Characterization of the monocyte-specific esterase (MSE) gene

Carboxylic esterases are widely distributed in hematopoietic cells. Monocytes express the esterase isoenzyme (termed 'monocyte-specific esterase', MSE) that can be inhibited by NaF in the alpha-naphthyl acetate cytochemical staining. We examined the expression of MSE in normal cells and primary and cultured leukemia-lymphoma cells. The MSE protein was demonstrated by isoelectric focusing (IEF); MSE mRNA expression was investigated by Northern blotting and reverse transcriptase-polymerase chain reaction (RT-PCR). The following samples were positive for MSE protein and Northern mRNA expression: 20/24 monocytic, 4/32 myeloid, and 1/20 erythroid-megakaryocytic leukemia cell lines, but none of the 112 lymphoid leukemia or lymphoma cell lines; of the normal purified cell populations only the monocytes were positive whereas, T, B cells, and granulocytes were negative; of primary acute (myelo) monocytic leukemia cells (CD14-positive, FAB M4/M5 morphology) 14/20 were Northern mRNA and 11/14 IEF protein positive. RT-PCR revealed MSE expression in 29/49 Northern-negative lymphoid leukemia-lymphoma cell lines. The RT-PCR signals in monocytic cell lines were on average 50-fold stronger than the mostly weak trace expression in lymphoid specimens. On treatment with various biomodulators, only all-trans retinoic acid significantly upregulated MSE message and protein levels but could not induce new MSE expression in several leukemia cell lines; lipopolysaccharide and interferon-gamma increased MSE expression in normal monocytes. Analysis of DNA methylation with sensitive restriction enzymes showed no apparent regulation of gene expression by differential methylation; the MSE gene is evolutionarily conserved among mammalian species; the half-life of the human MSE transcripts was about 5-6 h. The extent of MSE expression varied greatly among different monocytic leukemia samples. However, the MSE overexpression in a significant number of specimens was not associated with gene amplification, gross structural rearrangements or point mutations within the cDNA region. Taken together, the results suggest that MSE expression is not absolutely specific for, but strongly associated with cells of the monocytic lineage; MSE is either not expressed at all or expressed at much lower levels in cells from other lineages. The biological significance, if any, of rare MSE messages in lymphoid cells detectable only by the hypersensitive RT-PCR remains unclear. Further studies on the regulation of this gene and on the physiological function of the enzyme will no doubt be informative with respect to its striking overexpression in some malignant cells and to a possible role in the pathobiology of monocytic leukemias

Light-induced Fourier transform infrared (FTIR) spectroscopic investigations of the primary donor oxidation in bacterial photosynthesis

AbstractFourier transform infrared (FTIR) difference spectroscopy of the primary electron donor (P) photo-oxidation has been performed for reaction centers (RCs) and chromatophores of purple photosynthetic bacteria. In the 1800–650 cm−1 spectral region highly reproducible absorbance changes were obtained that can be related to specific changes of individual bond absorption. Several bands in the difference spectra are tentatively assigned to changes of intensity and position of the keto and ester CO vibrations of the P bacteriochlorophylls, and a possible interpretation in terms of changes of their environment or type of bonding to the protein is given. Small difference bands in the amide I and II region allow only minor protein conformational changes

Impact of Routine Platelet Reactivity Testing with VerifyNow Assay on Antiplatelet Choice After Percutaneous Coronary Intervention

Background: High on-treatment ADP platelet reactivity (HPR) measured by VerifyNow P2Y12 assay (VN) is an established risk factor for ischemic events after percutaneous coronary intervention (PCI). We hypothesized that routine use of VN at time of PCI in clinical practice may affect choice of P2Y12 antiplatelet therapy at discharge. Methods: In a single center retrospective analysis, we examined the influence of VN testing on choice of P2Y12 inhibitor post PCI in routine clinical practice. Assessment of HPR was used routinely in clinical care during the time period of analysis at discretion of clinical providers. Subjects with PRU>208 after the loading dose of clopidogrel or during clopidogrel steady state were switched to alternate P2Y12 inhibitors. Results: We identified 1001 patients with PCI during the time period specified. A total of 252 subjects underwent VN testing. Among those, 43% were found to have HPR on clopidogrel and were switched to alternate therapies (prasugrel [n=60], ticagrelor [n=48]). Patients who had VN platelet function testing were more likely to be discharged on clopidogrel as compared to those who did not have VN assay done (57% vs. 50%, p=0.039). There was no significant difference in 1-year net-MACE (CVD, MI, stent thrombosis, BARC 2 or higher bleeding) using tailored antiplatelet therapy (VN testing) as compared to standard of care group (adjusted HR:0.92, 95% CI: 0.54-1.5, p=0.74). Conclusion: Routine use of VN assay in personalized antiplatelet treatment decision-making after PCI is associated with lower likelihood of using novel P2Y12 inhibitors

Механические испытания элементов космической техники на синусоидальные воздействия

Целью работы является определение собственных частот трехмерной модели двигателя-маховика системы ориентации малого космического аппарата, а также ее поведение под действием квазистатической нагрузки, синусоидальных и случайных вибраций, акустического и ударного воздействия. Компьютерный анализ, проведенный в процессе проектирования, позволяет определить характеристики изделия и провести оптимизацию конструкции без создания опытного образца. Анализ проводился с помощью специального программного обеспечения "T-Flex Анализ". Исходная трехмерная модель была разбита на три подсборки: корпусную, роторную и статорную. Приведены результаты частотного анализа и расчета при воздействии на двигатель-маховик квазистатических нагрузок. По результатам проведенного компьютерного конечно-элементного анализа можно сказать, что спроектированный двигатель-маховик с большим запасом удовлетворяет требованиям надежности. Большие коэффициенты запасов говорят о том, что конструкцию в дальнейшем можно оптимизировать

Complete nucleotide sequence and structural organization of pPB1, a small Lactobacillus plantarum cryptic plasmid that originated by modular exchange

A small cryptic plasmid designated pPB1 was isolated from Lactobacillus plantarum BIFI-38 and its complete 2899 bp nucleotide sequence was determined. Sequence analysis revealed four putative open reading frames. Based on sequence analysis two modules could be identified. First, the replication module consisted of a sequence coding for a replication protein (RepB) and its corresponding target site, and two putative repressor proteins (RepA and RepC). Sequence analysis indicated the possible synthesis of an antisense RNA that might regulate RepB production. A putative laggingstrand initiation site was also found, suggesting that pPB1 replicates via a rolling circle mechanism. The second module of pPB1 consisted of a sequence coding for a putative mobilization protein and its corresponding oriT site. Since the nucleotide sequence of the replication module showed 94.5% identity to the similar region on the Leuconostoc lactis plasmid pCI411, and the nucleotide sequence of the mobilization module had 97.5% identity to L. plantarum plasmid pLB4, it is concluded that pPB1 originated by modular exchange between two such plasmids by homologous recombination. Putative recombination sites where crossover might have taken place were also identifiedThis work was supported by Grant 07G/0035/2003 from the Comunidad de Madrid and RM03-002 from the Instituto Nacional de Investigación y Tecnologı´a Agraria y Alimentaria (INIA). We thank R. Gonza´lez and A.V. Carrascosa for their advice and critical reading of the manuscript. B. de las Rivas was a recipient of a postdoctoral fellowship and A. Marcobal of a predoctoral fellowship both from the Comunidad de Madrid.Peer reviewe

Comet C/2011 W3 (Lovejoy): Orbit Determination, Outbursts, Disintegration of Nucleus, Dust-Tail Morphology, and Relationship to New Cluster of Bright Sungrazers

We describe the physical and orbital properties of C/2011 W3. After surviving perihelion, the comet underwent major changes (permanent loss of nuclear condensation, formation of spine tail). The process of disintegration culminated with an outburst on December 17.6 (T+1.6 d) and this delayed response is inconsistent with the rubble pile model. Probable cause was thermal stress from the heat pulse into the nucleus after perihelion, which could also produce fragmentation of sungrazers far from the Sun. The spine tail was a synchronic feature, made up of dust released at <30 m/s. Since the nucleus would have been located on the synchrone, we computed the astrometric positions of the missing nucleus as the coordinates of the points of intersection of the spine tail's axis with lines of forced orbital-period variation, derived from orbital solutions based on preperihelion astrometry from the ground. The resulting osculating orbital period was 698+/-2 years, which proves that C/2011 W3 is the first major member of the predicted new, 21st-century cluster of bright Kreutz-system sungrazers. The spine tail's tip contained dust 1-2 mm in diameter. The bizarre appearance of the dust tail in images taken hours after perihelion with coronagraphs on SOHO and STEREO is readily understood. The disconnection of the comet's head from the preperihelion tail and the apparent activity attenuation near perihelion are both caused by sublimation of all dust at heliocentric distances smaller than ~1.8 solar radii. The tail's brightness is strongly affected by forward scattering of sunlight by dust. The longest-imaged grains had a radiation-pressure parameter beta ~ 0.6, probably submicron-sized silicate grains. The place of C/2011 W3 within the hierarchy of the Kreutz system and its genealogy via a 14th century parent suggest that it is indirectly related to the celebrated sungrazer X/1106 C1.Comment: Submitted to Astrophysical Journal; 35 pages, 18 figures, 8 table

Tyrosyl-DNA phosphodiesterase 1 (TDP1) and its natural mutant scan1 inhibitors development as prototypes of drugs

In the present study, we performed screening of 21 compounds - derivatives of coumarin as inhibitors of tyrosyl-DNA phosphodiesterase 1 (Tdp1) and its mutant SCAN1. The ability of these compounds to inhibit Tdp1, important target for anti-cancer therapy, was studied. The most active derivatives have IC50 value of 0,1-1 mkM. SCAN1 inhibitors were found first in the world. They can likely underlie the development of drugs preventing or slowing the progressive cerebellar atrophy to improve quality of life of SCAN patients

Seasonal variations of glaciochemical, isotopic and stratigraphic properties in Siple Dome (Antarctica) surface snow

Six snow-pit records recovered from Siple Dome, West Antarctica, during 1994 are used to study seasonal variations in chemical (major ion and H202), isotopic (deuterium) and physical stratigraphic properties during the 1988-94 period. Comparison of δD measurements and satellite-derived brightness temperature for the Siple Dome area suggests that most seasonal SD maxima occur within ±4 weeks of each 1 January. Several other chemical species (H2O2, non-sea-salt (nss) SO4 2-, methanesulfonic acid and NO3-) show coeval peaks with SD, together providing an accurate method for identifying summer accumulation. Sea-salt-derived species generally peak during winter/spring, but episodic input is noted throughout some years. No reliable seasonal signal is identified in species with continental sources (nssCa2+ nss Mg2+), NH4 + or nssCl-. Visible strata such as large depth-hoar layers (\u3e5 cm) are associated with summer accumulation and its metamorphosis, but smaller hoar layers and crusts are more difficult to interpret. A multi-parameter approach is found to provide the most accurate dating of these snow-pit records, and is used to determine annual layer thicknesses at each site Significant spatial accumulation variability exists on an annual basis, but mean accumulation in the sampled 10 km2 grid for the 1988-94 period is fairly uniform