Search

Abstract DNA methylation is an epigenetic mark that plays an important role in defining cancer phenotypes, with global hypomethylation and focal hypermethylation at CpG islands observed in tumors. These methylation marks can also be used to define tumor types and provide an avenue for biomarker identification. The homeobox gene class is one that has potential for this use, as well as other genes that are Polycomb Repressive Complex 2 targets. To begin to unravel this relationship, we performed a pan-cancer DNA methylation analysis using sixteen Illumina HM450k array datasets from TCGA, delving into cancer-specific qualities and commonalities between tumor types with a focus on homeobox genes. Our comparisons of tumor to normal samples suggest that homeobox genes commonly harbor significant hypermethylated differentially methylated regions. We identified two homeobox genes, HOXA3 and HOXD10, that are hypermethylated in all 16 cancer types. Furthermore, we identified several potential homeobox gene biomarkers from our analysis that are uniquely methylated in only one tumor type and that could be used as screening tools in the future. Overall, our study demonstrates unique patterns of DNA methylation in multiple tumor types and expands on the interplay between the homeobox gene class and oncogenesis.

Abstract Myometrial stem/progenitor cells (MyoSPCs) have been proposed as the cells of origin for uterine fibroids, but the identity of the MyoSPC has not been well established. We previously identified SUSD2 as a possible MyoSPC marker, but the relatively poor enrichment in stem cell characteristics of SUSD2+ over SUSD2- cells compelled us to find better markers. We combined bulk RNA-seq of SUSD2+/- cells with single cell RNA-seq to identify markers for MyoSPCs. We observed seven distinct cell clusters within the myometrium, with the vascular myocyte cluster most highly enriched for MyoSPC characteristics and markers. CRIP1 expression was found highly upregulated by both techniques and was used as a marker to sort CRIP1+/PECAM1- cells that were both enriched for colony forming potential and able to differentiate into mesenchymal lineages, suggesting that CRIP1+/PECAM1- cells could be used to better study the etiology of uterine fibroids.

Summary: Exploring early embryonic gene expression is challenging due to the rate of development and the limited material available. Here, we present a protocol for ordering Drosophila embryos along a developmental pseudo-time trajectory and determining the sex of the embryos using RNA-seq data. We describe steps for sample collection, RNA isolation, RNA-seq, and RNA-seq data processing. We then detail the establishment of a continuous transcriptome dataset for assessing gene expression throughout early development and in a sex-specific manner.For complete details on the use and execution of this protocol, please refer to Pérez-Mojica et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

A diterpenoid biosynthetic gene cluster (BGC) has been identified in a few species in the Lamiaceae (mint) family, but its origin and evolution remain unclear. Here, the authors report assembly of genomes of three species within the family and reveal the dynamic evolutionary trajectory of the BGC.

Summary: Obese individuals experience low grade inflammation initiated within their adipose tissue. However, the early events that lead to the release of these inflammatory factors from adipose tissue are poorly characterized. To separate glucose effects from lipid effects on adipose tissue, we used an adipose-specific TXNIP knockout model where excess basal glucose influx into adipocytes led to modest increase in adiposity without using high fat diet. We found an uncoupling of two events that are generally presumed to be coregulated: (1) an increase of adipose tissue macrophage (ATM) number; and (2) pro-inflammatory activation of ATMs. These two events are associated with different triggering signals: elevated free fatty acids output and extracellular matrix remodeling with increased ATM number, whereas decreased adiponectin level with activated ATM. This separation reflects non-overlapping pathways regulated by glucose and lipids in adipocytes, and neither group alone is sufficient to elicit the full inflammatory response in adipose tissue.

Summary: While TXNIP (thioredoxin interacting protein) in the plasma membrane and vesicular location is known to negatively regulate cellular glucose uptake by facilitating glucose transporter endocytosis, the function of TXNIP in the nucleus is far less understood. Herein, we sought to determine the function of nuclear TXNIP in vivo, using a new HA-tagged TXNIP knock-in mouse model. We observed that TXNIP can be found in the nucleus of a variety of cells from different tissues including hepatocytes (liver), enterocytes (small intestine), exocrine cells (pancreas), and brown adipocytes (BAT). Further investigations into the role of nuclear TXNIP in BAT revealed that cold stress rapidly and transiently activated HSF1 (heat shock factor 1). HSF1 interaction with TXNIP during its activation is required for optimal HSF1 directed cold shock response in BAT.

Summary: Antibody-secreting cells (ASCs) are key contributors to humoral immunity through immunoglobulin production and the potential to be long-lived. ASC persistence has been recognized in the autoimmune thymus (THY); however, only recently has this population been appreciated in healthy THY tissue. We showed that the young female THY was skewed toward higher production of ASCs relative to males. However, these differences disappeared with age. In both sexes, THY ASCs included Ki-67+ plasmablasts which required CD154(CD40L) signals for their propagation. Single cell RNA-sequencing revealed that THY ASCs were enriched for an interferon responsive transcriptional signature relative to those from bone marrow and spleen. Flow cytometry confirmed that THY ASCs had increased levels of Toll-like receptor 7 as well as CD69 and major histocompatibility complex class II. Overall, we identified fundamental aspects of THY ASC biology which may be leveraged for future in depth studies of this population in both health and disease.

Summary: Brown adipose tissue (BAT) has the ability to burn calories as heat. Utilizing BAT thermogenesis is thus an attractive way to combat obesity. However, the transcriptional network resulting in the lipid synthesis to oxidation shift during thermogenesis is not completely understood. Here, we report the regulation of two master regulators of adipogenesis, peroxisome proliferator-activated receptor gamma (PPARγ) and CCAAT/enhancer-binding protein alpha (C/EBPα), during acute cold stress in BAT. We found PPARγ dissociates from DNA in a fifth of its binding sites and these include Cebpa enhancers, leading to decreased C/EBPα expression. This dissociation requires PPARγ binding to activating ligands and is thus modulated by diet. Meanwhile, PPARα also detaches from DNA, and co-activator PGC1α associates with ERRα as part of a transcriptional network regulating lipid metabolism. Subsequent global replacement of C/EBPα by C/EBPβ and its associated transcriptional machinery is required for upregulation of structural lipid synthesis despite general upregulation of fatty acid oxidation.

Uterine fibroids (leiomyomas) affect Black women disproportionately compared with women of other races and ethnicities in terms of prevalence, incidence, and severity of symptoms. The causes of this racial disparity are essentially unknown. We hypothesized that myometria of Black women are more susceptible to developing fibroids, and we examined the transcriptomic and DNA methylation profiles of myometria and fibroids from Black and White women for comparison. Myometrial samples cluster by race in both their transcriptome and DNA methylation profiles, whereas fibroid samples only cluster by race in the latter. More differentially expressed genes (DEGs) were detected in the Black and White myometrial sample comparison than in the fibroid comparison. Leiomyoma gene set expression analysis identified 4 clusters of DEGs, including a cluster of 24 genes with higher expression in myometrial samples from Black women. One of the DEGs in this group, von Willibrands factor (VWF), was significantly hypomethylated in both myometrial samples from Black women and in all fibroids at 2 CpG probes that are near a putative enhancer site and that are correlated with VWF expression levels. These results suggest that the molecular basis for the disparity in fibroid disease between Black and White women could be found in the myometria before fibroid development and not in the fibroids themselves.

DNA methylation is an epigenetic mark that plays an important role in defining cancer phenotypes, with global hypomethylation and focal hypermethylation at CpG islands observed in tumors. These methylation marks can also be used to define tumor types and provide an avenue for biomarker identification. The homeobox gene class is one that has potential for this use, as well as other genes that are Polycomb Repressive Complex 2 targets. To begin to unravel this relationship, we performed a pan-cancer DNA methylation analysis using sixteen Illumina HM450k array datasets from TCGA, delving into cancer-specific qualities and commonalities between tumor types with a focus on homeobox genes. Our comparisons of tumor to normal samples suggest that homeobox genes commonly harbor significant hypermethylated differentially methylated regions. We identified two homeobox genes, HOXA3 and HOXD10, that are hypermethylated in all 16 cancer types. Furthermore, we identified several potential homeobox gene biomarkers from our analysis that are uniquely methylated in only one tumor type and that could be used as screening tools in the future. Overall, our study demonstrates unique patterns of DNA methylation in multiple tumor types and expands on the interplay between the homeobox gene class and oncogenesis. [ABSTRACT FROM AUTHOR]

Objective: To determine the current insecticide resistance status of Aedes (Ae.) aegypti and Ae. albopictus to four insecticides, namely 0.05% deltamethrin, 0.75% permethrin, 5% malathion and 0.25% pirimiphos–methyl using the World Health Organisation (WHO) susceptibility test kit. Methods: Adult bioassays were carried out using the standard protocol of the World Health Organisation. All F1 generation urban and suburban field strains of Ae. aegypti and Ae. albopictus were tested against pyrethroid and organophosphate insecticides, including the presence of piperonyl butoxide (PBO) in four replicates of 25 non-blood-fed female mosquitoes ranging from 3 to 5 days old. The Vector Control Research Unit (VCRU) laboratory strain served as a reference strain. Results: In this study, 0.05% deltamethrin demonstrated a lower value of knockdown time when 50% of the mosquito population died (KT50) and knockdown time when 95% of the mosquito population died (KT95), which is significantly more effective compared to 0.75% permethrin against adult female Ae. aegypti (urban and suburban) and Ae. albopictus (urban and suburban) (ANOVA, P

Abstract Background Pneumonia is a leading diagnosis for patients requiring critical care. Prediction of the causal organisms has proved challenging, and antibiotic overuse is a burning issue. Objective This study aimed to compare the results of the BioFire FilmArray Pneumonia Panel (PN Panel) to standard diagnostic methods and evaluate its potential impact in an intensive care unit (ICU). Methods This is a retrospective study including adult patients admitted to the ICUs with a diagnosis of pneumonia. The agreement between the PN panel results and the standard diagnostic method was analyzed. Actual and potential adjustments of antimicrobials after the PN panel results were reviewed and compared. Results Two hundred ninety three patients with pneumonia were included. The PN panel identified at least one pathogen in 159 specimens (54.2%), while the standard diagnostic method identified at least one pathogen in 132 specimens (45.0%). Positive percentage agreement between the PN panel and standard‐of‐care culture reached 94%, and negative percentage agreement reached 97%. Among 293 patients, the availability of PN results led to escalation of antimicrobials in 38 (19%) patients and de‐escalation or discontinuation of antimicrobials in 95 (32.4%) patients. Conclusion This pilot study showed the PN panel could provide accurate results on the causative pathogens and detect resistant bacteria. It could reduce the use of broad‐spectrum antibiotics and overall antibiotic use. If the PN panel could be checked at the emergency department, targeted antibiotics could be administered when the patient arrived at the ICU.

Al–Si–Zn–Mg–Fe alloy in as-cast and artificially aged (T6) conditions was friction stir-welded with a traveling speed of 200 mm/min and a rotation speed of 800 rpm. Their microstructure, mechanical and corrosion properties were investigated. α-Al, Si, Mg2Si, MgZn2 and Fe-rich phase were detected in the as-cast and pre-aged Al–Si–Zn–Mg–Fe before and after FSW. The secondary phases in pre-aged Al–Si–Zn–Mg–Fe after FSW are slightly more than those in the as-cast Al–Si–Zn–Mg–Fe after FSW. For the pre-aged Al–Si–Zn–Mg–Fe after FSW, the grains are refined and the dislocation density decreases. Hardness of the pre-aged Al–Si–Zn–Mg–Fe is higher than that of the as-cast one, whereas the hardness of weld nugget zones (NZs) of the both FSWed Al–Si–Zn–Mg–Fe joints is lower than their base materials, and the strength coefficients of both FSWed joints of the as-cast and pre-aged Al–Si–Zn–Mg–Fe are 0.82 and 0.9 respectively. The ultimate tensile strength (UTS) and yield strength (YS) of the pre-aged Al–Si–Zn–Mg–Fe after FSW (UTS = 208.86 MPa and YS = 156.72 MPa) is about 35% higher than that of as-cast Al–Si–Zn–Mg–Fe after FSW (UTS = 159.4 MPa and YS = 112.06 MPa) mainly due to the smaller grain size and more precipitates in the former. The corrosion resistance (CR) of the FSWed joint of the pre-aged Al–Si–Zn–Mg–Fe is the highest, owing to the most refined grain size, highest HAGBs, lowest KAM, increase in the close-packed {111} (α-Al) texture, dispersed particles of secondary phases, followed by the pre-aged Al–Si–Zn–Mg–Fe and then the FSWed joint of the as-cast Al–Si–Zn–Mg–Fe, and the as-cast one is the lowest.

Abstract Background Knee osteoarthritis (KOA) is a common degenerative joint condition that causes disability and pain in the elderly population. The prevalence of KOA among persons aged 63 or above is approximately 30%. Previous studies have reported the positive effects of Tui-na treatment and the Chinese herbal formula Du-Huo-Ji-Sheng Decoction (DHJSD) for KOA treatment. The current study aims to evaluate the add-on therapeutic effect of oral administration of DHJSD on KOA in addition to Tui-na. Methods We conducted a prospective, randomized, controlled clinical trial. Seventy study subjects with KOA were randomly assigned to the treatment and control groups in a 1:1 ratio. Both two groups received eight sessions of Tui-na manipulation for 4 weeks. The DHJSD was only administered to the study subjects in the treatment group. The primary outcome measure was rated using the WOMAC at the end of treatment (4 weeks). Secondary outcomes were assessed using EQ-5D-5L, a health-related quality of life with 5-level EQ-5D version at end of treatment (week 4) and follow-up (week 8). Results No statistically significant difference was found between two groups on WOMAC scores at the end of treatment. The mean WOMAC Pain subscale score was significantly lower in the treatment group than control group at week 8 follow up (mean difference, MD − 1.8, 95% CI − 3.5 to − 0.02, P = 0.048). The mean WOMAC Stiffness subscale score was significantly lower in the treatment group than in the control group at week 2 (MD 0.74, 95% CI 0.05 to 1.42, P = 0.035) and week 8 follow up (MD 0.95, 95% CI 0.26 to 1.65, P = 0.008). The mean EQ-5D index value was significantly improved in the treatment group than in the control group at week 2 (MD 0.17, 95% CI 0.02 to 0.31, P = 0.022). The analysis of WOMAC scores and EQ-5D-5L in both groups showed statistically significant improvement with time. No significant adverse effect was found during the trial. Conclusion DHJSD may have an add-on effect in addition to Tui-na manipulation relieving pain and improving stiffness as well as quality of life (QOL) in patients with KOA. The combined treatment was generally safe and well tolerated. Trial registration The study was registered at the ClinicalTrials.gov (website: https://clinicaltrials.gov/ct2/show/NCT04492670 , registry number: NCT04492670), registered on 30 July 2020.

AbstractThis study combined the principles of multimedia message design with an enclosing-rectangle scaffold to facilitate instruction on deriving the area of basic geometric shapes. A pretest/posttest/delayed-posttest design was used to compare the effectiveness of the enclosing-rectangle scaffold with that of conventional dissection/rearrangement methods among low-achieving seventh-grade students. These fifty-six low-achieving students were averagely assigned to one of two teaching methods. In immediate and delayed testing, an enclosing-rectangle scaffold proved significantly more effective than the conventional approach (posttest: F (1,53) = 16.148, p =

Background: COVID-19 outbreaks in residential care homes for the elderly (RCHEs) and for persons with disability (RCHDs) have caused significant morbidity and mortality during 5th epidemic in Hong Kong. This article reviewed COVID-19 outbreaks situation and estimated the effectiveness of receiving at least two-dose of COVID-19 vaccine in preventing severe outcomes. Methods: To estimate attack rates and vaccination coverage, documentation on COVID-19 infection and their vaccination records of residential care homes (RCH) residents reported between December 31, 2021 and May 31, 2022 were reviewed, and infected cases were follow-up for 4 weeks for severe outcomes or death. Correlation between vaccination coverage against attack rate by types of homes was examined. Infected RCH residents with available information were included in the analysis of vaccine effectiveness against severe outcomes and death. Results: COVID-19 vaccination coverage was low in RCHDs (median 0.46, IQR: 0.24–0.76) and very low in RCHEs (median 0.08, IQR: 0.00–0.19). Higher attack rates were recorded among RCHE residents (median 0.84, IQR: 0.64–0.93) and higher case fatality rate (CFR: 28.1%) than in RCHDs (median 0.58, IQR: 0.31–0.84; CFR: 3.9%). The attack rate decreased when vaccination coverage increased for both RCHEs (ρ = −0.131, p < 0.001) and RCHDs (ρ = −0.333, p < 0.001). Comparing with infected residents who were unvaccinated/vaccinated with one-dose, receiving at least two-dose was estimated to be effective in reducing severe outcomes in 31% and 36% of infected RCHE and RCHD residents respectively; with greater reduction in mortality among RCHD than RCHE residents (54% and 38%, respectively). Vaccine effectiveness of two-dose of BNT162b2 against severe outcomes and death are higher than that of CoronaVac. Conclusions: Increasing COVID-19 vaccination could have significant impact on reducing the risk of COVID-19 outbreaks in RCHs. At least two-dose of COVID-19 vaccine is still effective in reducing severe outcomes and death among infected residents in RCHs during Omicron epidemic.

This study aims to evaluate the suitability and reliability of Process Analytical Tools (PAT) in monitoring milk powder blending processes. The uniformity end point was predicted using a Near-Infrared (NIR) probe, and subsequently validated using offline Fourier Transform Infrared Spectroscopy (FTIR). A standard milk formulation (SMF) made up of 50% lactose, 40% skim milk powder, and 10% whey protein concentrate was used. Additionally, the detection limit of the NIR probe was investigated using vitamin C powder. The average predicted uniformity end point using the inline NIR fixed reference (63.89 ± 2.06 min), and dynamic reference conformity test (63.00 ± 5.25 min) were comparable with the offline FTIR measurement (56.6 ± 0.71 min). A three-component Partial Least Square Regression (PLSR) model was constructed and validated for vitamin C. The detection limit is 0.11%, which is higher than the vitamin C level commonly found in most infant milk formula (0.035%).