[Three-month functional prognosis of patients hospitalised due to acute ischaemic stroke in Aragon: Rregional analysis of the impact of COVID-19]. / Pronóstico funcional a 3 meses de pacientes hospitalizados por ictus isquémico agudo en Aragón: análisis regional de la repercusión de la COVID-19.

Introduction: The COVID-19 pandemic has had an impact on ischaemic stroke management, with a reported decrease in hospital admissions, and even disruptions in healthcare and increased in-hospital mortality. However, there is a lack of evidence on the impact of the pandemic on functional prognosis. The aim of this study is to analyse the effect of the COVID-19 pandemic on the 3-month functional outcomes of patients hospitalised due to acute ischaemic stroke in Aragon (Spain). Materil and methods: We reviewed the data of all patients admitted due to ischaemic stroke to any hospital in our regional healthcare system between 30 December 2019 and 3 May 2020. We compared modified Rankin Scale scores and mortality at 3 months in patients hospitalised before and after the declaration of a state of emergency due to the COVID-19 pandemic. Results: In total, 318 patients with acute ischaemic stroke met our inclusion criteria. No differences were observed between periods in global or specific characteristics, with the exception of a higher proportion of patients older than 80 years during the first period (42.2% vs. 29.0%, P=.028). In the comparative analysis, we found no significant differences in mortality (12.3 vs. 7.9, P=.465) or in the proportion of patients with modified Rankin Scale scores ≤ 2 (57.7% vs. 57.1%, P=.425) at 3 months. Conclusion: To our knowledge, this is the first study to analyse the impact of COVID-19 pandemic on the 3-month functional outcomes of patients with ischaemic stroke. In our region, there has been no increase in rates of mortality or disability at 3 months in patients admitted due to ischaemic stroke during the pandemic.

Three-month functional prognosis of patients hospitalised due to acute ischaemic stroke in Aragon: regional analysis of the impact of COVID-19.

INTRODUCTION: The COVID-19 pandemic has had an impact on ischaemic stroke management, with a reported decrease in hospital admissions, and even disruptions in healthcare and increased in-hospital mortality. However, there is a lack of evidence on the impact of the pandemic on functional prognosis. The aim of this study is to analyse the effect of the COVID-19 pandemic on the 3-month functional outcomes of patients hospitalised due to acute ischaemic stroke in Aragon (Spain). METHODS: We reviewed the data of all patients admitted due to ischaemic stroke to any hospital in our regional healthcare system between 30 December 2019 and 3 May 2020. We compared modified Rankin Scale scores and mortality at 3 months in patients hospitalised before and after the declaration of a state of emergency due to the COVID-19 pandemic. RESULTS: In total, 318 patients with acute ischaemic stroke met our inclusion criteria. No differences were observed between periods in global or specific characteristics, with the exception of a higher proportion of patients older than 80 years during the first period (42.2% vs 29.0%, P = .028). In the comparative analysis, we found no significant differences in mortality (12.3 vs 7.9, P = .465) or in the proportion of patients with modified Rankin Scale scores ≤ 2 (57.7% vs 57.1%, P = .425) at 3 months. CONCLUSION: To our knowledge, this is the first study to analyse the impact of COVID-19 pandemic on the 3-month functional outcomes of patients with ischaemic stroke. In our region, there has been no increase in rates of mortality or disability at 3 months in patients admitted due to ischaemic stroke during the pandemic.

Modulation of allergic response by gene-environment interaction: olive pollen allergy.

This article summarizes the most important advances of recent years in the field of gene-environment interaction in allergic response. It specifically examines sensitization to olive pollen as an example of one of the main causes of allergic disease in the Mediterranean area. The presence of at least 20 proteins with allergic activity has been demonstrated in olive pollen, and 10 of these have been characterized (Ole e 1 to Ole e 10). Ole e 1, which is considered to be the majority allergen (causing sensitization in more than 70% of patients), has been the subject of many studies looking for risk factors and ways to protect against sensitization. Markers of the major histocompatibility complex and other genetic loci associated with the allergic response have been analyzed using population-based, family-based, and functional approaches, which have revealed the involvement of genetic regulation in this type of response. Furthermore, evaluation of environmental factors and their relationship with genetic factors is essential when attempting to understand this type of disease. In this review, we provide examples of how exposure to high doses of olive pollen allergen in a specific genetic context can trigger different allergic conditions (from asthma to nonresponse). We stress the importance of evaluating these factors in order to modulate this response correctly.

Expression on human eosinophils of CD148: a membrane tyrosine phosphatase. Implications in the effector function of eosinophils.

The role of protein tyrosine phosphatases (PTP) is crucial in regulating the phosphorylation status of cells. CD148 is a recently described membrane-type PTP. In this study, we have demonstrated that this molecule is expressed on human eosinophils and eosinophilic cell line EoL-3. Interestingly, our data also showed that this molecule acts as a transduction molecule on these cells. Thus, the crosslinking of CD148 was able to induce the degranulation and the induction of superoxide anion generation. By using specific inhibitor and by western blotting, we have shown that tyrosine kinase activation is involved in this transduction pathway. In addition, we have shown the presence of a serine/threonine kinase activity associated with CD148. In conclusion, the activation capacity of CD148 on eosinophils suggests a potential role of this molecule on inflammatory diseases, such as allergic and parasitic diseases, associated with eosinophilia.

Olive (Olea europea) pollen allergens--II. Isolation and characterization of two major antigens.

A dialyzed extract of olive (Olea europea) pollen was fractionated by anion exchange chromatography on DEAE-Sepharose CL-6B using a discontinuous gradient of ammonium bicarbonate. The most important protein allergen was obtained from the 0.3 M fraction after gel filtration on Sephadex G-100 and separation by lentil-lectin Sepharose-4B. The major allergen of olive pollen was contained in the effluent and was designated Olea Antigen I. This material inhibited the RAST activity of 15 patients' sera that were tested. Analytical IEF demonstrated a major band at pH 5.3 and two minor ones at pH 5.6 and 5.0. When these were run into SDS-polyacrylamide gel electrophoresis in a second dimension, all were separated into two bands of mol. wt 17 and 19 K. A second protein, which is the next most important allergen, Olea Antigen II, was obtained from the 0.5 M fraction by chromatofocusing in a 4-7 pH range followed by filtration on Bio-gel P-30. Olea Antigen II had a mol. wt of 8 K as assessed by SDS-PAGE. IEF analysis displayed one main band at pH 3.6 and two minor bands at pH 3.8 and 4.0, respectively. OL-1, an anti-Olea europea monoclonal antibody (MAb) previously reported by us Lauzurica et al. (1988) reacted with the 17 and 19 K antigens from the crude extract and with Olea Antigen I but not with Olea Antigen II.

Effects of dietary fat saturation on fatty acid composition and gene transcription in different tissues of Iberian pigs.

The effect of two diets, respectively enriched with SFA (S) and PUFA (P), on FA tissue composition and gene expression was studied in fattened Iberian pigs. The FA composition of adipose, muscular and liver tissues was affected by dietary treatment. S group showed higher MUFA and MUFA/SFA ratio and lower PUFA and n-6/n-3 ratio than P group in all analyzed tissues. In muscle and liver the extracted lipids were separated into neutral lipids and polar lipid fractions which showed significantly different responses to the dietary treatment, especially in liver where no significant effect of diet was observed in NL fraction. The expression of six candidate genes related to lipogenesis and FA oxidation was analyzed by qPCR. In liver, stearoyl CoA desaturase (SCD), acetyl CoA carboxylase alpha (ACACA) and malic enzyme 1 (ME1) genes showed higher expression in S group. SCD, ACACA, ME1, and fatty acid synthase (FASN) gene expression levels showed a wide variation across the tested tissues, with much higher expression levels observed in adipose tissue than other tissues. Tissue FA profile and gene expression results support the deposition of dietary FA, the lipogenic effect of dietary saturated fat in liver and the employment of saturated dietary fat for endogenous synthesis of MUFA in all the analyzed tissues.

Molecular cloning and characterization of an IgE-reactive protein from Anisakis simplex: Ani s 1.

Ingestion of the parasitic nematode Anisakis simplex in undercooked fish can cause severe allergic reactions in some individuals. Using pooled human sera from sensitized patients we have probed an expression library for A. simplex antigens. One positive clone was found to encode a full length 21 kDa protein with strong homology to nematode troponins. The recombinant protein was expressed as a GST-fusion protein and found by immunoblot analysis to react with sera from 20% of allergic patients. The presence of functional EF-hand Ca(2+) binding motifs was demonstrated by gel-shift analysis.

Immunogold probes for light and electron microscopic localization of Ole e I in several Oleaceae pollens.

We investigated the immunolocalization of the olive major allergen Ole e I and Ole e I-like proteins in pollen from several Oleaceae species [olive (Olea europaea), ash (Fraxinus excelsior), privet (Ligustrum vulgaris), lilac (Syringa vulgare), and forsythia (Forsythia suspensa)]. Crossreactions among different pollens were found in enzyme immunoassays. For immunolocalization with light microscopy we used the silver enhancement technique with three monoclonal antibodies (1D8, 10H1, and 16G2) that recognize three different epitopes of the allergen Ole e I. Our findings show that the silver enhancement technique is very useful when several antibodies are to be used for rapid screening of different materials. MAb 10H1 gave the most precise results and was selected for further immunolocalization studies with transmission electron microscopy. The epitope recognized by this MAb was localized exclusively in the endoplasmic reticulum in olive pollen. In lilac, privet, and ash pollen, most of the reactivity was also seen in the endoplasmic reticulum; however, the 10H1 epitope was not detected in forsythia pollen.

The effect of human pregnancy serum on the synthesis and action of interleukin-1.

Supernatants from mixed lymphocyte reactions performed in the presence of pregnancy serum (PS) or normal human serum (NHS) were tested for interleukin-1 (IL-1) activity in a thymocyte assay. A lower response was obtained with PS-containing supernatants, suggesting that PS exerts its suppressive effect by inhibiting IL-1 synthesis or by blocking its mode of action. IL-1 activity generated by human peripheral blood mononuclear cells (PBMC) stimulated with LPS and cultured in the presence of PS or NHS was similar. However, when PS was present, the proliferation of thymocytes co-mitogenically stimulated with PHA and exogenous IL-1 was diminished compared with the control (NHS), exhibiting its maximum inhibition when PS was added at the highest concentration and at the beginning of the incubation period. The suppressive effect of PS in the thymocyte assay disappeared with the combined action of exogenous IL-1 and IL-2. The response of T lymphocytes allogeneically stimulated was decreased in the presence of PS even when IL-1 was added; the recovery of proliferation being strictly dependent on the presence of the appropriate dose of exogenous IL-2. These results indicate that PS was interfering with IL-1 activity by rendering IL-2-producer cells unable to synthesize IL-2 and subsequently to proliferate.

Human pregnancy serum inhibits proliferation of T8-depleted cells and their interleukin-2 synthesis in mixed lymphocyte cultures.

T8-depleted and unfractionated T lymphocytes allogeneically stimulated and cultured in the presence of pregnancy sera exhibit an inhibition of cellular proliferation and interleukin-2 synthesis, respectively. Unfractionated T cells show a decrease in their cytotoxicity in the presence of these sera. The inhibition of cytotoxicity could be due to the deficit of IL-2 observed since if exogenous IL-2 is added to the cultures T4/LEU3a-depleted allogeneically stimulated cells reach the same degree of cytotoxicity whether cultured in normal human serum or pregnancy serum. A possible mechanism to explain the inhibition of mixed lymphocyte cultures by pregnancy serum could therefore be decrease of cellular proliferation of the T8-depleted subpopulation with a decrease in IL-2 synthesis, implying an inhibition of cytotoxic effector cells.

Study of apoptosis in human lymphocytes by toxic substances implicated in toxic oil syndrome.

Toxic Oil Syndrome is a multisystemic disease that occurred in epidemic proportions in Spain in 1981 caused by the ingestion of rapeseed oil denatured with aniline. Several data implicate T cells in the pathogenesis of the disease. We evaluated the mechanisms of cytotoxicity in human lymphocytes of TOS-related products: aniline, 3-(N-phenylamino)-1,2-propanediol and its mono- and di-oleyl esters and eosinophilia myalgia-related product such as 3-(phenylamino)-L-alanine, which is chemically similar to 3-(N-phenylamino)-1,2-propanediol, and has been found in manufactured L-tryptophan. Our results show that only di-oleyl ester of 3-(N-phenylamino)-1,2-propanediol induces apoptosis in human lymphocytes, in a concentration and time-dependent way, confirmed by morphology, expression of phosphatidylserine in membrane and analysis of DNA degradation.

Immunological basis of toxic oil syndrome (TOS).

The toxic oil syndrome (TOS), a multisystemic disease, that occurred in Spain in 1981, was caused by the ingestion of rapeseed oil denatured with 2% aniline. Due to the clinical course of the disease, immunopathological mechanisms have been suspected but a direct connection was never demonstrated. To analyse this possibility, we determined several immunological parameters in the sera of patients with TOS and without the disease, using a case-control design: total immunoglobulins, IgG and IgE antibodies against different toxic agents (oleylanilide, aniline, linoleyl-anilide, and 3-phenylaminopropane-1-2-diol), autoantibodies, cytokines (IL-4, IL-6, TNF, GM-CSF) and soluble receptors (sCD23 and sIL-2R). We detected high levels of sIL-2R in TOS patients compared to controls (P < 0.0001). A higher levels of sCD23 and IgE were also found. In addition, the response to oleyl-anilide of peripheral blood lymphocytes from TOS patients was studied and a significant proliferative response in 30% of TOS patients versus 5% controls was observed. Our data support the implication of the immune system in the acute phase of TOS, with a possible activation of T-cells and release of cytokines, that could explain some of the clinical findings in this phase of the disease.

Cytokine mRNA expression in lung tissue from toxic oil syndrome patients: a TH2 immunological mechanism.

In 1981, an epidemic occurred in Spain, toxic oil syndrome (TOS), in people who consumed rapeseed oil denatured with 2% aniline, and it was one of the largest intoxication epidemics ever recorded. In 1989, a similar disease, eosinophilia-myalgia syndrome (EMS) was reported in the USA and was associated with the ingestion of L-tryptophan. The pathologic findings in TOS showed primary endothelial injury, with cell proliferation and perivascular inflammatory infiltrates. Immunologic mechanisms have presumably been operative in the pathogenesis and perpetuation of TOS. Our previous findings pointed to a T-cell activation during acute phase of the disease. In order to analyze which T-cell subset is involved on TOS, we have developed an mRNA extraction procedure from paraffin-embedded lung tissues in patients with pulmonary involvement. We analyzed mRNA expression from different cytokines (IL-1, IL-2, IL-4, IL-5, IFN-gamma, GM-CSF) and CD25 (interleukin 2 receptor) and CD23 (low affinity IgE receptor), using RT-PCR technique. In lung tissues from these patients a T-cell activation was observed. We found a significant increase in Th1 (P = 0.006) and Th2 (P = 0.003) cytokine profile in TOS patients with respect to controls. The increment in TH2 response with respect to TH1 is significant (P = 0.03) in TOS lung specimens. Non-significant differences were obtained in other cytokines and receptors studied as IL-1, CD25, CD23 and GM-CSF. Data presented in this paper are the first clear evidence that an immunological mechanism is directly implicated in this illness.

Experimental bovine hypodermosis in Spain.

An experimental survey was carried out in western Spain to investigate both the chronobiology of Hypoderma spp. and the immunoresponse of their bovine hosts. This study was initiated with a new system of obtaining Hypoderma spp. larvae directly from their host, including the eclosion of adults from their pupae, infestation under natural but controlled conditions, and confirmation of the resulting infection. This survey was carried out over 2 cattle grub seasons; it was possible to infest and reinfest the experimental animals and to monitor them by both parasitological methods and by an enzyme-linked immunosorbent assay. This method permitted the evaluation of the development of anti-Hypoderma antibodies during the experiment. The experimental design also enabled us to establish the period of detectable H. lineatum infection to be from December until the end of April with the largest number of warbles observed during March and April. After a pupal period of < 30 d, adults were seen in April and May. Hypoderma bovis (de Geer) showed a delay of 2 m.o. relative to H. lineatum (de Villiers). This study reports a completed biological life cycle of Hypoderma spp. under controlled conditions in both natural and experimental environments.

Immunohistochemical study of the local immune response to Fasciola hepatica in primarily and secondarily infected goats.

The distribution of CD2, CD4, CD8, gamma/delta T-lymphocytes, B-cells and IgG lambda-light chain (lambda-IgG) containing cells were analysed in the inflammatory infiltrate associated to hepatic lesions and gallbladder (HL), and in hepatic lymph nodes (HLN) of goats primarily and secondarily infected with Fasciola hepatica. In the HL, CD2 and CD8 T-cells were more numerous (p=0.01) in secondarily rather than in primarily infected goats, whereas CD4 T-lymphocytes were less numerous than CD8 and showed no significant change in both groups. The ratio CD4/CD8 was 0.66 and 0.39 for primarily and secondarily infected goats, respectively. In contrast, in the HLN, CD4 were more numerous than CD8 T-cells, the ratio CD4/CD8 was 2.0 in control, 1.5 and 1.3 in primary and secondary infections, respectively. Gamma/delta T-lymphocytes were scarce in the HL and moderate in the HLN of both primarily and secondarily infected animals. B-cells (IgM+, lambda-IgG+ or CD79+) varied from scarce or moderate in the HL to abundant in the HLN, where CD79+-cells were mainly located in lymphoid follicles and IgM and IgG in plasma-cells of the medullary cords, suggesting an intense local humoral immune response. However, this response did not prevent the hepatic damage in secondarily infected goats, in which hepatic lesions were more severe than in primarily infected ones.

Comparison of erythromycin and oxytetracycline for the treatment of ovine footrot.

A microbiological study of 25 cases of ovine footrot was performed. Cultures belonging to Dichelobacter nodosus were isolated in 48% of the sampled animals. The sensitivity of the 99 strict anaerobic bacterial isolates to 5 antibiotics (penicillin G, amoxycillin, spiramycin, erythromycin and oxytetracycline) was studied. The percentage of resistant cultures was in all cases higher than 30%. The efficacy of erythromycin and oxytetracycline in the treatment of ovine footrot was studied. To conduct this test, an intramuscular injection was applied, of one antimicrobial or the other, at the beginning of the treatment. The tolerance of animals to the antimicrobials, the success rate of treatment and the severity of lameness were evaluated. The percentage of animals cured within 15 days was around 75%. In contrast, only 44% improvement was achieved in the lameness. No differences were found between the two antimicrobials in the above indices.

Effectiveness of different adjuvants in stimulating Dichelobacter nodosus antibody in sheep vaccinated against ovine footrot.

This research consists of an evaluation of the effectiveness of different substances administered as adjuvants in the stimulation of humoral immune response induced by the vaccine composed of strains A1, A2 and C of Dichelobacter nodosus. To do this, a total of 120 Merino sheep were vaccinated and revaccinated. These sheep were selected from a farm located in the region of Extremadura (Spain), and they were divided into 12 groups of 10 animals each. An additional group with 10 sheep was used as control. The immune response (titre of antibodies) was determined by agglutination tests and ELISA. The most pronounced immune response was obtained by the use of Freund's incomplete adjuvant and aluminium hydroxide as adjuvants.

Rapid chromatographic determination of caseins in milk with photometric and fluorimetric detection using a hydrophobic monolithic column.

Reverse-phase liquid chromatographic methods using a hydrophobic C18 monolithic column and on-line photometric and fluorimetric detection for the determination of the major casein (CN) proteins in milk are presented. The separation of αs1-CN, αs2-CN, ß-CN and κ-CN was achieved in only five minutes. Fluorimetric detection enabled better analytical results than photometric detection. Thus, the dynamic ranges of the calibration graphs and detection limits obtained using fluorimetric detection were (mgmL(-)(1)): αs1-CN (0.74-10.0, 0.22), αs2-CN (0.15-10.0, 0.045), ß-CN (0.68-10.0, 0.20) and κ-CN (0.21-10.0, 0.06). The analytical features of the photometric method, which does not allow the quantification of ß-casein, were (mgmL(-)(1)): αs1-CN (1.5-9.0, 0.45), αs2-CN (1.4-10.0, 0.43) and κ-CN (0.4-9.0, 0.12). Precision data, expressed as relative standard deviation, ranged between 0.6% and 5.3% for the fluorimetric method and between 2.4% and 6.2% for the photometric method. Both methods were applied to the analysis of three different milk samples, obtaining recoveries in the ranges of 86.6-103.2% and 92.0-106.5% using fluorimetric and photometric detection, respectively.