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1.
Asian Spine J ; 18(2): 260-264, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38650091

RESUMO

STUDY DESIGN: A retrospective, cross-sectional study was conducted to analyze the implications of asymmetric baseball movements on the incidence of spondylolysis. PURPOSE: This study aimed to evaluate the relationship between asymmetric movements and the laterality of spondylolysis. OVERVIEW OF LITERATURE: Baseball, characterized by its asymmetric throwing and batting, may disproportionately stress one side. Lumbar spondylolysis is a frequent cause of lower back pain in young athletes, particularly those involved in activities with consistent unilateral rotations such as baseball. However, whether a link exists between the laterality in spondylolysis and the dominant throwing/ batting side or whether disparities exist between pitchers and fielders remains unclear. METHODS: The study included 85 players. Participants were divided into two groups: pitchers and fielders. The association between the laterality of spondylolysis and the throwing/batting side in the overall cohort and between the two groups was evaluated. RESULTS: Among pitchers, 16 lesions appeared on the throwing side and 32 on the nonthrowing side (p =0.029). For fielders, no notable difference was observed between the two sides (p =0.363). Furthermore, batting preference did not influence the laterality of spondylolysis in either group. CONCLUSIONS: Adolescent baseball players, particularly pitchers, exhibited a higher incidence of lumbar spondylolysis on the side opposite their throwing arm. The findings of this study highlight the significant effect of asymmetrical sporting activities on the development of spondylolysis, to which pitchers are particularly susceptible.

2.
Int J Biol Macromol ; 261(Pt 1): 129724, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38272403

RESUMO

Proteinaceous liquid droplets, generated by liquid-liquid phase separation, function as membraneless compartments that are essential for diverse biological functions. Studies addressing droplet generation have used 1,6-hexanediol (1,6-HD) as a droplet-discerning agent owing to its capacity to induce droplet deformation. Despite the empirical utility of 1,6-HD, the mechanism underlying 1,6-HD-induced droplet deformation remains unknown. In this study, the solubilities of N-acetyl amino acid amides, which correspond to proteinogenic amino acid residues, were examined in the presence of 1,6-HD at 25 °C. Other solvents included ethanol, 1-propanol, and amides. Remarkably, 1,6-HD effectively solubilized hydrophobic species (particularly aromatic species) and exhibited reduced efficacy in solubilizing hydrophilic species and peptide bond moieties. These solubilizing effects are reflected in changes in protein solubility and structure. Specifically, 1,6-HD primarily targets the hydrophobic regions of a protein, increasing protein solubility without causing substantial structural changes. This solubilization mechanism is essential for elucidating the role of 1,6-HD as a droplet-discerning agent and recognizing its potential limitations.


Assuntos
Amidas , Aminoácidos , Glicóis , Solubilidade , Amidas/química , Solventes/química , Água , Proteínas
3.
Nano Lett ; 23(23): 11167-11173, 2023 Dec 13.
Artigo em Inglês | MEDLINE | ID: mdl-37987714

RESUMO

Carbon nanobelts (CNBs) correspond to carbon nanotube (CNT) segments and are insoluble in most common aqueous solutions, posing challenges across diverse applications. In this study, [12] CNB, which corresponds to a (6,6) CNT segment, was solubilized by aliphatic surfactant micelles through host-guest complexation, which was confirmed by comprehensive analyses involving spectrophotometry, mass spectrometry, and molecular dynamics simulations. Through this solubilization, zero-Stokes shift emission of the CNB could occur, which could be ascribed to the symmetry-allowed transition. In contrast, CNB was insoluble in non-aliphatic surfactant solutions. The mechanism by which CNB is solubilized using aliphatic surfactants is completely distinct from that of the CNT dispersion mechanism. The present finding provides knowledge of the effectiveness of aliphatic compounds in solubilizing CNBs and their derivatives (carbon nanohoops), which show significant potential for various applications in aqueous systems, including biological applications.

4.
BMC Musculoskelet Disord ; 24(1): 558, 2023 Jul 08.
Artigo em Inglês | MEDLINE | ID: mdl-37422627

RESUMO

BACKGROUND: Lumbar spondylolysis, a common identifiable cause of low back pain in young athletes, reportedly has a higher incidence rate in males. However, the reason for its higher incidence in males is not clear. This study aimed to investigate the epidemiological differences between the sexes in adolescent patients with lumbar spondylolysis. METHODS: A retrospective study was conducted in 197 males and 64 females diagnosed with lumbar spondylolysis. These patients visited our institution from April 2014 to March 2020 with their main complaint being low back pain, and they were followed-up until the end of their treatment. We investigated associations between lumbar spondylosis, their background factors, and characteristics of the lesions and analyzed their treatment results. RESULTS: Males had a higher prevalence of spina bifida occulta (SBO) (p = 0.0026), more lesions with bone marrow edema (p = 0.0097), and more lesions in the L5 vertebrae (p = 0.021) than females. The popular sports disciplines were baseball, soccer, and track and field in males, and volleyball, basketball, softball in females. The dropout rate, age at diagnosis, bone union rate, and treatment period did not differ between the sexes. CONCLUSION: Lumbar spondylolysis was more common in males than in females. SBO, bone marrow edema, and L5 lesions were more frequent in males, and sports discipline varied between the sexes.


Assuntos
Basquetebol , Dor Lombar , Espinha Bífida Oculta , Espondilólise , Masculino , Feminino , Humanos , Adolescente , Dor Lombar/etiologia , Japão/epidemiologia , Estudos Retrospectivos , Espondilólise/epidemiologia , Vértebras Lombares/patologia , Espinha Bífida Oculta/complicações , Espinha Bífida Oculta/epidemiologia , Espinha Bífida Oculta/patologia
5.
J Chromatogr A ; 1703: 464112, 2023 Aug 16.
Artigo em Inglês | MEDLINE | ID: mdl-37285623

RESUMO

Immobilized metal ion affinity chromatography (IMAC) is one of the most common purification techniques for histidine-tagged proteins (His-tagged proteins). IMAC enables the purification of His-tagged proteins at high purity on the basis of coordination bonds between His-tags and metal ions (such as Ni2+, Co2+, and Cu2+) immobilized on the matrices in columns. However, IMAC requires low-pH solutions or high-concentration imidazole solutions for eluting His-tagged proteins, which can affect protein conformation and activity. The present study provides a His-tagged protein purification method using zirconia particles modified with phosphate groups. This method is based on the electrostatic attractions between a His-tag moiety of proteins and phosphate groups on the zirconia particles; this method requires only high-concentration salt solutions at pH 7.0 for eluting the proteins. A column packed with phosphate-modified zirconia particles was demonstrated to enable the purification of two model His-tagged proteins-His-tagged green fluorescent protein and His-tagged alkaline phosphatase fused with maltose binding protein. Thus, this chromatography method is useful for purifying His-tagged proteins without any pH stress or additives. Additionally, because of the mechanical properties of the zirconia particles, this technique enables high-performance purification at a high flow rate.


Assuntos
Histidina , Fosfatos , Histidina/química , Indicadores e Reagentes , Cromatografia de Afinidade/métodos , Proteínas Recombinantes
6.
J Med Case Rep ; 17(1): 253, 2023 Jun 11.
Artigo em Inglês | MEDLINE | ID: mdl-37301873

RESUMO

BACKGROUND: Various neurological manifestations associated with coronavirus disease 2019 have been increasingly reported. Herein, we report a rare case of anterior interosseous nerve syndrome, which occurred 5 days after the onset of coronavirus disease 2019. CASE PRESENTATION: A 62-year-old Asian woman with a history of coronavirus disease 2019 who developed a complete motor deficit in the left flexor pollicis longus and pronator quadratus without sensory deficits. The symptoms appeared as a sudden onset fatigue and severe pain of the left arm, 5 days after the onset of coronavirus disease 2019. She noticed paralysis of the left thumb at 2 weeks after the onset of coronavirus disease 2019. Electromyography assessment of the anterior interosseous nerve-dominated muscles revealed neurogenic changes such as positive sharp wave and fibrillation in flexor pollicis longus and pronator quadratus, confirming the diagnosis of anterior interosseous nerve syndrome. There were no other diseases that could have resulted in peripheral nerve palsy. We performed a functional reconstruction surgery of the thumb by tendon transfer from the extensor carpi radialis longus to the flexor pollicis longus. The patient reported a good patient-reported outcome (2.27 points in QuickDASH Disability/Symptom scoring and 5 points in Hand20 scoring) at final follow-up (1 year after the surgery). CONCLUSION: This case highlights the need for vigilance regarding the possible development of anterior interosseous nerve syndrome in patients with coronavirus disease 2019. Tendon transfer from extensor carpi radialis longus to flexor pollicis longus can provide good functional recovery for unrecovered motor paralysis after anterior interosseous nerve syndrome.


Assuntos
COVID-19 , Feminino , Humanos , Pessoa de Meia-Idade , COVID-19/complicações , Polegar/inervação , Nervo Mediano , Músculo Esquelético , Paralisia/etiologia
7.
Biophys Physicobiol ; 20(1): e200002, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37234848
8.
Int J Mol Sci ; 24(7)2023 Apr 03.
Artigo em Inglês | MEDLINE | ID: mdl-37047640

RESUMO

The present study aimed to evaluate the effects of Botryococcus terribilis ethanol extract (BTEE) on lipopolysaccharide (LPS)-induced inflammation in RAW264 cells. BTEE significantly attenuated LPS-induced nitric oxide production and inflammatory cytokines release, including Ccl2, Cox2, and Il6. On the other hand, several anti-inflammatory mediators, such as Pgc1ß and Socs1, were increased in BTEE-treated cells. Further, we performed an untargeted whole-genome microarray analysis to explore the anti-inflammatory molecular mechanism of BTEE. Enrichment analysis showed BTEE significantly downregulated 'response to stimulus', 'locomotion', and 'immune system response' and upregulated 'cell cycle' gene ontologies in both 6- and 17-h post-LPS stimulation conditions. Pathway analysis revealed BTEE could downregulate the expressions of chemokines of the CC and CXC subfamily, and cytokines of the TNF family, TGFß family, IL1-like, and class I helical. PPI analysis showed AXL receptor tyrosine kinase (Axl), a receptor tyrosine kinase from the TAM family, and its upstream transcription factors were downregulated in both conditions. Node neighborhood analysis showed several Axl coexpressed genes were also downregulated. Further, kinase enrichment and chemical perturbation analyses supported Axl inhibition in BTEE-treated conditions. Altogether, these findings suggest anti-inflammatory effects of BTEE that are mediated via the suppression of pro-inflammatory cytokines and predict its potential as an Axl inhibitor.


Assuntos
Etanol , Macrófagos , Animais , Camundongos , Etanol/farmacologia , Macrófagos/metabolismo , Lipopolissacarídeos/farmacologia , Anti-Inflamatórios/farmacologia , Anti-Inflamatórios/metabolismo , Células RAW 264.7 , Citocinas/metabolismo , Óxido Nítrico/metabolismo , NF-kappa B/metabolismo , Óxido Nítrico Sintase Tipo II/metabolismo
9.
Colloids Surf B Biointerfaces ; 226: 113291, 2023 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-37031515

RESUMO

High-performance and cost-effective purification is necessary for the development of antibody drugs. This study found that nanoparticles of zirconia modified with phosphate groups selectively adsorb immunoglobulin G (IgG) antibodies against serum proteins with high adsorption capacity. The IgG antibodies collected from the zirconia nanoparticle surfaces retain their molecular conformation. Importantly, zirconia nanoparticles have the highest affinity for human IgG antibodies among tested mammalian IgG antibodies. The affinity for human IgG subclasses is in the order IgG3 > IgG1 > IgG2, which contrasts with a conventional ligand (Protein A) that has a lower affinity for IgG3. Because zirconia nanoparticles are chemically and mechanically stable, they can be utilized for the purification of antibody drugs not only in batch methods but also in chromatography as a process upstream or downstream of Protein A chromatography and even as an alternative process.


Assuntos
Imunoglobulina G , Fosfatos , Animais , Humanos , Adsorção , Mamíferos
10.
ACS Med Chem Lett ; 14(4): 396-404, 2023 Apr 13.
Artigo em Inglês | MEDLINE | ID: mdl-37077386

RESUMO

Deregulating fibroblast growth factor receptor (FGFR) signaling is a promising strategy for cancer therapy. Herein, we report the discovery of compound 5 (TAS-120, futibatinib), a potent and selective covalent inhibitor of FGFR1-4, starting from a unique dual inhibitor of mutant epidermal growth factor receptor and FGFR (compound 1). Compound 5 inhibited all four families of FGFRs in the single-digit nanomolar range and showed high selectivity for over 387 kinases. Binding site analysis revealed that compound 5 covalently bound to the cysteine 491 highly flexible glycine-rich loop region of the FGFR2 adenosine triphosphate pocket. Futibatinib is currently in Phase I-III trials for patients with oncogenically driven FGFR genomic aberrations. In September 2022, the U.S. Food & Drug Administration granted accelerated approval for futibatinib in the treatment of previously treated, unresectable, locally advanced, or metastatic intrahepatic cholangiocarcinoma harboring an FGFR2 gene fusion or other rearrangement.

11.
J Pharm Health Care Sci ; 9(1): 9, 2023 Mar 02.
Artigo em Inglês | MEDLINE | ID: mdl-36859482

RESUMO

BACKGROUND: Supporting people living with HIV using anti-retroviral therapy (ART) is important due to the requirement for strict medication adherence. To date, no data from longitudinal studies evaluating adherence by treatment-naïve people living with HIV are currently available. We investigated the adherence of treatment-naïve people living with HIV over time and examined the relationships among decisional conflicts, adherence, and health-related quality of life (HRQL). METHODS: The survey items included adherence (visual analogue scale [VAS]), decisional conflict (decisional conflict scale [DCS]), and HRQL (Medical Outcomes Study HIV Health Survey [MOS-HIV]). The DCS and MOS-HIV scores and the VAS and MOS scores were collected electronically at the ART initiation time point and at 4-, 24-, and 48-week post-treatment time points. RESULTS: A total of 215 participants were enrolled. The mean DCS score was 27.3 (SD, 0.9); 23.3% of participants were in the high-score and 36.7% in the low-score groups. The mean adherence rates at 4, 24, and 48 weeks were 99.2% (standard error [SE], 0.2), 98.4% (SE, 0.4), and 96.0% (SE, 1.2), respectively. The least-square means of the MOS-HIV for the DCS (high vs. low scores) were 64.4 vs. 69.2 for general health perceptions and 57.7 vs. 64.0 for HRQL, respectively. CONCLUSION: Adherence among treatment-naïve people living with HIV was maintained at a higher level, and HRQL tended to improve with ART. People with high levels of decisional conflict tended to have lower HRQL scores. Support for people living with HIV during ART initiation may be related to HRQL.

12.
Spine Surg Relat Res ; 7(1): 83-88, 2023 Jan 27.
Artigo em Inglês | MEDLINE | ID: mdl-36819633

RESUMO

Introduction: Lumbar spondylolysis typically arises during adolescence, whereas its onset in adulthood is rare. Several studies have reported incidentally identified terminal-stage spondylolysis in adults, but only one case series has investigated acute lumbar spondylolysis lesions with bone marrow edema in adults. Methods: We retrospectively investigated lumbar spondylolysis in patients aged 18 years or older. Age at diagnosis, sex, competitive sporting level, and competitive sporting discipline were investigated in each patient. The level of the affected vertebra, pathological stage, bone union, and treatment period were analyzed for each lesion. Results: The study included nine patients (eight males and one female), aged 18-22 years old, with 14 acute lumbar spondylolysis lesions. Four patients were soccer players (two professionals, and two amateurs), four were amateur track and field athletes, and one was an amateur basketball player. The affected vertebral levels were L3 in two lesions, L4 in seven lesions, and L5 in five lesions. The pathological stage was pre-lysis stage in two lesions, early stage in eight lesions, and progressive stage in four lesions. Ten lesions achieved bone union with conservative therapy. The average treatment period was 84.7 days. Conclusions: The adult-onset lumbar spondylolysis patients were young, up to their early 20s, and generally professional sportspeople performing at an elite level. Most of them were performing a sport that has been reported to have a high risk of causing lumbar spondylolysis. L4 was the most affected vertebral level in contrast to L5, which has been reported in adolescent lumbar spondylolysis. Bone union was achieved for most lesions with conservative therapy.

13.
Nanoscale ; 15(5): 2340-2353, 2023 Feb 02.
Artigo em Inglês | MEDLINE | ID: mdl-36637062

RESUMO

Carbon nanotubes (CNTs) have adverse impacts on metabolism in biological systems. The impacts should be associated with interactions of the CNTs with coenzymes, such as nicotinamide adenine dinucleotide (NAD), because most metabolic processes are governed by coenzyme-dependent reactions. This study demonstrates that NAD molecules adsorb onto the CNT surface, leading to the formation of interfacial NAD layers-in other words, a coenzyme corona (coenzyme-based biomolecular corona). Coenzyme corona formation is accompanied by the oxidation of NAD at biological concentrations through electron transfer. Similar phenomena are observed for NAD derivatives. Molecular dynamics simulations indicate that the adsorption of NAD onto CNTs is driven by interactions between the aromaphilic groups of NAD and the CNT surfaces, leading to coenzyme corona formation. Generally, in living biological systems, the balance of NAD redox (NADH/NAD+ redox) is maintained to sustain metabolism. The present results suggest that CNTs affect coenzyme-dependent metabolic reactions by disrupting the redox balance through coenzyme corona formation and subsequent coenzyme oxidation. The proposed molecular mechanism not only advances the fundamental understanding of the biological impact of CNTs in terms of metabolism but also contributes to biological CNT applications.


Assuntos
Coenzimas , Nanotubos de Carbono , Coenzimas/metabolismo , NAD/metabolismo , Oxirredução , Transporte de Elétrons
14.
J Orthop Sci ; 28(2): 358-363, 2023 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-34998617

RESUMO

BACKGROUND: The incidence of throwing-related elbow injuries is still rising. The study aimed to enhance the pathology of acute medial elbow injuries among young Little Leaguers by examining the medial elbows of symptomatic 9-10 years old Little Leaguers using High-Definition Magnetic Resonance Images (HDMRI), which uses a small-diameter surface coil on the target area, leading to greater image resolution. METHOD: We identified Little Leaguers aged 9-10 years old. To minimize the detection of the chronic adaptative changes, players who experienced the medial elbow pain previously and whose HDMRI had not been taken within 4 weeks from the onset of medial elbow pain were excluded. This study considered 21 players, and the mean age was 9.4 ± 0.5 years. RESULT: The fragmentation of the medial epicondyle apophysis via HDMRI was found in 15 elbows (71.4%), while the avulsion was seen in three cases. The signal hyperintensity at the medial epicondyle apophysis was observed in 2 cases. Our data showed abnormal changes to the medial epicondyle apophysis and surrounding structures, such as the ulnar collateral ligament (UCL), flexor-pronator tendons or the coronoid process of the ulna. We detected 11 abnormalities on X-ray imaging, while 20 subjects showed some abnormal findings via HDMRI. DISCUSSION: The current study showed that initial medial elbow injury in Little Leaguers without a history of previous elbow injury could be attributed to multi-structure injury. Over 90% of subjects were injured in the perichondrium, while 71.4% demonstrated a fragmentation of the secondary ossification center, and 14.3% experienced an avulsion of the medial epicondyle apophysis. Because the injuries were not limited to bony structures, HDMRI may be beneficial for the appropriate evaluation of medial elbow pain. The pathology of initial medial elbow injuries in young baseball players may be due to acute trauma instead of repetitive microtrauma.


Assuntos
Beisebol , Ligamentos Colaterais , Lesões no Cotovelo , Articulação do Cotovelo , Humanos , Criança , Cotovelo/diagnóstico por imagem , Articulação do Cotovelo/diagnóstico por imagem , Articulação do Cotovelo/patologia , Imageamento por Ressonância Magnética , Dor , Artralgia , Beisebol/lesões , Ligamentos Colaterais/lesões
15.
Food Chem Toxicol ; 170: 113508, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36336192

RESUMO

The increasing prevalence of mental disorders, such as depression, is currently a serious public health concern. Microalgae are a diverse group of organisms that contain many bioactive compounds such as polyunsaturated fatty acids and polyphenols. These compounds can exhibit many health benefits such as antioxidative, anti-inflammatory, anticancer, and anti-obesity effects. In the present study, we focused on microalgal (Botryococcus terribilis) extract (ME) rich in Me-meijicoccene (MM), a novel compound. Our results showed that pretreatment of SH-SY5Y cells with ME and MM ameliorated dexamethasone (depression-causing substance)-induced cytotoxicity. The results of the tail suspension test (TST) indicated that ME (50 mg/kg) induced antidepressant-like activity in TST-stressed mice. Our microarray analysis revealed that ME upregulated neurotransmitter-related gene (neurotransmitter secretion) expression and downregulated neuroinflammatory-related gene (chemokine-mediated signaling) expression in the cerebral cortex. ME also induced an increase in neurotransmitter and brain-derived neurotrophic factor levels, and a decrease in corticosterone and pro-inflammatory cytokine levels in the serum, cerebral cortex, and hypothalamus. Altogether, our study is the first to report that 50 mg/kg ME (not 100 mg/kg) exerts antidepressant-like effects via regulating neuroinflammation and modulating neurotransmitter systems in the mouse brain, highlighting the prospects of ME in the treatment of depressive disorders of a psychosocial nature.


Assuntos
Microalgas , Neuroblastoma , Animais , Humanos , Camundongos , Microalgas/metabolismo , Depressão/tratamento farmacológico , Doenças Neuroinflamatórias , Neuroblastoma/metabolismo , Antidepressivos/farmacologia , Fator Neurotrófico Derivado do Encéfalo/metabolismo , Extratos Vegetais/farmacologia , Extratos Vegetais/metabolismo , Neurotransmissores/metabolismo , Hipocampo , Modelos Animais de Doenças
16.
JBJS Case Connect ; 12(3)2022 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-36137070

RESUMO

CASE: We report a case of flexor pollicis longus (FPL) tendon rupture and carpal tunnel syndrome due to scaphoid nonunion advanced collapse deformity. Intraoperative findings showed disruption of the palmar joint capsule and a sharp proximal bone fragment protruding into the carpal tunnel. Removal of this proximal fragment and tendon grafting were performed. At the postoperative 2-year follow-up, the patient had no wrist pain, finger numbness, or restriction of thumb motion. CONCLUSION: Our results suggest that minimally invasive surgical procedures, such as proximal pole or osteophyte resection, might be optimal choices for early rehabilitation after tendon repair in cases of FPL tendon rupture due to asymptomatic scaphoid nonunion.


Assuntos
Síndrome do Túnel Carpal , Traumatismos do Antebraço , Traumatismos da Mão , Osso Escafoide , Traumatismos dos Tendões , Traumatismos do Punho , Síndrome do Túnel Carpal/cirurgia , Humanos , Osso Escafoide/diagnóstico por imagem , Osso Escafoide/cirurgia , Traumatismos dos Tendões/complicações , Traumatismos dos Tendões/cirurgia , Tendões , Traumatismos do Punho/complicações
17.
Sci Rep ; 12(1): 13718, 2022 08 12.
Artigo em Inglês | MEDLINE | ID: mdl-35962177

RESUMO

Since liquid-liquid phase separation (LLPS) of proteins is governed by their intrinsically disordered regions (IDRs), it can be controlled by LLPS-regulators that bind to the IDRs. The artificial design of LLPS-regulators based on this mechanism can be leveraged in biological and therapeutic applications. However, the fabrication of artificial LLPS-regulators remains challenging. Peptides are promising candidates for artificial LLPS-regulators because of their ability to potentially bind to IDRs complementarily. In this study, we provide a rational peptide design methodology for targeting IDRs based on residue-residue contact energy obtained using molecular dynamics (MD) simulations. This methodology provides rational peptide sequences that function as LLPS regulators. The peptides designed with the MD-based contact energy showed dissociation constants of 35-280 nM for the N-terminal IDR of the tumor suppressor p53, which are significantly lower than the dissociation constants of peptides designed with the conventional 3D structure-based energy, demonstrating the validity of the present peptide design methodology. Importantly, all of the designed peptides enhanced p53 droplet formation. The droplet-forming peptides were converted to droplet-deforming peptides by fusing maltose-binding protein (a soluble tag) to the designed peptides. Thus, the present peptide design methodology for targeting IDRs is useful for regulating droplet formation.


Assuntos
Proteínas Intrinsicamente Desordenadas , Proteínas Intrinsicamente Desordenadas/química , Simulação de Dinâmica Molecular , Peptídeos/metabolismo , Fenômenos Físicos , Proteína Supressora de Tumor p53/metabolismo
18.
J Chromatogr A ; 1676: 463277, 2022 Aug 02.
Artigo em Inglês | MEDLINE | ID: mdl-35809525

RESUMO

Immobilized metal ion affinity chromatography (IMAC) is useful in purification of histidine-tagged or histidine-rich proteins and peptides from a variety of hosts. However, phenolic compounds including polyphenols interfere with IMAC due to their high affinities for the transition metals immobilized on the column resins, which hampers the purification of proteins from plant-based host systems. In contrast to extensive knowledge of the mechanism of the interactions between phenolic compounds and transition metal ions in solution, an understanding of the interactions on the columns, where transition metal ions are immobilized on the resins, remains elusive. This study systematically investigated the affinity of phenolic compounds for transition metal ions by varying the number and position of phenolic hydroxyl groups (OH groups) and using different transition metals-Fe(II), Cu(II) and Ni(II)-on various IMACs, in which the columns were fabricated by equilibrating the cation-exchange column with transition metal solutions. It was found that the more OH groups the aromatic compounds have, the higher the affinity for transition metal ions; in particular, methyl gallate and pyrogallol were permanently bound to the IMAC column, which reflected coordinate bond formation with the transition metal ions. Importantly, the phenolic compounds showed no obvious affinity for the Ni(II)-IMAC column, in contrast to the Fe(II)- and Cu(II)-IMAC columns, whereas imidazole and histidine-tagged proteins showed evident binding to the Ni(II)-IMAC column. Ni(II)-IMAC should thus be especially effective in isolating histidine-tagged and histidine-rich species from phenolic compound-containing systems. These results indicate that the affinity between phenolic compounds and transition metal ions on the column is consistent with the results in solution. They also provide a comprehensive view for devising strategies to improve IMAC purification of target proteins and peptides from samples containing phenolic compounds.


Assuntos
Histidina , Peptídeos , Cátions , Cromatografia de Afinidade/métodos , Compostos Ferrosos , Histidina/química
19.
Artigo em Inglês | MEDLINE | ID: mdl-35822632

RESUMO

Nanoparticles are readily coated by proteins in biological systems. The protein layers on the nanoparticles, which are called the protein corona, influence the biological impacts of the nanoparticles, including internalization into cells and cytotoxicity. This study expands the scope of the nanoparticle's protein corona for exogenous artificial nanoparticles to that for exogenous proteinaceous nanoparticles. Specifically, this study addresses the formation of protein coronas on nanoscale human antibody aggregates with a radius of approximately 20-40 nm, where the antibody aggregates were induced by a pH shift from low to neutral pH. The size of the human immunoglobulin G (hIgG) aggregates grew to approximately 25 times the original size in the presence of human serum albumin (HSA). This size evolution was ascribed to the association of the hIgG aggregates, which was triggered by the formation of the hIgG aggregate's protein corona, i.e., protein's protein corona, consisting of the adsorbed HSA molecules. Because hIgG aggregate association was significantly reduced by the addition of 30-150 mM NaCl, it was attributed to electrostatic attraction, which was supported by molecular dynamics (MD) simulations. Currently, the use of antibodies as biopharmaceuticals is concerning because of undesired immune responses caused by antibody aggregates that are typically generated by a pH shift during the antibody purification process. The present findings suggest that nanoscale antibody aggregates form protein coronas induced by HSA and the resulting nanoscale antibody-HSA complexes are stable in blood containing approximately 150 mM salt ions, at least in terms of the size evolution. Mechanistic insights into protein corona formation on nanoscale antibody aggregates are useful for understanding the unintentional biological impacts of antibody drugs.

20.
Biomedicines ; 10(5)2022 May 20.
Artigo em Inglês | MEDLINE | ID: mdl-35625924

RESUMO

A person's quality of life can be adversely affected by hair loss. Microalgae are widely recognized for their abundance and rich functional components. Here, we evaluated the hair growth effect of a green alga, Botryococcus terribilis (B. terribilis), in vitro using hair follicle dermal papilla cells (HFDPCs). We isolated two types of cells from B. terribilis-green and orange cells, obtained from two different culture conditions. Microarray and real time-PCR results revealed that both cell types stimulated the expression of several pathways and genes associated with different aspect of the hair follicle cycle. Additionally, we demonstrated B. terribilis' effect on collagen and keratin synthesis and inflammation reduction. We successfully isolated a novel compound, methylated-meijicoccene (me-meijicoccene), and C32 botryococcene from B. terribilis to validate their promising effects. Our study revealed that treatment with the two compounds had no cytotoxic effect on HFDPCs and significantly enhanced the gene expression levels of hair growth markers at low concentrations. Our study provides the first evidence of the underlying hair growth promoting effect of B. terribilis and its novel compound, me-meijicoccene, and C32 botryococcene.

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