Analysis of gene expression in human bullous keratopathy corneas containing limiting amounts of RNA

PURPOSE: To validate the use of polymerase chain reaction (PCR)-amplified full-length cDNA as a substitute for mRNA in nucleic acid array and gene expression analysis. METHODS: Total RNA was isolated from age-matched normal autopsy corneas and pseudophakic bullous keratopathy (PBK) corneas. Full-length cDNA was generated and PCR amplified using the Smart cDNA synthesis technology. Southern blot analysis of this cDNA was compared with Northern blot analysis of the RNA. Amplified cDNA was used to probe a commercial gene array. By immunohistochemistry, the expression pattern of several adhesion molecules represented on the array was assessed. RESULTS: The cDNA produced by the Smart cDNA system gave results very similar to those of northern blot analysis when examined for beta2-microglobulin, Rab geranylgeranyl transferase, and tenascin-C. This cDNA obtained from normal or PBK corneas was labeled and used to probe a 588 gene array (Clontech). Among other differences, beta6 integrin was detected only with the PBK probe, beta-catenin was markedly elevated in PBK, and beta4 integrin appeared to be reduced in PBK. Immunohistochemical patterns of these proteins were consistent with the hybridization signals on the gene array. CONCLUSIONS: Smart cDNA synthesis and nucleic acid arrays were combined and validated for the first time to identify differential gene expression in normal and diseased corneas. These techniques require very little RNA such as that equivalent to a half of a single cornea, which is useful when the amount of tissue is limiting. Altered expression of adhesive proteins beta6 integrin and beta-catenin may be related to the formation of epithelial bullae and microcystic changes in PBK patients.     

Synergism in replication and translation of messenger RNA in a cell-free system.

Combination of the Q beta replicase reaction with the Escherichia coli cell-free translation system markedly enhances replication of a recombinant RQ-DHFR RNA consisting of the dihydrofolate reductase (DHFR) mRNA sequence inserted into RQ135(-1) RNA, an efficient naturally occurring Q beta replicase template. The enhancement is associated with a replication asymmetry previously described for the replication of Q beta phage RNA in vivo; the sense (+)-strands are produced in large excess over the antisense (-)-strands. This, in turn, results in increased synthesis of the functionally active DHFR. These effects are not observed when DHFR mRNAs or RQ135(-1) RNAs are used as templates, if the translation system is not complete, or if it is inhibited by puromycin. The coupled replication-translation of nonviral mRNA recombinants can serve as a useful model for studying the fundamental aspects of virus amplification and can be implemented for large-scale protein synthesis in vitro.     

Neurotransmitter bioamine supply of thymic and lymph node structures during treatment with somatotropic hormone

The effects of somatotropic hormone on bioamine-containing structures of immunocompetent organs in rats were studied after 1, 3, 7, and 14 days of treatment by fluorescent histochemical methods and microspectrofluorometry. The effect of somatotropic hormone on immune processes can be mediated by changes in the neurotransmitter content in the thymus and lymph nodes. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 129, No. 5, pp. 591–593, May, 2000     

Neuropathic Pain in Multiple Sclerosis Patients

Neuroscience and Behavioral Physiology - Among a multitude of pain syndromes (PS) of diverse locations and types of pathogenesis encountered in multiple sclerosis (MS) patients, most research...     

Vaccination and Multiple Sclerosis – Current Situation

Neuroscience and Behavioral Physiology - Active immunization of patients with autoimmune diseases is a current challenge. Vaccination of patients with multiple sclerosis (MS) has been shown not to...     

State of the cervical section of the spinal cord in patients with remitting multiple sclerosis during immunomodulatory treatment

MRI scans were obtained of the cervical section of the spinal cords of 30 patients with remitting multiple sclerosis. During the study period, patients received immunomodulatory agents (seven received interferon β-1a, 13 received interferon β-1b, and 10 received glatiramer acetate). Total focus volume in brain matter was assessed before and after treatment, along with the linear size of the spinal cord on sagittal sections at the level of the inferior margin of the body of C2. There was a significant (p = 0.002) reduction in focus volume in the group overall, from 10993 mm³ (8098–13888 mm³, p < 0.05; Me = 9336) to 5630 mm³ (7400–3860 mm³, p < 0.05, Me = 4180). There were also significant decreases in focus volume on the background of treatment with interferon β-1b and glatiramer acetate (p = 0.026 and 0.027, respectively). Significant differences between groups were found in the magnitudes of increases in spinal cord atrophy: H (2, n = 30) = 8.06; p = 0.0178. Patients given glatiramer acetate showed a significantly smaller increase in atrophy as compared with those treated with interferon β (p < 0.02). Translated from Zhurnal Nevrologii i Psikhiatrii imeni S. S. Korsakova, Multiple Sclerosis, Supplement, No. 4, pp. 129–132, 2007.     

Possible mechanisms of the formation of chronic fatigue syndrome in the clinical picture of multiple sclerosis

A frequent manifestation of multiple sclerosis (MS) is chronic fatigue syndrome, which can be defined as a subjective decrease in the level of physical and/or mental energy. Chronic fatigue syndrome can be divided into asthenia (fatigue at rest), pathological fatigability (fatigue on physical loading), and fatigue on the background of deterioration of other symptoms (exacerbation of MS). There are both central and peripheral mechanisms for the formation of fatigue. The combination of fatigue and affective disturbances, especially depression and sleep disorders (insomnia, restless legs syndrome) is common in MS and may provide evidence that they share common mechanisms — decreases in the activity of the serotoninergic and noradrenergic systems. An important component in the formation of chronic fatigue syndrome consists of endocrine and autoimmune factors, the latter having a greater effect on asthenia than on pathological fatigue. Further studies of the pathogenetic mechanisms of the formation of asthenia and pathological fatigue and clarification of their differential diagnostic signs should allow not only a better understanding of the nature of this syndrome, but also better selection of individual treatment. __________ Translated from Zhurnal Nevrologii i Psikhiatrii imeni S. S. Korsakova, Multiple Sclerosis, Supplement, No. 3, pp. 87–91, 2006.