Evaluation of persistent COVID and SARS-CoV-2 reinfection in a cohort of patients on the island of Gran Canaria, Spain / Evaluación de la COVID-19 persistente y la reinfección por SARS-CoV-2 en una cohorte de pacientes en la isla de Gran Canaria, España

Introduction This study analyzed the impact of patients’ age, sex, vaccination, immunosuppressive treatment, and previous comorbidities on the risk of developing persistent COVID-19 or SARS-CoV-2 virus reinfection. Method Population-based observational retrospective study of a cohort of 110,726 patients aged 12 years or older, who were diagnosed with COVID-19 between June 1st, 2021 and February 28th, 2022 in the island of Gran Canaria. Results 340 patients suffered reinfection. The combination of advanced age, female sex and lack of complete or incomplete vaccination against COVID-19 was strongly predictive of reinfection (p<0.05). In the 188 patients who developed persistent COVID-19, the persistence of symptoms was more frequent in adult patients, women, and patients with a diagnosis of asthma. Complete vaccination was associated with a lower risk of reinfection ([OR] 0.05, 95%CI 0.04–0.07; p<0.05) and of developing persistent COVID-19 ([OR] 0.07, 95%CI 0.05–0.10; p<0.05). None of the patients with reinfection or persistent COVID-19 died during the period of the study Conclusions This study confirmed the link between age, sex, asthma and risk of persistent COVID-19. It was not possible to define the patient's comorbidities as a factor that influences the development of reinfection, but its association with age, sex, type of vaccine and hypertension was demonstrated. Higher vaccination coverage was associated with a lower risk of persistent COVID-19 or SARS-CoV-2 reinfection (AU)

Introducción Se analizó el impacto de la edad, el sexo, la vacunación, el tratamiento inmunosupresor y las comorbilidades previas del paciente sobre la condición de riesgo de desarrollar COVID-19 persistente o reinfección por el virus del SARS-CoV-2. Método Estudio retrospectivo observacional de base poblacional en una cohorte de 110.726 pacientes de 12 o más años de edad diagnosticados de COVID-19 entre el 1 de junio de 2021 y el 28 de febrero de 2022 en la isla de Gran Canaria. Resultados Trescientos cuarenta pacientes sufrieron reinfección por COVID-19. La combinación de edad avanzada, sexo femenino y falta de vacunación completa o incompleta contra la COVID-19 fue fuertemente predictiva de reinfección (p<0,05). En los 188 pacientes que desarrollaron COVID-19 persistente, la persistencia de síntomas fue más frecuente en pacientes en edad adulta, mujeres y pacientes con diagnóstico de asma. La vacunación completa se asoció con un menor riesgo de reinfección ([OR] 0,05, IC 95% 0,04-0,07; p <0,05) y de desarrollar COVID-19 persistente ([OR] 0,07, IC 95% 0,05-0,10; p <0,05). Ninguno de los pacientes con reinfección o COVID-19 persistente falleció durante el período del estudio. Conclusiones Este estudio confirmó el vínculo entre la edad, el sexo, el asma y el riesgo de COVID-19 persistente. No se pudo definir las comorbilidades del paciente como factor que influye en el desarrollo de reinfección, pero sí se demostró su asociación con edad, sexo e hipertensión arterial. Una mayor cobertura de vacunación se asoció a un menor riesgo de COVID-19 persistente o reinfección por SARS-CoV-2 (AU)

Evaluation of persistent COVID and SARS-CoV-2 reinfection in a cohort of patients on the island of Gran Canaria, Spain.

INTRODUCTION: This study analyzed the impact of patients' age, sex, vaccination, immunosuppressive treatment, and previous comorbidities on the risk of developing persistent COVID-19 or SARS-CoV-2 virus reinfection. METHOD: Population-based observational retrospective study of a cohort of 110,726 patients aged 12 years or older, who were diagnosed with COVID-19 between June 1st, 2021 and February 28th, 2022 in the island of Gran Canaria. RESULTS: 340 patients suffered reinfection. The combination of advanced age, female sex and lack of complete or incomplete vaccination against COVID-19 was strongly predictive of reinfection (p<0.05). In the 188 patients who developed persistent COVID-19, the persistence of symptoms was more frequent in adult patients, women, and patients with a diagnosis of asthma. Complete vaccination was associated with a lower risk of reinfection ([OR] 0.05, 95%CI 0.04-0.07; p<0.05) and of developing persistent COVID-19 ([OR] 0.07, 95%CI 0.05-0.10; p<0.05). None of the patients with reinfection or persistent COVID-19 died during the period of the study. CONCLUSIONS: This study confirmed the link between age, sex, asthma and risk of persistent COVID-19. It was not possible to define the patient's comorbidities as a factor that influences the development of reinfection, but its association with age, sex, type of vaccine and hypertension was demonstrated. Higher vaccination coverage was associated with a lower risk of persistent COVID-19 or SARS-CoV-2 reinfection.

Stabilization of porous chitosan improves the performance of its association with platelet-rich plasma as a composite scaffold.

This study offers innovative perspectives for optimizing of scaffolds based on correlation structure-function aimed the regenerative medicine. Thus, we evaluated in vitro performance of stabilized porous chitosan (SPCHTs) associated with activated platelet-rich plasma (aP-PRP) as a composite scaffold for the proliferation and osteogenic differentiation of human adipose-derived mesenchymal stem cells (h-AdMSCs). The porous structure of chitosan (PCHT) was prepared similarly to solid sponges by controlled freezing (-20 °C) and lyophilization of a 3% (w/v) chitosan solution. Stabilization was performed by treating the PCHT with sodium hydroxide (TNaOH), an ethanol series (TEtOH) or by crosslinking with tripolyphosphate (CTPP). The aP-PRP was obtained from the controlled centrifugation of whole blood and activated with autologous serum and calcium. Imaging of the structures showed fibrin networks inside and on the surface of SPCHTs as a consequence of electrostatic interactions. SPCHTs were non-cytotoxic, and the porosity, pore size and Young's modulus were approximately 96%, 145 µm and 1.5 MPa for TNaOH and TEtOH and 94%, 110 µm and 1.8 MPa for CTPP, respectively. Stabilization maintained the integrity of the SPCHTs for at least 10 days of cultivation. SPCHTs showed controlled release of the growth factors TGF-ß1 and PDGF-AB. Although generating different patterns, all of the stabilization treatments improved the proliferation of seeded h-AdMSCs on the composite scaffold compared to aP-PRP alone, and differentiation of the composite scaffold treated with TEtOH was significantly higher than for non-stabilized PCHT. We conclude that the composite scaffolds improved the in vitro performance of PRP and have potential in regenerative medicine.

Targeting and retention of HPV16 E7 to the endoplasmic reticulum enhances immune tumour protection.

The endoplasmic reticulum (ER) is where the major histocompatibility complex (MHC) class I molecules are loaded with epitopes to cause an immune cellular response. Most of the protein antigens are degraded in the cytoplasm to amino acids and few epitopes reach the ER. Antigen targeting of this organelle by Calreticulin (CRT) fusion avoids this degradation and enhances the immune response. We constructed a recombinant adenovirus to express the E7 antigen with an ER-targeting signal peptide (SP) plus an ER retention signal (KDEL sequence). In cell-culture experiments we demonstrated that this new E7 antigen, SP-E7-KDEL, targeted the ER. Infection of mice with this recombinant adenovirus that expresses SP-E7-KDEL showed interferon induction and tumour-protection response, similar to that provided by an adenovirus expressing the E7 antigen fused to CRT. This work demonstrated that just by adding a SP and the KDEL sequence, antigens can be targeted and retained in the ER with a consequent enhancement of immune response and tumour protection. These results will have significant clinical applications.

Effect of high-oxygen and high-carbon-dioxide atmospheres on strawberry flavor and other quality traits.

The effect of high-oxygen atmospheres on strawberry flavor was studied. Strawberry fruits (Fragariax ananassa Duch. cv. Camarosa) were stored at 8 degrees C in four different atmospheres: air, 5% O(2)/20% CO(2), 80% O(2)/20% CO(2), and 90% O(2)/10% CO(2). Changes in several quality parameters were evaluated. Atmospheres combining high O(2) and high CO(2) were the most effective in preventing fungal growth and enhancing strawberry firmness. Other quality parameters such as color, titrable acidity, sugars and organic acids distribution, off-flavor development, and aroma were only mildly affected by superatmospheric O(2) levels. After one week of storage, unexpected high contents of off-flavor related compounds were found in the 80% O(2)/20% CO(2) and 90% O(2)/10% CO(2) atmospheres. Evidence of an altered ester biosynthesis was also found in fruits stored under these high-O(2) atmospheres. Data obtained suggest that stress induced by high CO(2) and stress induced by high O(2) have an additive effect on strawberry flavor alteration.

Cytosolic aldolase is a ripening related enzyme in strawberry fruits (Fragaria x ananassa).

Two aldolase isoenzymes have been isolated from ripe strawberry fruits (Fragaria x ananassa cv. Camarosa and Elsanta) and partially purified by DEAE anion exchange and Sephacryl size exclusion chromatography. The isoenzymes were identified as class I cytosol and plastid aldolase on the basis of their chromatographic behavior on DEAE-cellulose columns, native molecular weight, pH optimum pattern, Km value for D-fructose-1,6-bisphosphate, tendency to be inactivated by lower pH values and SDS-PAGE subunit determination of 40 and 38 kDa, respectively. Total aldolase activity and distribution of both aldolase isoenzymes was also investigated at different stages of strawberry fruit ripening. Strawberries in the green and white ripening stage showed the same ratio of the two isoenzymes as green leaves with 15 and 8% cytosol aldolase activity, respectively. During strawberry fruit development the overall total aldolase activity decreased until the pink ripening stage and then increased due to a rise of cytosol aldolase yielding up to 75% in red strawberries. A cDNA putatively encoding the cytosolic form of aldolase in strawberry was cloned during the course of this study. Both microarray and RNA gel blot analyses showed that the cytosolic aldolase gene expression is induced during ripening as detected for the cytosolic aldolase enzyme. We suggest that induction of the cytosolic aldolase both at the levels of transcription and translation might be part of a ripening related stress response in the receptacle tissue.

Electrochemical oxidation of cisatracurium on carbon paste electrode and its analytical applications.

The electrochemical oxidation of cisatracurium was investigated by cyclic voltammetry and differential pulse voltammetry at a carbon paste electrode and the experimental parameters have been optimized in order to obtain the optimum analytical signal. A differential pulse voltammetric method with carbon paste electrode is described for the determination of cisatracurium with detection limit of 0.38 mug/ml and quantitation limit of 1.26 mug/ml. The proposed method was applied to determine the content of cisatracurium in human urine and human serum, obtaining accurate and precise results.

Relationship between clinical and biologic variables and chloramphenicol pharmacokinetic parameters in pediatric patients with sepsis.

OBJECTIVE: To evaluate the influence of several clinical and biologic factors on the disposition kinetics of oral chloramphenicol in pediatric patients and to determine the usefulness of this information to predict chloramphenicol serum concentrations. STUDY DESIGN: The clinical, biologic, and pharmacokinetic data of 30 consecutive pediatric patients diagnosed with sepsis and admitted to a tertiary care center were analyzed retrospectively. The patients were randomly assigned to a study group and a validation group. The model was developed by a three-step approach involving Bayesian estimation of pharmacokinetic parameters, selection of covariates by principal component analysis, and final selection by stepwise multiple linear regression. The model was tested in the study group and compared with a general population model using a prediction error analysis. RESULTS: Regression analysis revealed that weight, albumin, and white blood cell (WBC) count were the most important determinants for chloramphenicol distribution volume, whereas age, WBC count, and serum creatinine were the most important determinants for chloramphenicol clearance. The performance of the constructed population model improved significantly in terms of both bias and precision compared with the general model when tested in the validation group. CONCLUSIONS: Clinical and biologic factors may significantly influence chloramphenicol's disposition in pediatric patients with sepsis and therefore should be considered in programming dosage regimens.

Lipoxygenase and hydroperoxide lyase activities in ripening strawberry fruits.

The enzymes lipoxygenase and hydroperoxide lyase have been identified in strawberry (Fragariax ananassa Duch.) var. Camarosa. Their subcellular localization, substrate preference, and product specificity were determined in mature strawberry fruits. The activity of both enzymes was located mainly in the microsomal fraction. Linolenic acid was the preferred substrate for strawberry lipoxygenase, forming 13- and 9-hydroperoxides of this acid in the proportion 70:30. The strawberry hydroperoxide lyase cleaves 13-hydroperoxide of linoleic (13% relative activity) and linolenic (100% relative activity) acids to form hexanal and (3Z)-hexenal, respectively. Both enzyme activities and endogenous content of volatile aldehydes formed by sequential action of lipoxygenase-hydroperoxide lyase were evaluated during strawberry development and ripening. A sequential enzymatic pathway for the formation of green odor compounds in strawberry is proposed.

Biosynthesis of 4-hydroxy-2,5-dimethyl-3(2H)-furanone and derivatives in in vitro grown strawberries.

The biosynthesis of 2,5-dimethyl-4-hydroxy-3(2H)-furanone (Furaneol) and its methyl ether and glucoside derivatives has been studied in strawberries. An in vitro system was used for growing this fruit, showing that the presence in the incubation medium of sucrose or hydroxyquinoline hemisulfate has no effect on the bioformation of these compounds. Strawberries in vitro grown showed an increase in furanone content with time, especially between the second and fourth days, to the same extent as field-grown fruits but at a higher rate. Among the precursors added to the incubation medium, D-fructose gave rise to an increase in furaneol and its glucoside derivative of 42. 6% and 26.3%, respectively. D-fructose 6-phosphate seems to be the precursor of furaneol in strawberries since, when present in the incubation medium, it produced an average increase of 125% in all furanones contents with respect to control fruits.

Effects of ozone treatment on postharvest strawberry quality.

The effect of ozone treatment on the postharvest quality of strawberry was evaluated. Strawberry fruits (Fragaria x ananassa Duch. cv. Camarosa) were stored at 2 degrees C in an atmosphere containing ozone (0.35 ppm). After 3 days at 2 degrees C, fruits were moved to 20 degrees C to mimic retail conditions (shelf life). The changes in several quality parameters such as fungal decay, color, sugar and acids distribution, and aroma were evaluated during the strawberries' shelf life. Ozone treatment was ineffective in preventing fungal decay in strawberries after 4 days at 20 degrees C. Significant differences in sugars and ascorbic acid content were found in ozone-treated strawberries. At the end of cold storage, the vitamin C content of ozonated strawberries was 3 times that of control fruits. A detrimental effect of ozone treatment on strawberry aroma was observed, with a 40% reduced emission of volatile esters in ozonated fruits.

Characterization of three potato lipoxygenases with distinct enzymatic activities and different organ-specific and wound-regulated expression patterns.

Lipoxygenases are ubiquitous enzymes in eukaryotes. In plants, lipoxygenases are involved in the synthesis of the hormone jasmonic acid that regulates plant responses to wounding and, in addition, is an inducer of tuberization in potato. We have isolated potato lipoxygenase cDNA clones. From their deduced amino acid sequences, three distinct classes are defined (Lox1, Lox2, and Lox3). They are encoded in gene families that display organ-specific expression, lox1 being expressed mostly in tubers and roots, lox2 in leaves, and lox3 in leaves and roots. Consistent with their organ-specific expression pattern, Lox1 expressed in bacteria preferentially uses as substrate linoleic acid, abundant in membrane lipids of tubers, whereas linolenic acid, prevalent in leaves, is the preferred substrate for the other two classes of lipoxygenase. Analyses on reaction products of the enzymes expressed in bacteria reveal that Lox1 primarily produces 9- hydroperoxides. In contrast, the jasmonic acid precursor, 13-hydroperoxylinolenic acid, is the major product of the action of Lox2 and Lox3 on linolenic acid. Upon wounding, the levels of Lox2 and Lox3 transcripts rise markedly in leaves. While Lox3 mRNA accumulation peaks as early as 30 min after wounding, Lox2 shows a steady increase over a 24-h time course, suggesting different roles for these lipoxygenase isoforms in the synthesis of the plant hormone jasmonic acid.

Immunophenotypic analysis of lymphocytes and myeloma cells in patients with multiple myeloma.

The immunological phenotypes of lymphocytes and myeloma cells in 48 patients with multiple myeloma (MM) were analyzed using a panel of monoclonal antibodies (mAbs). Myeloma cells were positive for OKT10, BL3, PCA1 and BA2. In a few cases, they were also positive for the B cell-associated antigens J5, B1 and I2. Eight of 48 cases had more than 15% J5-positive lymphocytes, and some lymphocytes in MM expressed plasma cell-associated antigens (PCA1, BL3, OKT10), suggesting a possible clonal involvement. These observations demonstrate the heterogeneity of surface antigen expression of myeloma cells and suggest that BL3, PCA1, BA2 and J5 may be useful mAbs for purging myeloma cells from bone marrow for autologous transplantation.

A computational method for approximate evaluation of ionization constants from potentiometric data.

A general equation is given for the evaluation of proton dissociation constants from potentiometric data. The method is applicable to monobasic acids, and also polybasic acids, where the successive dissociation steps may or may not overlap. A computer program was employed for the calculations. The method was tested by evaluation of the constants for 22 acids, from monobasic to octobasic, with satisfactory agreement with literature data.

Discrepancy between flow cytometric analyses of CALLA using two monoclonal antibodies.

Both the J5 and BA-3 monoclonal antibodies are considered to be specific for epitopes on the common acute lymphoblastic leukemia antigen (CALLA). Flow-cytometric analyses of three cell lines and one normal bone marrow sample using these antibodies as CALLA markers demonstrated that J5-labeled cells were always brighter than those labeled with BA-3, and that the ratio of their fluorescence intensities varied widely in the different systems. Furthermore, one of the lines, RPMI 8226, while positive for J5, appeared to be negative when labeled with BA-3, except for a slight displacement of the fluorescence distribution relative to the control. A possible explanation for the observed results is that the BA-3 binding epitope or epitopes on CALLA may vary in their number and/or accessibility to the antibody. These observations suggest that the use of a single monoclonal antibody to detect a cell surface antigen may be misleading, particularly when a negative result is obtained.

Extraction and spectrophotometric determination of titanium(IV) with alizarin and fluoride.

The characteristics of the mixed-ligand titanium(IV)-fluoride-alizarin complex, including the optimum conditions of formation and extraction into methyl isobutyl ketone, are described. A simple and sensitive procedure for spectrophotometric determination of titanium has been developed. At pH 9.5-10.3 titanium reacts with alizarin in the presence of fluoride to form a red-violet complex that is completely extractable into methyl isobutyl ketone, and has its absorption maximum at 513 nm. The molar absorptivity at 513 nm is 7.0 x 10(4)l.mole(-1).cm(-1). Beer's law is obeyed up to 22 mug of titanium in 30 ml of solution. The method has been used for the determination of titanium in an oxide mixture and aluminium alloy samples.

Salicylaldehyde-1-phthalazinohydrazone as an analytical reagent.

The synthesis, characteristics and analytical properties of salicylaldehyde-1-phthalazinohydrazone are described. Spectral characteristics, pK values, the effect of oxidizing and reducing agents, resistance to hydrolysis, and reactions with common cations are reported.

Factors modifying the synergistic toxicity of deoxycytidine in combination with thymidine plus 5-fluorouracil in HeLa cells.

We reported previously that deoxycytidine (CdR) enhances the cytotoxic effects of the drug combination thymidine (TdR) plus 5-fluorouracil (FUra) against HeLa S-3 cells. We have now examined the relationships between the concentration of CdR and its cytotoxic and cytokinetic effects, and have also investigated the role of certain other components of the culture medium in this phenomenon. Cell survival was determined by a colony-forming assay; cytokinetic effects were monitored by flow cytometry. In the initial experiments, cells were grown in Ham's F12 medium and exposed for 22 hr to 4 mM TdR, 0 X 025 mM FUra, and dCyd ranging from 1 microM to 4 X 0 mM. The individual drugs were at most only slightly toxic under these conditions; for TdR plus FUra, the survival decreased to 50% (in 5% FCS), and in the three-drug combination it varied from 8% at 1 microM CdR to 28% at 0 X 10 mM and back to a low of 3% at 4 X 0 mM CdR. Results from flow cytometry appeared correlated with the survival data, in that cells accumulated in the S phase to a greater extent in the region around 0 X 10 mM CdR than at higher or lower concentrations. When cells were exposed to the drugs in MEM medium in place of F12, their sensitivity to FUra and the TdR-FUra combination was enhanced, although the additional synergistic effect of CdR was reduced. We found that hypoxanthine, present in F12 but not in MEM, was the principal compound responsible for the observed differences between media.

Effects of cytosine arabinoside and daunorubicin on survival and cell cycle progression of Chinese hamster ovary cells.

Kinetic and cytotoxic effects of cytosine arabinoside (Ara-C) and daunorubicin (DNR) on exponentially growing Chinese hamster ovary (CHO) cells were measured by flow cytometry and by a colony-forming assay, respectively. With Ara-C alone, increasing drug concentrations between 10(-7) M and 10(-4) M, for up to 27 hr, were associated with increased inhibition of cell progression through the S phase. Even at the very toxic concentration of 10(-4) M, however, cells were able to enter and progress slowly through S. DNR, which appears to enter these cells relatively slowly, was highly toxic even at 2 X 10(-7) M. It decreased the rate of progression through S phase and caused cells to accumulate in G2, except at the highest concentration (2 X 10(-5) M, at which progression was inhibited throughout the cycle. Simultaneous exposure of the cells to Ara-C and DNR yielded cell cycle distributions similar to those of the former drug alone. When cells were exposed to a non-lethal dose of Ara-C and to a dose of DNR which was lethal to a fraction of the cell population (or conversely), either simultaneously or separated by a drug-free interval, small, but in some cases significant, drug interactions were observed. These effects were not caused by drug-induced redistribution of cells within the cell cycle, but may have been related to the effects of the non-lethal drug on DNA synthesis rate.