A. Bernadotte

Peptide Regulation of Gene Expression and Protein Synthesis in Bronchial Epithelium

IntroductionSome studies have shown that peptides have high treatment potential due to their biological activity, harmlessness, and tissue-specific action. Tetrapeptide Ala-Asp-Glu-Leu (ADEL) was effective on models of acute bacterial lung inflammation, fibrosis, and toxic lung damage in several studies.MethodsWe measured Ki67, Mcl-1, p53, CD79, and NOS-3 protein levels in the 1st, 7th, and 14th passages of bronchoepithelial human embryonic cell cultures. Gene expression of NKX2-1, SCGB1A1, SCGB3A2, FOXA1, FOXA2, MUC4, MUC5AC, and SFTPA1 was measured by real-time polymerase chain reaction. Using the methods of spectrophotometry, viscometry, and circular dichroism, we studied the ADEL–DNA interaction in vitro.ResultsPeptide ADEL regulates the levels of Ki67, Mcl-1, p53, CD79, and NOS-3 proteins in cell cultures of human bronchial epithelium in various passages. The strongest activating effect of peptide ADEL on bronchial epithelial cell proliferation through Ki67 and Mcl-1 was observed in “old” cell cultures. ADEL regulates the expression of genes involved in bronchial epithelium differentiation: NKX2-1, SCGB1A1, SCGB3A2, FOXA1, and FOXA2. ADEL also activates several genes, which reduced expression correlated with pathological lung development: MUC4, MUC5AC, and SFTPA1. Spectrophotometry, viscometry, and circular dichroism showed ADEL–DNA interaction, with a binding region in the major groove (N7 guanine).ConclusionsADEL can bind to specific DNA regions and regulate gene expression and synthesis of proteins involved in the differentiation and maintenance of functional activity of the bronchial epithelium. Through activation of some specific gene expression, peptide ADEL may protect the bronchial epithelium from pulmonary pathology. ADEL also may have a geroprotective effect on bronchial tissue.

In silico identification and biochemical characterization of the human dicarboxylate clamp TPR protein interaction network

Dicarboxylate clamp tetratricopeptide repeat (dcTPR) motif‐containing proteins are well‐known partners of the heat shock protein (Hsp) 70 and Hsp90 molecular chaperones. Together, they facilitate a variety of intracellular processes, including protein folding and maturation, protein targeting, and protein degradation. An extreme C‐terminal sequence, the EEVD motif, is identical in Hsp70 and Hsp90, and is indispensable for their interaction with dcTPR proteins. However, almost no information is available on the existence of other potential dcTPR‐interacting proteins. We searched the human protein database for proteins with C‐terminal sequences similar to that of Hsp70/Hsp90 to identify potential partners of dcTPR proteins. The search identified 112 proteins containing a Hsp70/Hsp90‐like signature at their C termini. Gene Ontology enrichment analysis of identified proteins revealed enrichment of distinct protein classes, such as molecular chaperones and proteins of the ubiquitin–proteasome system, highlighting the possibility of functional specialization of proteins containing a Hsp70/Hsp90‐like signature. We confirmed interactions of selected proteins containing Hsp70/Hsp90‐like C termini with dcTPR proteins both in vitro and in situ. Analysis of interactions of 10‐amino‐acid peptides corresponding to the C termini of identified proteins with dcTPR proteins revealed significant differences in binding strength between various peptides. We propose a hierarchical mode of interaction within the dcTPR protein network. These findings describe a novel dcTPR protein interaction networks and provide a rationale for selective regulation of protein–protein interactions within this network.

Interleukin-1β, interleukin-1 receptor antagonist, interleukin-6, interleukin-10, and tumor necrosis factor-α levels in CSF and serum in relation to the clinical diversity of Parkinson’s disease

Several parameters representing the clinical diversity of Parkinsons disease (PD), including severity, phenotypes, cognitive decline, anxiety and depression were analyzed to examine the link with interleukin-1β (IL-1β), the interleukin-1 receptor antagonist (IL-1RA), IL-6, IL-10, and tumor necrosis factor-α (TNFα) and also to determine the relationship between levels of these factors in serum and cerebrospinal fluid (CSF). Significantly elevated serum IL-1β and IL-6 and reduced IL-1RA levels were found in the PD group. In CSF and serum, inflammatory factors behaved differently, with increased CSF TNFα indicating rapid PD progression, and increased IL-1β in serum. A low level of IL-6 was associated with a longer duration of PD. Anxiety, depression, non-tremor phenotype and late-onset PD correlated with a high serum level of IL-10. The serum TNFα level was lower in PD patients with mild cognitive impairment compared to controls. Serum IL-1β, IL-6 and IL-10 levels correlated with CSF markers.

The Lewy Body Dementias: Dementia with Lewy Bodies and Related Syndromes

The Lewy body dementias comprise disorders characterized by parkinsonism and a dementia syndrome with a predominant attentional, executive and visuospatial component, and with the Lewy body as the core pathological marker. Dementia with Lewy bodies (DLB) and Parkinson’s disease with dementia (PDD) are two conditions that meet this definition and are only differentiated by the duration of parkinsonism prior to dementia onset. The standardized clinical criteria for DLB establish an arbitrary 1-year cutoff between symptoms: if dementia begins at the same time or within 1 year of parkinsonism, the patient is presumed to suffer from DLB; if dementia onset occurs more than 1 year after parkinsonism, PDD is assumed. The third report of the DLB consortium [1] highlighted the unresolved boundary issues between DLB and PDD but maintained this arbitrary cut point. The appropriateness of such an artificial cut point for differentiating two similar clinical syndromes has been debated. Indeed, DLB and PDD share many common characteristics and might represent different clinical manifestations of a common underlying process. Even within the PDD group, different cognitive profiles exist [2, 3], and some genetic alterations cause both PDD and DLB within the same family [4]. Thus, it is reasonable to consider these conditions as a continuum of possible symptoms: different patient profiles can be identified but with considerable overlap between groups. For this reason, progressive focus has shifted to defining the early stages of these diseases, where differences may be more apparent or clinically recognizable. In this chapter, we aim to present what is currently known about DLB and PDD, focusing on clinical aspects of diagnosis, treatment, and care as well as a taste of foreseeable developments in the near future.

Self-assembly of Deinococcus radiodurans supports nanocell scenario of life origin

In many origin-of-life scenarios, first a kit of elements and simple compounds emerges, then a primitive membrane and then a nanocell with a minimal genome is self-assembled, which then proceeds to multiply by copying itself while mutating. Testing this scenario, we selected Deinococcus Radiodurans known for its exceptional self-repair properties as a model system, separated its bacterial lysis into DNA, RNA and protein fractions, while lipids were used for liposome formation. The fractions were sealed in glass tubes individually and in combinations and stored for three weeks. Upon seeding on Petri dishes, the fractions containing liposomes together with nucleic acid and/or proteins gave in total 19 colonies of Deinococcus radiodurans (confirmed by proteomics), while liposome-free fractions as well as liposome-only fractions gave no colonies. The self-assembly of viable cells from essentially dead mixtures validates the lyposome-based origin-of-life scenario.

Influence of donor age on cellular ability to carry out DNA repair via homologous recombination

Cells from old and young donors differ in their abilities for homologous recombination in vitro, which is distinctly visible at the stage of convergence of the homologous chromosomes. We hypothesize that this effect contributes to genome instability and aging. Cells from donors 70 years and older have a limited ability for homologous recombination induced by X-ray irradiation. Interestingly, a repressor of RNA polymerase II, alpha-amanitin, induces homologous recombination when administered at physiologically toxic doses in cells from donors in all age groups, old and young, as well as in cells with genetic defects in the DNA repair system due to mutations in BRCA1 and BRCA2 genes. Furthermore, in BRCA1 and BRCA2 mutant cells, the effects of irradiation and alpha-amanitin damage are not integrated and do not increase the rate of homologous recombination.