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1. Proactive vaccination using multiviral Quartet Nanocages to elicit broad anti-coronavirus responses

Author

Hills, Rory A., Tan, Tiong Kit, Cohen, Alexander A., Keeffe, Jennifer R., Keeble, Anthony H., Gnanapragasam, Priyanthi N. P., Storm, Kaya N., Rorick, Annie V., West, Jr., Anthony P., Hill, Michelle L., Liu, Sai, Gilbert-Jaramillo, Javier, Afzal, Madeeha, Napier, Amy, Admans, Gabrielle, James, William S., Bjorkman, Pamela J., Townsend, Alain R., and Howarth, Mark R.

Published
2024
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3. Metabolic remodelling defines cellular subtypes of human neuronal progenitors and their susceptibility to Zika virus replication

Author

Gilbert Jaramillo, Javier Andrés, James, William, Molnar, Zoltan, and Clarke, Kieran

Subjects
Virology, Neurodevelopment, Energy metabolism
Abstract

Maternal infections by Zika virus (ZIKV) during the first trimester of pregnancy result in either abortion or severe damage to the developing brain in 8-12% of cases. However, the route of ZIKV to the developing brain remains unclear. Emerging work suggests ZIKV replication imposes energetic demands on the developing brain that overwrite the highly orchestrated energetic processes required for normal cellular growth, proliferation, and differentiation. Energetic dysfunction could be attributed as the main cause for ZIKV-induced detrimental brain damage in new-borns yet, there is no clear evidence on the altered energetic pathway(s) and/or substrate(s) required to sustain ZIKV replication. The work presented in this thesis showed the characterization of 2D in vitro cultures of cortical neuronal progenitor cells (hi-NPCs) and tissue-resident-like macrophages (hiPSC-MPh) derived from human pluripotent stem cells as models of the early fetal brain to better understand the pathogenesis of ZIKV. Analysis of the preferred energetic pathway in hi-NPCs distinguished two distinct subtypes of brain progenitors resembling populations with characteristics of quiescent radial-glia-like cells or proliferative intermediate neuronal progenitors. ZIKV infection in hi-NPCs revealed a subtype-specific permissibility to infection with metabolic implications resulting in reprogramming towards an increased cytosolic glucose metabolism. This was not observed in hiPSC-MPh as these cells showed less permissibility to ZIKV infection than hi-NPCs yet, the low rates of viral production in hiPSC-MPh were sufficient to disseminate virions to hi-NPCs, highlighting a potential route for maternal-fetal transmission of ZIKV. Further analysis using immune-metabolic compromised RSAD2-KO hiPSC-MPh showed that cytosolic glycolysis may underlie mechanisms for high rates of ZIKV replication. However, more research and validation of the current results are required to clearly elucidate whether energy intermediates can diminish ZIKV replication for potential therapeutics. These were not explored in detail due to the disruptions caused by the COVID-19 pandemic, instead this thesis includes an isolated chapter on SARS-CoV-2 because of the change in research projects. The small research on SARS-CoV-2 hypothesises that macrophages do not negatively contribute to severe COVID-19 by the triggering the cytokine storm but have a dual role by decreasing SARS-CoV-2 replication kinetics by the degradation of circulating virions. Herein, I showed that macrophages, both peripheral (MDM) and hiPSC-MPh, were not productively infected by SARS-CoV-2 virions after direct (cell-free) or indirect exposure (cell-to-cell transfer). Instead, these cells phagocytized virions for phagolysosome degradation and decreased the viral replication kinetics by reducing the amount of output virions that can infect susceptible cells. This suggested a potential protective role of macrophages against the dissemination of SARS-CoV-2 within infected tissues.

Published
2022

9. Pepper catalase: a broad analysis of its modulation during fruit ripening and by nitric oxide.

Author

González-Gordo, Salvador, López-Jaramillo, Javier, Rodríguez-Ruiz, Marta, Taboada, Jorge, Palma, José M., and Corpas, Francisco J.

Subjects
*CAPSICUM annuum, *FRUIT ripening, *CATALASE, *PEPPERS, *NITRIC oxide, *POLYACRYLAMIDE gel electrophoresis, *PLANT cell microbodies
Abstract

Catalase is a major antioxidant enzyme located in plant peroxisomes that catalyzes the decomposition of H2O2. Based on our previous transcriptomic (RNA-Seq) and proteomic (iTRAQ) data at different stages of pepper (Capsicum annuum L.) fruit ripening and after exposure to nitric oxide (NO) enriched atmosphere, a broad analysis has allowed us to characterize the functioning of this enzyme. Three genes were identified, and their expression was differentially modulated during ripening and by NO gas treatment. A dissimilar behavior was observed in the protein expression of the encoded protein catalases (CaCat1–CaCat3). Total catalase activity was down-regulated by 50% in ripe (red) fruits concerning immature green fruits. This was corroborated by non-denaturing polyacrylamide gel electrophoresis, where only a single catalase isozyme was identified. In vitro analyses of the recombinant CaCat3 protein exposed to peroxynitrite (ONOO−) confirmed, by immunoblot assay, that catalase underwent a nitration process. Mass spectrometric analysis identified that Tyr348 and Tyr360 were nitrated by ONOO−, occurring near the active center of catalase. The data indicate the complex regulation at gene and protein levels of catalase during the ripening of pepper fruits, with activity significantly down-regulated in ripe fruits. Nitration seems to play a key role in this down-regulation, favoring an increase in H2O2 content during ripening. This pattern can be reversed by the exogenous NO application. While plant catalases are generally reported to be tetrameric, the analysis of the protein structure supports that pepper catalase has a favored quaternary homodimer nature. Taken together, data show that pepper catalase is down-regulated during fruit ripening, becoming a target of tyrosine nitration, which provokes its inhibition. [ABSTRACT FROM AUTHOR]

Published
2024
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12. Structural basis for the neutralization of SARS-CoV-2 by an antibody from a convalescent patient

Author

Zhou, Daming, Duyvesteyn, Helen M. E., Chen, Cheng-Pin, Huang, Chung-Guei, Chen, Ting-Hua, Shih, Shin-Ru, Lin, Yi-Chun, Cheng, Chien-Yu, Cheng, Shu-Hsing, Huang, Yhu-Chering, Lin, Tzou-Yien, Ma, Che, Huo, Jiandong, Carrique, Loic, Malinauskas, Tomas, Ruza, Reinis R., Shah, Pranav N. M., Tan, Tiong Kit, Rijal, Pramila, Donat, Robert F., Godwin, Kerry, Buttigieg, Karen R., Tree, Julia A., Radecke, Julika, Paterson, Neil G., Supasa, Piyada, Mongkolsapaya, Juthathip, Screaton, Gavin R., Carroll, Miles W., Gilbert-Jaramillo, Javier, Knight, Michael L., James, William, Owens, Raymond J., Naismith, James H., Townsend, Alain R., Fry, Elizabeth E., Zhao, Yuguang, Ren, Jingshan, Stuart, David I., and Huang, Kuan-Ying A.

Published
2020
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13. Neutralizing nanobodies bind SARS-CoV-2 spike RBD and block interaction with ACE2

Author

Huo, Jiandong, Le Bas, Audrey, Ruza, Reinis R., Duyvesteyn, Helen M. E., Mikolajek, Halina, Malinauskas, Tomas, Tan, Tiong Kit, Rijal, Pramila, Dumoux, Maud, Ward, Philip N., Ren, Jingshan, Zhou, Daming, Harrison, Peter J., Weckener, Miriam, Clare, Daniel K., Vogirala, Vinod K., Radecke, Julika, Moynié, Lucile, Zhao, Yuguang, Gilbert-Jaramillo, Javier, Knight, Michael L., Tree, Julia A., Buttigieg, Karen R., Coombes, Naomi, Elmore, Michael J., Carroll, Miles W., Carrique, Loic, Shah, Pranav N. M., James, William, Townsend, Alain R., Stuart, David I., Owens, Raymond J., and Naismith, James H.

Published
2020
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15. Psicología del trabajo y de las organizaciones. Reflexiones y experiencias de investigación

Author

Aguilar Bustamante, María Constanza, primary, Rentería Pérez, Erico, additional, Peiró, José María, additional, Peralta Gómez, María Claudia, additional, Pulido Martínez, Hernán Camilo, additional, Vesga Rodríguez, Juan Javier, additional, Rosero Sarasty, Óscar Martín, additional, Fang Alandette, Yuly Patricia, additional, Valencia Patiño, Ricardo Antonio, additional, Mantilla, Fanny Teresa, additional, Castañeda Zapata, Delio Ignacio, additional, Enríquez Martínez, Álvaro, additional, Delgado Abella, Leonor Emilia, additional, García Álvarez, Claudia María, additional, Acosta Riaño, Carlos Alberto, additional, García Rubiano, Mónica, additional, Vélez Botero, Helena, additional, González Gutiérrez, Luis Felipe, additional, Giraldo Jaramillo, Javier Vicente, additional, Gómez Barrios, Ingrid Carolina, additional, Sierra Castellanos, Yolanda, additional, Rodríguez, Jeimy Andrea, additional, Guecha, Yenny Patricia, additional, Hermosa Rodríguez, Ángelica María, additional, Sandoval E., Marithza, additional, Caycedo E., Claudia, additional, Botero P., María M., additional, López L., Wilson, additional, Javela Delgado, Lady Grey, additional, Botero Posada, María Mercedes, additional, Cruz V., Julio Eduardo, additional, Quevedo Bermúdez, Martín, additional, Carvajal Marín, Luz Mery, additional, Botero Manzur, Eduardo, additional, Mejía López, Ana María, additional, Huertas Hernández, Olga Lucía, additional, Burbano Valente, Johanna, additional, Gómez, Irma Yaneth, additional, and González Herrera, Alvaro Giovanni, additional

Published
2020
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19. Real-Life Effectiveness and Tolerability of Brivaracetam in Focal to Bilateral and Primary Generalized Tonic-Clonic Seizures.

Author

Fonseca, Elena, Gifreu, Ariadna, Abril Jaramillo, Javier, Redondo Vergé, Luis, Quintana, Manuel, Lallana, Sofía, López-Maza, Samuel, Abraira, Laura, Campos-Fernández, Daniel, Santamarina, Estevo, Rodríguez Uranga, Juan, and Toledo, Manuel

Subjects
SEIZURES (Medicine), EPILEPSY, PEOPLE with epilepsy, PATIENT safety, DRUG resistance, DROWSINESS
Abstract

Purpose. Brivaracetam (BRV), an antiseizure medication indicated for focal-onset seizures, has shown efficacy in the treatment of focal to bilateral tonic-clonic seizures (FBTCS). We aimed to determine the effectiveness and safety of BRV in patients with FBTCS and generalized tonic-clonic seizures (GTCS). Methods. We performed a multicenter, retrospective, longitudinal study in adult patients with epilepsy who experienced at least one FBTCS or GTCS before starting BRV (baseline visit). Data were collected from consecutive outpatient visits over a 4-year period. All patients had been followed for at least 3 months before the baseline visit and completed a minimum follow-up of 3 months after starting BRV. Response (≥50% reduction in FBTCS/GTCS frequency) and retention rates, as well as seizure freedom and presence of adverse events at 3, 6, and 12 months, were recorded as outcome measures. Results. 114 patients were included (mean age 36.3 ± 18.0 years, 52% male, 36.6% genetic generalized epilepsy); 94 had a 12-month follow-up period. At 12 months' follow-up, the response rate was 83%, and 73.4% of patients were FBTCS/GTCS-free. Retention was 79% at 12 months. Adverse events occurred in 29.8% of patients, the most common being drowsiness (14.9%). No significant differences were found in response rates between FBTCS and GTCS. Drug resistance was independently associated with lower response and seizure freedom rates at follow-up. The absence of a titration period predicted seizure freedom and response at 3 months. Conclusions. BRV is an effective and well-tolerated treatment in patients with focal to bilateral tonic-clonic seizures and generalized tonic-clonic seizures. [ABSTRACT FROM AUTHOR]

Published
2023
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21. G‐quadruplex‐binding small molecules ameliorate C9orf72 FTD/ALS pathology in vitro and in vivo

Author

Simone, Roberto, Balendra, Rubika, Moens, Thomas G, Preza, Elisavet, Wilson, Katherine M, Heslegrave, Amanda, Woodling, Nathan S, Niccoli, Teresa, Gilbert‐Jaramillo, Javier, Abdelkarim, Samir, Clayton, Emma L, Clarke, Mica, Konrad, Marie‐Therese, Nicoll, Andrew J, Mitchell, Jamie S, Calvo, Andrea, Chio, Adriano, Houlden, Henry, Polke, James M, Ismail, Mohamed A, Stephens, Chad E, Vo, Tam, Farahat, Abdelbasset A, Wilson, W David, Boykin, David W, Zetterberg, Henrik, Partridge, Linda, Wray, Selina, Parkinson, Gary, Neidle, Stephen, Patani, Rickie, Fratta, Pietro, and Isaacs, Adrian M

Published
2018
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25. Broadly neutralizing aptamers to SARS-CoV-2: A diverse panel of modified DNA antiviral agents

Author

Gelinas, Amy D., primary, Tan, Tiong Kit, additional, Liu, Sai, additional, Jaramillo, Javier G., additional, Chadwick, James, additional, Harding, Adam C., additional, Zhang, Chi, additional, Ream, Brian E., additional, Chase, Chelsea N., additional, Otis, Matthew R., additional, Lee, Thomas, additional, Schneider, Daniel J., additional, James, William S., additional, and Janjic, Nebojsa, additional

Published
2023
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26. Multiviral Quartet Nanocages Elicit Broad Anti-Coronavirus Responses for Proactive Vaccinology

Author

Hills, Rory A., Tan, Tiong Kit, Cohen, Alexander A., Keeffe, Jennifer R., Keeble, Anthony H., Gnanapragasam, Priyanthi N. P., Storm, Kaya N., Hill, Michelle L., Liu, Sai, Gilbert-Jaramillo, Javier, Afzal, Madeeha, Napier, Amy, James, William S., Bjorkman, Pamela J., Townsend, Alain R., Howarth, Mark, Hills, Rory A., Tan, Tiong Kit, Cohen, Alexander A., Keeffe, Jennifer R., Keeble, Anthony H., Gnanapragasam, Priyanthi N. P., Storm, Kaya N., Hill, Michelle L., Liu, Sai, Gilbert-Jaramillo, Javier, Afzal, Madeeha, Napier, Amy, James, William S., Bjorkman, Pamela J., Townsend, Alain R., and Howarth, Mark

Abstract

Defending against future pandemics may require vaccine platforms that protect across a range of related pathogens. The presentation of multiple receptor-binding domains (RBDs) from evolutionarily-related viruses on a nanoparticle scaffold elicits a strong antibody response to conserved regions. Here we produce quartets of tandemly-linked RBDs from SARS-like betacoronaviruses coupled to the mi3 nanocage through a SpyTag/SpyCatcher spontaneous reaction. These Quartet Nanocages induce a high level of neutralizing antibodies against several different coronaviruses, including against viruses not represented on the vaccine. In animals primed with SARS-CoV-2 Spike, boost immunizations with Quartet Nanocages increased the strength and breadth of an otherwise narrow immune response. Quartet Nanocages are a strategy with potential to confer heterotypic protection against emergent zoonotic coronavirus pathogens and facilitate proactive pandemic protection.

Published
2023

27. Soybean (Glycine max L.) Lipoxygenase 1 (LOX 1) is modulated by nitric oxide and hydrogen sulfide: an in vitro approach

Author

European Commission, Ministerio de Ciencia, Innovación y Universidades (España), Agencia Estatal de Investigación (España), Junta de Andalucía, González-Gordo, Salvador, López-Jaramillo, Javier, Palma Martínez, José Manuel, Corpas, Francisco J., European Commission, Ministerio de Ciencia, Innovación y Universidades (España), Agencia Estatal de Investigación (España), Junta de Andalucía, González-Gordo, Salvador, López-Jaramillo, Javier, Palma Martínez, José Manuel, and Corpas, Francisco J.

Abstract

Hydrogen sulfide (H2S) and nitric oxide (NO) are two relevant signal molecules that can affect protein function throughout post-translational modifications (PTMs) such as persulfidation, S-nitrosation, metal-nitrosylation, and nitration. Lipoxygenases (LOXs) are a group of non-heme iron enzymes involved in a wide range of plant physiological functions including seed germination, plant growth and development, and fruit ripening and senescence. Likewise, LOXs are also involved in the mechanisms of response to diverse environmental stresses. Using purified soybean (Glycine max L.) lipoxygenase type 1 (LOX 1) and nitrosocysteine (CysNO) and sodium hydrosulfide (NaHS) as NO and H2S donors, respectively, the present study reveals that both compounds negatively affect LOX activity, suggesting that S-nitrosation and persulfidation are involved. Mass spectrometric analysis of nitrated soybean LOX 1 using a peroxynitrite (ONOO−) donor enabled us to identify that, among the thirty-five tyrosine residues present in this enzyme, only Y214 was exclusively nitrated by ONOO−. The nitration of Y214 seems to affect its interaction with W500, a residue involved in the substrate binding site. The analysis of the structure 3PZW demonstrates the existence of several tunnels that directly communicate the surface of the protein with different internal cysteines, thus making feasible their potential persulfidation, especially C429 and C127. On the other hand, the CysNO molecule, which is hydrophilic and bulkier than H2S, can somehow be accommodated throughout the tunnel until it reaches C127, thus facilitating its nitrosation. Overall, a large number of potential persulfidation targets and the ease by which H2S can reach them through the diffuse tunneling network could be behind their efficient inhibition.

Published
2023

28. Author Correction: Neutralizing nanobodies bind SARS-CoV-2 spike RBD and block interaction with ACE2

Author

Huo, Jiandong, Bas, Audrey Le, Ruza, Reinis R., Duyvesteyn, Helen M. E., Mikolajek, Halina, Malinauskas, Tomas, Tan, Tiong Kit, Rijal, Pramila, Dumoux, Maud, Ward, Philip N., Ren, Jingshan, Zhou, Daming, Harrison, Peter J., Weckener, Miriam, Clare, Daniel K., Vogirala, Vinod K., Radecke, Julika, Moynié, Lucile, Zhao, Yuguang, Gilbert-Jaramillo, Javier, Knight, Michael L., Tree, Julia A., Buttigieg, Karen R., Coombes, Naomi, Elmore, Michael J., Carroll, Miles W., Carrique, Loic, Shah, Pranav N. M., James, William, Townsend, Alain R., Stuart, David I., Owens, Raymond J., and Naismith, James H.

Published
2021
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29. Multiviral Quartet Nanocages Elicit Broad Anti-Coronavirus Responses for Proactive Vaccinology

Author

Hills, Rory A., primary, Tan, Tiong Kit, additional, Cohen, Alexander A., additional, Keeffe, Jennifer R., additional, Keeble, Anthony H., additional, Gnanapragasam, Priyanthi N.P., additional, Storm, Kaya N., additional, Hill, Michelle L., additional, Liu, Sai, additional, Gilbert-Jaramillo, Javier, additional, Afzal, Madeeha, additional, Napier, Amy, additional, James, William S., additional, Bjorkman, Pamela J., additional, Townsend, Alain R., additional, and Howarth, Mark, additional

Published
2023
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35. Author Correction: Neutralizing nanobodies bind SARS-CoV-2 spike RBD and block interaction with ACE2

Author

Huo, Jiangdong, Le Bas, Audrey, Ruza, Reinis R., Duyvesteyn, Helen M. E., Mikolajek, Halina, Malinauskas, Tomas, Tan, Tiong Kit, Rijal, Pramila, Dumoux, Maud, Ward, Philip N., Ren, Jingshan, Zhou, Daming, Harrison, Peter J., Weckener, Miriam, Clare, Daniel K., Vogirala, Vinod K., Radecke, Julika, Moynié, Lucile, Zhao, Yuguang, Gilbert-Jaramillo, Javier, Knight, Michael L., Tree, Julia A., Buttigieg, Karen R., Coombes, Naomi, Elmore, Michael J., Carroll, Miles W., Carrique, Loic, Shah, Pranav N. M., James, William, Townsend, Alain R., Stuart, David I., Owens, Raymond J., and Naismith, James H.

Published
2020
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37. H2S-Generating Cytosolic L-Cysteine Desulfhydrase and Mitochondrial D-Cysteine Desulfhydrase from Sweet Pepper (Capsicum annuum L.) Are Regulated During Fruit Ripening and by Nitric Oxide.

Author

Muñoz-Vargas, María A., López-Jaramillo, Javier, González-Gordo, Salvador, Paradela, Alberto, Palma, José M., and Corpas, Francisco J.

Subjects
*FRUIT ripening, *CAPSICUM annuum, *SWEET peppers, *CYSTEINE, *NITRIC oxide, *POLYACRYLAMIDE gel electrophoresis, *CLIMACTERIC, *PEPPERS, *POST-translational modification
Abstract

Aims: Pepper fruit is a horticultural product worldwide consumed that has great nutritional and economic relevance. Besides the phenotypical changes that undergo pepper fruit during ripening, there are many associated modifications at transcriptomic, proteomic, biochemical, and metabolic levels. Nitric oxide (NO) and hydrogen sulfide (H2S) are recognized signal molecules that can exert regulatory functions in diverse plant processes. This study aims at analyzing the interrelationship between NO and H2S during fruit ripening. Results: Our data indicate that the H2S-generating cytosolic L-cysteine desulfhydrase (LCD) and the mitochondrial D-cysteine desulfhydrase (DCD) activities are downregulated during ripening but this effect was reverted after NO treatment of fruits. Innovation and Conclusion: Using as a model the non-climacteric pepper fruits at different ripening stages and under an NO-enriched atmosphere, the activity of the H2S-generating LCD and DCD was analyzed. LCD and DCD activities were downregulated during ripening, but this effect was reverted after NO treatment of fruits. The analysis of LCD activity by non-denaturing polyacrylamide gel electrophoresis (PAGE) allowed identifying three isozymes designated CaLCD I to CaLCD III, which were differentially modulated by NO and strictly dependent on pyridoxal 5′-phosphate (PLP). In vitro analyses of green fruit samples in the presence of different compounds including NO donors, peroxynitrite (ONOO−), and reducing agents such as reduced glutathione (GSH) and L-cysteine (L-Cys) triggered an almost 100% inhibition of CaLCD II and CaLCD III. This redox adaptation process of both enzymes could be cataloged as a hormesis phenomenon. The protein tyrosine (Tyr) nitration (an NO-promoted post-translational modification) of the recombinant LCD was corroborated by immunoblot and by mass spectrometry (MS) analyses. Among the 11 Tyr residues present in this enzyme, MS of the recombinant LCD enabled us to identify that Tyr82 and Tyr254 were nitrated by ONOO−, this occurring near the active center on the enzyme, where His237 and Lys260 together with the cofactor PLP are involved. These data support the relationship between NO and H2S during pepper fruit ripening, since LCD and DCD are regulated by NO during this physiological event, and this could also be extrapolated to other plant species. [ABSTRACT FROM AUTHOR]

Published
2023
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40. Nitric oxide (NO) differentially modulates the ascorbate peroxidase (APX) isozymes of sweet pepper (Capsicum annuum L.) fruits

Author

Ministerio de Ciencia e Innovación (España), Zeraim Ibérica, Junta de Andalucía, González-Gordo, Salvador, Rodríguez-Ruiz, Marta, López-Jaramillo, Javier, Muñoz-Vargas, María A., Palma Martínez, José Manuel, Corpas, Francisco J., Ministerio de Ciencia e Innovación (España), Zeraim Ibérica, Junta de Andalucía, González-Gordo, Salvador, Rodríguez-Ruiz, Marta, López-Jaramillo, Javier, Muñoz-Vargas, María A., Palma Martínez, José Manuel, and Corpas, Francisco J.

Abstract

Nitric oxide (NO) is a free radical which modulates protein function and gene expression throughout all stages of plant development. Fruit ripening involves a complex scenario where drastic phenotypical and metabolic changes take place. Pepper fruits are one of the most consumed horticultural products worldwide which, at ripening, undergo crucial phenotypical and biochemical events, with NO and antioxidants being implicated. Based on previous transcriptomic (RNA‐Seq), proteomics (iTRAQ), and enzymatic data, this study aimed to identify the ascorbate peroxidase (APX) gene and protein profiles in sweet peppers and to evaluate their potential modulation by NO during fruit ripening. The data show the existence of six CaAPX genes (CaAPX1– CaAPX6) that encode corresponding APX isozymes distributed in cytosol, plastids, mitochondria, and peroxisomes. The time course expression analysis of these genes showed heterogeneous expression patterns throughout the different ripening stages, and also as a consequence of treatment with NO gas. Additionally, six APX isozymes activities (APX I–APX VI) were identified by non‐denaturing PAGE, and they were also differentially modulated during maturation and NO treatment. In vitro analyses of fruit samples in the presence of NO donors, peroxynitrite, and glutathione, showed that CaAPX activity was inhibited, thus suggesting that different posttransla-tional modifications (PTMs), including S‐nitrosation, Tyr‐nitration, and glutathionylation, respec-tively, may occur in APX isozymes. In silico analysis of the protein tertiary structure showed that residues Cys32 and Tyr235 were conserved in the six CaAPXs, and are thus likely potential targets for S‐nitrosation and nitration, respectively. These data highlight the complex mechanisms of the regulation of APX isozymes during the ripening process of sweet pepper fruits and how NO can exert fine control. This information could be useful for postharvest technology; NO regulates H2O2 levels throu

Published
2022

41. Soybean (Glycine max L.) Lipoxygenase 1 (LOX 1) Is Modulated by Nitric Oxide and Hydrogen Sulfide: An In Vitro Approach.

Author

González-Gordo, Salvador, López-Jaramillo, Javier, Palma, José M., and Corpas, Francisco J.

Subjects
*HYDROGEN sulfide, *NITRIC oxide, *FRUIT ripening, *POST-translational modification, *GERMINATION, *SOYBEAN
Abstract

Hydrogen sulfide (H2S) and nitric oxide (NO) are two relevant signal molecules that can affect protein function throughout post-translational modifications (PTMs) such as persulfidation, S-nitrosation, metal-nitrosylation, and nitration. Lipoxygenases (LOXs) are a group of non-heme iron enzymes involved in a wide range of plant physiological functions including seed germination, plant growth and development, and fruit ripening and senescence. Likewise, LOXs are also involved in the mechanisms of response to diverse environmental stresses. Using purified soybean (Glycine max L.) lipoxygenase type 1 (LOX 1) and nitrosocysteine (CysNO) and sodium hydrosulfide (NaHS) as NO and H2S donors, respectively, the present study reveals that both compounds negatively affect LOX activity, suggesting that S-nitrosation and persulfidation are involved. Mass spectrometric analysis of nitrated soybean LOX 1 using a peroxynitrite (ONOO−) donor enabled us to identify that, among the thirty-five tyrosine residues present in this enzyme, only Y214 was exclusively nitrated by ONOO−. The nitration of Y214 seems to affect its interaction with W500, a residue involved in the substrate binding site. The analysis of the structure 3PZW demonstrates the existence of several tunnels that directly communicate the surface of the protein with different internal cysteines, thus making feasible their potential persulfidation, especially C429 and C127. On the other hand, the CysNO molecule, which is hydrophilic and bulkier than H2S, can somehow be accommodated throughout the tunnel until it reaches C127, thus facilitating its nitrosation. Overall, a large number of potential persulfidation targets and the ease by which H2S can reach them through the diffuse tunneling network could be behind their efficient inhibition. [ABSTRACT FROM AUTHOR]

Published
2023
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45. A rapid antibody screening haemagglutination test for predicting immunity to SARS-CoV-2 variants of concern

Author

Ertesvåg, Nina Urke, Xiao, Julie, Zhou, Fan, Ljostveit, Sonja, Sandnes, Helene, Lartey, Sarah, Sævik, Marianne, Hansen, Lena, Madsen, Anders, Mohn, Kristin G. I., Fjelltveit, Elisabeth, Olofsson, Jan Stefan, Tan, Tiong Kit, Rijal, Pramila, Schimanski, Lisa, Øyen, Siri, Brokstad, Karl Albert, Dunachie, Susanna, Jämsén, Anni, James, William S., Harding, Adam C., Harvala, Heli, Nguyen, Dung, Roberts, David, Patel, Monika, Gopal, Robin, Zambon, Maria, Wei, Leiyan, Gilbert-Jaramillo, Javier, Knight, Michael L., Vaughan-Jackson, Alun, Dupont, Maeva, Lamikanra, Abigail A., Klennerman, Paul, Barnes, Eleanor, Deeks, Alexandra, Johnson, Sile, Skelly, Donal, Stafford, Lizzie, Townsend, Alain, Tøndel, Camilla, Kuwelker, Kanika, Blomberg, Bjørn, Bredholt, Geir, Onyango, Therese Bredholt, Vahokoski, Juha, Bansal, Amit, Trieu, Mai Chi, Amdam, Håkon, Akselsen, Per Espen, Skorge, Trude Duelien, Okkenhaug, Liv Heiberg, Linchausen, Dagrunn Waag, Langeland, Nina, and Cox, Rebecca Jane

Abstract

Background Evaluation of susceptibility to emerging SARS-CoV-2 variants of concern (VOC) requires rapid screening tests for neutralising antibodies which provide protection. Methods Firstly, we developed a receptor-binding domain-specific haemagglutination test (HAT) to Wuhan and VOC (alpha, beta, gamma and delta) and compared to pseudotype, microneutralisation and virus neutralisation assays in 835 convalescent sera. Secondly, we investigated the antibody response using the HAT after two doses of mRNA (BNT162b2) vaccination. Sera were collected at baseline, three weeks after the first and second vaccinations from older (80–99 years, n = 89) and younger adults (23–77 years, n = 310) and compared to convalescent sera from naturally infected individuals (1–89 years, n = 307). Results Here we show that HAT antibodies highly correlated with neutralising antibodies (R = 0.72–0.88) in convalescent sera. Home-dwelling older individuals have significantly lower antibodies to the Wuhan strain after one and two doses of BNT162b2 vaccine than younger adult vaccinees and naturally infected individuals. Moverover, a second vaccine dose boosts and broadens the antibody repertoire to VOC in naïve, not previously infected older and younger adults. Most (72–76%) older adults respond after two vaccinations to alpha and delta, but only 58–62% to beta and gamma, compared to 96–97% of younger vaccinees and 68–76% of infected individuals. Previously infected older individuals have, similarly to younger adults, high antibody titres after one vaccination. Conclusions Overall, HAT provides a surrogate marker for neutralising antibodies, which can be used as a simple inexpensive, rapid test. HAT can be rapidly adaptable to emerging VOC for large-scale evaluation of potentially decreasing vaccine effectiveness.

Published
2022

47. Structures and therapeutic potential of anti-RBD human monoclonal antibodies against SARS-CoV-2

Author

Huang, Kuan-Ying A., primary, Zhou, Daming, additional, Tan, Tiong Kit, additional, Chen, Charles, additional, Duyvesteyn, Helen M. E., additional, Zhao, Yuguang, additional, Ginn, Helen M., additional, Qin, Ling, additional, Rijal, Pramila, additional, Schimanski, Lisa, additional, Donat, Robert, additional, Harding, Adam, additional, Gilbert-Jaramillo, Javier, additional, James, William, additional, Tree, Julia A., additional, Buttigieg, Karen, additional, Carroll, Miles, additional, Charlton, Sue, additional, Lien, Chia-En, additional, Lin, Meei-Yun, additional, Chen, Cheng-Pin, additional, Cheng, Shu-Hsing, additional, Chen, Xiaorui, additional, Lin, Tzou-Yien, additional, Fry, Elizabeth E., additional, Ren, Jingshan, additional, Ma, Che, additional, Townsend, Alain R., additional, and Stuart, David I., additional

Published
2022
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48. T-cell and antibody responses to first BNT162b2 vaccine dose in previously infected and SARS-CoV-2-naive UK health-care workers: a multicentre prospective cohort study

Author

Angyal, Adrienn, primary, Longet, Stephanie, additional, Moore, Shona C, additional, Payne, Rebecca P, additional, Harding, Adam, additional, Tipton, Tom, additional, Rongkard, Patpong, additional, Ali, Mohammad, additional, Hering, Luisa M, additional, Meardon, Naomi, additional, Austin, James, additional, Brown, Rebecca, additional, Skelly, Donal, additional, Gillson, Natalie, additional, Dobson, Sue L, additional, Cross, Andrew, additional, Sandhar, Gurjinder, additional, Kilby, Jonathan A, additional, Tyerman, Jessica K, additional, Nicols, Alexander R, additional, Spegarova, Jarmila S, additional, Mehta, Hema, additional, Hornsby, Hailey, additional, Whitham, Rachel, additional, Conlon, Christopher P, additional, Jeffery, Katie, additional, Goulder, Philip, additional, Frater, John, additional, Dold, Christina, additional, Pace, Matthew, additional, Ogbe, Ane, additional, Brown, Helen, additional, Ansari, M Azim, additional, Adland, Emily, additional, Brown, Anthony, additional, Chand, Meera, additional, Shields, Adrian, additional, Matthews, Philippa C, additional, Hopkins, Susan, additional, Hall, Victoria, additional, James, William, additional, Rowland-Jones, Sarah L, additional, Klenerman, Paul, additional, Dunachie, Susanna, additional, Richter, Alex, additional, Duncan, Christopher J A, additional, Barnes, Eleanor, additional, Carroll, Miles, additional, Turtle, Lance, additional, de Silva, Thushan I, additional, Watson, Adam, additional, Angyal, Adrienn, additional, Alhussni, Ahmed, additional, Nicols, Alexander, additional, Deeks, Alexandra, additional, Webb-Bridges, Alice, additional, Jämsén, Anni, additional, Chawla, Anu, additional, Duncan, Christopher, additional, Conlon, Christopher, additional, O'Donnell, Denise, additional, Weeks, Esme, additional, Abuelgasim, Hibatullah, additional, Xiao, Huiyuan, additional, Spegarova, Jarmila, additional, Holmes, Jennifer, additional, Haworth, Jenny, additional, Tyerman, Jessica, additional, Kilby, Jonathan, additional, Cutteridge, Joseph, additional, Lillie, Katy, additional, Romaniuk, Leigh, additional, Denly, Lucy, additional, Hering, Luisa, additional, Ansari, M. Azim, additional, Kasanyinga, Mwila, additional, Matthews, Philippa, additional, Payne, Rebecca, additional, Wilson, Robert, additional, Rowland-Jones, Sarah, additional, Thomas, Sarah, additional, Moore, Shona, additional, Gardiner, Siobhan, additional, Tucker, Stephanie, additional, Dobson, Sue, additional, Adlou, Syed, additional, de Silva, Thushan, additional, Lawrie, Allan, additional, Smith, Nikki, additional, Turton, Helena, additional, Zawia, Amira, additional, Bayley, Martin, additional, Fairman, Alex, additional, Harrington, Kate, additional, Kirk, Rosemary, additional, Marsh, Louise, additional, Watson, Lisa, additional, Wood, Steven, additional, Diffey, Benjamin, additional, Jones, Chris, additional, Lett, Lauren, additional, Platt, Gareth, additional, Subramaniam, Krishanthi, additional, Wootton, Daniel, additional, Payne, Brendan, additional, Hambleton, Sophie, additional, Kelly, Sinead, additional, Marston, Judith, additional, Poolan, Sonia, additional, Turner, Dianne, additional, Haniffa, Muzlifah, additional, Stephenson, Emily, additional, Adele, Sandra, additional, Akhter, Hossain Delowar, additional, Chinnakannan, Senthil, additional, de Lara, Catherine, additional, Donnison, Timothy, additional, Hackstein, Carl-Philipp, additional, Lee, Lian, additional, Lim, Nicholas, additional, Malone, Tom, additional, Phillips, Eloise, additional, Ramamurthy, Narayan, additional, Robinson, Nichola, additional, Sampson, Oliver, additional, Eyre, David, additional, Simmons, Beatrice, additional, Stafford, Lizzie, additional, Mentzer, Alexander, additional, Amini, Ali, additional, Arancibia-Cárcamo, Carolina, additional, Provine, Nicholas, additional, Travis, Simon, additional, Dimitriadis, Stavros, additional, Johnson, Sile, additional, Foulkes, Sarah, additional, Khawam, Jameel, additional, Wellington, Edgar, additional, Gilbert-Jaramillo, Javier, additional, Knight, Michael, additional, Dupont, Maeva, additional, Horner, Emily, additional, Thaventhiran, James, additional, and Chalk, Jeremy, additional

Published
2022
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49. Correction: Breadth and function of antibody response to acute SARS-CoV-2 infection in humans

Author

Huang, Kuan-Ying A., primary, Tan, Tiong Kit, additional, Chen, Ting-Hua, additional, Huang, Chung-Guei, additional, Harvey, Ruth, additional, Hussain, Saira, additional, Chen, Cheng-Pin, additional, Harding, Adam, additional, Gilbert-Jaramillo, Javier, additional, Liu, Xu, additional, Knight, Michael, additional, Schimanski, Lisa, additional, Shih, Shin-Ru, additional, Lin, Yi-Chun, additional, Cheng, Chien-Yu, additional, Cheng, Shu-Hsing, additional, Huang, Yhu-Chering, additional, Lin, Tzou-Yien, additional, Jan, Jia-Tsrong, additional, Ma, Che, additional, James, William, additional, Daniels, Rodney S., additional, McCauley, John W., additional, Rijal, Pramila, additional, and Townsend, Alain R., additional

Published
2021
Full Text
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