Alteration in genes expression patterns during in vitro differentiation of mouse spermatogonial cells into neuroepithelial-like cells

Pluripotent stem cells derived from testis is a new, natural, and unlimited source for cell therapy in regenerative medicine and represent a possible alternative to replacing of all cells in the body. Here, we designed a simple co-culture system of spermatogonia cells with Sertoli cells for the generation of embryonic stem-like cells from mouse testis. The importance of our simple method will be clear when we compared it with other complex and time-consuming methods. Embryonic stem-like colonies with sharp border confirmed by real-time PCR, immunocytochemistry and flow cytometry assessments. Embryonic stem-like colonies were immunopositive for pluripotency markers. Transition of spermatogonia cells to embryonic stem-like cells was accompanied by extensive changes in gene expression. These changes included significant increase in pluripotency genes expression and significant decrease in germ cell-specific genes expression. Also, we proved the differentiation capacity of embryonic stem-like cells to neuroepithelial-like cells which were immunoreactive to Nestin and Neurofilament 68. Evaluation of genes expression during in vitro differentiation into neuroepithelial-like cells showed high-level expression of Nestin whether this gene approximately has no expression in undifferentiated embryonic stem-like cells. Also, expression of pluripotency genes has significantly decreased in neuroepithelial-like cells compared with embryonic stem-like cells. This study shows that embryonic stem-like cells derived from testis are capable to differentiate into neuroepithelial-like cells that may provide a cellular reservoir usable for neurodegenerative disorders.     

A genetic assay of three patients in the same family with Holt-Oram syndrome; a case report

Holt-Oram syndrome (HOS) is a developmental disorder inherited in an autosomal-dominant pattern. Affected organs are the heart and forelimbs with upper extremity skeletal defects and congenital heart malformation. In this study we present three cases of HOS in the same family. In one of these three individuals we detected a transition of C to T (CTG-GTT, V205V) in exon 7 of the TBX5 gene. This nucleotide change causes no amino acid change and potential pathologic effects remain unknown.Key Words: Holt-Oram syndrome, Congenital heart malformation, TBX5 gene     

Effects of the adsorption of alkali metal oxides on the electronic,optical, and thermodynamic properties of the Mg<Subscript>12</Subscript>O<Subscript>12</Subscript>nanocage: a density functional theory study

The effect of alkali metal oxides M _n O (M?=?Li, Na, K; n?=?2, 3, 4) on the geometric, electronic, and linear and nonlinear optical properties of the Mg₁₂O₁₂ nanocage was investigated by density-functional-based methods. According to the computational results, these alkali metal oxides are adsorbed on the Mg₁₂O₁₂ nanocage because this adsorption reduces its energy gap. The static first hyperpolarizability (β ₀) of the nanocage is dramatically increased in the presence of the alkali metal oxides, with the greatest increase seen in the presence of the superalkalis (i.e., M₃O; M?=?Li, Na, and K). The highest first hyperpolarizability (β ₀?≈?600,000 a.u.) was calculated for K₃O@Mg₁₂O₁₂, which was considerably more than that for Mg₁₂O₁₂. The thermodynamic properties and relative stabilities of these inorganic compounds are discussed.

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Graphical Abstract Optimized structure and DOS spectrum of K₃O(e@Mg₁₂O₁₂)

     

Measurement of some Benzylisoquinoline Alkaloids in Different Organs of Persian Poppy during Ontogenetical Stages

Papaver bracteatum, a perennial species, has been known as a rich source of thebaine and a potential alternative to Papaver somniferum for the production of codeine and some semisynthetic antagonist drugs. In this study, ion mobility spectrum (IMS) of the root, leaf, bottom part of stem, upper part of stem, capsule wall, petal, and capsule content during developmental stages of P. bracteatum including annual rosette, perennial rosette, bud initiation, pendulous bud, preflowering, and lancing were investigated. The IMS revealed thebaine, papaverine, and noscapine as the major components of the extracted alkaloids. Based on the results of the study it appears that, at least in part, there is a competition among the biosynthesis pathways of papaverine, noscapine, and morphinan alkaloids from a common source . Root and capsule wall were the most potent organs for extraction of thebaine, while lancing stage was the best developmental stage for thebaine exploitation. However, it seems that total biomass of root and capsule wall plays a key role in the final selection of favorite organ. Although papaverine and noscapine in the stem at preflowering stage had the most quantity, significant amounts were found in the capsule wall. In general, total alkaloid content of leaf was lower than the other plant parts.     

Variation in Essential Oil Yield and Composition of Dorema aucheri Boiss., an Endemic Medicinal Plant Collected from Wild Populations in Natural Habitats

In the present research, variability in essential oil (EO) composition of five Dorema aucheri populations collected from natural habitats in different regions of Iran, were investigated. The EO content of populations varied from 0.28 to 0.68%. According to gas chromatography/mass spectrometry analysis, β‐caryophyllene (7.17 – 35.73%), thymol (23.45 – 29.64%), β‐gurjunene (2.58 – 5.89%), carvacrol (1.32 – 2.67%) and cuparene (1.97 – 2.98%) were the major components. Hierarchical cluster, principal component and canonical correspondence analyses classified the studied populations into three groups based on major EO components. The environmental parameters of the collected sites were also evaluated. According to the results, it might be suggested that sandy soils with high mean annual precipitation were major environmental factors influencing the amount of β‐caryophyllene, while thymol, cuparene and caryophyllen oxide increased in silty and clay soils. Finally, the population collected in high altitudes and clay soils had higher amount of β‐gurjunene.     

Cobalt separation by <Emphasis Type="Italic">Alphaproteobacterium</Emphasis> MTB-KTN90: magnetotactic bacteria in bioremediation

Bioremediation of toxic metals by magnetotactic bacteria and magnetic separation of metal-loaded magnetotactic bacteria are of great interest. This bioprocess technique is rapid, efficient, economical, and environmentally friendly. In this study, cobalt removal potential of a novel isolated magnetotactic bacterium (Alphaproteobacterium MTB-KTN90) as a new biosorbent was investigated. The effects of various environmental parameters in the cobalt removal and the technique of magnetic separation of cobalt-loaded bacterial cells were studied. Cobalt removal by MTB-KTN90 was very sensitive to pH solution; higher biosorption capacity was observed around pH 6.5–7.0. When biomass concentration increased from 0.009 to 0.09 g/l, the biosorption efficiency increased from 13.87 % to 19.22 %. The sorption of cobalt by MTB-KTN90 was rapid during the first 15 min (859.17 mg/g dry weight). With the increasing of cobalt concentrations from 1 to 225 mg/l, the specific cobalt uptake increased. Maximum cobalt removal (1160.51 ± 15.42 mg/g dry weight) took place at optimum conditions; pH 7.0 with initial cobalt concentration of 115 mg/l at 60 min by 0.015 g/l of dry biomass. The results showed maximum values for constants of Langmuir and Freundlich models so far. The biosorption mechanisms were studied with FTIR, PIXE, and FESEM analysis. Cobalt-loaded MTB-KTN90 had ability to separate from solution by a simple magnetic separator. Magnetic response in MTB-KTN90 is due to the presence of unique intracellular magnetic nanoparticles (magnetosomes). The orientation magnetic separation results indicated that 88.55 % of cobalt was removed from solution. Consequently, Alphaproteobacterium MTB-KTN90 as a new biosorbent opens up good opportunities for the magnetic removal of cobalt from the polluted aquatic environments.     

Mathematical modeling of cell growth in a 3D scaffold and validation of static and dynamic cultures

Tissue engineering, an immensely important field in contemporary clinical practices, aims at the repair or replacement of damaged tissues. The mathematical model proposed herein shows the distribution and growth of cells in their characteristic time in a 3D scaffold model. This study contributes to the progress of simulation techniques in static and dynamic cultures of bone tissue. Brinkman, nutrient transport, and cell growth equations are brought together to quantify the growth behavior of cells. However, when a static culture is being studied, the Brinkman equation is eliminated. The model was validated by experimental cell culture using 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide assay and scanning electron microscopy. Then, static and dynamic cultures were compared to assess the cell density and cell distribution in the scaffold. Cell counting after 21 days of cell culture showed that the number of cells increased 42‐fold in static and 53.5‐fold in dynamic cultures, which was in good agreement with our model estimations (37‐fold increase in the number of cells in static and 49‐fold increase in dynamic cultures). In conclusion, our mathematical model could predict cell distribution and growth in the scaffold.     

Exonic Splicing Mutations Are More Prevalent than Currently Estimated and Can Be Predicted by Using In Silico Tools

The identification of a causal mutation is essential for molecular diagnosis and clinical management of many genetic disorders. However, even if next-generation exome sequencing has greatly improved the detection of nucleotide changes, the biological interpretation of most exonic variants remains challenging. Moreover, particular attention is typically given to protein-coding changes often neglecting the potential impact of exonic variants on RNA splicing. Here, we used the exon 10 of MLH1, a gene implicated in hereditary cancer, as a model system to assess the prevalence of RNA splicing mutations among all single-nucleotide variants identified in a given exon. We performed comprehensive minigene assays and analyzed patient’s RNA when available. Our study revealed a staggering number of splicing mutations in MLH1 exon 10 (77% of the 22 analyzed variants), including mutations directly affecting splice sites and, particularly, mutations altering potential splicing regulatory elements (ESRs). We then used this thoroughly characterized dataset, together with experimental data derived from previous studies on BRCA1, BRCA2, CFTR and NF1, to evaluate the predictive power of 3 in silico approaches recently described as promising tools for pinpointing ESR-mutations. Our results indicate that ΔtESRseq and ΔHZ_EI-based approaches not only discriminate which variants affect splicing, but also predict the direction and severity of the induced splicing defects. In contrast, the ΔΨ-based approach did not show a compelling predictive power. Our data indicates that exonic splicing mutations are more prevalent than currently appreciated and that they can now be predicted by using bioinformatics methods. These findings have implications for all genetically-caused diseases.