Large-scale analyses of the X chromosome in 2,354 infertile men discover recurrently affected genes associated with spermatogenic failure.

Although the evolutionary history of the X chromosome indicates its specialization in male fitness, its role in spermatogenesis has largely been unexplored. Currently only three X chromosome genes are considered of moderate-definitive diagnostic value. We aimed to provide a comprehensive analysis of all X chromosome-linked protein-coding genes in 2,354 azoospermic/cryptozoospermic men from four independent cohorts. Genomic data were analyzed and compared with data in normozoospermic control individuals and gnomAD. While updating the clinical significance of known genes, we propose 21 recurrently mutated genes strongly associated with and 34 moderately associated with azoospermia/cryptozoospermia not previously linked to male infertility (novel). The most frequently affected prioritized gene, RBBP7, was found mutated in ten men across all cohorts, and our functional studies in Drosophila support its role in germ stem cell maintenance. Collectively, our study represents a significant step towards the definition of the missing genetic etiology in idiopathic severe spermatogenic failure and significantly reduces the knowledge gap of X-linked genetic causes of azoospermia/cryptozoospermia contributing to the development of future diagnostic gene panels.

A Long-Term Comparison between the AethLabs MA350 and Aerosol Magee Scientific AE33 Black Carbon Monitors in the Greater Salt Lake City Metropolitan Area.

Black carbon (BC) or soot contains ultrafine combustion particles that are associated with a wide range of health impacts, leading to respiratory and cardiovascular diseases. Both long-term and short-term health impacts of BC have been documented, with even low-level exposures to BC resulting in negative health outcomes for vulnerable groups. Two aethalometers-AethLabs MA350 and Aerosol Magee Scientific AE33-were co-located at a Utah Division of Air Quality site in Bountiful, Utah for just under a year. The aethalometer comparison showed a close relationship between instruments for IR BC, Blue BC, and fossil fuel source-specific BC estimates. The biomass source-specific BC estimates were markedly different between instruments at the minute and hour scale but became more similar and perhaps less-affected by high-leverage outliers at the daily time scale. The greater inter-device difference for biomass BC may have been confounded by very low biomass-specific BC concentrations during the study period. These findings at a mountainous, high-elevation, Greater Salt Lake City Area site support previous study results and broaden the body of evidence validating the performance of the MA350.

Actionable secondary findings following exome sequencing of 836 non-obstructive azoospermia cases and their value in patient management.

STUDY QUESTION: What is the load, distribution and added clinical value of secondary findings (SFs) identified in exome sequencing (ES) of patients with non-obstructive azoospermia (NOA)? SUMMARY ANSWER: One in 28 NOA cases carried an identifiable, medically actionable SF. WHAT IS KNOWN ALREADY: In addition to molecular diagnostics, ES allows assessment of clinically actionable disease-related gene variants that are not connected to the patient's primary diagnosis, but the knowledge of which may allow the prevention, delay or amelioration of late-onset monogenic conditions. Data on SFs in specific clinical patient groups, including reproductive failure, are currently limited. STUDY DESIGN, SIZE, DURATION: The study group was a retrospective cohort of patients with NOA recruited in 10 clinics across six countries and formed in the framework of the international GEMINI (The GEnetics of Male INfertility Initiative) study. PARTICIPANTS/MATERIALS, SETTING, METHODS: ES data of 836 patients with NOA were exploited to analyze SFs in 85 genes recommended by the American College of Medical Genetics and Genomics (ACMG), Geisinger's MyCode, and Clinical Genome Resource. The identified 6374 exonic variants were annotated with ANNOVAR and filtered for allele frequency, retaining 1381 rare or novel missense and loss-of-function variants. After automatic assessment of pathogenicity with ClinVar and InterVar, 87 variants were manually curated. The final list of confident disease-causing SFs was communicated to the corresponding GEMINI centers. When patient consent had been given, available family health history and non-andrological medical data were retrospectively assessed. MAIN RESULTS AND THE ROLE OF CHANCE: We found a 3.6% total frequency of SFs, 3.3% from the 59 ACMG SF v2.0 genes. One in 70 patients carried SFs in genes linked to familial cancer syndromes, whereas 1 in 60 cases was predisposed to congenital heart disease or other cardiovascular conditions. Retrospective assessment confirmed clinico-molecular diagnoses in several cases. Notably, 37% (11/30) of patients with SFs carried variants in genes linked to male infertility in mice, suggesting that some SFs may have a co-contributing role in spermatogenic impairment. Further studies are needed to determine whether these observations represent chance findings or the profile of SFs in NOA patients is indeed different from the general population. LIMITATIONS, REASONS FOR CAUTION: One limitation of our cohort was the low proportion of non-Caucasian ethnicities (9%). Additionally, as comprehensive clinical data were not available retrospectively for all men with SFs, we were not able to confirm a clinico-molecular diagnosis and assess the penetrance of the specific variants. WIDER IMPLICATIONS OF THE FINDINGS: For the first time, this study analyzed medically actionable SFs in men with spermatogenic failure. With the evolving process to incorporate ES into routine andrology practice for molecular diagnostic purposes, additional assessment of SFs can inform about future significant health concerns for infertility patients. Timely detection of SFs and respective genetic counseling will broaden options for disease prevention and early treatment, as well as inform choices and opportunities regarding family planning. A notable fraction of SFs was detected in genes implicated in maintaining genome integrity, essential in both mitosis and meiosis. Thus, potential genetic pleiotropy may exist between certain adult-onset monogenic diseases and NOA. STUDY FUNDING/COMPETING INTEREST(S): This work was supported by the Estonian Research Council grants IUT34-12 and PRG1021 (M.L. and M.P.); National Institutes of Health of the United States of America grant R01HD078641 (D.F.C., K.I.A. and P.N.S.); National Institutes of Health of the United States of America grant P50HD096723 (D.F.C. and P.N.S.); National Health and Medical Research Council of Australia grant APP1120356 (M.K.O'B., D.F.C. and K.I.A.); Fundação para a Ciência e a Tecnologia (FCT)/Ministério da Ciência, Tecnologia e Inovação grant POCI-01-0145-FEDER-007274 (A.M.L., F.C. and J.G.) and FCT: IF/01262/2014 (A.M.L.). J.G. was partially funded by FCT/Ministério da Ciência, Tecnologia e Ensino Superior (MCTES), through the Centre for Toxicogenomics and Human Health-ToxOmics (grants UID/BIM/00009/2016 and UIDB/00009/2020). M.L.E. is a consultant for, and holds stock in, Roman, Sandstone, Dadi, Hannah, Underdog and has received funding from NIH/NICHD. Co-authors L.K., K.L., L.N., K.I.A., P.N.S., J.G., F.C., D.M.-M., K.A., K.A.J., M.K.O'B., A.M.L., D.F.C., M.P. and M.L. declare no conflict of interest. TRIAL REGISTRATION NUMBER: N/A.

Forecasting of a complex phenomenon using stochastic data-based techniques under non-conventional schemes: The SARS-CoV-2 virus spread case.

Epidemics are complex dynamical processes that are difficult to model. As revealed by the SARS-CoV-2 pandemic, the social behavior and policy decisions contribute to the rapidly changing behavior of the virus' spread during outbreaks and recessions. In practice, reliable forecasting estimations are needed, especially during early contagion stages when knowledge and data are insipient. When stochastic models are used to address the problem, it is necessary to consider new modeling strategies. Such strategies should aim to predict the different contagious phases and fast changes between recessions and outbreaks. At the same time, it is desirable to take advantage of existing modeling frameworks, knowledge and tools. In that line, we take Autoregressive models with exogenous variables (ARX) and Vector autoregressive (VAR) techniques as a basis. We then consider analogies with epidemic's differential equations to define the structure of the models. To predict recessions and outbreaks, the possibility of updating the model's parameters and stochastic structures is considered, providing non-stationarity properties and flexibility for accommodating the incoming data to the models. The Generalized-Random-Walk (GRW) and the State-Dependent-Parameter (SDP) techniques shape the parameters' variability. The stochastic structures are identified following the Akaike (AIC) criterion. The models use the daily rates of infected, death, and healed individuals, which are the most common and accurate data retrieved in the early stages. Additionally, different experiments aim to explore the individual and complementary role of these variables. The results show that although both the ARX-based and VAR-based techniques have good statistical accuracy for seven-day ahead predictions, some ARX models can anticipate outbreaks and recessions. We argue that short-time predictions for complex problems could be attained through stochastic models that mimic the fundamentals of dynamic equations, updating their parameters and structures according to incoming data.

Steerable Surgical Instrument for Conventional and Single-Site Minimally Invasive Surgery.

Background. This article aims to present an innovative design of a steerable surgical instrument for conventional and single-site minimally invasive surgery (MIS), which improves the dexterity and maneuverability of the surgeon while offering a solution to the limitations of current tools. Methods. The steerable MIS instrument consists of a deflection structure with a curved sliding joints design that articulates the distal tip in two additional degrees of freedom (DoFs), relative to the instrument shaft, using transmission by cables. A passive ball-joint mechanism articulates the handle relative to the instrument shaft, improves wrist posture, and prevents collision of instrument handles during single-site MIS procedures. The two additional DoFs of the articulating tip are activated by a thumb-controlled device, using a joystick design mounted on the handle. This steerable MIS instrument was developed by additive manufacturing in a 3D printer using PLA polymer. Results. Prototype testing showed a maximum tip deflection of 60° in the left and right directions, with a total deflection of 120°. With the passive ball-joint fully offset, the steerable tip achieved a deflection of 90° for the right and 40° for the left direction, with a total deflection of 130°. Furthermore, the passive ball-joint mechanism in the handle obtained a maximum range of motion of 60°. Conclusions. This steerable MIS instrument concept offers an alternative to enhance the application fields of conventional and single-site MIS, increasing manual dexterity of the surgeon and the ability to reach narrow anatomies from other directions.

Intra-city variability of fine particulate matter during COVID-19 lockdown: A case study from Park City, Utah.

Urban air quality is a growing concern due a range of social, economic, and health impacts. Since the SARS-CoV-19 pandemic began in 2020, governments have produced a range of non-medical interventions (NMIs) (e.g. lockdowns, stay-at-home orders, mask mandates) to prevent the spread of COVID-19. A co-benefit of NMI implementation has been the measurable improvement in air quality in cities around the world. Using the lockdown policy of the COVID-19 pandemic as a natural experiment, we traced the changing emissions patterns produced under the pandemic in a mid-sized, high-altitude city to isolate the effects of human behavior on air pollution. We tracked air pollution over time periods reflecting the Pre-Lockdown, Lockdown, and Reopening stages, using high quality, research grade sensors in both commercial and residential areas to better understand how each setting may be uniquely impacted by pollution downturn events. Based on this approach, we found the commercial area of the city showed a greater decrease in air pollution than residential areas during the lockdown period, while both areas experienced a similar rebound post lockdown. The easing period following the lockdown did not lead to an immediate rebound in human activity and the air pollution increase associated with reopening, took place nearly two months after the lockdown period ended. We hypothesize that differences in heating needs, travel demands, and commercial activity, are responsible for the corresponding observed changes in the spatial distribution of pollutants over the study period. This research has implications for climate policy, low-carbon energy transitions, and may even impact local policy due to changing patterns in human exposure that could lead to important public health outcomes, if left unaddressed.

Long-term urban carbon dioxide observations reveal spatial and temporal dynamics related to urban characteristics and growth.

Cities are concentrated areas of CO2 emissions and have become the foci of policies for mitigation actions. However, atmospheric measurement networks suitable for evaluating urban emissions over time are scarce. Here we present a unique long-term (decadal) record of CO2 mole fractions from five sites across Utah's metropolitan Salt Lake Valley. We examine "excess" CO2 above background conditions resulting from local emissions and meteorological conditions. We ascribe CO2 trends to changes in emissions, since we did not find long-term trends in atmospheric mixing proxies. Three contrasting CO2 trends emerged across urban types: negative trends at a residential-industrial site, positive trends at a site surrounded by rapid suburban growth, and relatively constant CO2 over time at multiple sites in the established, residential, and commercial urban core. Analysis of population within the atmospheric footprints of the different sites reveals approximately equal increases in population influencing the observed CO2, implying a nonlinear relationship with CO2 emissions: Population growth in rural areas that experienced suburban development was associated with increasing emissions while population growth in the developed urban core was associated with stable emissions. Four state-of-the-art global-scale emission inventories also have a nonlinear relationship with population density across the city; however, in contrast to our observations, they all have nearly constant emissions over time. Our results indicate that decadal scale changes in urban CO2 emissions are detectable through monitoring networks and constitute a valuable approach to evaluate emission inventories and studies of urban carbon cycles.

Changes in the physiological and biochemical state of peanut plants (Arachis hypogaea L.) induced by exposure to green metallic nanoparticles.

Different types of nanoparticles (NPs) are increasingly used in multiple sectors such as industry, medicine and agriculture. This application has increased the possibility of NPs accumulating and contaminating the environment. Plants are one of the essential building blocks of all ecosystems and the interaction between NPs and plants is an indispensable aspect of risk assessment. To understand the effects of NPs in agricultural systems, in the present study we investigated the effects of exposure of Ag, Cu and Cu/Ag phytonanoparticles in Arachis hypogaea L. plants at a physiological and biochemical level, for which NPs solutions were applied foliarly at concentrations of 250, 500, 750 and 1000 ppm for 48 days. Parameters such as leaf length, chlorophyll and concentration of phytohormones showed that phytonanoparticles could cause serious damage to plant growth and development. Plants exposed to phytonanoparticles showed an increase in total phenols, proline, PAL activity and antioxidant enzymes, this to mitigate the stress caused. The alteration in the composition and content of fatty acids in the peanut kernels after exposure to different NPs indicated that they could affect the yield and quality of crop. Therefore, it is necessary to investigate its potential impact on food quality. Statement of noveltyIn this manuscript, we report for the first time that green nanoparticles induced a lower degree of toxicity in plants compared to commercial nanoparticles.Our results indicate that the mechanisms by which peanut plants respond to the application of nanoparticles were an increase in the activity of phenylalanine ammonia-lyase and antioxidant enzymes. So far there are few studies on the effect of nanoparticles on plant hormones, our results revealed a significant decrease in indole-3 acetic acid and induced the synthesis of gibberellins. The modification in the composition and content of fatty acids in the peanut kernels indicated that the nanoparticles could affect the quality of the crop.

Zinc Oxide Phytonanoparticles' Effects on Yield and Mineral Contents in Fruits of Tomato (Solanum lycopersicum L. cv. Cherry) under Field Conditions.

The use of phytonanoparticles in agriculture could decrease the use of fertilizers and therefore decrease soil contamination, due to their size being better assimilated in plants. It is important to mention that the nanofertilizer is slow-releasing and improves plant physiological properties and various nutritional parameters. The influence of soil and foliar applications of phytonanoparticles of ZnO with the Moringa oleifera extract under three concentrations (25, 50, and 100 ppm) was evaluated on the cherry tomato crop (Solanum lycopersicum L.). Synthesis of the phytonanoparticles was analyzed with ultraviolet-visible spectroscopy (UV-Vis) and infrared transmission spectroscopy with Fourier transform (FT-IR), as well as the analysis with the dynamic light scattering (DLS) technique. The morphometric parameters were evaluated before and after the application of the nanoparticles. The minerals' content of fruits was done 95 days after planting. Results showed that soil application was better at a concentration of 25 ppm of phytonanoparticles since it allowed the greatest number of flowers and fruits on the plant; however, it was demonstrated that when performing a foliar application, the fruit showed the highest concentrations for the elements Mg, Ca, and Na at concentrations of 511, 4589, and 223 mg kg-1, respectively.

Genetic dissection of spermatogenic arrest through exome analysis: clinical implications for the management of azoospermic men.

PURPOSE: Azoospermia affects 1% of men and it can be the consequence of spermatogenic maturation arrest (MA). Although the etiology of MA is likely to be of genetic origin, only 13 genes have been reported as recurrent potential causes of MA. METHODS: Exome sequencing in 147 selected MA patients (discovery cohort and two validation cohorts). RESULTS: We found strong evidence for five novel genes likely responsible for MA (ADAD2, TERB1, SHOC1, MSH4, and RAD21L1), for which mouse knockout (KO) models are concordant with the human phenotype. Four of them were validated in the two independent MA cohorts. In addition, nine patients carried pathogenic variants in seven previously reported genes-TEX14, DMRT1, TEX11, SYCE1, MEIOB, MEI1, and STAG3-allowing to upgrade the clinical significance of these genes for diagnostic purposes. Our meiotic studies provide novel insight into the functional consequences of the variants, supporting their pathogenic role. CONCLUSION: Our findings contribute substantially to the development of a pre-testicular sperm extraction (TESE) prognostic gene panel. If properly validated, the genetic diagnosis of complete MA prior to surgical interventions is clinically relevant. Wider implications include the understanding of potential genetic links between nonobstructive azoospermia (NOA) and cancer predisposition, and between NOA and premature ovarian failure.

Constraining Urban CO2 Emissions Using Mobile Observations from a Light Rail Public Transit Platform.

Urban environments are characterized by pronounced spatiotemporal heterogeneity, which can present sampling challenges when utilizing conventional greenhouse gas (GHG) measurement systems. In Salt Lake City, Utah, a GHG instrument was deployed on a light rail train car that continuously traverses the Salt Lake Valley (SLV) through a range of urban typologies. CO2 measurements from a light rail train car were used within a Bayesian inverse modeling framework to constrain urban emissions across the SLV during the fall of 2015. The primary objectives of this study were to (1) evaluate whether ground-based mobile measurements could be used to constrain urban emissions using an inverse modeling framework and (2) quantify the information that mobile observations provided relative to conventional GHG monitoring networks. Preliminary results suggest that ingesting mobile measurements into an inverse modeling framework generated a posterior emission estimate that more closely aligned with observations, reduced posterior emission uncertainties, and extends the geographical extent of emission adjustments.

A partner-switching system controls activation of mixed-linkage ß-glucan synthesis by c-di-GMP in Sinorhizobium meliloti.

Sinorhizobium meliloti synthesizes a linear mixed-linkage (1 → 3)(1 → 4)-ß-d-glucan (ML ß-glucan, MLG) in response to high levels of cyclic diguanylate (c-di-GMP). Two proteins BgsA and BgsB are required for MLG synthesis, BgsA being the glucan synthase which is activated upon c-di-GMP binding to its C-terminal domain. Here we report that the product of bgrR (SMb20447) is a diguanylate cyclase (DGC) that provides c-di-GMP for the synthesis of MLG by BgsA. bgrR is the first gene of a hexacistronic bgrRSTUWV operon, likely encoding a partner-switching regulatory network where BgrR is the final target. Using different approaches, we have determined that the products of genes bgrU (containing a putative PP2C serine phosphatase domain) and bgrW (with predicted kinase effector domain), modulate the phosphorylation status and the activity of the STAS domain protein BgrV. We propose that unphosphorylated BgrV inhibits BgrR DGC activity, perhaps through direct protein-protein interactions as established for other partner switchers. A bgrRSTUWV operon coexists with MLG structural bgsBA genes in many rhizobial genomes but is also present in some MLG non-producers, suggesting a role of this partner-switching system in other processes besides MLG biosynthesis.

From exome analysis in idiopathic azoospermia to the identification of a high-risk subgroup for occult Fanconi anemia.

PURPOSE: In about 10% of patients affected by Fanconi anemia (FA) the diagnosis is delayed until adulthood, and the presenting symptom in these "occult" FA cases is often a solid cancer and cancer treatment-related toxicity. Highly predictive clinical parameter(s) for diagnosing such an adult-onset cases are missing. METHODS: (1) Exome sequencing (ES), (2) Sanger sequencing of FANCA, (3) diepoxybutane (DEB)-induced chromosome breakage test. RESULTS: ES identified a pathogenic homozygous FANCA variant in a patient affected by Sertoli cell-only syndrome (SCOS) and in his azoospermic brother. Although they had no overt anemia, chromosomal breakage test revealed a reverse somatic mosaicism in the former and a typical FA picture in the latter. In 27 selected SCOS cases, 1 additional patient showing compound heterozygous pathogenic FANCA variants was identified with positive chromosomal breakage test. CONCLUSION: We report an extraordinarily high frequency of FA in a specific subgroup of azoospermic patients (7.1%). The screening for FANCA pathogenic variants in such patients has the potential to identify undiagnosed FA before the appearance of other severe clinical manifestations of the disease. The definition of this high-risk group for "occult" FA, based on specific testis phenotype with mild/borderline hematological alterations, is of unforeseen clinical relevance.

Silver nanoparticles from Justicia spicigera and their antimicrobial potentialities in the biocontrol of foodborne bacteria and phytopathogenic fungi.

In the present work, the biosynthesis of silver-nanoparticles (AgNP) was evaluated using the aqueous extract from Justicia spicigera. The obtained silver nanoparticles were characterized using UV-visible spectroscopic techniques, energy dispersive X-ray spectrometers (EDS), zeta potential and dynamic light scattering. The antimicrobial activity of biosynthesized AgNP was tested against foodborne bacteria (Bacillus cereus, Klebsiella pneumoniae and Enterobacter aerogenes) and phytopathogenic fungi (Colletotrichum sp., Fusarium solani, Alternaria alternata and Macrophomina phaseolina). The elemental profile of synthesized nanoparticles using J. spicigera shows higher counts at 3keV due to silver, confirming the formation of silver nanoparticles. Scanning electron microscopy (SEM) analysis showed a particle size between 86 and 100nm with spherical morphology. AgNP showed effective antibacterial and antifungal activity against the tested organisms principally with B. cereus, K. pneumoniae, E. aerogenes, A. alternata and M. phaseolina. Therefore, further studies are needed to confirm the potential of AgNP from J. spicigera in the control of indicator organisms under field conditions.

Social factors and aromatase gene expression during adult male-to-female sex change in captive leopard grouper Mycteroperca rosacea.

Social factors and aromatase gene expression in the leopard grouper Mycteroperca rosacea was studied when captive fish were separated by sex during the reproductive (April-June) and post-reproductive (July-September) seasons. Monosex females, monosex males, and mixed-sex, held in social sextet units were analyzed for sex steroids throughout confinement. At the end of the experiment, the gonad-sex was defined by histology, and gonad and brain aromatase gene expressions were quantified. Only males held in the monosex social units changed sex. Histology showed one male remained unchanged, six were found in a transitional sexual stage, in which two had intersex-predominantly-testes, and four had a more defined intersex ovo-testes pattern, and 11 were immature de novo females (neofemales). Neofemales and most intersex fish did not survive. In spring, 11-ketosterone showed a specific male profile, which suggests that male-to-female sex change was not triggered during the reproductive season. The low steroid levels in summer made it impossible to associate the sex change to a gonad hormonal shift; in September, gonad aromatase gene expression was not significantly different among groups. However, brain aromatase expression in intersex fish was significantly higher than monosex females, mixed-sex females, and neofemale groups. These results suggest that in the absence of female hormonal compounds, and at a time when male gonad steroidogenesis was diminished, the brain mediated male-to-male social-behavioral interactions, including stress, by increasing aromatization, resulting in derived intersex-male, which triggered more aromatization, followed by a sex change.

Novel mixed-linkage ß-glucan activated by c-di-GMP in Sinorhizobium meliloti.

An artificial increase of cyclic diguanylate (c-di-GMP) levels in Sinorhizobium meliloti 8530, a bacterium that does not carry known cellulose synthesis genes, leads to overproduction of a substance that binds the dyes Congo red and calcofluor. Sugar composition and methylation analyses and NMR studies identified this compound as a linear mixed-linkage (1 → 3)(1 → 4)-ß-D-glucan (ML ß-glucan), not previously described in bacteria but resembling ML ß-glucans found in plants and lichens. This unique polymer is hydrolyzed by the specific endoglucanase lichenase, but, unlike lichenan and barley glucan, it generates a disaccharidic → 4)-ß-D-Glcp-(1 → 3)-ß-D-Glcp-(1 → repeating unit. A two-gene operon bgsBA required for production of this ML ß-glucan is conserved among several genera within the order Rhizobiales, where bgsA encodes a glycosyl transferase with domain resemblance and phylogenetic relationship to curdlan synthases and to bacterial cellulose synthases. ML ß-glucan synthesis is subjected to both transcriptional and posttranslational regulation. bgsBA transcription is dependent on the exopolysaccharide/quorum sensing ExpR/SinI regulatory system, and posttranslational regulation seems to involve allosteric activation of the ML ß-glucan synthase BgsA by c-di-GMP binding to its C-terminal domain. To our knowledge, this is the first report on a linear mixed-linkage (1 → 3)(1 → 4)-ß-glucan produced by a bacterium. The S. meliloti ML ß-glucan participates in bacterial aggregation and biofilm formation and is required for efficient attachment to the roots of a host plant, resembling the biological role of cellulose in other bacteria.

Changes in the phenylalanine ammonia lyase activity, total phenolic compounds, and flavonoids in Prosopis glandulosa treated with cadmium and copper.

The aim of the present work is to evaluate the changes on the phenylalanine ammonia lyase (PAL) activity, phenolic compounds accumulation and photochemical efficiency in leaves of P. glandulosa treated with Cd2+ (0.001 M) and Cu2+ (0.52 M) concentrations for 96 h under hydroponic conditions. The results showed that only leaves treated with copper had a decrease in photochemical efficiency and leaf epidermal polyphenols in P. glandulosa leaves after 96 h of exposure. On the other hand the reverse-phase HPLC analysis revealed higher levels of phenolic compound (gallic, vanillic and caffeic acids) and flavonoids (rutin and kaempferol-3-O-glucosides) in plant leaves from Cu and Cd-treatments with respect to control plants. Finally, highest increments in PAL activity was observed in extracts of leaves treated with Cu and Cd (about 205 and 284%), respectively, with respect to control plants after 96 h treatment. These suggest that activation of phenylpropanoid pathway represent a source of nonenzymatic antioxidants that protect at P. glandulosa against oxidative stress when exposed to cadmium and copper. Hence future studies are necessary to elucidate the participation of phenylpropanoid pathway in the reduction of metal toxicity in Prosopis species.

Pumilacidins from the Octocoral-Associated Bacillus sp. DT001 Display Anti-Proliferative Effects in Plasmodium falciparum.

Chemical examination of the octocoral-associated Bacillus species (sp.) DT001 led to the isolation of pumilacidins A (1) and C (2). We investigated the effect of these compounds on the viability of Plasmodium falciparum and the mechanism of pumilacidin-induced death. The use of inhibitors of protein kinase C (PKC) and phosphoinositide 3-kinase (PI3K) was able to prevent the effects of pumilacidins A and C. The results indicated also that pumilacidins inhibit parasite growth via mitochondrial dysfunction and decreased cytosolic Ca2+.

AmrZ regulates cellulose production in Pseudomonas syringae pv. tomato DC3000.

In Pseudomonas syringae pv. tomato DC3000, the second messenger c-di-GMP has been previously shown to stimulate pellicle formation and cellulose biosynthesis. A screen for genes involved in cellulose production under high c-di-GMP intracellular levels led to the identification of insertions in two genes, wssB and wssE, belonging to the Pto DC3000 cellulose biosynthesis operon wssABCDEFGHI. Interestingly, beside cellulose-deficient mutants, colonies with a rougher appearance than the wild type also arouse among the transposants. Those mutants carry insertions in amrZ, a gene encoding a transcriptional regulator in different Pseudomonas. Here, we provide evidence that AmrZ is involved in the regulation of bacterial cellulose production at transcriptional level by binding to the promoter region of the wssABCDEFGHI operon and repressing cellulose biosynthesis genes. Mutation of amrZ promotes wrinkly colony morphology, increased cellulose production and loss of motility in Pto DC3000. AmrZ regulon includes putative c-di-GMP metabolising proteins, like AdcA and MorA, which may also impact those phenotypes. Furthermore, an amrZ but not a cellulose-deficient mutant turned out to be impaired in pathogenesis, indicating that AmrZ is a key regulator of Pto DC3000 virulence probably by controlling bacterial processes other than cellulose production.

Novel hematological parameters for the evaluation of patients with myeloproliferative neoplasms: the immature platelet and reticulocyte fractions.

New automated hematology analyzers have led to the availability of novel hematological parameters, including the immature platelet fraction (IPF) and the immature reticulocyte fraction (IRF), both of potential interest in patients with myeloproliferative neoplasms (MPNs). We performed a prospective analysis of 217 patients with MPN, including 32 (15%) with essential thrombocythemia (ET), 43 (20%) with polycythemia vera (PV), and 142 (65%) with myelofibrosis (MF); the IPF and IRF were measured by the Sysmex XN analyzer. As compared to patients with ET, both a higher IPF and IRF were observed among patients with PV and MF. Factors associated with high IPF among patients with PV/ET were male sex, thrombocytopenia, and diagnosis of PV; among patients with MF, they were elevated peripheral blasts, low platelet count, JAK2 V617F mutation, and previous therapy. Factors associated with high IRF among patients with PV/ET were low hemoglobin, high reticulocyte count, and PV diagnosis; among patients with MF, they were peripheral blasts and elevated reticulocytes. The IPF and IRF represent novel parameters in patients with MPN with potential relevant clinical implications. Comparison with healthy subjects and those with secondary polycythemia is needed to confirm our preliminary findings.