Genomic structure of human lysosomal glycosylasparaginase

AbstractThe gene structure of the human lysosomal enzyme glycosylasparaginase was determined. The gene spans 13 kb and consists of 9 exons. Both 5′ and 3′ untranslated regions of the gene are uninterrupted by introns. A number of transcriptional elements were identified in the 5′ upstream sequence that includes two putative CAAT boxes followed by TATA-like sequences together with two AP-2 binding sites and one for Sp1. A 100 bp CpG island and several ETF binding sites were also found. Additional AP-2 and Sp1 binding sites are present in the first intron. Two polyadenylation sites are present and appear to be functional. The major known glycosylasparaginase gene defect G488→C, which causes the lysosomal storage disease aspartylglycosaminuria (AGU) in Finland, is located in exon 4. Exon 5 encodes the post-translational cleavage site for the formation of the mature α/β subunits of the enzyme as well as a recently proposed active site threonine, Thr206

Construction of a Stable Expression System Based on the Endogenous <i>hbpB</i>/<i>hbpC</i> Toxin–Antitoxin System of <i>Halomonas bluephagenesis</i>

Halomonas bluephagenesis is a halophilic bacterium capable of efficiently producing polyhydroxyalkanoates and other valuable chemicals through high salinity open fermentation, offering an appealing platform for next-generation industrial biotechnology. Various techniques have been developed to engineer Halomonas bluephagenesis, each with its inherent shortcomings. Genome editing methods often entail complex and time-consuming processes, while flexible expression systems relying on plasmids necessitate the use of antibiotics. In this study, we developed a stable recombinant plasmid vector, pHbPBC, based on a novel hbpB/hbpC toxin–antitoxin system found within the endogenous plasmid of Halomonas bluephagenesis. Remarkably, pHbPBC exhibited exceptional stability during 7 days of continuous subculture, eliminating the need for antibiotics or other selection pressures. This stability even rivaled genomic integration, all while achieving higher levels of heterologous expression. Our research introduces a novel approach for genetically modifying and harnessing nonmodel halophilic bacteria, contributing to the advancement of next-generation industrial biotechnology

Construction of a Stable Expression System Based on the Endogenous <i>hbpB</i>/<i>hbpC</i> Toxin–Antitoxin System of <i>Halomonas bluephagenesis</i>

Halomonas bluephagenesis is a halophilic bacterium capable of efficiently producing polyhydroxyalkanoates and other valuable chemicals through high salinity open fermentation, offering an appealing platform for next-generation industrial biotechnology. Various techniques have been developed to engineer Halomonas bluephagenesis, each with its inherent shortcomings. Genome editing methods often entail complex and time-consuming processes, while flexible expression systems relying on plasmids necessitate the use of antibiotics. In this study, we developed a stable recombinant plasmid vector, pHbPBC, based on a novel hbpB/hbpC toxin–antitoxin system found within the endogenous plasmid of Halomonas bluephagenesis. Remarkably, pHbPBC exhibited exceptional stability during 7 days of continuous subculture, eliminating the need for antibiotics or other selection pressures. This stability even rivaled genomic integration, all while achieving higher levels of heterologous expression. Our research introduces a novel approach for genetically modifying and harnessing nonmodel halophilic bacteria, contributing to the advancement of next-generation industrial biotechnology

TAK-242 suppresses cardiac inflammation and fibrosis caused by Aldo-salt <i>in vivo</i>.

<p>(A-C) Cardiac mRNA levels of TNF-α, MCP-1 and IL-1β in rats treated with Aldo-salt or with Aldo-salt plus TAK-242. *<i>p</i>< 0.05 vs control, #<i>p</i>< 0.05 vs Aldo-salt. (D) Cardiac protein levels of TNF-α, MCP-1 and IL-1β in rats treated with Aldo-salt or with Aldo-salt plus TAK-242 were assayed using ELISA. *<i>p</i>< 0.05 vs control, #<i>p</i>< 0.05 vs Aldo-salt.</p

Physiological and hematological parameters in Aldo-salt–treated rats.

<p>Aldo = aldosterone; SBP = systolic blood pressure; DBP = diastolic blood pressure; HW = heart weight; BW = body weight.</p><p>Values are presented as mean ± SEM.</p><p>*<i>p</i><0.05 vs. control</p><p>^#<i>p</i><0.05 vs. Aldo-salt group.</p><p>Physiological and hematological parameters in Aldo-salt–treated rats.</p

Cardiac and renal expression of TLR4 is increased in Aldo-salt-treated rats.

<p>(A and B) Rats were infused with Aldo-salt at 1 mg/kg/day for 4weeks, and cardiac mRNA levels (A) and protein levels (B) of TLR4 were assayed using qPCR and western blot, respectively. Rats were infused with Aldo-salt at 1 mg/kg/day for 4weeks, and renal mRNA levels (C) and protein levels (D) of TLR4 were assayed using qPCR and western blot, respectively. *<i>p</i>< 0.05.</p

All-Fabric Triboelectric Nanogenerator (AF-TENG) Smart Face Mask: Remote Long-Rate Breathing Monitoring and Apnea Alarm

Since the beginning of the COVID-19 pandemic, the use of face masks has become not only mandatory in several countries but also an acceptable approach for combating the pandemic. In the quest for designing an effective and useful face mask, triboelectric nanogenerators (TENGs) have been recently proposed. Novel functionalities are provided with the use of TENGs in face masks due to the induced triboelectrification generated by the exhaled and inhaled breath, allowing their use as an energy sensor. Nonetheless, within the face mask, the presence of nontextile plastics or other common triboelectric (TE) materials can be undesired. Herein, we propose the use of an all-fabric TENG (AF-TENG) with the use of high molecular weight polyethylene (UHMWPE) and cotton fabric as negative and positive triboelectric layers, respectively. With these materials, it is possible to detect the breathing of the patient, which in the case of not detecting a signal over a few minutes can trigger an alarm locally, providing valuable time. Also, in this article, we have sent breathing signals locally and remotely to distances up to 20 km via Wi-Fi and LoRa, the same as warning signals in the case of detecting anomalies. This work reveals the use of TENGs in smart face masks as an important tool to be used in difficult epidemiological periods to the general public, bringing much more comfort and relaxation to patients and elderly in today’s society, and based on pristine eco-friendly materials

All-Fabric Triboelectric Nanogenerator (AF-TENG) Smart Face Mask: Remote Long-Rate Breathing Monitoring and Apnea Alarm

Since the beginning of the COVID-19 pandemic, the use of face masks has become not only mandatory in several countries but also an acceptable approach for combating the pandemic. In the quest for designing an effective and useful face mask, triboelectric nanogenerators (TENGs) have been recently proposed. Novel functionalities are provided with the use of TENGs in face masks due to the induced triboelectrification generated by the exhaled and inhaled breath, allowing their use as an energy sensor. Nonetheless, within the face mask, the presence of nontextile plastics or other common triboelectric (TE) materials can be undesired. Herein, we propose the use of an all-fabric TENG (AF-TENG) with the use of high molecular weight polyethylene (UHMWPE) and cotton fabric as negative and positive triboelectric layers, respectively. With these materials, it is possible to detect the breathing of the patient, which in the case of not detecting a signal over a few minutes can trigger an alarm locally, providing valuable time. Also, in this article, we have sent breathing signals locally and remotely to distances up to 20 km via Wi-Fi and LoRa, the same as warning signals in the case of detecting anomalies. This work reveals the use of TENGs in smart face masks as an important tool to be used in difficult epidemiological periods to the general public, bringing much more comfort and relaxation to patients and elderly in today’s society, and based on pristine eco-friendly materials

Histological findings.

<p>Cardiac (A) or renal (B) mRNA levels of Col I and TGF-β in rats treated with Aldo-salt or with Aldo-salt plus TAK-242. *<i>p</i>< 0.05 vs control, #<i>p</i>< 0.05 vs Aldo-salt. (C)Representative images and quantitation of perivascular fibrosis in left ventricles of rats treated with Aldo-salt or with Aldo-salt plus TAK-242. *<i>p</i>< 0.05 vs control, #<i>p</i>< 0.05 vs Aldo-salt. (D) Histological staining with periodic acid Schiff (PAS), also showing tubulointerstitial damage of rats treated with Aldo-salt or with Aldo-salt plus TAK-242. *<i>p</i>< 0.05 vs control, #<i>p</i>< 0.05 vs Aldo-salt. n = 9, 200× magnification.</p

TAK-242 suppresses renal inflammation and fibrosis caused by Aldo-salt <i>in vivo</i>.

<p>(A-C) Renal mRNA levels of TNF-α, MCP-1 and IL-1β in rats treated with Aldo-salt or with Aldo-salt plus TAK-242. *<i>p</i>< 0.05 vs control, #<i>p</i>< 0.05 vs Aldo-salt. (D) Renal protein levels of TNF-α, MCP-1 and IL-1β in rats treated with Aldo-salt or with Aldo-salt plus TAK-242 were assayed using ELISA. *<i>p</i>< 0.05 vs control, #<i>p</i>< 0.05 vs Aldo-salt.</p