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Down syndrome is a genetic-based disorder that results from the triplication of chromosome 21, leading to an overexpression of many triplicated genes, including the gene encoding Dual-Specificity Tyrosine Phosphorylation-Regulated Kinase 1A (DYRK1A). This protein has been observed to regulate numerous cellular processes, including cell proliferation, cell functioning, differentiation, and apoptosis. Consequently, an overexpression of DYRK1A has been reported to result in cognitive impairment, a key phenotype of individuals with Down syndrome. Therefore, downregulating DYRK1A has been explored as a potential therapeutic strategy for Down syndrome, with promising results observed from in vivo mouse models and human clinical trials that administered epigallocatechin gallate. Current DYRK1A inhibitors target the protein function directly, which tends to exhibit low specificity and selectivity, making them unfeasible for clinical or research purposes. On the other hand, antisense oligonucleotides (ASOs) offer a more selective therapeutic strategy to downregulate DYRK1A expression at the gene transcript level. Advances in ASO research have led to the discovery of numerous chemical modifications that increase ASO potency, specificity, and stability. Recently, several ASOs have been approved by the U.S. Food and Drug Administration to address neuromuscular and neurological conditions, laying the foundation for future ASO therapeutics. The limitations of ASOs, including their high production cost and difficulty delivering to target tissues can be overcome by further advances in ASO design. DYRK1A targeted ASOs could be a viable therapeutic approach to improve the quality of life for individuals with Down syndrome and their families.

Abstract Splice modulating antisense oligomers (AOs) are increasingly used to modulate RNA processing. While most are investigated for their use as therapeutics, AOs can also be used for basic research. This study examined their use to investigate internally and terminally truncated proprotein convertase subtilisin/kexin type 9 (PCSK9) protein isoforms. Previous studies have used plasmid or viral-vector-mediated protein overexpression to study different PCSK9 protein isoforms, creating an artificial environment within the cell. Here we designed and tested AOs to remove specific exons that encode for PCSK9 protein domains and produced protein isoforms at more physiologically relevant levels. We evaluated the isoforms’ expression, secretion, and subsequent impact on the low-density lipoprotein (LDL) receptor and its activity in Huh-7 cells. We found that modifying the Cis-His-rich domain by targeting exons 10 or 11 negatively affected LDL receptor activity and hence did not enhance LDL uptake although the levels of LDL receptor were increased. On the other hand, removing the hinge region encoded by exon 8, or a portion of the prodomain encoded by exon 2, have the potential as therapeutics for hypercholesterolemia. Our findings expand the understanding of PCSK9 isoforms and their impact on the LDL receptor and its activity at physiologically relevant concentrations.

Vascular Ehlers–Danlos syndrome or Ehlers–Danlos syndrome type IV (vEDS) is a connective tissue disorder characterised by skin hyperextensibility, joint hypermobility and fatal vascular rupture caused by COL3A1 mutations that affect collagen III expression, homo-trimer assembly and secretion. Along with collagens I, II, V and XI, collagen III plays an important role in the extracellular matrix, particularly in the inner organs. To date, only symptomatic treatment for vEDS patients is available. Fibroblasts derived from vEDS patients carrying dominant negative and/or haploinsufficiency mutations in COL3A1 deposit reduced collagen III in the extracellular matrix. This study explored the potential of an antisense oligonucleotide (ASO)-mediated splice modulating strategy to bypass disease-causing COL3A1 mutations reported in the in-frame exons 10 and 15. Antisense oligonucleotides designed to redirect COL3A1 pre-mRNA processing and excise exons 10 or 15 were transfected into dermal fibroblasts derived from vEDS patients and a healthy control subject. Efficient exon 10 or 15 excision from the mature COL3A1 mRNA was achieved and intracellular collagen III expression was increased after treatment with ASOs; however, collagen III deposition into the extracellular matrix was reduced in patient cells. The region encoded by exon 10 includes a glycosylation site, and exon 15 encodes hydroxyproline and hydroxylysine-containing triplet repeats, predicted to be crucial for collagen III assembly. These results emphasize the importance of post-translational modification for collagen III homo-trimer assembly. In conclusion, while efficient skipping of target COL3A1 exons was achieved, the induced collagen III isoforms generated showed defects in extracellular matrix formation. While therapeutic ASO-mediated exon skipping is not indicated for the patients in this study, the observations are restricted to exons 10 and 15 and may not be applicable to other collagen III in-frame exons.

Abstract Antisense oligomers (AOs) are increasingly being used to modulate RNA splicing in live cells, both for research and for the development of therapeutics. While the most common intended effect of these AOs is to induce skipping of whole exons, rare examples are emerging of AOs that induce skipping of only part of an exon, through activation of an internal cryptic splice site. In this report, we examined seven AO-induced cryptic splice sites in six genes. Five of these cryptic splice sites were discovered through our own experiments, and two originated from other published reports. We modelled the predicted effects of AO binding on the secondary structure of each of the RNA targets, and how these alterations would in turn affect the accessibility of the RNA to splice factors. We observed that a common predicted effect of AO binding was disruption of the exon definition signal within the exon’s excluded segment.

As with the wider Canadian economy, the Covid-19 pandemic has represented an unprecedented shock to agri-food supply chains. Given the importance of Canadian agri-food exports, it is vital that supply chains rebound from the shock and disruptions are minimal. If this occurs, they should be able to respond to opportunities created by the supply chains of international competitors having to deal with Covid-19 in export markets. The demand for food is expected to increase in step with the forecast rise in global population to over 9 billion by 2050. In the next decade alone, total food use for cereals is forecast to grow by 1.2 per cent annually for cereals, 1.7 per cent for animal products and 1.9 per cent for pulses/roots and tubers. Agricultural production to be used as food, feed and in industrial applications will increase by 15 per cent in the next decade (OECD/FAO 2019). Canadian agri-food supply chains are well positioned to meet these future needs so long as they can maintain their resiliency. While the Covid-19 pandemic has not yet run its full course, it is already clear that Canadian agri-food supply chains, both those involved in provisioning the domestic market and those serving export destinations, have proved to be resilient in the face of the pandemic’s shocks and are rapidly returning to the high levels of efficiency they possessed prior to Covid-19. In the domestic market, while there were short run disruptions for some products, there were always sufficient substitutes on the shelves such that there was no threat to food security. As incomes fell, there have been increased calls on foodbanks but these are problems with the wherewithal to purchase, not food availability. Domestic supply chains proved themselves to be robust in dealing with major shocks including the shift to food consumed at home. The international supply chains for agri-food products also proved very resilient and are now well positioned to take advantage of opportunities. Some other countries’ supply chains have not proved as resilient or have suffered from government policy interventions. A number of Canada’s major competitors have imposed export bans, creating market opportunities that Canada could fill. Other competitors have been slower to deal with illness in their workforces. In some importing markets, their domestic supply chains are struggling, creating opportunities for imports from Canada. Due to projected global population growth, the increasing international market for major Canadian agricultural products such as wheat, beef and pork, and canola, are lucrative targets for Canada’s economic recovery and growth. Canada’s future success, however, depends on its agri-food supply chains not being encumbered by reactive regulations that reduce their efficiency, and by supportive policies that ensure producers in the supply chain weather the short-term pain from the Covid-19 crisis.

Oligonucleotides and nucleic acid analogues that alter gene expression are now showing therapeutic promise in human disease. Whilst the modification of synthetic nucleic acids to protect against nuclease degradation and to influence drug function is common practice, such modifications may also confer unexpected physicochemical and biological properties. Gapmer mixed-modified and DNA oligonucleotides on a phosphorothioate backbone can bind non-specifically to intracellular proteins to form a variety of toxic inclusions, driven by the phosphorothioate linkages, but also influenced by the oligonucleotide sequence. Recently, the non-antisense or other off-target effects of 2′ O- fully modified phosphorothioate linkage oligonucleotides are becoming better understood. Here, we report chemistry-specific effects of oligonucleotides composed of modified or unmodified bases, with phosphorothioate linkages, on subnuclear organelles and show altered distribution of nuclear proteins, the appearance of highly stable and strikingly structured nuclear inclusions, and disturbed RNA processing in primary human fibroblasts and other cultured cells. Phosphodiester, phosphorodiamidate morpholino oligomers, and annealed complimentary phosphorothioate oligomer duplexes elicited no such consequences. Disruption of subnuclear structures and proteins elicit severe phenotypic disturbances, revealed by transcriptomic analysis of transfected fibroblasts exhibiting such disruption. Our data add to the growing body of evidence of off-target effects of some phosphorothioate nucleic acid drugs in primary cells and suggest alternative approaches to mitigate these effects.

Staphylococcus aureus is a major cause of bacterial poisoning and spread widely in Iraq. In this study vancomycin resistant S. aureus (VRSA) were isolated from wounds, skin, and nose of human . The isolates were identified by using biochemical tests.Sixty one (72.6%) isolates were identified as S.aureus, followed by CoNS 23 (27.3%) from 250 sample collected. Antibiotic susceptibility was determined by disk diffusion ,the results of the susceptibility test indicated that 59 S. aureus isolates have different levels of resistance to antibiotics.In this study tow methods were used to identify resistant and intermediate resistance to vancomycin: which were Kirby-Bauer disk diffusion and automated system Vitek2 method. Results of Disk diffusion method indicated that (19.6%) isolates were resistant to vancomycin.The results of Vitek2 resistant test for 20 isolates indicated that 9(45%) isolates were resistant to vancomycin, with MIC value of (32 μg / ml); 3(15%) isolates showed intermediate resistant to vancomycin, with MIC value of (4 μg / ml),8(40%) isolates showed sensitive to vancomycin with MIC value of (≤0.5-2 μg / ml).Population analysis profile (PAP) method was uesd to detect Heteroresistant Vancomycin-Intermediate Staphylococcus aureus for the 10 isolates. The results showed that 9 (90%) isolates of S.aureus were resistant to vancomycin , while 1(10%) isolate was sensitive.

IntroductionThis study determined the safety and recommended phase 2 dose (RP2D) of the multikinase inhibitor cabozantinib in combination with trifluridine/tipiracil (FTD/TPI) in refractory metastatic colorectal carcinoma (mCRC).Patients and methodsSingle institution investigator-initiated phase 1 study using 3+3 design. Eligible mCRC patients had received prior standard regimens. Cabozantinib was given orally (p.o.) at 20 mg (dose level [DL] 0) or 40 mg (DL 1) daily on days 1-28, and FTD/TPI p.o. at 35 mg/m2 on days 1-5 and 8-12 every 28 days. Prophylactic growth-factor support was allowed.ResultsFifteen patients were enrolled. Median age 56 years (31-80), male (12/15), ECOG 0/1 = 9/6. Three patients were treated at DL 0 and another nine were treated at DL 1, none exhibiting a DLT. Most common any grade (G) treatment related adverse events (TRAE) were diarrhea (50%), nausea (42%), neutropenia (42%), fatigue (33%), and rash (25%). G3-4 TRAE were neutropenia (25%) and thrombocytopenia, hypokalemia, and weight loss (each 8%). No serious TRAE or G5 were reported. The RP2D was determined to be DL 1. Median PFS was 3.8 months (95% CI 1.9-6.8) and disease control rate was 86.7%.ConclusionThe combination of cabozantinib and FTD/TPI is feasible and tolerable at standard doses with the use of growth factors and showed encouraging clinical activity in refractory mCRC.ClinicaltrialsGOV: NCT04868773.

Polyglutamine (polyQ) ataxias are a heterogenous group of neurological disorders all caused by an expanded CAG trinucleotide repeat located in the coding region of each unique causative gene. To date, polyQ ataxias encompass six disorders: spinocerebellar ataxia types 1, 2, 3, 6, 7, and 17 and account for a larger group of disorders simply known as polyglutamine disorders, which also includes Huntington’s disease. These diseases are typically characterised by progressive ataxia, speech and swallowing difficulties, lack of coordination and gait, and are unfortunately fatal in nature, with the exception of SCA6. All the polyQ spinocerebellar ataxias have a hallmark feature of neuronal aggregations and share many common pathogenic mechanisms, such as mitochondrial dysfunction, impaired proteasomal function, and autophagy impairment. Currently, therapeutic options are limited, with no available treatments that slow or halt disease progression. Here, we discuss the common molecular and clinical presentations of polyQ spinocerebellar ataxias. We will also discuss the promising antisense oligonucleotide therapeutics being developed as treatments for these devastating diseases. With recent advancements and therapeutic approvals of various antisense therapies, it is envisioned that some of the studies reviewed may progress into clinical trials and beyond.

Abstract Integrin alpha-4 (ITGA4) is a validated therapeutic target for multiple sclerosis (MS) and Natalizumab, an antibody targeting ITGA4 is currently approved for treating MS. However, there are severe side effects related to this therapy. In this study, we report the development of a novel DNAzyme that can efficiently cleave the ITGA4 transcript. We designed a range of DNAzyme candidates across various exons of ITGA4. RNV143, a 30mer arm-loop-arm type DNAzyme efficiently cleaved 84% of the ITGA4 mRNA in human primary fibroblasts. RNV143 was then systematically modified by increasing the arm lengths on both sides of the DNAzymes by one, two and three nucleotides each, and incorporating chemical modifications such as inverted-dT, phosphorothioate backbone and LNA-nucleotides. Increasing the arm length of DNAzyme RNV143 did not improve the efficiency however, an inverted-dT modification provided the most resistance to 3′ → 5′ exonuclease compared to other modifications tested. Our results show that RNV143A could be a potential therapeutic nucleic acid drug molecule towards the treatment for MS.

Given school enrollments but in the absence of a student residence census, we present a gravity-like model to infer the residential distribution of enrolled students across various administrative units. Multi-scale analysis of the effects of aggregation across different administrative levels allows for the identification of administrative units with sub-optimally located schools and highlights the challenges in allocating resources. Using this method, we verify that the current scheme of free cross-enrollment across administrative boundaries is needed in achieving universal education in the Philippines.

BackgroundPancreatic ductal adenocarcinoma (PDAC) has a poor prognosis, with a 5-year overall survival rate of 10%. In November 2018, NCCN recommended that all patients with PDAC receive genetic counseling (GC) and germline testing regardless of family history. We hypothesized that patients with PDAC were more likely to be referred for testing after this change to the guidelines, regardless of presumed predictive factors, and that compliance would be further improved following the implementation of a hereditary cancer clinic (HCC).MethodsWe conducted a single-institution retrospective analysis of patients diagnosed with PDAC from June 2017 through December 2021 at University of California, Irvine. We compared rates of genetics referral among patients in different diagnostic eras: the 18-month period before the NCCN Guideline change (pre-NCCN era: June 2017 through November 2018), 14 months following the change (post-NCCN era: December 2018 through January 2020), and 18 months after the creation of an HCC (HCC era: June 2020 through December 2021). Family and personal cancer history, genetics referral patterns, and results of GC were recorded. Data were compared using chi-square, Fisher exact, and multivariate analyses.ResultsA total of 335 patients were treated for PDAC (123 pre-NCCN, 109 post-NCCN, and 103 HCC) at University of California, Irvine. Demographics across groups were comparable. Prior to the guideline changes, 30% were referred to GC compared with 54.7% in the post-NCCN era. After the implementation of the HCC, 77.4% were referred to GC (P

Parkin-type autosomal recessive juvenile-onset Parkinson’s disease is caused by mutations in the PRKN gene and accounts for 50% of all autosomal recessive Parkinsonism cases. Parkin is a neuroprotective protein that has dual functions as an E3 ligase in the ubiquitin–proteasome system and as a transcriptional repressor of p53. While genomic deletions of PRKN exon 3 disrupt the mRNA reading frame and result in the loss of functional parkin protein, deletions of both exon 3 and 4 maintain the reading frame and are associated with a later onset, milder disease progression, indicating this particular isoform retains some function. Here, we describe in vitro evaluation of antisense oligomers that restore functional parkin expression in cells derived from a Parkinson’s patient carrying a heterozygous PRKN exon 3 deletion, by inducing exon 4 skipping to correct the reading frame. We show that the induced PRKN transcript is translated into a shorter but semi-functional parkin isoform able to be recruited to depolarised mitochondria, and also transcriptionally represses p53 expression. These results support the potential use of antisense oligomers as a disease-modifying treatment for selected pathogenic PRKN mutations.

Three isolates of P. aeruginosa were isolated from burnt patients. The ability of these isolates for adhesion and formation of slime layer were tested, the result showed that all isolates were able to adherence on the smooth surface. The sensitivity of P. aeruginosa isolates for antibiotics were tested , all isolates were sensitive to Gentamycin, Piperacillin and Amikacin Ciprofloxacin, and resist to Tetracyclin, Amoxicillin, Cephalexine , Ceftriaxone. Ciprofloxacin and Amikacin were found effective against P. aeruginosa isolates with MIC values of 3.8 μg/ ml for Ciprofloxacin and 0.244 μg/ ml for Amikacin The antibacterial effect of Different concentrations of Aloe vera gel, Sesame Oil and Camphor Oil against P. aeruginosa were determined, Camphor was highly effective with Concentration inhibit bacteria value of 10% followed by Sesame Oil (20%) and Aloe vera gel (>75%). The combinations of Aloe vera gel, Sesame Oil and Camphor Oil and antibiotics (Ciprofloxacin and Amikacin ) showed that the efficacy of the two antibiotics (Ciprofloxacin and Amikacin ) against P.aeroginosa isolates was improved in the presence of Aloe vera gel, Sesame Oil and Camphor Oil. Key words : Aloe vera gel, Sesame Oil , Camphor Oil , Pseudomonas aeruginosa.

The process of pre-mRNA splicing is a common and fundamental step in the expression of most human genes. Alternative splicing, whereby different splice motifs and sites are recognised in a developmental and/or tissue-specific manner, contributes to genetic plasticity and diversity of gene expression. Redirecting pre-mRNA processing of various genes has now been validated as a viable clinical therapeutic strategy, providing treatments for Duchenne muscular dystrophy (inducing specific exon skipping) and spinal muscular atrophy (promoting exon retention). We have designed and evaluated over 5000 different antisense oligonucleotides to alter splicing of a variety of pre-mRNAs, from the longest known human pre-mRNA to shorter, exon-dense primary gene transcripts. Here, we present our guidelines for designing, evaluating and optimising splice switching antisense oligomers in vitro. These systematic approaches assess several critical factors such as the selection of target splicing motifs, choice of cells, various delivery reagents and crucial aspects of validating assays for the screening of antisense oligonucleotides composed of 2′-O-methyl modified bases on a phosphorothioate backbone.

One of the crucial aspects of screening antisense oligonucleotides destined for therapeutic application is confidence that the antisense oligomer is delivered efficiently into cultured cells. Efficient delivery is particularly vital for antisense phosphorodiamidate morpholino oligomers, which have a neutral backbone, and are known to show poor gymnotic uptake. Here, we report several methods to deliver these oligomers into cultured cells. Although 4D-Nucleofector™ or Neon™ electroporation systems provide efficient delivery and use lower amounts of phosphorodiamidate morpholino oligomer, both systems are costly. We show that some readily available transfection reagents can be used to deliver phosphorodiamidate morpholino oligomers as efficiently as the electroporation systems. Among the transfection reagents tested, we recommend Lipofectamine 3000™ for delivering phosphorodiamidate morpholino oligomers into fibroblasts and Lipofectamine 3000™ or Lipofectamine 2000™ for myoblasts/myotubes. We also provide optimal programs for nucleofection into various cell lines using the P3 Primary Cell 4D-Nucleofector™ X Kit (Lonza), as well as antisense oligomers that redirect expression of ubiquitously expressed genes that may be used as positive treatments for human and murine cell transfections.

Abstract Connectivity and its role in the persistence and sustainability of marine metapopulations are attracting increased attention from the scientific community and coastal resource managers. Whether protection should prioritize the connectivity structure or demographic characteristics of a given patch is still unclear. We design a three‐stage population model to analyze the relative importance of sources, sinks, quality and extent of juvenile and adult habitat, and node centralities (eigenvector, degree, closeness, and betweenness) as a basis for prioritizing sites. We use a logistic‐type stage‐structured model to describe the local dynamics of a population with a sessile adult stage and network models to elucidate propagule‐exchange dynamics. Our results show that the coupled states of habitat extent and quality, which determine population carrying capacity, are good criteria for protection strategy. Protecting sites on the basis of sources, sinks, or other centrality measures of connectivity becomes optimal only in limited situations, that is, when larval production is not dependent on the adult population. Our findings are robust to a diverse set of larval pathway structures and levels of larval retention, which indicates that the network topology may not be as important as carrying capacity in determining the fate of the metapopulation. Protecting extensive, good quality habitat can help achieve both conservation and fisheries objectives.

Because the network is an open environment, a lot of care must be taken whentransferring sensitive information especially when related with financial data. This dependson the principals to be trusted which is a problematic and since the first step in networksecurity is the authentication, this paper presents a proposed modal for secure mobilebanking (m-banking) applications using Kerberos (the network authentication protocol).The aim of this paper is to establish a secure communication between the clients andmobile-bank application server in which they can use their mobile phone to securely accesstheir bank accounts, make and receive payments, and check their balances.The integration of smart card into classic Kerberos enhances the security for clientauthentication by storing the cryptographic keys and perform dual factor authentication.Other enhancement to Kerberos is the PKINIT in which the shared key is mapped withpublic- private key. To build a robust client authentication the client uses his/her mobilephone to author Kerberos's messages, process the replay and establish securecommunication with the application server.

The increasing usage of social media for conversations, together with the availability of its data to researchers, provides an opportunity to study human conversations on a large scale. Twitter, which allows its users to post messages of up to a limit of 140 characters, is one such social media. Previous studies of utterances in books, movies and Twitter have shown that most of these utterances, when transcribed, are much shorter than 140 characters. Furthermore, the median length of Twitter messages was found to vary across US states. Here, we investigate whether the length of Twitter messages varies across different regions in the UK. We find that the median message length, depending on grouping, can differ by up to 2 characters.

Conversations reflect the existing norms of a language. Previously, we found that utterance lengths in English fictional conversations in books and movies have shortened over a period of 200 years. In this work, we show that this shortening occurs even for a brief period of 3 years (September 2009-December 2012) using 229 million utterances from Twitter. Furthermore, the subset of geographically-tagged tweets from the United States show an inverse proportion between utterance lengths and the state-level percentage of the Black population. We argue that shortening of utterances can be explained by the increasing usage of jargon including coined words.

rom September 2008 to May 2009, 122 specimens were collected by sterile cotton swaps from patients suffering from eye infection at Ibn Al-Haitham Teaching Eye Hospital, 12 of them (9.83% isolation percentage) were diagnosed as Pseudomonas aeruginosa isolates by using biochemical tests, API 20 E system: they were named (P(1), P(2),……..P(12)). The agglutination sera for the grouping of P. aeruginosa test show that all the isolates belong to the P. aeruginosa strains (6, 9, 12, 16). The result show that P10 isolate have a greater ability to adhesion when all P. aeruginosa isolates were tested on Congo red agar medium and adherence to smooth surfaces. The chemical analysis of crude LPS that was extracted from P10 by using digestive enzyme and hot phenol water method showed that the percentage of carbohydrates was 7.3 and the percentage of protein was 1.3. No nucleic acids were found while the chemical analysis of partially purified LPS that was made by gel-filtration chromatography by using Sephacryl 200 S showed that the percentage of carbohydrates in the partial purified LPS was 19.5% and the percentage of binding proteins was 0.006%. No nucleic acids were found. When the molecular weight of LPS was measured by gel-filtration chromatography by using Sepharose C1- 6B- 200 it was found to be 630957 Dalton. The sera were prepared by using two wild type rabbits (weight 2-2.5 kg). They were injected with five doses over 70 days of different concentrations of partially purified LPS that have been extracted from P. aeruginosa P (10) isolate. One rabbit was used as a control that has been injected with PBS. The titer of antibodies in sera was determined by passive haemagglutination and it was in the immunized rabbit’s sera 160, and the best concentration of LPS was 10µg.

The lipopolysaccharide (LPS) was extracted by using digestive enzyme and hot phenol water method from Pseudomonas aeruginosa P (10) a local isolate from patient suffering from eye infection-and partially purified by gel-filtration chromatography. The sera were prepared in rabbits, they were injected with five doses over 70 days of different concentrations of partially purified LPS. The pathological infection was done by using two groups of female mice; the first group represents the non-immunized group while the second group represents the immunized group. The left and right eyes of the two groups were scratched then the left eyes were infected with P. aeruginosa P (10) while the right eyes serve as a control scratched eye. In the second group that represents the immunized group, the mice were immunized intraperitoneally (i.p.) with sera after and before scratching and infection with P. aeruginosa. The results have shown that the typical pathological scores were observed in the left eyes of non-immunized mice group 72hr. after infection with P. aeruginosa while no pathological scores were observed in the left eyes of immunized mice. This indicated that the immunization with LPS gave a protection against P.aeruginosa corneal infection. These results indicate that immunization with LPS reduces the corneal infection caused by P. aeruginosa.

While the impact of crowding on the diffusive transport of molecules within a cell is widely studied in biology, it has thus far been neglected in traffic systems where bulk behavior is the main concern. Here, we study the effects of crowding due to car density and driving fluctuations on the transport of vehicles. Using a microscopic model for traffic, we found that crowding can push car movement from a superballistic down to a subdiffusive state. The transition is also associated with a change in the shape of the probability distribution of positions from a negatively-skewed normal to an exponential distribution. Moreover, crowding broadens the distribution of cars' trap times and cluster sizes. At steady state, the subdiffusive state persists only when there is a large variability in car speeds. We further relate our work to prior findings from random walk models of transport in cellular systems.

TAS-102 is approved for treatment of refractory metastatic gastroesophageal carcinoma (mGEC). This study sought to determine whether the combination of TAS-102 with irinotecan (TASIRI) was safe and effective in previously treated mGEC. This was a single-arm phase 1b study for patients (pts) with mGEC previously treated with at least one line of fluoropyrimidine and platinum-containing regimen. TAS-102 was given at 25 mg/m2 twice daily on days 1 to 5 with irinotecan 180 mg/m2 on day 1 of a 14-day cycle. The primary endpoint was progression-free survival at 6 months ≥ 35% (PFS-6). 20 Pts were enrolled. The study met its primary endpoint. PFS-6 is 40% (95% CI 19.3-60.0). Median PFS and overall survival are 5.3 months and not reached, respectively. 17 of 20 pts had measurable disease by RECIST criteria. Of the 17, 13 had stable disease and 4 had progressive disease as best response (8 pts had tumor shrinkage  5% of pts were anemia (20%) and neutropenia (10%). 2 serious TRAE were reported: G4 febrile neutropenia (n = 1) and G3 hypotension (n = 1). There was no G5 TRAE. The combination of TASIRI showed encouraging clinical activity with a meaningful improvement in PFS-6 compared to historic controls.

Current guidelines recommend neoadjuvant (NAC) and/or adjuvant chemotherapy for locally advanced gastric cancers (LAGCs). However, the choice and duration of NAC regimen is standardized, rather than personalized to biologic response, despite the availability of several different classes of agents for the treatment of gastric cancer (GC). The current trial will use a tumor-informed ctDNA assay (Signatera™) and monitor response to NAC. Based on ctDNA kinetics, the treatment regimen is modified. This is a prospective single center, single-arm, open-label study in clinical stage IB-III GC. ctDNA is measured at baseline and repeated every 8 weeks. Imaging is performed at the same intervals. The primary end point is the feasibility of this approach, defined as percentage of patients completing gastrectomy.

Cholangiocarcinoma (CCA) is a heterogenous group of malignancies originating in the biliary tree, and associated with poor prognosis. Until recently, treatment options have been limited to surgical resection, liver-directed therapies, and chemotherapy. Identification of actionable genomic alterations with biomarker testing has revolutionized the treatment paradigm for these patients. However, several challenges exist to the seamless adoption of precision medicine in patients with CCA, relating to a lack of awareness of the importance of biomarker testing, hurdles in tissue acquisition, and ineffective collaboration among the multidisciplinary team (MDT). To identify gaps in standard practices and define best practices, multidisciplinary hepatobiliary teams from the University of California (UC) Davis and UC Irvine were convened; discussions of the meeting, including optimal approaches to tissue acquisition for diagnosis and biomarker testing, communication among academic and community healthcare teams, and physician education regarding biomarker testing, are summarized in this review.

Children with developmental concerns in Australia continue to experience inequitable healthcare and service-related delays, even when diagnostic risk is identified. This study sought to explore service and demographic pathway factors leading up to autism spectrum disorder (ASD) assessment, including value of screening measures applied at triage. Following a trial of centralised intake for referred young children with suspected ASD, observational, retrospective pathway data was explored using bivariate and regression analyses. The mean age of 159 children referred with autism symptoms was 3.6 years, and 64% were diagnosed with ASD. Service allocation was associated with diagnosis, whilst screening tool results were not. Improved pathways are needed to limit wasted waiting times and direct each child to needs-based services.

Fungi with prominent hairlike rhizomorphs, aerial habit, sparse small sporophores, and usually saprotrophic biology are commonly known as 'horsehair or spider fungi'. The earliest descriptions of them, from the mid-late 1800s, were published from Australian material under the names Marasmius crinis-equi and M. equicrinis. For the original 'horsehair fungus', we review this early historical material, briefly explore the changes in the species concept over time, and investigate the nomenclatural tangle of potentially competing names. Our analysis of morphological and molecular data for over 60 collections across eastern Australia shows that material labelled as M. crinis-equi forms part of a complex of at least three closely related species and that this name has been misapplied both within Australia and internationally. An epitype is nominated for an updated concept of M. crinis-equi, and the closely related taxa, M. tropicus sp. nov. and M. kabakada sp. nov., are described. Two more distantly related Australian taxa to which the name M. crinis-equi has been misapplied, M. perumbilicatus sp. nov. and M. argillaceus sp. nov., are also described to further stabilise the concept of the authentic horsehair fungus M. crinis-equi. [ABSTRACT FROM AUTHOR]

Background and Purpose: This study aimed to assess the validity and reliability of the Dutch Eating Behavior Questionnaire for Children (DEBQ-C) among 6- to 8-year-old Black girls. Methods: Investigators of this cross-sectional online survey of 112 Black mother–daughter dyads used principal component analysis (PCA) to assess evidence of the DEBQ-C's construct validity. Pearson's product-moment correlations were also used to compare the DEBQ-C's subscale scores to an independent measure of eating behaviors. A comparison of the subscale scores between participants with favorable and unfavorable eating behaviors was used to evaluate discriminant validity. coefficient α examined the DEBQ-C's internal consistency reliability. Results: Principal component analysis established evidence of validity with three subscales. Significant correlations were found between the independent measure of eating behavior and the emotional (r = −.25, p =.01) and external subscales (r = −.31, p =.001). Participants increased emotional (t = 2.53; df = 76; p =.007) and external (t = 3.98; df = 99; p <.001) eating scores had unfavorable eating behavior scores. Conclusions: Psychometric results demonstrate questionable support for the construct validity of two of the DEBQ-C subscales (emotional and restrained eating) and the reliability of the DEBQ-C questionnaire among 6- to 8-year-old Black girls. The questionnaire's three subscales, emotional, external, and restrained eating, can be used to examine eating behaviors. [ABSTRACT FROM AUTHOR]

Primary-school-aged children and adolescents with autism spectrum disorder (ASD) are reported to engage in lower levels of moderate-to-vigorous physical activity (MVPA) compared to typically developing (TD) children (Jones et al. in PLoS ONE, 12(2):1-23, 2017). Levels of MVPA in young children with ASD remain unclear. This study aimed to investigate MVPA in 4-to-7-year-old children with (n = 37) and without (n = 40) ASD, to determine if MVPA is related to ASD diagnosis; and examine correlates to better inform interventions. Results indicated children with ASD engage in the same levels of MVPA as TD children. Future studies need to further explore MVPA in children with ASD over time to uncover when the divergence in MVPA levels occur and what factors may be associated.

Vascular Ehlers–Danlos syndrome or Ehlers–Danlos syndrome type IV (vEDS) is a connective tissue disorder characterised by skin hyperextensibility, joint hypermobility and fatal vascular rupture caused by COL3A1 mutations that affect collagen III expression, homo-trimer assembly and secretion. Along with collagens I, II, V and XI, collagen III plays an important role in the extracellular matrix, particularly in the inner organs. To date, only symptomatic treatment for vEDS patients is available. Fibroblasts derived from vEDS patients carrying dominant negative and/or haploinsufficiency mutations in COL3A1 deposit reduced collagen III in the extracellular matrix. This study explored the potential of an antisense oligonucleotide (ASO)-mediated splice modulating strategy to bypass disease-causing COL3A1 mutations reported in the in-frame exons 10 and 15. Antisense oligonucleotides designed to redirect COL3A1 pre-mRNA processing and excise exons 10 or 15 were transfected into dermal fibroblasts derived from vEDS patients and a healthy control subject. Efficient exon 10 or 15 excision from the mature COL3A1 mRNA was achieved and intracellular collagen III expression was increased after treatment with ASOs; however, collagen III deposition into the extracellular matrix was reduced in patient cells. The region encoded by exon 10 includes a glycosylation site, and exon 15 encodes hydroxyproline and hydroxylysine-containing triplet repeats, predicted to be crucial for collagen III assembly. These results emphasize the importance of post-translational modification for collagen III homo-trimer assembly. In conclusion, while efficient skipping of target COL3A1 exons was achieved, the induced collagen III isoforms generated showed defects in extracellular matrix formation. While therapeutic ASO-mediated exon skipping is not indicated for the patients in this study, the observations are restricted to exons 10 and 15 and may not be applicable to other collagen III in-frame exons. [ABSTRACT FROM AUTHOR]

Down syndrome is a genetic-based disorder that results from the triplication of chromosome 21, leading to an overexpression of many triplicated genes, including the gene encoding Dual-Specificity Tyrosine Phosphorylation-Regulated Kinase 1A (DYRK1A). This protein has been observed to regulate numerous cellular processes, including cell proliferation, cell functioning, differentiation, and apoptosis. Consequently, an overexpression of DYRK1A has been reported to result in cognitive impairment, a key phenotype of individuals with Down syndrome. Therefore, downregulating DYRK1A has been explored as a potential therapeutic strategy for Down syndrome, with promising results observed from in vivo mouse models and human clinical trials that administered epigallocatechin gallate. Current DYRK1A inhibitors target the protein function directly, which tends to exhibit low specificity and selectivity, making them unfeasible for clinical or research purposes. On the other hand, antisense oligonucleotides (ASOs) offer a more selective therapeutic strategy to downregulate DYRK1A expression at the gene transcript level. Advances in ASO research have led to the discovery of numerous chemical modifications that increase ASO potency, specificity, and stability. Recently, several ASOs have been approved by the U.S. Food and Drug Administration to address neuromuscular and neurological conditions, laying the foundation for future ASO therapeutics. The limitations of ASOs, including their high production cost and difficulty delivering to target tissues can be overcome by further advances in ASO design. DYRK1A targeted ASOs could be a viable therapeutic approach to improve the quality of life for individuals with Down syndrome and their families. [ABSTRACT FROM AUTHOR]

This research focused on the challenges and coping mechanisms of Non-Social Studies major teachers assigned to teach Araling Panlipunan in Bulacan, Philippines. This, therefore, makes the pressing call for comprehensive content knowledge, effective assessment strategies, collaborative approaches to teaching, and continuous professional development in order to fill the gap in expertise in teaching Philippine history, culture, and society. Key findings revealed that teachers belonging to the Non-Social Studies major often experience problems in their teaching brought about by insufficient subject-specific knowledge, which consequently affects their confidence and accuracy in content delivery. In this regard, it is proposed in the study a program on "Capacitating the Competence of the Non-Social Studies Major Teachers." This program seeks to enhance teaching competencies with focused interventions that equip teachers with necessary competencies and knowledge. These recommendations include a vertical professional learning community, teaching aids bank, and focused seminars and training programs. The study reinforces continuous learning and institutional support as ways to educational excellence and calls for future research into th impact of advanced degrees on teaching effectiveness. [ABSTRACT FROM AUTHOR]