Evidence for the Presence of 5S rRNA in Mammalian Mitochondria

Mammalian mitochondrial ribosomes contain two prokaryotic-like rRNAs, 12S and 16S, both encoded by mitochondrial DNA. As opposed to cytosolic ribosomes, however, these ribosomes are not thought to contain 5S rRNA. For this reason, it has been unclear whether 5S rRNA, which can be detected in mitochondrial preparations, is an authentic organellar species imported from the cytosol or is merely a copurifying cytosol-derived contaminant. We now show that 5S rRNA is tightly associated with highly purified mitochondrial fractions of human and rat cells and that 5S rRNA transcripts derived from a synthetic gene transfected transiently into human cells are both expressed in vivo and present in highly purified mitochondria and mitoplasts. We conclude that 5S rRNA is imported into mammalian mitochondria, but its function there still remains to be clarified.

The only essential function of TFIIIA in yeast is the transcription of 5S rRNA genes.

We have developed a system to transcribe the yeast 5S rRNA gene in the absence of the transcription factor TFIIIA. A long transcript was synthesized both in vitro and in vivo from a hybrid gene in which the tRNA-like promoter sequence of the RPR1 gene was fused to the yeast 5S RNA gene. No internal initiation directed by the endogenous 5S rDNA promoter or any processing of the hybrid transcript was observed in vitro. Yeast cells devoid of transcription factor TFIIIA, which, therefore, could not synthesize any 5S rRNA from the endogenous chromosomal copies of 5S rDNA, could survive if they carried the hybrid RPR1-5S construct on a multicopy plasmid. In this case, the only source of 5S rRNA was the precursor RPR1-5S transcript that gave rise to two RNA species slightly larger than wild-type 5S rRNA. This establishes that the only essential function of TFIIIA is to promote the synthesis of 5S rRNA. However, cells devoid of TFIIIA and surviving with these two RNAs grew more slowly at 30 degrees C compared with wild-type cells and were thermosensitive at 37 degrees C.

The tRNA processing enzyme RNase T is essential for maturation of 5S RNA.

The maturation of 5S RNA in Escherichia coli is poorly understood. Although it is known that large precursors of 5S RNA accumulate in mutant cells lacking the endoribonuclease-RNase E, almost nothing is known about how the mature 5' and 3' termini of these molecules are generated. We have examined 5S RNA maturation in wild-type and single- or multiple-exoribonuclease-deficient cells by Northern blot and primer-extension analysis. Our results indicate that no mature 5S RNA is made in RNase T-deficient strains. Rather, 5S RNA precursors containing predominantly 2 extra nucleotides at the 3' end accumulate. Apparently, these 5S RNAs are functional inasmuch as mutant cells are viable, growing only slightly slower than wild type. Purified RNase T can remove the extra 3' residues, showing that it is directly involved in the trimming reaction. In contrast, mutations affecting other 3' exoribonucleases have no effect on 5S RNA maturation. Approximately 90% of the 5S RNAs in both wild-type and RNase T- cells contain mature 5' termini, indicating that 5' processing is independent of RNase T action. These data identify the enzyme responsible for generating the mature 3' terminus of 5S RNA molecules and also demonstrate that a completely processed 5S RNA molecule is not essential for cell survival.

Implementação da metodologia 5S no âmbito da utilização das ferramentas de corte

O trabalho apresentado surgiu com a necessidade da realização de um projeto de mestrado, em conjunto com a oportunidade de implementar novos métodos organizacionais na empresa Farmi-Reparações, Lda. Para efetuar esta alteração recorreu-se à metodologia 5S, tendo sido necessário para tal realizar um estudo teórico às ferramentas de melhoria da qualidade, com maior destaque para a metodologia 5S e o fator de resistência à mudança inerente à implementação. No sentido de alcançar os objetivos propostos, realizou-se uma análise inicial à empresa, e delineou-se uma estratégia para contornar a resistência à mudança por parte dos colaboradores aos métodos de trabalho, que poderiam colocar em causa a implementação da metodologia 5S. Com base na estratégia definida, aplicou-se a metodologia em alguns postos de trabalho da secção de produção, e na totalidade da secção de bancada. Para tal, foi necessário a empresa realizar alguns investimentos, nomeadamente, com a aquisição e fabricação de sistemas de organização. Ao nível da medição do impacto da implementação dos 5S, foi analisado o grau de aplicação de cada senso e identificados os benefícios obtidos na seção de bancada. Foi realizada ainda uma auditoria 5S e comparados os resultados das quebras de ferramentas de corte e saída por desgaste do 1º semestre de 2016 com o 1º semestre de 2015. Com base nos resultados e trabalho realizado, é de destacar que o impacto mais positivo deste trabalho foi o facto de este ter sido o início do processo de melhoria contínua na empresa, gerando as condições necessárias para implementar outras técnicas da qualidade, pois permitiu contornar a resistência à mudança dos colaboradores no que respeita aos métodos de trabalho e organizacionais estabelecidos.

Faster group operations on elliptic curves

This paper improves implementation techniques of Elliptic Curve Cryptography. We introduce new formulae and algorithms for the group law on Jacobi quartic, Jacobi intersection, Edwards, and Hessian curves. The proposed formulae and algorithms can save time in suitable point representations. To support our claims, a cost comparison is made with classic scalar multiplication algorithms using previous and current operation counts. Most notably, the best speeds are obtained from Jacobi quartic curves which provide the fastest timings for most scalar multiplication strategies benefiting from the proposed 12M + 5S + 1D point doubling and 7M + 3S + 1D point addition algorithms. Furthermore, the new addition algorithm provides an efficient way to protect against side channel attacks which are based on simple power analysis (SPA). Keywords: Efficient elliptic curve arithmetic,unified addition, side channel attack.

The AusD Study: A population-based study of the determinants of serum 25-hydroxyvitamin D concentration across a broad latitude range

Observational studies suggest that people with a high serum 25-hydroxyvitamin D (25(OH)D) concentration may have reduced risk of chronic diseases such as osteoporosis, multiple sclerosis, type 1 diabetes, cardiovascular disease, and some cancers. The AusD Study (A Quantitative Assessment of Solar UV Exposure for Vitamin D Synthesis in Australian Adults) was conducted to clarify the relationships between ultraviolet (UV) radiation exposure, dietary intake of vitamin D, and serum 25(OH)D concentration among Australian adults residing in Townsville (19.3°S), Brisbane (27.5°S), Canberra (35.3°S), and Hobart (42.8°S). Participants aged 18-75 years were recruited from the Australian Electoral Roll between 2009 and 2010. Measurements were made of height, weight, waist:hip ratio, skin, hair, and eye color, blood pressure, and grip strength. Participants completed a questionnaire on sun exposure and vitamin D intake, together with 10 days of personal UV dosimetry and an associated sun-exposure and physical-activity diary that was temporally linked to a blood test for measurement of 25(OH)D concentration. Ambient solar UV radiation was also monitored at all study sites. We collected comprehensive, high-quality data from 1,002 participants (459 males, 543 females) assessed simultaneously across a range of latitudes and through all seasons. Here we describe the scientific and methodological issues considered in designing the AusD Study.

A systematic approach for selecting lean strategies and assessing leanness in manufacturing organizations

Lean strategies have been developed to eliminate or reduce manufacturing waste and thus improve operational efficiency in manufacturing processes. However, implementing lean strategies requires a large amount of resources and, in practice, manufacturers encounter difficulties in selecting appropriate lean strategies within their resource constraints. There is currently no systematic methodology available for selecting appropriate lean strategies within a manufacturer's resource constraints. In the lean transformation process, it is also critical to measure the current and desired leanness levels in order to clearly evaluate lean implementation efforts. Despite the fact that many lean strategies are utilized to reduce or eliminate manufacturing waste, little effort has been directed towards properly assessing the leanness of manufacturing organizations. In practice, a single or specific group of metrics (either qualitative or quantitative) will only partially measure the overall leanness. Existing leanness assessment methodologies do not offer a comprehensive evaluation method, integrating both quantitative and qualitative lean measures into a single quantitative value for measuring the overall leanness of an organization. This research aims to develop mathematical models and a systematic methodology for selecting appropriate lean strategies and evaluating the leanness levels in manufacturing organizations. Mathematical models were formulated and a methodology was developed for selecting appropriate lean strategies within manufacturers' limited amount of available resources to reduce their identified wastes. A leanness assessment model was developed by using the fuzzy concept to assess the leanness level and to recommend an optimum leanness value for a manufacturing organization. In the proposed leanness assessment model, both quantitative and qualitative input factors have been taken into account. Based on program developed in MATLAB and C#, a decision support tool (DST) was developed for decision makers to select lean strategies and evaluate the leanness value based on the proposed models and methodology hence sustain the lean implementation efforts. A case study was conducted to demonstrate the effectiveness of these proposed models and methodology. Case study results suggested that out of 10 wastes identified, the case organization (ABC Limited) is able to improve a maximum of six wastes from the selected workstation within their resource limitations. The selected wastes are: unnecessary motion, setup time, unnecessary transportation, inappropriate processing, work in process and raw material inventory and suggested lean strategies are: 5S, Just-In-Time, Kanban System, the Visual Management System (VMS), Cellular Manufacturing, Standard Work Process using method-time measurement (MTM), and Single Minute Exchange of Die (SMED). From the suggested lean strategies, the impact of 5S was demonstrated by measuring the leanness level of two different situations in ABC. After that, MTM was suggested as a standard work process for further improvement of the current leanness value. The initial status of the organization showed a leanness value of 0.12. By applying 5S, the leanness level significantly improved to reach 0.19 and the simulation of MTM as a standard work method shows the leanness value could be improved to 0.31. The optimum leanness value of ABC was calculated to be 0.64. These leanness values provided a quantitative indication of the impacts of improvement initiatives in terms of the overall leanness level to the case organization. Sensitivity analsysis and a t-test were also performed to validate the model proposed. This research advances the current knowledge base by developing mathematical models and methodologies to overcome lean strategy selection and leanness assessment problems. By selecting appropriate lean strategies, a manufacturer can better prioritize implementation efforts and resources to maximize the benefits of implementing lean strategies in their organization. The leanness index is used to evaluate an organization's current (before lean implementation) leanness state against the state after lean implementation and to establish benchmarking (the optimum leanness state). Hence, this research provides a continuous improvement tool for a lean manufacturing organization.

Serum vitamin D levels are lower in Australian children and adolescents with type 1 diabetes than in children without diabetes

Vitamin D is synthesised in the skin through the action of UVB radiation (sunlight), and 25-hydroxy vitamin D (25OHD) measured in serum as a marker of vitamin D status. Several studies, mostly conducted in high latitudes, have shown an association between type 1 diabetes mellitus (T1DM) and low serum 25OHD. We conducted a case-control study to determine whether, in a sub-tropical environment with abundant sunlight (latitude 27.5°S), children with T1DM have lower serum vitamin D than children without diabetes. Fifty-six children with T1DM (14 newly diagnosed) and 46 unrelated control children participated in the study. Serum 25OHD, 1,25-dihydroxy vitamin D (1,25(OH)2D) and selected biochemical indices were measured. Vitamin D receptor (VDR) polymorphisms Taq1, Fok1, and Apa1 were genotyped. Fitzpatrick skin classification, self-reported daily hours of outdoor exposure, and mean UV index over the 35d prior to blood collection were recorded. Serum 25OHD was lower in children with T1DM (n=56) than in controls (n=46) [mean (95%CI)=78.7 (71.8-85.6) nmol/L vs. 91.4 (83.5-98.7) nmol/L, p=0.02]. T1DM children had lower self-reported outdoor exposure and mean UV exposure, but no significant difference in distribution of VDR polymorphisms. 25OHD remained lower in children with T1DM after covariate adjustment. Children newly diagnosed with T1DM had lower 1,25(OH)2D [median (IQR)=89 (68-122) pmol/L] than controls [121 (108-159) pmol/L, p=0.03], or children with established diabetes [137 (113-153) pmol/L, p=0.01]. Children with T1DM have lower 25OHD than controls, even in an environment of abundant sunlight. Whether low vitamin D is a risk factor or consequence of T1DM is unknown. © 2012 John Wiley & Sons A/S.

An E-Convergence Method For The Evaluation Of Critical Parameters

A simple cconversence technique is applied to obtain accurate estimates of critical temperatures and critical it\ponmts of a few two- and threpdiniensional king models. When applied to the virial series for hard spheres and hard discs, this method predicts a divergence of the equation-of-state at the density of closest packing.

On the number of two-line and three-line latin rectangles - an alternative approach

The number of two-line and three-line Latin rectangles is obtained by recursive methods in a setting slightly more general than usually considered. We show how this leads to a generalisation which is proved elsewhere.

Characterization of north-eastern Australian environments using APSIM for increasing rainfed maize production

Recurring water stresses are a major risk factor for rainfed maize cropping across the highly diverse agro-ecological environments of Queensland (Qld) and northern New South Wales (NNSW). Enhanced understanding of such agro-ecological diversity is necessary to more consistently sample target production environments for testing and targeting release of improved germplasm, and to improve the efficiency of the maize pre-breeding and breeding programs of Qld and New South Wales. Here, we used the Agricultural Production Systems Simulator (APSIM) – a well validated maize crop model to characterize the key distinctive water stress patterns and risk to production across the main maize growing regions of Qld and NNSW located between 15.8° and 31.5°S, and 144.5° and 151.8°E. APSIM was configured to simulate daily water supply demand ratios (SDRs) around anthesis as an indicator of the degree of water stress, and the final grain yield. Simulations were performed using daily climatic records during the period between 1890 and 2010 for 32 sites-soils in the target production regions. The runs were made assuming adequate nitrogen supply for mid-season maize hybrid Pioneer 3153. Hierarchical complete linkage analyses of the simulated yield resulted in five major clusters showing distinct probability distribution of the expected yields and geographic patterns. The drought stress patterns and their frequencies using SDRs were quantified using multivariate statistical methods. The identified stress patterns included no stress, mid-season (flowering) stress, and three terminal stresses differing in terms of severity. The combined frequency of flowering and terminal stresses was highest (82.9%), mainly in sites-soils combinations in the west of Qld and NNSW. Yield variability across the different sites-soils was significantly related to the variability in frequencies of water stresses. Frequencies of water stresses within each yield cluster tended to be similar, but different across clusters. Sites-soils falling within each yield cluster therefore could be treated as distinct maize production environments for testing and targeting newly developed maize cultivars and hybrids for adaptation to water stress patterns most common to those environments.

Purification and properties of a DNA polymerase from Mycobacterium tuberculosis H37Rv

DNA polymerase has been purified approximately 2000-fold from Mycobacterium tuberculosis H37Rv. The purified preparation was homogeneous by electrophoretic criteria and has a molecular weight of 135 000. The purified enzyme resembles Escherichia coli polymerase I in its properties, being insensitive to sulfhydryl drugs and possessing 5′,3′-exonuclease activity in addition to polymerase and 3′,5′-exonuclease activities. However, it differs from the latter in its sensitivity to higher salt concentration and DNA intercalating agents such as 8-aminoquinoline. The polymerase exhibited maximal activity between 37–42°C and pH 8.8–9.5. The polymerase was stable for several months below 0°C. However, the 5′,3′-exonuclease activity was more labile. The effects of different metal ions, polyamines and drugs on the polymerase activity are presented.

Catalyst effectiveness factor for redox model kinetic equation

A generalized isothermal effectiveness factor correlation has been proposed for catalytic reactions whose intrinsic kinetics are based on the redox model. In this correlation which is exact for asymptotic values of the Thiele parameter the effect of the parameters appearing in the model, the order of the reaction and particle geometry are incorporated in a modified form of Thiele parameter. The relationship takes the usual form: Image and predicts effectiveness factor with an error of less than 2% in a range of Thiele parameter that accommodates both the kinetic and diffusion control regimes.

Rate of ribonucleic acid chain growth in Mycobacterium tuberculosis H37Rv.

Two methods were employed to measure the rate of ribonucleic acid (RNA) chain growth in vivo in Mycobacterium tuberculosis H37Rv cultures growing in Sauton medium at 37 degrees C, with a generation time of 10 h. In the first, the bacteria were allowed to assimilate [3H]uracil or [3H]guanine into their RNA for short time periods. The RNA was then extracted and hydrolyzed with alkali, and the radioactivity in the resulting nucleotides and nucleosides was measured. The data obtained by this method allowed the calculation of the individual nucleotide step times during the growth of RNA chains, from which the average rate of RNA chain elongation was estimated to be about 4 nucleotides per s. The second method employed the antibiotic rifampin, which specifically inhibits the initiation of RNA synthesis without interfering with the elongation and completion of nascent RNA chains. Usint this method, the transcription time of the 16S, 23S, and 5S ribosomal RNA genes was estimated to be 7.6 min, which corresponds to a ribosomal RNA chain growth rate of 10 nucleotides per s.