Application of "Omics" Technologies for Diagnosis and Pathogenesis of Neurological Infections.

Infections of the human nervous system have substantial morbidity and mortality but also represent among the most challenging of all neurological diseases because of the difficulty in establishing a diagnosis and implementing effective therapies. Neurological infections lead to altered expression levels of a wide range of host- and pathogen-derived biomolecules both within and outside of the nervous system. Quantitative analyses of these biomolecular perturbations have been traditionally performed using "classical" molecular or analytical methods, which evaluate one or few genes or their products at a time. Recent technical developments together with the increasing availability of high-throughput/content methodologies have enabled a more comprehensive overview of these molecular alterations and thus provide new approaches to the diagnosis and/or treatment of this group of disorders. Herein, we will review recent evidence pointing to the capacity of the so-called omics techniques in studying the nervous system infections with an emphasis on genomics, transcriptomics, and proteomics technologies.

Cardiac chronotropic hypo-responsiveness and atrial fibrosis in rats chronically treated with lithium.

Lithium is a widely used mood-stabilizing agent; however, it causes a variety of cardiovascular side effects including sinus node dysfunction. In this study we explored the potential adverse effects of lithium on cardiac chronotropic responsiveness, atrial tissue histology and gene expression in rats that were chronically treated with therapeutic doses of lithium. Male Wistar albino rats were given lithium chloride (2.5 g/kg) orally for 2 or 3 months. Following treatment, the atria were isolated and spontaneously beating rate and chronotropic responsiveness to ß-adrenergic stimulation was evaluated in an organ bath. Development of cardiac fibrosis was examined by histological methods. The expression of atrial Col1a1 (collagen I, alpha 1) and ß-arrestin2 was also assessed using quantitative RT-PCR. Treatment with lithium induced a significant hypo-responsiveness to adrenergic stimulation (P < 0.001) and caused fibrosis in the atrial tissue of treated rats. In addition, the expression of atrial Col1a1 mRNA was significantly increased in atrial tissues of lithium-treated animals, while ß-arrestin2 mRNA expression did not show a significant difference compared with control animals. Altogether, these findings indicate that cardiac chronotropic hypo responsiveness and associated cardiac fibrosis are side effects of chronic lithium treatment. Moreover, it seems that lithium treatment does not influence ß-arrestin2 mRNA expression.

PLP1 Mutations in Patients with Multiple Sclerosis: Identification of a New Mutation and Potential Pathogenicity of the Mutations.

PLP1 is located on the X-chromosome and encodes myelin proteolipid protein (PLP), the most abundant protein in central nervous system myelin. Generally, point mutations in PLP1 result in X-linked dysmyelinating disorders, such as Pelizaeus-Merzbacher disease (PMD) or spastic paraplegia type 2 (SPG2). However, several case studies have identified patients with missense point mutations in PLP1 and clinical symptoms and signs compatible with a diagnosis of multiple sclerosis (MS). To investigate if PLP1 mutations occur relatively frequently in MS, we sequenced the coding regions of PLP1 in 22 female MS patients who had developed disease after the age of 40 and in 42 healthy women, and identified a missense mutation in exon 2 of PLP1 resulting in a Leu30Val mutation in the protein in one of the MS patients. mCherry-tagged plasmids containing wild type or mutant PLP1 sequences of PLP, including two known PMD/SPG2-related mutations as positive controls, were constructed and transfected into Cos-7 cells. In comparison with cells transfected with wild type PLP1, all mutations caused significant accumulation of PLP in the endoplasmic reticulum of the cells and induction of the unfolded protein response-a mechanism that leads to apoptosis of cells expressing mutant proteins. Additionally, in silico analysis of the binding of peptides containing the Leu30Val mutation to the human leukocyte antigen (HLA) molecules carried by the patient harboring this mutation suggested that the mutation could produce several novel immunogenic epitopes in this patient. These results support the idea that mutations in myelin-related genes could contribute to the development of MS in a small proportion of patients.

Enhancement of Cisplatin Nephrotoxicity by Morphine and Its Attenuation by the Opioid Antagonist Naltrexone.

Nephrotoxicity is a major side effect of cisplatin, a widely used chemotherapy agent. Morphine and other opioids are also used extensively in different types of cancer for the clinical management of pain associated with local or metastatic neoplastic lesions. In addition to its analgesic effects, morphine has also been reported to possess potential immunomodulatory and antioxidant properties. Herein, we investigated the effects of morphine in a rat model of cisplatin-induced nephrotoxicity. Following administration of a single dose of cisplatin (5 mg/kg), animals received intraperitoneal injections of morphine (5 mg/kg/day) and/or naltrexone (20 mg/kg/day), an opioid antagonist, for 5 days. Cisplatin-induced nephrotoxicity was detected by a significant increase in plasma urea and creatinine levels in addition to alterations in kidney tissue morphology. Levels of TNF-α and IL-1ß were significantly increased in the renal tissue in cisplatin group. Moreover, glutathione (GSH) concentration and superoxide dismutase activity were significantly reduced in renal tissue in cisplatin group compared with control animals. Treatment with morphine aggravated the deleterious effects of cisplatin at clinical, biochemical and histopathological levels; whereas naltrexone diminished the detrimental effects of morphine in animals receiving morphine and cisplatin. Morphine or naltrexone alone had no effect on the mentioned parameters. Our findings indicate that concomitant treatment with morphine might intensify cisplatin-induced renal damage in rats. These findings suggest that morphine and other opioids should be administered cautiously in patients receiving cisplatin chemotherapy.

Methadone diminishes neuroinflammation and disease severity in EAE through modulating T cell function.

Methadone is known to exert modulatory effects on the immune system. We investigated the potential effects of methadone on infiltration of inflammatory cells into the spinal cord, as well as the proliferative and cytokine responses of T cells in MOG(35-55)-induced experimental autoimmune encephalomyelitis in mice. Methadone significantly suppressed clinical signs of the disease and level of inflammatory cytokines (p<0.05) produced by T cells. Moreover, invasion of inflammatory cells into the spinal cord was significantly decreased by methadone (p<0.05). Our data point to therapeutic effects of methadone and highlight the beneficial role of opioid receptor signaling in the context of autoimmune neuroinflammation.

Performance of pregnant women on folic acid intake.

The cause of neural tube defects (NTDs) is multifactorial and in this case folic acid has an important role. Since the neural tube is closed during 21-28 days of pregnancy, most of women are not informed about their pregnancy at this time, and as a result the golden time of folic acid consumption is missed. The aim of this study was evaluating the performance of pregnant women attending to Tehran Women's Hospital in regard to folic acid intake during pre-conceptional period between 2011 and 2012. This cross-sectional study was conducted in 370 pregnant women attending the prenatal clinic of a hospital affiliate to Tehran University of Medical Sciences between 2011 and 2012. Data were collected through interview using a questionnaire. Although 70% of the pregnancies were planned, but 70.5% of pregnant women had not taken folic acid before conception or in necessary time. There was found a significant relationship between level of education, history of abnormalities in children and the number of abortions and taking folic acid before pregnancy (P=0.005, P=0.000 and P=0.000, respectively).

Pioglitazone potentiates development of morphine-dependence in mice: possible role of NO/cGMP pathway.

Peroxizome proliferator-activated receptor gamma (PPARγ) is highly expressed in the central nervous system where it modulates numerous gene transcriptions. Nitric oxide synthase (NOS) expression could be modified by simulation of PPARγ which in turn activates nitric oxide (NO)/soluble guanylyl-cyclase (sGC)/cyclic guanosine mono phosphate (cGMP) pathway. It is well known that NO/cGMP pathway possesses pivotal role in the development of opioid dependence and this study is aimed to investigate the effect of PPARγ stimulation on opioid dependence in mice as well as human glioblastoma cell line. Pioglitazone potentiated naloxone-induced withdrawal syndrome in morphine dependent mice in vivo. While selective inhibition of PPARγ, neuronal NOS or GC could reverse the pioglitazone-induced potentiation of morphine withdrawal signs; sildenafil, a phosphodiesterase-5 inhibitor amplified its effect. We also showed that nitrite levels in the hippocampus were significantly elevated in pioglitazone-treated morphine dependent mice. In the human glioblastoma (U87) cell line, rendered dependent to morphine, cAMP levels did not show any alteration after chronic pioglitazone administration while cGMP measurement revealed a significant rise. We were unable to show a significant alteration in neuronal NOS mRNA expressions by pioglitazone in mice hippocampus or U87 cells. Our results suggest that pioglitazone has the ability to enhance morphine-dependence and to augment morphine withdrawal signs. The possible pathway underlying this effect is through activation of NO/GC/cGMP pathway.