Multicenter phase 1/2 application of adenovirus-specific T cells in high-risk pediatric patients after allogeneic stem cell transplantation

Background

Adenovirus (ADV) reactivation can cause significant morbidity and mortality in children after allogeneic stem cell transplantation. Antiviral drugs can control viremia, but viral clearance requires recovery of cell-mediated immunity.

Evaluation of Essential Trace and Toxic Elements in Biological Samples of Normal and Night Blindness Children of Age Groups 3–7 and 8–12 Years

The most common cause of blindness in developing countries is vitamin A deficiency. The World Health Organization estimates 13.8 million children to have some degree of visual loss related to vitamin A deficiency. The causes of night blindness in children are multifactorial, and particular consideration has been given to childhood nutritional deficiency, which is the most common problem found in underdeveloped countries. Such deficiency can result in physiological and pathological processes that in turn influence biological samples composition. Vitamin and mineral deficiency prevents more than two billion people from achieving their full intellectual and physical potential. This study was designed to compare the levels of Zn, Mg, Ca, K, Na, As, Cd, and Pb in scalp hair, blood, and urine of night blindness children age ranged 3–7 and 8–12 years of both genders, comparing them to sex- and age-matched controls. A microwave-assisted wet acid digestion procedure was developed as a sample pretreatment, for the determination of As, Ca, Cd, K, Pb, Mg, Na, and Zn in biological samples of night blindness children. The proposed method was validated by using conventional wet digestion and certified reference samples of hair, blood, and urine. The concentrations of trace and toxic elements were measured by atomic absorption spectrophotometer prior to microwave-assisted acid digestion. The results of this study showed that the mean values of As, Cd, Na, and Pb were significantly higher in scalp hair, blood, and urine samples of male and female night blindness children than in referents (p?<?0.001), whereas the concentrations of Zn, Ca, K, and Mg were lower in the scalp hair and blood but higher in the urine samples of night blindness children. These data present guidance to clinicians and other professional investigating deficiency of essential mineral elements in biological samples (scalp hair and blood) of night blindness children.     

Simultaneous and sequential co-immobilization of glucose oxidase and catalase onto florisil

The co-immobilization of Aspergillus niger glucose oxidase (GOD) with bovine liver catalase (CAT) onto florisil (magnesium silicate-based porous carrier) was investigated to improve the catalytic efficiency of GOD against H2O2 inactivation. The effect of the amount of bound CAT on the GOD activity was also studied for 12 different initial combinations of GOD and CAT, using simultaneous and sequential coupling. The sequentially co-immobilized GOD-CAT showed a higher efficiency than the simultaneously co-immobilized GOD-CAT in terms of the GOD activity and economic costs. The highest activity was shown by the sequentially co-immobilized GOD-CAT when the initial amounts of GOD and CAT were 10 mg and 5 mg per gram of carrier. The optimum pH, buffer concentration, and temperature for GOD activity for the same co-immobilized GOD-CAT sample were then determined as pH 6.5, 50 mM, and 30 degrees C, respectively. When compared with the individually immobilized GOD, the catalytic activity of the co-immobilized GOD-CAT was 70% higher, plus the reusability was more than two-fold. The storage stability of the co-immobilized GOD-CAT was also found to be higher than that of the free form at both 5 degrees C and 25 degrees C. The increased GOD activity and reusability resulting from the co-immobilization process may have been due to CAT protecting GOD from inactivation by H2O2 and supplying additional O2 to the reaction system.     

DNA extraction from crayfish exoskeleton

Crayfish exoskeleton (CE) samples are generally less invasive and easy to be collected. However, it is difficult to extract DNA from them. This study was intended to investigate CE as a DNA source and design an easy and efficient DNA extraction protocol for polymerase chain reactions. Specific primer pair (PPO-F, PPO-R) was used to amplify extracted DNA from CE, and compared to crayfish tail muscle DNA sample. Moreover, seven microsatellites markers were used to amplify the CE DNA samples set. Since the extracted DNA from CE is suitable for gene amplification, the results present usefulness of CE as an easy and convenient DNA source for PCR-based population genetic research.     

Kinetic properties and storage stability of catalase immobilized on to florisil

The covalent immobilization of bovine liver catalase (CAT) on to florisil via glutaraldehyde was investigated. Optimum immobilization pH and temperature were determined as pH 6.0, 10 degrees C respectively, while the amount of initial CAT per g of carrier and immobilization time was determined as 5 mg g(-1) and 120 min, respectively. The Vmax values for free and immobilized CAT were found to be 1.7 x 10(5) and 2.0 x 10(4) micromol H2O2 min(-1) mg protein(-1), respectively, whereas KM values were 33.3 mM and 1722.0 mM respectively. Operational stability was determined by using a stirred batch-type column reactor. Immobilized CAT retained about 40% of its initial activity after 50 uses. It showed higher storage stability than free CAT at 4 degrees C and 25 degrees C. Its storage stability increased with increasing relative humidity (RH) from 0 to 20% of the medium. The highest storage stability was obtained in 20% RH, however, further increase in RH from 40 to 100% significantly decreased the storage stability.     

Intraflagellar transport delivers tubulin isotypes to sensory cilium middle and distal segments

Sensory cilia are assembled and maintained by kinesin-2-dependent intraflagellar transport (IFT). We investigated whether two Caenorhabditis elegans α- and β-tubulin isotypes, identified through mutants that lack their cilium distal segments, are delivered to their assembly sites by IFT. Mutations in conserved residues in both tubulins destabilize distal singlet microtubules. One isotype, TBB-4, assembles into microtubules at the tips of the axoneme core and distal segments, where the microtubule tip tracker EB1 is found, and localizes all along the cilium, whereas the other, TBA-5, concentrates in distal singlets. IFT assays, fluorescence recovery after photobleaching analysis and modelling indicate that the continual transport of sub-stoichiometric numbers of these tubulin subunits by the IFT machinery can maintain sensory cilia at their steady-state length.     

Seed germination at different temperatures and seedling emergence at different depths of Rhamnus spp

Rhamnus alaternus and R. ludovici-salvatoris, two Mediterranean shrubs with different geographic distributions, have shown important differences in seedling recruitment capacity. The objectives of this work were to determine the ability of these species to germinate seeds under different temperature ranges, as well as the capacity of seedlings to emerge from different burial depths, in order to better understand their regeneration processes. Two different experiments were performed. In the first one, seed germination was studied in Petri dishes and in the dark at different temperature regimes: a) 5–15°C, b) 10–20°C and c) 15–25°C (12h/12h). In the second experiment, seedling emergence capacity from different burial depths (0.5, 2 and 5 cm) was tested. R. ludovici-salvatoris showed a significantly higher final germination rates, a lower dormancy period, and average time response at 10–20°C than at other temperature ranges, although differences were much greater when seeds were subjected to the 5–15°C temperature regime. By contrast, R. alaternus did not show significant differences between treatments (5–15°C and 10–20°C) in germination behavior. Seedling emergence of both species was lower and slower when seeds were buried at 5 cm. However, R. ludovici-salvatoris always showed a lower seedling emergence capacity than R. alaternus at any burial depth. The low ability of R. ludovici-salvatoris to germinate seeds and emerge between 5–15°C, even from shallow depths, is discussed in relation to its low regeneration capacity and declining geographic distribution.     

A retrospective controlled study of thiol disulfide homeostasis as a novel marker in Crimean Congo hemorrhagic fever

Objectives: Crimean Congo hemorrhagic fever (CCHF) is the second most common hemorrhagic fever worldwide. This study aimed to evaluate the oxidant–antioxidant balance of patients with CCHF by detecting dynamic thiol disulfide homeostasis (TDH), which is a novel oxidative stress marker, and other molecules, including paraoxonase (PON), arylesterase (ARES), ceruloplasmin (CLP), myeloperoxidase (MPO), and catalase.

Methods: This retrospective, cross-sectional, controlled study, which involved patients with CCHF and healthy volunteers, measured dynamic TDH using a novel automated method developed by Erel.

Results: We recruited 69 adult patients with CCHF (31 females, 38 males, median age 46 years). The case fatality rate was 1.49% (1/69). Increased disulfide/native thiol and disulfide/total thiol ratios, decreased total antioxidant status (TAS), and increased total oxidant status (TOS) were found in patients with CCHF. TAS, PON, and ARES values were found to be positively correlated with both native and total thiol levels, whereas TOS and CLP were negatively correlated with both, at a significant level. MPO activity was similar in both groups.

Discussion: This is the first study in the literature to evaluate dynamic TDH in CCHF. TDH shifts to the oxidative side in patients with CCHF, leading to an increase in TOS.