[The use of the Mini-Cog, MMSE, and GPCOG tests in domestic clinical practice for the evaluation of cognitive disorders in elderly and senile patients: the results of a survey of experts' opinions]. / Primenenie v otechestvennoi klinicheskoi praktike testov Mini-Kog, MMSE i GPCOG dlya otsenki kognitivnykh narushenii u patsientov pozhilogo i starcheskogo vozrasta: rezul'taty oprosa mneniya spetsialistov.

OBJECTIVE: To explore the experts' opinion on the feasibility of using Mini-Kog, MMSE and GPCOG tests for screening assessment of cognitive impairment in elderly and senile patients at the primary stage of medical care, as well as to compare different characteristics of these tests according to experts' opinion. MATERIAL AND METHODS: The survey of specialists was carried out on the basis of 6 Medical Centers. Prior to the survey, specialists tested cognitive functions in elderly and senile patients during routine visits undergoing routine admission using Mini-Kog, MMSE and GPCOG tests, as well as interviewed their relatives using the corresponding section of the GPCOG. During the survey, specialists filled out a special physician checklist containing items for indicating socio-demographic information and questions regarding the use of tests for assessing cognitive functions in elderly and senile patients in daily clinical practice. RESULTS: The survey involved 40 specialists from different cities of Russia (mean age±SD - 38.6±14.3 years, 82.5% - women). Elderly and senile patients accounted for the predominant proportion (76-100%) of all followed-up patients. All physicians considered it important to conduct a cognitive assessment in elderly patients and they have to use various cognitive tests in their practice. According to most experts, among the Mini-Kog, MMSE and GPCOG tests, all three tools are informative, convenient and suitable for screening cognitive impairment in geriatric patients. The MMSE test is more informative and convenient. Mini-Kog, compared to the other two tools, is the simplest and most time-efficient tool that is also better perceived by patients. An advantage of the GPCOG is the possibility of participation of informants in the assessment of cognitive functions in older patients. CONCLUSION: The findings could be used in further research focused on improving the approaches for early detection of cognitive impairment in geriatric patients by primary care physicians.

Comparative In Vitro Study of Antiherpetic Activity of Echinochrome A and Product of Its Oxidation Dehydroechinochrome.

The quinoid pigment echinochrome A isolated from the sea urchin Scaphechinus mirabilis, the product of its oxidation dehydroechinochrome, and structurally similar antiviral agent oxolin were tested for their ability to inhibit plaque formation induced by herpes simplex virus type 1 (HSV-1) in Vero cells. The tested compounds showed significant anti-HSV-1 activity, mainly due to their direct effect on viral particles and on virus attachment to cells. The antiviral efficacy of the test compounds increased in the following order: oxolin→echinochrome A→dehydroechinochrome.

Cytoprotective Effect of Echinochrome A in Primary Culture of Pulmonary Fibroblasts from Albino Rats under Conditions of Oxidative Stress.

We studied the effect of echinochrome A on the primary culture of pulmonary fibroblasts under normal conditions and under oxidative stress. Exposure to echinochrome A (100 µM, 6 h) did not affect the production of superoxide radical by fibroblasts, area of their nuclei, and number of nucleoli, but reduced the total area of nucleolar organizer regions against the background of stable proliferative activity of the culture. Two-hour oxidative stress induced by hydrogen peroxide solution (60 µM) increased the generation of superoxide radical, decreased DNA-synthetic activity of fibroblasts, area of fibroblast nuclei, and total area of nucleolar organizer regions. Preliminary administration of echinochrome A significantly attenuated the damaging effect of oxidative stress: the intensity of production of superoxide radicals decreased, DNA-synthetic activity and nucleus area of fibroblasts partially recovered; normalization of the total area of nucleoli was accompanied by an increase in their number.

Activation of anthraquinone biosynthesis in long-cultured callus culture of Rubia cordifolia transformed with the rolA plant oncogene.

The RolA protein belongs to the RolB class of plant T-DNA oncogenes, and shares structural similarity with the papilloma virus E2 DNA-binding domain. It has potentially as an inducer of plant secondary metabolism, although its role in biotechnology has yet to be realised. In this investigation, a Rubia cordifolia callus culture transformed with the rolA plant oncogene for more than 10 years was analysed. Expression of the rolA gene in the callus line was stable during long-term cultivation, and growth parameters were both elevated and stable, exceeding those of the non-transformed control culture. The rolA-transformed calli not only demonstrated remarkably stable growth, but also the ability to increase the yield of anthraquinones (AQs) in long-term cultivation. After ten years of cultivating rolA callus lines, we observed an activation of AQ biosynthesis from 200 mg/l to 874 mg/l. The increase was mainly due to activation of ruberitrinic acid biosynthesis. The expression of key AQ biosynthesis genes was strongly activated in rolA-transgenic calli. We compared the effects of the rolA gene with those of the rolB gene, which was previously considered the most potent inducer of secondary metabolism, and showed that rolA was more productive under conditions of long-term cultivation.

Increase of anthraquinone content in Rubia cordifolia cells transformed by native and constitutively active forms of the AtCPK1 gene.

KEY MESSAGE: Overexpression of both native and mutant forms of AtCPK1 in Rubia cordifolia cells increased anthraquinone production and transcript abundance of the RcIPPI, RcOSBL, RcOSBS , and RcICS genes to different extents. Calcium-dependent protein kinases (CDPKs) are involved in various cell processes and are regulated by a calcium signal system. CDPKs also function in plant defense against stress factors such as pathogens, temperature, and salinity. In this study, we compared the effect of heterologous expression of two forms of the Arabidopsis AtCPK1 gene, native and constitutively active (Ca(2+)-independent), on anthraquinone production in transgenic Rubia cordifolia cells. Significant qualitative and quantitative differences were found in the content of anthraquinone derivatives in control and AtCPK1-transgenic calli. Expression of the AtCPK1 gene increased anthraquinone production by 3 and 12 times for native and constitutively active forms, respectively, compared with control cells. In addition, we identified and quantified the expression of genes encoding key enzymes of the anthraquinone biosynthesis pathway, including isochorismate synthase (ICS), o-succinylbenzoate synthase (OSBS), o-succinylbenzoate ligase (OSBL), and isopentenyl diphosphate isomerase (IPPi). In all AtCPK1-transgenic cell lines, expression of ICS, OSBS, OSBL, and IPPi increased considerably at 14-15 days of subculture and decreased at the end of cultivation (30 days). The results suggest that both native and constitutively active AtCPK1 forms induced anthraquinone accumulation at the logarithmic growth stage via enhancement of expression of genes involved in the metabolism of anthraquinones or their regulatory mechanisms.

Genome-editing tools for stem cell biology.

Human pluripotent stem cells provide a versatile platform for regenerative studies, drug testing and disease modeling. That the expression of only four transcription factors, Oct4, Klf4, Sox2 and c-Myc (OKSM), is sufficient for generation of induced pluripotent stem cells (iPSCs) from differentiated somatic cells has revolutionized the field and also highlighted the importance of OKSM as targets for genome editing. A number of novel genome-editing systems have been developed recently. In this review, we focus on successful applications of several such systems for generation of iPSCs. In particular, we discuss genome-editing systems based on zinc-finger fusion proteins (ZFs), transcription activator-like effectors (TALEs) and an RNA-guided DNA-specific nuclease, Cas9, derived from the bacterial defense system against viruses that utilizes clustered regularly interspaced short palindromic repeats (CRISPR).