Optimizing methods for human testicular tissue cryopreservation and spermatogonial stem cell isolation

Cryopreservation of testicular tissue before cancer therapy for fertility preservation in prepubertal boys with cancer is of great interest in reproductive medicine. Isolation of spermatogonial stem cells (SSCs) from cryopreserved tissues would be a suitable cell source to re-establish spermatogenesis after cancer therapy. We herein establish optimized protocols for cryopreservation of human testicular tissue and isolation of SSCs from cryopreserved tissue. We developed a freezing protocol that provided high testicular cell viability and supported structural integrity and tubular epithelium coherence similar to fresh tissue. Then, we established a protocol that allowed efficient isolation of functional SSCs from cryopreserved tissues. Isolated cells were found on the testicular basement membrane after xenotransplantation. Our results demonstrated the preservation of testicular tissue structure and high cell viability with efficient isolation of SSCs after testicular cryopreservation, which is promising for future therapeutic applications in fertility preservation.     

Survey of cytotoxic and UV protection effects of biosynthesized cerium oxide nanoparticles

Cerium oxide nanoparticles (CeO₂ NPs) are among the important nanoparticles that are extensively utilized in cosmetics, automotive industries, ultraviolet (UV) filtration, gas sensors, and pharmaceutical products. In this study, CeO₂ NPs were synthesized using an aqueous extract of Ziziphus jujube fruit. The synthesized nanoparticles were characterized using UV‐visible spectroscopy, powder X‐ray diffraction, Fourier transform infrared spectroscopy, energy‐dispersive spectroscopy, field energy scanning electron microscopy, and Raman methods. The results indicated that the size of synthesized nanoparticles is between 18 and 25 nm, and they have a spherical shape. UV absorbance of the synthesized nanoparticles was measured through spectrophotometric method in the range of 290 to 320 nm. The cytotoxic activity of synthesized CeO₂ NPs against colon (HT‐29) cancer cell line was surveyed through 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide assay. The results showed that synthesized nanoparticles are nontoxic on HT‐29 cells under 400 μg/mL concentrations after 24 hours of treatment time periods. The increase in treatment time cases increases cytotoxic activity of synthesized nanoparticles. Sun protection factor of CeO₂ NPs, as a criterion for amount of sunlight radiation protection, was determined by applying Mansur equation. The results demonstrated that synthesized CeO₂ NPs have excellent UV protection and sunscreen physical absorption properties.     

Effect of Encapsulated Lactobacillus bulgaricus on Innate Immune System and Hematological Parameters in Rainbow Trout (Oncorhynchus mykiss), Post-Administration of Pb

Probiotics and Antimicrobial Proteins - The present study was conducted to investigate the effects of probiotic and encapsulated Lactobacillus bulgaricus on hematological and immunological factors...     

Novel delivery systems for interferons

Interferons, IFNs, are among the most widely studied and clinically used biopharmaceuticals. Despite their invaluable therapeutic roles, the widespread use of IFNs suffers from some inherent limitations, mainly their relatively short circulation lifespan and their unwanted effects on some non-target tissues. Therefore, both these constraints have become the central focus points for the research efforts on the development of a variety of novel delivery systems for these therapeutic agents with the ultimate goal of improving their therapeutic end-points. Generally, the delivery systems currently under investigation for IFNs can be classified as particulate delivery systems, including micro- and nano-particles, liposomes, minipellets, cellular carriers, and non-particulate delivery systems, including PEGylated IFNs, other chemically conjugated IFNs, immunoconjugated IFNs, and genetically conjugated IFNs. All these strategies and techniques have their own possibilities and limitations, which should be taken into account when considering their clinical application. In this article, currently studied delivery systems/techniques for IFN delivery have been reviewed extensively, with the main focus on the pharmacokinetic consequences of each procedure.     

Oligo-Miocene fluvial-dominated deltas on the shelf of the South Caspian Sea (Paratethys)

The Oligo-Miocene Zivah Formation forms the principal reservoir unit in the Moghan area of northwest Iran. Characterization of the constituent facies of the formation and their spatial distribution represent a major aspect of this investigation. The combination of results from the field and laboratory studies lead to the determination of 18 lithofacies and 13 petrofacies (petrographic lithofacies). Three major types of sedimentary cycles (small-, medium-, and large-scale cycles) of variable thickness (few meters, few tens of meters, few hundreds of meters, respectively) are recognized on the basis of facies association properties. Depositional environments representing each facies and facies association are defined and interpreted on the basis of their sedimentological characteristics. The major depositional systems of the Zivah Formation are interpreted as fluvial-dominated deltas on the shelf of the South Caspian Sea (or Paratethys). The principal controls on sedimentation are discussed in terms of lithofacies/petrofacies characteristics and the nature of sedimentary cycles. Development of large-scale cycles is related to world-wide sea-level change (eustasy), medium-scale cycles to tectonism along the basin margin, and small-scale cycles to the rate of sediment supply and local environmental conditions. Active tectonism seems to be the main cause for high sediment supply to the Zivah Basin.     

A novel spectrophotometric method for determination of kinetic constants of aldehyde oxidase using multivariate calibration method

Although phenanthridine has been frequently used as a specific substrate for the assessment of aldehyde oxidase activity, the use of this method is questionable due to a lower limit of detection and its validity for kinetic studies. In the present study, a novel sensitive multivariate calibration method based on partial least squares (PLS) has been developed for the measurement of aldehyde oxidase activity using phenanthridine as a substrate. Phenanthridine and phenanthridinone binary mixtures were prepared in a dynamic linear range of 0.1–30.0 μM and the absorption spectra of the solutions were recorded in the range of 210–280 nm in Sorenson's phosphate buffer (pH 7.0) containing EDTA (0.1 mM). The optimized PLS calibration model was used to calculate the concentration of each chemical in the prediction set. Hepatic rat aldehyde oxidase was partially purified and the initial oxidation rates of different concentrations of phenanthridine were calculated using the PLS method. The values were used for calculating Michaelis–Menten constants from a Lineweaver–Burk double reciprocal plot of initial velocity against the substrate concentration. The limits of detection for phenanthridine and phenanthridinone were found to be 0.04 ± 0.01 and 0.03 ± 0.01 μM (mean ± SD, n = 5), respectively. Using this method, the K_m value for the oxidation of phenanthridine was calculated as 1.72 ± 0.09 μM (mean ± SD, n = 3). Thus, this study describes a novel spectrophotometric method that provides a suitable, sensitive and easily applicable means of measuring the kinetics of phenanthridine oxidation by aldehyde oxidase without the need for expensive instrumentation.     

State of the art technologies to explore long non‐coding RNAs in cancer

Long non‐coding RNAs (lncRNAs) comprise a vast repertoire of RNAs playing a wide variety of crucial roles in tissue physiology in a cell‐specific manner. Despite being engaged in myriads of regulatory mechanisms, many lncRNAs have still remained to be assigned any functions. A constellation of experimental techniques including single‐molecule RNA in situ hybridization (sm‐RNA FISH), cross‐linking and immunoprecipitation (CLIP), RNA interference (RNAi), Clustered regularly interspaced short palindromic repeats (CRISPR) and so forth has been employed to shed light on lncRNA cellular localization, structure, interaction networks and functions. Here, we review these and other experimental approaches in common use for identification and characterization of lncRNAs, particularly those involved in different types of cancer, with focus on merits and demerits of each technique.     

Establishment and in vitro differentiation of a new embryonic stem cell line from human blastocyst

Embryonic stem cells have the ability to remain undifferentiated and proliferate indefinitely in vitro while maintaining the potential to differentiate into derivatives of all three embryonic germ layers. These cells have, therefore, potential for in vitro differentiation studies, gene function, and so on. The aim of this study was to produce a human embryonic stem cell line. An inner cell mass of a human blastocyst was separated and cultured on mouse embryonic fibroblasts in embryonic stem cell medium with related additives. The established line was evaluated by morphology; passaging; freezing and thawing; alkaline phosphatase; Oct-4 expression; anti-surface markers including Tra-1-60 and Tra-1-81; and karyotype and spontaneous differentiation. Differentiated cardiomyocytes and neurons were evaluated by transmission electron microscopy and immunocytochemistry. Here, we report the derivation of a new embryonic stem cell line (Royan H1) from a human blastocyst that remains undifferentiated in morphology during continuous passaging for more than 30 passages, maintains a normal XX karyotype, is viable after freezing and thawing, and expresses alkaline phosphatase, Oct-4, Tra-1-60, and Tra-1-81. These cells remain undifferentiated when grown on mouse embryonic fibroblast feeder layers in the presence or absence of recombinant human leukemia inhibitory factor. Royan H1 cells can differentiate in vitro in the absence of feeder cells and can produce embryoid bodies that can further differentiate into beating cardiomyocytes as well as neurons. These results define Royan H1 cells as a new human embryonic stem cell line.     

A hardwickiic acid derivative from Pulicaria gnaphalodes

The aerial parts of Pulicaria gnaphalodes afrorded a new clerodane lactone closely related to hardwickiic acid.