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Neuroinflammatory conditions linked to iron dysregulation pose significant challenges in neurodegenerative diseases. Iron‐loaded microglia are observed in the brains of patients with various neuroinflammatory conditions, yet how iron overload affects microglial function and contributes to various neuroinflammatory processes is poorly understood. This in vitro study elucidates the relationship between excess iron, cofilin activation, and microglial function, shedding light on potential therapeutic avenues. Iron overload was induced in Human Microglial Clone 3 cells using ferrous sulfate, and the expressions of ferritin heavy chain, ferritin light chain, divalent metal transporter 1, cofilin, p‐cofilin, nuclear factor‐κB (NF‐κB), and various inflammatory cytokines were analyzed using real‐time quantitative polymerase chain reaction, immunocytochemistry, Western blotting, and enzyme‐linked immunosorbent assay. Results revealed a notable increase in cofilin, NF‐κB, and inflammatory cytokine expression levels following excess iron exposure. Moreover, treatment with deferoxamine (DFX), a known iron chelator, and a novel cofilin inhibitor (CI) synthesized in our laboratory demonstrate a mitigating effect on iron‐induced cofilin expression. Furthermore, both DFX and CI exhibit promising outcomes in mitigating the inflammatory consequences of excess iron, including the expression of pro‐inflammatory cytokines and NF‐κB activation. These findings suggest that both DFX and CI can potentially alleviate microglia‐induced neuroinflammation by targeting both iron dysregulation and cofilin‐mediated pathways. Overall, this study provides valuable insights into iron‐induced cofilin activation and microglial activation, offering avenues for potential targeted therapies for neuroinflammatory conditions associated with iron and cofilin dysregulation in neurodegenerative diseases. [ABSTRACT FROM AUTHOR]

A major breakthrough in developmental biology is the ex vivo generation of synthetic human embryos from stem cells. A comprehensive, in-depth bioethical analysis from a Sunni Islamic perspective reveals that the reproductive applications of synthetic human embryos contravene Islamic precepts of preserving lineage integrity (Hifz al-Nasl) due to disruption and confusion of kinship and familial relationships, similar to human cloning with somatic cell nuclear transfer. However, their non-reproductive applications in generating replacement tissues/organs, serving as in vitro experimental models of human development and disease, and testing platforms for evaluating pharmaceuticals and biomedical devices appear to align with Islamic principles. [ABSTRACT FROM AUTHOR]

Apolipoprotein-L1 (APOL1) is a membrane-interacting protein induced by inflammation, which confers human resistance to infection by African trypanosomes. APOL1 kills Trypanosoma brucei through induction of apoptotic-like parasite death, but two T. brucei clones acquired resistance to APOL1, allowing them to cause sleeping sickness. An APOL1 C-terminal sequence alteration, such as occurs in natural West African variants G1 and G2, restored human resistance to these clones. However, APOL1 unfolding induced by G1 or G2 mutations enhances protein hydrophobicity, resulting in kidney podocyte dysfunctions affecting renal filtration. The mechanism involved in these dysfunctions is debated. The ability of APOL1 to generate ion pores in trypanosome intracellular membranes or in synthetic membranes was provided as an explanation. However, transmembrane insertion of APOL1 strictly depends on acidic conditions, and podocyte cytopathology mainly results from secreted APOL1 activity on the plasma membrane, which occurs under non-acidic conditions. In this review, I argue that besides inactivation of APOL3 functions in membrane dynamics (fission and fusion), APOL1 variants induce inflammation-linked podocyte toxicity not through pore formation, but through plasma membrane disturbance resulting from increased interaction with cholesterol, which enhances cation channels activity. A natural mutation in the membrane-interacting domain (N264K) abrogates variant APOL1 toxicity at the expense of slightly increased sensitivity to trypanosomes, further illustrating the continuous mutual adaptation between host and parasite. [ABSTRACT FROM AUTHOR]

Microglia are resident brain cells that regulate neuronal development and innate immunity. Microglia activation participates in the cellular response to neuroinflammation, thus representing a possible target for pharmacological strategies aimed to counteract the onset and progression of brain disorders, including depression. Antidepressant drugs have been reported to reduce neuroinflammation by acting also on glial cells. Herein, the potential anti‐inflammatory and neuroprotective effects of trazodone (TRZ) on the microglial human microglial clone 3 (HMC3) cell line were investigated. HMC3 cells were activated by a double inflammatory stimulus (lipopolysaccharide [LPS] and tumour necrosis factor‐alpha [TNF‐α], 24 h each), and the induction of inflammation was demonstrated by (i) the increased expression levels of nuclear factor kappa‐light‐chain‐enhancer of activated B cells (NF‐kB) and ionized calcium‐binding adapter molecule 1 (IBA‐1), and (ii) the increased release of interleukin 6 (IL‐6) and transforming growth factor‐beta (TGF‐β). TRZ effects were evaluated by treating HMC3 cells for 24 h before (pre‐treatment) and after (post‐treatment) the double inflammatory stimulus. Notably, TRZ treatments significantly decreased the expression of NF‐kB and IBA‐1 and the release of the cytokines IL‐6 and TGF‐β. Moreover, TRZ prevented and reduced the release of quinolinic acid (QUIN), a known neurotoxic kynurenine metabolite. Finally, cellular supernatants collected from microglial cells pre‐treated LPS‐TNF‐α with TRZ were able to improve neuronal‐like cell viability, demonstrating a potential neuroprotective effect. Overall, this study suggests the anti‐inflammatory effects of TRZ on human microglia and strives for its neuroprotective properties. [ABSTRACT FROM AUTHOR]

Microglia serve as the primary defense mechanism in the brain. Artificial sweeteners are widely used as dietary supplements, though their long‐term effects remain uncertain. In this study, we investigated the effects of sucralose on microglia during prolonged exposure via the neuroinflammatory and ferroptosis pathways. Initially, human microglial clone 3 (HMC3) cells were exposed to sucralose (0–50 mM) for 24, 48, and 72 h to investigate the short‐term effects. Subsequently, HMC3 cells were treated with 1 mM sucralose for 7, 14, and 21 days to examine long‐term effects. We measured levels of interleukin‐1β (IL‐1β), NOD‐like receptor protein 3 (NLRP3), 8‐hydroxydeoxyguanosine (8‐OHdG), Sirtuin‐1 (SIRT1), glutathione peroxidase‐4 (GPx4), reduced glutathione (GSH), malondialdehyde (MDA), ferrous iron (Fe2+), and caspase 3/7. Additionally, we analyzed the impact of sucralose on cell morphology, migration, and expression levels of IL‐1β, NLRP3, SIRT1, and GPx4. Sucralose inhibited cell viability and proliferation in HMC3 cells in a concentration‐ and time‐dependent manner and induced membrane and nuclear abnormalities. Moreover, sucralose significantly reduced the cell migration rate. Long‐term sucralose treatment decreased Fe2+, GPx4, GSH, and SIRT1 levels in HMC3 cells while increasing IL‐1β, MDA, NLRP3, 8‐OHdG, and caspase 3/7 activity. Sucralose treatment also enhanced microglial activation and neuroinflammation by upregulating IL‐1β and NLRP3 and downregulating SIRT1 and GPx4, thereby inducing ferroptosis and suppressing cell viability. Consequently, high concentrations or long‐term sucralose treatment may induce neuroinflammation and ferroptosis by targeting the SIRT1/NLRP3/IL‐1β/GPx4 pathway in HMC3 cells. [ABSTRACT FROM AUTHOR]

The article highlights several papers about human life that were previously published in the periodical. Topics mentioned include the role of journalists in restoring justice to unborn children, a proposal for a constitutional amendment concerning anti-abortion law and the issues concerning the protection of human life.

One of the methods of cancer treatment is radiotherapy. Although radiation resistance and toxicity in normal cells limit the use of radiotherapy in some specific anatomical situations, however, together with radiotherapy, the use of substances that increase the radiation sensitivity of cancer cells and at the same time toxicity can be useful if they do not have cells on normal cells. The present study aimed to investigate the radiosensitizing effect of tolmetin in radiotherapy treatment on human clonal cancer cells. In this study, human clone HT-29 cancer cells in different groups were exposed to X-rays and tolmetin drug and were compared with each other and with the control group by micronucleus evaluation method and nuclear division index. The effect of cytotoxicity was evaluated with the index of nuclear division and genotoxicity of cells with the method of evaluating the number of micronuclei. Based on the results, the number of micronuclei increased significantly in the radiation-receiving group compared to the control group. In the group receiving tolmetin with 75 and 100 μM concentrations, the number of micronuclei also increased compared to the control group. A significant increase in the number of micronuclei was observed in all groups treated with tolmetin that received radiation, and this increase was more noticeable in concentrations of 100 and 150 μM. Meanwhile, tolmetin did not change the nuclear division index in the studied concentrations. The present study showed that tolmetin has a radiosensitizing effect on HT-29 human colon cancer cells, which is dependent on tolmetin concentration. In addition, tolmetin does not have a cytotoxicity effect on the mentioned cell line. [ABSTRACT FROM AUTHOR]

In recent years, some scientists have called for research into and potential development of 'solar geoengineering' technologies as an option to counter global warming. Solar geoengineering refers to a set of speculative techniques to reflect some incoming sunlight back into space, for example, by continuously spraying reflective sulphur aerosols into the stratosphere over several generations. Because of the significant ecological, social, and political risks posed by such technologies, many scholars and civil society organizations have urged governments to take action to prohibit the development and deployment of solar geoengineering techniques. In this article we take such calls for a prohibitory or a non-use regime on solar geoengineering as a starting point to examine existing international law and governance precedents that could guide the development of such a regime. The precedents we examine include international prohibitory and restrictive regimes that impose bans or restrictions on chemical weapons, biological weapons, weather modification technologies, anti-personnel landmines, substances that deplete the ozone layer, trade in hazardous wastes, deep seabed mining, and mining in Antarctica. We also assess emerging norms and soft law in anticipatory governance of novel technologies, such as human cloning and gene editing. While there is no blueprint for a solar geoengineering non-use regime in international law, our analysis points to numerous specific elements on which governments could draw to constrain or impose an outright prohibition on the development of technologies for solar geoengineering, should they opt to do so. [ABSTRACT FROM AUTHOR]

Copyright of Brazilian Journal of Law & International Relations / Relações Internacionais no Mundo is the property of Relacoes Internacionais no Mundo and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Filosofija, Sociologija is the property of Lithuanian Academy of Sciences Publishers and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

The article published in the British Journal of Dermatology discusses the impact of photodynamic therapy (PDT) on reducing clonal mutations in human and mouse skin caused by ultraviolet exposure. The study analyzed skin samples from patients undergoing PDT for actinic keratoses (AKs) and found a significant reduction in clonal mutations, particularly low variant allele frequency (VAF) mutations. The research also included a mouse model, showing similar results with fewer low VAF mutations in PDT-treated skin samples compared to controls. The study suggests that low VAF clonal mutations could be a potential biomarker for evaluating treatment efficacy. [Extracted from the article]

Hybrid insulin peptides (HIPs) formed through covalent cross-linking of proinsulin fragments to secretory granule peptides are detectable within murine and human islets. The 2.5HIP (C-peptide–chromogranin A [CgA] HIP), recognized by the diabetogenic BDC-2.5 clone, is a major autoantigen in the nonobese diabetic mouse. However, the relevance of this epitope in human disease is currently unclear. A recent study probed T-cell reactivity toward HIPs in patients with type 1 diabetes, documenting responses in one-third of the patients and isolating several HIP-reactive T-cell clones. In this study, we isolated a novel T-cell clone and showed that it responds vigorously to the human equivalent of the 2.5HIP (designated HIP9). Although the responding patient carried the risk-associated DRB1*04:01/DQ8 haplotype, the response was restricted by DRB1*11:03 (DR11). HLA class II tetramer staining revealed higher frequencies of HIP9-reactive T cells in individuals with diabetes than in control participants. Furthermore, in DR11+ participants carrying the DRB4 allele, HIP9-reactive T-cell frequencies were higher than observed frequencies for the immunodominant proinsulin 9-28 epitope. Finally, there was a negative correlation between HIP9-reactive T-cell frequency and age at diagnosis. These results provide direct evidence that this C-peptide–CgA HIP is relevant in human type 1 diabetes and suggest a mechanism by which nonrisk HLA haplotypes may contribute to the development of β-cell autoimmunity. Article Highlights: A hybrid insulin peptide formed from C-peptide–WE14 is a potent ligand for the diabetogenic BDC-2.5 clone, but its relevance in human disease is unclear. A human T-cell clone specific for the C-peptide–WE14 HIP is restricted by HLA-DRB1*11, a nonrisk allele. Using an HLA class II tetramer, we observed increased frequencies of HIP-reactive cells T cells in individuals with diabetes compared with control participants and an inverse correlation between frequency and age of onset. These results suggest a pathogenic role for the C-peptide–WE14 HIP and provide a plausible mechanism for the expansion of disease-driving T-cell responses in individuals with nonrisk HLA haplotypes. [ABSTRACT FROM AUTHOR]

Postoperative cognitive dysfunction is a common complication following anesthesia and surgery, and is associated with oxidative stress and apoptosis in the central nervous system. Resveratrol, an antioxidant, natural polyphenol, has been reported to have neuroprotective effect. The aim of this study was to examine whether resveratrol could attenuate the apoptosis and oxidative stress in a cell model of postoperative cognitive dysfunction. A cell model of postoperative cognitive dysfunction was constructed using human microglial clone 3 cells exposed to 1000 ng/ml lipopolysaccharide, 50 µM resveratrol, and their combination for 48 h. Then, the cell viability, proliferation, apoptosis, production of reactive oxygen species, and the expression of apoptosis-related genes and nuclear factor-E2-related factor 2 and its downstream antioxidant genes were examined. In the postoperative cognitive dysfunction cell model, lipopolysaccharide decreased the cell viability and proliferation, while it increased cell apoptosis and reactive oxygen species level. Lipopolysaccharide also upregulated the expression of pro-apoptotic genes (Bcl-2-associated X-protein and caspase-3) and genes involved in nuclear factor-E2-related factor 2-mediated antioxidant pathway (nuclear factor-E2-related factor 2, Kelch-like ECH-associated protein 1, heme oxygenase-1, and superoxide dismutase-2) in human microglial clone 3 cells. Resveratrol ameliorated the lipopolysaccharide-induced decrease of cell viability and proliferation, increase of cell apoptosis and reactive oxygen species level, and upregulation of the expression of pro-apoptotic genes. Additionally, resveratrol upregulated the expression of nuclear factor-E2-related factor 2 and its downstream antioxidant genes in lipopolysaccharide-treated human microglial clone 3 cells. Resveratrol protects human microglial clone 3 cells against lipopolysaccharide-induced injury by reducing oxidative stress through activating nuclear factor-E2-related factor 2-mediated antioxidant pathway. [ABSTRACT FROM AUTHOR]

Nancy Farmer's novel The House of the Scorpion deals with complex ethical dilemmas stemming from scientific advancements, particularly in cloning. The story unfolds in a dystopian society where clones are treated as subhuman entities. They are raised solely for the purpose of being harvested for their organs. This societal structure starkly contrasts the ethical and moral dilemmas associated with cloning. The paper highlights the stark contrast between the protected bodies of the ruling class and the exploited bodies of the clones and eejits, exposing the evils of totalitarianism and the misuse of technology. This analysis employs the ethical concepts organized in the book, The Ethics of Human Cloning, to explore the ethical dimensions surrounding human cloning. The study is targeted at stressing the necessity for implementing stringent ethical guidelines. [ABSTRACT FROM AUTHOR]

Treating hemophilia A patients who develop inhibitors remains a clinical challenge. A mouse model of hemophilia A can be used to test the efficacy of strategies for inhibitor suppression, but the differences in the immune systems of mice and humans limit its utility. To address this shortcoming, we established a humanized NOD/SCID-IL2rγnull hemophilia A (hu-NSG-HA) mouse model with a severely deficient mouse immune system presenting a patient's adapted immune cells. Through intrasplenic injection with patient inhibitor-positive peripheral blood mononuclear cells (PBMCs), utilizing an adeno-associated viral delivery system expressing human BLyS, and regular FVIII challenge, human C19+ B cells were expanded in vivo to secrete anti-FVIII antibodies. Both the inhibitor and the human anti-FVIII IgG, including the predominant subclasses (IgG1 and IgG4) present in the majority of inhibitor patients, were detected in the mouse model. We further segregated and expanded the different clones of human anti-FVIII–secreting cells through subsequent transplantation of splenocytes derived from hu-NSG-HA mice into another NSG-HA mouse. By transplanting a patient's PBMCs into the NSG-HA mouse model, we demonstrated the success of reintroducing a strong anti-FVIII immune response for a short period in mice with the immune systems of inhibitor-positive patients. Our results demonstrate a potential tool for directly obtaining functional human-derived antigen-specific antibodies and antibody-secreting cells, which may have therapeutic value for testing patient-specific immune responses to treatment options to assist in clinical decisions. • Treatment of hemophilia A with inhibitors remains a challenge with current regimens. • We created the first mouse model harboring patient-specific adaptive immunity. • Patient-specific B cells and anti-FVIII inhibitors can be induced in the model. • Humanized mice provide a testing platform to personalize therapeutic options. [ABSTRACT FROM AUTHOR]

NaV1.7, a voltage-gated sodium channel, induces chronic pain. Current research on the discovery of inhibitors against NaV1.7 is advancing with the hope of treating chronic pain conditions in patients suffering from various diseases. However, to characterize NaV1.7 and inhibitor interactions, a higher yield of expression is required to obtain sufficient protein. Molecular cloning of hNaV1.7 was done using the homologous recombination method in our novel S. cerevisiae-based D-crypt™ platform. The platform was designed for the high-yield production of 'difficult-to-express' proteins (DTE-Ps) up to 500 L. The changes in the cell membrane potential of hNaV1.7 were determined using a fluorescence resonance energy transfer (FRET) assay with tetracaine (hNaV1.7 inhibitor). Expression analysis of hNaV1.7 showed 2 bands of size 250 and 280 kDa that were absent in untransfected yeast cells. Immunofluorescence images revealed the presence of hNaV1.7 on the membrane of hNaV1.7-expressing yeast cells. Tetracaine exhibited concentration-dependent inhibition of the FRET ratio, with the order of potency (IC50 = 0.46 µM) being approximately the same as previously reported, suggesting the functionality of the channel protein expressed in the D-crypt™ platform. Cloning of NaV1.7 in the D-crypt™ platform will help to scale up the production of channel proteins, which will ultimately help in the structural and functional characterization of the binding interactions between toxins and the NaV1.7 channel to identify more specific NaV1.7 inhibitors. [ABSTRACT FROM AUTHOR]

Introduction: Adverse reactions are relatively common during peanut oral immunotherapy. To reduce the risk to the patient, some researchers have proposed modifying the allergen to reduce IgE reactivity, creating a putative hypoallergen. Analysis of recently cloned human IgG from patients treated with peanut immunotherapy suggested that there are three common conformational epitopes for the major peanut allergen Ara h 2. We sought to test if structural information on these epitopes could indicate mutagenesis targets for designing a hypoallergen and evaluated the reduction in IgE binding via immunochemistry and a mouse model of passive cutaneous anaphylaxis (PCA). Methods: X‐ray crystallography characterized the conformational epitopes in detail, followed by mutational analysis of key residues to modify monoclonal antibody (mAb) and serum IgE binding, assessed by ELISA and biolayer interferometry. A designed Ara h 2 hypoallergen was tested for reduced vascularization in mouse PCA experiments using pooled peanut allergic patient serum. Results: A ternary crystal structure of Ara h 2 in complex with patient antibodies 13T1 and 13T5 was determined. Site‐specific mutants were designed that reduced 13T1, 13T5, and 22S1 mAbs binding by orders of magnitude. By combining designed mutations from the three major conformational bins, a hexamutant (Ara h 2 E46R, E89R, E97R, E114R, Q146A, R147E) was created that reduced IgE binding in serum from allergic patients. Further, in the PCA model where mice were primed with peanut allergic patient serum, reactivity upon allergen challenge was significantly decreased using the hexamutant. Conclusion: These studies demonstrate that prior knowledge of common conformational epitopes can be used to engineer reduced IgE reactivity, an important first step in hypoallergen design. [ABSTRACT FROM AUTHOR]

Raëlism is a controversial new religious movement that originated in France in the 1970s and has since spread around the world. The movement's core tenets are rooted in science, technology, and freedom and can be summarized as a blend of science and spirituality, millenarianism, revelation, story, pseudoscience, and reinterpretation. Raëlians believe in the extraterrestrial origin of humanity and promote the pursuit of higher consciousness, inner peace, individual empowerment, and peace-building. To this end, the movement offers a series of seminars that aim to provide a transformative experience for participants and leads various social initiatives and projects at a global level. Although Raëlism has gained popularity among some individuals, it has also been the subject of substantial criticism and controversy, particularly because of its unconventional views on sexuality and its alleged support for human cloning and genetic engineering. Nonetheless, the movement continues to be a subject of interest and scrutiny for scholars, journalists, and the curious. This article provides an overview of the history, ideology, and practices of the Raëlian Movement and its relationship with transhumanism. [ABSTRACT FROM AUTHOR]

This paper addresses the issue of the loss of the real in the fiction of Kazuo Ishiguro as a contemporary author whose thought line is still in progress. His approach to this issue is anti-capitalist as he questions the so-called scientific advancement led by cash-oriented capitalists and industrialists. His writing seeks to strip the veil of the murderous nature of this kind of science. He blames it for killing the real and creating a world of simulations. He animadverts upon dystopian spaces where he holds postmodern scientific knowledge responsible for the digression of the natural course of life and lays bare the secrets behind the replacement of the real with the simulated by drawing attention to such debatable topics as human cloning, cosmetic surgeries and artificial intelligence. In terms of methodology of analysis, this paper is primarily based on a close examination of the author’s literary texts: two novels (Never Let Me Go and Klara and the Sun) and three short stories from Nocturnes (“Malvern Hills”, “Nocturne” and “Crooner”). Postmodernist concepts have been of great relevance to the analysis of these texts, for his fiction could not be approached in isolation from the realities of the postmodern era where it’s produced. Equally, bearing in mind the author’s socio-ethnic and historical background, the society where he lives and the politico-cultural transformations of the world aTher the Second World War plays an important role in the analysis of his texts.

Human milk fat globule epidermal growth factor-factor VIII (MFG-E8) functions as a bridging molecule to promote the removal of dying cells by professional phagocytes. E. coli-expressed histidine-tagged recombinant human MFG-E8 (rhMFG-E8) is protective in various disease conditions. However, due to improper recombinant protein glycosylation, misfolding and the possibility of antigenicity, E. coli-expressed histidine-tagged rhMFG-E8 is unsuitable for human therapy. Therefore, we hypothesize that human cell-expressed, tag-free rhMFG-E8 will have suitable structural and functional properties to be developed as a safe and effective novel biologic to treat inflammatory diseases including radiation injury. We produced a new tag-free rhMFG-E8 protein by cloning the human MFG-E8 full-length coding sequence without any fusion tag into a mammalian vector and expressed it in HEK293-derived cells. The construct includes the leader sequence of cystatin S to maximize secretion of rhMFG-E8 into the culture medium. After purification and confirmation of the protein identity, we first evaluated its biological activity in vitro. We then determined its efficacy in vivo utilizing an experimental rodent model of radiation injury, i.e., partial body irradiation (PBI). HEK293 cell supernatant containing tag-free rhMFG-E8 protein was concentrated, purified, and rhMFG-E8 was verified by SDS-PAGE with the standard human MFG-E8 loaded as control and, mass spectrometry followed by analysis using MASCOT for peptide mass fingerprint. The biological activity of human cell-expressed tag-free rhMFG-E8 was superior to that of E. coli-expressed His-tagged rhMFG-E8. Toxicity, stability, and pharmacokinetic studies indicate that tag-free rhMFG-E8 is safe, highly stable after lyophilization and long-term storage, and with a terminal elimination half-life in circulation of at least 1.45 h. In the 15 Gy PBI model, a dose-dependent improvement of the 30-day survival rate was observed after tag-free rhMFG-E8 treatment with a 30-day survival of 89%, which was significantly higher than the 25% survival in the vehicle group. The dose modification factor (DMF) of tag-free rhMFG-E8 calculated using probit analysis was 1.058. Tag-free rhMFG-E8 also attenuated gastrointestinal damage after PBI suggesting it as a potential therapeutic candidate for a medical countermeasure for radiation injury. Our new human cell-expressed tag-free rhMFG-E8 has proper structural and functional properties to be further developed as a safe and effective therapy to treat victims of severe acute radiation injury. [ABSTRACT FROM AUTHOR]

Genome editing in human pluripotent stem cells using programmable nucleases makes it possible to create models of hereditary pathologies using directed transgenesis, gene knockout, and replacement of individual nucleotides in DNA sequences. Using CRISPR/SpCas9-mediated homologous recombination at the AAVS1 locus, clones of human induced pluripotent stem cells (iPSCs) ICGi022-A (Malakhova et al., 2020) were obtained, which carry transgenes of two variants of the nuclease AsCas12a (also known as AsCpf1), recognizing different PAM consensuses, and the reverse doxycycline transgene-dependent transactivator M2rtTA. For each AsCas12a variant, the lines ICGi022-A-6 (AsCas12a, PAM 5'-TTTV-3') and ICGi022-A-7 (AsCas12a, PAM 5'-TYCV-3') were obtained. Using Western blot analysis, it was shown that the addition of doxycycline to the culture medium causes activation of the expression of AsCas12a(TTTV) and AsCas12a(TYCV) proteins. The resulting transgenic iPSC clones were subjected to molecular and cytogenetic analysis. Using quantitative PCR and immunocytochemical analysis, it was shown that they have a high level of mRNA expression of gene markers of pluripotent cells, namely OCT4, NANOG, and SOX2, as well as specific expression of protein markers OCT4, SOX2, SSEA-4, and TRA-1-60. In addition, using iPSCs spontaneous differentiation into embryoid bodies, it was found that transgenic clones can give derivatives of all three primitive germ layers: ectoderm, mesoderm, and endoderm. Cytogenetic analysis showed that transgenic iPSC clones have a normal karyotype, 46,XX. [ABSTRACT FROM AUTHOR]

CD8+ cytotoxic T lymphocytes (CTLs) play a crucial role in anti-tumor immunity. In a previous study, we identified a subset of murine effector CTLs expressing the hepatocyte growth factor (HGF) receptor, c-Met (c-Met+ CTLs), that are endowed with enhanced cytolytic capacity. HGF directly inhibited the cytolytic function of c-Met+ CTLs, both in 2D in vitro assays and in vivo, leading to reduced T cell responses against metastatic melanoma. To further investigate the role of c-Met+ CTLs in a three-dimensional (3D) setting, we studied their function within B16 melanoma spheroids and examined the impact of cell–cell contact on the modulation of inhibitory checkpoint molecules' expression, such as KLRG1, PD-1, and CTLA-4. Additionally, we evaluated the cytolytic capacity of human CTL clones expressing c-Met (c-Met+) and compared it to c-Met− CTL clones. Our results indicated that, similar to their murine counterparts, c-Met+ human CTL clones exhibited increased cytolytic activity compared to c-Met− CTL clones, and this enhanced function was negatively regulated by the presence of HGF. Taken together, our findings highlight the potential of targeting the HGF/c-Met pathway to modulate CTL-mediated anti-tumor immunity. This research holds promise for developing strategies to enhance the effectiveness of CTL-based immunotherapies against cancer. [ABSTRACT FROM AUTHOR]

N‑Acetyl‑d‑neuraminic acid (Neu5Ac) is the crucial compound for the chemical synthesis of antiflu medicine Zanamivir. Chemoenzymatic synthesis of Neu5Ac involves N-acetyl-d-glucosamine 2-epimerase (AGE)-catalyzed epimerization of N-acetyl-d-glucosamine (GlcNAc) to N-acetyl-d-mannosamine (ManNAc), and aldolase-catalyzed condensation between ManNAc and pyruvate. Host optimization plays an important role in the whole-cell biotransformation of value-added compounds. In this study, via single-plasmid biotransformation system, we showed that the AGE gene BT0453, cloned from human gut microorganism Bacteroides thetaiotaomicron VPI-5482, showed the highest biotransformation yield among the AGE genes tested; and there is no clear Neu5Ac yield difference between the BT0453 coupled with one aldolase coding nanA gene and two nanA genes. Next, Escherichia coli chromosomal genes involved in substrate degradation, product exportation and pH change were deleted via recombineering and CRISPR/Cas9. With the final E. coli BL21(DE3) ΔnanA Δnag ΔpoxB as host, a significant 16.5% yield improvement was obtained. Furthermore, precursor (pyruvate) feeding resulted in 3.2% yield improvement, reaching 66.8% molar biotransformation. The result highlights the importance of host optimization, and set the stage for further metabolic engineering of whole-cell biotransformation of Neu5Ac. [ABSTRACT FROM AUTHOR]

The migratory ability of microglia facilitates their rapid transport to a site of injury to kill and remove pathogens. However, the effect of Treponema pallidum membrane proteins on microglia migration remains unclear. The effect of Tp47 on the migration ability and autophagy and related mechanisms were investigated using the human microglial clone 3 cell line. Tp47 inhibited microglia migration, the expression of autophagy‐associated protein P62 decreased, the expression of Beclin‐1 and LC3‐II/LC3‐I increased, and the autophagic flux increased in this process. Furthermore, autophagy was significantly inhibited, and microglial cell migration was significantly increased after neutralisation with an anti‐Tp47 antibody. In addition, Tp47 significantly inhibited the expression of p‐PI3K, p‐AKT, and p‐mTOR proteins, and the sequential activation of steps in the PI3K/AKT/mTOR pathways effectively prevented Tp47‐induced autophagy. Moreover, Tp47 significantly inhibited the expression of p‐FOXO1 protein and promoted FOXO1 nuclear translocation. Inhibition of FOXO1 effectively suppressed Tp47‐induced activation of autophagy and inhibition of migration. Treponema pallidum membrane protein Tp47‐induced autophagy and inhibited cell migration in HMC3 Cells via the PI3K/AKT/FOXO1 pathway. These data will contribute to understanding the mechanism by which T. pallidum escapes immune killing and clearance after invasion into the central nervous system. [ABSTRACT FROM AUTHOR]

Reservoir host associations have been observed among and within Borrelia genospecies, and host complement-mediated killing is a major determinant in these interactions. In North America, only a subset of Borrelia burgdorferi lineages cause the majority of disseminated infections in humans. We hypothesize that differential resistance to human complement-mediated killing may be a major phenotypic determinant of whether a lineage can establish systemic infection. As a corollary, we hypothesize that borreliacidal action may differ among human subjects. To test these hypotheses, we isolated primary B. burgdorferi clones from field-collected ticks and determined whether the killing effects of human serum differed among those clones in vitro and/or whether these effects were consistent among human sera. Clones associated with human invasiveness did not show higher survival in human serum compared to noninvasive clones. These results indicate that differential complement-mediated killing of B. burgdorferi lineages is not a determinant of invasiveness in humans. Only one significant difference in the survivorship of individual clones incubated in different human sera was detected, suggesting that complement-mediated killing of B. burgdorferi is usually similar among humans. Mechanisms other than differential human complement-mediated killing of B. burgdorferi lineages likely explain why only certain lineages cause the majority of disseminated human infections. [ABSTRACT FROM AUTHOR]

T-cell receptor (TR) diversity of the variable domains is generated by recombination of both the alpha (TRA) and beta (TRB) chains. The textbook process of TRB chain production starts with TRBD and TRBJ gene rearrangement, followed by the rearrangement of a TRBV gene to the partially rearranged D-J gene. Unsuccessful V-D-J TRB rearrangements lead to apoptosis of the cell. Here, we performed deep sequencing of the poorly explored pool of partial TRBD1-TRBD2 rearrangements in T-cell genomic DNA. We reconstructed full repertoires of human partial TRBD1-TRBD2 rearrangements using novel sequencing and validated them by detecting V-D-J recombination-specific byproducts: excision circles containing the recombination signal (RS) joint 5'D2-RS - 3'D1-RS. Identified rearrangements were in compliance with the classical 12/23 rule, common for humans, rats, and mice and contained typical V-D-J recombination footprints. Interestingly, we detected a bimodal distribution of D-D junctions indicating two active recombination sites producing long and short D-D rearrangements. Long TRB D-D rearrangements with two D-regions are coding joints D1-D2 remaining classically on the chromosome. The short TRB D-D rearrangements with no D-region are signal joints, the coding joint D1-D2 being excised from the chromosome. They both contribute to the TRB V-(D)-J combinatorial diversity. Indeed, short D-D rearrangements may be followed by direct V-J2 recombination. Long D-D rearrangements may recombine further with J2 and V genes forming partial D1-D2-J2 and then complete V-D1-D2-J2 rearrangement. Productive TRB V-D1-D2-J2 chains are present and expressed in thousands of clones of human antigen-experienced memory T cells proving their capacity for antigen recognition and actual participation in the immune response. [ABSTRACT FROM AUTHOR]

In response to the Journal of Religious Ethics (JRE) editors' request for reflections on "how religious ethicists have interacted with, and ought to interact with, public policy decision makers," this essay focuses on doing religious ethics in the context of doing public bioethics, especially through participating in public bioethics bodies (PBBs) established to provide advice to public policymakers in what might be called "mediated advocacy." Drawing heavily on the author's experience as a member of and a consultant to several PBBs, it features case studies of PBBs deliberating about and recommending public policies to address the scarcity of postmortem organs for transplantation, the equitable allocation of COVID‐19 vaccine, cloning humans, and human embryonic stem cell research. [ABSTRACT FROM AUTHOR]

The online space has continued to be a platform for not only private and mundane discussions, but also a tribune for voicing critical political and national opinions. Nigerians and the international community have employed online media, as well as other media platforms to articulate their thoughts on the claims which favoured the possibility of the demise of the immediate past president of Nigeria, President Muhammadu Buhari, after a prolonged illness in the year 2007 and the ploy of a Jubril of Sudan clone in his place. This study examined the pragmatic strategies employed in the online articles on the national dilemma caused by the Buhari/Jubril claims and refutals, in response to the lacuna in the literature on such analytical investigations on the subject. An online article titled, "Buhari: The real, the fake and the dead" authored by a Nigerian writer, Tunde Odesola retrieved from the online page of The Punch Newspaper on December 3, 2018, served as data for the study. The article was analysed using insights from the Stance Theory and the Pragmatics in general. The analysis revealed eight pragmatic strategies utilised by the writer, namely Biblical allusion, evaluative stance and positioning, epistemic stance and evocation of sarcasm, metaphors, and derogative labelling as anti-Jubrilist positioning, epistemic stance, and evocation of (dis)alignment, antithetical evocations, evaluative stance and berating, evaluative stance and justifying. These result in a pragmatic reconstruction of the readers' views on the issue. [ABSTRACT FROM AUTHOR]

This scholarly article delves into the multifaceted domains of human cloning, encompassing its biological underpinnings, ethical dimensions, and broader societal implications. The exposition commences with a succinct historical and contextual overview of human cloning, segueing into an in-depth exploration of its biological intricacies. Central to this biological scrutiny is a comprehensive analysis of somatic cell nuclear transfer (SCNT) and its assorted iterations. The accomplishments and discoveries in cloning technology, such as successful animal cloning operations and advances in the efficiency and viability of cloned embryos, are reviewed. Future improvements, such as reprogramming procedures and gene editing technology, are also discussed. The discourse extends to ethical quandaries intrinsic to human cloning, entailing an extensive contemplation of values such as human dignity, autonomy, and safety. Furthermore, the ramifications of human cloning on a societal plane are subjected to scrutiny, with a dedicated emphasis on ramifications encompassing personal identity, kinship connections, and the fundamental notion of maternity. Culminating the analysis is a reiteration of the imperative to develop and govern human cloning technology judiciously and conscientiously. Finally, it discusses several ethical and practical issues, such as safety concerns, the possibility of exploitation, and the erosion of human dignity, and emphasizes the significance of carefully considering these issues. [ABSTRACT FROM AUTHOR]

Ethical and theological debates concerning human cloning are getting more intense as cloning technology continues to develop. This has led to debates originating from both secular ethics and Christian theological perspectives. This paper aims to coordinate the debate according to these two perspectives. It will draw from the positions of Christian theology, the position of the United Nations Universal Declaration of Human Rights, and the positions of scientific authorities. It will conclude by stressing the importance of regulating new technologies in a manner which recognizes the shared ethico-theological concerns for human dignity, rights, freedoms, and the moral growth of humanity. [ABSTRACT FROM AUTHOR]

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This research article, using the lens of Bioconservatism, critically analyses the ethical concerns raised by Sangu Mandanna's novel The Lost Girl involving human cloning. This study delves into the complex ethical concerns surrounding cloning technology and considers its effects on identity, parent-child relationship, societal acceptance, and right to existence through a detailed analysis of the story of the novel, its characters, and themes. The aim of this article is to elicit critical debates on the moral dilemmas that emerge when science and ethics collide. [ABSTRACT FROM AUTHOR]

Expression cloning of fully human monoclonal antibodies (hmAbs) is seeing powerful utility in the field of vaccinology, especially for elucidating vaccine-induced B-cell responses and novel vaccine candidate antigen discovery. Precision of the hmAb cloning process relies on efficient isolation of hmAb-producing plasmablasts of interest. Previously, a novel immunoglobulin-capture assay (ICA) was developed, using single protein vaccine antigens, to enhance the pathogen-specific hmAb cloning output. Here, we report a novel modification of this single-antigen ICA using formalin-treated, fluorescently stained whole cell suspensions of the human bacterial invasive pathogens, Streptococcus pneumoniae and Neisseria meningitidis. Sequestration of IgG secreted by individual vaccine antigen-specific plasmablasts was achieved by the formation of an anti-CD45-streptavidin and biotin anti-IgG scaffold. Suspensions containing heterologous pneumococcal and meningococcal strains were then used to enrich for polysaccharide- and protein antigen-specific plasmablasts, respectively, during single cell sorting. Following application of the modified whole-cell ICA (mICA), ~61% (19/31) of anti-pneumococcal polysaccharide hmAbs were cloned compared to 14% (8/59) obtained using standard (non-mICA) methods - representing a ~4.4-fold increase in hmAb cloning precision. A more modest ~1.7-fold difference was obtained for anti-meningococcal vaccine hmAb cloning; ~88% of hmAbs cloned via mICA versus ~53% cloned via the standard method were specific for a meningococcal surface protein. VDJ sequencing revealed that cloned hmAbs reflected an anamnestic response to both pneumococcal and meningococcal vaccines; diversification within hmAb clones occurred by positive selection for replacement mutations. Thus, we have shown successful utilization of whole bacterial cells in the ICA protocol enabling isolation of hmAbs targeting multiple disparate epitopes, thereby increasing the power of approaches such as reverse vaccinology 2.0 (RV 2.0) for bacterial vaccine antigen discovery. [ABSTRACT FROM AUTHOR]

Answering climate change challenges, the paper proposes an intelligent decision support system (DSS) for the management of green-blue infrastructure (GBI). Addressing the gaps identified in other studies, the designed DSS incorporates four key elements: 1/interdisciplinary collaboration among all stakeholders 2/inclusion of practical operation and maintenance activities, 3/main components of distributed DSS, with practical examples of use, 4/consideration of conditions specific to the location. The multi-layered DSS architecture can be implemented as a unified platform that provides a comprehensive, customizable, and flexible framework based on AI tools, big data and analytics, edge computing, cloud, and mobile, IIoT, and biometric system tools. The use of cobots and digital clones alongside humans results in the implementation of hybrid human-machine units. DSS for GBI increases decision-making capacity and can serve as a foundation for the implementation of similar systems by governments and local communities to build sustainable and resilient communities. [ABSTRACT FROM AUTHOR]

But arguably the most important legacy was the canonization of a version of cloning's history shorn of all the contingencies which Crowe has now so meticulously and thoughtfully restored. No biological technique has had a better run in speculative fiction than cloning. [Extracted from the article]

We propose a novel approach for robotic industrial insertion tasks using the Programming by Demonstration technique. Our method allows robots to learn a high-precision task by observing human demonstration once, without requiring any prior knowledge of the object. We introduce an Imitated-to-Finetuned approach that generates imitated approach trajectories by cloning the human hand's movements and then fine-tunes the goal position with a visual servoing approach. To identify features on the object used in visual servoing, we model object tracking as the moving object detection problem, separating each demonstration video frame into the moving foreground that includes the object and demonstrator's hand and the static background. Then a hand keypoints estimation function is used to remove the redundant features on the hand. The experiment shows that the proposed method can make robots learn precision industrial insertion tasks from a single human demonstration. [ABSTRACT FROM AUTHOR]

Methicillin-resistant Staphylococcus aureus (MRSA) is a strain of pathogenic bacteria that is a major problem in the world's health. Due to their frequent interaction with humans, pets are one of the main risk factors for the spread of MRSA. The possibility for zoonotic transmission exists since frequently kept dogs and cats are prone to contract MRSA and act as reservoirs for spreading MRSA. The mouth, nose, and perineum are the primary locations of MRSA colonization, according to the findings of MRSA identification tests conducted on pets. The types of MRSA clones identified in cats and dogs correlated with MRSA clones infecting humans living in the same geographic area. A significant risk factor for the colonization or transmission of MRSA is human-pet contact. An essential step in preventing the spread of MRSA from humans to animals and from animals to humans is to keep hands, clothing, and floor surfaces clean. [ABSTRACT FROM AUTHOR]

TRPA1 (transient receptor potential ankyrin 1) is a nonselective Ca2+-permeable cation channel, which was originally cloned from human lung fibroblasts (HLFs). TRPA1-mediated Ca2+ entry is evoked by exposure to several chemicals, including allyl isothiocyanate (AITC), and a protective effect of TRPA1 activation in the development of cardiac fibrosis has been proposed. Yet the function of TRPA1 in TGF-β1 (transforming growth factor-β1)-driven fibroblast-to-myofibroblast differentiation and the development of pulmonary fibrosis remains elusive. TRPA1 expression and function were analyzed in cultured primary HLFs, and mRNA concentrations were significantly reduced after adding TGF-β1. Expression of genes encoding fibrosis markers (e.g., ACTA2, SERPINE1 [plasminogen activator inhibitor 1], FN1 [fibronectin], COL1A1 [type I collagen]) was increased after siRNA-mediated downregulation of TRPA1 mRNA in HLFs. Moreover, AITC-induced Ca2+ entry in HLFs was decreased after TGF-β1 treatment and by application of TRPA1 siRNAs, while AITC treatment alone did not reduce cell viability or enhance apoptosis. Most interestingly, AITC-induced TRPA1 activation augmented ERK1/2 (extracellular signal-regulated kinase 1/2) and SMAD2 linker phosphorylation, which might inhibit TGF-β-receptor signaling. Our results suggest an inhibitory function of TRPA1 channels in TGF-β1-driven fibroblast-to-myofibroblast differentiation. Therefore, activation of TRPA1 channels might be protective during the development of pulmonary fibrosis in patients. [ABSTRACT FROM AUTHOR]

The Department of Defense recently began an effort to improve and standardize virus challenge materials and efficacy determination strategies for testing therapeutics and vaccines. This includes stabilization of virus genome sequences in cDNA form where appropriate, use of human-derived virus isolates, and noninvasive strategies for determination of challenge virus replication. Eventually, it is desired that these approaches will satisfy the FDA "Animal Rule" for licensure, which substitutes animal efficacy data when human data are unlikely to be available. To this end, we created and examined the virulence phenotype of cDNA clones of prototypic human infection-derived strains of the alphaviruses, Venezuelan (VEEV INH9813), eastern (EEEV V105) and western (WEEV Fleming) equine encephalitis viruses, and created fluorescent and luminescent reporter expression vectors for evaluation of replication characteristics in vitro and in vivo. Sequences of minimally passaged isolates of each virus were used to synthesize full-length cDNA clones along with a T7 transcription promoter-based bacterial propagation vector. Viruses generated from the cDNA clones were compared with other "wild type" strains derived from cDNA clones and GenBank sequences to identify and eliminate putative tissue culture artifacts accumulated in the cell passaged biological stocks. This was followed by examination of aerosol and subcutaneous infection and disease in mouse models. A mutation that increased heparan sulfate binding was identified in the VEEV INH9813 biological isolate sequence and eliminated from the cDNA clone. Viruses derived from the new human isolate cDNA clones showed similar mouse virulence to existing clone-derived viruses after aerosol or subcutaneous inoculation. [ABSTRACT FROM AUTHOR]

Autoantibodies targeting brain antigens can mediate a wide range of neurological symptoms ranging from epileptic seizures to psychosis to dementia. Although earlier experimental work indicated that autoantibodies can be directly pathogenic, detailed studies on disease mechanisms, biophysical autoantibody properties, and target interactions were hampered by the availability of human material and the paucity of monospecific disease-related autoantibodies. The emerging generation of patient-derived monoclonal autoantibodies (mAbs) provides a novel platform for the detailed characterization of immunobiology and autoantibody pathogenicity in vitro and in animal models. This Feature Review focuses on recent advances in mAb generation and discusses their potential as powerful scientific tools for high-resolution imaging, antigenic target identification, atomic-level structural analyses, and the development of antibody-selective immunotherapies. Advances in the cloning of human monoclonal autoantibodies (mAbs) and their recombinant production in theoretically unlimited amounts have sparked new opportunities to study pathogenic mechanisms in vitro and in vivo , thereby providing direct proof of autoantibody pathogenicity in autoimmune encephalopathies. Human mAbs – in contrast to polyclonal sera or cerebrospinal fluid samples – are valuable scientific tools for novel experimental approaches ranging from super-resolution microscopy to structural determination of antibody–antigen interaction at the atomic level. Only analyses at the monoclonal level have demonstrated that, despite binding the same molecules, mAbs group into subclasses with diverse epitopes, isotypes, and distinct functional effects. Human mAbs can guide the development of highly selective antibody-specific immunotherapies for autoimmune encephalopathies. [ABSTRACT FROM AUTHOR]