FIBS-enabled Noninvasive Metabolic Profiling

In the era of computational biology, new high throughput experimental systems are necessary in order to populate and refine models so that they can be validated for predictive purposes. Ideally such systems would be low volume, which precludes sampling and destructive analyses when time course data are to be obtained. What is needed is an in situ monitoring tool which can report the necessary information in real-time and noninvasively. An interesting option is the use of fluorescent, protein-based in vivo biological sensors as reporters of intracellular concentrations. One particular class of in vivo biosensors that has found applications in metabolite quantification is based on Förster Resonance Energy Transfer (FRET) between two fluorescent proteins connected by a ligand binding domain. FRET integrated biological sensors (FIBS) are constitutively produced within the cell line, they have fast response times and their spectral characteristics change based on the concentration of metabolite within the cell. In this paper, the method for constructing Chinese hamster ovary (CHO) cell lines that constitutively express a FIBS for glucose and glutamine and calibrating the FIBS in vivo in batch cell culture in order to enable future quantification of intracellular metabolite concentration is described. Data from fed-batch CHO cell cultures demonstrates that the FIBS was able in each case to detect the resulting change in the intracellular concentration. Using the fluorescent signal from the FIBS and the previously constructed calibration curve, the intracellular concentration was accurately determined as confirmed by an independent enzymatic assay.     

Heat stress aggravates oxidative stress,apoptosis, and endoplasmic reticulum stress in the cerebellum of male C57 mice

Molecular Biology Reports - The current study was set to assess the effect of heat stress exposure on oxidative stress, apoptosis, and endoplasmic reticulum stress markers in the cerebellum of male...     

Comparison of PCR-RFLP with 21-plex PCR and rDNA Sequencing for Identification of Clinical Yeast Isolates

Non-albicans Candida species and other rare yeasts have emerged as major opportunistic pathogens in fungal infections. Identification of opportunistic yeasts in developing countries is mainly performed by phenotypic assay, which are time-consuming and prone to errors. The aim of the present study was to evaluate PCR-RFLP as a routinely used identification technique for the most clinically important Candida species in Iran and make a comparison with a novel multiplex PCR, called 21-plex PCR. One hundred and seventy-three yeast isolates from clinical sources were selected and identified with sequence analysis of the D1/D2 domains of rDNA (LSU rDNA) sequencing as the gold standard method. The results were compared with those obtained by PCR-RFLP using MspI restriction enzyme and the 21-plex PCR. PCR-RFLP correctly identified 93.4% of common pathogenic Candida species (C. albicans, C. glabrata, C. parapsilosis, C. tropicalis, and P. kudriavsevii (=?C. krusei)) and was able to identify 45.5% of isolates of the uncommon yeast species compared to the D1/D2 rDNA sequencing. Compared with PCR-RFLP, all common Candida species and 72.7% of uncommon yeast species were correctly identified by the 21-plex PCR. The application of the 21-plex PCR assay as a non-sequence-based molecular method for the identification of common and rare yeasts can reduce turnaround time and costs for the identification of clinically important yeasts and can be applied in resource-limited settings.

     

The Arabidopsis expansin gene (AtEXPA18) is capable to ameliorate drought stress tolerance in transgenic tobacco plants

Molecular Biology Reports - Expansins are cell wall proteins loosening plant cell in pH-dependent manner. This study aimed to investigate the role of AtEXPA18 in different morphological,...     

Biomechanical evaluation of porous biodegradable scaffolds for revision knee arthroplasty

Tibial bone defect is a critical problem for revision knee arthroplasty. Instead of using metallic spacer or cement, biodegradable scaffolds could be an alternative solution. A numerical model of a revision knee arthroplasty was thus developed to estimate the mechanical resistance of the scaffold in this demanding situation. The tibia, scaffold, and prosthesis were represented by simplified parameterised geometries. The maximal gait cycle force was applied asymmetrically to simulate a critical loading. Several parameters were analysed: 1) inter-individual variability, 2) cortical bone stiffness, 3) cortical bone thickness, 4) prosthesis fixation quality, and 5) scaffold thickness. The calculated scaffold strain was compared to its experimental ultimate strain. Among the tested parameters, failure was only predicted with scaffold thickness below 5 mm. This study suggests that biodegradable bone scaffolds could be used to fill bone defects in revision knee arthroplasty, but scaffold size seems to be the limiting factor.     

Fatal Invasive Pulmonary Aspergillosis in COVID-19 Patient with Acute Myeloid Leukemia in Iran

Mycopathologia - Although patients with severe immunodeficiency and hematological malignancies has been considered at highest risk for invasive fungal infection, patients with severe pneumonia due...     

Screening of oilseed rape cultivars relation to diamondback moth,Plutella xylostella (Lep.: Plutellidae) in fields of Tehran

The diamondback moth (DBM), Plutella xylostella (L.) (Lep., Plutellidae), is one of the most important pests of oilseed rape, Brassica napus L., in the world. In this study, resistance of oilseed rape cultivars to DBM was evaluated in fields of Tehran based on injury rate of the cultivars. To evaluate cultivar resistance, 19 cultivars of oilseed rape were planted in randomised complete-block design with three replications. Total experiments were conducted in experimental field of Shahed University. Results showed that DBM had activity on the plants during season and peak of the injury occurred on 27 May. Results indicated that the total of oilseed rape cultivar was spotty. Statistical analysis showed that there was significant difference among determined characteristics in all cultivars. The highest and lowest numbers of holes in the leaf were observed in Midas, Karaj3, Zarfam and Hyola420, Hyola308 cultivars, respectively, in sampling of the end growth. Opera and Midas cultivars have the highest percentage of leaf infestation and the lowest percentage of leaf infestation was observed in Hyola420 and Hyola308. Also, Karaj3 and Zarfam cultivars have the highest percentage of meristem infestation and the lowest percentage of meristem infestation was observed in Licard and Hyola420 cultivars. Based on peak of the injury rate and importance of meristem injury, Hyola420 and Hyola308 cultivars were relatively resistant to DBM, and Karaj3 and Zarfam were relatively sensitive to DBM.     

Understory vegetation as an indicator of soil characteristics in the Hyrcanian area,N. Iran

The mesic Caspian (Hyrcanian) forest and ecotone communities provide a marked contrast to the arid and semiarid landscapes associated with most of the territory of Iran. To date, the ecological characteristics of these habitats, threatened and of conservation importance, have been little studied. Accordingly, ecological profiles of some important plant species of these communities have been assessed along two altitudinal gradients (300–2300 m a.s.l.). Vegetation and soils were sampled every 100 m in elevation, with the data subsequently analyzed using TWINSPAN and corrected frequency (CF) analyses. Relationships between soil variables (subdivided into three classes, the lowest, the middle and the upper third of all values) and herbaceous and shrub species (presence/absence data) were analyzed by the polythetic divisive classification method. 379 plant species and eleven soil variables – N, P, K, CaCO₃, EC, pH, organic matter, C/N ratio and percentage of sand, clay and silt – were considered. The ecological profile method was used to evaluate the affinity and significance of associations between the probability of species’ occurrence and topsoil characteristics found by the polythetic method. Five vegetation groups were identified: two groups, with Acer campestre and Quercus macranthera in the tree layer and Veronica mazanderanae and Phuopsis stylosa as herbs, were restricted to forest-steppe ecotones and the upper mountain areas. Three groups, with Acer velutinum, Ruscus hyrcanus, Carpinus betulus, Danae racemosa, Fagus orientalis and Aruncus vulgaris as indicator species, occurred in the forest itself. Of the 42 plant species assessed as being of particular importance, 13 had significant relationships with eight soil factors. Thus, certain species, including endemic plant species of restricted distribution and conservation importance, can be used as indicators of particular soil conditions in the Hyrcanian forest area.     

Olfactory response of Phytoseiulus persimilis (Acari: Phytoseiidae) to untreated and Beauveria bassiana-treated Tetranychus urticae (Acari: Tetranychidae)

Determination of attraction and avoidance behavior of predators is important in concomitant use of multiple natural enemies to control a pest. The olfactory response of the predatory mite Phytoseiulus persimilis was studied to odors related to Tetranychus urticae adults infected by Beauveria bassiana DEBI008 in 0, 24, 48 and 72 h intervals, both in absence and in presence of plants. In plant-present experiments, P. persimilis attraction was neither towards adults of T. urticae infected by 0.02 % Tween 80 (as control), nor to the ones infected by B. bassiana for 0 or 24 h, whereas significant attraction towards the control was observed when tested against T. urticae infected by B. bassiana for 48 or 72 h. In absence of plants, P. persimilis displayed significant avoidance of T. urticae infected by B. bassiana for 48 or 72 h, when their alternative option was 0.02 % Tween 80-infected T. urticae adults. These results indicate that P. persimilis can recognize the presence of B. bassiana and that the predator avoids the fungus. This suggests that the two natural enemy species can be used together in biological control programmes.