca2+-mediated developmental processes and adaptive responses. The miRNA interaction analysis strengthens the evidence for their functioning in various biological processes and stress-induced signaling cascades. The current study could provide a foundation for the functional characterization of TaOSCA genes in future studies.

Identification of a Novel Pleiotropic Transcriptional Regulator Involved in Sporulation and Secondary Metabolism Production in Chaetomium globosum

Chaetoglobosin A (CheA), a well-known macrocyclic alkaloid with prominently highly antimycotic, antiparasitic, and antitumor properties, is mainly produced by Chaetomium globosum. However, a limited understanding of the transcriptional regulation of CheA biosynthesis has hampered its application and commercialization in agriculture and biomedicine. Here, a comprehensive study of the CgXpp1 gene, which encodes a basic helix-loop-helix family regulator with a putative role in the regulation of fungal growth and CheA biosynthesis, was performed by employing CgXpp1-disruption and CgXpp1-complementation strategies in the biocontrol species C. globosum. The results suggest that the CgXpp1 gene could be an indirect negative regulator in CheA production. Interestingly, knockout of CgXpp1 considerably increased the transcription levels of key genes and related regulatory factors associated with the CheA biosynthetic. Disruption of CgXpp1 led to a significant reduction in spore production and attenuation of cell development, which was consistent with metabolome analysis results. Taken together, an in-depth analysis of pleiotropic regulation influenced by transcription factors could provide insights into the unexplored metabolic mechanisms associated with primary and secondary metabolite production.