Performance of a New Rapid Point-of-Care Test for Infliximab Levels in Patients with Inflammatory Bowel Disease: A Comparison to ELISA.

BACKGROUND: Therapeutic drug monitoring of infliximab levels in patients with inflammatory bowel disease (IBD) optimizes patients' treatment. The reference technique is based on enzyme-linked immunosorbent assay (ELISA) although point of care (POC) assays are being developed. AIMS: To assess the performance of a new rapid immunochromatographic POC assay (Promonitor Quick IFX) compared with ELISA technique to measure infliximab levels in patients with IBD. METHODS: A prospective, observational, unicentric study was performed on capillary blood samples from patients with IBD before infliximab infusion (trough levels). Infliximab levels and anti-infliximab antibodies were measured using the ELISA technique (Promonitor IFX) and the POC assay. Correlation between both techniques was assessed by Pearson's coefficient. Quantitative differences were evaluated by Bland-Altman analysis. Samples were stratified according to infliximab therapeutic ranges (< 3 µg/mL, 3-8 µg/mL, and > 8 µg/mL). RESULTS: A total of 135 experimental samples were assessed. Infliximab levels showed a high correlation between POC and ELISA tests (r = 0.84, P < 0.001). The mean difference between tests was 1.46 µg/mL (P < 0.001), being minimal for concentrations < 8 µg/mL. POC and ELISA assays showed an overall concordance of 87.4%. Most samples were in the same therapeutic range, which lead to equivalent therapeutic decisions. POC and ELISA assays detected the presence of anti-infliximab antibodies in 2.2% and 3.7% of the samples, respectively. CONCLUSIONS: POC assay results in blood samples from patients with IBD were comparable to those obtained with the reference ELISA technique. The POC assay could be considered for routine testing based on its ease of use and rapidity.

Cryptochromes are the dominant photoreceptors mediating heliotropic responses of Arabidopsis inflorescences.

Inflorescence movements in response to natural gradients of sunlight are frequently observed in the plant kingdom and are suggested to contribute to reproductive success. Although the physiological and molecular bases of light-mediated tropisms in vegetative organs have been thoroughly investigated, the mechanisms that control inflorescence orientation in response to light gradients under natural conditions are not well understood. In this work, we have used a combination of laboratory and field experiments to investigate light-mediated re-orientation of Arabidopsis thaliana inflorescences. We show that inflorescence phototropism is promoted by photons in the UV and blue spectral range (≤500 nm) and depends on multiple photoreceptor families. Experiments under controlled conditions show that UVR8 is the main photoreceptor mediating the phototropic response to narrowband UV-B radiation, and phototropins and cryptochromes control the response to narrowband blue light. Interestingly, whereas phototropins mediate bending in response to low irradiances of blue, cryptochromes are the principal photoreceptors acting at high irradiances. Moreover, phototropins negatively regulate the action of cryptochromes at high irradiances of blue light. Experiments under natural field conditions demonstrate that cryptochromes are the principal photoreceptors acting in the promotion of the heliotropic response of inflorescences under full sunlight.

Immediate effects of diamond burr debridement in patients with spontaneous chronic corneal epithelial defects, light and electron microscopic evaluation.

PURPOSE: To evaluate immediate effects of diamond burr debridement (DBD) on the cornea of canine patients diagnosed with spontaneous chronic corneal epithelial defects (SCCEDs). ANIMALS STUDIED: Eight client owned dogs with SCCEDs. METHODS: Nine eyes from eight dogs with SCCEDs underwent superficial keratectomy (SK). The ulcerated area was divided into quadrants with a 300-micron restricted depth knife. Two of four quadrants underwent DBD for 40-60 s. A SK followed immediately. One burred section and one nonburred section were fixed with formaldehyde 10% and underwent light microscopy (LM). The remaining quadrants from five eyes were fixed with glutaraldehyde 2.5% and underwent transmission electron microscopy (TEM). Masked pathologists evaluated the samples. A student's paired t-test was used to analyze the data. RESULTS: With LM all nonburred samples had a superficial stromal hyaline acellular zone (HAZ), seven of the burred samples had an intermittent HAZ and in two burred samples this zone was absent. The HAZ thickness of burred samples (1.062 ± 0.664 µm) was significantly thinner than that of the nonburred samples (4.309 ± 1.348 µm) (P < 0.0001). Transmission electron microscopy showed an absence of basement membrane and the presence of an amorphous, fine fibrillar material in the superficial stroma in nonburred samples. This material was intermittent or absent in burred samples. CONCLUSION: DBD significantly reduces the superficial stromal HAZ in SCCEDs. A reduction of its thickness may be responsible for the healing rates reported with DBD.

Conjunctival squamous cell carcinoma in a reindeer (Rangifer tarandus tarandus).

An 8-year-old female adult reindeer (Rangifer tarandus tarandus) was referred to the Veterinary Hospital of Madrid for evaluation of a conjunctival mass on the left eye which had been present for about 2 months. A surgical excision was performed and biopsy material submitted for light microscopic evaluation which confirmed the diagnosis of conjuctival squamous cell carcinoma. Nuclear p53 immunolabeling was found in 52% of the neoplastic cells. Follow-up examination at 12 months postsurgery did not reveal recurrence of this neoplasm. Conjunctival squamous cell carcinoma has not been reported previously in reindeer and seems to have similar characteristics to the one existing in bovine species.

Morphological and ultrastructural characterization of neurospheres spontaneously generated in the culture from sheep ovarian cortical cells.

Neurospheres (NS) derived from adult stem cells of non-neural tissues represent a promising source of neural stem cells (NSCs) and neural progenitor cells (NPCs) for autologous cell therapy. Knowing the fine structure of NS cells is essential for characterizing them during differentiation or oncogenic transformation. NS are generated by culturing ovarian cortical cells (OCCs) under specific conditions. To establish whether these OCCs exhibited a similar morphophenotype as those from the central nervous system (CNS) reported in the literature, sheep OCCs were cultured for 21 days to generate NS. Expression levels of pluripotency (Nanog, octamer-binding transcription factor 4 [Oct4], and SRY-box transcription factor 2 [Sox2]) and NSCs/NPCs (nestin, paired box 6 [Pax6], and p75 neurotrophin receptor [P75NTR]) transcripts were analyzed by quantitative reverse transcription-polymerase chain reaction (qRT-PCR), the NSC/NPC antigens were immunolocalized, and structural and ultrastructural analyses were performed in OCC-NS on Days 10, 15, and 21 in culture. Spheroids expressed transcripts and antigens of pluripotency as well as NSCs/NPCs. Cells were arranged into an inner core, with frequent apoptotic and degenerative events, and a peripheral epithelial-like cover with abundant cytoplasmic organelles, apical microvilli, and filament bundles of cytoskeleton elements. Adherens junctions and apical tight and lateral loose interdigitations were found in peripheral cells that eventually lost apical-basal polarization, which might indicate their disengaging/aggregating from/to the NS. We can conclude that OCC-NS shares the most structural and ultrastructural characteristics with CNS-NS.

Characterisation of corneal impression cytology in dogs and its application in the diagnosis of keratoconjunctivitis sicca.

OBJECTIVE: Determine morphological and morphometric parameters of corneal epithelium in dogs, and determine the cellular alterations that occur in canine keratoconjunctivitis sicca (KCS) using impression cytology. STUDY ANIMALS: 60 dogs divided into two groups: dogs with Schirmer tear test (STT) at least 15 mm/minute and absence of ocular disease, and dogs with STT less than 15 mm/minute and clinical signs of KCS. PROCEDURES: Impression cytology was used to collect corneal samples. The percentage of eyes with cell changes, the number of such cells and the percentage of cells with structural alterations in each group were determined. The possible correlation between corneal epithelium alterations and decreased tear production was evaluated. RESULTS: A significant positive correlation existed between STT and the area of the cytoplasm and nucleus of corneal cells. A significant negative correlation was found between STT ​​and the nucleus/cytoplasm ratio, and the presence of cellular changes. A significant difference existed between the numbers of pyknotic nuclei, being higher among animals with all stages of KCS. CONCLUSION: Corneal impression cytology can be used to assess the corneal epithelium in healthy eyes and eyes with KCS, demonstrating its usefulness as a diagnostic tool especially in mild and early cases.

Spheroids Spontaneously Generated In Vitro from Sheep Ovarian Cortical Cells Contain Integrating Cells That Exhibit Hallmarks of Neural Stem/Progenitor Cells.

Cell spheroids are inducible or spontaneously generated cell aggregates produced in vitro that can provide a valuable model for developmental biology, stem cell biology, and cancer therapy research. This investigation aimed to define the cellular identity of spheroids spontaneously generated in vitro from sheep ovarian cortical cells cultured under specific serum-free conditions. Spheroids were characterized during 21 days of culture by morphometric evaluation, detection of alkaline phosphatase (AP) activity, gene expression analyses of stemness transcription factors and several lineage markers, immunolocalization analyses, as well as assessment of self-renewal and differentiation potential. Cell aggregation, evidenced from day 3 of culture onward, resulted in efficient generation of 65-75 spheroids for every 500,000 cells seeded. The spheroids reached maximum diameter (187 ± 15.9 µm) during the second week of culture and exhibited AP activity. Sox2, Oct4, and Nanog were expressed throughout the culture period, with upregulation of Sox2. Neural lineage specification genes (eg, nestin, vimentin, Pax6, and p75NTR) were expressed from day 10 onward at levels above that of Oct4, Nanog and those for endoderm [alpha-fetoprotein (AFP)], and mesoderm (brachyury) specification. Neural stem cell (NSC)/neural progenitor cell (NPC) markers, nestin, Pax6, p75NTR, and vimentin, were extensively localized in cells on day 10, 15 (44.75% ± 5.84%; 93.54% ± 1.35%; 78.90% ± 4.80%; 73.82% ± 3.40%, respectively), and 21 (49.98% ± 5.30%; 91.84% ± 1.9%; 76.74% ± 11.0%; 95.80% ± 3.60%, respectively). Spheroid cell self-renewal was evidenced by cell proliferation and the generation of new spheroids during two consecutive expansion periods. Culture of spheroid cells under differentiation conditions gave rise to cells showing immunolocalization of the neuron-specific antigen NeuN and the astroglial antigen GFAP (glial fibrillary acidic protein). Our results indicate that spheroids spontaneously generated in this culture system were comprised of cells with molecular characteristics of NSC/NPC that can self-renew and differentiate into neurons and glia, supporting the identity of spheroids as neurospheres.

Nuevos tratamientos psicológicos en tabaquismo: las intervenciones a distancia / New psychological treatments in tobacco dependence: new non face-to-face smoking cessation interventions

La prevalencia de tabaquismo sigue siendo muy elevada en nuestro país. Sin embargo, no todos los usuarios tienen un acceso fácil a los tratamientos para dejar de fumar. El avance de las nuevas tecnologías ha permitido el desarrollo de programas de tratamiento de tabaquismo a distancia, tanto a través de intervenciones telefónicas o quitlines, como de Internet. La Unidad de Tabaquismo del Hospital Carlos III ha incorporado a su rutina asistencial este tipo de tratamientos. El presente trabajo tiene como objetivo revisar las publicaciones sobre estos nuevos tratamientos de tabaquismo a distancia, así como ver su aplicabilidad clínica en un hospital público con experiencia previa en tratamientos presenciales

Smoking prevalence is still elevated in our country. Neverthless most of smokers do not have an easy acces to smoking cessation treatment. Development of non face-to-face programs though quitlines and internet has been possible thanks to new technologies advance. The objective of this paper is to review the research about new non face-to-face smoking cessation treatments and to analyse its clinic feasibility in a public hospital with previous experience in actual treatments