Functional characterization of the Pneumocystis jirovecii potential drug targets dhfs and abz2 involved in folate biosynthesis.

Pneumocystis species are fungal parasites colonizing mammal lungs with strict host specificity. Pneumocystis jirovecii is the human-specific species and can turn into an opportunistic pathogen causing severe pneumonia in immunocompromised individuals. This disease is currently the second most frequent life-threatening invasive fungal infection worldwide. The most efficient drug, cotrimoxazole, presents serious side effects, and resistance to this drug is emerging. The search for new targets for the development of new drugs is thus of utmost importance. The recent release of the P. jirovecii genome sequence opens a new era for this task. It can now be carried out on the actual targets to be inhibited instead of on those of the relatively distant model Pneumocystis carinii, the species infecting rats. We focused on the folic acid biosynthesis pathway because (i) it is widely used for efficient therapeutic intervention, and (ii) it involves several enzymes that are essential for the pathogen and have no human counterparts. In this study, we report the identification of two such potential targets within the genome of P. jirovecii, the dihydrofolate synthase (dhfs) and the aminodeoxychorismate lyase (abz2). The function of these enzymes was demonstrated by the rescue of the null allele of the orthologous gene of Saccharomyces cerevisiae

Genetic and environmental influences on in vitro digestibility of alfalfa

To study the relationships between in vitro NDF or true dry matter digestibility (NDFD and IVTDMD) and forage fibre content, 95 alfalfa samples from 5 cultivars grown in 3 different locations and from different cuts were analysed for fibre fractions and evaluated for NDFD and IVTDMD. NDFD was mainly influenced by order of cut and age of the fields. The multifoliate cultivar controlled did not appear to differ for fibre composition and NDFD from the other ones. Fibre fractions contents explained a large part of the difference in NDFD and IVTDMD, and no significant differences in this relationships appeared for first-cut vs. aftermath forage. The best predictive equation of NDFD from fibre factions was: NDFD = 73.61 -0.62 * NDF (% DM) – 56.33 * ADL/NDF (R2=0.39). Variations in fibre content and quality do not completely explain differences in NDFD suggesting the interference of other factors that are worth to be better studied

Oneiric stress and safety and security at work: the discovery of a new universal symbol

Cox and Griffiths define as psychosocial risks at work “those aspects of the planning, organization and management of work, which, along with their environmental and social contexts, may affect mental and physical health of the employees, directly or indirectly producing stress”. Therefore, a more effective approach to occupational safety and security should include integrated risk management through the identification of any work stress related problem. The purpose of this paper is to analyze the possible correlation of risk at work with the modification of sleep, and inside it, the specific function of dream activity

Unveiling the complexity of japanese metallic threads

In the framework of an extensive survey campaign on a collection of Japanese samurai armors, metallic threads from different parts of the traditional equipment were studied by several analytical techniques. The collection of armors belongs to Museo delle Culture (Lugano, Switzerland) and it is composed of ten elements, which date back from the 15th to 20th century. Metallic threads under study come from six of ten elements of the collection and represent a complex and unique multimaterial, which shows specific characteristics in Japanese tradition (kinran). The multianalytical approach based on ATR-FTIR spectroscopy and SEM-EDX analysis, together with a careful observation with optical and digital microscopy, permitted to obtain a complete characterization of materials, which have shown a great variability in metal foils and in organic adhesives (urushi, animal glue, starch). Gold and silver turned out to be not so largely used as scholars thought, while aluminum showed a great diffusion. Within the collection of analyzed armors, the obtained results allowed us for the first time to get a complete comprehension of materials and techniques used by Japanese craftsmen, and to observe differences in the quality of the materials and in manufacture technology over the centuries

Assessment of GNC Impacts of Chemical Plume Impingement in the Case of Prisma Irides Experiment

This paper presents the preliminary analysis of an in-orbit demonstration opportunity to test plume impingement as a viable means to change the attitude state of a space debris based on the Prisma and Picard missions. This technique has been proposed as part of the COBRA concept studied by ESA in collaboration with GMV, Politecnico di Milano and Thales-Alenia Space, as an active debris removal concept relying on the exhaust plume of a monopropellant chemical propulsion system as a means to impart momentum and ultimately modify the orbit of a space debris object in a contactless manner. The feasibility of the experiment is presented as well as its critical areas, no showstoppers are identified

Mechanisms of Surface Antigenic Variation in the Human Pathogenic Fungus <i>Pneumocystis jirovecii</i>.

Microbial pathogens commonly escape the human immune system by varying surface proteins. We investigated the mechanisms used for that purpose by &lt;i&gt;Pneumocystis jirovecii&lt;/i&gt; This uncultivable fungus is an obligate pulmonary pathogen that in immunocompromised individuals causes pneumonia, a major life-threatening infection. Long-read PacBio sequencing was used to assemble a core of subtelomeres of a single &lt;i&gt;P. jirovecii&lt;/i&gt; strain from a bronchoalveolar lavage fluid specimen from a single patient. A total of 113 genes encoding surface proteins were identified, including 28 pseudogenes. These genes formed a subtelomeric gene superfamily, which included five families encoding adhesive glycosylphosphatidylinositol (GPI)-anchored glycoproteins and one family encoding excreted glycoproteins. Numerical analyses suggested that diversification of the glycoproteins relies on mosaic genes created by ectopic recombination and occurs only within each family. DNA motifs suggested that all genes are expressed independently, except those of the family encoding the most abundant surface glycoproteins, which are subject to mutually exclusive expression. PCR analyses showed that exchange of the expressed gene of the latter family occurs frequently, possibly favored by the location of the genes proximal to the telomere because this allows concomitant telomere exchange. Our observations suggest that (i) the &lt;i&gt;P. jirovecii&lt;/i&gt; cell surface is made of a complex mixture of different surface proteins, with a majority of a single isoform of the most abundant glycoprotein, (ii) genetic mosaicism within each family ensures variation of the glycoproteins, and (iii) the strategy of the fungus consists of the continuous production of new subpopulations composed of cells that are antigenically different. &lt;b&gt;IMPORTANCE&lt;/b&gt; &lt;i&gt;Pneumocystis jirovecii&lt;/i&gt; is a fungus causing severe pneumonia in immunocompromised individuals. It is the second most frequent life-threatening invasive fungal infection. We have studied the mechanisms of antigenic variation used by this pathogen to escape the human immune system, a strategy commonly used by pathogenic microorganisms. Using a new DNA sequencing technology generating long reads, we could characterize the highly repetitive gene families encoding the proteins that are present on the cellular surface of this pest. These gene families are localized in the regions close to the ends of all chromosomes, the subtelomeres. Such chromosomal localization was found to favor genetic recombinations between members of each gene family and to allow diversification of these proteins continuously over time. This pathogen seems to use a strategy of antigenic variation consisting of the continuous production of new subpopulations composed of cells that are antigenically different. Such a strategy is unique among human pathogens

Differentiating between Natural and Modified Cellulosic Fibres Using ATR-FTIR Spectroscopy

This paper presents the limitations and potential of ATR-FTIR spectroscopy applied to the study of cellulosic textile collections The technique helps to differentiate natural fibres according to the content of lignin, pectin, hemicellulose, and wax, although some problematic issues should be considered. The spectral differences derived from the environmental humidity uptake and the plant composition are reviewed and discussed in the light of new experimental data. Diagnostic bands are proposed that can discriminate between different fibres from different plants. The contribution of ageing is also considered, demonstrating that sometimes aged fibres cannot be reliably recognised. In contrast, the potential of ATR-FTIR spectroscopy to discriminate between natural and modified fibres is discussed and proven. The best results were obtained when microinvasive ATR-FTIR spectroscopy was coupled with SEM observations. The proposed protocol was tested on microsamples of various cellulosic materials from traditional Japanese samurai armours dating from the 16th to the 20th centuries (Morigi Collection, Museo delle Culture, Lugano, Switzerland). The results facilitated a complete characterisation of the materials and demonstrated that the protocol can be used to study a wide variety of cellulosic materials, including both natural and man-modified fibres, and paper

Historical silk: a novel method to evaluate degumming with non-invasive infrared spectroscopy and spectral deconvolution

: To correctly manage a collection of historical silks, it is important to detect if the yarn has been originally subjected to degumming. This process is generally applied to eliminate sericin; the obtained fiber is named soft silk, in contrast with hard silk which is unprocessed. The distinction between hard and soft silk gives both historical information and useful indications for informed conservation. With this aim, 32 samples of silk textiles from traditional Japanese samurai armors (15th-20th century) were characterized in a non-invasive way. ATR-FTIR spectroscopy has been previously used to detect hard silk, but data interpretation is challenging. To overcome this difficulty, an innovative analytical protocol based on external reflection FTIR (ER-FTIR) spectroscopy was employed, coupled with spectral deconvolution and multivariate data analysis. The ER-FTIR technique is rapid, portable, and widely employed in the cultural heritage field, but rarely applied to the study of textiles. The ER-FTIR band assignment for silk was discussed for the first time. Then, the evaluation of the OH stretching signals allowed for a reliable distinction between hard and soft silk. Such an innovative point of view, which exploits a "weakness" of FTIR spectroscopy-the strong absorption from water molecules-to indirectly obtain the results, can have industrial applications too

Accurate and rapid antibiotic susceptibility testing using a machine learning-assisted nanomotion technology platform.

Antimicrobial resistance (AMR) is a major public health threat, reducing treatment options for infected patients. AMR is promoted by a lack of access to rapid antibiotic susceptibility tests (ASTs). Accelerated ASTs can identify effective antibiotics for treatment in a timely and informed manner. We describe a rapid growth-independent phenotypic AST that uses a nanomotion technology platform to measure bacterial vibrations. Machine learning techniques are applied to analyze a large dataset encompassing 2762 individual nanomotion recordings from 1180 spiked positive blood culture samples covering 364 Escherichia coli and Klebsiella pneumoniae isolates exposed to cephalosporins and fluoroquinolones. The training performances of the different classification models achieve between 90.5 and 100% accuracy. Independent testing of the AST on 223 strains, including in clinical setting, correctly predict susceptibility and resistance with accuracies between 89.5% and 98.9%. The study shows the potential of this nanomotion platform for future bacterial phenotype delineation