35 research outputs found
DRUG DISCOVERY FROM PLANTS: TRADITIONAL HERBAL REMEDIES AS SOURCE FOR NEW LEAD COMPOUNDS
Resposta de anticorpos IgE, IgG1 e IgG4 aos componentes ligantes de Concanavalina A isolados de Blomia tropicalis (Acari: Echimyopodidae) em indivĂduos alĂ©rgicos e nĂŁo alĂ©rgicos
Blomia tropicalis (Bt) and Dermatophagoides pteronyssinus (Dp) are the most
prevalent house dust mites in tropical countries and associated with allergic
diseases. Glycosilated antigens are highly immunogenic and involved in different
pathologies, including allergies. The aims of this study were to evaluate IgE, IgG1,
and IgG4 responses to Concanavalin A-binding components (Bt-ConA) isolated from
Bt extract in sera of allergic and non-allergic subjects and to analyze the crossreactivity
with Dp extract. Also, IgE-, IgG1- and IgG4-reactive antigenic components
from both extracts were analyzed by Immunoblotting. Bt-ConA was obtained from Btwhole
extract fractionated on Con-A-Sepharose affinity chromatography and both
extracts were evaluated in SDS-PAGE and ELISA for IgE, IgG1, and IgG4 in sera of
121 patients with allergic rhinitis and 36 non-allergic subjects. Subjects were skin
prick tested (SPT) to Bt-whole extract. Inhibition and immunoblotting test were
performed to analyze IgE, IgG1, and IgG4 responses to both extracts. SPT showed
that 58% of patients were sensitized to Bt (Bt+), with 52% reactive to both mites (Bt
and Dp) and 6% to Bt only. A broad spectrum of proteins (14-152kDa) were
visualized in Bt-whole extract and components >27kDa in Bt-ConA extract. ELISA
showed a similar profile of IgE, IgG1 and IgG4 levels to Bt-whole and Bt-ConA
extracts in different groups, although Bt+ patients showed a lower IgG4 reactivity to
Bt-ConA extract. Specific IgG1 levels were higher in Bt+ patients than control
subjects, and IgG4 levels showed no significant difference among the groups. ELISA
inhibition showed a partial IgE and total IgG1 and IgG4 cross-reactivity with Dp
extract for Bt-whole and Bt-ConA extracts. Immunoblotting revealed ten antigenic
components in Bt-whole extract (14-152kDa) recognized by IgE and IgG4 antibodies
and five of these components (>50kDa) were recognized by IgG1 in Bt+ patients. Bt-
ConA extract showed eight antigenic components (27-152kDa), from which the 152
and 123kDa bands were predominantly recognized by IgE, and only three of these
components (93, 123 and 152kDa) were recognized by IgG1 while IgG4 antibodies
weakly recognized the 66 and 152kDa components in Bt+ patients. It can be
concluded that Con A-binding components isolated from Bt constitute major allergens
and are involved in both allergen sensitization (IgE response) and homeostase
maintenance (IgG1 and IgG4 responses) and that glycosilated components,
particularly those of high molecular weight, are preferentially recognized by IgE and
IgG1, but not by IgG4 antibodies.Mestre em CiĂȘncias da SaĂșdeBlomia tropicalis (Bt) e Dermatophagoides pteronyssinus (Dp) sĂŁo os
principais ĂĄcaros da poeira domiciliar em paĂses tropicais e subtropicais que estĂŁo
associados com doenças alĂ©rgicas. AntĂgenos glicosilados sĂŁo altamente
imunogĂȘnicos e estĂŁo envolvidos em diferentes patologias, incluindo alergias. Os
objetivos deste estudo foram avaliar a resposta de anticorpos IgE, IgG1 e IgG4 a
componentes ligantes de Concanavalina A (Bt-ConA) isolados do extrato Bt total em
soros de indivĂduos alĂ©rgicos e nĂŁo alĂ©rgicos, bem como a reatividade cruzada com
Dp e identificar por immunoblotting componentes protéicos de ambos extratos
reconhecidos por anticorpos IgE, IgG1 e IgG4. O extrato Bt-ConA foi obtido por
fracionamento do extrato Bt total em cromatografia de afinidade Con A-Sepharose e
ambos extratos foram avaliados em SDS-PAGE e ELISA para IgE, IgG1 e IgG4 em
121 soros de pacientes com rinite alĂ©rgica e 36 indivĂduos nĂŁo alĂ©rgicos. Testes
cutĂąneos de puntura (TCP) foram realizados em todos os indivĂduos do estudo.
Ensaios de inibição foram realizados para detecção de IgE, IgG1 e IgG4 especĂfica a
Bt total e Bt-ConA. Ensaios de immunoblotting foram realizados para detecção de
componentes antigĂȘnicos de ambos extratos reconhecidos por IgE, IgG1 e IgG4. Do
total de 121 pacientes, 58% mostraram-se sensibilizados a Bt (Bt+), com 52%
apresentando reatividade cutĂąnea a ambos ĂĄcaros (Bt e Dp) e apenas 6% somente
a Bt. Um largo espectro de proteĂnas (14-152kDa) foram visualizadas no extrato Bt
total e componentes acima de 27kDa na fração Bt-ConA. ELISA apresentou um perfil
similar nos nĂveis de IgE, IgG1 e IgG4 para os extratos Bt total e Bt-ConA em
diferentes grupos, embora pacientes do grupo Bt+ demonstraram uma reatividade
menor de IgG4 para o extrato Bt-ConA. NĂveis de IgG1 especĂfica foram maiores nos
pacientes Bt+ do que nos indivĂduos controles e os nĂveis de IgG4 nĂŁo apresentaram
diferença significativa entre os grupos. ELISA de inibição demonstrou uma
reatividade cruzada parcial com o extrato de Dp para IgE especĂfica e total para IgG1
e IgG4 especĂficas aos extratos de Bt total e Bt-ConA. Immunoblotting revelou dez
componentes antigĂȘnicos do extrato Bt total (14-152kDa) reconhecidos por
anticorpos IgE e IgG4 e cinco destes componentes (>50kDa) foram reconhecidos
por IgG1 em pacientes Bt+. O extrato Bt-ConA revelou oito componentes antigĂȘnicos
(27-152kDa), dos quais as bandas de 152 e 123kDa foram predominantemente
reconhecidos por IgE e apenas trĂȘs destes componentes (93, 123 e 152kDa) foram
reconhecidos por IgG1 enquanto anticorpos IgG4 reconheceram fracamente os
componentes de 66 e 152kDa em pacientes Bt+. Pode-se concluir que componentes
ligantes de Con-A isolados de Bt constituem alérgenos principais e estão envolvidos
tanto na sensibilização alergĂȘnica (resposta IgE) quanto na manutenção da
homeostase (respostas IgG1 e IgG4) e que componentes glicosilados,
particularmente os de alto peso molecular, sĂŁo preferencialmente reconhecidos por
anticorpos IgE e IgG1, mas nĂŁo por anticorpos IgG4
Quick development of an analytical enantioselective high performance liquid chromatography separation and preparative scale-up for the flavonoid Naringenin
Nickel quantification in serum by a validated sector-field inductively coupled plasma mass spectrometry method: Assessment of tentative reference values for an Italian population
An eco-friendly enantioselective access to (R)-Naringenin as inhibitor of proinflammatory cytokine release.
(R/S)-Naringenin is a flavanone, well-known for its beneficial health-related properties, such as anti-inflammatory activity. The preparative enantioselective chromatographic resolution of commercial (R/S)-Naringenin was performed on ChiralpakÂź AD-H column (500 x 50 mm I.D., dp = 20 ÎŒm) using methanol as eluent. The developed method is in accordance with the principles of green chemistry, since the environmental impact was lowered through the recycle of eluent. In this way, g-amounts of each enantiomer were recovered with high purity (chemical purity > 99%, enantiomeric excess > 94%). The racemic and enantiomeric Naringenin were subjected to an exhaustive in vitro investigation of anti-inflammatory activity, aimed at evaluating the relevance of chirality. The assay on cultured human peripheral blood mononuclear cells activated by Phytohemagglutinin A revealed that (R)-Naringenin is the most effective in inhibiting the T-cell proliferation, showed to be still active at lowest concentration. Moreover, (R)-Naringenin significantly reduced pro-inflammatory cytokines levels such as TNF-α and, with less potency, IL-6. These results evidenced the anti-inflammatory potential of Naringenin and the highest capacity of (R)-Naringenin to inhibit both in vitro PBMC proliferation and cytokines secretion at non toxic doses. Thus, (R)-Naringenin is a promising candidate for in vivo investigation
An eco-friendly enantioselective access to (R)-Naringenin as inhibitor of proinflammatory cytokine release.
(R/S)-Naringenin is a flavanone, well-known for its beneficial health-related properties, such as anti-inflammatory activity. The preparative enantioselective chromatographic resolution of commercial (R/S)-Naringenin was performed on ChiralpakÂź AD-H column (500 x 50 mm I.D., dp = 20 ÎŒm) using methanol as eluent. The developed method is in accordance with the principles of green chemistry, since the environmental impact was lowered through the recycle of eluent. In this way, g-amounts of each enantiomer were recovered with high purity (chemical purity > 99%, enantiomeric excess > 94%). The racemic and enantiomeric Naringenin were subjected to an exhaustive in vitro investigation of anti-inflammatory activity, aimed at evaluating the relevance of chirality. The assay on cultured human peripheral blood mononuclear cells activated by Phytohemagglutinin A revealed that (R)-Naringenin is the most effective in inhibiting the T-cell proliferation, showed to be still active at lowest concentration. Moreover, (R)-Naringenin significantly reduced pro-inflammatory cytokines levels such as TNF-α and, with less potency, IL-6. These results evidenced the anti-inflammatory potential of Naringenin and the highest capacity of (R)-Naringenin to inhibit both in vitro PBMC proliferation and cytokines secretion at non toxic doses. Thus, (R)-Naringenin is a promising candidate for in vivo investigation
Phytochemical and biological profile of Eremurus persicus Boiss root extract: isolation of the main phytocomponents.
Identification of peptides with ELAV-like mRNA-stabilizing effect: an integrated in vitro/in silico approach
Embryonic lethal abnormal vision (ELAV) proteins are RNA-binding proteins that bind specific adenine and uridine-rich elements mainly located in the 3'-untranslated region of target mRNAs, preventing their otherwise rapid degradation and thus increasing gene expression. Starting from our previous discovery and applying an integrated in vitro/in silico approach, herein we report a deeper understanding of the mRNA-stabilizing activity of four peptides derived from the ELAV proteins structure. The stabilizing effect on the VEGF transcript (mRNAVEGF ) exerted by each peptide, tested individually, was initially evaluated, and no effects were evidenced. Hence, the biological effects of all peptides couples were investigated. Interestingly, in accordance with preliminary molecular dynamics results, only one of all possible peptide couples resulted highly effective in stabilizing mRNAVEGF . These two peptides were thus identified as valuable starting point for designing small molecules with ELAV-mimicking properties
Are sigma modulators an effective opportunity for cancer treatment? A patent overview (1996-2016).
Introduction: Although several molecular targets against cancer have been identified, there is a
continuous need for new therapeutic strategies. Sigma Receptors (SRs) overexpression has been
recently associated with different cancer conditions. Therefore, novel anticancer agents targeting SRs
may increase the specificity of therapies, overcoming some of the common drawbacks of conventional
chemotherapy.
Areas covered: The present review focuses on patent documents disclosing SR modulators with
possible application in cancer therapy and diagnosis. The analysis reviews patents of the last two
decades (1996â2016); patents were grouped according to target subtypes (S1R, S2R, pan-SRs) and
relevant Applicants. The literature was searched through Espacenet, ISI Web, PatentScope and PubMed
databases.
Expert opinion: The number of patents related to SRs and cancer has increased in the last twenty
years, confirming the importance of this receptor family as valuable target against neoplasias. Despite
their short history in the cancer scenario, many SR modulators are at pre-clinical stage and one is
undergoing a phase II clinical trial. SRs ligands may represent a powerful source of innovative antitumor
therapeutics. Further investigation is needed for validating SR modulators as anti-cancer drugs. We
strongly hope that this review could stimulate the interest of both Academia and pharmaceutical
companies