Fine Tuning of the Lactate and Diacetyl Production through Promoter Engineering in Lactococcus lactis

Lactococcus lactis is a well-studied bacterium widely used in dairy fermentation and capable of producing metabolites with organoleptic and nutritional characteristics. For fine tuning of the distribution of glycolytic flux at the pyruvate branch from lactate to diacetyl and balancing the production of the two metabolites under aerobic conditions, a constitutive promoter library was constructed by randomizing the promoter sequence of the H2O-forming NADH oxidase gene in L. lactis. The library consisted of 30 promoters covering a wide range of activities from 7,000 to 380,000 relative fluorescence units using a green fluorescent protein as reporter. Eleven typical promoters of the library were selected for the constitutive expression of the H2O-forming NADH oxidase gene in L. lactis, and the NADH oxidase activity increased from 9.43 to 58.17-fold of the wild-type strain in small steps of activity change under aerobic conditions. Meanwhile, the lactate yield decreased from 21.15±0.08 mM to 9.94±0.07 mM, and the corresponding diacetyl production increased from 1.07±0.03 mM to 4.16±0.06 mM with the intracellular NADH/NAD+ ratios varying from 0.711±0.005 to 0.383±0.003. The results indicated that the reduced pyruvate to lactate flux was rerouted to the diacetyl with an almost linear flux variation via altered NADH/NAD+ ratios. Therefore, we provided a novel strategy to precisely control the pyruvate distribution for fine tuning of the lactate and diacetyl production through promoter engineering in L. lactis. Interestingly, the increased H2O-forming NADH oxidase activity led to 76.95% lower H2O2 concentration in the recombinant strain than that of the wild-type strain after 24 h of aerated cultivation. The viable cells were significantly elevated by four orders of magnitude within 28 days of storage at 4°C, suggesting that the increased enzyme activity could eliminate H2O2 accumulation and prolong cell survival

Деловая карьера и ее развитие в Территориальном органе Федеральной службы государственной статистики по Томской области(Росстат)

Выпускная квалификационная работа: 103 с., 7 рис., 3 табл. 51 источника, 7прил. Ключевые слова: делова карьера, персонал, организация, управление персоналом Объектом исследования является - организация управления персоналом в государственном учреждении. Цель работы - изучение и анализ механизмов управления карьерой и карьерными процессами на уровне индивида и организации с помощью теоретических знаний о сущности, видах, субъектах и методах управления деловой карьерой, а также выработке практических рекомендаций по индивидуальному управлению карьерой и карьерными процессами в исследуемой организации. В процессе исследования проводилось социологическое исследование. В результате исследования были разработаны несколько рекомендаций по совершенствованию деловой карьеры персонала ТомсFinal qualifying work: 103, Figure 7, Table 3... 51 source 7pril. Keywords: business career, personnel, organization, human resource management The object of this study is (are) - the organization of human resources management in the public institution. The purpose of work - study and analysis of the mechanisms of career and career management processes at the level of the individual and the organization wit h the help of theoretical knowledge about the nature, forms, subjects and business career management practices and practical recommendations for individual career management and career processes in the studied organizations. The study was conducted sociological research. The study several recommendations for improving the business caree

Autoantikörpernachweis mittels indirekter Immunfluoreszenz an HEp-2-Zellen

Aktuelle Diagnostik & Therapie | Review article Labormedizin Dtsch med Wochenschr 2009; 134(24): 1278-1282 DOI: 10.1055/s-0029-1225278 © Georg Thieme Verlag KG Stuttgart · New York Autoantikörpernachweis mittels indirekter Immunfluoreszenz an HEp-2-Zellen Autoantibody detection by indirect immunofluorescence on HEp-2 cells U. Sack1, K. Conrad2, E. Csernok3, I. Frank4, F. Hiepe5, T. Krieger6, A. Kromminga7, P von Landenberg8, G. Messer9, T. Witte10, R. Mierau11, für die deutsche EASI-Gruppe (European Autoimmunity Standardization Initiative) 1 Institut für Klinische Immunologie und Transfusionsmedizin, Medizinische Fakultät der Universität Leipzig 2 Institut für Immunologie, Medizinische Fakultät der Technischen Universität Dresden 3 Poliklinik für Rheumatologie, Universitätsklinikum Schleswig-Holstein, Campus Lübeck 4 Labor Dr. Tiller, München 5 Medizinische Klinik mit Schwerpunkt Rheumatologie und Klinische Immunologie, Charité, Berlin 6 Institut für Immunologie, Universitätsklinikum Eppendorf der Universität Hamburg 7 Labor Lademannbogen, Hamburg 8 Institut für Laboratoriumsmedizin, Universität Mainz 9 Klinik und Poliklinik für Dermatologie und Allergologie, Ludwig-Maximilians-Universität München 10 Institut für Klinische Immunologie und Rheumatologie, Medizinische Hochschule Hannover 11 Labor an der Rheumaklinik Aachen Zusammenfassung Der Nachweis von Autoantikörpern ist ein wichtiger Bestandteil der Diagnostik und Verlaufskontrolle von Patienten mit Autoimmunerkrankungen. In der Labordiagnostik der Kollagenosen und der autoimmunen Lebererkrankungen spielt die indirekte Immunfluoreszenz an HEp-2-Zellen im Rahmen einer Stufendiagnostik eine zentrale Rolle. Trotz hoher Qualität der Diagnostik können die Befunde zwischen verschiedenen Labors jedoch wegen fehlender Standardisierung und subjektiver Faktoren zum Teil erheblich differieren. Die vorliegende Arbeit formuliert Empfehlungen für eine vereinheitlichte Bearbeitung und Interpretation des HEp-2-Zell-Testes zum Nachweis von nicht organspezifischen (v. a. antinukleären) Antikörpern. Dabei werden Anforderungen an die verwendeten Diagnostika genannt, sowie Hinweise für die Abarbeitung und Auswertung im Labor und Empfehlungen für die Interpretation gegeben. Für eine optimale Labordiagnostik sind neben einer aussagefähigen klinischen Verdachtsdiagnose und einem erfahrenen Labordiagnostiker folgende Eckpunkte zu empfehlen: Initiales Screening mittels indirekter Immunfluoreszenz an sorgfältig ausgewählten HEp2-Zellen, beginnend mit einer Serumverdünnung von 1 : 80 und Auswertung in einem lichtstarken Mikroskop, positive Befundung ab einem Titer von 1 : 160, laborinterne Qualitätssicherung und vereinheitlichte Befundung. Ziel ist die Verbesserung der Diagnostik und Betreuung von Patienten mit Autoimmunerkrankungen als zentrales Anliegen der „European Autoimmunity Standardization Initiative” (EASI). Summary Systemic autoimmune diseases are characterized by the presence of antinuclear autoantibodies (ANA). Diluted patient sera are typically used to screen for the presence of ANA by immunfluorescence microscopy with fixed HEp-2 cells. Despite high-quality test kits, reports of different laboratories frequently present controversial results. This article recommends unified processing and interpretation of HEp-2 based screening for autoantibodies. Suggestions are made for the selection of high-quality test kits, optimized processing and diagnostic procedures. In addition to a relevant clinical diagnosis and an experienced laboratory specialist, the following procedure is highly recommended to achieve good laboratory practice: Initial HEp-2 based screening by indirect immunofluorescence, starting with a 1:80 serum dilution, and evaluation in a bright fluorescence microscope, pathological values from a titer of 1:160 upwards, internal quality checks and unified interpretation. We aim to improve diagnosis and care of patients with autoimmune diseases as a central focus of the European Autoimmunity Standardization Initiative (EASI)

The Impact of Age on the Association Between Physical Activity and White Matter Integrity in Cognitively Healthy Older Adults

Cognition emerges from coordinated processing among distributed cortical brain regions, enabled through interconnected white matter networks. Cortical disconnection caused by age-related decline in white matter integrity (WMI) is likely to contribute to age-related cognitive decline. Physical activity (PA) has been suggested to have beneficial effects on white matter structure. However, its potential to counteract age-related decline in WMI is not yet well established. The present explorative study analyzed if PA was associated with WMI in cognitively healthy older adults and if this association was modulated by age. Forty-four cognitively healthy older individuals (aged 60–88 years) with diffusion-tensor imaging (DTI) and PA measurements were included from the AgeGain study. Voxelwise analysis using Tract-Based Spatial Statistics (TBSS) demonstrated that PA was associated with WMI in older adults. However, results emphasized that this association was restricted to high age. The association between PA and WMI was found in widespread white matter regions suggesting a global rather than a regional effect. Supplementary analyses demonstrated an association between the integrity of these regions and the performance in memory [verbal learning and memory test (VLMT)] and executive functioning (Tower of London).Results of the present explorative study support the assumption that PA is associated with WMI in older adults. However, results emphasize that this association is restricted to high age. Since cognitive decline in the elderly is typically most pronounced in later stages of aging, PA qualifies as a promising tool to foster resilience against age-related cognitive decline, via the preservation of the integrity of the brains WM

Dorsolateral Prefrontal Functional Connectivity Predicts Working Memory Training Gains

BACKGROUND: Normal aging is associated with working memory decline. A decrease in working memory performance is associated with age-related changes in functional activation patterns in the dorsolateral prefrontal cortex (DLPFC). Cognitive training can improve cognitive performance in healthy older adults. We implemented a cognitive training study to assess determinants of generalization of training gains to untrained tasks, a key indicator for the effectiveness of cognitive training. We aimed to investigate the association of resting-state functional connectivity (FC) of DLPFC with working memory performance improvement and cognitive gains after the training. METHODS: A sample of 60 healthy older adults (mean age: 68 years) underwent a 4-week neuropsychological training, entailing a working memory task. Baseline resting-state functional MRI (rs-fMRI) images were acquired in order to investigate the FC of DLPFC. To evaluate training effects, participants underwent a neuropsychological assessment before and after the training. A second follow-up assessment was applied 12 weeks after the training. We used cognitive scores of digit span backward and visual block span backward tasks representing working memory function. The training group was divided into subjects who had and who did not have training gains, which was defined as a higher improvement in working memory tasks than the control group (N = 19). RESULTS: A high FC of DLPFC of the right hemisphere was significantly associated with training gains and performance improvement in the visuospatial task. The maintenance of cognitive gains was restricted to the time period directly after the training. The training group showed performance improvement in the digit span backward task. CONCLUSION: Functional activation patterns of the DLPFC were associated with the degree of working memory training gains and visuospatial performance improvement. Although improvement through cognitive training and acquisition of training gains are possible in aging, they remain limited