Monitoring the communal incidence of COVID 19 is important for the government and residents of an area to make informed decisions. However, continuous reliance on one means of monitoring might not be ...accurate because of biases introduced by government policies or behaviours of residents. Wastewater surveillance was employed to monitor the distribution of SARS-COV-2 RNA in raw influent wastewater from different wastewater treatment plants serving three Canadian Prairie cities with different population sizes. Data obtained from wastewater are not directly influenced by government regulations or behaviours of individuals. The means of three weekly samples collected using 24 h composite auto-samplers were determined. Viral loads were determined by qPCR, and whole-genome sequencing was used to screen for variants of concern (VOC). The dominant VOCs in the three cities were the same but with different proportions of sub-lineages. Sub-lineages of Delta were AY.12, AY.25, AY.27 and AY.93 in 2021, while the major sub-lineage of Omicron was BA.1 in January 2022, and BA.2 subsequently became a trace-level sub-variant then the predominant VOC. The times that each VOC was first detected varied among cities; however, Saskatoon, with the largest population, was always the first to present new VOCs. Viral loads varied among cities, but there was no direct correlation with population size, possibly because of differences in flow regimes. The population is one of the factors that affect trends in the onset and development of local outbreaks during the pandemic. This might be due to demography or the fact that the larger populations had greater potential for inter- and intra-country migration. Hence, wastewater surveillance data from larger cities can typically be used to indicate what to expect in smaller communities.
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•Omicron supplanted Delta in two weeks.•The largest city was the first to have a new variant of concern.•AY.12, AY.25, AY.27, and AY.93 were the common Delta sublineages.•BA. 1 and BA.1.1 were the consensus sequence in January 2022.•Sub-consensus sequences showed lineages at the trace level.
A switch DNA template was designed to transform proteins into linear DNA strands of 97 nt. The linear DNA was subjected to quantitative polymerase chain reaction (qPCR) to determine the initial copy ...number, which correlated positively with the protein concentration. A restriction endonuclease was used to remove background amplification. Using prostate-specific antigen (PSA) as a protein model, the established method quantified PSA in the range of 10−18-10−13 mol/mL (detection limit = 0.034 pg/mL) with an R2 of 0.974. Good repeatability and specificity of the method were demonstrated. The established method was successfully applied for the quantification of serum PSA levels in women. Significant differences in PSA levels were observed between healthy participants and polycystic ovary syndrome patients.
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•A switch DNA template with specific response to protein biomarker was designed.•The template transformed protein biomarker into linear DNAs of 97 nt.•The linear DNAs were subjected to qPCR for quantification of PSA.
Cellular senescence is a state of irreversibly arrested proliferation, often induced by genotoxic stress 1. Senescent cells participate in a variety of physiological and pathological conditions, ...including tumor suppression 2, embryonic development 3, 4, tissue repair 5–8, and organismal aging 9. The senescence program is variably characterized by several non-exclusive markers, including constitutive DNA damage response (DDR) signaling, senescence-associated β-galactosidase (SA-βgal) activity, increased expression of the cyclin-dependent kinase (CDK) inhibitors p16INK4A (CDKN2A) and p21CIP1 (CDKN1A), increased secretion of many bio-active factors (the senescence-associated secretory phenotype, or SASP), and reduced expression of the nuclear lamina protein LaminB1 (LMNB1) 1. Many senescence-associated markers result from altered transcription, but the senescent phenotype is variable, and methods for clearly identifying senescent cells are lacking 10. Here, we characterize the heterogeneity of the senescence program using numerous whole-transcriptome datasets generated by us or publicly available. We identify transcriptome signatures associated with specific senescence-inducing stresses or senescent cell types and identify and validate genes that are commonly differentially regulated. We also show that the senescent phenotype is dynamic, changing at varying intervals after senescence induction. Identifying novel transcriptome signatures to detect any type of senescent cell or to discriminate among diverse senescence programs is an attractive strategy for determining the diverse biological roles of senescent cells and developing specific drug targets.
•The transcriptome of senescent cells is highly heterogeneous•Senescence transcriptome programs depend on the cell type and stress•Gene expression in senescent cells is temporally dynamic•We identified 55 genes at the core of the senescence-associated transcriptome
The phenotype of senescent cells is highly heterogeneous, but reasons for this variability are poorly understood. Hernandez-Segura et al. identify senescence transcriptome signatures that are strongly associated with specific stresses and cell types and show that the gene expression profiles of various senescence programs are highly dynamic.
External and internal factors are involved in controlling the growth of fishes. However, little is known about the mechanisms by which external factors trigger stimulus signals. This study explored ...the physiological roles of melatonin in the transcription of growth-related genes in the brain and liver of Chrysiptera cyanea, a tropical damselfish with long-day preference. In brain samples of this species collected at 4-h intervals, the transcript levels of arylalkylamine N-acetyltransferase2 (aanat2), the rate-limiting enzyme of melatonin synthesis, and growth hormone (gh) peaked at 20:00 and 00:00, respectively. Concomitantly, the transcript levels of insulin-like growth factors (igf1 and igf2) in the brain and liver were upregulated during the scotophase. Levels of iodothyronine deiodinases (dio2 and dio3), enzymes that convert thyroxine (T4) to triiodothyronine (T3) and reverse T3, respectively, increased in the brain (dio2 and dio3) and liver (dio2) during the photophase, whereas dio3 levels in the liver showed the opposite trend. Fish reared in melatonin-containing water exhibited significant increases in the transcription levels of gh and igf1 in the brain and igf1 in the liver, suggesting that growth in this fish is positively regulated by the GH/IGF pathway on a daily basis. Melatonin treatment also stimulated the transcript levels of dio2 and dio3 in the liver, but not in the brain. Fish consuming pellets containing T3, but not T4, showed significant increases in gh and igf1 in the brain and igf1 and igf2 in the liver, suggesting that the intercellular actions of the TH/IGF pathway have an impact on growth on a daily basis. In summary, IGF synthesis and action in the brain and liver undergo dual regulation by distinct hormone networks, which may also be affected by daily, seasonal, or nutritional factors.
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•Growth-related genes were measured in the brain and liver of damselfish.•Upregulation of igf genes during scotophase was induced by melatonin treatment.•Upregulation of dio2 during photophase was observed in the brain and liver.•Feeding fish with T3 resulted in upregulation of igf genes in the brain and liver.•There are at least two pathways to regulate the expression of igf genes in this fish.
The increase in incidence and geographical expansion of viruses transmitted by the Aedes mosquitoes, such as dengue (DENV) and zika (ZIKV) in the Americas, represents a burden for healthcare systems ...in tropical and subtropical regions. These and other under-detected arboviruses co-circulate in Costa Rica, adding additional complexity to their management due to their shared epidemiological behavior and similarity of symptoms in early stages. Since diagnostics of febrile illness is mostly based on clinical symptoms alone, we gathered acute-phase serum and urine from 399 samples of acute dengue-like cases from two healthcare facilities of Costa Rica, during an outbreak of arboviruses from July 2017 to May 2018, and tested them using molecular and serological methods. The analyses showed that of the clinically presumptive arbovirus cases that were reported, only 39.4% (n=153) of the samples were confirmed positive by RT-PCR to be DENV (DENV (10.3%), CHIKV (0.2%), ZIKV (27.3%), or mixed infections (1.5%). RT-PCR for other alphaviruses and flaviviruses, and PCR for Leptospira sp were negative. Furthermore, to assess flavivirus positivity in post-acute patients, the negative sera were tested against Dengue-IgM. 20% of sera were found positive, confounding even more the definitive number of cases, and emphasizing the need of several distinct diagnostic tools for accurate diagnostics. Molecular characterization of the prM and E genes from isolated viruses revealed that the American/Asian genotype of DENV-2 and the Asian lineage of ZIKV were circulating during this outbreak. Two different clades of DENV-2 American/Asian genotype were identified to co-circulate in the same region and a difference in the platelet and leukocyte count was noted between people infected with each clade, suggesting a putative distinct virulence. Our study sheds light on the necessity for healthcare strategies in managing arbovirus outbreaks, emphasizing the importance of comprehensive molecular and serological diagnostic approaches, as well as molecular characterization. This approach aids in enhancing our understanding of the clinical and epidemiological aspects of arboviral diseases during outbreaks. Our research highlights the need to strengthen training programs for health professionals and the need to increase research-based on laboratory evidence for diagnostic accuracy, guidance, development and implementation of public health interventions and epidemiological surveillance.
Background
In addition to non‐coding RNAs (lncRNAs) and microRNAs (miRNAs), circular RNAs (circRNAs) are endogenous RNAs with various functions, which have recently become a research hotspot. ...CircRNAs are a kind of closed circular RNA molecule widely existing in transcriptomes. Due to lack of free ends, they are not easily cleaved by RNase R, thus avoiding degradation. They are more stable than linear RNAs.
Methods
Data were collected through PubMed. The following search terms were used: “circular RNA,” “circRNA,” “cancer,” “mechanism,” “biogenesis,” “biomarker,” “diagnosis.” Only articles published in English were included.
Results
Most circRNAs express tissue/developmental stage specificity. Moreover, circRNAs are involved in the regulation of a variety of biological activities. In this review, we discuss the formation, classification, and biological functions of circRNAs, especially their molecular diagnostic values in common cancers, including gastric cancer (hsa_circ_002059, circ_LARP4, hsa_circ_0000190, hsa_circ_0000096, circ‐SFMBT2, and circ_PVT1), hepatocellular carcinoma (circ_104075, circRNA_100338, circ_MTO1, and circZKSCAN1), colorectal cancer (hsa_circ_0136666 and hsa_circ_0000523), lung cancer (hsa_circ_0006427, circ_100876, and circ_ABCB10), breast cancer (hsa_circ_0089105, circAGFG1, and circEPSTI1), bladder cancer (circFNDC3B and circTFRC), and esophageal squamous cell carcinoma (circ_100876 and circ‐DLG1).
Conclusion
CircRNAs not only play important roles in tumorigenesis, but also may become new diagnostic biomarkers.
Freshwater harmful cyanobacterial blooms (HCBs) potentially produce excessive cyanotoxins, mainly microcystins (MCs), significantly threatening aquatic ecosystems and public health. Accurately ...predicting HCBs is thus essential to developing effective HCB mitigation and prevention strategies. We previously developed a novel early-warning system that uses cyanotoxin-encoding genes to predict cyanotoxin production in Harsha Lake, Ohio, USA, in 2015. In this study, we evaluated the efficacy of the early-warning system in forecasting the 2016 HCB in the same lake. We also examined potential HCB drivers and cyanobacterial community composition. Our results revealed that the cyanobacterial community was stable at the phylum level but changed dynamically at the genus level over time. Microcystis and Planktothrix were the major MC-producing genera that thrived in June and July and produced high concentrations of MCs (peak level 10.22 μg·L−1). The abundances of the MC-encoding gene cluster mcy and its transcript levels significantly correlated with total MC concentrations (before the MC concentrations peaked) and accurately predicted MC production as revealed by logistic equations. When the Microcystis-specific gene mcyG reached approximately 1.5 × 103 copies·mL−1 or when its transcript level reached approximately 2.4 copies·mL−1, total MC level exceeded 0.3 μg L−1 (a health advisory limit) approximately one week later (weekly sampling scheme). This study suggested that cyanotoxin-encoding genes are promising predictors of MC production in inland freshwater lakes, such as Harsha Lake. The evaluated early-warning system can be a useful tool to assist lake managers in predicting, mitigating, and/or preventing HCBs.
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•Early-warning system based on qPCR/RT-qPCR detection of cyanotoxin-encoding genes•Reoccurrence of a harmful cyanobacterial bloom (HCB) event in Harsha Lake in 2016•Variations of cyanotoxin-encoding genes and cyanotoxin production during the HCB•Significant correlations between microcystin-encoding genes and total microcystins•Cyanotoxin-encoding genes are powerful predictors of cyanotoxin production.
RT-qPCR dissects transcription-based processes but requires reference genes (RGs) for data normalization. This study prospected RGs for mouse macrophages (pMØ) and spleen infected with
. The pMØ were ...infected
with
or vehicle for 4 h. Mice were injected with
(or vehicle) and euthanized 24 h post-injection. The RGs came from a multispecies primer set, from the literature or designed here. The RG ranking relied on GeNorm, NormFinder, BestKeeper, Delta-CT and RefFinder.
-
-
were the most stable RGs for pMØ, albeit RG indexes fine-tuned estimations of cytokine relative expression.
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were the best RGs for spleen but modestly impacted the cytokine relative expression. Hence, mouse models of
require context-specific RGs for RT-qPCR, thus reinforcing its paramount contribution to accurate gene expression profiling.
Reference genes (RGs) were selected for two mouse models of
infection. Five different software were used to select the most stable RGs. The resulting RGs were validated by measuring the relative expression of key cytokine gene transcripts after
infection.
Selection of reference genes for gene expression profiling in mouse models of
infection.
Aims
To assess the removal of viruses through the multiple steps of wastewater treatment in a full‐scale municipal wastewater treatment plant in Alberta, Canada.
Methods and Results
Samples were ...collected after each of the five treatment steps for a period of 16 months. The amount of viruses and their infectivity were analysed using real‐time quantitative PCR (qPCR) and integrated viral cell culture (ICC), respectively. Bacterial indicator Escherichia coli was also tested using membrane filtration. Seven viruses including Norovirus (NoV), Rotavirus (RV), Sapovirus (SaV), Astrovirus (AsV), Adenovirus (AdV), Enterovirus (EV) and JC virus (JCV) were detected in 16 primary effluents in which infectious viruses were present. Different treatment steps showed various efficiencies in virus removal, with membrane ultrafiltration as the most effective at 4·6–7·0 log reduction.
Conclusions
We observed high prevalence of viruses in raw wastewater and different viral reduction after various treatment steps. The discharge of treated wastewater with infectious viruses represents potential risks to human, animal and environmental health.
Significance and Impact of the Study
This study provides a comprehensive assessment of the removal of NoV, RV, SaV, AsV, AdV, EV, JCV and Reovirus from wastewater by current procedures of municipal wastewater treatment and discusses the applicability of various viruses as viral indicators for water quality.
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