Binding Cassette Research Articles

Effects of uranium mining on soil bacterial communities and functions in the Qinghai-Tibet plateau

The microecological effects of plateau uranium mining are still unknown. In this study, we used 16S rRNA high-throughput sequencing to analyze the impact of plateau uranium mining on the microbial diversity and community structure of tailings soil, tunnel soil, and soil at different depths in an open pit. The results showed that uranium mining significantly reduced soil microbial community richness and diversity indicators, including Chao1, Pielou evenness, and Shannon index (P < 0.05). Uranium mining activities significantly reduced the abundance of RB41, Vicinamidactaceae, and Nitrospira (P < 0.05). Interestingly, the abundance of Thiobacillus, Sphingomonas, and Sulfuriferula significantly increased in the soil samples from various environments and depths during uranium mining (P < 0.05). Beta diversity analysis found that uranium mining resulted in the differentiation of soil microbial communities. Functional enrichment analysis found that uranium mining resulted in the functional enrichment of DNA binding response regulator, DNA helicase, methyl-accepting chemotaxis protein, and Helicase conserved C-terminal domain, whereas cell wall synthesis, nonspecific serine/threonine protein kinase, RNA polymerase sigma-70 factor, and ATP binding cassette transporter were significantly affected by uranium mining (P < 0.05). In addition, we also found that different uranium mining environments and soil depths enriched diverse microbial populations and functions to cope with the environmental pressures that were elicited by uranium mining, including Gaiella, Gemmatimonas, Lysobacter, Pseudomonas, signal transformation histidine kinase, DNA-directed DNA polymerase, and iron complex outer membrane receptor protein functions (P < 0.05). The results have enhanced our understanding of the impact of uranium mining on plateau soil microecological stability and the mechanism of microbial response to uranium mining activities for the first time and aided us in screening microbial strains that can promote the environmental remediation of uranium mining in plateaus.

Tumour suppressor ABCA8 inhibits malignant progression of colorectal cancer via Wnt/β-catenin pathway

BackgroundColorectal cancer (CRC) is one of the most commonly diagnosed malignant tumours of the digestive tract, and new therapeutic targets and prognostic markers are still urgently required. However, the role and molecular mechanisms of ATP binding cassette subfamily A member 8 (ABCA8) in CRC remain unclear. MethodsDatabases and clinical specimens were analysed to determine the expression level of ABCA8 in CRC. Colony formation, CCK-8 and Transwell assays were conducted to determine cell proliferation, viability, migration and invasion. Flow cytometry was used to detect cell cycle progression and apoptosis. Western blot and rescue experiments were performed to determine the specific mechanisms of action of ABCA8. ResultsABCA8 expression is dramatically down-regulated in CRC tissues and cell lines. Ectopic expression of ABCA8 induced apoptosis and cell cycle arrest in vitro, inhibited cell growth, suppressed migration and invasion, reversed epithelial-mesenchymal transition and suppressed xenograft tumour growth and metastasis in vivo. Mechanistically, ABCA8 inhibited CRC cell proliferation and metastasis through the Wnt/β-catenin signalling pathway, both in vitro and in vivo. ConclusionWe verified that ABCA8 inhibits the malignant progression of CRC through the Wnt/β-catenin pathway. This newly discovered ABCA8-Wnt-β-catenin signalling axis is probably helpful in guiding the clinical diagnosis and treatment of CRC.

Integrative transcriptomic and network pharmacology analysis reveals the neuroprotective role of BYHWD through enhancing autophagy by inhibiting Ctsb in intracerebral hemorrhage mice.

In this study, we aimed to combine transcriptomic and network pharmacology to explore the crucial mRNAs and specific regulatory molecules of Buyang Huanwu Decoction (BYHWD) in intracerebral hemorrhage (ICH) treatment. C57BL/6 mice were randomly divided into three groups: sham, ICH, and BYHWD. BYHWD (43.29g/kg) was administered once a day for 7days. An equal volume of double-distilled water was used as a control. Behavioural and histopathological experiments were conducted to confirm the neuroprotective effects of BYHWD. Brain tissues were collected for transcriptomic detection. Bioinformatics analysis were performed to illustrate the target gene functions. Network pharmacology was used to predict potential targets for BYHWD. Next, transcriptomic assays were combined with network pharmacology to identify the potential differentially expressed mRNAs. Immunofluorescence staining, real-time polymerase chain reaction, western blotting, and transmission electron microscopy were performed to elucidate the underlying mechanisms. BYHWD intervention in ICH reduced neurological deficits. Network pharmacology analysis identified 203 potential therapeutic targets for ICH, whereas transcriptomic assay revealed 109 differentially expressed mRNAs post-ICH. Among these, cathepsin B, ATP binding cassette subfamily B member 1, toll-like receptor 4, chemokine (C-C motif) ligand 12, and baculoviral IAP repeat-containing 5 were identified as potential target mRNAs through the integration of transcriptomics and network pharmacology approaches. Bioinformatics analysis suggested that the beneficial effects of BYHWD in ICH may be associated with apoptosis, animal autophagy signal pathways, and PI3K-Akt and mTOR biological processes. Furthermore, BYHWD intervention decreased Ctsb expression levels and increased autophagy levels in ICH. Animal experiments in combination with bioinformatics analysis confirmed that BYHWD plays a neuroprotective role in ICH by regulating Ctsb to enhance autophagy.

ABC transporter subfamily B1 as a susceptibility determinant of Bombyx mori larvae to Cry1Ba, Cry1Ia and Cry9Da toxins

ATP binding cassette (ABC) transporters are a diverse family of transmembrane proteins. Specific subfamily members expressed in the lepidopteran midgut can act as susceptibility determinants for several insecticidal Bt Cry proteins. However, the susceptibility determinants to many Cry toxins still remain unclear. Therefore, we knocked out a series of ABC transporters that are highly expressed in the midgut of Bombyx mori larvae by transcription activator-like effector nuclease (TALEN)-mediated gene editing, and the lineages that became resistant to Cry toxins were searched by toxin overlay bioassay. As a result, the B. mori ABC transporter subfamily B1 (BmABCB1) knockout lineage showed 19.17-fold resistance to Cry1Ba, 876.2-fold resistance to Cry1Ia, and 29.1-fold resistance to Cry9Da, suggesting that BmABCB1 is the determinant of susceptibility to these toxins. BmABCC2 and BmABCC3 have been shown to be susceptibility determinants based on their function as receptors. Therefore, we next heterologously expressed these ABC transporters in HEK293T cells and performed a cell swelling assay to examine whether these molecules could exert receptor functions. As a result, BmABCB1-expressing cells showed swelling response to Cry1Ia and Cry9Da, and cells expressing PxABCB1, which is the Plutella xylostella ortholog of BmABCB1, showed swelling for Cry1Ba, suggesting that ABCB1 is a susceptibility determinant by functioning as a receptor to these toxins. Furthermore, in order to clarify how high binding affinity is based on receptor function, we performed surface plasmon resonance analysis and found that each KD of Cry1Ba, Cry1Ia, and Cry9Da to BmABCB1 were 7.69 × 10−8 M, 2.19 × 10−9 M, and 4.17 × 10−6 M respectively.

Decaffeinated coffee and green tea extract inhibit foam cell atherosclerosis by lowering inflammation and improving cholesterol influx/efflux balance through upregulation of PPARγ and miR-155

Background Foam cells are markers of atherosclerosis and characterise advanced atherosclerotic plaque, stimulated by inflammation caused by high lipid levels in macrophages. The combination of decaffeinated coffee and green tea extract (DCGTE) has been suggested to have a role in foam cell inhibition. Objective We investigated the inhibiting role of DCGTE against foam cell formation, through modulation of the inflammation process and cholesterol metabolism in macrophage colony stimulating factor- (M-CSF) and oxidized low-density lipoprotein (oxLDL)-exposed macrophages. Methods Coffee and green tea were extracted by filtration and infusion respectively, and underwent decaffeination using active carbon and blanching methods, respectively. Cells were administered 160/160 and 320/320μg/ml of DCGTE. Foam cell formation was observed using a light microscope after staining with Oil Red O (ORO), and the accumulation of lipids in macrophages with ELISA. Observations of lipid influx and efflux were determined through semiquantitative cluster differentiation 36 (CD36) and ATP binding cassette transporter A1 (ABCA1) expression through immunofluorescence. The inflammation process was quantified using inflammatory/anti-inflammatory markers, e.g., tumor necrosis factor α (TNFα) and interleukin 10 (IL10) with ELISA. Peroxisome proliferator activated response γ (PPARγ) expression and activity were assessed with PCR and ELISA, respectively. The expression of microRNA 155 (miR-155) was examined using qPCR. Results DCGTE at the above concentrations tended to reduce foam cell numbers, significantly inhibited lipid accumulation (p=0.000), reduced CD36 expression (p=0.000) and TNFα secretion (p=0.000) in Raw264.7 exposed to M-CSF 50ng/ml and oxLDL 50μg/ml. PPARγ expression (p=0.00) and activity (p=0.001), miR-155 relative expression (p=0.000), and IL10 production (p=0.000) also increased. Conclusion DCGTE lowered foam cell numbers, possibly through attenuation of the inflammatory process and improvement of lipid/efflux mechanisms in M-CSF and oxLDL-stimulated Raw264.7 cells, via upregulation of PPARγ and miR-155. Our results suggest DCGTE may help prevent atherosclerosis-based diseases.

Maltose-Derivatized Fluorescence Turn-On Imaging Probe for Bacteria Detection.

We report a maltose-derivatized fluorescence turn-on imaging probe, Mal-Cz, to detect E. coli and Staphylococci. The fluorescence turn-on is achieved through an intramolecular C-H insertion reaction of the perfluoroaryl azide-functionalized carbazole to give a fluorescent product. Confocal fluorescence microscopy confirmed the successful uptake of Mal-Cz by E. coli and Staphylococci upon photoactivation. The Mal-Cz probe could selectively detect E. coli and S. epidermidis in the presence of P. aeruginosa and M. smegmatis without interference from these bacteria. Both the photoactivation and bacteria detection can be accomplished using a hand-held UV lamp at 365 nm, with the limit of detection of 103 CFU/mL by the naked eye. Mal-Cz could also be used to detect E. coli and S. epidermidis spiked in milk by the naked eye under a hand-held UV lamp. The uptake of Mal-Cz requires metabolically active bacteria: the uptake was reduced in stationary phase bacteria and was diminished in bacteria that were killed by heating or treating with antibiotics or sodium azide. The uptake decreased with increasing concentration of added free maltose, indicating that Mal-Cz hijacked the maltose uptake pathways. In E. coli, the maltose transport systems, including maltoporin LamB, maltose binding protein MBP, and the maltose ATP binding cassette (ABC) transporter MalFGK2, are all critical for the transport of Mal-Cz. The uptake was diminished in the deletion mutants ΔLamB, ΔMalE, ΔMalF, and ΔMalK.

Asymmetric conformations and lipid interactions shape the ATP-coupled cycle of a heterodimeric ABC transporter

Here we used cryo-electron microscopy (cryo-EM), double electron-electron resonance spectroscopy (DEER), and molecular dynamics (MD) simulations, to capture and characterize ATP- and substrate-bound inward-facing (IF) and occluded (OC) conformational states of the heterodimeric ATP binding cassette (ABC) multidrug exporter BmrCD in lipid nanodiscs. Supported by DEER analysis, the structures reveal that ATP-powered isomerization entails changes in the relative symmetry of the BmrC and BmrD subunits that propagates from the transmembrane domain to the nucleotide binding domain. The structures uncover asymmetric substrate and Mg2+ binding which we hypothesize are required for triggering ATP hydrolysis preferentially in one of the nucleotide-binding sites. MD simulations demonstrate that multiple lipid molecules differentially bind the IF versus the OC conformation thus establishing that lipid interactions modulate BmrCD energy landscape. Our findings are framed in a model that highlights the role of asymmetric conformations in the ATP-coupled transport with general implications to the mechanism of ABC transporters.

Abstract 16831: Deficiency of Cholesterol Efflux Pathways in Smooth Muscle Cells Enhances Vasoconstriction but Does Not Affect Atherosclerosis

Smooth muscle cells (SMCs) regulate blood flow distribution via vasoconstriction mediated by α-adrenergic receptors (α-ARs). Plasma membrane cholesterol may affect α 1 -AR signaling, but consequences for SMC-mediated vasoconstriction are unclear. Cholesterol loading promotes SMC to macrophage transition in vitro , which may enhance atherosclerotic plaque vulnerability. However, the role of ATP Binding Cassette A1 and G1 (ABCA1/G1)-cholesterol efflux pathways in SMC-mediated vasoconstriction and atherogenesis remains poorly understood. We generated mice with SMC-specific Abca1/g1 deficiency on the low-density lipoprotein receptor deficient ( Ldlr -/- ) background by crossbreeding Abca1 fl/fl Abcg1 fl/fl Ldlr -/- mice with Myh11-Cre ERT2 transgenic mice and feeding them tamoxifen-diet. To induce SMC cholesterol accumulation and atherogenesis, we fed Myh11-Cre ERT2 Abca1 fl/fl Abcg1 fl/fl Ldlr -/- and Myh11-Cre ERT2 Ldlr -/- mice Western-type diet (WTD) for 16 weeks. Combined SMC-Abca1/g1 deficiency increased vasoconstriction in aortic rings induced by the α 1 -AR agonist phenylephrine, with reversal by methyl-β-cyclodextrin, substantiating its cholesterol-dependency. Unexpectedly, SMC - Abca1/g1 deficiency induced urinary bladder enlargement by &gt;20-fold, resembling bladder outlet obstruction (BOO). This was reversed by the α 1 -AR antagonist tamsulosin, indicating its dependence on SMC constriction. Moreover, SMC - Abca1/g1 deficiency decreased SMC markers and increased macrophage- and fibroblast-markers in the bladder wall, enhancing collagen deposition, consistent with SMC transdifferentiation. However, after 16 weeks WTD, SMC - Abca1/g1 deficiency did not affect atherosclerotic lesion size, fibrous cap thickness, necrotic core, collagen, or macrophage content, suggesting that SMCs in atherosclerotic plaques were not affected. This may be due to low Abca1/g1 expression in intimal SMCs. We uncover a new role of SMC cholesterol efflux pathways in suppressing α 1 -AR mediated vasoconstriction and bladder SMC transdifferentiation, decreasing BOO. Our data may provide a mechanistic link for the association between BOO and diabetes in humans, particularly because diabetes is associated with decreased cholesterol efflux.

Two ABCI family transporters, OsABCI15 and OsABCI16, are involved in grain-filling in rice

Seed development is critical for plant reproduction and crop yield, with panicle seed-setting rate, grain-filling, and grain weight being key seed characteristics for yield improvement. However, few genes are known to regulate grain filling. Here, we identify two adenosine triphosphate (ATP)-binding cassette (ABC)I-type transporter genes, OsABCI15 and OsABCI16, involved in rice grain-filling. Both genes are highly expressed in developing seeds, and their proteins are localized to the plasma membrane and cytosol. Interestingly, knockout of OsABCI15 and OsABCI16 results in a significant reduction in seed-setting rate, caused predominantly by the severe empty pericarp phenotype, which differs from the previously reported low seed-setting phenotype resulting from failed pollination. Further analysis indicates that OsABCI15 and OsABCI16 participate in ion homeostasis and likely export ions between filial tissues and maternal tissues during grain filling. Importantly, overexpression of OsABCI15 and OsABCI16 enhances the seed-setting rate and grain yield in transgenic plants and decreases ion accumulation in brown rice. Moreover, the OsABCI15/16 orthologues in maize exhibit a similar role in kernel development, as demonstrated by their disruption in transgenic maize. Therefore, our findings reveal the important roles of two ABC transporters in cereal grain filling, highlighting their value in crop yield improvement.

The role of ATP binding cassette (ABC) transporters in breast cancer: Evaluating prognosis, predicting immunity, and guiding treatment

ABSTRACT Breast cancer is currently the most prevalent form of cancer worldwide. Nevertheless, there remains limited clarity regarding our understanding of the tumor microenvironment and metabolic characteristics associated with it. ATP-binding cassette (ABC) transporters are the predominant transmembrane transporters found in organisms. Therefore, it is essential to investigate the role of ABC transporters in breast cancer. Transcriptome data from breast cancer patients were downloaded from the TCGA database. ABC transporter-related genes were obtained from the Genecards database. By LASSO regression, ABC-associated prognostic signature was constructed in breast cancer. Subsequently, immune microenvironment analysis was performed. Finally, cell experiments were performed to verify the function of ABCB7 in the breast cancer cell lines MDA-MB-231 and MCF-7. Using the ABC transporter-associated signature, we calculated a risk score for each breast cancer patient. Patients with breast cancer were subsequently categorized into high-risk and low-risk groups, utilizing the median risk score as the threshold. Notably, patients in the high-risk group exhibited significantly worse prognosis (P<0.05). Additionally, differences were observed in terms of immune cell infiltration levels, immune correlations, and gene expression of immune checkpoints between the two groups. Functional experiments conducted on breast cancer cell lines MDA-MB-231 and MCF-7 demonstrated that ABCB7 knockdown significantly diminished cell activity, proliferation, invasion, and migration. These findings emphasize the significance of understanding ABC transporter-mediated metabolic and transport characteristics in breast cancer, offering promising directions for further research and potential therapeutic interventions.

Comparative Transcriptome and Metabolome Analyses Revealed Quality Difference between Beauty Tea Processed through Indoor Withering and Outdoor Solar Withering.

Withering is the first processing procedure of beauty tea, and there are few reports on the impact of withering methods on the quality of beauty tea and its regulatory mechanisms. Through comparison of fresh tea leaves (FT) with the leaves at Indoor natural Withering 18h (IWT-18) and Outdoor Solar Withering 6h (OWT-6), which were collected at the end of the two withering processes, 17282 and 13984 differentially expressed genes (DEGs) were screened and 267 and 154 differential metabolites (DMs) were identified, respectively. Coexpression network revealed that a large number of DEGs and DMs were enriched in phenylpropanoid, flavonoid, and adenosine Triphosphate binding (ABC) transporter pathways, and the number of DMs and DEGs in IWT-18 vs FT exceeded that in OWT-6 vs FT. Both withering methods promoted a significant increase in content of phenylalanine and upregulation of β-glucoside (BGLU) expression in the phenylpropanoid metabolism pathway. The five proanthocyanidins related to theaflavins in the flavonoid synthesis pathway was more significant accumulation in FT vs IWT-18 than in FT vs OWT-6. Meanwhile, both withering methods can affect the ABC transporter pathway to promote the accumulation of amino acids and their derivatives, but different withering methods affect different ABC transporter families. Outdoor withering with more severe abiotic stress has a greater impact on the ABCG family, while indoor withering has a more significant effect on the ABCC family. Sensory evaluation results showed that the dry tea of IWT-18 was slightly better than that of OWT-6 because of longer withering time and more thorough substance transformation. In conclusion, the formation of honey flavor in beauty tea may be closely related to the DEGs and DMs in these three pathways. Our research provides theoretical data support for further revealing the mechanism of quality formation during the withering process of beauty tea. This article is protected by copyright. All rights reserved.

Escherichia coli utilizes multiple peptidoglycan recycling permeases with distinct strategies of recycling.

Bacteria produce a structural layer of peptidoglycan (PG) that enforces cell shape, resists turgor pressure, and protects the cell. As bacteria grow and divide, the existing layer of PG is remodeled and PG fragments are released. Enterics such as Escherichia coli go to great lengths to internalize and reutilize PG fragments. E. coli is estimated to break down one-third of its cell wall, yet only loses ~0 to 5% of meso-diaminopimelic acid, a PG-specific amino acid, per generation. Two transporters were identified early on to possibly be the primary permease that facilitates PG fragment recycling, i) AmpG and ii) the Opp ATP binding cassette transporter in conjunction with a PG-specific periplasmic binding protein, MppA. The contribution of each transporter to PG recycling has been debated. Here, we have found that AmpG and MppA/Opp are differentially regulated by carbon source and growth phase. In addition, MppA/Opp is uniquely capable of high-affinity scavenging of muropeptides from growth media, demonstrating that AmpG and MppA/Opp allow for different strategies of recycling PG fragments. Altogether, this work clarifies environmental contexts under which E. coli utilizes distinct permeases for PG recycling and explores how scavenging by MppA/Opp could be beneficial in mixed communities.

Expression analysis of ABCA1 in type 2 diabetic Pakistani patients with and without dyslipidemia and correlation with glycemic index and lipid profile

Diabetes Mellitus type II, earlier considered as an endocrinological disorder is now more regarded as an inflammatory disorder along with lipid aberrations. It demands for regular monitoring, healthy dietary habits and lifestyle modification. This study was focused on gene expression of ATP binding cassette protein 1 (ABCA1) in diabetic dyslipidemia patients in comparison with control groups of only diabetics and healthy individuals. Blood samples and data were collected from recruited 390 patients who were further divided into three groups (130 each). Glycemic index and lipid profile was assessed. Delta Delta Ct method was used that revealed downregulation of the studied gene more in diabetic dyslipidemia patients as compared to only diabetics and healthy controls. The Ct values of ABCA1 were associated with glycemic index and lipid profile using Pearson’s correlation. A negative correlation with fasting blood sugar and a positive correlation with HbA1cwas observed in only diabetics group. While in diabetic dyslipidemia and normal healthy controls, a negative correlation was found with both. As far as the lipid profile is concerned a positive correlation was observed among only diabetics with whole lipid profile. In diabetics with dyslipidemia, a negative correlation with all parameters except the TAGs was observed. A positive correlation with all except HDL was observed in healthy controls. The Ct values and fold change were compared among diseased and healthy individuals by applying independent t test. The cycle threshold in only diabetics was p = 0.000018 and in diabetic dyslipdemia individuals was p = 0.00251 while fold change in only diabetics (p = 0.000230) and in diabetics with dyslipidemia (p = 0.001137) was observed to be as statistically significant.

THU490 Role Of Cholesterol Efflux Pump ABCA1 In Macrophages And Subsequent Modulation Of The Tumor Microenvironment

Abstract Disclosure: S.V. Bendre: None. N. Krawczynska: None. S. Bhogale: None. A. Das Gupta: None. A. Nelczyk: None. H. Vidana Gamage: None. S. Han: None. E. Tajkhorshid: None. S. Sinha: None. W. Cho: None. E.R. Nelson: None. Breast cancer constitutes about 30% of all new female cancers each year. Although there has been an overall decrease in death rates, mortalities associated with recurrent metastatic cancer remain high. Hence, development of targeted therapeutics against metastatic breast cancer is of utmost importance. Although breast cancer is generally considered immunologically “cold”, clinical studies have shown that immunotherapy has potential to improve patient outcomes. One of the challenges of immunotherapy is overcoming the highly immune suppressive tumor microenvironment. Myeloid cells, such as macrophages, which are abundant within the tumor micro-environment, play a key role in cancer cell survival, progression, and metastasis. Several aspects of cholesterol metabolism and regulation have been implicated in breast cancer progression. ABCA1 (ATP Binding Cassette subfamily A member 1) is a membrane bound cholesterol efflux protein, which facilitates cholesterol transport across, as well as between, membrane leaflets. Although implicated in macrophage biology, little is known about how ABCA1 impacts macrophage function in terms of tumor biology. Therefore, the goal of the current work is to determine the effect of altering ABCA1 within macrophages and its subsequent effect on the tumor microenvironment. Intriguingly, our analysis of publicly available transcriptomes indicates that ABCA1 expression within breast tumors is associated with good prognosis, supporting a role for ABCA1 in tumor progression. Important aspects of macrophage function are efferocytosis, antigen presentation, interaction with and activation of T cells, and promotion of angiogenesis. Therefore, we investigated each of these aspects under conditions where ABCA1 expression was manipulated. Our results indicate that knocking down ABCA1 in macrophages decreased the activation and expansion of T cells. Those T cells that were activated, had a decreased anti-cancer cytotoxic response. Over expression of ABCA1 had the opposite effects, supporting our conclusion that ABCA1 in macrophages is critical for normal T cell activation and function. Furthermore, we are finding that ABCA1 is involved in several other aspects of macrophage biology with relevance to tumor progression. Specifically, knockdown of ABCA1 increased the efficiency of efferocytosis, increased angiogenesis, and decreased macrophage infiltration into breast cancer organoids. All these activities suggest that the loss of ABCA1 shifts macrophages into a functional state that is immune-suppressive and pro-tumorigenic. In summary, our results to date indicate that ABCA1 expression within macrophages reduces their pro-tumor functions. Therefore, it will be important to explore this biology in the hopes of developing ABCA1 as a therapeutic target to shape the tumor microenvironment into being less permissive to tumor growth. Presentation: Thursday, June 15, 2023

Fatty acid binding protein 5 regulates docetaxel sensitivity in taxane-resistant prostate cancer cells.

Prostate cancer is a leading cause of cancer-related deaths in men in the United States. Although treatable when detected early, prostate cancer commonly transitions to an aggressive castration-resistant metastatic state. While taxane chemotherapeutics such as docetaxel are mainstay treatment options for prostate cancer, taxane resistance often develops. Fatty acid binding protein 5 (FABP5) is an intracellular lipid chaperone that is upregulated in advanced prostate cancer and is implicated as a key driver of its progression. The recent demonstration that FABP5 inhibitors produce synergistic inhibition of tumor growth when combined with taxane chemotherapeutics highlights the possibility that FABP5 may regulate other features of taxane function, including resistance. Employing taxane-resistant DU145-TXR cells and a combination of cytotoxicity, apoptosis, and cell cycle assays, our findings demonstrate that FABP5 knockdown sensitizes the cells to docetaxel. In contrast, docetaxel potency was unaffected by FABP5 knockdown in taxane-sensitive DU145 cells. Taxane-resistance in DU145-TXR cells stems from upregulation of the P-glycoprotein ATP binding cassette subfamily B member 1 (ABCB1). Expression analyses and functional assays confirmed that FABP5 knockdown in DU145-TXR cells markedly reduced ABCB1 expression and activity, respectively. Our study demonstrates a potential new function for FABP5 in regulating taxane sensitivity and the expression of a major P-glycoprotein efflux pump in prostate cancer cells.

Transcriptomic and proteomic analyses of Mycobacterium tuberculosis strains isolated from tuberculous meningitis patients.

Tuberculous meningitis (TBM) is caused by the dissemination of Mycobacterium tuberculosis (MTB) from the primary site of infection to the central nervous system. However, the bacterial factors associated with the pathogenesis of TBM remain unclear. This study employed transcriptomic and proteomic methods to comprehensively analyze the changes in genes and proteins and their associated pathways in MTB strains isolated from cerebrospinal fluid (CSF) of TBM and sputum of pulmonary TB (PTB) cases. Five MTB strains were subjected to OMICs (transcriptomic and proteomic) analysis. Among five MTB strains, two were isolated from CSF and sputum samples of the same patient with PTB and TBM infections, one from the sputum of a different PTB patient, and a strain obtained from the CSF of another TBM patient. H37Rv was used as a reference strain. The reliability of transcriptomic results was validated by real time polymerase chain reaction with selected genes from 100 MTB isolates (CSF, 50 and sputum, 50). The transcriptomic study revealed that overlapping differentially expressed genes of MTB strains isolated from TBM patients showed featured enrichment in benzoate degradation, lysine degradation, tryptophan metabolism, fatty acid degradation, ATP binding cassette transporters, microbial metabolism in diverse environments, biosynthesis of antibiotics, and metabolic pathways. Eleven genes were upregulated, and four were downregulated in MTB strains isolated from TBM compared to PTB. From proteomic analysis, we identified three candidate proteins belonging to plasminogen binding proteins (PBP) (enolase, dnaK, and isocitrate lyase 1) that were significantly upregulated in MTB strains isolated from TBM. Overall, the transcriptomic and proteomic analyses provided an important base for understanding the unique feature of TBM pathogenesis. To the best of our knowledge, this is the first report highlighting the importance of PBPs on TBM pathogenesis.

301 Measuring ATPase activity of adenosine triphosphate–binding cassette subfamily C member 4, lamprey CFTR, and human CFTR using antimony-phosphomolybdate assay

301 Measuring ATPase activity of adenosine triphosphate–binding cassette subfamily C member 4, lamprey CFTR, and human CFTR using antimony-phosphomolybdate assay

Recurrent Harlequin Ichthyosis in a Family: A Case Report

Introduction: Harlequin ichthyosis is a severe form of erythrodermic ichthyosis that manifests in a distinctive and distressing appearance at birth. The newborn baby is covered in thick, plate-like scales with distorted surface features, resulting in limited mobility. Fortunately, this condition is rare, with an estimated incidence of approximately 1 in 300,000 births. It is inherited in an autosomal recessive manner and is caused by a mutation in the adenosine triphosphate binding cassette subfamily A member 12 (ABCA12) gene. This mutation affects lipid transport, and the activity of ABCA12 is crucial to produce long-chain ceramides, which are essential for the development of a normal skin barrier. Methods: We present a case report of a newborn who presented with thick, firmly adherent dense plaques covering the entire body, accompanied by deep fissures that extended into the dermis. The facial features included severe ectropion, conjunctival chemosis, eclabium, tethered ears, and a rudimentary nose. In addition, there was flexion contracture of the arms, legs, and digits, along with hypoplastic nails. Results: The baby experienced respiratory difficulty, poor sucking, and a severe cutaneous infection. Unfortunately, the baby’s condition deteriorated and passed away on the 20th day after delivery. Conclusion: Previously considered universally fatal, harlequin ichthyosis now has reported cases of survivors, although it remains a challenging condition. It is crucial to provide parents and family with comprehensive information about the implications of the condition. They should be encouraged to explore all available options and receive psychological support throughout the process. It is essential for the neonatal intensive care unit staff to be trained in providing specialized care for these babies. In addition, parents should be offered the option of antenatal counseling, particularly in cases where there is a previously affected sibling or a history of consanguinity.

Effects of Quercetin Nanoemulsion on Cholesterol Efflux and MicroRNA-33/34a Expression in the Liver of Mice Fed with a High-Cholesterol Diet.

Quercetin is a flavonoid widely present in plants; despite its beneficial physiological activity, it exhibits considerably low bioavailability. Nanoemulsion technology is used for improving the bioavailability of lipophilic phenolic compounds. This study aimed to investigate the potential effects of quercetin nanoemulsion (QN) on regulating the microRNA (miR)-33/34a pathway involved in cholesterol efflux in the liver of mice fed with a high-cholesterol (HC) diet. Subsequently, C57BL/6J mice were divided into four groups and fed a normal chow diet, HC diet supplemented with 1% cholesterol and 0.5% cholic acid, or HC diet supplemented with 0.05% QN or 0.1% QN for 6 weeks. Serum and hepatic lipid profiles were assayed using commercial enzymatic kits. Gene expression and miR levels were quantified using real-time quantitative reverse transcription polymerase chain reaction, and adenosine monophosphate-activated protein kinase (AMPK) activity was measured using an AMPK Kinase Assay kit. QN supplementation improved serum and liver lipid profiles. QN upregulated the mRNA levels of adenosine triphosphate (ATP)-binding cassette subfamily A1, ATP-binding cassette subfamily G1, and scavenger receptor class B type 1, which are related to cholesterol efflux. In the QN group, the hepatic AMPK activity increased, whereas miR-33, and miR-34a expression levels decreased. These results suggest that QN may enhance cholesterol efflux, at least partly through modulating AMPK activity and miR-33/34a expression in the liver.

Role of ATP Binding Cassette Subfamily A Member 1 (ABCA1) in Chemoradiotherapy-induced Renal Injury

Radiation therapy (RT) alone or combined with chemotherapy reduces cholesterol efflux and causes chronic kidney disease (CKD). There is no treatment for CKD except renal replacement strategies such as dialysis or transplantation. We hypothesize that cancer treatment-induced ABCA1 deficiency leads to dysregulation of cholesterol metabolism, making podocytes susceptible and cancerous cells resistant to treatment-induced injuries. To quantify the cell index of LNCaP, 786-0, and Podcytes were grown in an e-plate reader (Roche) for 24 h and then were treated with either enzalutamide (10µM) or sunitinib (1.0 µM) or cyclophosphamide (10µM) with or without RT (4Gy) or ABCA1 inducer (liver-X-receptors agonist (GW3965; Sigma; 10µM). Podocyte cholesterol efflux assay was measured using NBD-cholesterol efflux assay, and expression of ABCA1 and cleaved caspase-3 were measured by western blotting. 10-14-week-old C57BL/6 male and female mice were given bilateral kidney X-irradiation of 4Gy with or without cisplatin (3mg/kg; IV). Functional kidney parameters, histopathological changes, and ultrastructural changes were measured at baseline and 20 weeks after treatment. GFR was measured using a FITC-sinistrin-based transdermal monitor. In vitro studies suggest that after radiation exposure (4Gy and 8Gy), ABCA1 expression decreases in a dose and time-dependent manner in cultured podocytes (p < 0.05), which coincides with reduced cholesterol efflux (p < 0.05) and increased podocyte apoptosis (p < 0.01). In contrast, LXR agonist treatment in vitro prevents reduced ABCA1 expression and cholesterol efflux and prevents podocytes from radiation-induced apoptosis. Our in vitro studies further suggest that Enzalutamide treatment alone reduces the growth of LNCaP and podocytes, and RT (5Gy) plus enzalutamide combination treatment further decreases the growth of LNCaP and podocytes. However, LXR agonist treatment further increased the efficacy of enzalutamide and RT treatment against LNCaP and increased podocyte growth simultaneously. Similarly, Sunitinib with or without RT reduces the growth of renal cancer cells (786-0) and podocytes. LXR agonist treatment further increases the efficacy of the sunitinib and RT against 786-0 cells and simultaneously increases podocyte growth. Cyclophosphamide reduces the growth of the podocytes, and LXR agonist treatment improves the podocyte growth after CP treatment. In vivo studies observed that focal bilateral kidney irradiation with or without cisplatin of C57BL/6 mice induces lipid accumulation in the kidney cortex and increases GBM thickness (p < 0.001), which correlates with decreased Abca1 expression, podocyte number, and GFR. This study shows that LXR agonist treatment protects podocytopathy in vitro and chemotherapy or radiotherapy-induced kidney injury in vivo. ABCA1 may be an important therapeutic target for chemoradiotherapy-induced kidney injuries in cancer patients.