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1.
J Trace Elem Med Biol ; 84: 127457, 2024 Apr 27.
Artigo em Inglês | MEDLINE | ID: mdl-38692231

RESUMO

The aim of this paper was the quantitative determination of some macro and trace elements, especially potentially toxic elements in the samples of infant baby formulae and baby food cereals commercially available in Serbia using the inductively coupled plasma optical emission spectrometry (ICP OES) method. Among the macro elements, K is the most abundant in all infant formulae samples, followed by Ca, P, Na and Mg. On the other hand, the analysis of food cereals showed that P is presents in the highest contents, followed by K, Ca, Na, and Mg. Potentially toxic elements As, Pb, Hg, and Cd were not detected in any sample of infant formulae, while Cd was detected and quantified in cereal foods. Also, the calculated values of Estimated Tolerable Weekly Intake (ETWI) as well as the Estimated Tolerable Monthly Intake (ETMI) were lower than recommended values for a tolerable weekly intake (TWI) and provisional tolerable monthly intake (PTMI).

2.
J Clin Med ; 12(4)2023 Feb 17.
Artigo em Inglês | MEDLINE | ID: mdl-36836134

RESUMO

As the key enzyme mediating ribonucleotide excision repair, RNase H2 is essential for the removal of single ribonucleotides from DNA in order to prevent genome damage. Loss of RNase H2 activity directly contributes to the pathogenesis of autoinflammatory and autoimmune diseases and might further play a role in ageing and neurodegeneration. Moreover, RNase H2 activity is a potential diagnostic and prognostic marker in several types of cancer. Until today, no method for quantification of RNase H2 activity has been validated for the clinical setting. Herein, validation and benchmarks of a FRET-based whole-cell lysate RNase H2 activity assay are presented, including standard conditions and procedures to calculate standardized RNase H2 activity. Spanning a wide working range, the assay is applicable to various human cell or tissue samples with overall methodological assay variability from 8.6% to 16%. Using our assay, we found RNase H2 activity was reduced in lymphocytes of two patients with systemic lupus erythematosus and one with systemic sclerosis carrying heterozygous mutations in one of the RNASEH2 genes. Implementation of larger control groups will help to assess the diagnostic and prognostic value of clinical screening for RNase H2 activity in the future.

3.
Sci Immunol ; 7(70): eabl7482, 2022 04 15.
Artigo em Inglês | MEDLINE | ID: mdl-35427180

RESUMO

Macrophages populate every organ during homeostasis and disease, displaying features of tissue imprinting and heterogeneous activation. The disconnected picture of macrophage biology that has emerged from these observations is a barrier for integration across models or with in vitro macrophage activation paradigms. We set out to contextualize macrophage heterogeneity across mouse tissues and inflammatory conditions, specifically aiming to define a common framework of macrophage activation. We built a predictive model with which we mapped the activation of macrophages across 12 tissues and 25 biological conditions, finding a notable commonality and finite number of transcriptional profiles, in particular among infiltrating macrophages, which we modeled as defined stages along four conserved activation paths. These activation paths include a "phagocytic" regulatory path, an "inflammatory" cytokine-producing path, an "oxidative stress" antimicrobial path, or a "remodeling" extracellular matrix deposition path. We verified this model with adoptive cell transfer experiments and identified transient RELMɑ expression as a feature of monocyte-derived macrophage tissue engraftment. We propose that this integrative approach of macrophage classification allows the establishment of a common predictive framework of monocyte-derived macrophage activation in inflammation and homeostasis.


Assuntos
Ativação de Macrófagos , Macrófagos , Animais , Citocinas/metabolismo , Homeostase , Inflamação/metabolismo , Camundongos
4.
Food Chem ; 378: 132113, 2022 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-35033722

RESUMO

The study aimed to analyze trace elements content in baby purees and fruit juices and to evaluate the health risk of young children. The average daily dose, hazard quotient, hazard index and total diet hazard quotient were calculated to assess the potential health risk on per capita and consumers only groups of infants and toddlers. There was no significant health risk for studied groups regarding the intake of trace elements via purees and juices consumption. Health risk for lead was not estimated since the oral reference dose for this metal was not yet established and PTWI value was withdrawn. The average daily dose of lead for infants (0.32 - 0.46 µg/kg bw/day) and toddlers (2.01 - 2.29 µg/kg bw/day) were in accordance with the daily lead exposure intervals estimated by EFSA. Applying statistical analysis, the products were classified into three groups according to the content of trace elements.


Assuntos
Oligoelementos , Pré-Escolar , Dieta , Sucos de Frutas e Vegetais , Humanos , Lactente , Medição de Risco
5.
EBioMedicine ; 66: 103314, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33813142

RESUMO

BACKGROUND: Nucleic acids are potent stimulators of type I interferon (IFN-I) and antiviral defense, but may also promote pathological inflammation. A range of diseases are characterized by elevated IFN-I, including systemic lupus erythematosus (lupus). The DNA-activated cGAS-STING pathway is a major IFN-I-inducing pathway, and activation of signaling is dependent on trafficking of STING from the ER to the Golgi. METHODS: Here we used cell culture systems, a mouse lupus model, and material from lupus patients, to explore the mode of action of a STING antagonistic peptide, and its ability to modulate disease processes. FINDINGS: We report that the peptide ISD017 selectively inhibits all known down-stream activities of STING, including IFN-I, inflammatory cytokines, autophagy, and apoptosis. ISD017 blocks the essential trafficking of STING from the ER to Golgi through a mechanism dependent on the STING ER retention factor STIM1. Importantly, ISD017 blocks STING activity in vivo and ameliorates disease development in a mouse model for lupus. Finally, ISD017 treatment blocks pathological cytokine responses in cells from lupus patients with elevated IFN-I levels. INTERPRETATION: These data hold promise for beneficial use of STING-targeting therapy in lupus. FUNDING: The Novo Nordisk Foundation, The European Research Council, The Lundbeck Foundation, European Union under the Horizon 2020 Research, Deutsche Forschungsgemeinschaft, Chulalongkorn University.


Assuntos
Retículo Endoplasmático/metabolismo , Complexo de Golgi/metabolismo , Lúpus Eritematoso Sistêmico/etiologia , Lúpus Eritematoso Sistêmico/metabolismo , Proteínas de Membrana/antagonistas & inibidores , Proteínas de Neoplasias/metabolismo , Molécula 1 de Interação Estromal/metabolismo , Animais , Linhagem Celular , Modelos Animais de Doenças , Suscetibilidade a Doenças , Vesículas Extracelulares/metabolismo , Expressão Gênica , Humanos , Lúpus Eritematoso Sistêmico/tratamento farmacológico , Lúpus Eritematoso Sistêmico/patologia , Camundongos , Camundongos Knockout , Transporte Proteico/efeitos dos fármacos
6.
Vaccines (Basel) ; 8(4)2020 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-33271962

RESUMO

Vaccines can have heterologous effects on the immune system, i.e., effects other than triggering an immune response against the disease targeted by the vaccine. We investigated whether monoclonal antibodies (mAbs) specific for tetanus could cross-react with Chlamydia and confer heterologous protection against chlamydial infection. The capability of two tetanus-specific mAbs, namely mAb26 and mAb51, to prevent chlamydial infection has been assessed: (i) in vitro, by performing a neutralization assay using human conjunctival epithelial (HCjE) cells infected with Chlamydia trachomatis serovar B, and (ii) in vivo, by using a guinea pig model of Chlamydiacaviae-induced inclusion conjunctivitis. The mAb26 has been superior in comparison with mAb51 in the prevention of chlamydial infection in HCjE cells. The mAb26 has conferred ≈40% inhibition of the infection, compared to less than 5% inhibition in the presence of the mAb51. In vivo, mAb26 significantly diminished ocular pathology intensity in guinea pigs infected with C. caviae compared to either the mAb51-treated or sham-treated guinea pigs. Our data provide insights that tetanus immunization generates antibodies which induce heterologous chlamydial immunity and promote protection beyond the intended target pathogen.

7.
Saudi Pharm J ; 28(3): 246-254, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-32194325

RESUMO

In this study, a series of synthesized 3-(4-substituted benzyl)-5-isopropyl-5-phenylhydantoin derivatives as a potential antiproliferative and antimigratory agents were investigated. The possible antitumor mechanisms of investigated hydantoin derivatives were examined on human breast cancer cell line MDA-MB-231. The cells were treated with different concentrations of compounds (from 0.01 µM to 100 µM) during 24 h and 72 h. The proliferation index, nitric oxide production, apoptosis rate, and migration capacity were measured. The cell invasion potential was examined by measuring the level of MMP-9 and COX-2 gene expression. All tested compounds expressed antiproliferative activity and induced dose- and time-dependent increase in the level of nitrites. The investigated molecules significantly decreased cell survival rate, migration capacity and the expression levels of genes included in the process of tumor invasion. Obtained data suggest that the tested hydantoin derivatives express considerable antitumor activity by reducing cell division rate, elevating apoptosis level, and inhibiting the motility and invasiveness of breast cancer cells. The results obtained in this study indicate that investigated compounds express potential as a novel chemotherapeutic agents against breast cancer growth and progression.

8.
J Sci Food Agric ; 99(12): 5424-5431, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31077389

RESUMO

BACKGROUND: Pesticides are potentially toxic to humans and can produce both acute and chronic health effects, depending on the quantity and the ways in which a person is exposed. Exposure to pesticides can cause serious health problems. Infants and young children are particularly sensitive to these contaminants because their brains and organ systems are not fully developed. For this reason, it is important to determine the quantities of pesticides in baby food. RESULTS: The aim of this study was to develop a kinetic-spectrophotometric method for atrazine determination and to apply it to determine pesticide in baby-food samples, using solid-phase extraction (SPE) followed by the kinetic-spectrophotometric method and the high-performance liquid chromatography (HPLC) method. This method is based on the inhibition effect of atrazine (the oxidation of sulfanilic acid (SA) by hydrogen peroxide in the alkaline medium in the presence of the Co2+ ion). Under the experimental conditions used, atrazine showed a linear dynamic range of 0.5 to 5.0 µg mL-1 , and from 5.0 to 70.00 µg mL-1 with relative standard deviations (RSD) from 1.91% to 9.41%. The limit of detection and the limit of quantification were 0.074 and 0.225 µg mL-1 , respectively. The kinetic method was successfully applied to determine the atrazine concentration in spiked samples after SPE of samples. High-performance liquid chromatography was used to verify the results. CONCLUSION: The proposed method is highly sensitive, simple, easy, requires cheap reagents, and leads to good recovery levels. It is linear, precise, and accurate. It can be used successfully for the routine analysis of atrazine in infant formulae and cereal-based food samples. © 2019 Society of Chemical Industry.


Assuntos
Atrazina/química , Grão Comestível/química , Contaminação de Alimentos/análise , Fórmulas Infantis/química , Resíduos de Praguicidas/química , Espectrofotometria/métodos , Cromatografia Líquida de Alta Pressão
9.
Acta Chim Slov ; 66(4): 802-813, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34057496

RESUMO

This study aimed to optimize and validate the inductively coupled plasma optical emission spectrometric method (ICP OES) for the simultaneous determination of eleven potentially toxic elements (Al, Cd, Cr, Co, Cu, Ni, Pb, Fe, Sb, Mn, and Zn) in lipstick samples. The method was evaluated by applying the standard addition method. The recoveries for all elements in lipsticks were between 90% and 110%, except for Cd and Pb they were <90% and >110%, respectively. The health risk assessment was determined by calculating the average daily intake (ADD), hazard quotient (HQ), and hazard index (HI). The highest mean value for ADD was for Fe (4.8×10-1 mg kg-1 day-1), and the lowest was for Co (9.3×10-6 mg kg-1 day-1). There was no significant toxic health risk for any of the elements (HQ < 1), except for Fe (HQ < 3) which indicates a potential health risk. Based on PCA, all potentially toxic elements have been classified in the three groups. The first group includes Fe, the second includes Al, and all other elements belong to the third group. The cluster analysis of the elements provided the identical grouping that was obtained on the basis of PCA. Two separate clusters were obtained when cluster analysis was applied to the analyzed samples. The first cluster contained the only sample that was brown. The second cluster was divided into two sub-clusters. The first sub-cluster included the samples belonging to category I regarding the price, while the second sub-cluster included the samples belonging to category II and III regarding the price.

10.
J Appl Phycol ; 30(4): 2601-2610, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30147240

RESUMO

Ocular chlamydial infections with the ocular serovars A, B, Ba, and C of Chlamydia trachomatis represent the world's leading cause of infectious blindness. Carrageenans are naturally occurring, sulfated polysaccharides generally considered safe for food and topical applications. Carrageenans can inhibit infection caused by a variety of viruses and bacteria. To investigate whether iota-carrageenan (I-C) isolated from the red alga Chondrus crispus could prevent ocular chlamydial infection, we assessed if targeted treatment of the conjunctival mucosa with I-C affects chlamydial attachment, entry, and replication in the host cell. Immortalized human conjunctival epithelial cells were treated with I-C prior to C. trachomatis infection and analyzed by flow cytometry and immunofluorescence microscopy. In vivo effects were evaluated in an ocular guinea pig inclusion conjunctivitis model. Ocular pathology was graded daily, and chlamydial clearance was investigated. Our study showed that I-C reduces the infectivity of C. trachomatis in vitro. In vivo results showed a slight reduced ocular pathology and significantly less shedding of infectious elementary bodies by infected animals. Our results indicate that I-C could be a promising agent to reduce the transmission of ocular chlamydial infection and opens perspectives to develop prophylactic approaches to block C. trachomatis entry into the host cell.

11.
Vaccine ; 36(26): 3764-3771, 2018 06 18.
Artigo em Inglês | MEDLINE | ID: mdl-29773320

RESUMO

Oligoclonal combinations of several monoclonal antibodies (MAbs) are being considered for the treatment of various infectious pathologies. These combinations are less sensitive to antigen structural changes than individual MAbs; at the same time, their characteristics can be more efficiently controlled than those of polyclonal antibodies. The main goal of this study was to evaluate the binding characteristics of six biclonal equimolar preparations (BEP) of tetanus toxin (TeNT)-specific MAbs and to investigate how the MAb combination influences the BEPs' protective capacity. We show that a combination of TeNT-specific MAbs, which not only bind TeNT but also exert positive cooperative effects, results in a BEP with superior binding characteristics and protective capacity, when compared with the individual component MAbs. Furthermore, we show that a MAb with only partial protective capacity but positive effects on the binding of the other BEP component can be used as a valuable constituent of the BEP.


Assuntos
Anticorpos Monoclonais/metabolismo , Antitoxinas/metabolismo , Imunoterapia/métodos , Toxina Tetânica/antagonistas & inibidores , Tétano/prevenção & controle , Animais , Modelos Animais de Doenças , Camundongos , Ligação Proteica , Resultado do Tratamento
12.
PLoS One ; 12(7): e0180551, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28678871

RESUMO

Following infection, the balance between protective immunity and immunopathology often depends on the initial infectious load. Several studies have investigated the effect of infectious dose; however, the mechanism by which infectious dose affects disease outcomes and the development of a protective immune response is not known. The aim of this study was to investigate how the infectious dose modulates the local and systemic humoral and the cellular immune responses during primary ocular chlamydial infection in the guinea pig animal model. Guinea pigs were infected by ocular instillation of a Chlamydophila caviae-containing eye solution in the conjunctival sac in three different doses: 1×102, 1×104, and 1×106 inclusion forming units (IFUs). Ocular pathology, chlamydial clearance, local and systemic C. caviae-specific humoral and cellular immune responses were assessed. All inocula of C. caviae significantly enhanced the local production of C. caviae-specific IgA in tears, but only guinea pigs infected with the higher doses showed significant changes in C. caviae-specific IgA levels in vaginal washes and serum. On complete resolution of infection, the low dose of C. caviae did not alter the ratio of CD4+ and CD8+ cells within guinea pigs' submandibular lymph node (SMLN) lymphocytes while the higher doses increased the percentages of CD4+ and CD8+ cells within the SMLN lymphocytes. A significant negative correlation between pathology intensity and the percentage of CD4+ and CD8+ cells within SMLN lymphocyte pool at selected time points post-infection was recorded for both 1×104, and 1×106 IFU infected guinea pigs. The relevance of the observed dose-dependent differences on the immune response should be further investigated in repeated ocular chlamydial infections.


Assuntos
Infecções por Chlamydophila/imunologia , Infecções por Chlamydophila/microbiologia , Chlamydophila , Imunidade Celular , Imunidade Humoral , Animais , Chlamydophila/patogenicidade , Relação Dose-Resposta Imunológica , Infecções Oculares , Cobaias , Virulência/imunologia
13.
PLoS One ; 12(2): e0172469, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28235050

RESUMO

We demonstrated that a recombinant banana lectin (rBanLec), which structural characteristics and physiological impacts highly resemble those reported for its natural counterparts, binds murine peritoneal macrophages and specifically modulates their functional characteristics. By using rBanLec in concentrations ranging from 1 µg to 10 µg to stimulate resident (RMs) and thioglycollate-elicited (TGMs) peritoneal macrophages from BALB/c and C57BL/6 mice, we have shown that effects of rBanLec stimulation depend on its concentration but also on the functional status of macrophages and their genetic background. rBanLec, in a positive dose-dependent manner, promotes the proliferation of TGMs from both BALB/c and C57BL/6 mice, while its mitogenic influence on RMs is significantly lower (BALB/c mice) or not detectable (C57BL/6 mice). In all peritoneal macrophages, irrespective of their type and genetic background, rBanLec, in a positive dose dependent manner, enhances the secretion of IL-10. rBanLec stimulation of RMs from both BALB/c and C57BL/6 resulted in a positive dose-dependent promotion of proinflammatory phenotype (enhancement of NO production and IL-12 and TNFα secretion, reduction of arginase activity). Positive dose-dependent skewing toward proinflammatory phenotype was also observed in TGMs from C57BL/6 mice. However, the enhancement of rBanLec stimulation promotes skewing of TGMs from BALB/c mice towards anti-inflammatory profile (reduction of NO production and IL-12 secretion, enhancement of arginase activity and TGFß and IL-4 secretion). Moreover, we established that rBanLec binds oligosaccharide structures of TLR2 and CD14 and that blocking of signaling via these receptors significantly impairs the production of TNFα and NO in BALB/c macrophages. Since the outcome of rBanLec stimulation depends on rBanLec concentration as well as on the functional characteristics of its target cells and their genetic background, further studies are needed to investigate its effects under physiological and specific pathological conditions.


Assuntos
Fatores Imunológicos/farmacologia , Interleucina-10/imunologia , Macrófagos Peritoneais/efeitos dos fármacos , Lectinas de Plantas/farmacologia , Tioglicolatos/farmacologia , Animais , Arginase/genética , Arginase/imunologia , Proliferação de Células/efeitos dos fármacos , Relação Dose-Resposta Imunológica , Regulação da Expressão Gênica , Interleucina-10/agonistas , Interleucina-10/genética , Interleucina-12/genética , Interleucina-12/imunologia , Receptores de Lipopolissacarídeos/genética , Receptores de Lipopolissacarídeos/imunologia , Ativação de Macrófagos/efeitos dos fármacos , Macrófagos Peritoneais/citologia , Macrófagos Peritoneais/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Óxido Nítrico/biossíntese , Óxido Nítrico/imunologia , Ligação Proteica , Proteínas Recombinantes/farmacologia , Transdução de Sinais , Receptor 2 Toll-Like/genética , Receptor 2 Toll-Like/imunologia , Fator de Crescimento Transformador beta/genética , Fator de Crescimento Transformador beta/imunologia , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/imunologia
14.
Int J Antimicrob Agents ; 48(6): 732-735, 2016 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-27742207

RESUMO

Nowadays bacterial resistance to known antibiotics is a serious health problem. In order to achieve more efficient treatment, lately there is an effort to find new substances, such as certain biomaterials, that are non-toxic to humans with antibiotic potential. Lignins and lignin-derived compounds have been proposed to be good candidates for use in medicine and health maintenance. In this study, the antibacterial activity of the lignin model polymer dehydrogenate polymer (DHP) in alginate hydrogel (Alg) was studied. The obtained results show that DHP-Alg has strong antimicrobial activity against several bacterial strains and biofilms and does not have a toxic effect on human epithelial cells. These results strongly suggest its application as a wound healing agent or as an adjunct substance for wound treatments.


Assuntos
Alginatos , Anti-Infecciosos/farmacologia , Bactérias/efeitos dos fármacos , Materiais Biocompatíveis , Portadores de Fármacos , Hidrogel de Polietilenoglicol-Dimetacrilato , Lignina/farmacologia , Anti-Infecciosos/toxicidade , Biofilmes/efeitos dos fármacos , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/fisiologia , Ácido Glucurônico , Ácidos Hexurônicos , Humanos , Lignina/toxicidade , Testes de Sensibilidade Microbiana , Ferimentos e Lesões/tratamento farmacológico
15.
PLoS One ; 11(9): e0157875, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27636704

RESUMO

Recent advances in the development of chlamydia vaccines, using live-attenuated or ultraviolet light-inactivated chlamydia, are paving the way for new possibilities to oppose the societal challenges posed by chlamydia-related diseases, such as blinding trachoma. An effective subunit vaccine would mitigate the risks associated with the use of a whole-cell vaccine. Our rationale for the design of an efficient subunit vaccine against Chlamydia trachomatis (Ct) is based on the membrane proteins involved in the initial Ct-host cell contact and on the route of immunization that mimics the natural infection process (i.e., via the ocular mucosa). The first aim of our study was to characterize the specific conjunctival and vaginal immune responses following eye drop immunization in BALB/c mice, using the N-terminal portion of the Ct serovar E polymorphic membrane protein C (N-PmpC) as the subunit vaccine antigen. Second, we aimed to examine the adjuvant properties of the probiotic Lactobacillus rhamnosus (LB) when formulated with N-PmpC. N-PmpC applied alone stimulated the production of N-PmpC- and Ct serovar B-specific antibodies in serum, tears and vaginal washes, whereas the combination with LB significantly enhanced these responses. The N-PmpC/LB combination initiated a T cell response characterized by an elevated percentage of CD25+ T cells and CD8+ effector T cells, enhanced CD4+ T-helper 1 skewing, and increased regulatory T cell responses. Together, these results show that eye drop vaccination with combined use of N-PmpC and a live probiotic LB stimulates specific cellular and humoral immune responses, not only locally in the conjunctiva but also in the vaginal mucosa, which could be a promising approach in Ct vaccine development.


Assuntos
Proteínas de Bactérias/imunologia , Chlamydia trachomatis/imunologia , Lacticaseibacillus rhamnosus , Mucosa Bucal/imunologia , Probióticos , Animais , Feminino , Camundongos , Camundongos Endogâmicos BALB C
16.
Microbes Infect ; 18(4): 254-62, 2016 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-26706818

RESUMO

The aim of this study was to determine whether infectious dose of Chlamydia caviae after repeated infections influences the immunological responses and subsequent clearance of pathogen at the ocular surface of guinea pigs. Animals were infected three times via the conjunctiva at six- and twelve-week intervals by applying either 1 × 10(4) or 1 × 10(6) inclusion-forming units (IFUs) of C. caviae. Ocular pathology, infection course, C. caviae-specific serum IgG levels and their capacity to bind and neutralize infection ex vivo were assessed. Animals infected with 1 × 10(4) IFUs had completely diminished ocular infection and pathology after the 2nd infection with increased levels of C. caviae-specific serum IgG and their effective capacity to bind and neutralize C. caviae. Only partial protection was observed in animals infected with 1 × 10(6) IFUs after the 2nd and 3rd infections. Our findings show that full protection was observed in animals repeatedly infected with the lower dose. The lower dose appeared not to compromise the host immune system, thereby enabling fast clearance of the pathogen and the establishment of competent neutralizing antibodies.


Assuntos
Infecções por Chlamydia/imunologia , Infecções por Chlamydia/microbiologia , Chlamydia/imunologia , Chlamydia/patogenicidade , Infecções Oculares/imunologia , Infecções Oculares/microbiologia , Animais , Anticorpos Antibacterianos/sangue , Anticorpos Neutralizantes/sangue , Infecções por Chlamydia/patologia , Infecções por Chlamydia/prevenção & controle , Modelos Animais de Doenças , Infecções Oculares/patologia , Infecções Oculares/prevenção & controle , Cobaias , Imunoglobulina G/sangue
17.
PLoS One ; 10(12): e0144380, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26656797

RESUMO

Trachoma, caused by the intracellular bacterium Chlamydia trachomatis (Ct), remains the world's leading preventable infectious cause of blindness. Recent attempts to develop effective vaccines rely on modified chlamydial antigen delivery platforms. As the mechanisms engaged in the pathology of the disease are not fully understood, designing a subunit vaccine specific to chlamydial antigens could improve safety for human use. We propose the delivery of chlamydia-specific antigens to the ocular mucosa using particulate carriers, bacterial ghosts (BGs). We therefore characterized humoral and cellular immune responses after conjunctival and subcutaneous immunization with a N-terminal portion (amino acid 1-893) of the chlamydial polymorphic membrane protein C (PmpC) of Ct serovar B, expressed in probiotic Escherichia coli Nissle 1917 bacterial ghosts (EcN BGs) in BALB/c mice. Three immunizations were performed at two-week intervals, and the immune responses were evaluated two weeks after the final immunization in mice. In a guinea pig model of ocular infection animals were immunized in the same manner as the mice, and protection against challenge was assessed two weeks after the last immunization. N-PmpC was successfully expressed within BGs and delivery to the ocular mucosa was well tolerated without signs of inflammation. N-PmpC-specific mucosal IgA levels in tears yielded significantly increased levels in the group immunized via the conjunctiva compared with the subcutaneously immunized mice. Immunization with N-PmpC EcN BGs via both immunization routes prompted the establishment of an N-PmpC-specific IFNγ immune response. Immunization via the conjunctiva resulted in a decrease in intensity of the transitional inflammatory reaction in conjunctiva of challenged guinea pigs compared with subcutaneously and non-immunized animals. The delivery of the chlamydial subunit vaccine to the ocular mucosa using a particulate carrier, such as BGs, induced both humoral and cellular immune responses. Further investigations are needed to improve the immunization scheme and dosage.


Assuntos
Adesinas Bacterianas/imunologia , Chlamydia trachomatis/imunologia , Portadores de Fármacos/química , Olho/imunologia , Mucosa/imunologia , Material Particulado/química , Vacinas de Subunidades Antigênicas/imunologia , Animais , Western Blotting , Proliferação de Células , Túnica Conjuntiva/imunologia , Modelos Animais de Doenças , Epitopos , Escherichia coli/metabolismo , Olho/microbiologia , Olho/patologia , Feminino , Cobaias , Imunização , Imunoglobulina A/metabolismo , Imunoglobulina G/sangue , Injeções Subcutâneas , Interferon gama/metabolismo , Interleucina-4/metabolismo , Camundongos Endogâmicos BALB C , Mucosa/microbiologia , Mucosa/patologia , Proteínas Recombinantes/metabolismo , Baço/patologia , Lágrimas/metabolismo , Tracoma/imunologia , Tracoma/microbiologia , Tracoma/patologia , Tracoma/prevenção & controle
18.
Toxicon ; 103: 135-44, 2015 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-26140841

RESUMO

Antibodies capable to neutralize tetanus toxin (TeNT) are key factors in protection against tetanus disease. Although antibody-based therapeutics for treatment of tetanus exist on the market its production is tedious. Hence, the tetanus-specific antibodies preparation that could be easily produced in large scale in vitro would be beneficial. Monoclonal antibodies (MAbs) are considered for a long time as a reagent of choice, but the core drawback is how to select a MAb that would be safe in providing efficacious protection. In this study we have investigated the parameters crucial for a single MAb to be assigned as protective. Eight murine MAbs were characterized in vitro for their reactivity toward TeNT and assessed in vivo for protectiveness against TeNT intoxication. Correlation of in vitro and in vivo data has revealed that in vitro selection of MAb that is protective in vivo could be performed by a combination of two assays: the measurement of MAb affinity toward TeNT taking Ka 1 × 10(8) M(-1) as a threshold level, and the evaluation of its capability to prevent TeNT-ganglioside interaction. Single MAb could be taken into consideration as a potential therapeutic only if it has a capacity to completely inhibits TeNT-ganglioside complex formation.


Assuntos
Afinidade de Anticorpos , Gangliosídeos/sangue , Antitoxina Tetânica/sangue , Tétano/prevenção & controle , Animais , Anticorpos Monoclonais/sangue , Anticorpos Monoclonais/imunologia , Meia-Vida , Camundongos , Ligação Proteica , Tétano/imunologia , Antitoxina Tetânica/imunologia , Toxina Tetânica/antagonistas & inibidores , Toxina Tetânica/imunologia
19.
J Sci Food Agric ; 95(11): 2271-8, 2015 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-25298065

RESUMO

BACKGROUND: An inductively coupled plasma-optical emission spectrometry method for the speedy simultaneous detection of 19 elements in edible nuts (walnuts: Juglans nigra; almonds: Prunus dulcis; hazelnuts: Corylus avellana; Brazil nuts: Bertholletia excelsa; cashews: Anacardium occidentalle; pistachios: Pistacia vera; and peanuts: Arachis hypogaea) available on the Serbian markets, was optimized and validated through the selection of instrumental parameters and analytical lines free from spectral interference and with the lowest matrix effects. RESULTS: The analysed macro-elements were present in the following descending order: Na > Mg > Ca > K. Of all the trace elements, the tested samples showed the highest content of Fe. The micro-element Se was detected in all the samples of nuts. The toxic elements As, Cd and Pb were either not detected or the contents were below the limit of detection. One-way analysis of variance, Student's t-test, Tukey's HSD post hoc test and hierarchical agglomerative cluster analysis were applied in the statistical analysis of the results. CONCLUSION: Based on the detected content of analysed elements it can be concluded that nuts may be a good additional source of minerals as micronutrients.


Assuntos
Dieta , Espectrometria de Massas/métodos , Minerais/análise , Valor Nutritivo , Nozes/química , Oligoelementos/análise , Anacardiaceae/química , Arachis/química , Bertholletia/química , Humanos , Juglandaceae/química , Prunus dulcis/química , Selênio/análise
20.
Chem Pharm Bull (Tokyo) ; 61(9): 913-9, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23995355

RESUMO

The new kinetically-based spectrophotometric method for the determination of microquantities of ampicillin is proposed in the present paper. Ampicillin degradation in strong alkaline medium was applied for the method development. The reaction rate was monitored at 265 nm. A differential variation of the tangent method was used to process the kinetic data. The method is valid over the 3.49-55.84 µg/mL ampicillin concentration interval with relative standard deviation (RSD) range 7.79-3.20%. The calculated detection limit was determined at 2.58 µg/mL based on the 3.3S0 criterion. The interference effects of some metal ions, anions, amino acids and other molecules were investigated in order to assess the method selectivity. The method was successfully applied to determining the content of ampicillin in commercial pharmaceutical preparations and human urine. The obtained results were in good correlation with the HPLC method results. The newly developed method is simple, inexpensive and efficient for the analysis of a large number of samples at room temperature in a short time.


Assuntos
Ampicilina/análise , Ampicilina/urina , Antibacterianos/análise , Antibacterianos/urina , Humanos , Limite de Detecção , Preparações Farmacêuticas/química , Espectrofotometria/métodos
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