Age and parasitism status of Tuta absoluta eggs alter the foraging responses of the predator Nabis pseudoferus

We studied the effect of Trichogramma brassicae Bezdenko (Hymenoptera: Trichogrammatidae) parasitism of Tuta absoluta Meyrick (Lepidoptera: Gelechiidae) eggs on the foraging behavior of Nabis pseudoferus Remane (Hemiptera: Nabidae) feeding on T. absoluta eggs of different ages. Older parasitized eggs (48–72 h old) were rejected after probing with the probosis. A separate experiment showed that probing did not harm developing parasitoids. Bugs greatly reduced their feeding times on older parasitized eggs and spent more time grooming and probing eggs and leaves. Predators exhibited a type III functional response to 24 h-old eggs, but a type II response to older eggs, whether parasitized or not. 24 h-old eggs suffered higher attack rates, and required less handling time than older eggs. Although the predatory efficiency of N. pseudoferus was reduced in the presence of parasitized eggs, the partial refuge enjoyed by T. brassicae in older eggs may permit an additive impact on the pest, and should facilitate the joint augmentation of both species against T. absoluta.

     

Optimization of the Endotoxin Removal Performance of Solid-Phase Conjugated S3E3 Antimicrobial Peptide Using Response Surface Methodology

S3E3 is a new variant of S3 antimicrobial peptide (AMP) derived from factor C of horseshoe hemolymph and features a high binding affinity for endotoxin. In this research, site-specific conjugated S3E3 AMP onto Sepharose 6% solid phase support (S3E3-S-Sepharose) was applied for endotoxin removal from protein solutions. The bovine serum albumin (BSA) was chosen as a protein model due to its acidic-sticky nature interfering with the endotoxin removal process. The batch process parameters including, endotoxin concentration, pH, and ionic strength of the sample were optimized by response surface methodology to reach maximum endotoxin binding capacity and protein recovery. The predicted optimal conditions for enhanced endotoxin removal performance were as follows: pH 4.5, 25 mM NaCl, and 68,500 EU/ml endotoxin leading to a maximum endotoxin binding capacity of 3.114?×?10⁺⁶ EU/ml of resin and a 95.89% protein recovery. S3E3-S-Sepharose could be applied as an efficient endotoxin removal affinity chromatography matrix at downstream processes of recombinant therapeutics due to its high capacity and protein recovery.

     

Immobilization of recombinant nanobiofiber CS3 fimbriae onto alginate beads for improvement of cadmium biosorption

A cell surface display system with metalbinding properties was previously developed using CS3 fimbriae, which are hollow tubes 20 nm-thick and 2 nm in diameter. In this study, hybrid CS3 pili were separated from recombinant Escherichia coli and entrapped in calcium alginate gel beads in order to improve their stabilization and also adsorption of heavy metals. The surface morphology of the gel beads containing pili was investigated by scanning electron microscopy (SEM). Immunofluorescence microscopy was employed to confirm the attachment of nanobiofibers to the alginate beads. The effects of three variables (sodium alginate concentration, protein to alginate mass ratio, and bead size) at two levels each on Cd²⁺ biosorption efficiency were investigated by full factorial experimental design. A second-order polynomial equation modeled the design space for the process response of cadmium removal capacity. The optimal values of the factors were obtained as follows: 1% sodium alginate concentration, 0.25 protein to alginate mass ratio, and a 6 mm bead size. Under these conditions, Cd²⁺ was adsorbed at 45.45 mg/g to the nanobiofiber. The results indicate that the immobilized recombinant hybrid CS3 pili may be an appropriate biosorbent for removal of heavy metals from polluted aquatic environments.     

Evaluation of mammalian codon usage of fimH in DNA vaccine design

Uropathogenic Escherichia coli (UPEC) bacteria are the principal cause of urinary tract infections (UTI). Because these bacteria propagate intracellularly, the cellular immune response is an important factor in UTIs. Therefore, we designed a genetic construct to induce a cellular immune response. In order to develop a genetic construct that induces strong cellular immunity against this pathogen, we used the fimH synthetic gene according to mammalian codon usage, and the gene expression was compared with wild type codon usage. Initially, we designed two constructs, pVAX/fimH mam and pVAX/fimH wt, which contain mammalian and wild type codon usage, respectively. The Cos-7 cell line was transfected separately with a complex of pVAX/fimH mam-ExGene 500 poly cationic polymer and pVAX/fimH wt-ExGene 500 poly cationic polymer. Expression of the fimH gene in both constructs in COS7 cells was confirmed by RT-PCR, SDS-PAGE, and Western blotting. Both of the pVAX/fimH cassettes expressed inserted fimH genes (mam and wt) in Cos-7 cells. Our results suggest that codon optimization successfully expressed the fimH gene because the fimH gene with mammalian codon usage is compatible with the eukaryotic expression system. Therefore, mammalian codon usage could be appropriate in a pVAX/fimH construct as a DNA vaccine.     

Optimization of human Th17 cell differentiation in vitro<Emphasis Type="Italic">:</Emphasis> evaluating different polarizing factors

Regarding discrepancies that exist among different studies which have tried to clarify critical factors in human Th17 cell differentiation, the aim of this study was to identify the best condition for human Th17 differentiation and to clarify the possible role of TGF-β in differentiation of these cells. Naïve CD4⁺ T cells were isolated from cord blood samples and cultured either in X-VIVO 15 serum-free medium or RPMI 1640 containing 10% FBS. Purified cells were treated with different combinations of polarizing cytokines (TGF-β, IL-1β, IL-6, IL-23 and IL-21) followed by analysis of the expression of characteristic genes and their relevant cytokines by real-time quantitative RT-PCR and ELISA method, respectively. Our data indicate that a combination of TGF-β plus IL-6 and IL-23 cytokines in X-VIVO 15 serum-free medium could be applied as the best condition for developing human Th17 cells in compare with other studied cytokine treatments. It is shown that TGF-β could be considered as a positive regulator for human Th17 cell differentiation only if applied in average concentrations. Interestingly, polarizing treatments in absence of TGF-β, induced double-secreting Th17 cells which co-express IL-17 and IFN-γ whereas polarization in presence of TGF-β-induced single-secreting (only IL-17 expressing) Th17 cells.     

Physiological and performance changes from the addition of a sprint interval program to wrestling training

Increasing the level of physical fitness for competition is the primary goal of any conditioning program for wrestlers. Wrestlers often need to peak for competitions several times over an annual training cycle. Additionally, the scheduling of these competitions does not always match an ideal periodization plan and may require a modified training program to achieve a high level of competitive fitness in a short-time frame. The purpose of this study was to examine the effects of 4 weeks of sprint-interval training (SIT) program, on selected aerobic and anaerobic performance indices, and hormonal and hematological adaptations, when added to the traditional Iranian training of wrestlers in their preseason phase. Fifteen trained wrestlers were assigned to either an experimental (EXP) or a control (CON) group. Both groups followed a traditional preparation phase consisting of learning and drilling technique, live wrestling and weight training for 4 weeks. In addition, the EXP group performed a running-based SIT protocol. The SIT consisted of 6 35-m sprints at maximum effort with a 10-second recovery between each sprint. The SIT protocol was performed in 2 sessions per week, for the 4 weeks of the study. Before and after the 4-week training program, pre and posttesting was performed on each subject on the following: a graded exercise test (GXT) to determine VO(2)max, the velocity associated with V(2)max (νVO(2)max), maximal ventilation, and peak oxygen pulse; a time to exhaustion test (T(max)) at their νVO(2)max; and 4 successive Wingate tests with a 4-minute recovery between each trial for the determination of peak and mean power output (PPO, MPO). Resting blood samples were also collected at the beginning of each pre and posttesting period, before and after the 4-week training program. The EXP group showed significant improvements in VO(2)max (+5.4%), peak oxygen pulse (+7.7%) and T(max) (+32.2%) compared with pretesting. The EXP group produced significant increases in PPO and MPO during the Wingate testing compared with pretesting (p < 0.05). After the 4-week training program, total testosterone and the total testosterone/cortisol ratio increased significantly in the EXP group, whereas cortisol tended to decrease (p = 0.06). The current findings indicate that the addition of an SIT program with short recovery can improve both aerobic and anaerobic performances in trained wrestlers during the preseason phase. The hormonal changes seen suggest training-induced anabolic adaptations.     

In vivo virus structures: simultaneous classification, resolution enhancement, and noise reduction in whole-cell electron tomography

Hutchinson-Gilford progeria syndrome (HGPS) is a premature aging syndrome caused by the expression and accumulation of a mutant form of lamin A, Δ50 lamin A. As a component of the cell's nucleoskeleton, lamin A plays an important role in the mechanical stabilization of the nuclear envelope and in other nuclear functions. It is largely unknown how the characteristic 50 amino acid deletion affects the conformation of the mostly intrinsically disordered tail domain of lamin A. Here we perform replica exchange molecular dynamics simulations of the tail domain and determine an ensemble of semi-stable structures. Based on these structures we show that the ZMPSTE 24 cleavage site on the precursor form of the lamin A tail domain orients itself in such a way as to facilitate cleavage during the maturation process. We confirm our simulated structures by comparing the thermodynamic properties of the ensemble structures to in vitro stability measurements. Using this combination of experimental and computational techniques, we compare the size, heterogeneity of size, thermodynamic stability of the Ig-fold, as well as the mechanisms of force-induced denaturation. Our data shows that the Δ50 lamin A tail domain is more compact and displays less heterogeneity than the mature lamin A tail domain. Altogether these results suggest that the altered structure and stability of the tail domain can explain changed protein-protein and protein-DNA interactions and may represent an etiology of the disease. Also, this study provides the first molecular structure(s) of the lamin A tail domain, which is confirmed by thermodynamic tests in experiment.     

Assessment of atherosclerotic plaque burden with an elastin-specific magnetic resonance contrast agent

Atherosclerosis and its consequences remain the main cause of mortality in industrialized and developing nations. Plaque burden and progression have been shown to be independent predictors for future cardiac events by intravascular ultrasound. Routine prospective imaging is hampered by the invasive nature of intravascular ultrasound. A noninvasive technique would therefore be more suitable for screening of atherosclerosis in large populations. Here we introduce an elastin-specific magnetic resonance contrast agent (ESMA) for noninvasive quantification of plaque burden in a mouse model of atherosclerosis. The strong signal provided by ESMA allows for imaging with high spatial resolution, resulting in accurate assessment of plaque burden. Additionally, plaque characterization by quantifying intraplaque elastin content using signal intensity measurements is possible. Changes in elastin content and the high abundance of elastin during plaque development, in combination with the imaging properties of ESMA, provide potential for noninvasive assessment of plaque burden by molecular magnetic resonance imaging (MRI).     

Regional Index of Ecological Integrity: A need for sustainable management of natural resources

An ecosystem is a complex composition of physical, chemical and biological components. This complex system remains in a healthy state if the system can maintain the ecological equilibrium among its components. Anthropogenic disturbances are the prime stressors that affect this equilibrium through creating fragmentation, ecosystem sensitivity, loosening landscape connectivity and disrupting ecological integrity. As different types of ecosystem are interconnected, a comprehensive monitoring and evaluating criteria is needed for measuring its integrity at regional level for conservation planning. A Regional Index of Ecological Integrity can be a suitable approach for sustainable management of regional ecosystem. Therefore, this paper presents (i) the characteristics of ecological integrity, (ii) the spatial processes induced by anthropogenic stressors and (iii) an approach to develop a composite Regional Index of Ecological Integrity (RIEI). The prime objective is to establish a thought and a way to develop a composite index of ecological integrity at the regional level. Here, we demonstrate different compositional, structural and functional indicators/indices related to fragmentation, representativeness of protected area, ecosystem sensitivity, and landscape connectivity for the development of a Regional Index of Ecological Integrity (RIEI).