Interaction of acridine-calix[4]arene with DNA at the electrified liquid|liquid interface

The behaviour of an acridine-functionalised calix[4]arene at the interface between two immiscible electrolyte solutions (ITIES) is reported. Molecular modelling showed that the acridine-calix[4]arene has regions of significant net positive charge spread throughout the protonated acridine moieties, consistent with it being able to function as an anion ionophore. The presence of this compound in the organic phase facilitated the transfer of aqueous phase electrolyte ions. Upon addition of double stranded DNA to the aqueous phase, the transfer of electrolyte anions was diminished, due to DNA binding to the acridine moiety at the ITIES. The behaviour provides a basis for DNA hybridization detection using electrochemistry at the ITIES

Evolution of alternative and constitutive regions of mammalian 5'UTRs

<p>Abstract</p> <p>Background</p> <p>Alternative splicing (AS) in protein-coding sequences has emerged as an important mechanism of regulation and diversification of animal gene function. By contrast, the extent and roles of alternative events including AS and alternative transcription initiation (ATI) within the 5'-untranslated regions (5'UTRs) of mammalian genes are not well characterized.</p> <p>Results</p> <p>We evaluated the abundance, conservation and evolution of putative regulatory control elements, namely, upstream start codons (uAUGs) and open reading frames (uORFs), in the 5'UTRs of human and mouse genes impacted by alternative events. For genes with alternative 5'UTRs, the fraction of alternative sequences (those present in a subset of the transcripts) is much greater than that in the corresponding coding sequence, conceivably, because 5'UTRs are not bound by constraints on protein structure that limit AS in coding regions. Alternative regions of mammalian 5'UTRs evolve faster and are subject to a weaker purifying selection than constitutive portions. This relatively weak selection results in over-abundance of uAUGs and uORFs in the alternative regions of 5'UTRs compared to constitutive regions. Nevertheless, even in alternative regions, uORFs evolve under a stronger selection than the rest of the sequences, indicating that some of the uORFs are conserved regulatory elements; some of the non-conserved uORFs could be involved in species-specific regulation.</p> <p>Conclusion</p> <p>The findings on the evolution and selection in alternative and constitutive regions presented here are consistent with the hypothesis that alternative events, namely, AS and ATI, in 5'UTRs of mammalian genes are likely to contribute to the regulation of translation.</p

ВИВЧЕННЯ БУДОВИ ПРОДУКТУ ВЗАЄМОДІЇ ДИЛТІАЗЕМУ З БРОМКРЕЗОЛОВИМ ЗЕЛЕНИМ

The aim of the work.  Selection and identification of the diltiazem hydrochloride аnd bromocresol green reaction product.Materials and Methods. Diltiazem hydrochloride working standard, bromocresol green and the sample of final dosage forms were used.Reagents and solvents: A standard sample of diltiazem hydrochloride, bromocresol green, acetone.Analytical equipments: Spectrophotometer Specord Bruker Alpha (Bruker Optik GmbH, Ettlingen, Germany) using a prefix (full internal reflection) ATR (direct injection of substance), Spectrophotometer Specord 200 (Germany) (190-1100 nm).Results and Discussion. It was experimentally established that diltiazem hydrochloride forms coloured ion-pair product with bromocresol green in acetone medium. IR-spectrum ion-pair complex corresponds to the sum of absorption of starting compounds with some differences, which confirm its formation. In ion-pair product in the region 3600–3400 sm-1 strips of phenolic –ОН appear with less intensity, which can be explained by the transformation of bromocresol green into the quinone structure. The product also has no absorption in the region 2370 cm-1 (R3N+ Cl- in diltiazem hydrochloride), which is explained by the formation of the ionic associate with bromocresol green, namely, the absorption is manifested in the form of a wider signal in the region 3200–2000 cm-1. Conclusions. Presented analytical assay showed that diltiazem hydrochloride reacts with bromocresol green in a ratio of 1:1. It was also established and identified the structure of the product of reaction. Using IR- spectroscopy, it was confirmed that is known to be an ionic associate.Мета роботи. Виділення та ідентифікація продукту взаємодії дилтіазему гідрохлориду з бромкрезоловим зеленим.Матеріали і методи. У дослідженні використано робочий стандартний зразок дилтіазему гідрохлориду, бромкрезоловий зелений (БКЗ), зразки готових лікарських форм вітчизняного та закордонного виробництва.Реагенти і розчинники: стандартний зразок дилтіазему гідрохлориду, бромкрезоловий зелений, ацетон.Аналітичне обладнання: спектрофотометр Bruker Alpha (Bruker Optik GmbH, Ettlingen, Germany) з використанням приставки (повного внутрішнього відбиття) ATR (пряме уведення речовини), спектрофотометр Specord 200 (Німеччина) (190–1100 нм).Результати й обговорення. Експериментально встановлено, що дилтіазем гідрохлорид взаємодіє з бромкрезоловим зеленим у середовищі ацетону з утворенням йонного асоціату. ІЧ-спектр йонного асоціату відповідає сумі поглинань вихідних сполук із деякими відмінностями, що підтверджують його утворення. У йонному асоціаті в області 3600–3400 см-1 смуги фенольних –ОН  проявляються з меншою інтенсивністю, що можна пояснити перетворенням БКЗ у хінонну структуру. У продукті також відсутні поглинання в області 2370 см-1  (R3N+ Cl- в дилтіаземі гідрохлориді), що пояснюється утворенням йонного асоціату з БКЗ, а саме поглинання проявляється у вигляді уширеного сигналу в області 3200–2000 см-1.Висновки. У результаті проведених досліджень встановлено, що дилтіазем гідрохлорид взаємодіє з бромкрезоловим зеленим у співвідношенні 1:1, виділено та встановлено будову продукту взаємодії дилтіазему гідрохлорид із бромкрезоловим зеленим. За допомогою ІЧ-спектроскопії підтверджено, що продуктом реакції є йонний асоціат

ІДЕНТИФІКАЦІЯ БУДОВИ ПРОДУКТУ ВЗАЄМОДІЇ ВЕРАПАМІЛУ ГІДРОХЛОРИДУ З БРОМКРЕЗОЛОВИМ ЗЕЛЕНИМ

The aim of the work. Isolation and identification of the product of interaction of verapamil hydrochloride with bromocresol green by IR spectrophotometry and 1H NMR spectroscopy methods. Materials and Methods. Verapamil hydrochloride working standard, bromocresol green and the sample of final dosage forms of Ukrainian and foreign production were used. Reagents and solvents: A standard sample of verapamil hydrochloride, bromocresol green, acetone. Analytical equipments: Spectrophotometer Specord Bruker Alpha (Bruker Optik GmbH, Ettlingen, Germany) using a prefix (full internal reflection) ATR (direct injection of substance), Spectrophotometer Specord 200(Germany) (190-1100 nm). Results and Discussion. It was experimentally established that verapamil hydrochloride forms coloured ion-pair product with bromocresol green in acetone medium. IR-spectrum ion-pair complex corresponds to the sum of absorption of starting compounds with some differences, which confirm its formation. In ion-pair product in the region 3600‒3400 cm‒1 strips of phenolic –ОН appear with less intensity, which can be explained by the transformation of bromocresol green into the quinone structure. The product also has no absorption in the region 2370cm‒1(R3N+Cl- in verapamil hydrochloride), which is explained by the formation of the ionic associate with bromocresol green, namely, the absorption is manifested in the form of a wider signal in the region 3200‒2000cm‒1. Conclusions. Presented analytical assay showed that verapamil hydrochloride reacts with bromocresol green in a ratio of 1:1, it was also established and identified the structure of the product of reaction. Using IR- spectroscopy and nuclear magnetic resonance spectrometry, it was confirmed that is known to be an ionic pair.Мета роботи. Виділення та ідентифікація продукту взаємодії верапамілу гідрохлориду з бромкрезоловим зеленим методами ІЧ-спектрофотометрії та 1H ЯMР спектроскопії. Матеріали і методи. У дослідженні використано робочий стандартний зразок верапамілу гідрохлориду, бромкрезоловий зелений (БКЗ), взірці готових лікарських форм вітчизняного та зарубіжного виробництва. Реагенти і розчинники: стандартний зразок верапамілу гідрохлориду, бромкрезоловий зелений, ацетон. Аналітичне обладнання: Спектрофотометр Bruker Alpha (Bruker Optik GmbH, Ettlingen, Germany) з використанням приставки (повного внутрішнього відбиття) ATR (пряме уведення речовини), спектрофотометр Specord 200 (Німеччина) (190–1100 нм). Результати й обговорення. Експериментально встановлено, що верапамілу гідрохлорид взаємодіє з бромкрезоловим зеленим у середовищі ацетону з утворенням йонного асоціату. ІЧ-спектр йонного асоціату відповідає сумі поглинань вихідних сполук із деякими відмінностями, що підтверджують його утворення. У йонному асоціаті в області 3600‒3400 см‒1 смуги фенольних –ОН проявляються з меншою інтенсивністю, що можна пояснити перетворенням БКЗ у хінонну структуру. У продукті також відсутні поглинання в області 2370см‒1 (R3N+Cl- у верапамілі гідрохлориді), що пояснюється утворенням йонного асоціату з БКЗ, а саме поглинання проявляється у вигляді уширеного сигналу в області 3200‒2000см‒1. Висновки. У результаті проведених досліджень встановлено, що верапамілу гідрохлорид взаємодіє з бромкрезоловим зеленим у співвідношенні 1:1, виділено та встановлено будову забарвленого продукту взаємодії верапамілу гідрохлориду з бромкрезоловим зеленим. За допомогою методів ІЧ- спектроскопії та спектрометрії ядерного магнітного резонансу підтверджено, що продуктом взаємодії є йонний асоціат

The Bending Magnets for the Proton Transfer Line of CNGS

The project "CERN neutrinos to Gran Sasso (CNGS)", a collaboration between CERN and the INFN (Gran Sasso Laboratory) in Italy, will study neutrino oscillations in a long base-line experiment. High-energy protons will be extracted from the CERN SPS accelerator, transported through a 727 m long transfer line and focused onto a graphite target to produce a beam of pions and kaons and subsequently neutrinos. The transfer line requires a total of 78 dipole magnets. They were produced in the framework of an in-kind contribution of Germany via DESY to the CNGS project. The normal conducting dipoles, built from laminated steel cores and copper coils, have a core length of 6.3 m, a 37 mm gap height and a nominal field range of 1.38 T - 1.91 T at a maximum current of 4950 A. The magnet design was a collaboration between CERN and BINP. The half-core production was subcontracted to EFREMOV Institute; the coil fabrication, magnet assembly and the field measurements were concluded at BINP in June 2004. The main design issues and results of the acceptance tests, including mechanical, electrical and magnetic field measurements, are discussed

Levels of chemokines and other inflammatory mediators in patients with mild cognitive impairment undergoing rehabilitation

Alzheimer's disease is the most common neurodegenerative disease in old age. In some cases, it is preceded by mild cognitive impairment (MCI). One of the important components in the pathogenesis of neurodegeneration is chronic neuroinflammation (inflammatory activation of microglia and astrocytes in the brain). Systemic inflammatory response and immune dysregulation may contribute to neuroinflammation. The purpose of this study was to investigate the level of chemokines and other inflammatory mediators in patients with MCI who underwent medical rehabilitation, and to study its associations with the severity of cognitive impairment. The study group included 48 patients with MCI undergoing rehabilitation. Rehabilitation included cognitive therapy, psychotherapy and tasks for unaided performance. Repeated examination was conducted 6 months after the completion of rehabilitation. The control group included 46 healthy volunteers. Multiplex assay was used to determine serum cytokine and chemokine concentrations. Student's t-test was used to assess the significance of differences. Assessment of cognitive functions was performed using international neuropsychological scales. In patients with MCI, we have found an increase in the levels of several cytokines and chemokines (TNFα, CXCL10/IP10, MDC) that regulate systemic inflammation, cellular and humoral mechanisms of adaptive immunity. After the rehabilitation course their levels returned to normal. It was also found that decrease in CCL7 level in the patients before the rehabilitation course is associated with the severity of cognitive impairment. The findings contribute to understanding the role of chemokines in the pathogenesis of MCI, and indicate that their levels can be potential biomarkers of the severity of cognitive impairment. For translation of the findings into clinical practice, their validation in larger studies is needed, as well as assessing the associations between chemokine levels and the severity of cognitive impairment in MCI over long-term follow-up

Expression Patterns of Protein Kinases Correlate with Gene Architecture and Evolutionary Rates

Protein kinase (PK) genes comprise the third largest superfamily that occupy ∼2% of the human genome. They encode regulatory enzymes that control a vast variety of cellular processes through phosphorylation of their protein substrates. Expression of PK genes is subject to complex transcriptional regulation which is not fully understood.Our comparative analysis demonstrates that genomic organization of regulatory PK genes differs from organization of other protein coding genes. PK genes occupy larger genomic loci, have longer introns, spacer regions, and encode larger proteins. The primary transcript length of PK genes, similar to other protein coding genes, inversely correlates with gene expression level and expression breadth, which is likely due to the necessity to reduce metabolic costs of transcription for abundant messages. On average, PK genes evolve slower than other protein coding genes. Breadth of PK expression negatively correlates with rate of non-synonymous substitutions in protein coding regions. This rate is lower for high expression and ubiquitous PKs, relative to low expression PKs, and correlates with divergence in untranslated regions. Conversely, rate of silent mutations is uniform in different PK groups, indicating that differing rates of non-synonymous substitutions reflect variations in selective pressure. Brain and testis employ a considerable number of tissue-specific PKs, indicating high complexity of phosphorylation-dependent regulatory network in these organs. There are considerable differences in genomic organization between PKs up-regulated in the testis and brain. PK genes up-regulated in the highly proliferative testicular tissue are fast evolving and small, with short introns and transcribed regions. In contrast, genes up-regulated in the minimally proliferative nervous tissue carry long introns, extended transcribed regions, and evolve slowly.PK genomic architecture, the size of gene functional domains and evolutionary rates correlate with the pattern of gene expression. Structure and evolutionary divergence of tissue-specific PK genes is related to the proliferative activity of the tissue where these genes are predominantly expressed. Our data provide evidence that physiological requirements for transcription intensity, ubiquitous expression, and tissue-specific regulation shape gene structure and affect rates of evolution

Structural Conformers of (1,3-Dithiol-2-ylidene)ethanethioamides: The Balance Between Thioamide Rotation and Preservation of Classical Sulfur-Sulfur Hypervalent Bonds

The reaction of N-(2-phthalimidoethyl)-N-alkylisopropylamines and S2Cl2 gave 4-N-(2-phthalimidoethyl)-N-alkylamino-5-chloro-1,2-dithiol-3-thiones that quantitatively cycloadded to dimethyl or diethyl acetylenedicarboxylate to give stable thioacid chlorides, which in turn reacted with one equivalent of aniline or a thiole to give thioanilides or a dithioester. Several compounds of this series showed atropisomers that were studied by a combination of dynamic NMR, simulation of the signals, conformational analysis by DFT methods, and single crystal X-ray diffraction, showing a good correlation between the theoretical calculations, the experimental values of energies, and the preferred conformations in the solid state. The steric hindering of the crowded substitution at the central amine group was found to be the reason for the presence of permanent atropisomers in this series of compounds and the cause of a unique disposition of the thioxo group at close-to-right angles with respect to the plane defined by the 1,3-dithiole ring in the dithiafulvene derivatives, thus breaking the sulfur–sulfur hypervalent bond that is always found in this kind of compounds.Ministerio de Economıá y Competitividad, Spain (Project CTQ2012- 31611), Junta de Castilla y León, Consejería de Educación y Cultura y Fondo Social Europeo (Project BU246A12-1), and the European Commission, Seventh Framework Programme (Project SNIFFER FP7-SEC-2012-312411

Multispectral analysis of Northern Hemisphere temperature records over the last five millennia

Aiming to describe spatio-temporal climate variability on decadal-to-centennial time scales and longer, we analyzed a data set of 26 proxy records extending back 1,000–5,000 years; all records chosen were calibrated to yield temperatures. The seven irregularly sampled series in the data set were interpolated to a regular grid by optimized methods and then two advanced spectral methods—namely singular-spectrum analysis (SSA) and the continuous wavelet transform—were applied to individual series to separate significant oscillations from the high noise background. This univariate analysis identified several common periods across many of the 26 proxy records: a millennial trend, as well as oscillations of about 100 and 200 years, and a broad peak in the 40–70-year band. To study common NH oscillations, we then applied Multichannel SSA. Temperature variations on time scales longer than 600 years appear in our analysis as a dominant trend component, which shows climate features consistent with the Medieval Warm Period and the Little Ice Age. Statistically significant NH-wide peaks appear at 330, 250 and 110&nbsp;years, as well as in a broad 50–80-year band. Strong variability centers in several bands are located around the North Atlantic basin and are in phase opposition between Greenland and Western Europe