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1.
Protein Expr Purif ; 166: 105505, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31563543

RESUMO

Recombinant antibodies can be expressed as fusion constructs in combination with tags which simplify their engineering into reliable and homogeneous immunoreagents by allowing site-specific, 1:1 functionalization. Several tags and corresponding reagents for recombinant protein derivatization have been proposed but benchmarking surveys for the evaluation of their effect on the characteristics of recombinant antibodies have not been reported. In this work we evaluated the impact on expression yields, shelf-stability, thermostability and binding affinity of a set of C-terminal tags fused to the same anti-Her2 nanobody. Furthermore, we assessed the efficiency of the derivatization process. The constructs always bore a 6xHis tag plus either the controls (EGFP and C-tag) or CLIP, HALO, AviTag, the LEPTG sequence recognized by Sortase A (Sortase tag), or a free cysteine. The advantages and drawbacks of the different systems were analyzed and discussed.


Assuntos
Proteínas Recombinantes de Fusão/genética , Anticorpos de Domínio Único/genética , Ligação Competitiva , Cisteína/metabolismo , Escherichia coli , Proteínas de Escherichia coli/química , Proteínas de Escherichia coli/genética , Vetores Genéticos/genética , Proteínas de Fluorescência Verde/química , Proteínas de Fluorescência Verde/genética , Oxirredutases/química , Oxirredutases/genética , Isomerases de Dissulfetos de Proteínas/química , Isomerases de Dissulfetos de Proteínas/genética , Estabilidade Proteica , Receptor ErbB-2/química , Receptor ErbB-2/genética , Proteínas Recombinantes de Fusão/química , Anticorpos de Domínio Único/química
2.
Org Biomol Chem ; 14(21): 4853-8, 2016 Jun 07.
Artigo em Inglês | MEDLINE | ID: mdl-27151329

RESUMO

JNJ-26990990 ((benzo[b]thien-3-yl)methyl)sulfamide, a sulfamide derivative structurally related to the antiepileptic drug zonisamide, was reported to be devoid of carbonic anhydrase (CA, EC 4.2.1.1) inhibitory properties. Here we report that JNJ-26990990 and its S,S-dioxide analog significantly inhibit six human (h) isoforms, hCA I, II, VII, IX, XII and XIV, involved in crucial physiological processes. Inhibition and X-ray crystallographic data for the binding of the two compounds to these enzymes show significant similarity with the zonisamide inhibitory pattern. These findings prompted us to reconsider the structural/pharmacological requirements for designing effective antiepileptics possessing zinc-binding groups of the sulfamide, sulfamate or sulfonamide type in their molecules.


Assuntos
Anidrases Carbônicas/metabolismo , Óxidos/química , Sulfonamidas/química , Sulfonamidas/farmacologia , Enxofre/química , Tiofenos/química , Tiofenos/farmacologia , Anticonvulsivantes/química , Anticonvulsivantes/farmacologia , Inibidores da Anidrase Carbônica/química , Inibidores da Anidrase Carbônica/farmacologia , Anidrases Carbônicas/química , Cristalografia por Raios X , Humanos , Modelos Moleculares , Conformação Proteica , Soluções
3.
Foot Ankle Int ; 32(5): S526-32, 2011 May.
Artigo em Inglês | MEDLINE | ID: mdl-21733462

RESUMO

BACKGROUND: Numerous studies document the high incidence of this type of fracture of the foot in childhood due to the presence of the growth plate of the apophysis of the fifth metatarsal. Our aim in this study was to evaluate the presence and morphology of the growth nucleus and correlate its presence with fractures of the apophysis of the fifth metatarsal. MATERIALS AND METHODS: Between 2001 and 2003, we reviewed the radiographs of 481 patients (558 feet) between 6 months and 16 years that were diagnosed with a fracture. We evaluated for the presence and morphology of the growth nucleus of the base of the fifth metatarsal in the 7- to 16-year age group because this is the time interval in which the nucleus becomes visible radiographically. RESULTS: We identified the nucleus of the base of the fifth metatarsal in 115 patients for a total of 132 feet. A fracture of the fifth metatarsal was found in 12.8%. A misdiagnosis had been made in 47%. Misdiagnosis was strictly related to the presence of the growth nucleus of the apophysis except two cases where an accessory bone was present. CONCLUSIONS: Our study demonstrated a much higher incidence of the presence of the growth nucleus than documented in previous reports and that its morphology varied widely in relation to the stage of maturity of the patient and the radiographic technique. Its presence led to a high rate of misdiagnosis of a fracture.


Assuntos
Traumatismos do Pé/diagnóstico por imagem , Fraturas Ósseas/diagnóstico por imagem , Lâmina de Crescimento/diagnóstico por imagem , Ossos do Metatarso/lesões , Adolescente , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Masculino , Ossos do Metatarso/diagnóstico por imagem , Radiografia , Falanges dos Dedos do Pé/lesões
4.
In Vivo ; 26(2): 299-304, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22351673

RESUMO

Human mesenchymal stem cells (hMSC) have become an important resource in developing strategies for regenerative medicine and tissue engineering, owing to their ability to renew and their potential for differentiation into cells of various types of tissues. Pulsed electromagnetic field (PEMF) stimulation has been used for several years in the treatment of fracture healing, with clinical beneficial effects, and several studies have demonstrated its capacity to increase bone tissue regeneration. In the present study, stromal cells of human bone marrow (BMSC), obtained from healthy donors, were appropriately expanded and underwent PEMF stimulation eight hours a day for fourteen days. Parameters such as proliferation and differentiation ability were evaluated on stimulated cultures. The evaluation of the marker expression was performed by RT-PCR for osteocalcin, by alkaline phosphatase quantitation and by histochemical stains. The results we obtained showed that BMSC treated with PEMF begin differentiation earlier than untreated BMSC, as shown by the markers used. The data show that PEMF is able to increase the osteogenic differentiation potential in adult mesenchymal cells isolated from young patients.


Assuntos
Campos Eletromagnéticos , Células-Tronco Mesenquimais/efeitos da radiação , Osteócitos/citologia , Osteogênese/efeitos da radiação , Adulto , Fosfatase Alcalina/biossíntese , Fosfatase Alcalina/genética , Células da Medula Óssea/citologia , Células da Medula Óssea/efeitos da radiação , Calcificação Fisiológica/efeitos da radiação , Ciclo Celular/efeitos da radiação , Células Cultivadas/citologia , Células Cultivadas/efeitos da radiação , Meios de Cultura/farmacologia , Indução Enzimática/efeitos da radiação , Regulação da Expressão Gênica no Desenvolvimento/efeitos da radiação , Humanos , Células-Tronco Mesenquimais/citologia , Osteocalcina/biossíntese , Osteocalcina/genética , Osteogênese/efeitos dos fármacos , Células Estromais/citologia , Células Estromais/efeitos da radiação
5.
Infect Agent Cancer ; 4: 2, 2009 Jan 16.
Artigo em Inglês | MEDLINE | ID: mdl-19149879

RESUMO

Acute viral hepatitis B is cleared in more than 95% of patients, while the remainder ones may develop either chronic HBV infection or, rarely, fulminant hepatitis.Therefore there are elderly patients with severe acute HBV hepatitis caractherized by high serum bilirubin levels >15 mmole/dl, international normalized ratio (INR) with value more than 1.6; these patients are caractherized by a severe outcome of HBV infection.As known, outcome of infection and the pathogenesis of liver diseases are determined by viral and host factors, such as T reg lymphocytes.T regs may be associated with a negative immune response such as an inhibition of gamma- IFN secretion.The impact of viral load on antiviral T cell responses may play a critical role in these patients, influencing disease persistence and immune response.Antiviral drugs could be useful in these patients determing a possible down -regulation of T regs.

6.
Infect Agent Cancer ; 4: 5, 2009 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-19338663

RESUMO

Correction to Morelli G, Perrella A, Sbreglia C, Bellopede P, Riccio V, Perrella O: Antiviral therapy in acute viral hepatitis B: why and when. Infectious Agents and Cancer 2009, 4:2.

7.
Am J Med Genet A ; 124A(4): 407-10, 2004 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-14735591

RESUMO

We describe a family with a distinctive malformation of the hand consisting of the fusion of the 4th and the 5th metacarpal bones. Usually this anomaly is clinically recognizable by an ulnar deviation of the 5th finger; moreover, the 5th metacarpal is usually hypoplastic and the 5th ray is consequently short. There is, however, great variability in expression, so the degree of fusion may range from minimal to complete and also the external aspect of the hand may vary. This anomaly can be either isolated or part of a syndrome. For the isolated form, two possible hereditary mechanisms have been proposed: autosomal dominant and X-linked recessive. Our family is consistent with the latter, with only affected males and no instances of male-to-male transmission. Of note, there are very few X-linked recessive disorders that affect the hand in a such a specific way.


Assuntos
Doenças Genéticas Ligadas ao Cromossomo X/diagnóstico , Deformidades Congênitas da Mão/diagnóstico , Metacarpo/anormalidades , Adolescente , Cromossomos Humanos X , Genes Recessivos , Deformidades Congênitas da Mão/genética , Deformidades Congênitas da Mão/cirurgia , Humanos , Masculino , Linhagem
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