Later Phases Of Development Research Articles

Recapitulation of Neural Crest Specification and EMT via Induction from Neural Plate Border-like Cells

SummaryNeural crest cells (NCCs) contribute to several tissues during embryonic development. NCC formation depends on activation of tightly regulated molecular programs at the neural plate border (NPB) region, which initiate NCC specification and epithelial-to-mesenchymal transition (EMT). Although several approaches to investigate NCCs have been devised, these early events of NCC formation remain largely unknown in humans, and currently available cellular models have not investigated EMT. Here, we report that the E6 neural induction protocol converts human induced pluripotent stem cells into NPB-like cells (NBCs), from which NCCs can be efficiently derived. NBC-to-NCC induction recapitulates gene expression dynamics associated with NCC specification and EMT, including downregulation of NPB factors and upregulation of NCC specifiers, coupled with other EMT-associated cell-state changes, such as cadherin modulation and activation of TWIST1 and other EMT inducers. This strategy will be useful in future basic or translational research focusing on these early steps of NCC formation.

Effects of interleukin-6 and tumor necrosis factor-α on the expression of angiogenic and collagenolytic factors in premature and mature adipocytes

ObjectiveThe remodeling of the vascular network and collagen in the extracellular matrix is closely associated with the expansion and dysfunction of adipose tissue. In the present study, we investigated the effects of interleukin (IL)-6 and tumor necrosis factor (TNF)-α on the expression of angiogenic factors, collagen, and collagenase and its endogenous inhibitor in premature and mature adipocytes. MethodsPremature and mature adipocytes were differentiated from 3T3-L1 cells and stimulated with IL-6 or TNF-α to mimic the early and late phases of obesity development. The levels of expression of angiogenic factors, including vascular endothelial cell growth factor a (Vegfa), hepatocyte growth factor (Hgf), angiopoietin (Angpt)1, and Angpt2, as well as type I collagen, matrix metallopeptidase (Mmp) 13, and tissue inhibitor of Mmp (Timp) 1, were determined using real-time reverse transcription polymerase chain reaction or enzyme-linked immunosorbent assay. Human umbilical vein endothelial cells were grown with the culture supernatant of adipocytes stimulated with/without IL-6 or TNF-α, and the formation of tube structures was evaluated. ResultsIL-6 and TNF-α induced the expression of Vegfa, Hgf, and Angpt2 and decreased the expression of Angpt1 in premature adipocytes, whereas, they decreased the expression of Vegfa and Hgf in mature adipocytes. The culture supernatant of IL-6- or TNF-α-stimulated premature adipocytes induced the formation of tube structures. IL-6 and TNF-α had no effects on type I collagen expression in both premature and mature adipocytes but suppressed the expression of Mmp13 and Timp1 in mature and premature adipocytes, respectively. ConclusionThe effects of IL-6 and TNF-α on the expression of angiogenic and collagenolytic factors differed between premature and mature adipocytes. This finding suggests that these inflammatory cytokines induce expansion and dysfunction of adipose tissue via angiogenesis and collagen turnover in premature and mature adipocytes.

An easy long-acting BMP7 release system based on biopolymer nanoparticles for inducing osteogenic differentiation of adipose mesenchymal stem cells.

In contrast to the early acting bone morphogenetic protein 2, bone morphogenetic protein 7 (BMP7) plays a decisive role mainly in the late stages of bone formation. To overcome deactivation and degradation of expensive BMP7, we designed a novel long-acting BMP7 release system based on poly(3-hydroxybutyrate-co-4-hydroxybutyrate) (P34HB) nanoparticles to enable the induction of osteogenic differentiation in human adipose mesenchymal stem cells (ADSCs). In order to improve the encapsulation efficiency of BMP7 and avoid damage by organic solvents, BMP7 was modified and protected using the biosurfactant soybean lecithin. In an in vitro test, BMP7-soybean lecithin-P34HB nanoparticles (BMP7-SPNPs) showed a short initial burst of BMP7 release during the first 24h, followed by a steady increase to a cumulative 80% release in 20days. Compared with the rapid release of control P34HB nanoparticles without soybean phospholipids loaded with BMP7 without soybean lecithin, BMP7-SPNPs significantly reduced the initial burst of BMP7 release and stabilized the content of BMP7 to allow long-term osteogenic differentiation during the late phase of bone development. Human ADSCs treated with BMP7-SPNPs showed higher alkaline phosphatase activity and higher expression levels of genetic markers of osteogenic differentiation compared with the control group. Thus, the results indicate that BMP7-SPNPs can be used as a rapid and long-acting BMP7 delivery system for osteogenic differentiation.

The legacy of management approaches and abandonment on old-growth attributes in hardwood floodplain forests in the Pannonian Ecoregion

In the absence of primeval floodplain forests, near-natural remnants are key references for close-to-nature forestry and nature conservation. Old-growth forest characteristics (OGCs) were quantified in 16 managed (rotation or selection) and abandoned semi-natural floodplain forests by taking snapshot inventories of structural features. Principal component analysis revealed a short gradient from closed-canopy managed stands dominated by large trees (mainly oaks) and hornbeams, with less deadwood, to abandoned older stands with more gaps, very large trees and different kinds of deadwood. All the studied forests are mainly in the early and late biostatic phases of forest development, with innovation and degradation phases almost completely absent. Results showed that selection forests had significantly more very large trees compared to rotation stands. The difference in structure between abandoned selection stands and abandoned rotation stands was more pronounced than in their counterparts that are still managed. Based on diameter distribution and the amount of very large trees and deadwood, abandoned selection stands were closest to the structure of natural forests. Selection forests may recover certain OGCs more rapidly than rotation forests. We concluded that after 100–150 years of homogenising rotation forestry, OGCs are slow to develop in just 25 years of non-intervention. Based on our results, we recommend: (i) an increase in the rotation cycle with selective logging in order to restore and maintain OGCs in managed rotation forests and (ii) total abandonment in certain well-preserved, near-natural forests with natural flooding regimes. However, the failure of oak to regenerate naturally remains a challenge.

A panel of criteria for comprehensive assessment of severity of ultraviolet B radiation-induced non-melanoma skin cancers in SKH-1 mice

The study of causes and cures for ultraviolet B radiation (UVB)-induced non-melanoma skin cancers (NMSC) has been greatly facilitated by use of the albino SKH-1 hairless mice. These mice develop multiple tumors of different sizes and the severity of cancer is often measured by one or more of the four criteria, namely the prevalence, multiplicity, area and volume of tumors. However, there are inherent limitations of each criterion: the prevalence and number do not account for size differences among tumors, area measurement ignores the tumor height, and volume measurement overcompensates for the height at the cost of planar dimensions. Here, using our dataset from an ongoing NMSC study, we discuss the limitations of these four criteria, and suggest refinements in measuring prevalence. We recommend the use of three more criteria, namely the Knud Thomsen tridimensional surface that apportions optimal weightage to three tumor dimensions, weekly occurrence of new tumors and tumor growth-rate to reveal initiation and growth of tumors in early and late phase of NMSC development, respectively. Together, use of this comprehensive panel of seven criteria can provide an accurate assessment of severity of NMSC and lead to a testable hypothesis whether the experimental manipulation of mice has affected the early initiation or growth phase of NMSC tumors.

Heat shock factor 5 is essential for spermatogenesis in mice: Detected by a new monoclonal antibody.

Objective(s):Here, we examined the function of our produced monoclonal antibody (mAb10C3) to recognize one of the most important members of the HEAT shock factor family, Hsf5, in embryonic development and in spermatogenic cells of adult mouse testis.Materials and Methods:The targeting effects of mAb10C3 were investigated by immunohistochemistry analysis in the different phases of the embryo and in the adult testis tissue sections.Results:The results of immunohistochemistry staining on the mouse embryos by the supernatant of hybridoma clone that produced mAb10C3, in the early and late phases (E7.5 and E14.5) of embryonic development, indicated that mAb10C3 could only detect Hsf5 in E7.5 and it did not have any targeting activity in the late phase of development. Therefore, we showed that the hsf5 gene has expressed in early mouse embryonic development. On the other hand, mAb10C3 could detect Hsf5 in spermatogonia and spermatocytes of adult testis in comparison with a known anti-Hsf5 antibody (ab98939) and an anti-PCNA antibody as a marker of spermatogonia cells.Conclusion:Taken together, these data indicated that generated anti-testis mAb10C3 was generated against anti-testis proteins, specifically to target Hsf5, and can be useful as a scientific tool to investigate the critical genes in the development and spermatogenesis.

Pou3f4-expressing otic mesenchyme cells promote spiral ganglion neuron survival in the postnatal mouse cochlea.

During inner ear development, primary auditory neurons named spiral ganglion neurons (SGNs) are surrounded by otic mesenchyme cells, which express the transcription factor Pou3f4. Mutations in Pou3f4 are associated with DFNX2, the most common form of X-linked deafness and typically include developmental malformations of the middle ear and inner ear. It is known that interactions between Pou3f4-expressing mesenchyme cells and SGNs are important for proper axon bundling during development. However, Pou3f4 continues to be expressed through later phases of development, and potential interactions between Pou3f4 and SGNs during this period had not been explored. To address this, we documented Pou3f4 protein expression in the early postnatal mouse cochlea and compared SGNs in Pou3f4 knockout mice and littermate controls. In Pou3f4y/- mice, SGN density begins to decline by the end of the first postnatal week, with approximately 25% of SGNs ultimately lost. This period of SGN loss in Pou3f4y/- cochleae coincides with significant elevations in SGN apoptosis. Interestingly, this period also coincides with the presence of a transient population of Pou3f4-expressing cells around and within the spiral ganglion. To determine if Pou3f4 is normally required for SGN peripheral axon extension into the sensory domain, we used a genetic sparse labeling approach to track SGNs and found no differences compared with controls. We also found that Pou3f4 loss did not lead to changes in the proportions of Type I SGN subtypes. Overall, these data suggest that otic mesenchyme cells may play a role in maintaining SGN populations during the early postnatal period.

Parameter estimation and evaluation of Ross-Li and RPV models for wheat phenophases using hemispherical directional reflectance measurements

ABSTRACT Hemispherical directional reflectance factor (HDRF) measurements of wheat crop acquired with field goniometer system are used to estimate parameters and evaluate anisotropic reflectance models. The HDRFs were sampled at 54 view angles (6 azimuthal planes × 9 view zenith angles) to cover the hemispheric span up to 60° zenith. The observations were made at 7–8 days’ interval during different phenophases of the crop. Biophysical parameters were also measured parallel to HDRF. Kernel driven Ross-Li variants and RPV (Rehman Pinty Verstrate) models parameters are generated by fitting HDRF data. RossThin-H/LiSparse-R model was fitted best for all visible-near infrared bands in early phenophases while the RossThick-H/LiSparse-R model was best fitted for heading and flowering phases in most cases. In late grain development phases (Late milking and Dough) of the wheat, HDRFs of visible bands were best fitted with RossThin-H/LiDense-R model in most instances. The HDRF simulation of wheat by RPV models is identical to Ross-Li models. Both models are adequate to account reflectance anisotropy of wheat canopy. Phenophase specific model parameters can be used to reconstruct reflectance anisotropy of wheat for different applications.

Application and Impact of Human Dose Projection from Discovery to Early Drug Development.

The application and impact of human dose projection (HDP) has been well recognized in the late drug development phase, with increasing appreciation earlier during discovery and early development. This commentary describes the perspective of pharmaceutical scientists on the evolving application and impact of HDP at various phases from discovery to early development, including lead generation, lead optimization, lead up to candidate nomination, and early drug development. The underlying fundamental concepts and key input parameters for HDP are briefly discussed. A broad overview of phase-specific tools and approaches commonly utilized for human dose projection in the pharmaceutical industry is provided. A discussion of phase-appropriate implementation strategies, associated limitations/assumptions and continuous refinement for HDP from discovery to early development is presented. The authors describe the phase-specific applications of human dose projection to facilitate key assessments and relative impact on decision points.

Dynamic Mergers Effects on R&D Investments and Drug Development Across Research Phases in the Pharmaceutical Industry

Pharmaceutical firms spend increasing amounts in mergers and acquisitions (M&As), which raises the question of whether sufficient investment is left after mergers to further develop firms’ internal drug development capability. We evaluate the effects of M&As on firms’ post-merger R&D investments and drug development capabilities across drug development phases. This study builds on a novel database that enables us to evaluate the post-merger effect at the research project level and across development phases. A further novel feature of the study is allowing measurement errors to enter firms’ R&D investments. Our study adopts a structural equation modeling approach, which is appropriate for evaluating a system of equations through which we examine the direct and indirect merger effects on R&D capabilities across development phases. We find that M&As have a strong effect on firms’ drug development at the late development phases through economies of scope. At the early development phases, M&As serve to replenish firms’ drug pipelines. The study shows that M&As have a direct and negative effect on firms’ R&D investments. However, the overall effect on R&D investments accounting for enhanced post-merger R&D capabilities and product approvals turns out to be positive. M&As can be an effective instrument for firms to acquire drug development knowledge and technology in late stages of the development process (Phases 3 clinical testing and regulatory filing). Our study provide empirical evidence that investments in M&As in late stage of drug development help firms’ growth and increase firms revenue.

УПРАВЛІННЯ ІННОВАЦІЙНИМ РОЗВИТКОМ ПІДПРИЄМСТВА ІЗ ЗАСТОСУВАННЯМ ТЕХНОПОЛІСНОЇ КОНЦЕПЦІЇ

The article provides a research survey in the area of innovation development and innovation process management. It offerstechno-policy model of information maintenance of innovation development. А соnсеpt is given related to industrial andadministrative-territorial information flows synthesis to form а common base of knowledge in the field of management andinnovation development. The growing use of new knowledge in a developed system of innovation processes has become thebasis for sustainable socio-economic development of countries that are entering the late industrial and post-industrial phases ofdevelopment. The analysis of current trends and the role of this new sector of the world economy, conducted by the authors,gives grounds for the following conclusions: – the results of scientific and technical (innovative) activities are one of the mainresources that determines the rate of economic growth of developed countries; – the role of the new sector in the distribution ofspheres of influence on the international markets of tangible assets (production volume and profit from their sale) is growing; –the role of the market of innovative products in the redistribution of price policy factors between industrialized countries andcountries exporting raw materials (through a system of world prices for raw materials and prices for products made usingknowledge-intensive and high technology). It is fundamentally important that the effectiveness of development in modern societyis achieved not only through the development of science and industry, but also by ensuring the entire innovation cycle, from theprocess of acquiring new knowledge, transferring it to productive sectors of the economy and ending with their use andimplementation. Since the use of new knowledge is usually carried out in order to obtain economically significant competitiveadvantages in the final product market, all stages of the innovation cycle are under the informational, organizational and financialinfluence of market factors.

Dynamics of the microbial community during growth of the house dust mite Dermatophagoides farinae in culture.

The variation in house dust mite microbial communities is important because various microorganisms modulate the production of allergens by their mite hosts and/or contaminate immunotherapeutic extracts. Temporal changes in mite microbiomes and the mite culture environment occurring at different stages of mite culture development are particularly understudied in this system. Here, we analyzed the dynamics of microbial communities during the culture growth of Dermatophagoides farinae. Changes in microbiomes were related to three key variables: the mite population density, microbial microcosm respiration and concentration of guanine (the mite nitrogenous waste metabolite). Mite populations exhibited the following phases: exponential growth, plateau and exponential decline. The intracellular bacterium Cardinium and the yeast Saccharomyces cerevisiae prevailed in the internal mite microbiomes, and the bacterium Lactobacillus fermentum was prevalent in the mite diet. The reduction in the mite population size during the late phases of culture development was related to the changes in their microbial profiles: the intracellular bacterium Cardinium was replaced by Staphylococcus, Oceanobacillus and Virgibacillus, and S. cerevisiae was replaced by the antagonistic fungi Aspergillus penicillioides and Candida. Increases in the guanine content were positively correlated with increases in the Staphylococcus and A. penicillioides profiles in the culture environment. Our results show that the mite microbiome exhibits strong, dynamic alterations in its profiles across different mite culture growth stages.

The velvet repressed vidA gene plays a key role in governing development in Aspergillus nidulans.

Fungal development is regulated by a variety of transcription factors in Aspergillus nidulans. Previous studies demonstrated that the NF-κB type velvet transcription factors regulate certain target genes that govern fungal differentiation and cellular metabolism. In this study, we characterize one of the VosA/VelB-inhibited developmental genes called vidA, which is predicted to encode a 581-amino acid protein with a C2H2 zinc finger domain at the C-terminus. Levels of vidA mRNA are high during the early and middle phases of asexual development and decrease during the late phase of asexual development and asexual spore (conidium) formation. Deletion of either vosA or velB results in increased vidA mRNA accumulation in conidia, suggesting that vidA transcript accumulation in conidia is repressed by VosA and VelB. Phenotypic analysis demonstrated that deletion of vidA causes decreased colony growth, reduced production of asexual spores, and abnormal formation of sexual fruiting bodies. In addition, the vidA deletion mutant conidia contain more trehalose and β-glucan than wild type. Overall, these results suggest that VidA is a putative transcription factor that plays a key role in governing proper fungal growth, asexual and sexual development, and conidia formation in A. nidulans.

Arabidopsis IPGA1 is a microtubule-associated protein essential for cell expansion during petal morphogenesis.

Unlike animal cells, plant cells do not possess centrosomes that serve as microtubule organizing centers; how microtubule arrays are organized throughout plant morphogenesis remains poorly understood. We report here that Arabidopsis INCREASED PETAL GROWTH ANISOTROPY 1 (IPGA1), a previously uncharacterized microtubule-associated protein, regulates petal growth and shape by affecting cortical microtubule organization. Through a genetic screen, we showed that IPGA1 loss-of-function mutants displayed a phenotype of longer and narrower petals, as well as increased anisotropic cell expansion of the petal epidermis in the late phases of flower development. Map-based cloning studies revealed that IPGA1 encodes a previously uncharacterized protein that colocalizes with and directly binds to microtubules. IPGA1 plays a negative role in the organization of cortical microtubules into parallel arrays oriented perpendicular to the axis of cell elongation, with the ipga1-1 mutant displaying increased microtubule ordering in petal abaxial epidermal cells. The IPGA1 family is conserved among land plants and its homologs may have evolved to regulate microtubule organization. Taken together, our findings identify IPGA1 as a novel microtubule-associated protein and provide significant insights into IPGA1-mediated microtubule organization and petal growth anisotropy.

Acute Myeloid Leukemia Mutations: Therapeutic Implications.

Acute Myeloid Leukemia (AML) is an extremely heterogeneous group of hematological neoplasms, for which allogeneic stem cell transplantation (HSCT) still represents the only potentially curative option in the majority of cases. However, elderly age and clinically severe comorbidities may often exclude a wide amount of patients from this therapeutic approach, underlying the urgent need for alternative strategies. Thanks to the introduction of advanced high-throughput techniques, light is being shed on the pathogenesis of AML, identifying molecular recurrent mutations as responsible for the onset, as well as progression, of disease. As a consequence, and in parallel, many new compounds, including targeted therapies (FMS-like tyrosine kinase 3 (FLT3) and Isocitrate dehydrogenase 1-2 (IDH1-2) inhibitors), have found a wide room of application in this setting, and are now available in daily practice, or in late phases of clinical development. Moreover, several further innovative molecules are currently under investigation, and promising results for many of them have already been reported. In this review, we will present an update on the most relevant molecular alterations of AML, focusing on the most frequent genomic mutations of the disease, for which compounds have been approved or are still currently under investigation.

Morphological, molecular evolution an in vitro digestibility of filamentous granules of banana starch during fruit development.

The development of starch granules of a banana cultivar (morado variety - Musa AAA subgroup Red dacca) from filamentous shape to semi-spherical and finally to oval shape, was studied. The purity of the extracted starch changed from 83.5% (6 weeks) to 95.4% (16 weeks). Impurities were ascribed to cellulosic and latex fractions responsible for the integrity of the pristine fruit. The amylose content was stabilized at about 29.6% after the 12th week. The thermal analysis showed that the gelatinization enthalpy increased from 5.0 to 11.2 J/g from the 6th to the 12th week, indicating an increased degree of internal molecular organization. The analysis of chain-length distribution and gel permeation chromatography, showing that the content of long chains (B1, B2, and B3+) increased with the development of the starch granule. Also, XRD analysis indicated that C- type X-ray diffraction pattern from early to later phases of development, although the relative crystallinity content increased from 19.3 to 23.5% after 16 weeks of development. FTIR revealed the formation of more ordered structures with the development time. In vitro digestion tests showed that the resistant starch fraction increased from 37.5% for week 6 to 55.5% for week 16.

Digital technologies of investment analysis of projects for the development of oil fields of unallocated subsoil reserve fund

The task of objective economic estimation of unallocated subsoil reserve fund (USRF) objects is especially important for regions characterized by late phase of development and aggravation of mining-and-geological modes of occurrence of deposits to be implemented. This article presents an information-analytic technology for evaluation of investment prospects of USRF oil fields. This technology is a methodological base for the technical-and-economic estimation with regard to specific industrial aspects, such as: probability estimates of confirmed reserves; determination of primary technological oil field development parameters, etc. Making techno economic estimations is possible in case of using specialized software which allows to process big data, such as EVA (economic estimation of oil fields), EVA-Risk, etc. The suggested info-analytic technology has been practically tested and parameters of investment prospects have been determined for USRF oil fields in the Komi Republic which is the region having highly developed oil and gas sector and unstable prognosis regarding oil extraction for the nearest decade.

Fine characterization of large composite channel sandbody architecture and its control on remaining oil distribution: A case study of alkaline-surfactant-polymer (ASP) flooding test area in Xingshugang oilfield, China

Lateral accretion deposit, the most critical element in channel sandbody architecture, significantly affect the distribution of remaining oil in the middle and late development phases of oilfields. However, the lateral accretion deposits in a composite channel sandbody are extremely thin and small, complicating the identification through log data. In this regard, the fine characterization of sandbody architecture is an effective alternative, which can further guide the evaluation of the remaining oil distribution in composite channels. In this paper, illustrated by an example of the alkaline-surfactant-polymer (ASP) flooding test area of Xing 12 block in Daqing oil field, China, a fine characterization method of the sandbody architecture of large composite channel was proposed. Integrated with the production data, the influence of the lateral accretion deposits of the sandbody on the direction and uniformity of waterflooding was analyzed, and the distribution pattern of the remaining oil controlled by sandbody architecture was discussed. The results show that, although the sandbodies in the large composite channel seem to have characteristics of connectivity and efficiency as a whole, the sandbody is only an apparent connecting body in reality. Due to the complicated contact relationship of the various architectural elements at all levels and stages, and the existence of low permeability barriers, for instance, thin lateral accretion beds and abandoned channel, the sandbodies of the large composite channel are not completely connected. The fifth and third level of architectural interfaces in the sandbody architecture of large composite channels control the frame of non-permeable lateral accretion deposits. Blocked by these lateral accretion deposits, the thickness of watered out reservoirs decreases along the lateral accretion direction, in which the bottom of the point bar is watered out severely. Hence the remaining oil is mainly distributed in the middle and upper parts of the composite channels, the inter-channel sands where the production and injection wells are imperfect, as well as the edge of channels. Under the control of the sandbody architecture of large composite channels, three types of remaining oil were determined: remaining oil blocked by lateral accretion units, remaining oil blocked by abandoned channel and remaining oil located at the edge of the channel with poor petrophysical properties.

The Landscape of Structural Variant Signatures in Multiple Myeloma Identifies Distinct Disease Subgroups with Implications for Pathogenesis

The Landscape of Structural Variant Signatures in Multiple Myeloma Identifies Distinct Disease Subgroups with Implications for Pathogenesis

Characterization of the velvet regulators in Aspergillus flavus.

Fungal development and secondary metabolism are closely associated via the activities of the fungal NK-kB-type velvet regulators that are highly conserved in filamentous fungi. Here, we investigated the roles of the velvet genes in the aflatoxigenic fungus Aspergillus flavus. Distinct from other Aspergillus species, the A. flavus genome contains five velvet genes, veA, velB, velC, velD, and vosA. The deletion of velD blocks the production of aflatoxin B1, but does not affect the formation of sclerotia. Expression analyses revealed that vosA and velB mRNAs accumulated at high levels during the late phase of asexual development and in conidia. The absence of vosA or velB decreased the content of conidial trehalose and the tolerance of conidia to the thermal and UV stresses. In addition, double mutant analyses demonstrated that VosA and VelB play an inter-dependent role in trehalose biosynthesis and conidial stress tolerance. Together with the findings of previous studies, the results of the present study suggest that the velvet regulators play the conserved and vital role in sporogenesis, conidial trehalose biogenesis, stress tolerance, and aflatoxin biosynthesis in A. flavus.