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1.
Photochem Photobiol Sci ; 22(3): 535-547, 2023 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-36378410

RESUMEN

Helicobacter pylori gastric infections are among the most diffused worldwide, suffering from a rising rate of antibiotic resistance. In this context, some of the authors have previously designed an ingestible device in the form of a luminous capsule to perform antibacterial photodynamic inactivation in the stomach. In this study, the light-emitting capsules were tested to verify the safety of use prior to perform clinical efficacy studies. First, laboratory tests measured the capsule temperature while in function and verified its chemical resistance in conditions mimicking the gastric and gut environments. Second, safety tests in a healthy minipig model were designed and completed, to verify both the capsule integrity and the absence of side effects, associated with its illumination and transit throughout the gastrointestinal tract. To this aim, a capsule administration protocol was defined considering a total of 6 animals with n = 2 treated with 8 capsules, n = 2 treated with 16 capsules and n = 2 controls with no capsule administration. Endoscopies were performed in sedated conditions before-after every capsule administration. Biopsies were taken from the corpus and antrum regions, while the gastric cavity temperature was monitored during illumination. The bench tests confirmed a very good chemical resistance and a moderate (about 3 °C) heating of the capsules. The animal trials showed no significant effects on the gastric wall tissues, both visually and histologically, accompanied with overall good animal tolerance to the treatment. The integrity of the administered capsules was verified as well. These encouraging results pose the basis for the definition of successive trials at the clinical level.


Asunto(s)
Antibacterianos , Fototerapia , Animales , Porcinos , Porcinos Enanos , Seguridad de Equipos , Antibacterianos/farmacología
2.
Future Microbiol ; 14: 1199-1205, 2019 09.
Artículo en Inglés | MEDLINE | ID: mdl-31625444

RESUMEN

Aim: The objective of this study was to investigate the possible synergy between doxycycline and photodynamic therapy against Helicobacter pylori and to evaluate the possible side effects on adenocarcinoma gastric cells with and without protoporphyrin IX. Materials & methods: Three H. pylori strains (ATCC 700392, 43504 and 49503) were grown on solid medium either with, or without, doxycycline at subinhibitory concentrations, and irradiated for 10, 20 and 30 minutes with a 400 nm-peaked light source. The phototoxicity tests on AGS cells were evaluated by MTT assay. Results: The photodynamic therapy and doxycycline combination showed an antibacterial synergistic effect with no significant toxicities. Conclusion: The synergistic treatment could be considered as an interesting therapeutic option.


Asunto(s)
Antibacterianos/farmacología , Doxiciclina/farmacología , Helicobacter pylori/efectos de los fármacos , Fotoquimioterapia/métodos , Protoporfirinas/farmacología , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Supervivencia Celular/efectos de la radiación , Dermatitis Fototóxica , Sinergismo Farmacológico , Mucosa Gástrica/citología , Mucosa Gástrica/efectos de los fármacos , Mucosa Gástrica/efectos de la radiación , Infecciones por Helicobacter/tratamiento farmacológico , Infecciones por Helicobacter/radioterapia , Humanos , Pruebas de Sensibilidad Microbiana , Fotoquimioterapia/efectos adversos
3.
Sci Rep ; 9(1): 2064, 2019 02 14.
Artículo en Inglés | MEDLINE | ID: mdl-30765716

RESUMEN

Therapeutic drug and immunogenicity monitoring (TDIM) is increasingly proposed to guide therapy with biologics, characterised by high inter-individual variability of their blood levels, to permit objective decisions for the management of non-responders and reduce unnecessary interventions with these expensive treatments. However, TDIM has not yet entered clinical practice partly because of uncertainties regarding the accuracy and precision of enzyme-linked immunosorbent assays (ELISA). Here we report the characterisation of a novel surface plasmon resonance (SPR)-based TDIM, applied to the measurement of serum concentrations of infliximab, an antibody against tumour necrosis factor α (anti-TNFα), and anti-infliximab antibodies. SPR has the obvious advantages of directly detecting and measuring serum antibodies in minutes, avoiding the long incubation/separation/washing/detection steps of the methods proposed so far, reducing complexity and variability. Moreover, drug and anti-drug antibodies can be measured simultaneously. This new method was validated for sensitivity and reproducibility, and showed cost-effectiveness over commercial ELISA kits. This method may be applied to other biotherapeutics. These data pave the way for the development of SPR-based point-of-care devices for rapid on-site analysis.


Asunto(s)
Anticuerpos Monoclonales/sangre , Bioensayo/métodos , Monitoreo de Drogas/métodos , Ensayo de Inmunoadsorción Enzimática/métodos , Humanos , Infliximab/sangre , Reproducibilidad de los Resultados , Resonancia por Plasmón de Superficie/métodos , Factor de Necrosis Tumoral alfa/inmunología
4.
J Photochem Photobiol B ; 186: 107-115, 2018 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-30029036

RESUMEN

Since many years it has been acknowledged that some bacterial species, among which H. pylori, P. aeruginosa, P. acnes accumulate endogenous photosensitizers (PS) in the form of porphyrins. This makes antibacterial photodynamic therapy (PDT) easier to perform due to the possible avoidance of external PS. In this study, we focus on gastric infections associated with the presence of Helicobacter pylori (H. pylori), known to accumulate and release both protoporphyrin IX (PPIX) and coproporphyrins. PDT versus H. pylori can be carried out by modified endoscopes or by new ingestible luminous devices under development. In both cases of in vitro and in vivo applications, either for therapy (PDT) or diagnosis, scientific literature lacks studies on the possible side-effects of light treatments on the surrounding tissues. To this aim we evaluated in vitro side-effects due to a possible intrinsic photosensitivity of gastric mucosa or to a photosensitization by the PS released from the bacterium itself. Photo-toxicity studies were conducted on the AGS cell line (ATCC® CRL-1739™), commonly used as a model for the stomach mucosa tissue, considering PPIX as the photosensitizing agent. After first evaluations of PPIX dark toxicity, its uptake and accumulation sites, photo-toxicity tests were conducted using a LED light source peaked at 400 nm, by varying both PPIX concentration (50 nM - 2 µM) and light dose in the range 0.6-13 J/cm2, representing different treatment procedures found in literature. The oxidative stress consequent to irradiation was investigated both in terms of ROS production and assessment of the activity of enzymes involved in ROS-related biological mechanisms. A significant phototoxic effect was found only for PPIX concentration > 100 nM for all tested light doses. This indicates that the evaluated photo-treatments do not cause side effects even with the sensitization due to PPIX released by the bacteria.


Asunto(s)
Mucosa Gástrica/efectos de los fármacos , Luz , Fármacos Fotosensibilizantes/farmacología , Apoptosis/efectos de los fármacos , Apoptosis/efectos de la radiación , Línea Celular , Supervivencia Celular/efectos de los fármacos , Supervivencia Celular/efectos de la radiación , Mucosa Gástrica/efectos de la radiación , Infecciones por Helicobacter/tratamiento farmacológico , Infecciones por Helicobacter/microbiología , Infecciones por Helicobacter/patología , Helicobacter pylori/efectos de los fármacos , Helicobacter pylori/efectos de la radiación , Humanos , Microscopía Confocal , Estrés Oxidativo/efectos de los fármacos , Estrés Oxidativo/efectos de la radiación , Fotoquimioterapia/efectos adversos , Fármacos Fotosensibilizantes/uso terapéutico , Protoporfirinas/metabolismo , Protoporfirinas/farmacología , Protoporfirinas/uso terapéutico , Especies Reactivas de Oxígeno/metabolismo
5.
Am J Physiol Endocrinol Metab ; 292(1): E272-80, 2007 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-16940473

RESUMEN

Macrophage migration inhibitory factor (MIF) is an important proinflammatory cytokine involved in regulation of macrophage function. In addition, MIF may also play a role in murine and human reproduction. Although both first trimester trophoblast and decidua express MIF, the regulation and functional significance of this cytokine during human placental development remains unclear. We assessed MIF expression throughout normal human placental development, as well as in in vitro (chorionic villous explants) and in vivo (high altitude placentae) models of human placental hypoxia. Dimethyloxalylglycine (DMOG), which stabilizes hypoxia inducible factor-1 under normoxic conditions, was also used to mimic the effects of hypoxia on MIF expression. Quantitative real-time PCR and Western blot analysis showed high MIF protein and mRNA expression at 7-10 wk and lower levels at 11-12 wk until term. Exposure of villous explants to 3% O(2) resulted in increased MIF expression and secretion relative to standard conditions (20% O(2)). DMOG treatment under 20% O(2) increased MIF expression. In situ hybridization and immunohistochemistry showed elevated MIF expression in low oxygen-induced extravillous trophoblast cells. Finally, a significant increase in MIF transcript was observed in placental tissues from high-altitude pregnancies. Hence, three experimental models of placental hypoxia (early gestation, DMOG treatment, and high altitude) converge in stimulating increased MIF, supporting the conclusion that placental-derived MIF is an oxygen-responsive cytokine highly expressed in physiological in vivo and in in vitro low oxygen conditions.


Asunto(s)
Factores Inhibidores de la Migración de Macrófagos/metabolismo , Oxígeno/fisiología , Placenta/metabolismo , Adulto , Altitud , Vellosidades Coriónicas/metabolismo , Femenino , Expresión Génica , Humanos , Técnicas de Cultivo de Órganos , Placentación , Embarazo
6.
Infect Immun ; 71(11): 6664-7, 2003 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-14573693

RESUMEN

Interleukin-4 (IL-4) and IL-4delta2 mRNA gastric expression was evaluated in healthy subjects and patients who did not have ulcers but were infected with Helicobacter pylori with or without the cag pathogenicity island (cag PAI). IL-4 mRNA was physiologically expressed by gastric epithelium and negatively influenced by H. pylori. Also, nonepithelial cells in the lamina propria of H. pylori-infected patients expressed IL-4 mRNA, whereas IL-4delta2 mRNA was found only in cag PAI-negative patients. Thus, gastric IL-4 takes part in the local immune response to H. pylori.


Asunto(s)
Mucosa Gástrica/inmunología , Infecciones por Helicobacter/inmunología , Helicobacter pylori/patogenicidad , Interleucina-4/genética , ARN Mensajero/análisis , Adulto , Femenino , Helicobacter pylori/inmunología , Humanos , Masculino , Isoformas de Proteínas , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Virulencia
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