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Genomic tools are now available for most livestock species and are used routinely for genomic selection (GS) in cattle. One of the most important developments resulting from the introduction of genomic testing for dairy cattle is the application of reasonably priced low-density single nucleotide polymorphism technology in the selection of females. In this context, combining genome testing and reproductive biotechnologies in young heifers enables new strategies to generate replacement and elite females in a given period of time. Moreover, multiple markers have been detected in biopsies of preimplantation stage embryos, thus paving the way to develop new strategies based on preimplantation diagnosis and the genetic screening of embryos. Based on recent advances in GS, the present review focuses on new possibilities inherent in reproductive technologies used for commercial purposes and in genetic schemes, possible side effects and beneficial impacts on reproductive efficiency. A particular focus is on the different steps allowing embryo genotyping, including embryo micromanipulation, DNA production and quality assessment. [ABSTRACT FROM AUTHOR]

Ideally, survey respondents read and understand survey instructions, questions, and response scales, and provide answers that carefully reflect their beliefs, attitudes, or knowledge. However, respondents may also arrive at their responses using cues or heuristics that facilitate the production of a response, but diminish the targeted information content. We use eye-tracking data as covariates in a Bayesian switching-mixture model to identify different response behaviors at the item-respondent level. The model distinguishes response behaviors that are predominantly influenced either positively or negatively by the previous response, and responses that reflect respondents' preexisting knowledge and experiences of interest. We find that controlling for multiple types of adaptive response behaviors allows for a more informative analysis of survey data and respondents., (© 2021. The Psychometric Society.)

The polycyclic aromatic hydrocarbon (PAH) benzo[a]pyrene (BP) is metabolized into a complex pattern of BP derivatives, among which the ultimate carcinogen (+)-anti-BP-7,8-diol-9,10-epoxide (BPDE) is formed to certain extents. Skin is frequently in contact with PAHs and data on the metabolic capacity of skin tissue toward these compounds are inconclusive. We compared BP metabolism in excised human skin, commercially available in vitro 3D skin models and primary 2D skin cell cultures, and analyzed the metabolically catalyzed occurrence of seven different BP follow-up products by means of liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). All models investigated were competent to metabolize BP, and the metabolic profiles generated by ex vivo human skin and skin models were remarkably similar. Furthermore, the genotoxicity of BP and its derivatives was monitored in these models via comet assays. In a full-thickness skin, equivalent BP-mediated genotoxic stress was generated via keratinocytes. Cultured primary keratinocytes revealed a level of genotoxicity comparable with that of direct exposure to 50-100 nM of BPDE. Our data demonstrate that the metabolic capacity of human skin ex vivo, as well as organotypic human 3D skin models toward BP, is sufficient to cause significant genotoxic stress and thus cutaneous bioactivation may potentially contribute to mutations that ultimately lead to skin cancer.

Background: Attention-deficit/hyperactivity disorder (ADHD) is a complex neurodevelopmental disorder characterized by deficits in attention, increased motor impulsivity, and hyperactivity. Preliminary work in mice and humans has suggested the X-linked gene STS (which encodes the enzyme steroid sulfatase) as a mediator of attentional functioning and as a candidate gene for ADHD., Methods: The effects of modulating the murine steroid sulfatase axis pharmacologically (through administration of the substrate dehydroepiandrosterone sulfate [DHEAS], 0-40 mg/kg, or acute inhibition of the enzyme by COUMATE, 10mg/kg) or genetically (through loss of the gene in 39,X(Y)*O mice) were assayed using the 5-choice serial reaction time task (5-CSRTT) a test of visuospatial attention and response control, and a locomotor activity paradigm., Results: DHEAS administration improved 5-CSRTT performance under attentionally demanding conditions, whereas steroid sulfatase inhibition impaired accuracy under the same conditions. Loss of Sts expression constitutively throughout development in 39,X(Y)*O mice resulted in deficits in 5-CSRTT performance at short stimulus durations and reduced anticipatory responding. Neither the pharmacologic nor the genetic manipulations affected basic locomotor activity., Conclusions: These data provide converging evidence indicating a role for steroid sulfatase in discrete aspects of attentional functioning and are suggestive of a role in motor impulsivity. The findings provide novel insights into the neurobiology of attention and strengthen the notion of STS as a candidate gene for the attentional component of ADHD.

We have constructed a computational platform suitable for examining emergence of shape homeostasis in simple three-dimensional cellular systems. An embryo phenotype results from a developmental process starting with a single cell and its genome. When coupled to an evolutionary search, this platform can evolve embryos with particular stable shapes and high capacity for self-repair, even though repair is not genetically encoded or part of the fitness criteria. With respect to the genome, embryo shape and self-repair are emergent properties that arise from complex interactions among cells and cellular components via signaling and gene regulatory networks, during development or during repair. This report analyzes these networks and the underlying mechanisms that control embryo growth, organization, stability, and robustness to injury.

Our approach to tissue modeling incorporates biologically derived primitives into a computational engine (CellSim) coupled with a genetic search algorithm. By expanding an evolved synthetic genome CellSim is capable of developing a virtual tissue with higher order properties. Using primitives based on cell signaling, gene networks, cell division, growth, and death, we have encoded a 64-cell cube-shaped tissue with emergent capacity to repair itself when up to 60% of its cells are destroyed. Other tissue shapes such as sheets of cells also repair themselves. Capacity for self-repair is an emergent property derived from, but not specified by, the rule sets used to generate these virtual tissues.

The objective of the present study was to investigate the correlation of soluble apoplastic peroxidase activity with lignification in needles of field-grown Norway spruce (Picea abies L.) trees. Apoplastic peroxidases (EC 1.11.1.7) were obtained by vacuum infiltration of needles. The lignin content of isolated cell walls was determined by the acetyl bromide method. Accumulation of lignin and seasonal variations of apoplastic peroxidase activities were studied in the first year of needle development. The major phase of lignification started after bud break and was terminated about 4 weeks later. This phase correlated with a transient increase in apoplastic guaiacol and coniferyl alcohol peroxidase activity. NADH oxidase activity, which is thought to sustain peroxidase activity by production of H2O2, peaked sharply after bud break and decreased during the lignification period. Histochemical localization of peroxidase with guaiacol indicated that high activities were present in lignifying cell walls. In mature needles, lignin was localized in walls of most needle tissues including mesophyll cells, and corresponded to 80 to 130 [mu]mol lignin monomers/g needle dry weight. Isoelectric focusing of apoplastic washing fluids and activity staining with guaiacol showed the presence of strongly alkaline peroxidases (isoelectric point [greater than or equal to] 9) in all developmental stages investigated. New isozymes with isoelectric points of 7.1 and 8.1 appeared during the major phase of lignification. These isozymes disappeared after lignification was terminated. A strong increase in peroxidase activity in autumn was associated with the appearance of acidic peroxidases (isoelectric point [less than or equal to] 3). These results suggest that soluble alkaline apoplastic peroxidases participate in lignin formation. Soluble acidic apoplastic peroxidases were apparently unrelated to developmentally regulated lignification in spruce needles.

Monoclonal antibodies that recognize individual polypeptides of the outer arm dyneins of Chlamydomonas flagella were obtained and used to study the structural relationships between the various polypeptides. Immunoblot analysis showed that the gamma heavy chain of 12S dynein and the alpha and beta heavy chains and Mr 69,000 intermediate chain of 18S dynein each contain immunoreactive sites not found in the other dynein chains or in any other axonemal protein. We also used these antibodies to investigate possible structural similarities between dynein polypeptides from Chlamydomonas and phylogenetically distant species. No crossreactivity was observed with antibodies against either the alpha, beta, or gamma heavy chains, demonstrating that each Chlamydomonas heavy chain has structural features distinct from those present in dyneins from the other species tested. However, one antibody against the Mr 69,000 polypeptide recognized an intermediate chain (Mr 76,000) of latent-activity dynein-1 from the sea urchin Tripneustes gratilla. This result provides further evidence that 18S dynein and latent-activity dynein-1 are related. In the course of the above studies, we modified existing procedures to achieve efficient transfer of high molecular weight proteins from NaDodSO4/polyacrylamide gels to nitrocellulose sheets, and to detect small quantities of protein on nitrocellulose. Our modified procedure for staining total protein on nitrocellulose is rapid, inexpensive, and as sensitive as silver-staining of polyacrylamide gels. These methods should be useful to investigators working with small amounts of protein or requiring resolution of closely migrating polypeptides after transfer to nitrocellulose.

Trifluoperazine (TFP), a drug that binds to Ca2+-calmodulin (CaM) complexes, altered swimming behavior not only in living paramecia, but also in reactivated, Triton-extracted "models" of the ciliate. By comparing the responses of living cells and models, we have ascertained that two sites of drug action exist in paramecium cilia. Swimming movements were recorded in darkfield stroboscopic flash photomicrographs; this permitted accurate quantitation of velocities and body-shape parameters. When living paramecia were incubated in a standard buffer containing 10 microM TFP, their speed of forward swimming fell over several minutes and their bodies shortened. Untreated paramecia backed up repeatedly and frequently upon transfer to a solution containing barium ions (the "barium dance"), but cells preincubated in TFP did not "dance." Instead they swam forward slowly for long periods of time without reversing and occasionally then exhibited abnormally prolonged reversals. W7 effects on swimming mimicked low doses of TFP, and the analog W5 did not visibly alter normal swimming patterns. These results suggest that TFP induces a decrease in the intracellular pCa of living paramecia, perhaps by reducing the efficiency of a calmodulin-activated calcium pump in the cell membrane. Paramecia extracted with Triton X-100 and reactivated to swim forward (7 greater than or equal to pCa greater than or equal to 6) were not affected by addition of up to 40 microM TFP to the reactivation medium. We conclude that the main drug effect in living cells is probably not at the axoneme. However, at low pCa, TFP directly affected the ciliary axoneme to shift its behavior to one characteristic of a higher pCa: TFP inhibited backward swimming in models reactivated at pCa less than 6; instead they swam forward or rocked in place. The mechanism of ciliary reversal in paramecium may therefore depend on an axonemal Ca2+-sensor, possibly bound CaM, which is affected by TFP only at low pCa, as has been postulated for other types of cilia.

We have developed conditions for the efficient electrotransfer from polyacrylamide gels to nitrocellulose sheets of a broad size range of proteins (Mr 8,000 to Mr greater than 400,000). The important features of this procedure include a two-step electrotransfer, beginning with elution of low-molecular-weight polypeptides at a low current density (approximately 1 mA/cm2) for 1 h, followed by prolonged electrotransfer (16-20 h) at high current density (approximately 3.5-7.5 mA/cm2) in conditions that favor the elution of high-molecular-weight proteins. The transfer buffer includes 0.01% sodium dodecyl sulfate to enhance protein elution, and 20% methanol to improve the retention of proteins on the nitrocellulose sheet. The nitrocellulose is air-dried after transfer is complete to eliminate loss of proteins during subsequent processing. This transfer procedure works well with proteins prepared from many different cell types, and is suitable for use with all polyacrylamide gel systems tested. With little or no modification, our method should also be applicable to transfer membranes other than nitrocellulose.

The behaviour of swimming Paramecium is markedly affected by hydrostatic pressure (50-200 atm, 1 atm = 101 325 Pa). To investigate whether pressure might alter behaviour by acting directly on specific ion channels that mediate the behavioural responses, we examined the effects of K+, Na+ and Ba2+ ions on swimming speed and the reversal response during pressurization and decompression. If pressure acted on the channels that transport these ions, then the pressure-induced responses of swimming Paramecium should be exaggerated or diminished, according to which ions were present in the experimental buffer. Pressurization to 100 atm in standard buffer inhibited the brief reversal of swimming direction that occurred at atmospheric pressure when a paramecium encountered the wall of the pressure chamber. To determine whether pressure impaired mechanoreceptor function or directly blocked the Ca2+-channels that control ciliary reversal, we added Ba2+ or Na+ to standard buffer to induce multiple spontaneous reversals. Pressurization blocked these reversals, suggesting that channel opening is directly inhibited by pressure. Decompression in standard buffer elicited momentary ciliary reversal and backward swimming. Buffers with a high ratio of K+ to Ca2+ suppressed this response, and the decompression-induced reversal was exaggerated in the presence of Ba2+ or Na+, consistent with the effects that these ions are known to have on Paramecium's reversal response. These data imply that, upon decompression, the Ca2+-channels that mediate ciliary reversal open transiently. In addition to blocking the reversal response, pressurization slowed forward swimming. By examining the response to pressurization of Paramecium immobilized by Ni2+, we found that hydrostatic pressure apparently slows swimming by reorientating the direction of ciliary beat.