Engrailed: Pathological and physiological effects of a multifunctional developmental gene.

Engrailed-1 (EN1) is a developmental gene that encodes En1, a highly conserved transcription factor involved in regionalization during early embryogenesis and in the later maintenance of normal neurons. After birth, EN1 still plays a role in the development and physiology of the body; for example, it exerts a protective effect on midbrain dopaminergic (mDA) neurons, and loss of EN1 causes mDA neurons in the ventral midbrain to gradually die approximately 6 weeks after birth, resulting in motor and nonmotor symptoms similar to those observed in Parkinson's disease. Notably, EN1 has been identified as a possible susceptibility gene for idiopathic Parkinson's disease in humans. EN1 is involved in the processes of wound-healing scar production and tissue and organ fibrosis. Additionally, EN1 can lead to tumorigenesis and thus provides a target for the treatment of some tumors. In this review, we summarize the effects of EN1 on embryonic organ development, describe the consequences of the deletion or overexpression of the EN1 gene, and discuss the pathways in which EN1 is involved. We hope to clarify the role of EN1 as a developmental gene and present potential therapeutic targets for diseases involving the EN1 gene.

Diverse functions and pathogenetic role of Crumbs in retinopathy.

The Crumbs protein (CRB) family plays a crucial role in maintaining the apical-basal polarity and integrity of embryonic epithelia. The family comprises different isoforms in different animals and possesses diverse structural, localization, and functional characteristics. Mutations in the human CRB1 or CRB2 gene may lead to a broad spectrum of retinal dystrophies. Various CRB-associated experimental models have recently provided mechanistic insights into human CRB-associated retinopathies. The knowledge obtained from these models corroborates the importance of CRB in retinal development and maintenance. Therefore, complete elucidation of these models can provide excellent therapeutic prospects for human CRB-associated retinopathies. In this review, we summarize the current animal models and human-derived models of different CRB family members and describe the main characteristics of their retinal phenotypes.

The role of TET2-mediated ROBO4 hypomethylation in the development of diabetic retinopathy.

BACKGROUND: In diabetic retinopathy, increasing evidence points to a link between the pathogenesis of retinal microangiopathy and the endothelial cell-specific factor roundabout4 (ROBO4). According to earlier research, specificity protein 1 (SP1) enhances the binding to the ROBO4 promoter, increasing Robo4 expression and hastening the progression of diabetic retinopathy. To determine if this is related to aberrant epigenetic modifications of ROBO4, we examined the methylation level of the ROBO4 promoter and the corresponding regulatory mechanism during the course of diabetic retinopathy and explored the effect of this mechanism on retinal vascular leakage and neovascularization. METHODS: The methylation level of CpG sites in the ROBO4 promoter was detected in human retinal endothelial cells (HRECs) cultured under hyperglycemic conditions and retinas from streptozotocin-induced diabetic mice. The effects of hyperglycemia on DNA methyltransferase 1, Tet methylcytosine dioxygenase 2 (TET2), 5-methylcytosine, 5-hydroxymethylcytosine, and the binding of TET2 and SP1 to the ROBO4 promoter, as well as the expression of ROBO4, zonula occludens 1 (ZO-1) and occludin were examined. Short hairpin RNA was used to suppress the expression of TET2 or ROBO4 and the structural and functional changes in the retinal microvascular system were assessed. RESULTS: In HRECs cultured under hyperglycemic conditions, the ROBO4 promoter methylation level decreased. Hyperglycemia-induced TET2 overexpression caused active demethylation of ROBO4 by oxidizing 5-methylcytosine to 5-hydroxymethylcytosine, which enhanced the binding of SP1 to ROBO4, increased the expression of ROBO4, and decreased the expression of ZO-1 and occludin, leading to the abnormalities in monolayer permeability, migratory ability and angiogenesis of HRECs. The above pathway was also demonstrated in the retinas of diabetic mice, which caused leakage from retinal capillaries and neovascularization. Inhibition of TET2 or ROBO4 expression significantly ameliorated the dysfunction of HRECs and retinal vascular abnormalities. CONCLUSIONS: In diabetes, TET2 can regulate the expression of ROBO4 and its downstream proteins by mediating active demethylation of the ROBO4 promoter, which accelerates the development of retinal vasculopathy. These findings suggest that TET2-induced ROBO4 hypomethylation is a potential therapeutic target, and anti- TET2/ROBO4 therapy is anticipated to emerge as a novel strategy for early intervention and delayed progression of diabetic retinopathy.

Design, Synthesis, and Biological Evaluation of Benzimidazole Derivatives as Potential Lassa Virus Inhibitors.

The Lassa virus (LASV) causes Lassa fever, a highly infectious and lethal agent of acute viral hemorrhagic fever. At present, there are still no effective treatments available, creating an urgent need to develop novel therapeutics. Some benzimidazole compounds targeting the arenavirus envelope glycoprotein complex (GPC) are promising inhibitors of LASV. In this study, we synthesized two series of LASV inhibitors based on the benzimidazole structure. Lentiviral pseudotypes bearing the LASV GPC were established to identify virus entry inhibitors. Surface plasmon resonance (SPR) was further used to verify the binding activities of the potential compounds. Compounds 7d-Z, 7h-Z, 13c, 13d, and 13f showed relatively excellent antiviral activities with IC50 values ranging from 7.58 to 15.46 nM and their SI values above 1251. These five representative compounds exhibited stronger binding affinity with low equilibrium dissociation constants (KD < 8.25 × 10-7 M) in SPR study. The compound 7h-Z displayed the most potent antiviral activity (IC50 = 7.58 nM) with a relatively high SI value (2496), which could be further studied as a lead compound. The structure-activity relationship indicated that the compounds with lipophilic and spatially larger substituents might possess higher antiviral activity and a much larger safety margin. This study will provide some good guidance for the development of highly active compounds with a novel skeleton against LASV.

Mesoionic Carbene-Catalyzed Formyl Alkylation of Aldehydes.

Ketones are among the most useful functional groups in organic synthesis, and they are commonly encountered in a broad range of compounds with various applications. Herein, we describe the mesoionic carbene-catalyzed coupling reaction of aldehydes with non-activated secondary and even primary alkyl halides. This metal-free method utilizes deprotonated Breslow intermediates derived from mesoionic carbenes (MICs), which act as super electron donors and induce the single-electron reduction of alkyl halides. This mild coupling reaction has a broad substrate scope and tolerates many functional groups, which allows to prepare a diversity of simple ketones as well as bio-active molecules by late-stage functionalization.

Silica nanoparticle-modified microcomb electrode for voltammetry detection of osteopontin with high sensitivity.

Osteosarcoma is a commonly occurring bone malignancy, and it is the second most common cause of cancer deaths in adolescents and children. A sensitive silica nanoparticle (Si-NP) modified current-volt sensor was introduced to identify the osteopontin antigen, a well-known biomarker for osteosarcoma. Si-NP was extracted from the rice husk ash and utilized for the surface functionalization on the interdigitated microelectrode sensing surface. Extracted Si-NP has a spherical shape with uniform distribution, and it is confirmed by field emission scanning electron microscopy and field-emission transmission electron microscopy. Si-NP was layered on the electrode surface through a (3-aminopropyl)triethoxysilane amine linker, and the antibody was immobilized on Si-NP through a glutaraldehyde linker. Osteopontin was effectively detected on the antibody-attached surface, and the determination limit was 0.6 ng/mL. The regression was determined as y = 0.9366x - 1.1113 and the R2 value was 0.9331 and the detection limit of osteopontin was 0.6 ng/mL in the range between 0.3 and 5 ng/mL. In addition, control performance with nonimmune antibodies and albumin did not change the current volt, showing the specific osteopontin identification. This research work brings out the easy and cost-effective method to diagnose osteosarcoma and its etiology.

Base-Catalyzed H/D Exchange Reaction of Difluoromethylarenes.

The budding deuteriodifluoromethyl group (CF2D) is a potentially significant functional group in medicinal chemistry. Herein, we investigated t-BuOK-catalyzed H/D exchange reaction of difluoromethylarenes in DMSO-d6 solution. The method provides excellent deuterium incorporation at the difluoromethyl group. Meanwhile, the effect of a trace amount of D2O in DMSO-d6 solution on the deuteration reaction was also investigated.

The effect of diabetes on corneal endothelium: a meta-analysis.

BACKGROUND: This research was conducted with the aim to determine the effect of diabetes mellitus on corneal endothelial cells. METHODS: The terms: ("diabetes mellitus" or "diabetes" or "diabetic") and ("corneal endothelium" or "cornea" or "Corneas") searched in Pubmed, Embase, Cochrane, and Web of science until August 2019. The included types of studies contained observational studies. The standard mean difference (SMD) which was deemed as main size effects for continuous data was calculated by means and standard deviations. The data on corneal endothelial cell density (ECD), mean cell area (MCA), cell area variation coefficient (CV) and percentage of hexagonal cells (HEX) included in the study were collected and analyzed using stata15.1. RESULTS: The final 16 cross-sectional studies and 2 case-control studies were included for the meta-analysis. Meta-analysis revealed that diabetes mellitus could reduce ECD (SMD = - 0.352, 95% CI -0.538, - 0.166) and the HEX (SMD = - 0.145, 95% CI -0.217, - 0.074), in addition to increasing CV (SMD = 0.195, 95% CI 0.123, 0.268). Nevertheless, there was no statistically significant differences observed when combining MCA (SMD = 0.078, 95% CI -0.022, 0.178). In subgroup analysis, Type 2 diabetes patients owned less corneal ECD (P < 0.05). Moreover the same results also found during the subgroup form Asia, Europe and American. The meta-regression revealed the type of diabetes mellitus might be contributing to heterogeneity. (P = 0.008). The results indicated a significant publication bias for studies, with combined CV (Begg's test, P = 0.006; Egger's test, P = 0.005) and merged combined HEX (Begg's test, P = 0.113; Egger's test, P = 0.024). CONCLUSIONS: As indicated by meta-analysis, diabetes mellitus could cause a detrimental effect on corneal endothelium health. Diabetes mellitus contributed to the instability of corneal endothelium during the analysis. Therefore, further research is considered necessary to confirm our research results. TRIAL REGISTRATION: CED 42019145858 .

Synthesis, Structure, and Photophysical Properties of m-Phenylene-Embedded Cycloparaphenylene Nanorings.

Five m-phenylene-embedded cycloparaphenylenes m3[9]CPP 1-5 were synthesized by the platinum-mediated cyclooligomerization strategy with high overall yields. The structures of m3[9]CPP 1-3 were determined by X-ray diffraction analysis. Compared to [9]CPP, m3[9]CPP 1 caused a significant blueshift in the UV-vis absorption and fluorescence spectra. This result shows that the radial π-conjugation is distorted and partially interrupted. The photophysical properties of m3[9]CPP 1 were further tuned by the introduction of various substituents for m3[9]CPP 2-5. Methoxy group substitution at m-phenylene did not change the photophysical properties significantly. Replacement of m-phenylene by tetrafluoro-m-phenylene achieved a significant blueshift. When the carboxyl group was embedded at m-phenylene or the methoxy group was embedded at p-phenylene, significant redshifts were observed with blue color emission. Theoretical calculations revealed that the decrease in the HOMO-LUMO gap in m3[9]CPP 4 and 5 is favorable for the redshift of the fluorescence spectrum.

Iron-induced Local Complement Component 3 (C3) Up-regulation via Non-canonical Transforming Growth Factor (TGF)-ß Signaling in the Retinal Pigment Epithelium.

Dysregulation of iron homeostasis may be a pathogenic factor in age-related macular degeneration (AMD). Meanwhile, the formation of complement-containing deposits under the retinal pigment epithelial (RPE) cell layer is a pathognomonic feature of AMD. In this study, we investigated the molecular mechanisms by which complement component 3 (C3), a central protein in the complement cascade, is up-regulated by iron in RPE cells. Modulation of TGF-ß signaling, involving ERK1/2, SMAD3, and CCAAT/enhancer-binding protein-δ, is responsible for iron-induced C3 expression. The differential effects of spatially distinct SMAD3 phosphorylation sites at the linker region and at the C terminus determined the up-regulation of C3. Pharmacologic inhibition of either ERK1/2 or SMAD3 phosphorylation decreased iron-induced C3 expression levels. Knockdown of SMAD3 blocked the iron-induced up-regulation and nuclear accumulation of CCAAT/enhancer-binding protein-δ, a transcription factor that has been shown previously to bind the basic leucine zipper 1 domain in the C3 promoter. We show herein that mutation of this domain reduced iron-induced C3 promoter activity. In vivo studies support our in vitro finding of iron-induced C3 up-regulation. Mice with a mosaic pattern of RPE-specific iron overload demonstrated co-localization of iron-induced ferritin and C3d deposits. Humans with aceruloplasminemia causing RPE iron overload had increased RPE C3d deposition. The molecular events in the iron-C3 pathway represent therapeutic targets for AMD or other diseases exacerbated by iron-induced local complement dysregulation.

Contributions of phosphatidylserine-positive platelets and leukocytes and microparticles to hypercoagulable state in gastric cancer patients.

Hypercoagulability in gastric cancer is a common complication and a major contributor to poor prognosis. This study aimed to determine procoagulant activity of blood cells and microparticles (MPs) in gastric cancer patients. Phosphatidylserine-positive blood cells and MPs, and their procoagulant properties in particular, were assessed in 48 gastric cancer patients and 35 healthy controls. Phosphatidylserine-positive platelets, leukocytes, and MPs in patients with tumor-node-metastasis stage III/IV gastric cancer were significantly higher than those in stage I/II patients or healthy controls. Moreover, procoagulant activity of platelets, leukocytes, and MPs in stage III/IV patients was significantly increased compared to the controls, as indicated by shorter clotting time, higher intrinsic and extrinsic factor tenase, and prothrombinase complex activity. Interestingly, lactadherin, which competes with factors V and VIII to bind phosphatidylserine, dramatically prolonged clotting time of the cells and MPs by inhibiting factor tenase and prothrombinase complex activity. Although anti-tissue factor antibody significantly attenuated extrinsic tenase complex activity of leukocytes and MPs, it only slightly prolonged clotting times. Meanwhile, treatment with radical resection reduced phosphatidylserine-positive platelets, leukocytes, and MPs, and prolonged the clotting times of the remaining cells and MPs. Our results suggest that phosphatidylserine-positive platelets, leukocytes, and MPs contribute to hypercoagulability and represent a potential therapeutic target to prevent coagulation in patients with stage III/IV gastric cancer.

Cp/Heph mutant mice have iron-induced neurodegeneration diminished by deferiprone.

Brain iron accumulates in several neurodegenerative diseases and can cause oxidative damage, but mechanisms of brain iron homeostasis are incompletely understood. Patients with mutations in the cellular iron-exporting ferroxidase ceruloplasmin (Cp) have brain iron accumulation causing neurodegeneration. Here, we assessed the brains of mice with combined mutation of Cp and its homolog hephaestin. Compared to single mutants, brain iron accumulation was accelerated in double mutants in the cerebellum, substantia nigra, and hippocampus. Iron accumulated within glia, while neurons were iron deficient. There was loss of both neurons and glia. Mice developed ataxia and tremor, and most died by 9 months. Treatment with the oral iron chelator deferiprone diminished brain iron levels, protected against neuron loss, and extended lifespan. Ferroxidases play important, partially overlapping roles in brain iron homeostasis by facilitating iron export from glia, making iron available to neurons. Above: Iron (Fe) normally moves from capillaries to glia to neurons. It is exported from the glia by ferroportin (Fpn) with ferroxidases ceruloplasmin (Cp) and/or Hephaestin (Heph). Below: In mice with mutation of Cp and Heph, iron accumulates in glia, while neurons have low iron levels. Both neurons and glia degenerate and mice become ataxic unless given an iron chelator.

A high serum iron level causes mouse retinal iron accumulation despite an intact blood-retinal barrier.

The retina can be shielded by the blood-retinal barrier. Because photoreceptors are damaged by excess iron, it is important to understand whether the blood-retinal barrier protects against high serum iron levels. Bone morphogenic protein 6 (Bmp6) knockout mice have serum iron overload. Herein, we tested whether the previously documented retinal iron accumulation in Bmp6 knockout mice might result from the high serum iron levels or, alternatively, low levels of retinal hepcidin, an iron regulatory hormone whose transcription can be up-regulated by Bmp6. Furthermore, to determine whether increases in serum iron can elevate retinal iron levels, we i.v. injected iron into wild-type mice. Retinas were analyzed by real-time quantitative PCR and immunofluorescence to assess the levels of iron-regulated genes/proteins and oxidative stress. Retinal hepcidin mRNA levels in Bmp6 knockout retinas were the same as, or greater than, those in age-matched wild-type retinas, indicating that Bmp6 knockout does not cause retinal hepcidin deficiency. Changes in mRNA levels of L ferritin and transferrin receptor indicated increased retinal iron levels in i.v. iron-injected wild-type mice. Oxidative stress markers were elevated in photoreceptors of mice receiving i.v. iron. These findings suggest that elevated serum iron levels can overwhelm local retinal iron regulatory mechanisms.

A murine RP1 missense mutation causes protein mislocalization and slowly progressive photoreceptor degeneration.

Mutations in the RP1 gene can cause retinitis pigmentosa. We identified a spontaneous L66P mutation caused by two adjacent point mutations in the Rp1 gene in a colony of C57BL/6J mice. Mice homozygous for the L66P mutation exhibited slow, progressive photoreceptor degeneration throughout their lifespan. Optical coherence tomography imaging found abnormal photoreceptor reflectivity at 1 month of age. Histology found shortening and disorganization of the photoreceptor inner and outer segments and progressive thinning of the outer nuclear layer. Electroretinogram a- and b-wave amplitudes were decreased with age. Western blot analysis found that the quantity and size of the mutated retinitis pigmentosa 1 (RP1) protein were normal. However, immunohistochemistry found that the mutant Rp1 protein partially mislocalized to the transition zone of the shortened axonemes. This mutation disrupted colocalization with cytoplasmic microtubules in vitro. In conclusion, the L66P mutation in the first doublecortin domain of the Rp1 gene impairs Rp1 protein localization and function, leading to abnormalities in photoreceptor outer segment structure and progressive photoreceptor degeneration. This is the first missense mutation in Rp1 shown to cause retinal degeneration. It provides a unique, slowly progressive photoreceptor degeneration model that mirrors the slow degeneration kinetics in most patients with retinitis pigmentosa.

In vitro interaction of naphthoquine with ivermectin, atovaquone, curcumin, and ketotifen in the asexual blood stage of Plasmodium falciparum 3D7.

Naphthoquine is a promising candidate for antimalarial combination therapy. Its combination with artemisinin has demonstrated excellent efficacy in clinical trials conducted across various malaria-endemic areas. A co-formulated combination of naphthoquine and azithromycin has also shown high clinical efficacy for malaria prophylaxis in Southeast Asia. Developing new combination therapies using naphthoquine will provide additional arsenal responses to the growing threat of artemisinin resistance. Furthermore, due to its long half-life, the possible interaction of naphthoquine with other drugs also needs attention. However, studies on its pharmacodynamic interactions with other drugs are still limited. In this study, the in vitro interactions of naphthoquine with ivermectin, atovaquone, curcumin, and ketotifen were evaluated in the asexual stage of Plasmodium falciparum 3D7. By using the combination index analysis and the SYBR Green I-based fluorescence assay, different interaction patterns of selected drugs with naphthoquine were revealed. Curcumin showed a slight but significant synergistic interaction with naphthoquine at lower effect levels, and no antagonism was observed across the full range of effect levels for all tested ratios. Atovaquone showed a potency decline when combined with naphthoquine. For ivermectin, a significant antagonism with naphthoquine was observed at a broad range of effect levels below 75% inhibition, although no significant interaction was observed at higher effect levels. Ketotifen interacted with naphthoquine similar to ivermectin, but significant antagonism was observed for only one tested ratio. These findings should be helpful to the development of new naphthoquine-based combination therapy and the clinically reasonable application of naphthoquine-containing therapies. IMPORTANCE: Pharmacodynamic interaction between antimalarials is not only crucial for the development of new antimalarial combination therapies but also important for the appropriate clinical use of antimalarials. The significant synergism between curcumin and naphthoquine observed in this study suggests the potential value for further development of new antimalarial combination therapy. The finding of a decline in atovaquone potency in the presence of naphthoquine alerts to a possible risk of treatment or prophylaxis failure for atovaquone-proguanil following naphthoquine-containing therapies. The observation of antagonism between naphthoquine and ivermectin raised a need for concern about the applicability of naphthoquine-containing therapy in malaria-endemic areas with ivermectin mass drug administration deployed. Considering the role of atovaquone-proguanil as a major alternative when first-line artemisinin-based combination therapy is ineffective and the wide implementation of ivermectin mass drug administration in malaria-endemic countries, the above findings will be important for the appropriate clinical application of antimalarials involving naphthoquine-containing therapies.

[The prevalence and influencing factors of eye diseases for IT industry video operation workers].

OBJECTIVE: To investigate the situation of video-contact and eye diseases for IT industry video operation workers, and to analyze the influencing factors, providing scientific evidence for the make of health-strategy for IT industry video operation workers. METHODS: We take the random cluster sampling method to choose 190 IT industry video operation workers in a city of Jiangsu province, analyzing the relations between video contact and eye diseases. RESULTS: The daily video contact time of IT industry video operation workers is 6.0-16.0 hours, whose mean value is (I 0.1 ± 1.8) hours. 79.5% of workers in this survey wear myopic lens, 35.8% of workers have a rest during their working, and 14.2% of IT workers use protective products when they feel unwell of their eyes. Following the BUT experiment, 54.7% of IT workers have the normal examine results of hinoculus, while 45.3% have the abnormal results of at least one eye. Simultaneously, 54.7% workers have the normal examine results of hinoculus in the SIT experiment, however, 42.1% workers are abnormal. According to the broad linear model, there are six influencing factors (daily mean time to video, distance between eye and displayer, the frequency of rest, whether to use protective products when they feel unwell of their eyes, the type of dis player and daily time watching TV.) have significant influence on vision, having statistical significance. At the same time, there are also six influencing factors (whether have a rest regularly,sex, the situation of diaphaneity for cornea, the shape of pupil, family history and whether to use protective products when they feel unwell of their eyes.) have significant influence on the results of BUT experiment,having statistical significance. However, there are seven influencing factors (the type of computer, sex, the shape of pupil, the situation of diaphaneity for cornea, the angle between displayer and workers' sight, the type of displayer and the height of operating floor.) have significant influence on the results of SIT experiment,having statistical significance. CONCLUSIONS: The health-situation of IT industry video operation workers' eye is not optimistic, most of workers are lack of protection awareness; we need to strengthen propaganda and education according to its influencing factors and to improve the level of medical control and prevention for eye diseases in relevant industries.

Cutting-Edge Research in Sports Biomechanics: From Basic Science to Applied Technology.

Sports biomechanics is the study of the mechanical principles of human movement and how they apply to sports performance [...].

Evaluation of the Effect of Preharvest Melatonin Spraying on Fruit Quality of 'Yuluxiang' Pear Based on Principal Component Analysis.

Melatonin (MT), an indoleamine compound, has a pleiotropic effect on plant growth and development and can regulate the quality of tree fruit. Systematic research on the effect of preharvest MT spraying on pear fruit quality and technical solutions for MT application to regulate pear fruit quality are still lacking. Thus, here we aimed to evaluate the effects of different spraying times, concentrations, and exogenous MT application times on 'Yuluxiang' pear fruit quality. Our results showed that the single fruit weight and vertical and horizontal diameters of pear fruit sprayed with MT twice at 30 and 90 d after full bloom were the largest, and the red and green values of the treatment were the highest. MT-treated pears had higher contents of total soluble solids, soluble sugar, sucrose, sorbitol, fructose, and glucose and lower contents of titratable acid, malic acid, and citric acid. Moreover, exogenous MT treatment increased the pear peel strength. Based on the principal component analysis of 10 fruit quality indices, the suitable periods for MT spraying on 'Yuluxiang' pears were 30 and 90 d after full bloom, the suitable concentration was 100 µmol/L, and the suitable number of times was two. This study provides a theoretical reference for optimizing MT application and improving pear fruit quality.

Conducting quantitative mask fit tests: application details and affecting factors.

Introduction: Respirators chosen based on their assigned protection factor (APF) enable wearers to effectively reduce particulate matter concentrations to safe levels when used correctly. As a crucial factor in achieving the intended APF, the fit test has become a necessary procedure in respiratory disease protection. Methods: This study involved 225 participants who underwent a fit test using two reusable types of half masks and two types of full masks. Condensation nuclei counting (CNC) and controlled negative pressure (CNP) were performed. Results: The results revealed that the passing rate of full masks was higher compared to half masks. Specifically, the passing rate for the half masks and the full masks were 84.7 and 91.6%, respectively. Gender exerted a statistically significant effect on the passing rate. Nevertheless, age, educational background, and training exhibited relatively negligible effects. Certain movements, such as facing forward, were identified as key actions with strong correlation. Additionally, talking was considered a key action with a high failure rate due to instantaneous leakages. Most participants failed at the initial step of CNP, but nearly all of them passed the fit test using CNC. Discussion: Therefore, putting on full masks, especially for women, provides optimal protection during work. Furthermore, attention should be given to the displacement and deformation of the respirator during the key actions. When it comes to fit test methods, CNC was found to be more practical and comprehensive compared to CNP. Moreover, additional physiological characteristics, such as double chins, could be explored as potential influential factors.

Disulfide-incorporated lipid prodrugs of cidofovir: Synthesis, antiviral activity, and release mechanism.

The double-stranded DNA (dsDNA) viruses represented by adenovirus and monkeypox virus, have attracted widespread attention due to their high infectivity. In 2022, the global outbreak of mpox (or monkeypox) has led to the declaration of a Public Health Emergency of International Concern. However, to date therapeutics approved for dsDNA virus infections remain limited and there are still no available treatments for some of these diseases. The development of new therapies for treating dsDNA infection is in urgent need. In this study, we designed and synthesized a series of novel disulfide-incorporated lipid conjugates of cidofovir (CDV) as potential candidates against dsDNA viruses including vaccinia virus (VACV) and adenovirus (AdV) 5. The structure-activity relationship analyses revealed that the optimum linker moiety was C2H4 and the optimum aliphatic chain length was 18 or 20 atoms. Among the synthesized conjugates, 1c exhibited more potency against VACV (IC50 = 0.0960 µM in Vero cells; IC50 = 0.0790 µM in A549 cells) and AdV5 (IC50 = 0.1572 µM in A549 cells) than brincidofovir (BCV). The transmission electron microscopy (TEM) images revealed that the conjugates could form micelles in phosphate buffer. The stability studies in the GSH environment demonstrated that the formation of micelles in phosphate buffer might protect the disulfide bond from glutathione (GSH) reduction. The dominant means of the synthetic conjugates to liberate the parent drug CDV was by enzymatic hydrolysis. Furthermore, the synthetic conjugates remained sufficiently stable in simulated gastric fluid (SGF), simulated intestinal fluid (SIF), and pooled human plasma, which indicated the possibility for oral administration. These results indicated 1c may be a broad-spectrum antiviral candidate against dsDNA viruses with potential oral administration. Moreover, modification of the aliphatic chain attached to the nucleoside phosphonate group was involved as an efficient prodrug strategy for the development of potent antiviral candidates.