The potential of phytoremediation using hyperaccumulator plants: a case study at a lead-zinc mine site

Contamination with heavy metals is one of the most pressing threats to water and soil resources, as well as human health. Phytoremediation might potentially be used to remediate metal-contaminated sites. A major advance in the development of phytoremediation for heavy metal affected soils was the discovery of heavy metal hyperaccumulation in plants. This study applied several established criteria to identify hyperaccumulator plants. A case study was conducted at a mining area in the Hamedan province in the west central region of Iran. The results indicated that plant metal accumulation differed among species and plant parts. Plant species grown in substrata with elevated metal levels contained significantly higher metal levels. Using the most common criteria, Euphorbia macroclada and Centaurea virgata can be classified as hyperaccumulators of specific heavy metals measured in this study and they might potentially be used for the phytoremediation of contaminated soils.     

Matrix remodeling in experimental and human heart failure: a possible regulatory role for TIMP-3

In the failing heart, an imbalance in matrix metalloproteinases (MMPs) and their biological regulators, the tissue inhibitors of MMPs (TIMPs), may result in cardiac dilatation from matrix degradation. We hypothesized that a reduction of myocardial TIMP-3 is associated with adverse matrix remodeling in both human and experimental heart failure. Cardiomyopathic hamsters at age 15 wk (normal), 25 wk (compensated stage), and 35 wk (overt failure) were compared with age-matched normal controls. MMP activity (gelatinase bioassay) was increased in cardiomyopathic hearts (P = 0.03) and peaked during the transition to overt heart failure. TIMP-3 content (immunoblot) was decreased compared with normal controls (74 +/- 5% at 25 wk, 69 +/- 10% at 35 wk; P = 0.001) and its reduction was associated with increased MMP activity (r = -0.6; P = 0.004). TIMP-1 increased progressively (P = 0.001), whereas TIMP-2, TIMP-4, and MMP protein levels were unchanged. Myocardial collagen (hydroxyproline content) increased with time during the progression to end-stage cardiac failure (P < 0.0001). Collagen synthesis ([(14)C]proline uptake) was elevated in cardiomyopathy at 15 and 25 wk (P < 0.05). The collagen cross-linking ratio (insoluble:soluble collagen) was reduced (P = 0.003) as the left ventricle dilated. By confocal microscopy restricted to viable myocardium, collagen content was reduced (P = 0.04) with fragmentation (P < 0.0001) and thinning (P = 0.003) of perimysial collagen fibers. Similarly, patients with end-stage congestive heart failure (n = 7) compared with nonfailing controls (n = 2) had elevated gelatinase MMP activity (P = 0.02) associated with isolated reductions in TIMP-3 (55 +/- 5% of normal; P = 0.003). Reductions of TIMP-3 parallel adverse matrix remodeling in the cardiomyopathic hamster and the failing human heart. TIMP-3 may contribute to the regulation of myocardial remodeling and its reduction may promote a transition from compensated to end-stage congestive heart failure.     

Generation of tolerogenic dendritic cells using Lactobacillus rhamnosus and Lactobacillus delbrueckii as tolerogenic probiotics

Systemic lupus erythematosus (SLE) concurs with excessive uncontrolled inflammatory immune responses that lead to the loss of immune tolerance. Dendritic cells (DCs) are important and determinant immune cells that regulate immune responses. Tolerogenic DCs with regulatory markers and cytokines could induce regulatory immune cells and responses. Tolerogenic probiotics are capable of producing regulatory DCs from monocytes in in vitro conditions. The purpose of this study was to evaluate the effect of Lactobacillus delbrueckii and Lactobacillus rhamnosus on the production of DCs in an in vitro condition. Peripheral blood mononuclear cells were isolated from the healthy and SLE donors. Monocytes were cultured with optimized concentrations of granulocyte‐macrophage colony‐stimulating factor (GM‐CSF) and interleukin 4 (IL‐4) to produce immature DCs (IDCs). An IDC uptake assay was performed, and IDCs of healthy and SLE donors were divided into three subgroups following 48 hours of treatment with GM‐CSF and IL‐4, along with L. delbrueckii, L. rhamnosus, and mixed probiotics for the production of tolerogenic DCs. The surface expression of Human Leukocyte Antigen‐antigen D Related (HLA‐DR), CD86, CD80, CD83, CD1a, and CD14 was analyzed using flow cytometry, and the gene expression levels of indoleamine 2,3‐dioxygenase (IDO), IL‐10, and IL‐12 were measured using real‐time polymerase chain reaction. We observed significantly reduced expression of costimulatory molecules and other surface markers in the probiotic‐induced mature DCs (MDCs) in both healthy and SLE donor groups in comparison with lipopolysaccharide (LPS)‐induced MDCs. In addition, the expression of IDO and IL‐10 increased, whereas IL‐12 decreased significantly in probiotic‐induced MDCs compared with LPS‐induced MDCs. IDCs and especially mature tolerogenic DC of SLE patients highly expressed IDO. The results of the current study suggested that live probiotics could modify properties of DCs to modulatory cells, which might contribute to the induction of tolerance and renovation of immune hemostasis.     

Differential fluorescence of sister chromatids with 4'-6-diamidino-2-phenylindole.

Differential fluorescence of sister chromatids and sister chromatid exchanges (SCE) in chromosomes from human lymphocytes grown two replication cycles in medium containing 5-bromodeoxyuridine can be detected by fluorescence microscopy after staining with 4'-6-diamidino-2-phenylindole (DAPI). The DAPI fluorescence appears to be more stable than that of the dye 33258 Hoechst and may provide a more sensitive method for the detection of SCE.     

Evidence for genetic control of nondisjunction in man.

Data on factors associated with the occurrence of Down syndrome in a highly inbred population were evaluated to investigate the presence of a genetic control of nondisjunction in man. In Kuwait, close consanguinity occurs in 40% of marriages. In its main obstetric hospital, 20 trisomic Down babies out of 11,614 singleton births were delivered over a 12-month period. Chi-square analyses indicate the occurrence of Down syndrome to be linked to two independent factors: consanquinity of parents and maternal age. The relative risk is approximately four times greater for closely related than for nonrelated parents (P less than .005); a possible explanation for this is the existence of a gene that induces mitotic nondisjunction in the homozygous fertilized ovum. An alternative explanation is the existence of an autosomal recessive gene which results in meiotic nondisjunction in the homozygous parents. Consanguinity is usually perpetuated in certain families, or sections of the population, and parents in highly inbred families have a higher probability to be homozygotes for that gene.     

NMR assignment of iso-alpha-acids from isomerised extracts of Humulus lupulus L. cones

Iso-alpha-acids are known to contribute to the characteristic bitter taste of beer. Six iso-alpha-acids were isolated from a carbon dioxide extract of the cones of Humulus lupulus L. by centrifugal partition chromatography followed by separation through beta-cyclodextrin binding. This method overcame the low yield issue of most isolation procedures that results from the low stability of these compounds to light and oxygen. Their full identification was performed using one- and two-dimensional NMR spectrometry, including (1)H- and (13)C-NMR, (1)H-(1)H COSY, HMQC and HMBC and electrospray ionisation mass spectrometry. The results confirmed the structures of the isolated compounds as trans-isocohumulone, cis-isocohumulone, trans-isohumulone, cis-isohumulone, trans-isoadhumulone and cis-isoadhumulone. Epimers can be easily distinguished by observing the chemical shift differences of the H-5, H-1', H-2' and C-5 signals and the different splitting pattern of H-5 and H-2'.     

Application of BATMAN and BAYESIL for quantitative <Superscript>1</Superscript>H-NMR based metabolomics of urine: discriminant analysis of lean,obese, and obese-diabetic rats

Introduction

BATMAN and BAYESIL are software tools, which can provide a solution for automated metabolite quantifications based on the proton nuclear magnetic resonance (¹H-NMR) spectral data of bio-fluids. However, their specific application for the quantitative ¹H-NMR based metabolomics of urine has not been investigated.

Objectives

The aim of this study is to evaluate the performance of BATMAN and BAYESIL in the quantitative metabolite analysis of urine based on its ¹H-NMR spectra.

Methods

BATMAN and BAYESIL were used for automated metabolite quantification based on the ¹H-NMR spectra of the urine from the lean, obese and obese-diabetic rat groups. PLS-DA model was used to discriminate the three different groups based on the results from the quantifications.

Results

BATMAN was found to be superior to BAYESIL in identifying and quantifying the metabolites in the urine samples, owing to its flexibility that allows users to define and adjust the relevant signals of the pure standard metabolites in the database in order to fit the signals in the samples, a necessary step since variations and peak shift are natural in most ¹H-NMR spectra. The results of BATMAN also agreed well with that of the manual deconvolution method, which indicated the higher accuracy in metabolite quantification, despite the need of pre-processing and longer processing time than BAYESIL. However, in the case where the problems in baseline correction and peak shift of ¹H-NMR spectra are absent, the use of BAYESIL is more advantageous. Application of quantitative ¹H-NMR based metabolomics of the urine showed that PLS-DA model derived from BATMAN could satisfactorily discriminate the lean, obese, and obese-diabetic rat groups.

Conclusion

Both BATMAN and BAYESIL are useful for the quantitative automation of urine metabolites based on its ¹H-NMR spectra. The results from BATMAN method is superior to BAYESIL but require expertise in spectroscopy and longer computer time. Both methods help in simplifying the interpretation of metabolite status in the VIP analysis.

     

Avian embryos and related cell lines: A convenient platform for recombinant proteins and vaccine production

Chick embryos are a significant historical research model in basic and applied sciences. The embryonated eggs have been used for virus inoculation in order to vaccine production for nearly a century. Recently, avian eggs and cell lines derived from embryonated eggs have found wide application in biotechnology. This review will discuss about the unique characteristics of avian eggs in terms of safety, large scale and economical production of recombinant proteins. This system also provides the human‐like glycosylation on target proteins and therefore can be considered as a suitable host for biomanufacturing of humanized monoclonal antibodies and therapeutic proteins. Avian derived cell lines are an alternative for rapid vaccine manufacturing during a pandemic. Based on the latest knowledge in cell and animal transgenesis, the currently available germ cell‐mediated gene transfer system provides a more efficient strategy in gene targeting and creation of transgenic birds that lead to advancements in industrial, biotechnology, and biological research applications. This review covers the recent development of avian fertilized eggs and related cell lines in a variety of human biopharmaceuticals and viral vaccine manufacturing.     

A simple,real-time assay of horseradish peroxidase using biolayer interferometry

ABSTRACT

Horseradish peroxidase (HRP) isoenzyme C1a is one of the most widely used enzymes for various analytical methods in bioscience research and medical fields. In these fields, real-time monitoring of HRP activity is highly desirable because the utility of HRP as a reporter enzyme would be expanded. In this study, we developed a simple assay system enabling real-time monitoring of HRP activity by using biolayer interferometry (BLI). The HRP activity was quantitatively detected on a BLI sensor chip by tracing a binding response of tyramide, a substrate of HRP, onto an immobilized protein. This system could be applied to analyses related to oxidase activity, as well as to the functional analysis of recombinant HRP.