Stages Of Cycle Research Articles

Concurrent D-loop cleavage by Mus81 and Yen1 yields half-crossover precursors.

Homologous recombination involves the formation of branched DNA molecules that may interfere with chromosome segregation. To resolve these persistent joint molecules, cells rely on the activation of structure-selective endonucleases (SSEs) during the late stages of the cell cycle. However, the premature activation of SSEs compromises genome integrity, due to untimely processing of replication and/or recombination intermediates. Here, we used a biochemical approach to show that the budding yeast SSEs Mus81 and Yen1 possess the ability to cleave the central recombination intermediate known as the displacement loop or D-loop. Moreover, we demonstrate that, consistently with previous genetic data, the simultaneous action of Mus81 and Yen1, followed by ligation, is sufficient to recreate the formation of a half-crossover precursor in vitro. Our results provide not only mechanistic explanation for the formation of a half-crossover, but also highlight the critical importance for precise regulation of these SSEs to prevent chromosomal rearrangements.

Effects of an indole derivative on cell proliferation, transfection, and alternative splicing in production of lentiviral vectors by transient co-transfection.

Lentiviral vectors derived from human immunodeficiency virus type I are widely used to deliver functional gene copies to mammalian cells for research and gene therapies. Post-transcriptional splicing of lentiviral vector transgene in transduced host and transfected producer cells presents barriers to widespread application of lentiviral vector-based therapies. The present study examined effects of indole derivative compound IDC16 on splicing of lentiviral vector transcripts in producer cells and corresponding yield of infectious lentiviral vectors. Indole IDC16 was shown previously to modify alternative splicing in human immunodeficiency virus type I. Human embryonic kidney 293T cells were transiently transfected by 3rd generation backbone and packaging plasmids using polyethyleneimine. Reverse transcription-quantitative polymerase chain reaction of the fraction of unspliced genomes in human embryonic kidney 293T cells increased up to 31% upon the indole's treatment at 2.5 uM. Corresponding yield of infectious lentiviral vectors decreased up to 4.5-fold in a cell transduction assay. Adjusting timing and duration of IDC16 treatment indicated that the indole's disruption of early stages of transfection and cell cycle had a greater effect on exponential time course of lentiviral vector production than its reduction of post-transcriptional splicing. Decrease in transfected human embryonic kidney 293T proliferation by IDC16 became significant at 10 uM. These findings indicated contributions by early-stage transfection, cell proliferation, and post-transcriptional splicing in transient transfection of human embryonic kidney 293T cells for lentiviral vector production.

Comparative Life Cycle Assessment of Electric and Internal Combustion Engine Vehicles

This article is devoted to the ecological comparison of electric and internal combustion engine vehicles throughout their entire life cycle, from mining to recycling. A scientifically based approach to a comprehensive environmental assessment of the impact of vehicles on the environment has been developed. To analyze the impact on the environmental situation, aspects such as the consumption of natural resources, waste generation, electricity consumption, emission of harmful substances into the atmosphere, water consumption, and greenhouse gas emissions are taken into consideration. As a result of comparing the environmental impacts of vehicles, it was found that natural resources consumption and production of industrial waste from electric vehicles (EV) is 6 times higher than from internal combustion engine vehicles (ICEV). Harmful substance emissions and greenhouse gas emissions from EV production are 1.65 and 1.5 times higher, respectively. The EV total electricity consumption is 1.4 times higher than that of ICEVs. At the same time, it was revealed that during operation, EVs have higher energy consumption and emit more harmful substances into the atmosphere, but EVs produce less greenhouse gas emissions. It means that at different life cycle stages, EVs have a much higher negative impact on the environment compared to gasoline engine vehicles.

Pharmacologic blockade of nicotinic receptors in the suprachiasmatic nucleus increases ovarian atresia and inhibits follicular growth.

Reproduction in all mammalian species depends on the growth and maturation of ovarian follicles, that is, folliculogenesis. Follicular development can culminate with the rupture of mature follicles and the consequent expulsion of their oocytes (ovulation) or in atresia, characterized by the arrest of development and eventual degeneration. These processes are regulated by different neuroendocrine signals arising at different hypothalamic nuclei, including the suprachiasmatic nucleus (SCN). In the later, the activation of muscarinic receptors (mAChRs) and nicotinic receptors (nAChRs) by acetylcholine is essential for the regulation of the pre-ovulatory signals that stimulate the rupture of mature follicles. To evaluate the participation of the nAChRs in the SCN throughout the oestrous cycle in the regulation of the hypothalamic-pituitary-ovarian axis. For this purpose, 90-day-old adult female rats in metoestrus, dioestrus, proestrus or oestrus were microinjected into the left- or right-SCN with 0.3 μL of saline solution as vehicle or with 0.225 μg of mecamylamine (Mec), a non-selective antagonist of the nicotinic receptors, diluted in 0.3 μL of vehicle. The animals were sacrificed when they presented vaginal cornification, indicative of oestrus stage, and the effects of the unilateral pharmacological blockade of the nAChRs in the SCN on follicular development, ovulation and secretion of oestradiol and follicle-stimulating hormone (FSH) were evaluated. The microinjection of Mec decreased the serum levels of FSH, which resulted in a lower number of growing and healthy follicles and an increase in atresia. The higher percentage of atresia in pre-ovulatory follicles was related to a decrease in the number of ova shed and abnormalities in oestradiol secretion. We also detected asymmetric responses between the left and right treatments that depended on the stage of the oestrous cycle. The present results allow us to suggest that during all the stages of the oestrous cycle, cholinergic signals that act on the nAChRs in the SCN are pivotal to modulate the secretion of gonadotropins and hence the physiology of the ovaries. Further research is needed to determine if such signals are generated by the cholinergic neurons in the SCN or by cholinergic afferents to the SCN.

Quantum Fuzzy Decision-Making for Analyzing the Service Progress Life Cycle of Renewable Energy Innovation Investments

Innovative and effective solutions can be produced by using advanced technologies to increase renewable energy innovation investments. However, many actions taken to increase these investments may cause the costs of enterprises to reach an unmanageable level. As a result, a priority analysis is needed to identify the most critical issues, but in literature, there are limited studies in the literature regarding this issue. Accordingly, the purpose of this study is to evaluate the service progress life cycle of renewable energy innovation investments. A novel model is presented in this process while integrating different approaches. First, the service progress life cycle of renewable innovation investments is weighted. In this scope, two-generation technology S-curve and facial action coding system-based Quantum Spherical fuzzy M-SWARA are considered. Secondly, the renewable innovation investment alternatives are ranked. In this framework, the integer patterns and facial action coding system-based Quantum Spherical fuzzy TOPSIS are taken into consideration. Additionally, these investment alternatives are also ranked by using VIKOR technique to test the validity of the proposed model results. Moreover, a sensitivity analysis is conducted by considering 8 different cases so that coherency of the results can be tested. The main contribution of this manuscript is the consideration of the FACS system to increase both the originality and effectiveness of the proposed model and the development of new technique named M-SWARA. Additionally, conducting a priority evaluation helps to identify the efficient investment strategies to increase renewable energy innovation. It is defined that the same findings can be achieved by both TOPSIS and VIKOR techniques. Furthermore, sensitivity analysis also indicates the same rankings as well. It is understood that the proposed model provides coherent and reliable findings. The findings denote that maturity is the most critical stage of the service progress life cycle of the renewable energy investments. Moreover, advanced technologies are found as the best alternative of the renewable innovation investments for the service progress life cycle. Hence, it is recommended that R&D activities should be prioritized for the provision of advanced technologies in the renewable energy sector. For this purpose, research centers, universities, private sector companies and government institutions can work for the development of innovative and advanced technologies.

An age-related nomogram for antral follicle counts as a reference guide for reproductive decision making: an observational study of 7573 women

Research questionTo investigate the relationship between antral follicle count (AFC) and chronological age and establish expected values for AFC. DesignThis is a retrospective cohort study at a specialist gynaecological ultrasound centre in London. Women presenting to a gynaecology diagnostic unit for investigation of symptoms or routine check-up, and undergoing transvaginal ultrasound examinations, between the 1st of January 2017 and 22nd of September 2022. ResultsWe analysed 8821 data entries from 7573 women. We estimated the relationship between AFC and age to develop an AFC nomogram independent of the stage in the menstrual cycle. The AFC corresponding to the 10th, 25th, 50th, 75th and 90th centiles for each patient and age group have been calculated. Both nomogram and condition specific populations displayed a gaussian relationship between AFC and age. For the nomogram population, a peak median AFC of 30, was observed between the age groups of 21 -23. The AFC distribution of the previous cystectomy group was found to be statistically different in comparison to the nomogram population(P<0.0001). However, the same did not hold true for those on hormonal contraception (P=0.43). ConclusionsWe provide an AFC nomogram reporting median and interquartile values for AFC by chronological age across the reproductive years. This is a useful tool for providing counselling for those undergoing ovarian reserve assessments taken any time in the menstrual cycle including for those on hormonal contraceptives or previous cystectomy.

Challenges and Potential of Artificial Intelligence in Neuroradiology.

Artificial intelligence (AI) has emerged as atransformative force in medical research and is garnering increased attention in the public consciousness. This represents acritical time period in which medical researchers, healthcare providers, insurers, regulatory agencies, and patients are all developing and shaping their beliefs and policies regarding the use of AI in the healthcare sector. The successful deployment of AI will require support from all these groups. This commentary proposes that widespread support for medical AI must be driven by clear and transparent scientific reporting, beginning at the earliest stages of scientific research. Areview of relevant guidelines and literature describing how scientific reporting plays acentral role at key stages in the life cycle of an AI software product was conducted. To contextualize this principle within aspecific medical domain, we discuss the current state of predictive tissue outcome modeling in acute ischemic stroke and the unique challenges presented therein. Translating AI methods from the research to the clinical domain is complicated by challenges related to model design and validation studies, medical product regulations, and healthcare providers' reservations regarding AI's efficacy and affordability. However, each of these limitations is also an opportunity for high-impact research that will help to accelerate the clinical adoption of state-of-the-art medical AI. In all cases, establishing and adhering to appropriate reporting standards is an important responsibility that is shared by all of the parties involved in the life cycle of aprospective AI software product.

Unfolding the role of cell state on stress granule heterogeneity and function in pancreatic cancer.

e16357 Background: A major impediment to cancer therapy is the reality that cancer is a heterogeneous disease. A single tumor can consist of many genotypic or phenotypic profiles, which are marked by degrees of sensitivity to different therapeutics. Identifying and targeting origins of cancer cell heterogeneity is critical for predicting patient outcome and enhancing treatment efficacy. We observe a novel source of cancer cell heterogeneity that involves one of the largest stress-sensing platforms described to date: the stress granule (SG). Stress granules are non-membranous, cytoplasmic organelles that assemble in response to stress, and promote cell survival. Deregulation of SGs has been linked to several pathological conditions, including cancer, neurodegeneration, ischemia, and viral infections. SGs have been shown to promote tumor growth and metastasis, as well as contribute to mechanisms of chemo-resistance. Thus, these signaling hubs are emerging as a potential target in cancer. Methods: To induce SG formation, human pancreatic ductal adenocarcinoma (PDAC) cell lines were treated with sodium arsenite, thapsigargin, or exposed to hypoxic conditions. Immunoflourescent staining of core SG proteins was then conducted. To investigate cell cycle-driven mechanisms of SG heterogeneity, cell cycle stage was identified by expression of a Fluorescent ubiquitination-based cell cycle indicator (Fucci) lentiviral system. To investigate cPLA2’s role in SG formation, the chemical inhibitor Pyrrophenone, an shRNA targeting cPLA2, and an exogenous full-length or cleaved form of cPLA2, were used. In vivo, ES149 cells expressing Luciferase and the Fucci system were orthotopically implanted into C57BL/6 mice. Established tumors were treated every 3 days with Vehicle, Oxaliplatin, Pyrrophenone, or a combination of the latter. Tumor size was tracked via IVIS Spectrum. Fucci flourescence was used to identify cell cycle phase. SGs and cell death were assessed by immunoflourescent staining of tumors. Results: Cancer cells show a high degree of heterogeneity in their production of SGs. Cell cycle phase dictates SG heterogeneity. G2 phase cells have significantly higher levels of SG formation as compared cells in G1/S. This process is regulated by cleavage of the cPLA2 protein. Oxaliplatin administration increases G2 phase content in tumors. Co-treatment with Oxaliplatin and Pyrrophenone significantly decreases SG formation, increases cell death, and leads to tumor regressions in vivo. Conclusions: G2 phase cancer cells develop higher levels of SGs when exposed to stressful stimuli, as compared to G1/S phase counterparts. The cPLA2 protein is cleaved into a catalytically dead form that is present in G1/S phases, but absent in G2 phase. Oxaliplatin treatment in vivo leads to an increase in G2 phase tumor cells. When paired with cPLA2 inhibition, this results in increased cancer cell death and decreased tumor growth.

Firm life cycle and accrual quality

We examine the role of firms' life cycle stages on accrual quality (AQ). Our findings are consistent with an inverted U-shaped pattern in AQ across life cycle stages, where mature firms report the highest quality of accruals, followed by growth and shake-out firms, with firms in the introduction and decline stages reporting the lowest AQ. We further find that AQ worsens when firms move to the decline stage or when they move out of the mature stage to the shake-out and/or decline stage. Our findings are robust to several alternative specifications, subsample analyses, AQ measures, and firm life cycle proxies. We contribute to the literature on accrual quality by providing a broad view of how firm life cycle and accrual quality are intrinsically linked.

Probabilistic material flow analysis of released nano titanium dioxide in Mexico

Engineered Nanomaterials (ENMs) or products containing ENMs, known as nano-enabled products are commercialized globally by a large number of companies. Concern about the potential risks and negative impacts of releasing ENMs into the environment is under investigation. For this reason, methodologies to estimate the probable mass concentrations of ENMs released in different regions of the world have been developed. As a first attempt to estimate the probable mass flows of nanosized titanium dioxide (nano-TiO2) released in Mexico, we developed a Probabilistic Material Flow Analysis (PMFA) for 2015.The model describes probabilistic mass flows of released nano-TiO2 during the life cycle of sunscreens, coatings, ceramic, and other nano-enabled products, including the flows through the solid waste and wastewater management systems, as well as the transfer of nano-TiO2 to three environmental compartments (atmosphere, topsoil, and surface water). The PMFA incorporates the uncertainty related to the input data. We observed that the most significant nano-TiO2 flows occur to the surface water, landfill, and soil compartments, targeted as the main “hot-spots”, where living organisms could be more exposed to this material. Further improvements in the model are needed due to some data gaps at some life cycle stages, for instance, solid waste management and reused wastewater manipulation for irrigation purposes. Finally, the model developed in this study can be adjusted to assess other ENM releases and can be beneficial for further investigation in fate modeling and environmental risk assessment.

Evaluating novel building products through a building design-oriented LCA approach: Example of VIPs in terrace applications

Abstract INTRODUCTION: The construction industry is considered conservative in adopting new products/technologies, and environmental characteristics are one of the most important features of novel solutions. In this context, our study aims to present a building design-oriented assessment of a novel building product by extending the functional unit scope and involving multiple realistic building design scenarios. The task will be performed using the example of vacuum insulation panels (VIPs), a superinsulation product that can be used in various building applications. The study will focus on terrace insulation applications, where VIPs are an attractive solution for building designers due to the possibility of barrier-free floor design. However, they lack environmental evaluation. METHOD: A life cycle assessment (LCA) analysis is performed on two hypothetical buildings located in Ljubljana (Slovenia) for the cradle-to-gate and operational energy life cycle stages (A1-A3 + B6 according to EN 15978). A whole-year dynamic thermal response simulation was executed using the Design Builder software. Five barrier-free terrace design scenarios that influence the embodied and operational carbon footprint (while maintaining the functional and architectural integrity of the building design) were examined. RESULTS: The study showed that although EPS insulation showed a smaller embodied carbon footprint on the product level, the building level analysis showed that using VIPs in terrace applications leads to favourable or comparable environmental impact. CONCLUSIONS: Building products can be evaluated on three functional unit levels: product, application and building. By extending the boundaries from the product/application level to the entire building level, the study provides an example of a building design-oriented approach to LCA. The complexity increases by upgrading the functional unit scope to the building level, while the results become more case-specific and less general. Ideally, a novel building product should show environmental superiority on all three levels. However, as there are almost countless design possibilities in buildings, environmental superiority (or inferiority) on the product level does not necessarily indicate superiority (or inferiority) on the building level.

Unfolding the role of cell state on stress granule heterogeneity and function in pancreatic cancer.

e16357 Background: A major impediment to cancer therapy is the reality that cancer is a heterogeneous disease. A single tumor can consist of many genotypic or phenotypic profiles, which are marked by degrees of sensitivity to different therapeutics. Identifying and targeting origins of cancer cell heterogeneity is critical for predicting patient outcome and enhancing treatment efficacy. We observe a novel source of cancer cell heterogeneity that involves one of the largest stress-sensing platforms described to date: the stress granule (SG). Stress granules are non-membranous, cytoplasmic organelles that assemble in response to stress, and promote cell survival. Deregulation of SGs has been linked to several pathological conditions, including cancer, neurodegeneration, ischemia, and viral infections. SGs have been shown to promote tumor growth and metastasis, as well as contribute to mechanisms of chemo-resistance. Thus, these signaling hubs are emerging as a potential target in cancer. Methods: To induce SG formation, human pancreatic ductal adenocarcinoma (PDAC) cell lines were treated with sodium arsenite, thapsigargin, or exposed to hypoxic conditions. Immunoflourescent staining of core SG proteins was then conducted. To investigate cell cycle-driven mechanisms of SG heterogeneity, cell cycle stage was identified by expression of a Fluorescent ubiquitination-based cell cycle indicator (Fucci) lentiviral system. To investigate cPLA2’s role in SG formation, the chemical inhibitor Pyrrophenone, an shRNA targeting cPLA2, and an exogenous full-length or cleaved form of cPLA2, were used. In vivo, ES149 cells expressing Luciferase and the Fucci system were orthotopically implanted into C57BL/6 mice. Established tumors were treated every 3 days with Vehicle, Oxaliplatin, Pyrrophenone, or a combination of the latter. Tumor size was tracked via IVIS Spectrum. Fucci flourescence was used to identify cell cycle phase. SGs and cell death were assessed by immunoflourescent staining of tumors. Results: Cancer cells show a high degree of heterogeneity in their production of SGs. Cell cycle phase dictates SG heterogeneity. G2 phase cells have significantly higher levels of SG formation as compared cells in G1/S. This process is regulated by cleavage of the cPLA2 protein. Oxaliplatin administration increases G2 phase content in tumors. Co-treatment with Oxaliplatin and Pyrrophenone significantly decreases SG formation, increases cell death, and leads to tumor regressions in vivo. Conclusions: G2 phase cancer cells develop higher levels of SGs when exposed to stressful stimuli, as compared to G1/S phase counterparts. The cPLA2 protein is cleaved into a catalytically dead form that is present in G1/S phases, but absent in G2 phase. Oxaliplatin treatment in vivo leads to an increase in G2 phase tumor cells. When paired with cPLA2 inhibition, this results in increased cancer cell death and decreased tumor growth.

Identification and Root Cause Analysis of the Visible Particles Commonly Encountered in the Biopharmaceutical Industry.

Visible particle is an important issue in the biopharmaceutical industry, and it may occur across all the stages in the life cycle of biologics. Upon the occurrence of visible particles, it is often necessary to conduct chemical identification and root cause analysis to safeguard the safety and efficacy of the biotherapeutic products. In this article, we present a number of typical particles and relevant root cause analysis in the categories of extrinsic, intrinsic and inherent particles that are commonly encountered in the biopharma industry. In particular, the optical images of particles obtained both in situ and after isolation are provided, along with the spectral and elemental information. The particle identification was carried out with multiple microscopic and microspectroscopic techniques, including stereo optical microscopy, Fourier transform infrared microscopy, confocal Raman microscopy, scanning electron microscopy and energy dispersive X-ray spectroscopy. Both commercial and in-house spectral databases were used for comparison and identification. In addition to particle identification, our significant efforts are placed on the root cause analysis of the addressed particles with the intention to provide a relatively whole picture of the particle related issues and practical references to particle mitigation for our peers in the biopharmaceutical industry.

Assessing dengue risk globally using non-Markovian models

Dengue is a vector-borne disease transmitted by Aedes mosquitoes. The worldwide spread of these mosquitoes and the increasing disease burden have emphasized the need for a spatio-temporal risk map capable of assessing dengue outbreak conditions and quantifying the outbreak risk. Given that the life cycle of Aedes mosquitoes is strongly influenced by habitat temperature, numerous studies have utilized temperature-dependent development rates of these mosquitoes to construct virus transmission and outbreak risk models. In this study, we contribute to existing research by developing a mechanistic model for the mosquito life cycle that accurately captures its non-Markovian nature. Beginning with integral equations to track the mosquito population across different life cycle stages, we demonstrate how to derive the corresponding differential equations using phase-type distributions. This approach can be further applied to similar non-Markovian processes that are currently described with less accurate Markovian models. By fitting the model to data on human dengue cases, we estimate several model parameters, allowing the development of a global spatiotemporal dengue risk map. This risk model employs temperature and precipitation data to assess the environmental suitability for dengue outbreaks in a given area.

Технологическое обеспечение эксплуатационных свойств сопрягаемых поверхностей элементов сборной формообразующей оснастки с учетом установленной долговечности пресс-форм

Design and technological concepts are viewed at the stages of designing, manufacturing and joining process providing for the required reliability (durability) of mold elements. Recommendations are given for rationing and achieving the parameters of accuracy and quality of mating surfaces, taking into account the formation of the necessary operational properties at the main stages of the product life cycle. The factors that have the greatest impact on ensuring the correct and accurate location of forming tool elements have been elicited. The concept of determining the actual contact area and calculating the scale parameter, which provides a contact interface degree specification for conjugated surfaces, is presented. The hypothesis of the formation of reliability indicators is found, taking into account the manufacturing capability and technical capacity of production with the introduction of correction of the upper and lower dimensional deviations included in the dimension chains and ensuring the pre-set accuracy parameters of the master link. The analysis of the operating conditions of the studied type of pressmold elements is carried out. The main operational properties that provide the required operating time for shaping part failures are identified and analyzed. Special attention is paid to the study of the structural features of such parts, a comparison of the integral and prefabricated structures is made, and also, contact stresses under the joining operation for the most promising of the latter are calculated for contact interface degree simulation. This will allow developing practical recommendations for any type of compound forming tool for the benefit of basic mechanical properties rate making, accuracy indicators, including deviation allowances in the correct geometric shape, including quality parameters of the surface layer at the designing stage,processing methods dispatch and modes setting at the manufacturing and testing stages and also suggesting the amount of static loading when assembling.

The erosional and weathering response to arc–continent collision in New Guinea

Arc–continent collision is a fundamental stage in the plate-tectonic cycle that allows the continental crust to grow and can influence global climate through chemical weathering. Collision between Australia and the oceanic North Coast Range–New Britain Arc began in the Middle Miocene, resulting in uplift of the modern New Guinea Highlands. The temporal evolution of this collision and its erosional and weathering impacts is reconstructed here using sedimentary archives from the Gulf of Papua. Sr and Nd isotopes show dominant erosion from igneous arc-ophiolite crust, accounting for c . 40–70% of the total flux in the Early Miocene, and rising to c . 80–90% at 8 Ma, before falling again to 72–83% by the present day. Greater erosion from Australia-derived units accelerated in the Pliocene, like the classic Taiwan collision but with greater erosion from arc rather than continental units. Chemical alteration of the sediment increased through time, especially since c . 5 Ma, consistent with increasing kaolinite indicative of more tropical weathering. Erosion was focused in the high topography where mafic arc units are preferentially exposed. Comparison of sediment with bedrock compositions implies that the source terrains have been more efficient at removing CO 2 from the atmosphere compared with Himalayan drainages. Supplementary material : Details of analytical conditions, analytical results and compositions compiled from the GEOROC database are available at https://doi.org/10.6084/m9.figshare.c.7168147

Supporting building life cycle carbon monitoring, reporting and verification: A traceable and immutable blockchain-empowered information management system and application in Hong Kong

The Hong Kong Green Building Council introduced the Climate Change Framework for the Built Environment, aimed at enhancing the monitoring and management of life cycle carbon emissions within the building industry. In response to this initiative, this paper presents a novel blockchain-based carbon audit tool designed to facilitate the monitoring, reporting, and verification (MRV) of embodied carbon in Hong Kong's building sector. Specifically, a consortium blockchain is utilized for system development in this study. The proposed blockchain system records the information pertaining to each stage of the building's life cycle, which is uploaded for storage. The system ensures the traceability, immutability, and transparency of the recorded information. To test and verify the system, it was initially applied to manage and audit the embodied carbon in Hong Kong's building sector in 2020. We anticipate our findings will provide a viable tool for MRV of building life cycle carbon emissions.

Combined Effects of Annexin A5 Overexpression, 5-Fluorouracil Treatment, and Irradiation on Cell Viability of Caski Cervical Cancer Cell Line.

Cervical cancer is the fourth leading cause of cancer deaths in women globally. Combining gene therapy with chemo- and radiotherapy may improve cervical cancer treatment outcomes. This study evaluated the effects of Annexin A5(ANXA5) overexpression alongside 5-fluorouracil (5-FU) and irradiation on the viability of CaSki cervical squamous cell carcinoma (SCC) cells. pAdenoVator-CMV-ANXA5-IRES-GFP-plasmid and mock plasmid were transfected into CaSki cells using calcium-phosphate. Seventy-two hours post-transfection, GFP expression was quantified by fluorescence microscopy and flow cytometry to evaluate transfection efficiency. ANXA5 overexpression was confirmed via qPCR. Twenty-four hours post-transfection, cells received a single dose of 8Gy and were treated with 1 and 2µg/ml of 5-FU (IC50 = 2.783µg/ml). Cell viability, apoptosis, cell cycle stage, and Bcl-2 and Bax gene expression were assessed via MTT, annexin V/7-AAD, PI staining, and qPCR assays, respectively. ANXA5 was overexpressed 31.5-fold compared to control (p < 0.0001). MTT assays showed ANXA5 overexpression dose-dependently reduced CaSki cell viability (p < 0.001). IC50 of 5-FU was reduced from 2.783μg/mL to 1.794μg/mL when combined with ANXA5 overexpression. Additive effects on cell death were observed for ANXA5 plus 5-FU or irradiation versus ANXA5 alone. Apoptosis assays indicated combinatorial treatment increased CaSki cell apoptosis over ANXA5 alone. Cell cycle analysis revealed ANXA5 arrested cell cycle at G1/S phases; the percentage of cells in the S phase further rose with combination treatment. Finally, combination therapy significantly decreased Bcl-2 expression and increased Bax versus control (p < 0.001). Altogether, ANXA5 overexpression alongside 5-FU and irradiation may improve cervical squamous cell carcinoma (SCC) treatment efficacy. Further, in vivo investigations are warranted to confirm these in vitro results.

Sex- and cycle-dependent changes in spine density and size in hippocampal CA2 neurons

Sex hormones affect structural and functional plasticity in the rodent hippocampus. However, hormone levels not only differ between males and females, but also fluctuate across the female estrous cycle. While sex- and cycle-dependent differences in dendritic spine density and morphology have been found in the rodent CA1 region, but not in the CA3 or the dentate gyrus, comparable structural data on CA2, i.e. the hippocampal region involved in social recognition memory, is so far lacking. In this study, we, therefore, used wildtype male and female mice in diestrus or proestrus to analyze spines on dendritic segments from identified CA2 neurons. In basal stratum oriens, we found no differences in spine density, but a significant shift towards larger spine head areas in male mice compared to females. Conversely, in apical stratum radiatum diestrus females had a significantly higher spine density, and females in either cycle stage had a significant shift towards larger spine head areas as compared to males, with diestrus females showing the larger shift. Our results provide further evidence for the sexual dimorphism of hippocampal area CA2, and underscore the importance of considering not only the sex, but also the stage of the estrous cycle when interpreting morphological data.

EXPRESS: Self-Donations and Charitable Contributions in Online Crowdfunding: an Empirical Analysis

Many charitable projects have started using online crowdfunding platforms to raise donations. The rise of these platforms as fundraising vehicles has been partially driven by easy access to a large pool of potential donors without the significant marketing costs that commonly accompany traditional fundraising. However, such a low cost of entry also results in a significant "crowding" of projects, making it difficult for donors to decide which projects to donate to. Thus, a charitable project encounters a fundamental marketing challenge of standing out from other projects when conventional techniques like advertising and promotion are limited. In this paper, we posit that a project can credibly signal its quality via a strategy of "self-donation," whereby the project steward donates to her own project. Our empirical setting is an online education crowdfunding platform. By examining millions of donations, we find that self-donations improve the donation pace, contributed amount, and funding success. We show that the self-donation strategy works only when a self-donation is visible to potential donors and is especially effective at the early stage of the funding cycle or when project stewards are inexperienced, where the projects face significant uncertainty. We find evidence for self-donation as a quality signal through various observable proxies like impact letters to donors and corporate matching. Overall, our findings are consistent with a signaling mechanism that allows the separation of high-quality projects from lower-quality ones.