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NANOS3 is expressed in migrating primordial germ cells (PGCs) to protect them from apoptosis, and it is known to be a critical factor for germline development of both sexes in several organisms. However, to date, live NANOS3 knockout (KO) cattle have not been reported, and the specific role of NANOS3 in male cattle, or bulls, remains unexplored. This study generated NANOS3 KO cattle via cytoplasmic microinjection of the CRISPR/Cas9 system in vitro produced bovine zygotes and evaluated the effect of NANOS3 elimination on bovine germline development, from fetal development through reproductive age. The co-injection of two selected guide RNA (gRNA)/Cas9 ribonucleoprotein complexes (i.e., dual gRNA approach) at 6 h post fertilization achieved a high NANOS3 KO rate in developing embryos. Subsequent embryo transfers resulted in a 31% (n = 8/26) pregnancy rate. A 75% (n = 6/8) total KO rate (i.e., 100% of alleles present contained complete loss-of-function mutations) was achieved with the dual gRNA editing approach. In NANOS3 KO fetal testes, PGCs were found to be completely eliminated by 41-day of fetal age. Importantly, despite the absence of germ cells, seminiferous tubule development was not impaired in NANOS3 KO bovine testes during fetal, perinatal, and adult stages. Moreover, a live, NANOS3 KO, germline-ablated bull was produced and at sexual maturity he exhibited normal libido, an anatomically normal reproductive tract, and intact somatic gonadal development and structure. Additionally, a live, NANOS3 KO, germline-ablated heifer was produced. However, it was evident that the absence of germ cells in NANOS3 KO cattle compromised the normalcy of ovarian development to a greater extent than it did testes development. The meat composition of NANOS3 KO cattle was unremarkable. Overall, this study demonstrated that the absence of NANOS3 in cattle leads to the specific deficiency of both male and female germ cells, suggesting the potential of NANOS3 KO cattle to act as hosts for donor-derived exogenous germ cell production in both sexes. These findings contribute to the understanding of NANOS3 function in cattle and have valuable implications for the development of novel breeding technologies using germline complementation in NANOS3 KO germline-ablated hosts.

ABSTRACT Facultative marine bacterial pathogens sense environmental signals so that the expression of virulence factors is upregulated on entry into hosts and downregulated during the free-living lifestyle in the environment. In this study, we utilized transcriptome sequencing to compare the transcriptional profiles of Photobacterium damselae subsp. damselae, a generalist pathogen that causes disease in diverse marine animals and fatal infections in humans at NaCl concentrations that mimic the free-living lifestyle or host internal milieu, respectively. We here show that NaCl concentration constitutes a major regulatory signal that shapes the transcriptome and uncover 1,808 differentially expressed genes (888 upregulated and 920 downregulated in response to low-salt conditions). Growth at 3% NaCl, a salinity that mimics the free-living lifestyle, upregulated genes involved in energy production, nitrogen metabolism, transport of compatible solutes, utilization of trehalose and fructose, and carbohydrate and amino acid metabolism with strong upregulation of the arginine deiminase system (ADS). In addition, we observed a marked increase in resistance to antibiotics at 3% NaCl. On the contrary, the low salinity conditions (1% NaCl) that mimic those encountered in the host triggered a virulence gene expression profile that maximized the production of the type 2 secretion system (T2SS)-dependent cytotoxins damselysin, phobalysin P, and a putative PirAB-like toxin, observations that were corroborated by the analysis of the secretome. Low salinity also upregulated the expression of iron-acquisition systems, efflux pumps, and other functions related to stress response and virulence. The results of this study greatly expand our knowledge of the salinity-responsive adaptations of a generalist and versatile marine pathogen. IMPORTANCE Pathogenic Vibrionaceae species experience continuous shifts of NaCl concentration in their life cycles. However, the impact of salinity changes in gene regulation has been studied in a small number of Vibrio species. In this study, we analyzed the transcriptional response of Photobacterium damselae subsp. damselae (Pdd), a generalist and facultative pathogen, to changes in salinity, and demonstrate that growth at 1% NaCl in comparison to 3% NaCl triggers a virulence program of gene expression, with a major impact in the T2SS-dependent secretome. The decrease in NaCl concentration encountered by bacteria on entry into a host is proposed to constitute a regulatory signal that upregulates a genetic program involved in host invasion and tissue damage, nutrient scavenging (notably iron), and stress responses. This study will surely inspire new research on Pdd pathobiology, as well as on other important pathogens of the family Vibrionaceae and related taxa whose salinity regulons still await investigation.

The progress made in recent years in the derivation and culture of pluripotent stem cells from farm animals opens up the possibility of creating livestock chimeras. Chimeras producing gametes exclusively derived from elite donor stem cells could pass superior genetics on to the next generation and thereby reduce the genetic lag that typically exists between the elite breeding sector and the commercial production sector, especially for industries like beef and sheep where genetics is commonly disseminated through natural service mating. Chimeras carrying germ cells generated from genome-edited or genetically engineered pluripotent stem cells could further disseminate useful genomic alterations such as climate adaptation, animal welfare improvements, the repair of deleterious genetic conditions, and/or the elimination of undesired traits such as disease susceptibility to the next generation. Despite the successful production of chimeras with germ cells generated from pluripotent donor stem cells injected into preimplantation-stage blastocysts in model species, there are no documented cases of this occurring in livestock. Here, we review the literature on the derivation of pluripotent stem cells from ungulates, and progress in the production of chimeric ungulate livestock for agricultural applications, drawing on insights from studies done in model species, and discuss future possibilities of this fast-moving and developing field. Aside from the technical aspects, the consistency of the regulatory approach taken by different jurisdictions towards chimeric ungulate livestock with germ cells generated from pluripotent stem cells and their progeny will be an important determinant of breeding industry uptake and adoption in animal agriculture.

ABSTRACT Stepped spillways can dissipate a great amount of energy during the flow passage over the chute, however these structures have limited operation due to the risk of cavitation damage. The induced aeration may protect the concrete chute through the air concentration near the channel bottom. Furthermore, some research studies have indicated that the presence of air in flows may reduce the mean pressures. The present research aims to analyze mean pressures, air entrainment coefficient and flow behavior over a stepped spillway with aeration induced by two different deflectors, comparing the results to natural aeration flow. Despite the jet impact influence, the induced aeration does not change significantly the mean pressures compared to natural aeration flow. The air entrainment coefficient, as well as the jet impact position, is higher for the deflector with the longer extension and, although air bubbles can be seen throughout the extension of the chute due to the air entrainment through the inferior flow surface, the induced aeration did not anticipate the boundary layer inception point position.

Peptidoglycan (PG) is a major component of the bacterial cell wall formed by long chains of two alternating sugars interconnected by short peptides, generating a mesh-like structure that enwraps the bacterial cell. Although PG provides structural integrity and support for anchoring other components of the cell envelope, it is constantly being remodeled through the action of specific enzymes that cleave or join its components.

The RstB histidine kinase of the two component system RstAB positively regulates the expression of damselysin (Dly), phobalysin P (PhlyP) and phobalysin C (PhlyC) cytotoxins in the fish and human pathogen Photobacterium damselae subsp. damselae, a marine bacterium of the family Vibrionaceae. However, the function of the predicted cognate response regulator RstA has not been studied so far, and the role of the RstAB system in other cell functions and phenotypes remain uninvestigated. Here, we analyzed the effect of rstA and rstB mutations in cell fitness and in diverse virulence-related features. Both rstA and rstB mutants were severely impaired in virulence for sea bream and sea bass fish. Mutants in rstA and rstB genes were impaired in hemolysis and in Dly-dependent phospholipase activity but had intact PlpV-dependent phospholipase and ColP-dependent gelatinase activities. rstA and rstB mutants grown at 0.5% NaCl exhibited impaired swimming motility, enlarged cell size and impaired ability to separate after cell division, whereas at 1% NaCl the mutants exhibited normal phenotypes. Mutation of any of the two genes also impacted tolerance to benzylpenicillin. Notably, rstA and rstB mutants showed impaired secretion of a number of type II secretion system (T2SS)-dependent proteins, which included the three major cytotoxins Dly, PhlyP and PhlyC, as well as a putative delta-endotoxin and three additional uncharacterized proteins which might constitute novel virulence factors of this pathogenic bacterium. The analysis of the T2SS-dependent secretome of P. damselae subsp. damselae also led to the identification of RstAB-independent potential virulence factors as lipoproteins, sialidases and proteases. The RstAB regulon included plasmid, chromosome I and chromosome II-encoded genes that showed a differential distribution among isolates of this subspecies. This study establishes RstAB as a major regulator of virulence and diverse cellular functions in P. damselae subsp. damselae.

The drastic reduction of agrobiodiversity has induced all countries in the world to implement numerous initiatives to safeguard genetic resources. The biodiversity issue is also one of the priority objectives of the global action plan for the conservation and sustainable use of FAO's cytogenetic resources and of the International Treaty on Plant Genetic Resources for Food and Agriculture. In Italy, the primary competence on genetic resources of agricultural interest is entrusted to the Ministry of Agricultural. In 2004, the Ministry entrusted the coordination of the RGV/FAO Project to CREA. The main objectives of the project, in addition to the inventory of accessions present in the collections of the structures involved, are their morphological, agronomic and molecular characterization and the productive enhancement with particular reference to the autochthonous varieties at risk of extinction. The project in question includes the activities of recovery and characterization of the viticultural germplasm illustrated in the present work. The 12 vines described here, like others yet to be rediscovered, are strongly linked to small winegrowers and indissolubly characterial of a territory in Campania Region that from the socio-cultural (rather fragmented agricultural system) and environmental orography point of view, like the national one, is very diversified.

A informação correta é determinante na qualidade do uso de medicamentos e na tomada de decisões em saúde. Assim, é fundamental a formação de profissionais capazes de interpretar a evidência científica sobre medicamentos, transformando-a em informação de fácil acesso. Nesse sentido, o Centro de Estudos do Medicamento (CEMED) da Faculdade de Farmácia da Universidade Federal de Minas Gerais (UFMG) promove a formação voltada para o uso racional de medicamentos, utilizando a divulgação de informações por meio de weblog (Blog do CEMED). A publicação no weblog depende do papel ativo dos discentes, os quais devem identificar as fontes de informação, selecionar os temas e elaborar, corrigir e divulgar as matérias. O blog tem registrado um movimento intenso de consulta, impulsionado pela interatividade das redes sociais. A iniciativa demonstra a evidente demanda por informações na área e se apresenta como importante recurso pedagógico, favorecendo o desenvolvimento de competências como avaliação crítica da literatura, comunicação escrita e trabalho em equipe.

NANOS3 is expressed in migrating primordial germ cells (PGCs) to protect them from apoptosis, and it is known to be a critical factor for germline development of both sexes in several organisms. However, to date, live NANOS3 knockout (KO) cattle have not been reported, and the specific role of NANOS3 in male cattle, or bulls, remains unexplored. This study generated NANOS3 KO cattle via cytoplasmic microinjection of the CRISPR/Cas9 system in vitro produced bovine zygotes and evaluated the effect of NANOS3 elimination on bovine germline development, from fetal development through reproductive age. The co-injection of two selected guide RNA (gRNA)/Cas9 ribonucleoprotein complexes (i.e., dual gRNA approach) at 6 h post fertilization achieved a high NANOS3 KO rate in developing embryos. Subsequent embryo transfers resulted in a 31% (n = 8/26) pregnancy rate. A 75% (n = 6/8) total KO rate (i.e., 100% of alleles present contained complete loss-of-function mutations) was achieved with the dual gRNA editing approach. In NANOS3 KO fetal testes, PGCs were found to be completely eliminated by 41-day of fetal age. Importantly, despite the absence of germ cells, seminiferous tubule development was not impaired in NANOS3 KO bovine testes during fetal, perinatal, and adult stages. Moreover, a live, NANOS3 KO, germline-ablated bull was produced and at sexual maturity he exhibited normal libido, an anatomically normal reproductive tract, and intact somatic gonadal development and structure. Additionally, a live, NANOS3 KO, germline-ablated heifer was produced. However, it was evident that the absence of germ cells in NANOS3 KO cattle compromised the normalcy of ovarian development to a greater extent than it did testes development. The meat composition of NANOS3 KO cattle was unremarkable. Overall, this study demonstrated that the absence of NANOS3 in cattle leads to the specific deficiency of both male and female germ cells, suggesting the potential of NANOS3 KO cattle to act as hosts for donor-derived exogenous germ cell production in both sexes. These findings contribute to the understanding of NANOS3 function in cattle and have valuable implications for the development of novel breeding technologies using germline complementation in NANOS3 KO germline-ablated hosts.

The origin of obscuration in active galactic nuclei (AGNs) is still an open debate. In particular, it is unclear what drives the relative contributions to the line-of-sight column densities from galaxy-scale and torus-linked obscuration. The latter source is expected to play a significant role in Unification Models, while the former is thought to be relevant in both Unification and Evolutionary models. In this work, we make use of a combination of cosmological semi-analytic models and semi-empirical prescriptions for the properties of galaxies and AGN, to study AGN obscuration. We consider a detailed object-by-object modelling of AGN evolution, including different AGN light curves (LCs), gas density profiles, and also AGN feedback-induced gas cavities. Irrespective of our assumptions on specific AGN LC or galaxy gas fractions, we find that, on the strict assumption of an exponential profile for the gas component, galaxy-scale obscuration alone can hardly reproduce the fraction of log (N H/cm−2) ≥ 24 sources at least at z ≲ 3. This requires an additional torus component with a thickness that decreases with luminosity to match the data. The torus should be present in all evolutionary stages of a visible AGN to be effective, although galaxy-scale gas obscuration may be sufficient to reproduce the obscured fraction with 22 < log (N H/cm−2) < 24 (Compton-thin, CTN) if we assume extremely compact gas disc components. The claimed drop of CTN fractions with increasing luminosity does not appear to be a consequence of AGN feedback, but rather of gas reservoirs becoming more compact with decreasing stellar mass. [ABSTRACT FROM AUTHOR]

Varietal identification plays a pivotal role in viticulture for several purposes. Nowadays, such identification is accomplished using ampelography and molecular markers, techniques requiring specific expertise and equipment. Deep learning, on the other hand, appears to be a viable and cost-effective alternative, as several recent studies claim that computer vision models can identify different vine varieties with high accuracy. Such works, however, limit their scope to a handful of selected varieties and do not provide accurate figures for external data validation. In the current study, five well-known computer vision models were applied to leaf images to verify whether the results presented in the literature can be replicated over a larger data set consisting of 27 varieties with 26 382 images. It was built over 2 years of dedicated field sampling at three geographically distinct sites, and a validation data set was collected from the Internet. Cross-validation results on the purpose-built data set confirm literature results. However, the same models, when validated against the independent data set, appear unable to generalize over the training data and retain the performances measured during cross validation. These results indicate that further enhancement have been done in filling such a gap and developing a more reliable model to discriminate among grape varieties, underlining that, to achieve this purpose, the image resolution appears to be a crucial factor in the development of such models. [ABSTRACT FROM AUTHOR]

In the standard quasar model, the accretion disc obscuration is due to the canonical dusty torus. Here, we argue that a substantial part of the quasar obscuration can come from the interstellar medium (ISM) when the quasars are embedded in compact starbursts. We use an obscuration-unbiased sample of 578 infrared (IR) quasars at z ≈ 1–3 and archival Atacama Large Millimetre/submillimetre Array submillimetre host galaxy sizes to investigate the ISM contribution to the quasar obscuration. We calculate star formation rates (SFR) and ISM column densities for the IR quasars and a control sample of submillimetre galaxies (SMGs) not hosting quasar activity and show that: (1) the quasar obscured fraction is constant up to |$\rm SFR\approx 300 \: {\rm M}_{\odot } \: yr^{-1}$|⁠ , and then increases towards higher SFR, suggesting that the ISM obscuration plays a significant role in starburst host galaxies, and (2) at |$\rm SFR\gtrsim 300 \: {\rm M}_{\odot } \: yr^{-1}$|⁠ , the SMGs and IR quasars have similarly compact submillimetre sizes (⁠|$R_{\rm e}\approx 0.5{\!-\!}3\,\mathrm{ kpc}$|⁠) and consequently, the ISM can heavily obscure the quasar, even reaching Compton-thick (⁠|$N_{\rm H}\gt 10^{24} \rm \: cm^{-2}$|⁠) levels in extreme cases. Based on our results, we infer that |${\approx} 10{\!-\!}30~{{ \rm per\ cent}}$| of the IR quasars with |$\rm SFR\gtrsim 300 \: {\rm M}_{\odot } \: yr^{-1}$| are obscured solely by the ISM. [ABSTRACT FROM AUTHOR]