Perturbing the Ubiquitin Pathway Reveals How Mitosis Is Hijacked to Denucleate and Regulate Cell Proliferation and Differentiation In Vivo

The eye lens presents a unique opportunity to explore roles for specific molecules in cell proliferation, differentiation and development because cells remain in place throughout life and, like red blood cells and keratinocytes, they go through the most extreme differentiation, including removal of nuclei and cessation of protein synthesis. Ubiquitination controls many critical cellular processes, most of which require specific lysines on ubiquitin (Ub). Of the 7 lysines (K) least is known about effects of modification of K6.We replaced K6 with tryptophan (W) because K6 is the most readily modified K and W is the most structurally similar residue to biotin. The backbone of K6W-Ub is indistinguishable from that of Wt-Ub. K6W-Ub is effectively conjugated and deconjugated but the conjugates are not degraded via the ubiquitin proteasome pathways (UPP). Expression of K6W-ubiquitin in the lens and lens cells results in accumulation of intracellular aggregates and also slows cell proliferation and the differentiation program, including expression of lens specific proteins, differentiation of epithelial cells into fibers, achieving proper fiber cell morphology, and removal of nuclei. The latter is critical for transparency, but the mechanism by which cell nuclei are removed has remained an age old enigma. This was also solved by expressing K6W-Ub. p27(kip), a UPP substrate accumulates in lenses which express K6W-Ub. This precludes phosphorylation of nuclear lamin by the mitotic kinase, a prerequisite for disassembly of the nuclear membrane. Thus the nucleus remains intact and DNAseIIβ neither gains entry to the nucleus nor degrades the DNA. These results could not be obtained using chemical proteasome inhibitors that cannot be directed to specific tissues.K6W-Ub provides a novel, genetic means to study functions of the UPP because it can be targeted to specific cells and tissues. A fully functional UPP is required to execute most stages of lens differentiation, specifically removal of cell nuclei. In the absence of a functional UPP, small aggregate prone, cataractous lenses are formed

Translation and validation of the vertigo symptom scale into German: A cultural adaption to a wider German-speaking population

BACKGROUND: Dizziness and comorbid anxiety may cause severe disability of patients with vestibulopathy, but can be addressed effectively with rehabilitation. For an individually adapted treatment, a structured assessment is needed. The Vertigo Symptom Scale (VSS) with two subscales assessing vertigo symptoms (VSS-VER) and associated symptoms (VSS-AA) might be used for this purpose. As there was no validated VSS available in German, the aim of the study was the translation and cross-cultural adaptation in German (VSS-G) and the investigation of its reliability, internal and external validity. METHODS: The VSS was translated into German according to recognized guidelines. Psychometric properties were tested on 52 healthy controls and 202 participants with vestibulopathy. Internal validity and reliability were investigated with factor analysis, Cronbach's α and ICC estimations. Discriminant validity was analysed with the Mann-Whitney-U-Test between patients and controls and the ROC-Curve. Convergent validity was estimated with the correlation with the Hospital Anxiety Subscale (HADS-A), Dizziness Handicap Inventory (DHI) and frequency of dizziness. RESULTS: Internal validity: factor analysis confirmed the structure of two subscales. Reliability: VSS-G: α = 0.904 and ICC (CI) =0.926 (0.826, 0.965). Discriminant validity: VSS-VER differentiate patients and controls ROC (CI) =0.99 (0.98, 1.00). Convergent validity: VSS-G correlates with DHI (r = 0.554) and frequency (T = 0.317). HADS-A correlates with VSS-AA (r = 0.452) but not with VSS-VER (r = 0.186). CONCLUSIONS: The VSS-G showed satisfactory psychometric properties to assess the severity of vertigo or vertigo-related symptoms. The VSS-VER can differentiate between healthy subjects and patients with vestibular disorders. The VSS-AA showed some screening properties with high sensitivity for patients with abnormal anxiety