Localized Surface Plasmon Resonance (LSPR)-Based Nanobiosensor for Methamphetamin Measurement

Aptamers are DNA or RNA single-stranded molecules that bind specifically to target molecules with high affinity. Function of nucleic acid aptamers is based on organized tertiary structure of them that is related to primary sequence, length of nucleic acid molecule, and environmental conditions. Herein, a localized surface plasmon resonance (LSPR) nanobioprobe has been developed based on specific aptamer-conjugated gold nanoparticles for rapid detection of methamphetamine. Detection of methamphetamine was studied via monitoring the gold nanoparticles (GNPs) LSPR band alterations in the presence of different concentrations. The covalent conjugation has been confirmed with FT-IR spectroscopy, and size alterations of gold nanoparticles before and after the conjugation state were monitored using dynamic light scattering (DLS) technique. The results show high affinity of aptamer to methamphetamine. Moreover, the results show conjugated aptamer with GNP in different concentrations of methamphetamine that contribute to color changes that is visible with unaided eye. Also, 14 nm LSPR shift was seen after conjugation of aptamer with GNP. Nanoparticle diameter after conjugation with aptamer was increased from 30 to 91 nm and decreased after incubation with methamphetamine (due to folding) from 91 to 84 nm. Detection limit of this designed nanoprobe is 500 nM. Plasmonic nanoparticle-based nanobioprobe is a new field for development of sensitive detection systems.

     

Genetic Background of Hirschsprung Disease: A Bridge Between Basic Science and Clinical Application

Hirschsprung's disease (HSCR) is a congenital disorder, defined by partial or complete loss of the neuronal ganglion cells in the intestinal tract, which is caused by the failure of neural crest cells to migrate completely during intestinal development during fetal life. HSCR has a multifactorial etiology, and genetic factors play a key role in its pathogenesis; these include mutations within several gene loci. These have been identified by screening candidate genes, or by conducting genome wide association (GWAS) studies. However, only a small portion of them have been proposed as major genetic risk factors for the HSCR. In this review, we focus on those genes that have been identified as either low penetrant or high penetrant variants that determine the risk of Hirschsprung's disease. J. Cell. Biochem. 119: 28–33, 2018. © 2017 Wiley Periodicals, Inc.     

<Emphasis Type="Italic">In vitro</Emphasis> study of HAX1 gene therapy by retro viral transduction as a therapeutic target in severe congenital neutropenia

Severe congenital neutropenia (SCN) is a primary immunodeficiency disease in which a number of underlying gene defects are responsible for abnormalities in neutrophil development. The HCLS1-associated protein X1 (HAX1) mutation is associated with an autosomal-recessive form of SCN. Considering the potential of gene therapy approaches for the treatment of monogenic disorders, in this study we aimed to develop retroviral vectors expressing coding sequences (CDS) to be used for the removal of the genetic blockade in deficient hematopoietic cells. Following amplification of CDS with primers containing appropriate restriction sites, HAX1 CDS was cloned into an intermediate vector using TA-cloning. The sequence was transferred into a retroviral vector, followed by retroviral packaging in Plat-A cells. To show HAX1 protein expression, HEK293T cells were exposed to 10 multiplicity of infection (MOI) of retroviral particles and HAX1 expression was confirmed in these cells, using indirect intracellular flow cytometry. This vector was applied for in vitro transduction of hematopoietic stem cell with HAX1 mutation; after 11 days, cultured cells were analyzed for CD66acde and CD177 (neutrophil surface markers) expression. Increased neutrophil production in HAX1 viral vector-expressing hematopoietic cells was observed as compared to control vector transduced cells. Hence, according to the results, this type of therapy could be considered a potential treatment protocol for the disease.     

Bacterial components as naturally inspired nano-carriers for drug/gene delivery and immunization: Set the bugs to work?

Drug delivery is a rapidly growing area of research motivated by the nanotechnology revolution, the ideal of personalized medicine, and the desire to reduce the side effects of toxic anti-cancer drugs. Amongst a bewildering array of different nanostructures and nanocarriers, those examples that are fundamentally bio-inspired and derived from natural sources are particularly preferred. Delivery of vaccines is also an active area of research in this field. Bacterial cells and their components that have been used for drug delivery, include the crystalline cell-surface layer known as “S-layer”, bacterial ghosts, bacterial outer membrane vesicles, and bacterial products or derivatives (e.g. spores, polymers, and magnetic nanoparticles). Considering the origin of these components from potentially pathogenic microorganisms, it is not surprising that they have been applied for vaccines and immunization. The present review critically summarizes their applications focusing on their advantages for delivery of drugs, genes, and vaccines.     

THY1 as a reliable marker for enrichment of undifferentiated spermatogonia in the goat

Spermatogonial stem cells are unique cells of testes that can restore fertility upon transplantation into recipient testes. However, use of suitable markers for enrichment of these cells have important potential application. THY1, is an established conserved marker of spermatogonial stem cells in bovine, rodents, and primates, but there is no information available in goats. After three rounds of enzymatic digestion of prepubertal goat testicular tissues, undifferentiated spermatogonia positive for THY1 were isolated by magnetic-activated cell sorting and were used for immunocytochemistry, real-time polymerase chain reaction analysis for gene expression, protein expression, and transplantation into recipient mice. Immunocytochemical analyses showed that significantly higher percentage of THY1⁺ cells were positive for PLZF and VASA when compared with unselected population. This result for PLZF was further confirmed at the protein level. Real-time polymerase chain reaction analysis revealed that expression of THY1, PLZF, VASA, BCL6B, and UCHL1 as SCCs characteristic genes in THY1⁺ cells was significantly higher than in the initial population. Finally, transplantation of PKH26-labeled cells revealed that THY1⁺ cells had higher capacity for colony formation when compared with unselected cells. In conclusion, the results provide indications that THY1 surface marker can be reliably used for enrichment of undifferentiated spermatogonial in the goats.     

Reaction and survival of four types of sunflowers against Sclerotinia sclerotiorum under controlled conditions

Sclerotinia wilt of sunflower caused by Sclerotinia sclerotiorum (Lib.) De Bary is the major disease of sunflower (Helianthus annuus L.) in Iran. North-western areas of Iran demonstrate the most yield losses through the disease. Development of cultivars with adequate genetic resistance is necessary to avoid these losses. Evaluation of sunflower genotypes varies based on researchers and environmental conditions. Meanwhile, greenhouse tests of the sunflower genotypes are more reliable because of controlled conditions during the assessment activities. To study the reaction of the host plants under controlled conditions, three stem inoculation techniques, including mycelium plug (MP), oxalic acid solution (OAS) and wheat seeds infested (IWS) with Sclerotinia mycelium, were employed. Wounded and non-wounded treatments were used in the experiment to find their effect on the disease progress. Four genotypes, including Ghalami (local variety in market), Confeta, Alstar and Master, were inoculated in this study. The factor was the lesion length to evaluate the effectiveness of different inoculation procedures. The lesion length was measured after 3, 7, 10 and 14 days post inoculation. The analysis of variance demonstrated significant differences between IWS and two other methods (MP and OAS), where the IWS produced the longest lesion lengths. In contrast, the non-significant differences between MP and OAS methods might help researchers to employ the pathogenicity factor (oxalic acid) as an alternative inoculum for their studies. Master variety was the most tolerant genotype among the treatments and its viability was 100% even at 14?days after inoculation and incubation. Interestingly, there was no mortality in all cultivars before 7-day incubations, regardless of resistant or susceptible reactions.     

A web-based identification programme for Pythium species

The genus Pythium, with slightly over 140 described species, has been classified traditionally with other filamentous, coenocytic, sporangia-producing fungi as “Phycomycetes”. However, with recent advances in chemical, ultrastructural and molecular studies, Pythium spp. are now considered as “fungus-like organisms” or “pseudo-fungi” and are placed in the kingdom Chromista or kingdom Stramenopila, distinct from the true fungi or the kingdom Fungi. There is no comprehensive web-based identification key available for the identification of all described Pythium species. This article describes a web-based identification tool (which has been described by the authors under the name of Fungid) that uses morphological features. For the isolate in question, the software compares the unknown isolate with near to 90 known species submitted in the database. One or more than one species is suggested to the user by the software.