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1.
Anal Chim Acta ; 1316: 342815, 2024 Aug 08.
Artigo em Inglês | MEDLINE | ID: mdl-38969420

RESUMO

BACKGROUND: Capillary electrophoresis (CE) has the advantage of rapid anion analysis, when employing a reverse electroosmotic flow (EOF). The conventional CE method utilizes dynamic coatings with surfactants like cetyltrimethylammonium bromide (CTAB) in the run buffer to reverse the EOF. However, this method suffers from very slow equilibration leading to drifting effective migration times of the analyte anions, which adversely affects the identification and quantification of peaks. Permanent coating of the capillary surface may obviate this problem but has been relatively little explored. Thus, permanent capillary surface modification by the covalent binding of 3-aminopropyltriethoxysilane (APTES) was studied as an alternative. RESULTS: This study investigates the effect of APTES concentration for surface functionalization on EOF mobility, separation efficiency, and reproducibility of anion separation. The performance data was complemented by X-ray photoelectron spectroscopy (XPS) and contact angle (CA) measurements. The XPS measurements showed that the coverage with APTES was dependent on its concentration in the coating solution. The XPS measurements correlated well with the EOF values determined for the capillaries tested. A standard mixture of 21 anions could be baseline separated within 10 min in the capillaries with lower EOF, but not in the capillary with the highest EOF as the residence time of the analytes was too short in this case. Compared to conventional dynamic coating with CTAB, APTES-functionalized capillaries provide faster equilibration and long-term EOF stability. The application of APTES-functionalized capillaries in analyzing different beverages demonstrates the precision, reliability, and specificity in determining organic anions, providing valuable insights of their compositions. SIGNIFICANCE: APTES coating on capillaries provides a facile approach to achieve a permanent reversal of the stable EOF to determine anions. The control of the coverage via the concentration of the reagent solution allows the tailoring of the EOF to different needs, a faster EOF for less complex samples where resolution is not challenging, while a lower EOF for higher complex samples where the focus is on separation efficiency. This enhancement in efficiency and sensitivity has been applied to analyzing organic acids in several beverages.

2.
New Phytol ; 2024 Jul 02.
Artigo em Inglês | MEDLINE | ID: mdl-38953386

RESUMO

C4 photosynthesis is a complex trait requiring multiple developmental and metabolic alterations. Despite this complexity, it has independently evolved over 60 times. However, our understanding of the transition to C4 is complicated by the fact that variation in photosynthetic type is usually segregated between species that diverged a long time ago. Here, we perform a genome-wide association study (GWAS) using the grass Alloteropsis semialata, the only known species to have C3, intermediate, and C4 accessions that recently diverged. We aimed to identify genomic regions associated with the strength of the C4 cycle (measured using δ13C), and the development of C4 leaf anatomy. Genomic regions correlated with δ13C include regulators of C4 decarboxylation enzymes (RIPK), nonphotochemical quenching (SOQ1), and the development of Kranz anatomy (SCARECROW-LIKE). Regions associated with the development of C4 leaf anatomy in the intermediate individuals contain additional leaf anatomy regulators, including those responsible for vein patterning (GSL8) and meristem determinacy (GIF1). The parallel recruitment of paralogous leaf anatomy regulators between A. semialata and other C4 lineages implies the co-option of these genes is context-dependent, which likely has implications for the engineering of the C4 trait into C3 species.

4.
J Exp Bot ; 2024 Jul 13.
Artigo em Inglês | MEDLINE | ID: mdl-39001658

RESUMO

Intercellular communication in plants, as in other multicellular organisms, allows cells in tissues to coordinate their responses for development and in response to environmental stimuli. Much of this communication is facilitated by plasmodesmata (PD), consisting of membranes and cytoplasm, that connect adjacent cells to each other. PD have long been viewed as passive conduits for the movement of a variety of metabolites and molecular cargoes, but this perception has been changing over the last two decades or so. Research from the last few years has revealed the importance of PD as signaling hubs and as crucial players in hormone signaling. The adoption of advanced biochemical approaches, molecular tools and high-resolution imaging modalities have led to several recent breakthroughs in our understanding of the roles of PD, revealing the structural and regulatory complexity of these 'protoplasmic connecting threads'. We highlight several of these findings that we think well illustrate the current understanding of PD as functioning at the nexus of plant physiology, development, and acclimation to the environment.

5.
Exp Brain Res ; 2024 Jul 04.
Artigo em Inglês | MEDLINE | ID: mdl-38963560

RESUMO

Transcranial electrical stimulation (tES) often targets the EEG-guided C3/C4 area that may not accurately represent M1 for hand muscles. This study aimed to determine if the neuroanatomy-based scalp acupuncture-guided site (AC) was a more effective spot than the C3 site for neuromodulation. Fifteen healthy subjects received one 20-minute session of high-definition transcranial alternating current stimulation (HD-tACS) intervention (20 Hz at 2 mA) at the AC or C3 sites randomly with a 1-week washout period. Subjects performed ball-squeezing exercises with the dominant hand during the HD-tACS intervention. The AC site was indiscernible from the finger flexor hotspot detected by TMS. At the baseline, the MEP amplitude from finger flexors was greater with less variability at the AC site than at the C3 site. HD-tACS intervention at the AC site significantly increased the MEP amplitude. However, no significant changes were observed after tACS was applied to the C3 site. Our results provide evidence that HD-tACS at the AC site produces better neuromodulation effects on the flexor digitorum superficialis (FDS) muscle compared to the C3 site. The AC localization approach can be used for future tES studies.

6.
J Exp Bot ; 2024 Jul 05.
Artigo em Inglês | MEDLINE | ID: mdl-38970337

RESUMO

The ratio of net CO2 uptake (Anet) and stomatal conductance (gs) is an intrinsic measurement of leaf water use efficiency (WUEi) however its measurement can be challenging for large phenotypic screens. Measurements of leaf carbon isotope composition (δ13Cleaf) may be a scalable tool to approximate WUEi for screening because it in part reflects the competing influences of Anet and gs on the CO2 partial pressure (pCO2) inside the leaf over time. However, in C4 photosynthesis the CO2 concentrating mechanism complicates the relationship between δ13Cleaf and WUEi. Despite this complicated relationship, several studies have shown genetic variation in δ13Cleaf across C4 plants. Yet there has not been a clear demonstration of whether Anet or gs are the causal mechanisms controlling WUEi and δ13Cleaf. Our approach was to characterize leaf photosynthetic traits of two Zea mays recombinant inbred lines (Z007E0067 and Z007E0150) which consistently differ for δ13Cleaf even though they have minimal confounding genetic differences. We demonstrate that these two genotypes contrasted in WUEi driven by differences in the speed of stomatal responses to changes in pCO2 and light that lead to unproductive leaf water loss. These findings provide support that differences in δ13Cleaf in closely related genotypes do reflect greater WUEi and further suggests that differences in stomatal kinetic response to changing environmental conditions is a key target to improve WUEi.

7.
Molecules ; 29(13)2024 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-38999089

RESUMO

Spin-gapless semiconductor (SGS), a class of zero-gap materials with fully spin-polarized electrons and holes, offers significant potential for high-speed, low-energy consumption applications in spintronics, electronics, and optoelectronics. Our first-principles calculations revealed that the Pca21 C4N3 monolayer exhibits a ferromagnetic ground state. Its band structure displays SGS-like characteristics, with the energy gap between the valence and conduction bands near the Fermi level in the spin-down channel much smaller than the one in the other spin channel. To enhance its SGS properties, we introduced electrons into the Pca21 C4N3 monolayer by adsorbing the CO gas molecule on its surface. Stable gas adsorption (CO@C4N3) effectively narrowed the band gap in the spin-down channel without changing the band gap in the spin-up channel obviously. Moreover, injecting holes into the CO@C4N3 system could increase the net magnetic moments and induce an SGS-to-metallic phase transition, while injecting electrons into the CO@C4N3 system is able to lower the net magnetic moments and cause an SGS-to-half-metallic phase transition. Our findings not only underscore a new promising material for practical metal-free spintronics applications but also illustrate a viable pathway for designing SGSs.

8.
Sci Rep ; 14(1): 16224, 2024 Jul 13.
Artigo em Inglês | MEDLINE | ID: mdl-39003309

RESUMO

IgA Nephropathy (IgAN) is the most prevalent glomerular disease worldwide. Complement system activation is crucial in its pathogenesis. Few studies correlated serum C3 and C4 with disease activity and prognosis. This retrospective study investigated the prognostic value of serum complement at the time of diagnosis in patients with IgAN. Specifically we evaluated whether adding serum C3 and C4 levels to established predictive models-one based on variables related to chronic kidney disease (CKD) progression and another incorporating variables from the International IgA Prediction Tool (IntIgAPT)-enhances the accuracy of outcome prediction. A composite renal outcome was defined as 50% decline in eGFR or onset of kidney failure. 101 patients were stratified according to baseline C3 levels in three groups (Low, Medium and High). During a median follow-up of 54 months, the Low group exhibited higher incidence of primary outcome (16.3 events vs 2.9 and 1.7 events × 100 pts/year, p = 0.0026). Model-1 (M1), consisting of CKD progression variables, and Model-3 (M3), comprising IntIgANPT variables, were implemented with baseline C3 and C4 to create Model-2 (M2) and Model-4 (M4), respectively. M2 demonstrated better predictive performance over M1, showing higher discrimination (lower AIC and BIC, higher C-index and NR2). Similarly, M4 outperformed M3, showing enhanced outcome prediction when C3 and C4 levels were added. Implementation of serum C3 and C4 can enhance prediction accuracy of already-validated prognostic models in IgAN. Lower C3 and higher C4 levels were associated with poorer prognosis, highlighting a more 'Complement-Pathic' subset of patients.


Assuntos
Complemento C3 , Complemento C4 , Glomerulonefrite por IGA , Humanos , Glomerulonefrite por IGA/sangue , Masculino , Feminino , Complemento C3/metabolismo , Complemento C3/análise , Adulto , Complemento C4/metabolismo , Complemento C4/análise , Estudos Retrospectivos , Prognóstico , Pessoa de Meia-Idade , Progressão da Doença , Taxa de Filtração Glomerular , Insuficiência Renal Crônica/sangue , Biomarcadores/sangue , Rim/patologia , Rim/fisiopatologia
9.
BMC Genomics ; 25(1): 688, 2024 Jul 13.
Artigo em Inglês | MEDLINE | ID: mdl-39003461

RESUMO

BACKGROUND: The co-occurrence of C4 and CAM photosynthesis in a single species seems to be unusual and rare. This is likely due to the difficulty in effectively co-regulating both pathways. Here, we conducted a comparative transcriptomic analysis of leaves and cotyledons of the C4-like species Sesuvium sesuvioides (Aizoaceae) using RNA-seq. RESULTS: When compared to cotyledons, phosphoenolpyruvate carboxylase 4 (PEPC4) and some key C4 genes were found to be up-regulated in leaves. During the day, the expression of NADP-dependent malic enzyme (NADP-ME) was significantly higher in cotyledons than in leaves. The titratable acidity confirmed higher acidity in the morning than in the previous evening indicating the induction of weak CAM in cotyledons by environmental conditions. Comparison of the leaves of S. sesuvioides (C4-like) and S. portulacastrum (C3) revealed that PEPC1 was significantly higher in S. sesuvioides, while PEPC3 and PEPC4 were up-regulated in S. portulacastrum. Finally, potential key regulatory elements involved in the C4-like and CAM pathways were identified. CONCLUSIONS: These findings provide a new species in which C4-like and CAM co-occur and raise the question if this phenomenon is indeed so rare or just hard to detect and probably more common in succulent C4 lineages.


Assuntos
Aizoaceae , Cotilédone , Perfilação da Expressão Gênica , Fotossíntese , Folhas de Planta , Cotilédone/genética , Cotilédone/metabolismo , Folhas de Planta/metabolismo , Folhas de Planta/genética , Fotossíntese/genética , Aizoaceae/genética , Aizoaceae/metabolismo , Regulação da Expressão Gênica de Plantas , Transcriptoma , Malato Desidrogenase/metabolismo , Malato Desidrogenase/genética , Fosfoenolpiruvato Carboxilase/metabolismo , Fosfoenolpiruvato Carboxilase/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo
10.
J Exp Clin Cancer Res ; 43(1): 194, 2024 Jul 17.
Artigo em Inglês | MEDLINE | ID: mdl-39014480

RESUMO

Continued exploration of the androgen receptor (AR) is crucial, as it plays pivotal roles in diverse diseases such as prostate cancer (PCa), serving as a significant therapeutic focus. Therefore, the Department of Urology Dresden hosted an international meeting for scientists and clinical oncologists to discuss the newest advances in AR research. The 2nd International Androgen Receptor Symposium was held in Dresden, Saxony, Germany, from 26-27.04.2024, organised by Dr. Holger H.H. Erb. Following the format of the first meeting, more than 35 scientists from 8 countries attended the event to discuss recent developments, research challenges, and identification of venues in AR research. An important new feature was the involvement of PhD students and young investigators, acknowledging the high scientific quality of their work. The symposium included three covers: new advances from clinical research, basic and translational research, and novel strategies to target AR. Moreover, based on its increasing clinical relevance, a PSMA theranostic mini-symposium was added at the end of the AR symposium to allow the audience to discuss the newest advances in PSMA theranostic. This report focuses on the highlights and discussions of the meeting.


Assuntos
Neoplasias da Próstata , Receptores Androgênicos , Humanos , Masculino , Neoplasias da Próstata/metabolismo , Neoplasias da Próstata/terapia , Neoplasias da Próstata/genética , Receptores Androgênicos/metabolismo , Receptores Androgênicos/genética
11.
Materials (Basel) ; 17(12)2024 Jun 20.
Artigo em Inglês | MEDLINE | ID: mdl-38930403

RESUMO

Alite(C3S)-Ye'elimite(C4A3$) cement is a high cementitious material that incorporates a precise proportion of ye'elimite into the ordinary Portland cement. The synthesis and hydration behavior of Alite-Ye'elimite clinker with different lime saturation factors were investigated. The clinkers were synthesized using a secondary thermal treatment process, and their compositions were characterized. The hydrated pastes were analyzed for their hydration products, pore structure, mechanical strength, and microstructure. The clinkers and hydration products were characterized using XRD, TG-DSC, SEM, and MIP analysis. The results showed that the Alite-Ye'elimite cement clinker with a lime saturation factor (KH) of 0.93, prepared through secondary heat treatment, contained 64.88% C3S and 2.06% C4A3$. At this composition, the Alite-Ye'elimite cement clinker demonstrated the highest 28-day strength. The addition of SO3 to the clinkers decreased the content of tricalcium aluminate (C3A) and the ratio of Alite/Belite (C3S/C2S), resulting in a preference for belite formation. The pore structure of the hydrated pastes was also investigated, revealing a distribution of pore sizes ranging from 0.01 to 10 µm, with two peaks on each differential distribution curve corresponding to micron and sub-micron pores. The pore volume decreased from 0.22 ± 0.03 to 0.15 ± 0.18 cm3 g-1, and the main peak of pore distribution shifted towards smaller sizes with increasing hydration time.

12.
Immunol Allergy Clin North Am ; 44(3): 543-560, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-38937015

RESUMO

The role of contact system activation has been clearly established in the pathogenesis of hereditary angioedema due to C1 inhibitor deficiency (HAE-C1INH). C1 inhibitor (C1INH)-protease complexes, levels of functional C1INH, plasma kallikrein activation, and cleavage of high-molecular-weight kininogen have each been associated with disease activity. More recently, HAE with normal levels of C1INH (HAE-nl-C1INH) has been recognized. Six genetic mutations have been identified which are linked to HAE-nl-C1INH phenotypes. The majority of individuals with HAE-nl-C1INH fall into the unknown category. There is substantial evidence that bradykinin generation underlies the recurrent attacks of swelling in some of these cohorts.


Assuntos
Biomarcadores , Bradicinina , Proteína Inibidora do Complemento C1 , Humanos , Bradicinina/metabolismo , Proteína Inibidora do Complemento C1/metabolismo , Angioedema/diagnóstico , Angioedema/metabolismo , Angioedema/etiologia , Angioedemas Hereditários/diagnóstico , Angioedemas Hereditários/metabolismo , Angioedemas Hereditários/etiologia , Mutação
13.
Sci Rep ; 14(1): 14778, 2024 Jun 26.
Artigo em Inglês | MEDLINE | ID: mdl-38926406

RESUMO

Optimal pH is essential for efficient cobalt extraction from polymeric membrane systems, with D2EHPA used as an extractant for Co(II) at pH < 7, achieving 47% efficiency. The pH of piperazine as a stripping agent increases to a concentration of 0.48 M, and the extraction efficiency of Co(II) > 80%. Depending on the functional group of (C4H10N2), the optimal pH for separation was 9.8. The study revealed that pKa value was calculated to predict the ideal pH, and its value was 9.73, which is nearly to the pH, since the pH of the strip concentration and the properties of the membrane affect the extraction of cobalt at 30 °C. The partition ratio indicates the high distribution of the extract in supported ceramic polymer membrane (SCPM). The ceramic component provides mechanical strength and rigidity to the overall membrane structure, allowing it to withstand high pressures and temperatures during operation Study various factors such as the effect of pH on the ionization of the extract; effect of pH on band ionization; effect of pH on the temperature in the extract, effect of pH on the solute, effect of the band at different pH ranges and a comparison was made between the predictive model and experimental data that was proven through mathematical modeling using the MATLAB program.

14.
Bioresour Technol ; 405: 130932, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-38838831

RESUMO

The first comparative pre-treatment study of Miscanthus (Mxg) and sugarcane bagasse (SCB) using steam explosion (SE) and pressurised disc refining (PDR) pretreatment to optimise xylose and xylo-oligosaccharide release is described. The current investigation aimed to 1) Develop optimised batch-wise steam explosion parameters for Mxg and SCB, 2) Scale from static batch steam explosion to dynamic continuous pressurised disc refining, 3) Identify, understand, and circumvent scale-up production hurdles. Optimised SE parameters released 82% (Mxg) and 100% (SCB) of the available xylan. Scaling to PDR, Miscanthus yielded 85% xylan, highlighting how robust scouting assessments for boundary process parameters can result in successful technical transfer. In contrast, SCB technical transfer was not straightforward, with significant differences observed between the two processes, 100% (SE) and 58% (PDR). This report underlines the importance of feedstock-specific pretreatment strategies to underpin process development, scale-up, and optimisation of carbohydrate release from biomass.


Assuntos
Celulose , Oligossacarídeos , Poaceae , Saccharum , Vapor , Xilose , Saccharum/química , Celulose/química , Projetos Piloto , Biotecnologia/métodos , Xilanos , Glucuronatos
15.
Nefrologia (Engl Ed) ; 44(3): 402-407, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38906767

RESUMO

BACKGROUND: There is a little information about of expression of C4d (complement fragment) in Focal segmental glomerulosclerosis (FSGS) subtypes. Our aim was to determine the expression of C4d in FSGS subtypes in percutaneous native renal biopsies in a second-level hospital and its correlation with clinical, biochemical and histological variables. MATERIAL AND METHODS: A retrospective study in paraffin blocks of patients with biopsy with FSGS aged 16-65 years, indistinct sex, not diabetic or obese. Immunohistochemistry was performed for C4d and their expression was analyzing in non-sclerosed glomerular capillaries (GC) and sclerosis areas (SA). Clinical and biochemical variables were recorded. The cases were divided into C4d positive and C4d negative groups and compared. The correlation between C4d staining scores in CG and SA with clinical and biochemical variables were analyzed. RESULTS: Twenty samples were analyzed, 4 for each subtype. At the time of biopsy average age 38.8 ±â€¯18.6 years, 65% male, 8.7% were hypertension. The percentage of positivity for C4d was 40% in GC, 30% SA and 35% in mesangium. The highest expression was for cellular and collapsing subtypes. C4d positivity cases had increased proteinuria (p = 0.035). A significant correlation was found between percentage of C4d expression in CG with SA (p = 0.012) and SA with tubular atrophy and interstitial fibrosis (p < 0.05). CONCLUSIONS: C4d expression in FSGS predominated in the cellular and collapsing subtypes, which translates complement activation. C4d is a possible surrogate marker in GSFS.


Assuntos
Complemento C4b , Glomerulosclerose Segmentar e Focal , Humanos , Masculino , Glomerulosclerose Segmentar e Focal/patologia , Adulto , Feminino , Estudos Retrospectivos , Pessoa de Meia-Idade , Adolescente , Adulto Jovem , Idoso , Complemento C4b/análise , Fragmentos de Peptídeos/análise
16.
Medicina (B Aires) ; 84(3): 468-473, 2024.
Artigo em Espanhol | MEDLINE | ID: mdl-38907960

RESUMO

INTRODUCTION: Membranous nephropathy (MN) is the most common cause of primary nephrotic syndrome in adults (20-30%). Light microscopy shows thickening of glomerular basement membrane with appearance of spikes. These histological findings are not evident in early forms, in which case the granular deposition pattern of IgG and/or C3 in the basement membrane by immunofluorescence (IF) constitutes the diagnostic tool that allows to differentiate it from minimal change disease (MCD). Complement system plays a key role in the pathophysiology of MN. C4d is a degradation product and a marker of the complement system activation. C4d labelling by immunohistochemical (HI) technique can help in the differential diagnosis between both glomerulopathies NM and MCD when the material for IF is insufficient and light microscopy is normal. Our objective was to explore the discrimination power of C4d to differentiate between MN and MCD in renal biopsy material. METHODS: Paraffin-embedded samples were recovered from renal biopsies with a diagnosis of MN and MCD performed between 1/1/2008 and 4/1/2019. IH staining was performed by immunoperoxidase technique using a rabbit anti-human C4d polyclonal antibody. RESULTS: In all cases with MN (n = 27, 15 males) with a median age of 63 (range: 18-87) years, C4d deposits were detected. In 21 cases with MCD (12 males) with a median age of 51 (range: 18-87) years, the C4d marking was negative in every samples. CONCLUSION: The results indicate that the marking of the renal biopsy with C4d is a useful tool for the differential diagnosis between NM and MCD.


Introducción: La nefropatía membranosa (NM) es la causa más frecuente de síndrome nefrótico primario en adultos (20-30%). En la microscopia óptica se observa engrosamiento de membrana basal glomerular con aparición de espigas. Estos hallazgos histológicos no son evidentes en formas tempranas, en cuyo caso el patrón de depósito granular de IgG y/o C3 en la membrana basal por inmunofluorescencia (IF) permite diferenciarla de enfermedad por cambios mínimos (ECM). El sistema del complemento juega un papel central en la fisiopatología de la NM. C4d es producto de degradación y un marcador de la activación del complemento. La marcación con C4d en muestras de biopsias renales, por técnica de inmunohistoquímica (IH) puede colaborar en el diagnóstico diferencial entre ambas glomerulopatías. Nuestro objetivo fue explorar el poder de discriminación del C4d para diferenciar NM de ECM en material de biopsias renales. Métodos: Se recuperaron muestras en parafina de biopsias renales con diagnóstico de NM y ECM realizados entre 1/1/2008 y 1/4/2019. Se realizaron tinciones de IH por técnica de inmunoperoxidasa con C4d usando un anticuerpo policlonal antihumano de conejo. Resultados: En todos los casos con NM (n = 27, 15 hombres) con mediana de edad de 63 (rango: 18-86) años se detectaron depósitos de C4d. En los 21 casos con ECM (12 hombres) con mediana de edad de 51 (rango: 18-87) años la marcación de C4d fue negativa. Conclusión: Los resultados indican que la marcación de la biopsia renal con C4d es una herramienta útil para el diagnóstico diferencial entre NM y ECM.


Assuntos
Complemento C4b , Glomerulonefrite Membranosa , Glomerulonefrite Membranosa/diagnóstico , Glomerulonefrite Membranosa/patologia , Glomerulonefrite Membranosa/imunologia , Humanos , Masculino , Adulto , Pessoa de Meia-Idade , Feminino , Idoso , Complemento C4b/análise , Adulto Jovem , Diagnóstico Diferencial , Idoso de 80 Anos ou mais , Adolescente , Biópsia , Biomarcadores/análise , Nefrose Lipoide/patologia , Nefrose Lipoide/diagnóstico , Fragmentos de Peptídeos/análise , Estudos Retrospectivos
17.
J Biol Chem ; 300(7): 107478, 2024 Jun 13.
Artigo em Inglês | MEDLINE | ID: mdl-38879009

RESUMO

Antigenically sequence variable M proteins of the major bacterial pathogen Streptococcus pyogenes (Strep A) are responsible for recruiting human C4b-binding protein (C4BP) to the bacterial surface, which enables Strep A to evade destruction by the immune system. The most sequence divergent portion of M proteins, the hypervariable region (HVR), is responsible for binding C4BP. Structural evidence points to the conservation of two C4BP-binding sequence patterns (M2 and M22) in the HVR of numerous M proteins, with this conservation applicable to vaccine immunogen design. These two patterns, however, only partially explain C4BP binding by Strep A. Here, we identified several M proteins that lack these patterns but still bind C4BP and determined the structures of two, M68 and M87 HVRs, in complex with a C4BP fragment. Mutagenesis of these M proteins led to the identification of amino acids that are crucial for C4BP binding, enabling formulation of new C4BP-binding patterns. Mutagenesis was also carried out on M2 and M22 proteins to refine or generate experimentally grounded C4BP-binding patterns. The M22 pattern was the most prevalent among M proteins, followed by the M87 and M2 patterns, while the M68 pattern was rare. These patterns, except for M68, were also evident in numerous M-like Enn proteins. Binding of C4BP via these patterns to Enn proteins was verified. We conclude that C4BP-binding patterns occur frequently in Strep A strains of differing M types, being present in their M or Enn proteins, or frequently both, providing further impetus for their use as vaccine immunogens.

19.
Plants (Basel) ; 13(9)2024 Apr 25.
Artigo em Inglês | MEDLINE | ID: mdl-38732417

RESUMO

This study aimed to investigate the impact of cold stress and priming on photosynthesis in the early development of maize and soybean, crops with diverse photosynthetic pathways. The main objectives were to determine the effect of cold stress on chlorophyll a fluorescence parameters and spectral reflectance indices, to determine the effect of cold stress priming and possible stress memory and to determine the relationship between different parameters used in determining the stress response. Fourteen maize inbred lines and twelve soybean cultivars were subjected to control, cold stress, and priming followed by cold stress in a walk-in growth chamber. Measurements were conducted using a portable fluorometer and a handheld reflectance instrument. Cold stress induced an overall downregulation of PSII-related specific energy fluxes and efficiencies, the inactivation of RCs resulting in higher energy dissipation, and electron transport chain impairment in both crops. Spectral reflectance indices suggested cold stress resulted in pigment differences between crops. The effect of priming was more pronounced in maize than in soybean with mostly a cumulatively negative effect. However, priming stabilized the electron trapping efficiency and upregulated the electron transfer system in maize, indicating an adaptive response. Overall, this comprehensive analysis provides insights into the complex physiological responses of maize and soybean to cold stress, emphasizing the need for further genotype-specific cold stress response and priming effect research.

20.
Front Bioeng Biotechnol ; 12: 1398467, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38812916

RESUMO

Acetogens are among the key microorganisms involved in the bioproduction of commodity chemicals from diverse carbon resources, such as biomass and waste gas. Thermophilic acetogens are particularly attractive because fermentation at higher temperatures offers multiple advantages. However, the main target product is acetic acid. Therefore, it is necessary to reshape metabolism using genetic engineering to produce the desired chemicals with varied carbon lengths. Although such metabolic engineering has been hampered by the difficulty involved in genetic modification, a model thermophilic acetogen, M. thermoacetica ATCC 39073, is the case with a few successful cases of C2 and C3 compound production, other than acetate. This brief report attempts to expand the product spectrum to include C4 compounds by using strain Y72 of Moorella thermoacetica. Strain Y72 is a strain related to the type strain ATCC 39073 and has been reported to have a less stringent restriction-modification system, which could alleviate the cumbersome transformation process. A simplified procedure successfully introduced a key enzyme for acetoin (a C4 chemical) production, and the resulting strains produced acetoin from sugars and gaseous substrates. The culture profile revealed varied acetoin yields depending on the type of substrate and culture conditions, implying the need for further engineering in the future. Thus, the use of a user-friendly chassis could benefit the genetic engineering of M. thermoacetica.

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