IJMS: Molecular Pathology

IJMS, Vol. 11, Pages 544-561: Novel Neuroprotective Strategies in Ischemic Retinal Lesions

Krisztina Szabadfi — Wed, 03 Feb 2010 00:00:00 CET

Retinal ischemia can be effectively modeled by permanent bilateral common carotid artery occlusion, which leads to chronic hypoperfusion-induced degeneration in the entire rat retina. The complex pathways leading to retinal cell death offer a complex approach of neuroprotective strategies. In the present review we summarize recent findings with different neuroprotective candidate molecules. We describe the protective effects of intravitreal treatment with: (i) urocortin 2; (ii) a mitochondrial ATP-sensitive K+ channel opener, diazoxide; (iii) a neurotrophic factor, pituitary adenylate cyclase activating polypeptide; and (iv) a novel poly(ADP-ribose) polymerase inhibitor (HO3089). The retinoprotective effects are demonstrated with morphological description and effects on apoptotic pathways using molecular biological techniques.

IJMS, Vol. 11, Pages 312-328: Melatonin and Structurally-Related Compounds Protect Synaptosomal Membranes from Free Radical Damage

Sergio Millán-Plano — Thu, 21 Jan 2010 00:00:00 CET

Since biological membranes are composed of lipids and proteins we tested the in vitro antioxidant properties of several indoleamines from the tryptophan metabolic pathway in the pineal gland against oxidative damage to lipids and proteins of synaptosomes isolated from the rat brain. Free radicals were generated by incubation with 0.1 mM FeCl3, and 0.1 mM ascorbic acid. Levels of malondialdehyde (MDA) plus 4-hydroxyalkenal (4-HDA), and carbonyl content in the proteins were measured as indices of oxidative damage to lipids and proteins, respectively. Pinoline was the most powerful antioxidant evaluated, with melatonin, N-acetylserotonin, 5-hydroxytryptophan, 5-methoxytryptamine, 5-methoxytryptophol, and tryptoline also acting as antioxidants.

IJMS, Vol. 11, Pages 140-153: Neuroprotective Effects of Ischemic Preconditioning on Global Brain Ischemia through Up-Regulation of Acid-Sensing Ion Channel 2a

Yifeng Miao — Tue, 12 Jan 2010 00:00:00 CET

Transient forebrain or global ischemia induces cell death in vulnerable CA1 pyramidal neurons. A brief period of ischemia, i.e., ischemic preconditioning, affords CA1 neurons robust protection against a subsequent, more prolonged ischemic challenge. Using the four-vessel occlusion model, we established an ischemic preconditioning model in which rodents were subjected to 3 min of sublethal ischemia 48 h before a 15 min lethal ischemia. We showed that preconditioning attenuated the ischemia-induced neural cell death and DNA fragmentation in the hippocampal CA1 region. RT-PCR and western blot analysis showed that preconditioning prior to an ischemic insult significantly increased ASIC 2a mRNA and protein expression in comparison to the ischemic insult alone (p < 0.01). These findings implicate a new role of ASIC 2a on endogenous neuroprotection from ischemic insult.

IJMS, Vol. 11, Pages 107-123: Ginkgo biloba Extract in Alzheimer’s Disease: From Action Mechanisms to Medical Practice

Chun Shi — Fri, 08 Jan 2010 00:00:00 CET

Standardized extract from the leaves of the Ginkgo biloba tree, labeled EGb761, is one of the most popular herbal supplements. Numerous preclinical studies have shown the neuroprotective effects of EGb761 and support the notion that it may be effective in the treatment and prevention of neurodegenerative disorders such as Alzheimer’s disease (AD). Despite the preclinical promise, the clinical efficacy of this drug remains elusive. In this review, possible mechanisms underlying neuroprotective actions of EGb761 are described in detail, together with a brief discussion of the problem of studying this herb clinically to verify its efficacy in the treatment and prevention of AD. Moreover, various parameters e.g., the dosage and the permeability of the blood brain barrier (BBB), impacting the outcome of the clinical effectiveness of the extract are also discussed. Overall, the findings summarized in this review suggest that, a better understanding of the neuroprotective mechanisms of EGb761 may contribute to better understanding of the effectiveness and complexity of this herb and may also be helpful for design of therapeutic strategies in future clinical practice. Therefore, in future clinical studies, different factors that could interfere with the effect of EGb761 should be considered.

IJMS, Vol. 10, Pages 5471-5484: Immune Response among Patients Exposed to Molds

David A. Edmondson — Mon, 21 Dec 2009 00:00:00 CET

Macrocyclic trichothecenes, mycotoxins produced by Stachybotrys chartarum, have been implicated in adverse reactions in individuals exposed to mold-contaminated environments. Cellular and humoral immune responses and the presence of trichothecenes were evaluated in patients with mold-related health complaints. Patients underwent history, physical examination, skin prick/puncture tests with mold extracts, immunological evaluations and their sera were analyzed for trichothecenes. T-cell proliferation, macrocyclic trichothecenes, and mold specific IgG and IgA levels were not significantly different than controls; however 70% of the patients had positive skin tests to molds. Thus, IgE mediated or other non-immune mechanisms could be the cause of their symptoms.

IJMS, Vol. 10, Pages 5090-5103: Mycoflora and Co-Occurrence of Fumonisins and Aflatoxins in Freshly Harvested Corn in Different Regions of Brazil

Liliana O. Rocha — Tue, 24 Nov 2009 00:00:00 CET

Natural mycoflora and co-occurrence of fumonisins (FB1, FB2) and aflatoxins (AFB1, AFB2, AFG1 and AFG2) in freshly harvested corn grain samples from four regions of Brazil were investigated. Fusarium verticillioides was predominant in all samples. Analysis of fumonisins showed that 98% of the samples were contaminated with FB1 and 74.5% with FB1 + FB2, with toxin levels ranging from 0.015 to 9.67 μg/g for FB1 and from 0.015 to 3.16 μg/g for FB2. Twenty-one (10.5%) samples were contaminated with AFB1, seven (3.5%) with AFB2 and only one (0.5%) with AFG1 and AFG2. Co-contamination with aflatoxins and fumonisins was observed in 7% of the samples. The highest contamination of fumonisins and aflatoxins was observed in Nova Odessa (SP) and Várzea Grande (MT), respectively. The lowest contamination of these mycotoxins was found in Várzea Grande and Nova Odessa, respectively.

IJMS, Vol. 10, Pages 4789-4804: Oxidative Neurodegeneration Is Prevented by UCP0045037, an Allosteric Modulator for the Reduced Form of DJ-1, a Wild-Type of Familial Parkinson’s Disease-Linked PARK7

Koichiro Yamane — Thu, 05 Nov 2009 00:00:00 CET

Although a loss-of-function mutation has been identified in familial Parkinson’s disease PARK7, the wild-type of DJ-1 is known to act as an oxidative stress sensor in neuronal cells. Recently, we identified UCP0045037 as a compound that bound to the reduced form of DJ-1 by in silico virtual screening. In this study, we determined the neuroprotective effects of UCP0045037 against focal cerebral ischemia-induced neurodegeneration in rats. Hydrogen peroxide-induced cell death was significantly inhibited by UCP0045037 in both rat mesencephalic dopaminergic neurons and human normal SH-SY5Y cells. In contrast, DJ-1-knockdown SH-SY5Y cells lost the protective activity of UCP0045037. These results suggest that UCP0045037 interacts with endogenous DJ-1 and produces a neuroprotective response.

IJMS, Vol. 10, Pages 3900-3917: Expression of CD40 and CD40L in Gastric Cancer Tissue and Its Clinical Significance

Rui Li — Fri, 04 Sep 2009 00:00:00 CEST

To study expression of CD40 and CD40L in gastric cancer tissue we assessed gastric cancer patients admitted to the Department of Gastroenterology of The First Affiliated Hospital of Soochow University and control subjects. Gastric cancer and normal (from around tumours) tissue samples were obtained from patients. Venous blood samples (gastric cancer and ulcer groups) were drawn on the morning of the day before surgery for the measurement of peripheral sCD40L. The expression of CD40 in gastric carcinoma specimens was examined immuno-histochemically. The clinicopathological factors, including age, sex, tumor size, gross appearance, degree of cellular differentiation, histological classification, depth of tumor invasion, lymph node metastasis, peritoneal dissemination, and TNM stage were analyzed according to the different expression of CD40. The results indicated a high CD40 expression in gastric cancer tissues. This positive expression of CD40 revealed a significant (P < 0.05) correlation with lymphatic metastasis and tumor TNM stage in gastric cancer patients. It is concluded that higher CD40 expression existed in expanding type tumors and could play an important role in clinical diagnosis of gastric cancer patients.

IJMS, Vol. 10, Pages 2510-2557: Trends in the Molecular Pathogenesis and Clinical Therapeutics of Common Neurodegenerative Disorders

Yahya E. Choonara — Wed, 03 Jun 2009 00:00:00 CEST

The term neurodegenerative disorders, encompasses a variety of underlying conditions, sporadic and/or familial and are characterized by the persistent loss of neuronal subtypes. These disorders can disrupt molecular pathways, synapses, neuronal subpopulations and local circuits in specific brain regions, as well as higher-order neural networks. Abnormal network activities may result in a vicious cycle, further impairing the integrity and functions of neurons and synapses, for example, through aberrant excitation or inhibition. The most common neurodegenerative disorders are Alzheimer’s disease, Parkinson’s disease, Amyotrophic Lateral Sclerosis and Huntington’s disease. The molecular features of these disorders have been extensively researched and various unique neurotherapeutic interventions have been developed. However, there is an enormous coercion to integrate the existing knowledge in order to intensify the reliability with which neurodegenerative disorders can be diagnosed and treated. The objective of this review article is therefore to assimilate these disorders’ in terms of their neuropathology, neurogenetics, etiology, trends in pharmacological treatment, clinical management, and the use of innovative neurotherapeutic interventions.

IJMS, Vol. 10, Pages 1872-1895: Hereditary and Sporadic Forms of Aβ-Cerebrovascular Amyloidosis and Relevant Transgenic Mouse Models

Samir Kumar-Singh — Thu, 23 Apr 2009 00:00:00 CEST

Cerebral amyloid angiopathy (CAA) refers to the specific deposition of amyloid fibrils in the leptomeningeal and cerebral blood vessel walls, often causing secondary vascular degenerative changes. Although many kinds of peptides are known to be deposited as vascular amyloid, amyloid-β (Aβ)-CAA is the most common type associated with normal aging, sporadic CAA, Alzheimer’s disease (AD) and Down’s syndrome. Moreover, Aβ-CAA is also associated with rare hereditary cerebrovascular amyloidosis due to mutations within the Aβ domain of the amyloid precursor protein (APP) such as Dutch and Flemish APP mutations. Genetics and clinicopathological studies on these familial diseases as well as sporadic conditions have already shown that CAA not only causes haemorrhagic and ischemic strokes, but also leads to progressive dementia. Transgenic mouse models based on familial AD mutations have also successfully reproduced many of the features found in human disease, providing us with important insights into the pathogenesis of CAA. Importantly, such studies have pointed out that specific vastopic Aβ variants or an unaltered Aβ42/Aβ40 ratio favor vascular Aβ deposition over parenchymal plaques, but higher than critical levels of Aβ40 are also observed to be anti-amyloidogenic. These data would be important in the development of therapies targeting amyloid in vessels.

IJMS, Vol. 10, Pages 1824-1837: In Vitro Cytochrome P450 Formation of a Mono-Hydroxylated Metabolite of Zearalenone Exhibiting Estrogenic Activities: Possible Occurrence of This Metabolite in Vivo

Frederique Bravin — Tue, 21 Apr 2009 00:00:00 CEST

The mycoestrogen zearalenone (ZEN), as well as its reduced metabolites, which belong to the endocrine disruptor bio-molecule family, are substrates for various enzymes involved in steroid metabolism. In addition to its reduction by the steroid dehydrogenase pathway, ZEN also interacts with hepatic detoxification enzymes, which convert it into hydroxylated metabolites (OH-ZEN). Due to their structures to that of estradiol, ZEN and its derived metabolites bind to the estrogen receptors and are involved in endocrinal perturbations and are possibly associated with estrogen-dependent cancers. The primary aim of this present study was to identify the enzymatic cytochrome P450 isoforms responsible for the formation of the most abundant OH-ZEN. We thus studied its in vitro formation using hepatic microsomes in a range of animal model systems including man. OH-ZEN was also recovered in liver and urine of rats treated orally with ZEN. Finally we compared the activity of ZEN and its active metabolites (α-ZAL and OH-ZEN) on estrogen receptors using HeLa ER-α and ER-β reporter cell lines as reporters. OH-ZEN estrogenic activities were revealed to be limited and not as significant as those of ZEN or α-ZAL.

IJMS, Vol. 10, Pages 1465-1475: Mycotoxin Detection in Human Samples from Patients Exposed to Environmental Molds

Dennis G. Hooper — Wed, 01 Apr 2009 00:00:00 CEST

The goal of this study was to determine if selected mycotoxins (trichothecenes, aflatoxins, and ochratoxins) could be extracted and identified in human tissue and body fluids from patients exposed to toxin producing molds in their environment. Human urine and methanol extracted tissues and sputum were examined. Trichothecenes were tested using competitive ELISA techniques. Aflatoxins B1, B2, G1, and G2, and ochratoxin A were tested by using immunoaffinity columns and fluorometry. Test sensitivity and specificity were determined. Levels of detection for the various mycotoxins varied from 0.2 ppb for trichothecenes, 1.0 ppb for aflatoxins, and 2.0 ppb for ochratoxins. Trichothecene levels varied in urine, sputum, and tissue biopsies (lung, liver, brain) from undetectable (<0.2 ppb) to levels up to 18 ppb. Aflatoxin levels from the same types of tissues varied from 1.0 to 5.0 ppb. Ochratoxins isolated in the same type of tissues varied from 2.0 ppb to > 10.0 ppb. Negative control patients had no detectable mycotoxins in their tissues or fluids. These data show that mycotoxins can be detected in body fluids and human tissue from patients exposed to mycotoxin producing molds in the environment, and demonstrate which human tissues or fluids are the most likely to yield positive results.

IJMS, Vol. 10, Pages 1386-1406: Molecular Pathogenesis of Alzheimer’s Disease: Reductionist versus Expansionist Approaches

Rudy J. Castellani — Thu, 26 Mar 2009 00:00:00 CET

Alzheimer’s disease (AD) is characterized clinically by dementia and pathologically by two hallmark lesions, senile plaques and neurofibrillary tangles. About a quarter century ago these hallmark lesions were purified and their protein constituents identified, precipitating an avalanche of molecular studies as well as substantial optimism about successful therapeutic intervention. In 2009, we now have copious knowledge on the biochemical cascades that produce these proteins, the different modifications and forms in which these proteins exist, and the ability to selectively target these proteins for therapeutic intervention on an experimental basis. At the same time, there has been no discernible alteration in the natural course of AD in humans. While it may be that the complexity of AD will exceed our capacity to make significant treatment progress for decades or more, a paradigm shift from the reductionism that defines amyloid-β and tau hypotheses, to one that more accurately reflects the meaning of neuropathological changes, may be warranted. We and others have demonstrated that AD pathology is a manifestation of cellular adaptation, specifically as a defense against oxidative injury. As such, AD pathology is therefore a host response rather than a manifestation of cytotoxic protein injury, and is unlikely to be a fruitful target for therapeutic intervention. An “expansionist” view of the disease, we believe, with oxidative stress as a pleiotropic and upstream process, more aptly describes the relationship between various and numerous molecular alterations and clinical disease.

IJMS, Vol. 10, Pages 1226-1260: Physiological and Pathological Role of Alpha-synuclein in Parkinson’s Disease Through Iron Mediated Oxidative Stress; The Role of a Putative Iron-responsive Element

David Olivares — Tue, 17 Mar 2009 00:00:00 CET

Parkinson’s disease (PD) is the second most common progressive neurodegenerative disorder after Alzheimer's disease (AD) and represents a large health burden to society. Genetic and oxidative risk factors have been proposed as possible causes, but their relative contribution remains unclear. Dysfunction of alpha-synuclein (α-syn) has been associated with PD due to its increased presence, together with iron, in Lewy bodies. Brain oxidative damage caused by iron may be partly mediated by α-syn oligomerization during PD pathology. Also, α-syn gene dosage can cause familial PD and inhibition of its gene expression by blocking translation via a newly identified Iron Responsive Element-like RNA sequence in its 5’-untranslated region may provide a new PD drug target.

IJMS, Vol. 10, Pages 1609-1627: Neuronal Aneuploidy in Health and Disease:A Cytomic Approach to Understand the Molecular Individuality of Neurons

Thomas Arendt — Sun, 15 Mar 2009 00:00:00 CET

Structural variation in the human genome is likely to be an important mechanism for neuronal diversity and brain disease. A combination of multiple different forms of aneuploid cells due to loss or gain of whole chromosomes giving rise to cellular diversity at the genomic level have been described in neurons of the normal and diseased adult human brain. Here, we describe recent advances in molecular neuropathology based on the combination of slide-based cytometry with molecular biological techniques that will contribute to the understanding of genetic neuronal heterogeneity in the CNS and its potential impact on Alzheimer´s disease and age-related disorders.

IJMS, Vol. 10, Pages 1138-1160: Changes in Metallothionein Level in Rat Hepatic Tissue after Administration of Natural Mouldy Wheat

Anna Vasatkova — Thu, 12 Mar 2009 00:00:00 CET

Mycotoxins are secondary metabolites produced by microfungi that are capable of causing disease and death in humans and other animals. This work was aimed at investigation of influence of mouldy wheat contaminated by pathogenic fungi producing mycotoxins on metallothionein levels in hepatic tissue of rats. The rats were administrating feed mixtures with different contents of vitamins or naturally mouldy wheat for 28 days. It was found that the wheat contained deoxynivalenol (80 ± 5 µg per kg of mouldy wheat), zearalenone (56 ± 3 µg/kg), T2-toxin (20 ± 2 µg/kg) and aflatoxins as a sum of B1, B2, G1 and G2 (3.9 ± 0.2 µg/kg). Rats were fed diets containing 0, 33, 66 and 100% naturally moulded wheat. Control group 0, 33, 66 and 100% contained vitamins according to Nutrient Requirements of Rats (NRC). Other four groups (control group with vitamins, vit33, vit66 and vit100%) were fed on the same levels of mouldy wheat, also vitamins at levels 100% higher than the previous mixtures. We determined weight, feed conversion and performed dissection to observe pathological processes. Changes between control group and experimental groups exposed to influence of mouldy wheat and experimental groups supplemented by higher concentration of vitamins and mouldy wheat were not observed. Livers were sampled and did not demonstrate significant changes in morphology compared to control either. In the following experiments the levels of metallothionein as a marker of oxidative stress was determined. We observed a quite surprising trend in metallothionein levels in animals supplemented with increased concentration of vitamins. Its level enhanced with increasing content of mouldy wheat. It was possible to determine a statistically significant decline (p<0.05) between control group and groups of animals fed with 33, 66 and 100% mouldy wheat. It is likely that some mycotoxins presented in mouldy wheat are able to block the mechanism of metallothionein synthesis.

IJMS, Vol. 10, Pages 1045-1063: Molecular Pathology of Neuro-AIDS (CNS-HIV)

Leslie Crews — Wed, 11 Mar 2009 00:00:00 CET

The cognitive deficits in patients with HIV profoundly affect the quality of life of people living with this disease and have often been linked to the neuro-inflammatory condition known as HIV encephalitis (HIVE). With the advent of more effective anti-retroviral therapies, HIVE has shifted from a sub-acute to a chronic condition. The neurodegenerative process in patients with HIVE is characterized by synaptic and dendritic damage to pyramidal neurons, loss of calbindin-immunoreactive interneurons and myelin loss. The mechanisms leading to neurodegeneration in HIVE might involve a variety of pathways, and several lines of investigation have found that interference with signaling factors mediating neuroprotection might play an important role. These signaling pathways include, among others, the GSK3b, CDK5, ERK, Pyk2, p38 and JNK cascades. Of these, GSK3b has been a primary focus of many previous studies showing that in infected patients, HIV proteins and neurotoxins secreted by immune-activated cells in the brain abnormally activate this pathway, which is otherwise regulated by growth factors such as FGF. Interestingly, modulation of the GSK3b signaling pathway by FGF1 or GSK3b inhibitors (lithium, valproic acid) is protective against HIV neurotoxicity, and several pilot clinical trials have demonstrated cognitive improvements in HIV patients treated with GSK3b inhibitors. In addition to the GSK3b pathway, the CDK5 pathway has recently been implicated as a mediator of neurotoxicity in HIV, and HIV proteins might activate this pathway and subsequently disrupt the diverse processes that CDK5 regulates, including synapse formation and plasticity and neurogenesis. Taken together, the GSK3b and CDK5 signaling pathways are important regulators of neurotoxicity in HIV, and modulation of these factors might have therapeutic potential in the treatment of patients suffering from HIVE. In this context, the subsequent sections will focus on reviewing the involvement of the GSK3b and CDK5 pathways in neurodegeneration in HIV.

IJMS, Vol. 10, Pages 1000-1012: Neuropathology and Therapeutic Intervention in Spinal and Bulbar Muscular Atrophy

Haruhiko Banno — Tue, 10 Mar 2009 00:00:00 CET

Spinal and bulbar muscular atrophy (SBMA) is a hereditary motor neuron disease caused by the expansion of a polyglutamine tract in the androgen receptor (AR). The histopathological finding in SBMA is loss of lower motor neurons in the anterior horn of the spinal cord as well as in the brainstem motor nuclei. Animal studies have revealed that the pathogenesis of SBMA depends on the level of serum testosterone, and that androgen deprivation mitigates neurodegeneration through inhibition of nuclear accumulation of the pathogenic AR. Heat shock proteins, ubiquitin-proteasome system and transcriptional regulation are also potential targets of therapy development for SBMA.

IJMS, Vol. 10, Pages 976-999: Molecular Pathology of Human Prion Diseases

Gabor G. Kovacs — Mon, 09 Mar 2009 00:00:00 CET

Prion diseases are fatal neurodegenerative conditions in humans and animals. In this review, we summarize the molecular background of phenotypic variability, relation of prion protein (PrP) to other proteins associated with neurodegenerative diseases, and pathogenesis of neuronal vulnerability. PrP exists in different forms that may be present in both diseased and non-diseased brain, however, abundant disease-associated PrP together with tissue pathology characterizes prion diseases and associates with transmissibility. Prion diseases have different etiological background with distinct pathogenesis and phenotype. Mutations of the prion protein gene are associated with genetic forms. The codon 129 polymorphism in combination with the Western blot pattern of PrP after proteinase K digestion serves as a basis for molecular subtyping of sporadic Creutzfeldt-Jakob disease. Tissue damage may result from several parallel, interacting or subsequent pathways that involve cellular systems associated with synapses, protein processing, oxidative stress, autophagy, and apoptosis.

IJMS, Vol. 10, Pages 954-963: Improvement of Morphine-Mediated Analgesia by Inhibition of β-Arrestin 2 Expression in Mice Periaqueductal Gray Matter

Yuting Li — Thu, 05 Mar 2009 00:00:00 CET

Morphine is a well-known μ-opioid receptor (MOR) agonist and an efficient analgesic, but its long-term use inevitably leads to drug addiction and tolerance. Here, we show that specific inhibition of β-arrestin2 with its siRNA lentivirus microinjected in mice periaqueductal gray matter (PAG) significantly improved both acute and chronic morphine analgesia and delayed the tolerance in the hotplate test. The specific effect of β-arrestin2 was proven by overexpression or knockdown of its homology β-arrestin1 in PAG, which showed no significant effects on morphine analgesia. These findings suggest that specific siRNA targeting β-arrestin2 may constitute a new approach to morphine therapy and other MOR agonist-mediated analgesia and tolerance.

IJMS, Vol. 10, Pages 724-745: Molecular Pathology of Lewy Body Diseases

Katrin Beyer — Thu, 26 Feb 2009 00:00:00 CET

Lewy body diseases are characterized by the presence of Lewy bodies, alpha-synuclein(AS)-positive inclusions in the brain. Since their main component is conformationally modified AS, aggregation of the latter is thought to be a key pathogenic event in these diseases. The analysis of inclusion body constituents gives additional information about pathways also involved in the pathology of synucleinopathies. Widespread mitochondrial dysfunction is very closely related to disease development. The impairment of protein degradation pathways, including both the ubiquitin-proteasome system and the autophagy-lysosomepathway also play an important role during the development of Lewy body diseases. Finally, differential expression changes of isoforms corresponding to genes primarily involved in Lewy body formation point to alternative splicing as another important mechanism in the development of Parkinson’s disease, as well as dementia with Lewy bodies. The present paper attempts to give an overview of recent molecular findings related to the pathogenesis of Lewy body diseases.

IJMS, Vol. 10, Pages 492-506: Expression of Mipu1 in Response to Myocardial Infarction in Rats

Guiliang Wang — Tue, 03 Feb 2009 00:00:00 CET

Myocardial ischemic preconditioning up-regulated protein 1 (Mipu1) was cloned in our laboratory. Male Wistar rats were subjected to left anterior coronary artery ligation and sham-operation and sacrificed at 1 h, 3 h, 6 h, 12 h, 24 h, 3 d or 5 d after ligation. Expression of Mipu1 mRNA and protein were assessed by Northern blotting, real-time quantitative RT-PCR, In Situ hybridization and Western blotting. Expression of Mipu1 was up-regulated at 3 h and lasted to 12 h with a peak at 6 h. Mipu1 mRNA and protein signals express in the endothelium and myocardium in normal and infarcted heart, mainly in infarcted zone. Fluorescent immunocytochemistry showed that Mipu1 protein was localized to the nuclei of H9c2 myogenic cells and was upregulated after the cells being exposed to H2O2. These observations indicates that Mipu1 may play a role in maintaining vascular homeostasis and protecting the myogenic cells from being injured by ischemia-reperfusion or oxidation stress.

IJMS, Vol. 10, Pages 407-440: Drosophila melanogaster as a Model Organism of Brain Diseases

Astrid Jeibmann — Mon, 02 Feb 2009 00:00:00 CET

Drosophila melanogaster has been utilized to model human brain diseases. In most of these invertebrate transgenic models, some aspects of human disease are reproduced. Although investigation of rodent models has been of significant impact, invertebrate models offer a wide variety of experimental tools that can potentially address some of the outstanding questions underlying neurological disease. This review considers what has been gleaned from invertebrate models of neurodegenerative diseases, including Alzheimer’s disease, Parkinson’s disease, metabolic diseases such as Leigh disease, Niemann-Pick disease and ceroid lipofuscinoses, tumor syndromes such as neurofibromatosis and tuberous sclerosis, epilepsy as well as CNS injury. It is to be expected that genetic tools in Drosophila will reveal new pathways and interactions, which hopefully will result in molecular based therapy approaches.

IJMS, Vol. 10, Pages 395-406: Investigation on the Protective Effect of α-Mannan against the DNA Damage Induced by Aflatoxin B1 in Mouse Hepatocytes

Eduardo Madrigal-Santillán — Sun, 01 Feb 2009 00:00:00 CET

Aflatoxin B1 is a contaminant of agricultural and dairy products that can be related to mutagenic and carcinogenic effects. In this report we explore the capacity of α-mannan (Man) to reduce the DNA damage induced by AFB1 in mouse hepatocytes. For this purpose we applied the comet assay to groups of animals which were first administered Man (100, 400 and 700 mg/kg, respectively) and 20 min later 1.0 mg/kg of AFB1. Liver cells were obtained at 4, 10, and 16 h after the chemical administration and examined. The results showed no protection of the damage induced by AFB1 with the low dose of the polysaccharide, but they did reveal antigenotoxic activity exerted by the two high doses. In addition, we induced a co-crystallization between both compounds, determined their fusion points and analyzed the molecules by UV spectroscopy. The obtained data suggested the formation of a supramolecular complex between AFB1 and Man.

IJMS, Vol. 10, Pages 366-384: The Importance of Brain Banks for Molecular Neuropathological Research: The New South Wales Tissue Resource Centre Experience

Irina Dedova — Fri, 23 Jan 2009 00:00:00 CET

New developments in molecular neuropathology have evoked increased demands for postmortem human brain tissue. The New South Wales Tissue Resource Centre (TRC) at The University of Sydney has grown from a small tissue collection into one of the leading international brain banking facilities, which operates with best practice and quality control protocols. The focus of this tissue collection is on schizophrenia and allied disorders, alcohol use disorders and controls. This review highlights changes in TRC operational procedures dictated by modern neuroscience, and provides examples of applications of modern molecular techniques to study the neuropathogenesis of many different brain disorders.

IJMS, Vol. 10, Pages 232-246: Molecular Neuropathology of TDP-43 Proteinopathies

Manuela Neumann — Fri, 09 Jan 2009 00:00:00 CET

The identification of TDP-43 as the major component of the pathologic inclusions in most forms of sporadic and familial frontotemporal lobar degeneration with ubiquitin-positive inclusions (FTLD-U) and amyotrophic lateral sclerosis (ALS) resolved a long-standing enigma concerning the nature of the ubiquitinated disease protein under these conditions. Anti-TDP-43 immunohistochemistry and the recent development of novel tools, such as phosphorylation-specific TDP-43 antibodies, have increased our knowledge about the spectrum of pathological changes associated with FTLD-U and ALS and moreover, facilitated the neuropathological routine diagnosis of these conditions. This review summarizes the recent advances in our understanding on the molecular neuropathology and pathobiology of TDP-43 in FTLD and ALS.

IJMS, Vol. 10, Pages 184-212: Molecular Neuropathology of Gliomas

Markus J. Riemenschneider — Wed, 07 Jan 2009 00:00:00 CET

Gliomas are the most common primary human brain tumors. They comprise a heterogeneous group of benign and malignant neoplasms that are histologically classified according to the World Health Organization (WHO) classification of tumors of the nervous system. Over the past 20 years the cytogenetic and molecular genetic alterations associated with glioma formation and progression have been intensely studied and genetic profiles as additional aids to the definition of brain tumors have been incorporated in the WHO classification. In fact, first steps have been undertaken in supplementing classical histopathological diagnosis by the use of molecular tests, such as MGMT promoter hypermethylation in glioblastomas or detection of losses of chromosome arms 1p and 19q in oligodendroglial tumors. The tremendous progress that has been made in the use of array-based profiling techniques will likely contribute to a further molecular refinement of glioma classification and lead to the identification of glioma core pathways that can be specifically targeted by more individualized glioma therapies.

IJMS, Vol. 10, Pages 174-183: Determination of Aflatoxins in Peanut Products in the Northeast Region of São Paulo, Brazil

Carlos A. F. Oliveira — Tue, 06 Jan 2009 00:00:00 CET

The aim of the present study was to determine aflatoxin levels in peanut products traded in the Northeast region of São Paulo, Brazil. To this end, 240 samples of peanut products traded in the cities of Araras, Leme, Pirassununga and Porto Ferreira were collected from June 2006 to May 2007. The samples were analyzed for aflatoxins (AF) B1, B2, G1 and G2 by high performance liquid chromatography. Results showed 44.2% samples positive for AF at levels of 0.5 to 103.8 mg·kg-1. Nine of the positive samples (3.7% of the analysed samples) had total aflatoxin concentrations (B1+B2+G1+G2) higher than the limit established by Brazilian regulations (20 mg·kg-1). Based on the above data, the probable mean daily intake (PDIM) of aflatoxins from peanut products in the Northeast region of São Paulo was estimated to be 0.23 ng kg b.w. day-1. Although this PDIM value was relatively low, results indicate that aflatoxin contamination of peanut products may be a public health concern in Brazil, when considering the potential exposure of highly susceptible consumers. For example, it should be emphasized that children are potentially exposed to aflatoxins, since they consume large quantities of peanut candies, and these products had the highest number of samples positive for AFB1.

IJMS, Vol. 10, Pages 147-173: Trichothecenes in Cereal Grains

Nora A. Foroud — Tue, 06 Jan 2009 00:00:00 CET

Trichothecenes are sesquiterpenoid mycotoxins associated with fusarium head blight (FHB) of cereals, with worldwide economic and health impacts. While various management strategies have been proposed to reduce the mycotoxin risk, breeding towards FHB-resistance appears to be the most effective means to manage the disease, and reduce trichothecene contamination of cereal-based food products. This review provides a brief summary of the trichothecene synthesis in Fusarium species, their toxicity in plants and humans, followed by the current methods of screening and breeding for resistance to FHB and trichothecene accumulation.

IJMS, Vol. 10, Pages 62-115: An Overview of Conventional and Emerging Analytical Methods for the Determination of Mycotoxins

Irena Kralj Cigić — Fri, 02 Jan 2009 00:00:00 CET

Mycotoxins are a group of compounds produced by various fungi and excreted into the matrices on which they grow, often food intended for human consumption or animal feed. The high toxicity and carcinogenicity of these compounds and their ability to cause various pathological conditions has led to widespread screening of foods and feeds potentially polluted with them. Maximum permissible levels in different matrices have also been established for some toxins. As these are quite low, analytical methods for determination of mycotoxins have to be both sensitive and specific. In addition, an appropriate sample preparation and pre-concentration method is needed to isolate analytes from rather complicated samples. In this article, an overview of methods for analysis and sample preparation published in the last ten years is given for the most often encountered mycotoxins in different samples, mainly in food. Special emphasis is on liquid chromatography with fluorescence and mass spectrometric detection, while in the field of sample preparation various solid-phase extraction approaches are discussed. However, an overview of other analytical and sample preparation methods less often used is also given. Finally, different matrices where mycotoxins have to be determined are discussed with the emphasis on their specific characteristics important for the analysis (human food and beverages, animal feed, biological samples, environmental samples). Various issues important for accurate qualitative and quantitative analyses are critically discussed: sampling and choice of representative sample, sample preparation and possible bias associated with it, specificity of the analytical method and critical evaluation of results.

IJMS, Vol. 10, Pages 37-49: Developmental Toxicity of Ochratoxin A in Rat Embryo Midbrain Micromass Cultures

Iwona Wilk–Zasadna — Sat, 27 Dec 2008 00:00:00 CET

Embryonic midbrain micromass cultures were exposed for five days to ochratoxin A (OTA) at seven concentrations (ranging from 0.16 to 10 ?g/mL). Cell viability was assessed in neutral red uptake test (NRU), and differentiation - by immunoenzymatic determination of structural proteins (?III-tubulin, MAP2, GFAP) expression level as well as by computer image analysis. Dose dependent decrease in cell number and differentiation was observed. Concentration-response curves were analysed and the mean inhibition concentrations (?g/mL) for cytotoxicity (IC50) and differentiation (ID50) were calculated. There were no significant differences in the sensitivity of neurons in early and late stage of differentiation and astrocytes to the toxic activity of this compound. For all endpoints ID50 value was very low (< 10 ?g/mL) so OTA was classified as a strong teratogen. IC50/ ID50 ratios

IJMS, Vol. 10, Pages 1-17: Impact of Deoxynivalenol on the Intestinal Microflora of Pigs

Yann J. Waché — Sat, 27 Dec 2008 00:00:00 CET

Deoxynivalenol (DON), a mycotoxin produced by some Fusarium species, is a frequent contaminant of cereal. In the present study, 24 weanling piglets received either control feed or feed naturally contaminated with DON (2.8 mg/kg) for four weeks. Consumption of contaminated feed significantly reduced the animal weight gain during the first week of the experiment, but had a moderate effect on cultivable bacteria in the pig intestine. By contrast, changes in the intestinal microflora were observed by Capillary Electrophoresis Single-Stranded Conformation Polymorphism (CE-SSCP) in DON-exposed animals, suggesting an impact of this toxin on the dynamics of intestinal bacteria communities.

IJMS, Vol. 9, Pages 2614-2621: Occurrence of Aflatoxin M1 in Dairy Products in Southern Italy

Maria Teresa Montagna — Fri, 12 Dec 2008 00:00:00 CET

A screening survey of the presence of aflatoxin M1 (AFM1) was carried out on 265 samples of cheese made from cow, buffalo, goat, sheep, sheep-goat milk collected in the Apulia region (Southern Italy). Selected samples included unripened, medium and long-term ripened cheeses. AFM1 was found in 16.6% of the analyzed samples. The highest positive incidence was for medium and long-term ripened cheeses, especially those made from sheep-goat milk, while buffalo cheeses tested consistently negative. Our results show that the level of contamination by AFM1 in dairy products from Apulia Region are lower than in other Italian and European regions. Moreover, it is important to underline that a common European norm concerning the AFM1 threshold limits for dairy products is still lacking.

IJMS, Vol. 9, Pages 2585-2600: Analysis of Mechanisms of T-2 Toxin Toxicity Using Yeast DNA Microarrays

Yumiko Iwahashi — Thu, 11 Dec 2008 00:00:00 CET

T-2 toxin is a mycotoxin that belongs to a group of type A tricothecenes found in agricultural products. The cytotoxicity of T-2 toxin was characterized by analysis of the yeast transcriptome upon challenge with T-2 toxin. Interestingly, T-2 toxin-induced yeast gene expression profiles were found to be similar to profiles obtained following cycloheximide treatment. Moreover, T-2 toxin treatment was found to activate facilitators, gluconeogenesis and cell arrest related genes such as mitogen-activated protein kinase genes (FUS3). T-2 toxin attacks the membrane and as a result the membrane transport system was disturbed. A large number of genes are induced to restore the toxicity caused by T-2 toxin. However, the data did not suggest that DNA damage by alkylation (Mag1, a gene 3-methyl-adenine DNA glycosylase, 0.46-fold down regulated), no induction of DNA repair mechanisms such as recombination (RAD26, RAD52 and etc.) and excision repair (RAD7, RAD14, RAD16, RAD23 and etc.). These results suggested that the toxicity of the T-2 toxin was due to the disturbance of the cell membrane of the yeast cell and that T-2 toxin caused mild mutagenesis.

IJMS, Vol. 9, Pages 2570-2584: Zearalenone and Reproductive Function in Farm Animals

Fiorenza Minervini — Thu, 11 Dec 2008 00:00:00 CET

Farm animals are exposed to zearalenone through the feed because of the widespread occurrence of this mycotoxin in cereals and clinical reproductive disorders due to mycotoxin effects are often reported in farm animal species. This review describes the metabolism, the mechanistic aspects, the clinical reproductive symptoms and the in vitro effects on functional parameters of oocytes and sperm cells induced by zearalenone and its derivatives in farm animals. The studies on in vitro effects allow to understand the action mechanisms of mycotoxins and, sometime, to explain the in vivo symptoms. The impairment of semen quality and female reproductive function induced by zearalenone could be a factor responsible for the reproductive failure in farm animals.

IJMS, Vol. 9, Pages 2543-2558: Species-specific Fungal DNA in Airborne Dust as Surrogate for Occupational Mycotoxin Exposure?

Anne Straumfors Halstensen — Wed, 10 Dec 2008 00:00:00 CET

Possible health risks associated with occupational inhalation of mycotoxin-containing dust remain largely unknown, partly because methods for mycotoxin detection are not sensitive enough for the small dust masses obtained by personal sampling, which is needed for inhalable exposure measurements. Specific and sensitive PCR detection of fungi with mycotoxin-producing potential seem to be a good surrogate for occupational exposure measurements that include all fungal structures independent of morphology and cultivability. Results should, however, be interpreted with caution due to variable correlations with mycotoxin concentrations.

IJMS, Vol. 9, Pages 2505-2514: A Nutritional Approach for the Management of Deoxynivalenol (DON) Toxicity in the Gastrointestinal Tract of Growing Chickens

Wageha Awad — Mon, 08 Dec 2008 00:00:00 CET

It has been shown that DON has negative effects on the active transport of some nutrients in the small intestine of chickens. The plausible interactions between food contaminants and natural components could be high. The present study investigated the effects of DON on the presence or absence of dietary inulin on the electrophysiological response of the gut to glucose. Ussing chamber studies were conducted with isolated jejunal epithelia at the age of 35 days. Electrophysiology of the epithelia was recorded and the changes of the short-circuit current (Isc) were determined. The addition of Dglucose on the luminal side of the isolated mucosa increased (P < 0.05) the Isc in the control group and inulin supplemented group. The oligosaccharides did not increase glucose absorption in young healthy chickens compared with the controls. In the second experiment, after preincubation of tissues with DON, the addition of glucose did not increase the Isc in jejunum and colon in the control group (P > 0.05). However, in the dietary inulin supplemented group in both jejunum and colon, the addition of glucose after preincubation of tissues with DON increased the Isc, suggesting that the dietary inulin supplementation of the broilers regulated and improved the glucose absorption in the presence of DON and kept it at normal levels.

IJMS, Vol. 9, Pages 2495-2504: Development of a Generic PCR Detection of 3-Acetyldeoxynivalenol-, 15-Acetyldeoxynivalenol- and Nivalenol-Chemotypes of Fusarium graminearum Clade

Jian-Hua Wang — Fri, 05 Dec 2008 00:00:00 CET

Fusarium graminearum clade pathogens cause Fusarium head blight (FHB) or scab of wheat and other small cereal grains, producing different kinds of trichothecene mycotoxins that are detrimental to human and domestic animals. Type B trichothecene mycotoxins such as deoxynivalenol, 3-acetyldeoxynivalenol (3-AcDON), 15-acetyldeoxynivalenol (15-AcDON) and nivalenol (NIV) are the principal Fusarium mycotoxins reported in China, as well as in other countries. A genomic polymerase chain reaction (PCR) to predict chemotypes was developed based on the structural gene sequences of Tri13 genes involved in trichothecene mycotoxin biosynthesis pathways. A single pair of primers derived from the Tri13 genes detected a 583 bp fragment from 15-AcDON-chemotypes, a 644 bp fragment from 3-AcDON-chemotypes and an 859 bp fragment from NIV-producing strains. Fusarium strains from China, Nepal, USA and Europe were identified by this method, revealing their mycotoxin chemotypes identical to that obtained by chemical analyses of HPLC or GC/MS and other PCR assays. The mycotoxin chemotype-specific fragments were amplified from a highly variable region located in Tri13 genes with three deletions for 15-AcDON-chemotypes, two deletions for 3-AcDON-chemotypes and no deletion for NIV-producers. This PCR assay generated a single amplicon and thus should be more reliable than other PCR-based assays that showed the absence or presence of a PCR fragment since these assays may generate false-negative results. The results with strains from several different countries as well as from different hosts further indicated that this method should be globally applicable. This is a rapid, reliable and cost-effective method for the identification of type B trichothecene mycotoxin chemotypes in Fusarium species and food safety controls.

IJMS, Vol. 9, Pages 2474-2494: Mycotoxin Detection Plays “Cops and Robbers”: Cyclodextrin Chemosensors as Specialized Police?

Pietro Cozzini — Fri, 05 Dec 2008 00:00:00 CET

As in a cops and robbers play we discover new mycotoxins and metabolites everyday and we are forced to develop new molecules quickly as chemo- or biosensors or to modify existing molecules able to recognize these new hazardous compounds. This will result in an enormous cost saving to agro-food industry through the prevention and reduction of product recalls and reduced treatment costs. Here we present a brief review of the rapid methods used to detect mycotoxins, considering usefulness and limits. Then we propose a new fast, efficient and cheap methodology, based on a combination of computer chemistry aided design and fluorescence, that can help to drive synthesis in a more efficient way.

IJMS, Vol. 9, Pages 2366-2375: Isolation and Identification of a Strain of Aspergillus Tubingensis With Deoxynivalenol Biotransformation Capability

Chenghua He — Thu, 27 Nov 2008 00:00:00 CET

Deoxynivalenol (DON) is one of the most common contaminants of various foodstuffs. A biotransformation system was used in order to lessen the toxicity of DON. A strain of Aspergillus (NJA-1) was isolated from soil and cultured in an inorganic salt medium containing DON. Bt2a/Bt2b primers were used to amplify the β-tubulin gene of NJA-1. Sequence analysis the PCR product and morphology observation indicated that NJA-1 belonged to Aspergillus tubingensis (aerobic fungi). The DNA sequence information of the PCR product was deposited in GenBank (accession number DQ9025790). The DNA sequence had 99% similarity to the Aspergillus tubingensis accession number AY820009. An unknown compound in NJA-1 showed the ability to convert DON into another product. The molecular weight of the bioconversion product was 18.1 D (H2O) larger than that of DON. The analysis showed that DON could be hydrolyzed by NJA-1. The mean DON biotransformation rate was 94.4% after two weeks of cultivation. The finding presents a new method for DON biotransformation.

IJMS, Vol. 9, Pages 2357-2365: Growth and Mycotoxin Production by Chaetomium globosum Is Favored in a Neutral pH

Matthew R. Fogle — Wed, 26 Nov 2008 00:00:00 CET

Chaetomium globosum is frequently isolated in water-damaged buildings and produces two mycotoxins called chaetoglobosins A and C when cultured on building material. In this study, the influence of ambient pH on the growth of C. globosum was examined on an artificial medium. This fungus was capable of growth on potato dextrose agar ranging in pH from 4.3 to 9.4 with optimal growth and chaetoglobosin C production occurring at a neutral pH. In addition, our results show that sporulation is favored in an acidic environment.

IJMS, Vol. 9, Pages 2306-2321: Upscaled CTAB-Based DNA Extraction and Real-Time PCR Assays for Fusarium culmorum and F. graminearum DNA in Plant Material with Reduced Sampling Error

Christoph Brandfass — Tue, 25 Nov 2008 00:00:00 CET

Fusarium graminearum Schwabe (Gibberella zeae Schwein. Petch.) and F. culmorum W.G. Smith are major mycotoxin producers in small-grain cereals afflicted with Fusarium head blight (FHB). Real-time PCR (qPCR) is the method of choice for species-specific, quantitative estimation of fungal biomass in plant tissue. We demonstrated that increasing the amount of plant material used for DNA extraction to 0.5-1.0 g considerably reduced sampling error and improved the reproducibility of DNA yield. The costs of DNA extraction at different scales and with different methods (commercial kits versus cetyltrimethylammonium bromide-based protocol) and qPCR systems (doubly labeled hybridization probes versus SYBR Green) were compared. A cost-effective protocol for the quantification of F. graminearum and F. culmorum DNA in wheat grain and maize stalk debris based on DNA extraction from 0.5-1.0 g material and real-time PCR with SYBR Green fluorescence detection was developed.

IJMS, Vol. 9, Pages 2205-2216: Intestinal Structure and Function of Broiler Chickens on Diets Supplemented with a Synbiotic Containing Enterococcus faecium and Oligosaccharides

Wageha Awad — Wed, 12 Nov 2008 00:00:00 CET

A feeding trial was conducted on broiler chickens to study the effects of the synbiotic BIOMIN IMBO [a combination of Enterococcus faecium, a prebiotic (derived from chicory) and immune modulating substances (derived from sea algae)], with a dose of 1 kg/ton of the starter diets and 0.5 kg/ton of the grower diets on the intestinal morphometry and nutrient absorption. The general performance was improved (P < 0.05) by the dietary inclusion of synbiotic compared with the controls. Furthermore, the addition of synbiotic increased (P < 0.001) the villus height/crypt depth ratio and villus height in ileum. However, the ileal crypt depth was decreased by dietary supplementation of synbiotic compared with control. The addition of glucose in Ussing chamber produced a significant increase (P ≤ 0.001) in short-circuit current (Isc) in jejunum and colon relative to the basal values in both synbiotic and control groups. However, in jejunum the percentage of Isc increase after glucose addition was higher for synbiotic group (333 %) than control group (45 %). In conclusion, dietary inclusion of synbiotic BIOMIN IMBO increased the growth performance and improved intestinal morphology and nutrient absorption.

IJMS, Vol. 9, Pages 2169-2183: Survey of Slaughtered Pigs for Occurrence of Ochratoxin A and Porcine Nephropathy in Serbia

Dragan Milićević — Fri, 07 Nov 2008 00:00:00 CET

Samples of blood, kidney and liver were randomly selected from slaughtered pigs (n=90) and analyzed for ochratoxin A by HPLC. In addition, in order to obtain information on the occurrence of nephropathy, histological examinations were carried out. Of the 90 liver samples, 26.6% contained OTA in the range of 0.22-14.5 ng/g. The incidence of OTA in serum and kidney were very similar (31%, 33.3%), with a maximum concentration of 220.8 ng/mL, and 52.5 ng/g, respectively. Histopathological examination of kidneys confirmed tubulopathies with edema and cell vacuolization. In addition, hemorrhages and necrosis of proximal kidney tubules’ cells were found.

IJMS, Vol. 9, Pages 2146-2158: T-2 Toxin-induced Toxicity in Pregnant Mice and Rats

Kunio Doi — Wed, 05 Nov 2008 00:00:00 CET

T-2 toxin is a cytotoxic secondary fungal metabolite that belongs to the trichothecene mycotoxin family. This mycotoxin is a well known inhibitor of protein synthesis through its high binding affinity to peptidyl transferase, which is an integral part of the ribosomal 60s subunit, and it also inhibits the synthesis of DNA and RNA, probably secondary to the inhibition of protein synthesis. In addition, T-2 toxin is said to induce apoptosis in many types of cells bearing high proliferating activity. T-2 toxin readily passes the placenta and is distributed to embryo/fetal tissues, which include many component cells bearing high proliferating activity. This paper reviews the reported data related to T-2 toxin-induced maternal and fetal toxicities in pregnant mice and rats. The mechanisms of T-2 toxin-induced apoptosis in maternal and fetal tissues are also discussed in this paper.

IJMS, Vol. 9, Pages 2127-2145: Effects of Milling and Cooking Processes on the Deoxynivalenol Content in Wheat

Masayo Kushiro — Wed, 05 Nov 2008 00:00:00 CET

Deoxynivalenol (DON, vomitoxin) is a natural-occuring mycotoxin mainly produced by Fusarium graminearum, a food-borne fungi widely distributed in crops and it is one of the most important mycotoxins in wheat and wheat-based foods and feeds. DON affects animal and human health causing diarrhea, vomiting, gastro-intestinal inflammation, and immunomodulation. Since the rate of the occurrence of DON in wheat is high, effective procedures to remove or eliminate DON from food products is essential to minimize exposures in those who consume large amounts of wheat. Cleaning prior to milling reduced to some extent the concentration of DON in final products. Since DON is distributed throughout the kernels, with higher content in the outer skin, milling is also effective in reducing the DON levels of wheat-based foods if bran and shorts are removed before thermal cooking. DON is water-soluble and cooking with larger amounts of water lowers DON content in products such as spaghetti and noodles. During baking or heating, DON is partially degraded to DON-related chemicals, whose toxicological effects are not studied well. This paper reviews the researches on the effects of milling and cooking on the DON level and discusses the perspectives of further studies.

IJMS, Vol. 9, Pages 2114-2126: Occurrence of Deoxynivalenol in Maize and Wheat in Serbia

Igor Jajić — Tue, 04 Nov 2008 00:00:00 CET

A total of 226 samples of maize and 59 of wheat from the 2004−2007 harvests were investigated for the presence and concentration of deoxynivalenol (DON). Samples of the 2004 harvest were analyzed after their storing for one year in barns, while those of the 2005−2007 harvest were taken directly off fields immediately after the harvest. The samples were analyzed by liquid chromatography on an ODS Hypersil column with DAD detector and ELISA methods. The average incidence rate of DON in maize from the 2004 harvest was 50% (concentration range 0.042−2.460 mg/kg, average value 0.536 mg/kg), while for those of the 2005−2007 harvest it was 32.4% (concentration range 0.027−2.210 mg/kg, average value 0.223 mg/kg). In the case of wheat incidence rate of DON for 2004 harvest was 50.0% (concentration range 0.630−1.840 mg/kg, average value 1.235 mg/kg), while for those of the 2005−2007 harvest it was 34.5% (concentration range 0.057−0.423 mg/kg, average value 0.190 mg/kg). Concentrations in two samples of maize and one of wheat (one sample of each cereal being of the 2004 harvest) were above the maximum level adopted by the European Commission. The results obtained were analyzed as a function of climatic conditions and compared with those of the neighboring countries where the relevant data existed.

IJMS, Vol. 9, Pages 2091-2104: Terminal Continuation (TC) RNA Amplification Enables Expression Profiling Using Minute RNA Input Obtained from Mouse Brain

Melissa J. Alldred — Fri, 31 Oct 2008 00:00:00 CET

A novel methodology named terminal continuation (TC) RNA amplification has been developed to amplify RNA from minute amounts of starting material. Utility of the TC RNA amplification method is demonstrated with two new modifications including obviating the need for second strand synthesis, and purifying the amplification template using column filtration prior to in vitro transcription (IVT). Using four low concentrations of RNA extracted from mouse brain (1, 10, 25 and 50 ng), one round TC RNA amplification was compared to one round amplified antisense RNA (aRNA) in conjunction with column filtration and drop dialysis purification. The TC RNA amplification without second strand synthesis performed extremely well on customdesigned cDNA array platforms, and column filtration was found to provide higher positive detection of individual clones when hybridization signal intensity was subtracted from corresponding negative control hybridization signal levels. Results indicate that TC RNA amplification without second strand synthesis, in conjunction with column filtration, is an excellent method for RNA amplification from extremely small amounts of input RNA from mouse brain and postmortem human brain, and is compatible with microaspiration strategies and subsequent microarray analysis.

IJMS, Vol. 9, Pages 2062-2090: Fumonisins, Trichothecenes and Zearalenone in Cereals

Selma Yazar — Fri, 31 Oct 2008 00:00:00 CET

Fumonisins are phytotoxic mycotoxins which are synthesized by various species of the fungal genus Fusarium such as Fusarium verticillioides (Sacc.) Nirenberg (ex F.moniliforme Sheldon) and Fusarium proliferatum. The trichothecene (TC) mycotoxins are secondary metabolites produce by species that belong to several fungal genera, especially Fusarium, Stachybotrys, Trichothecium, Trichoderma, Memnoniella and Myrothecium. Fusarium mycotoxins are widely dispersed in cereals and their products. Zearalenone (ZEA) is an estrogenic compound produced by Fusarium spp. such as F. graminearum and F. culmorum. Fumonisins, the TCs and ZEA are hazardous for human and animal health. Contamination with TCs causes a number of illnesses in human and animal such as decrease in food consumption (anorexia), depression or inhibition on immune system function and haematoxicity. The purpose of this paper is to give a review of the papers published on the field of fumonisin, TC and ZEA mycotoxins in cereals consumed in the world.

IJMS, Vol. 9, Pages 2027-2043: Aspergillus parasiticus crzA, Which Encodes Calcineurin Response Zinc-Finger Protein, Is Required for Aflatoxin Production under Calcium Stress

Perng-Kuang Chang — Wed, 29 Oct 2008 00:00:00 CET

Two morphologically different Aspergillus parasiticus strains, one producing aflatoxins, abundant conidia but few sclerotia (BN9) and the other producing O-methylsterimatocystin (OMST), copious sclerotia but a low number of conidia (RH), were used to assess the role of crzA which encodes a putative calcium-signaling pathway regulatory protein. Under standard culture conditions, BN9ΔcrzA mutants conidiated normally but decreased slightly in radial growth, regardless of illumination conditions. RHΔcrzA mutants produced only conidia under light and showed decreased conidiation and delayed sclerotial formation in the dark. Regulation of conidiation of both A. parasiticus strains by light was independent of crzA. Increased concentrations of lithium, sodium, and potassium impaired conidiation and sclerotial formation of the RHΔcrzA mutants but they did not affect conidiation of the BN9ΔcrzA mutants. Vegetative growth and asexual development of both ΔcrzA mutants were hypersensitive to increased calcium concentrations. Calcium supplementation (10 mM) resulted in 3-fold and 2-fold decreases in the relative expression of the endoplasmic reticulum calcium ATPase 2 gene in the BN9 and RH parental strains, respectively, but changes in both ΔcrzA mutants were less significant. Compared to the parental strains, the ΔcrzA mutants barely produced aflatoxins or OMST after the calcium supplementation. The relative expression levels of aflatoxin biosynthesis genes, nor1, ver1, and omtA, in both ΔcrzA mutants were decreased significantly, but the decreases in the parental strains were at much lower extents. CrzA is required for growth and development and for aflatoxin biosynthesis under calcium stress conditions.