International Journal of Molecular Sciences

1040 KiB

Open AccessArticle

Antiproliferative and Proapoptotic Activities of Methanolic Extracts from Ligustrum vulgare L. as an Individual Treatment and in Combination with Palladium Complex

by Milena G. Ćurčić, Milan S. Stanković, Emina M. Mrkalić, Zoran D. Matović, Dragić D. Banković, Danijela M. Cvetković, Dragana S. Đačić and Snežana D. Marković

Int. J. Mol. Sci. 2012, 13(2), 2521-2534; https://doi.org/10.3390/ijms13022521 - 22 Feb 2012

Cited by 54 | Viewed by 7798

Abstract

The aim of this study is to examine the growth inhibitory effects of methanolic leaf and fruit extracts of L. vulgare on HCT-116 cells over different time periods and their synergistic effect with a Pd(apox) complex. The antiproliferative activity of plant extracts alone [...] Read more.

The aim of this study is to examine the growth inhibitory effects of methanolic leaf and fruit extracts of L. vulgare on HCT-116 cells over different time periods and their synergistic effect with a Pd(apox) complex. The antiproliferative activity of plant extracts alone or in combination with the Pd(apox) complex was determined using MTT cell viability assay, where the IC₅₀ value was used as a parameter of cytotoxicity. Results show that antiproliferative effects of L. vulgare extracts increase with extension of exposure time, with decreasing IC₅₀ values, except for 72 h where the IC₅₀ values for methanolic leaf extract were lower than for the fruit extract. The Pd(apox) complex alone had a weak antiproliferative effect, but combination with L. vulgare extracts caused stronger effects with lower IC₅₀ values than with L. vulgare extracts alone. The type of cell death was explored by fluorescence microscopy using the acridin orange/ethidium bromide method. Treatments with plant extracts caused typical apoptotic morphological changes in HCT-116 cells and co-treatments with Pd(apox) complex caused higher levels of apoptotic cells than treatment with plant extracts alone. The results indicate that L. vulgare is a considerable source of natural bioactive substances with antiproliferative activity on HCT-116 cells and which have a substantial synergistic effect with the Pd(apox) complex. Full article

(This article belongs to the Section Biochemistry)

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88 KiB

Open AccessCommunication

Isolation and Characterization of New Microsatellite Markers for the Invasive Softshell Clam, Mya arenaria (L.) (Bivalvia: Myidae)

by Ana-Maria Krapal, Oana Paula Popa, Elena Iulia Iorgu, Marieta Costache and Luis Ovidiu Popa

Int. J. Mol. Sci. 2012, 13(2), 2515-2520; https://doi.org/10.3390/ijms13022515 - 22 Feb 2012

Cited by 4 | Viewed by 6280

Abstract

The invasive softshell clam (Mya arenaria Linnaeus, 1758) is native to the northwestern region of the Atlantic Ocean. This species has been introduced in the northeast Pacific and along the European coasts, due to intense naval transports and aquaculture, and it is [...] Read more.

The invasive softshell clam (Mya arenaria Linnaeus, 1758) is native to the northwestern region of the Atlantic Ocean. This species has been introduced in the northeast Pacific and along the European coasts, due to intense naval transports and aquaculture, and it is now present in all the European seas. In this paper we describe seven new microsatellite loci for Mya arenaria. The isolated loci are polymorphic with a number of alleles per locus between 6 and 14. The observed and expected heterozygosities ranged from 0.417 to 0.951, and from 0.643 to 0.895, with an average of 0.716 and 0.775, respectively. These microsatellite markers should be useful in analyzing this species’ genetic diversity, which could explain various processes of its invasion history. Full article

(This article belongs to the Section Biochemistry)

276 KiB

Open AccessArticle

Spectroscopic Parameter and Molecular Constant Investigations on Low-Lying States of BeF Radical

by Zun Lue Zhu, Qing Peng Song, Su Hua Kou, Jian Hua Lang and Jin Feng Sun

Int. J. Mol. Sci. 2012, 13(2), 2501-2514; https://doi.org/10.3390/ijms13022501 - 22 Feb 2012

Cited by 5 | Viewed by 6196

Abstract

The potential energy curves (PECs) of X²Σ⁺, A²Π_r and B²Σ⁺ states of BeF radical have been investigated using the complete active space self-consistent-field (CASSCF) method, followed by the highly accurate valence internally contracted [...] Read more.

The potential energy curves (PECs) of X²Σ⁺, A²Π_r and B²Σ⁺ states of BeF radical have been investigated using the complete active space self-consistent-field (CASSCF) method, followed by the highly accurate valence internally contracted multireference configuration interaction (MRCI) approach at the correlation-consistent basis sets, cc-pV5Z for Be and aug-cc-pV6Z for F. Based on the PECs of X²Σ⁺, A²Π_r and B²Σ⁺ states, the spectroscopic parameters (D_e, R_e, ω_e, ω_eχ_e, α_e and B_e) have also been determined in the present work. With the PECs determined at the present level of theory, vibrational states have been predicted for each state when the rotational quantum number J equals zero (J = 0). The vibrational levels, inertial rotation and centrifugal distortion constants are determined for the three states, and the classical turning points are also calculated for the X²Σ⁺ state. Compared with the available experiments and other theories, it can be seen that the present spectroscopic parameter and molecular constant results are more fully in agreement with the experimental findings. Full article

(This article belongs to the Special Issue Advances in Molecular Electronic Structure Calculations)

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2256 KiB

Open AccessArticle

The Expression Profiling of the Lipoxygenase (LOX) Family Genes During Fruit Development, Abiotic Stress and Hormonal Treatments in Cucumber (Cucumis sativus L.)

by Xue-Yong Yang, Wei-Jie Jiang and Hong-Jun Yu

Int. J. Mol. Sci. 2012, 13(2), 2481-2500; https://doi.org/10.3390/ijms13022481 - 22 Feb 2012

Cited by 96 | Viewed by 11733

Abstract

Lipoxygenases (LOXs) are non-haem iron-containing dioxygenases that catalyse oxygenation of polyunsaturated fatty acids and lipids to initiate the formation of a group of biologically active compounds called oxylipins. Plant oxylipins play important and diverse functions in the cells. In the current study, expression [...] Read more.

Lipoxygenases (LOXs) are non-haem iron-containing dioxygenases that catalyse oxygenation of polyunsaturated fatty acids and lipids to initiate the formation of a group of biologically active compounds called oxylipins. Plant oxylipins play important and diverse functions in the cells. In the current study, expression analysis during cucumber development using semi-quantitative RT-PCR revealed that 13 of 23 CsLOX genes were detectable, and were tissue specific or preferential accumulation. In total, 12 genes were found to be differentially expressed during fruit development and have different patterns of expression in exocarp, endocarp and pulp at day 5 after anthesis. The expression analysis of these 12 cucumber LOX genes in response to abiotic stresses and plant growth regulator treatments revealed their differential transcript in response to more than one treatment, indicating their diverse functions in abiotic stress and hormone responses. Results suggest that in cucumber the expanded LOX genes may play more diverse roles in life cycle and comprehensive data generated will be helpful in conducting functional genomic studies to understand their precise roles in cucumber fruit development and stress responses. Full article

(This article belongs to the Section Biochemistry)

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150 KiB

Open AccessArticle

Antioxidant Activity of Mulberry Fruit Extracts

by Muhammad Arfan, Rasool Khan, Anna Rybarczyk and Ryszard Amarowicz

Int. J. Mol. Sci. 2012, 13(2), 2472-2480; https://doi.org/10.3390/ijms13022472 - 22 Feb 2012

Cited by 103 | Viewed by 13334

Abstract

Phenolic compounds were extracted from the fruits of Morus nigra and Morus alba using methanol and acetone. The sugar-free extracts (SFEs) were prepared using Amberlite XAD-16 column chromatography. All of the SFEs exhibited antioxidant potential as determined by ABTS (0.75–1.25 mmol Trolox/g), DPPH [...] Read more.

Phenolic compounds were extracted from the fruits of Morus nigra and Morus alba using methanol and acetone. The sugar-free extracts (SFEs) were prepared using Amberlite XAD-16 column chromatography. All of the SFEs exhibited antioxidant potential as determined by ABTS (0.75–1.25 mmol Trolox/g), DPPH (2,2-diphenyl-1-picrylhydrazyl) (EC₅₀ from 48 μg/mL to 79 μg/mL), and reducing power assays. However, a stronger activity was noted for the SFEs obtained from Morus nigra fruits. These extracts also possessed the highest contents of total phenolics: 164 mg/g (methanolic SFE) and 173 mg/g (acetonic SFE). The presence of phenolic acids and flavonoids in the extracts was confirmed using HPLC method and chlorogenic acid and rutin were found as the dominant phenolic constituents in the SFEs. Full article

(This article belongs to the Special Issue Antioxidants)

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Graphical abstract

376 KiB

Open AccessArticle

First Insights on Organic Cosolvent Effects on FhuA Wildtype and FhuA Δ1-159

by Stefanie-Joana Tenne and Ulrich Schwaneberg

Int. J. Mol. Sci. 2012, 13(2), 2459-2471; https://doi.org/10.3390/ijms13022459 - 22 Feb 2012

Cited by 19 | Viewed by 6857

Abstract

Circular dichroism (CD) and deconvolution were used to study the structural integrity of a “plugged” and an “open” FhuA transmembrane channel protein in the presence of varied concentrations of tetrahydrofuran (THF), ethanol (EtOH) and chloroform/methanol (C/M). FhuA is an Escherichia coli outer membrane [...] Read more.

Circular dichroism (CD) and deconvolution were used to study the structural integrity of a “plugged” and an “open” FhuA transmembrane channel protein in the presence of varied concentrations of tetrahydrofuran (THF), ethanol (EtOH) and chloroform/methanol (C/M). FhuA is an Escherichia coli outer membrane protein (78.9 kDa) consisting of 22 β-sheets and an internal globular cork domain which acts as an iron transporter. FhuA and the deletion variant FhuA Δ1-159 showed comparable and remarkable resistance in the presence of THF (≤40 vol%) and EtOH (≤10 vol%). In C/M, significant differences in structural resistance were observed (FhuA stable ≤10 vol%; FhuA Δ1-159 ≤1 vol%). Deconvolution of CD-spectra for FhuA and FhuA Δ1-159 yielded β-sheet contents of 61 % (FhuA) and 58% (FhuA Δ1-159). Interestingly, FhuA and FhuA Δ1-159 had comparable β-sheet contents in the presence and absence of all three organic cosolvents. Additionally, precipitated FhuA and FhuA Δ1-159 (in 40 vol% C/M or 65 vol% THF) redissolved by supplementing the detergent n-octyl-oligo-oxyethylene (oPOE). Full article

(This article belongs to the Special Issue Applications of Circular Dichroism)

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Open AccessArticle

Enhancing Osteoconduction of PLLA-Based Nanocomposite Scaffolds for Bone Regeneration Using Different Biomimetic Signals to MSCs

by Gabriela Ciapetti, Donatella Granchi, Valentina Devescovi, Serena R. Baglio, Elisa Leonardi, Desirèe Martini, Maria Jesus Jurado, Beatriz Olalde, Ilaria Armentano, Josè M. Kenny, Frank X. Walboomers, Josè Inaki Alava and Nicola Baldini

Int. J. Mol. Sci. 2012, 13(2), 2439-2458; https://doi.org/10.3390/ijms13022439 - 22 Feb 2012

Cited by 36 | Viewed by 9936

Abstract

In bone engineering, the adhesion, proliferation and differentiation of mesenchymal stromal cells rely on signaling from chemico-physical structure of the substrate, therefore prompting the design of mimetic “extracellular matrix”-like scaffolds. In this study, three-dimensional porous poly-L-lactic acid (PLLA)-based scaffolds have been mixed with [...] Read more.

In bone engineering, the adhesion, proliferation and differentiation of mesenchymal stromal cells rely on signaling from chemico-physical structure of the substrate, therefore prompting the design of mimetic “extracellular matrix”-like scaffolds. In this study, three-dimensional porous poly-L-lactic acid (PLLA)-based scaffolds have been mixed with different components, including single walled carbon nanotubes (CNT), micro-hydroxyapatite particles (HA), and BMP2, and treated with plasma (PT), to obtain four different nanocomposites: PLLA + CNT, PLLA + CNTHA, PLLA + CNT + HA + BMP2 and PLLA + CNT + HA + PT. Adult bone marrow mesenchymal stromal cells (MSCs) were derived from the femur of orthopaedic patients, seeded on the scaffolds and cultured under osteogenic induction up to differentiation and mineralization. The release of specific metabolites and temporal gene expression profiles of marrow-derived osteoprogenitors were analyzed at definite time points, relevant to in vitro culture as well as in vivo differentiation. As a result, the role of the different biomimetic components added to the PLLA matrix was deciphered, with BMP2-added scaffolds showing the highest biomimetic activity on cells differentiating to mature osteoblasts. The modification of a polymeric scaffold with reinforcing components which also work as biomimetic cues for cells can effectively direct osteoprogenitor cells differentiation, so as to shorten the time required for mineralization. Full article

(This article belongs to the Special Issue Composite Materials in Skeletal Engineering)

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364 KiB

Open AccessArticle

DNA Barcodes of Asian Houbara Bustard (Chlamydotis undulata macqueenii)

by Ibrahim A. Arif, Haseeb A. Khan, Joseph B. Williams, Mohammad Shobrak and Waad I. Arif

Int. J. Mol. Sci. 2012, 13(2), 2425-2438; https://doi.org/10.3390/ijms13022425 - 22 Feb 2012

Cited by 5 | Viewed by 7949

Abstract

Populations of Houbara Bustards have dramatically declined in recent years. Captive breeding and reintroduction programs have had limited success in reviving population numbers and thus new technological solutions involving molecular methods are essential for the long term survival of this species. In this [...] Read more.

Populations of Houbara Bustards have dramatically declined in recent years. Captive breeding and reintroduction programs have had limited success in reviving population numbers and thus new technological solutions involving molecular methods are essential for the long term survival of this species. In this study, we sequenced the 694 bp segment of COI gene of the four specimens of Asian Houbara Bustard (Chlamydotis undulata macqueenii). We also compared these sequences with earlier published barcodes of 11 individuals comprising different families of the orders Gruiformes, Ciconiiformes, Podicipediformes and Crocodylia (out group). The pair-wise sequence comparison showed a total of 254 variable sites across all the 15 sequences from different taxa. Three of the four specimens of Houbara Bustard had an identical sequence of COI gene and one individual showed a single nucleotide difference (G > A transition at position 83). Within the bustard family (Otididae), comparison among the three species (Asian Houbara Bustard, Great Bustard (Otis tarda) and the Little Bustard (Tetrax tetrax)), representing three different genera, showed 116 variable sites. For another family (Rallidae), the intra-family variable sites among the individuals of four different genera were found to be 146. The COI genetic distances among the 15 individuals varied from 0.000 to 0.431. Phylogenetic analysis using 619 bp nucleotide segment of COI clearly discriminated all the species representing different genera, families and orders. All the four specimens of Houbara Bustard formed a single clade and are clearly separated from other two individuals of the same family (Otis tarda and Tetrax tetrax). The nucleotide sequence of partial segment of COI gene effectively discriminated the closely related species. This is the first study reporting the barcodes of Houbara Bustard and would be helpful in future molecular studies, particularly for the conservation of this threatened bird in Saudi Arabia. Full article

(This article belongs to the Section Biochemistry)

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318 KiB

Open AccessArticle

Characterization of Different Functionalized Lipidic Nanocapsules as Potential Drug Carriers

by Paola Sánchez-Moreno, Juan Luis Ortega-Vinuesa, Antonio Martín-Rodríguez, Houría Boulaiz, Juan Antonio Marchal-Corrales and José Manuel Peula-García

Int. J. Mol. Sci. 2012, 13(2), 2405-2424; https://doi.org/10.3390/ijms13022405 - 22 Feb 2012

Cited by 39 | Viewed by 9488

Abstract

Lipid nanocapsules (LNC) based on a core-shell structure consisting of an oil-filled core with a surrounding polymer layer are known to be promising vehicles for the delivery of hydrophobic drugs in the new therapeutic strategies in anti-cancer treatments. The present work has been [...] Read more.

Lipid nanocapsules (LNC) based on a core-shell structure consisting of an oil-filled core with a surrounding polymer layer are known to be promising vehicles for the delivery of hydrophobic drugs in the new therapeutic strategies in anti-cancer treatments. The present work has been designed as basic research about different LNC systems. We have synthesized—and physico-chemically characterized—three different LNC systems in which the core was constituted by olive oil and the shell by different phospholipids (phosphatidyl-serine or lecithin) and other biocompatible molecules such as Pluronic^® F68 or chitosan. It is notable that the olive-oil-phosphatidyl-serine LCN is a novel formulation presented in this work and was designed to generate an enriched carboxylic surface. This carboxylic layer is meant to link specific antibodies, which could facilitate the specific nanocapsule uptake by cancer cells. This is why nanoparticles with phosphatidyl-serine in their shell have also been used in this work to form immuno-nanocapsules containing a polyclonal IgG against a model antigen (C-reactive protein) covalently bounded by means of a simple and reproducible carbodiimide method. An immunological study was made to verify that these IgG-LNC complexes showed the expected specific immune response. Finally, a preliminary in vitro study was performed by culturing a breast-carcinoma cell line (MCF-7) with Nile-Red-loaded LNC. We found that these cancer cells take up the fluorescent Nile-Red molecule in a process dependent on the surface properties of the nanocarriers. Full article

(This article belongs to the Special Issue From Molecules to Nanomaterials)

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Graphical abstract

644 KiB

Open AccessArticle

Pyrrolo[3,2-d]pyrimidine Derivatives as Type II Kinase Insert Domain Receptor (KDR) Inhibitors: CoMFA and CoMSIA Studies

by Xiao-Yun Wu, Wen-Hua Chen, Shu-Guang Wu, Yuan-Xin Tian and Jia-Jie Zhang

Int. J. Mol. Sci. 2012, 13(2), 2387-2404; https://doi.org/10.3390/ijms13022387 - 22 Feb 2012

Cited by 11 | Viewed by 6833

Abstract

Kinase insert domain receptor (KDR) inhibitors have been proved to be very effective anticancer agents. Molecular docking, 3D-QSAR methods, CoMFA and CoMSIA were performed on pyrrolo[3,2-d]pyrimidine derivatives as non-ATP competitive KDR inhibitors (type II). The bioactive conformation was explored by docking [...] Read more.

Kinase insert domain receptor (KDR) inhibitors have been proved to be very effective anticancer agents. Molecular docking, 3D-QSAR methods, CoMFA and CoMSIA were performed on pyrrolo[3,2-d]pyrimidine derivatives as non-ATP competitive KDR inhibitors (type II). The bioactive conformation was explored by docking one potent compound 20 into the active site of KDR in its DFG-out inactive conformation. The constructed CoMFA and CoMSIA models produced statistically significant results with the cross-validated correlation coefficients q² of 0.542 and 0.552, non-cross-validated correlation coefficients r² of 0.912 and 0.955, and predicted correction coefficients r²_pred of 0.913 and 0.897, respectively. These results ensure the CoMFA and CoMSIA models as a tool to guide the design of a series of new potent KDR inhibitors. Full article

(This article belongs to the Section Physical Chemistry, Theoretical and Computational Chemistry)

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320 KiB

Open AccessReview

Impaired Iron Status in Aging Research

by Jinze Xu, Zhenhua Jia, Mitchell D. Knutson and Christiaan Leeuwenburgh

Int. J. Mol. Sci. 2012, 13(2), 2368-2386; https://doi.org/10.3390/ijms13022368 - 22 Feb 2012

Cited by 73 | Viewed by 10825

Abstract

Aging is associated with disturbances in iron metabolism and storage. During the last decade, remarkable progress has been made toward understanding their cellular and molecular mechanisms in aging and age-associated diseases using both cultured cells and animal models. The field has moved beyond [...] Read more.

Aging is associated with disturbances in iron metabolism and storage. During the last decade, remarkable progress has been made toward understanding their cellular and molecular mechanisms in aging and age-associated diseases using both cultured cells and animal models. The field has moved beyond descriptive studies to potential intervention studies focusing on iron chelation and removal. However, some findings remain controversial and inconsistent. This review summarizes important features of iron dyshomeostasis in aging research with a particular emphasis on current knowledge of the mechanisms underlying age-associated disorders in rodent models. Full article

(This article belongs to the Special Issue Oxidative Stress and Mitochondria)

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Graphical abstract

601 KiB

Open AccessArticle

Supercritical Carbon Dioxide Extraction of the Oak Silkworm (Antheraea pernyi) Pupal Oil: Process Optimization and Composition Determination

by Wen-Juan Pan, Ai-Mei Liao, Jian-Guo Zhang, Zeng Dong and Zhao-Jun Wei

Int. J. Mol. Sci. 2012, 13(2), 2354-2367; https://doi.org/10.3390/ijms13022354 - 21 Feb 2012

Cited by 38 | Viewed by 8489

Abstract

Supercritical carbon dioxide (SC-CO₂) extraction of oil from oak silkworm pupae was performed in the present research. Response surface methodology (RSM) was applied to optimize the parameters of SC-CO₂ extraction, including extraction pressure, temperature, time and CO₂ flow rate [...] Read more.

Supercritical carbon dioxide (SC-CO₂) extraction of oil from oak silkworm pupae was performed in the present research. Response surface methodology (RSM) was applied to optimize the parameters of SC-CO₂ extraction, including extraction pressure, temperature, time and CO₂ flow rate on the yield of oak silkworm pupal oil (OSPO). The optimal extraction condition for oil yield within the experimental range of the variables researched was at 28.03 MPa, 1.83 h, 35.31 °C and 20.26 L/h as flow rate of CO₂. Under this condition, the oil yield was predicted to be 26.18%. The oak silkworm pupal oil contains eight fatty acids, and is rich in unsaturated fatty acids and α-linolenic acid (ALA), accounting for 77.29% and 34.27% in the total oil respectively. Full article

(This article belongs to the Special Issue Supercritical Fluid Extraction)

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368 KiB

Open AccessReview

Epigenetic Disregulation in Oral Cancer

by Massimo Mascolo, Maria Siano, Gennaro Ilardi, Daniela Russo, Francesco Merolla, Gaetano De Rosa and Stefania Staibano

Int. J. Mol. Sci. 2012, 13(2), 2331-2353; https://doi.org/10.3390/ijms13022331 - 21 Feb 2012

Cited by 102 | Viewed by 11512

Abstract

Squamous cell carcinoma of the oral region (OSCC) is one of the most common and highly aggressive malignancies worldwide, despite the fact that significant results have been achieved during the last decades in its detection, prevention and treatment. Although many efforts have been [...] Read more.

Squamous cell carcinoma of the oral region (OSCC) is one of the most common and highly aggressive malignancies worldwide, despite the fact that significant results have been achieved during the last decades in its detection, prevention and treatment. Although many efforts have been made to define the molecular signatures that identify the clinical outcome of oral cancers, OSCC still lacks reliable prognostic molecular markers. Scientific evidence indicates that transition from normal epithelium to pre-malignancy, and finally to oral carcinoma, depends on the accumulation of genetic and epigenetic alterations in a multistep process. Unlike genetic alterations, epigenetic changes are heritable and potentially reversible. The most common examples of such changes are DNA methylation, histone modification, and small non-coding RNAs. Although several epigenetic changes have been currently linked to OSCC initiation and progression, they have been only partially characterized. Over the last decade, it has been demonstrated that especially aberrant DNA methylation plays a critical role in oral cancer. The major goal of the present paper is to review the recent literature about the epigenetic modifications contribution in early and later phases of OSCC malignant transformation; in particular we point out the current evidence of epigenetic marks as novel markers for early diagnosis and prognosis as well as potential therapeutic targets in oral cancer. Full article

(This article belongs to the Special Issue Advances in Molecular Pathology)

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Open AccessArticle

Antioxidant and Hepatoprotective Effects of the Red Ginseng Essential Oil in H₂O₂-Treated HepG2 Cells and CCl₄-Treated Mice

by Min-Ji Bak, Mira Jun and Woo-Sik Jeong

Int. J. Mol. Sci. 2012, 13(2), 2314-2330; https://doi.org/10.3390/ijms13022314 - 21 Feb 2012

Cited by 102 | Viewed by 12590

Abstract

The aim of this study was to evaluate the antioxidant mechanisms of red ginseng essential oil (REO) in cells as well as in an animal model. REO was prepared by a supercritical CO₂ extraction of waste-products generated after hot water extraction of [...] Read more.

The aim of this study was to evaluate the antioxidant mechanisms of red ginseng essential oil (REO) in cells as well as in an animal model. REO was prepared by a supercritical CO₂ extraction of waste-products generated after hot water extraction of red ginseng. In HepG2 cells, REO diminished the H₂O₂-mediated oxidative stress and also restored both the activity and expression of antioxidant enzymes such as superoxide dismutase, catalase and glutathione peroxidase. Administration of REO inhibited the phosphorylation of upstream mitogen-activated protein kinases (MAPKs) such as c-Jun N-terminal kinase, extracellular signal-regulated kinase, and p38. In mice, the CCl₄-mediated elevation of serum aspartate transaminase and alanine transaminase as well as the induction of hepatic lipid peroxidation were decreased by REO administration. REO treatments also resulted in up-regulation of the antioxidant enzyme expression in the liver. Moreover, increased phosphorylations of MAPKs were inhibited after REO administration. Overall, REO seems to protect the liver from oxidative stress through the activation and induction of antioxidant enzymes via inhibition of MAPKs pathways. Full article

(This article belongs to the Special Issue Advances in Nutraceutical Research)

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Open AccessArticle

Expression Patterns of Genes Involved in the Defense and Stress Response of Spiroplasma citri Infected Madagascar Periwinkle Catharanthus roseus

by Naghmeh Nejat, Ganesan Vadamalai and Matthew Dickinson

Int. J. Mol. Sci. 2012, 13(2), 2301-2313; https://doi.org/10.3390/ijms13022301 - 21 Feb 2012

Cited by 15 | Viewed by 6774

Abstract

Madagascar periwinkle is an ornamental and a medicinal plant, and is also an indicator plant that is highly susceptible to phytoplasma and spiroplasma infections from different crops. Periwinkle lethal yellows, caused by Spiroplasma citri, is one of the most devastating diseases of [...] Read more.

Madagascar periwinkle is an ornamental and a medicinal plant, and is also an indicator plant that is highly susceptible to phytoplasma and spiroplasma infections from different crops. Periwinkle lethal yellows, caused by Spiroplasma citri, is one of the most devastating diseases of periwinkle. The response of plants to S. citri infection is very little known at the transcriptome level. In this study, quantitative real-time PCR (RT-qPCR) was used to investigate the expression levels of four selected genes involved in defense and stress responses in naturally and experimentally Spiroplasma citri infected periwinkles. Strictosidine β-glucosidase involved in terpenoid indole alkaloids (TIAs) biosynthesis pathway showed significant upregulation in experimentally and naturally infected periwinkles. The transcript level of extensin increased in leaves of periwinkles experimentally infected by S. citri in comparison to healthy ones. A similar level of heat shock protein 90 and metallothionein expression was observed in healthy, naturally and experimentally spiroplasma-diseased periwinkles. Overexpression of Strictosidine β-glucosidase demonstrates the potential utility of this gene as a host biomarker to increase the fidelity of S. citri detection and can also be used in breeding programs to develop stable disease-resistance varieties. Full article

(This article belongs to the Section Molecular Pathology, Diagnostics, and Therapeutics)

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Open AccessArticle

In Vitro Control of Post-Harvest Fruit Rot Fungi by Some Plant Essential Oil Components

by Ippolito Camele, Luciana Altieri, Laura De Martino, Vincenzo De Feo, Emilia Mancini and Gian Luigi Rana

Int. J. Mol. Sci. 2012, 13(2), 2290-2300; https://doi.org/10.3390/ijms13022290 - 21 Feb 2012

Cited by 80 | Viewed by 10553

Abstract

Eight substances that are main components of the essential oils from three Mediterranean aromatic plants (Verbena officinalis, Thymus vulgaris and Origanum vulgare), previously found active against some phytopathogenic Fungi and Stramenopila, have been tested in vitro against five etiological agents [...] Read more.

Eight substances that are main components of the essential oils from three Mediterranean aromatic plants (Verbena officinalis, Thymus vulgaris and Origanum vulgare), previously found active against some phytopathogenic Fungi and Stramenopila, have been tested in vitro against five etiological agents of post-harvest fruit decay, Botrytis cinerea, Penicillium italicum, P. expansum, Phytophthora citrophthora and Rhizopus stolonifer. The tested compounds were β-fellandrene, β-pinene, camphene, carvacrol, citral, o-cymene, γ-terpinene and thymol. Citral exhibited a fungicidal action against P. citrophthora; carvacrol and thymol showed a fungistatic activity against P. citrophthora and R. stolonifer. Citral and carvacrol at 250 ppm, and thymol at 150 and 250 ppm stopped the growth of B. cinerea. Moreover, thymol showed fungistatic and fungicidal action against P. italicum. Finally, the mycelium growth of P. expansum was inhibited in the presence of 250 ppm of thymol and carvacrol. These results represent an important step toward the goal to use some essential oils or their components as natural preservatives for fruits and foodstuffs, due to their safety for consumer healthy and positive effect on shelf life extension of agricultural fresh products. Full article

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Open AccessArticle

Emodin Prevents Intrahepatic Fat Accumulation, Inflammation and Redox Status Imbalance During Diet-Induced Hepatosteatosis in Rats

by Anna Alisi, Anna Pastore, Sara Ceccarelli, Nadia Panera, Daniela Gnani, Giovannella Bruscalupi, Mara Massimi, Giulia Tozzi, Fiorella Piemonte and Valerio Nobili

Int. J. Mol. Sci. 2012, 13(2), 2276-2289; https://doi.org/10.3390/ijms13022276 - 20 Feb 2012

Cited by 54 | Viewed by 8584

Abstract

High-fat and/or high-carbohydrate diets may predispose to several metabolic disturbances including liver fatty infiltration (hepatosteatosis) or be associated with necro-inflammation and fibrosis (steatohepatitis). Several studies have emphasized the hepatoprotective effect of some natural agents. In this study, we investigated the potential therapeutic effects [...] Read more.

High-fat and/or high-carbohydrate diets may predispose to several metabolic disturbances including liver fatty infiltration (hepatosteatosis) or be associated with necro-inflammation and fibrosis (steatohepatitis). Several studies have emphasized the hepatoprotective effect of some natural agents. In this study, we investigated the potential therapeutic effects of the treatment with emodin, an anthraquinone derivative with anti-oxidant and anti-cancer abilities, in rats developing diet-induced hepatosteatosis and steatohepatitis. Sprague-Dawley rats were fed a standard diet (SD) for 15 weeks, or a high-fat/high-fructose diet (HFD/HF). After 5 weeks, emodin was added to the drinking water of some of the SD and HFD/HF rats. The experiment ended after an additional 10 weeks. Emodin-treated HFD/HF rats were protected from hepatosteatosis and metabolic derangements usually observed in HFD/HF animals. Furthermore, emodin exerted anti-inflammatory activity by inhibiting the HFD/HF-induced increase of tumor necrosis factor (TNF)-α. Emodin also affected the hepatocytes glutathione homeostasis and levels of the HFD/HF-induced increase of glutathionylated/phosphorylated phosphatase and tensin homolog (PTEN). In conclusion, we demonstrated that a natural agent such as emodin can prevent hepatosteatosis, preserving liver from pro-inflammatory and pro-oxidant damage caused by HFD/HF diet. These findings are promising, proposing emodin as a possible hindrance to progression of hepatosteatosis into steatohepatitis. Full article

(This article belongs to the Special Issue Antioxidants)

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Open AccessArticle

Comprehensive and Facile Synthesis of Some Functionalized Bis-Heterocyclic Compounds Containing a Thieno[2,3-b]thiophene Motif

by Yahia N. Mabkhot, Assem Barakat, Abdullah M. Al-Majid and Saeed A. Alshahrani

Int. J. Mol. Sci. 2012, 13(2), 2263-2275; https://doi.org/10.3390/ijms13022263 - 20 Feb 2012

Cited by 21 | Viewed by 6839

Abstract

A comprehensive and facile method for the synthesis of new functionalized bis-heterocyclic compounds containing a thieno[2,3-b]thiophene motif is described. The hitherto unknown bis-pyrazolothieno[2,3-b]thiophene derivatives 2a–c, bis-pyridazin othieno[2,3-b]thiophene derivatives 4, bis-pyridinothieno[2,3-b]thiophene derivatives [...] Read more.

A comprehensive and facile method for the synthesis of new functionalized bis-heterocyclic compounds containing a thieno[2,3-b]thiophene motif is described. The hitherto unknown bis-pyrazolothieno[2,3-b]thiophene derivatives 2a–c, bis-pyridazin othieno[2,3-b]thiophene derivatives 4, bis-pyridinothieno[2,3-b]thiophene derivatives 6a,b, and to an analogous bis-pyridinothieno[2,3-b]thiophene nitrile derivatives 7 are obtained. Additionally, the novel bis-pyradazinonothieno[2,3-b]thiophene derivatives 9, and nicotinic acid derivatives 10, 11 are obtained via bis-dienamide 8. The structures of all newly synthesized compounds have been elucidated by ¹H, ¹³C NMR, GCMS, and IR spectrometry. These compounds represent a new class of sulfur and Nitrogen containing heterocycles that should also be of interest as new materials. Full article

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Open AccessArticle

Seasonal Variations of the Antioxidant Composition in Ground Bamboo Sasa argenteastriatus Leaves

by Qinxue Ni, Guangzhi Xu, Zhiqiang Wang, Qianxin Gao, Shu Wang and Youzuo Zhang

Int. J. Mol. Sci. 2012, 13(2), 2249-2262; https://doi.org/10.3390/ijms13022249 - 20 Feb 2012

Cited by 45 | Viewed by 8285

Abstract

Sasa argenteastriatus, with abundant active compounds and high antioxidant activity in leaves, is a new leafy bamboo grove suitable for exploitation. To utilize it more effectively and scientifically, we investigate the seasonal variations of antioxidant composition in its leaves and antioxidant activity. [...] Read more.

Sasa argenteastriatus, with abundant active compounds and high antioxidant activity in leaves, is a new leafy bamboo grove suitable for exploitation. To utilize it more effectively and scientifically, we investigate the seasonal variations of antioxidant composition in its leaves and antioxidant activity. The leaves of Sasa argenteastriatus were collected on the 5th day of each month in three same-sized sample plots from May 2009 to May 2011. The total flavonoids (TF): phenolics (TP) and triterpenoid (TT) of bamboo leaves were extracted and the contents analyzed by UV-spectrophotometer. Our data showed that all exhibited variations with the changing seasons, with the highest levels appearing in November to March. Antioxidant activity was measured using DPPH and FRAP methods. The highest antioxidant activity appeared in December with the lowest in May. Correlation analyses demonstrated that TP and TF exhibited high correlation with bamboo antioxidant activity. Eight bamboo characteristic compounds (orientin, isoorientin, vitexin, homovitexin and p-coumaric acid, chlorogenic acid, caffeic acid, ferulic acid) were determined by RP-HPLC synchronously. We found that chlorogenic acid, isoorientin and vitexin are the main compounds in Sasa argenteastriatus leaves and the content of isovitexin and chlorogenic acid showed a similar seasonal variation with the TF, TP and TT. Our results suggested that the optimum season for harvesting Sasa argenteastriatus leaves is between autumn and winter. Full article

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Open AccessArticle

Nanosecond T-Jump Experiment in Poly(glutamic acid): A Circular Dichroism Study

by Lucille Mendonça and François Hache

Int. J. Mol. Sci. 2012, 13(2), 2239-2248; https://doi.org/10.3390/ijms13022239 - 17 Feb 2012

Cited by 15 | Viewed by 7962

Abstract

Poly(glutamic acid) has been studied with a nanosecond T-jump experiment. A new experimental set-up based on the frequency-quadrupling of an 82 MHz Titanium-Sapphire laser allows rapid CD measurements to be performed. Combining time-resolved absorption and circular dichroism at 204 and 220 nm, we [...] Read more.

Poly(glutamic acid) has been studied with a nanosecond T-jump experiment. A new experimental set-up based on the frequency-quadrupling of an 82 MHz Titanium-Sapphire laser allows rapid CD measurements to be performed. Combining time-resolved absorption and circular dichroism at 204 and 220 nm, we are able to measure precisely the unfolding relaxation time as well as the helical fraction evolution. We show that only CD at 220 nm is relevant to observe the unfolding of an alpha helix whereas no change is observed for CD at 204 nm. Conversely, both absorptions yield information on the dynamics of the process. Full article

(This article belongs to the Special Issue Applications of Circular Dichroism)

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Open AccessReview

Alpha7 Nicotinic Acetylcholine Receptor Is a Target in Pharmacology and Toxicology

by Miroslav Pohanka

Int. J. Mol. Sci. 2012, 13(2), 2219-2238; https://doi.org/10.3390/ijms13022219 - 17 Feb 2012

Cited by 153 | Viewed by 17936

Abstract

Alpha7 nicotinic acetylcholine receptor (α7 nAChR) is an important part of the cholinergic nerve system in the brain. Moreover, it is associated with a cholinergic anti-inflammatory pathway in the termination of the parasympathetic nervous system. Antagonists of α7 nAChR are a wide group [...] Read more.

Alpha7 nicotinic acetylcholine receptor (α7 nAChR) is an important part of the cholinergic nerve system in the brain. Moreover, it is associated with a cholinergic anti-inflammatory pathway in the termination of the parasympathetic nervous system. Antagonists of α7 nAChR are a wide group represented by conotoxin and bungarotoxin. Even Alzheimer’s disease drug memantine acting as an antagonist in its side pathway belongs in this group. Agonists of α7 nAChR are suitable for treatment of multiple cognitive dysfunctions such as Alzheimer’s disease or schizophrenia. Inflammation or even sepsis can be ameliorated by the agonistic acting compounds. Preparations RG3487, SEN34625/WYE-103914, SEN12333, ABT-107, Clozapine, GTS-21, CNI-1493, and AR-R17779 are representative examples of the novel compounds with affinity toward the α7 nAChR. Pharmacological, toxicological, and medicinal significance of α7 nAChR are discussed throughout this paper. Full article

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A Novel LMP1 Antibody Synergizes with Mitomycin C to Inhibit Nasopharyngeal Carcinoma Growth in Vivo Through Inducing Apoptosis and Downregulating Vascular Endothelial Growth Factor

by Yuan Mao, Da-Wei Zhang, Juan Wen, Qing Cao, Ren-Jie Chen, Jin Zhu and Zhen-Qing Feng

Int. J. Mol. Sci. 2012, 13(2), 2208-2218; https://doi.org/10.3390/ijms13022208 - 17 Feb 2012

Cited by 22 | Viewed by 8121

Abstract

Combined therapy emerges as an attractive strategy for cancer treatment. The aim of this study was to investigate the inhibitory effects of mitomycin C (MMC) combined with a novel antibody fragment (Fab) targeting latent membrane protein 1 (LMP1) on nasopharyngeal carcinoma (NPC) xenograft [...] Read more.

Combined therapy emerges as an attractive strategy for cancer treatment. The aim of this study was to investigate the inhibitory effects of mitomycin C (MMC) combined with a novel antibody fragment (Fab) targeting latent membrane protein 1 (LMP1) on nasopharyngeal carcinoma (NPC) xenograft nude mice. The inhibitory rates of MMC (2 mg/kg), Fab (4 mg/kg), MMC (2 mg/kg) + Fab (4 mg/kg), and MMC (1 mg/kg) + Fab (4 mg/kg) were 20.1%, 7.3%, 42.5% and 40.5%, respectively. Flow cytometry analysis showed that the apoptotic rate of xenograft tumor cells in the MMC and Fab combination group was 28 ± 4.12%, significantly higher than the MMC (2 mg/kg) group (P < 0.01). Immunohistochemical staining showed that VEGF expression in NPC xenografts was significantly inhibited in the combination group compared to the Fab (4 mg/kg) group (P < 0.05). In conclusion, both MMC and Fab could inhibit NPC xenograft tumor growth in vivo and combination therapy showed apparent synergistic anti-tumor effects, which may be due to the induction of tumor cell apoptosis and the downregulation of VEGF expression. These results suggest that the novel combined therapy utilizing traditional chemotherapeutics and antibody-targeted therapy could be a promising strategy for the treatment of NPC. Full article

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Open AccessArticle

Using Support Vector Machine and Evolutionary Profiles to Predict Antifreeze Protein Sequences

by Xiaowei Zhao, Zhiqiang Ma and Minghao Yin

Int. J. Mol. Sci. 2012, 13(2), 2196-2207; https://doi.org/10.3390/ijms13022196 - 17 Feb 2012

Cited by 44 | Viewed by 7253

Abstract

Antifreeze proteins (AFPs) are ice-binding proteins. Accurate identification of new AFPs is important in understanding ice-protein interactions and creating novel ice-binding domains in other proteins. In this paper, an accurate method, called AFP_PSSM, has been developed for predicting antifreeze proteins using a support [...] Read more.

Antifreeze proteins (AFPs) are ice-binding proteins. Accurate identification of new AFPs is important in understanding ice-protein interactions and creating novel ice-binding domains in other proteins. In this paper, an accurate method, called AFP_PSSM, has been developed for predicting antifreeze proteins using a support vector machine (SVM) and position specific scoring matrix (PSSM) profiles. This is the first study in which evolutionary information in the form of PSSM profiles has been successfully used for predicting antifreeze proteins. Tested by 10-fold cross validation and independent test, the accuracy of the proposed method reaches 82.67% for the training dataset and 93.01% for the testing dataset, respectively. These results indicate that our predictor is a useful tool for predicting antifreeze proteins. A web server (AFP_PSSM) that implements the proposed predictor is freely available. Full article

(This article belongs to the Section Biochemistry)

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Computational Studies of Difference in Binding Modes of Peptide and Non-Peptide Inhibitors to MDM2/MDMX Based on Molecular Dynamics Simulations

by Jianzhong Chen, Dinglin Zhang, Yuxin Zhang and Guohui Li

Int. J. Mol. Sci. 2012, 13(2), 2176-2195; https://doi.org/10.3390/ijms13022176 - 17 Feb 2012

Cited by 24 | Viewed by 8558

Abstract

Inhibition of p53-MDM2/MDMX interaction is considered to be a promising strategy for anticancer drug design to activate wild-type p53 in tumors. We carry out molecular dynamics (MD) simulations to study the binding mechanisms of peptide and non-peptide inhibitors to MDM2/MDMX. The rank of [...] Read more.

Inhibition of p53-MDM2/MDMX interaction is considered to be a promising strategy for anticancer drug design to activate wild-type p53 in tumors. We carry out molecular dynamics (MD) simulations to study the binding mechanisms of peptide and non-peptide inhibitors to MDM2/MDMX. The rank of binding free energies calculated by molecular mechanics generalized Born surface area (MM-GBSA) method agrees with one of the experimental values. The results suggest that van der Waals energy drives two kinds of inhibitors to MDM2/MDMX. We also find that the peptide inhibitors can produce more interaction contacts with MDM2/MDMX than the non-peptide inhibitors. Binding mode predictions based on the inhibitor-residue interactions show that the π–π, CH–π and CH–CH interactions dominated by shape complimentarity, govern the binding of the inhibitors in the hydrophobic cleft of MDM2/MDMX. Our studies confirm the residue Tyr99 in MDMX can generate a steric clash with the inhibitors due to energy and structure. This finding may theoretically provide help to develop potent dual-specific or MDMX inhibitors. Full article

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Open AccessArticle

On the Electrophilic Character of Molecules Through Its Relation with Electronegativity and Chemical Hardness

by Nazmul Islam and Dulal C. Ghosh

Int. J. Mol. Sci. 2012, 13(2), 2160-2175; https://doi.org/10.3390/ijms13022160 - 17 Feb 2012

Cited by 42 | Viewed by 7638

Abstract

Electrophilicity is an intrinsic property of atoms and molecules. It probably originates logistically with the involvement in the physical process of electrostatics of soaked charge in electronic shells and the screened nuclear charge of atoms. Motivated by the existing view of conceptual density [...] Read more.

Electrophilicity is an intrinsic property of atoms and molecules. It probably originates logistically with the involvement in the physical process of electrostatics of soaked charge in electronic shells and the screened nuclear charge of atoms. Motivated by the existing view of conceptual density functional theory that similar to electronegativity and hardness equalization, there should be a physical process of equalization of electrophilicity during the chemical process of formation of hetero nuclear molecules, we have developed a new theoretical scheme and formula for evaluating the electrophilicity of hetero nuclear molecules. A comparative study with available bench marking reveals that the hypothesis of electrophilicity and equalization, and the present method of evaluating equalized electrophilicity, are scientifically promising. Full article

(This article belongs to the Special Issue Atoms in Molecules and in Nanostructures)

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Open AccessArticle

Benzyl and Methyl Fatty Hydroxamic Acids Based on Palm Kernel Oil as Chelating Agent for Liquid-Liquid Iron(III) Extraction

by Md Jelas Haron, Hossein Jahangirian, Sidik Silong, Nor Azah Yusof, Anuar Kassim, Roshanak Rafiee-Moghaddam, Behnam Mahdavi, Mazyar Peyda, Yadollah Abdollahi and Jamileh Amin

Int. J. Mol. Sci. 2012, 13(2), 2148-2159; https://doi.org/10.3390/ijms13022148 - 16 Feb 2012

Cited by 9 | Viewed by 7266

Abstract

Liquid-liquid iron(III) extraction was investigated using benzyl fatty hydroxamic acids (BFHAs) and methyl fatty hydroxamic acids (MFHAs) as chelating agents through the formation of iron(III) methyl fatty hydroxamate (Fe-MFHs) or iron(III) benzyl fatty hydroxamate (Fe-BFHs) in the organic phase. The results obtained under [...] Read more.

Liquid-liquid iron(III) extraction was investigated using benzyl fatty hydroxamic acids (BFHAs) and methyl fatty hydroxamic acids (MFHAs) as chelating agents through the formation of iron(III) methyl fatty hydroxamate (Fe-MFHs) or iron(III) benzyl fatty hydroxamate (Fe-BFHs) in the organic phase. The results obtained under optimized conditions, showed that the chelating agents in hexane extract iron(III) at pH 1.9 were realized effectively with a high percentage of extraction (97.2% and 98.1% for MFHAs and BFHAs, respectively). The presence of a large amount of Mg(II), Ni(II), Al(III), Mn(II) and Co(II) ions did affect the iron(III) extraction. Finally stripping studies for recovering iron(III) from organic phase (Fe-MFHs or Fe-BFHs dissolved in hexane) were carried out at various concentrations of HCl, HNO₃and H₂SO₄. The results showed that the desired acid for recovery of iron(III) was 5 M HCl and quantitative recovery of iron(III) was achieved from Fe(III)-MFHs and Fe(III)-BFHs solutions in hexane containing 5 mg/L of Fe(III). Full article

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cDNA Cloning, Overexpression, Purification and Pharmacologic Evaluation for Anticancer Activity of Ribosomal Protein L23A Gene (RPL23A) from the Giant Panda

by Bing Sun, Yi-Ling Hou, Wan-Ru Hou, Si-Nan Zhang, Xiang Ding and Xiu-Lan Su

Int. J. Mol. Sci. 2012, 13(2), 2133-2147; https://doi.org/10.3390/ijms13022133 - 16 Feb 2012

Cited by 13 | Viewed by 8587

Abstract

RPL23A gene encodes a ribosomal protein that is a component of the 60S subunit. The protein belongs to the L23P family of ribosomal proteins, which is located in the cytoplasm. The purpose of this paper was to explore the structure and anti-cancer function [...] Read more.

RPL23A gene encodes a ribosomal protein that is a component of the 60S subunit. The protein belongs to the L23P family of ribosomal proteins, which is located in the cytoplasm. The purpose of this paper was to explore the structure and anti-cancer function of ribosomal protein L23A (RPL23A) gene of the Giant Panda (Ailuropoda melanoleuca). The cDNA of RPL23A was cloned successfully from the Giant Panda using RT-PCR technology. We constructed a recombinant expression vector containing RPL23A cDNA and over-expressed it in Escherichia coli using pET28a plasmids. The expression product obtained was purified by using Ni chelating affinity chromatography. Recombinant protein of RPL23A obtained from the experiment acted on Hep-2 cells and human HepG-2 cells, then the growth inhibitory effect of these cells was observed by MTT (3-[4,5-dimethyl-2-thiazolyl]-2,5-diphenyl-2H-tetrazolium bromide) assay. The result indicated that the length of the fragment cloned is 506 bp, and it contains an open-reading frame (ORF) of 471 bp encoding 156 amino acids. Primary structure analysis revealed that the molecular weight of the putative RPL23A protein is 17.719 kDa with a theoretical pI 11.16. The molecular weight of the recombinant protein RPL23A is 21.265 kDa with a theoretical pI 10.57. The RPL23A gene can be really expressed in E. coli and the RPL23A protein, fusioned with the N-terminally His-tagged protein, gave rise to the accumulation of an expected 22 KDa polypeptide. The data showed that the recombinant protein RPL23A had a time- and dose-dependency on the cell growth inhibition rate. The data also indicated that the effect at low concentrations was better than at high concentrations on Hep-2 cells, and that the concentration of 0.185 μg/mL had the best rate of growth inhibition of 36.31%. All results of the experiment revealed that the recombinant protein RPL23A exhibited anti-cancer function on the Hep-2 cells. The study provides a scientific basis and aids orientation for the research and development of cancer protein drugs as well as possible anti-cancer mechanisms. Further research is on going to determine the bioactive principle(s) of recombinant protein RPL23A responsible for its anticancer activity. Full article

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Open AccessArticle

Correlation Coefficients Between Different Methods of Expressing Bacterial Quantification Using Real Time PCR

by Bahman Navidshad, Juan Boo Liang and Mohammad Faseleh Jahromi

Int. J. Mol. Sci. 2012, 13(2), 2119-2132; https://doi.org/10.3390/ijms13022119 - 16 Feb 2012

Cited by 42 | Viewed by 8435

Abstract

The applications of conventional culture-dependent assays to quantify bacteria populations are limited by their dependence on the inconsistent success of the different culture-steps involved. In addition, some bacteria can be pathogenic or a source of endotoxins and pose a health risk to the [...] Read more.

The applications of conventional culture-dependent assays to quantify bacteria populations are limited by their dependence on the inconsistent success of the different culture-steps involved. In addition, some bacteria can be pathogenic or a source of endotoxins and pose a health risk to the researchers. Bacterial quantification based on the real-time PCR method can overcome the above-mentioned problems. However, the quantification of bacteria using this approach is commonly expressed as absolute quantities even though the composition of samples (like those of digesta) can vary widely; thus, the final results may be affected if the samples are not properly homogenized, especially when multiple samples are to be pooled together before DNA extraction. The objective of this study was to determine the correlation coefficients between four different methods of expressing the output data of real-time PCR-based bacterial quantification. The four methods were: (i) the common absolute method expressed as the cell number of specific bacteria per gram of digesta; (ii) the Livak and Schmittgen, ΔΔCt method; (iii) the Pfaffl equation; and (iv) a simple relative method based on the ratio of cell number of specific bacteria to the total bacterial cells. Because of the effect on total bacteria population in the results obtained using ΔCt-based methods (ΔΔCt and Pfaffl), these methods lack the acceptable consistency to be used as valid and reliable methods in real-time PCR-based bacterial quantification studies. On the other hand, because of the variable compositions of digesta samples, a simple ratio of cell number of specific bacteria to the corresponding total bacterial cells of the same sample can be a more accurate method to quantify the population. Full article

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The Effect of Toll-Like Receptor 4 in the Aqueous Humor of Endotoxin-Induced Uveitis

by Jing Wang, Hong Lu, Yingzhi Xu, Xiaofeng Hu, Wei Chen and Shang Li

Int. J. Mol. Sci. 2012, 13(2), 2110-2118; https://doi.org/10.3390/ijms13022110 - 16 Feb 2012

Cited by 5 | Viewed by 6344

Abstract

In our previous study, we found that acute anterior uveitis (AAU) could be induced in wild-type mice (C3H/HeN), but it could not be induced in TLR4 gene-deficient mice (C3H/HeJ), we concluded that the translocation of transcription factor nuclear factor-κB (NF-κB) may play an [...] Read more.

In our previous study, we found that acute anterior uveitis (AAU) could be induced in wild-type mice (C3H/HeN), but it could not be induced in TLR4 gene-deficient mice (C3H/HeJ), we concluded that the translocation of transcription factor nuclear factor-κB (NF-κB) may play an important role. In this study, we examined the concentration of different cytokines in the aqueous humor of C3H/HeN mice and C3H/HeJ mice with the aim of exploring the role of different cytokines in the lipopolysaccharide (LPS) and TLR4-mediated signal transduction in the development of AAU. Full article

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Open AccessReview

Antioxidant-Induced Stress

by Cleva Villanueva and Robert D. Kross

Int. J. Mol. Sci. 2012, 13(2), 2091-2109; https://doi.org/10.3390/ijms13022091 - 16 Feb 2012

Cited by 115 | Viewed by 12822

Abstract

Antioxidants are among the most popular health-protecting products, sold worldwide without prescription. Indeed, there are many reports showing the benefits of antioxidants but only a few questioning the possible harmful effects of these “drugs”. The normal balance between antioxidants and free radicals in [...] Read more.

Antioxidants are among the most popular health-protecting products, sold worldwide without prescription. Indeed, there are many reports showing the benefits of antioxidants but only a few questioning the possible harmful effects of these “drugs”. The normal balance between antioxidants and free radicals in the body is offset when either of these forces prevails. The available evidence on the harmful effects of antioxidants is analyzed in this review. In summary, a hypothesis is presented that “antioxidant-induced stress” results when antioxidants overwhelm the body’s free radicals. Full article

(This article belongs to the Special Issue Bioactive Nanoparticles (special issue))

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Int. J. Mol. Sci., Volume 13, Issue 2 (February 2012) – 80 articles , Pages 1269-2534

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