Age at first sexual intercourse, age at menarche, and age at menopause: a mendelian randomization study on lung cancer risk.

Background: There is increasing evidence that sex hormones are involved in the development of lung cancer, but the correlation between the reproductive behavior that changes sex hormone levels and lung cancer is not yet clear. Many previous studies have investigated the association between reproductive factors and lung cancer risk, but the results have been inconsistent. Therefore, we conducted a two-sample Mendelian randomization (MR) analysis to explore the potential relationship between age at first sexual intercourse (AFS), age at menarche, and age at menopause, and lung cancer. Methods: We performed a MR analysis of the data from the genome-wide association study (GWAS) of European ancestry to evaluate the independent effects of three reproductive behaviors on lung cancer overall (LUCA), lung adenocarcinoma (LUAD), lung squamous cell carcinoma (LUSC), and small cell lung cancer (SCLC). We mainly used the inverse-variance weighting method for the MR analysis. Sensitivity was determined by a MR-pleiotropy residual sum and outlier analysis, a weighted median analysis, a MR-Egger analysis, and a leave-one-out analysis. Results: The MR analysis results revealed that older AFS had a causal relationship with LUCA [odds ratio (OR) =0.6283, 95% confidence interval (CI): 0.4959-0.7961, P=0.0001), LUAD (OR =0.7042, 95% CI: 0.4967-0.9984, P=0.049), and LUSC (OR =0.6231, 95% CI: 0.4386-0.8853, P=0.0083). Conclusions: Our results revealed a causal relationship between older AFS and a lower risk of lung cancer. Our findings emphasize the importance of providing sex education, as early sexual intercourse may have undesirable effects. In addition, early psychological treatment is also essential.

Consensus for antifungal stewardship in China (2024 edition).

Background: Invasive fungal disease (IFD) has become a serious threat to human health in China and around the world, with high mortality and morbidity. Currently, the misdiagnosis rate of IFD is extremely high, compounded with the low quality of prescription antifungals and the high incidence of adverse events associated with IFD treatment, resulting in lengthy hospitalization, low clinical response, and high disease burden, which have become serious challenges in clinical practice. Antifungal stewardship (AFS) can not only significantly increase the early diagnosis rate of IFD, reduce inappropriate utilization of antifungal drugs, improve patient prognosis, but can also improve therapeutic safety and reduce healthcare expenses. Thus, it is urgent to identify key AFS metrics suitable for China's current situation. Methods: Based on metrics recommended by international AFS consensuses, combined with the current situation of China and the clinical experience of authoritative experts in various fields, several metrics were selected, and experts in the fields of respiratory diseases, hematology, intensive care units (ICUs), dermatology, infectious diseases, microbiology laboratory and pharmacy were invited to assess AFS metrics by the Delphi method. Consensus was considered to be reached with an agreement level of ≥80% for the metric. Results: Consensus was reached for 24 metrics, including right patient metrics (n=4), right time metrics (n=3), and right use metrics (n=17). Right use metrics were further subdivided into drug choice (n=8), drug dosage (n=4), drug de-escalation (n=1), drug duration (n=2), and drug consumption (n=2) metrics. Forty-six authoritative experts assessed and reviewed the above metrics, and a consensus was reached with a final agreement level of ≥80% for 22 metrics. Conclusions: This consensus is the first to propose a set of AFS metrics suitable for China, which helps to establish AFS standards in China and is also the first AFS consensus in Asia, and may improve the standard of clinical diagnosis and treatment of IFD, and guide hospitals to implement AFS, ultimately promoting the rational use of antifungal drugs and improving patient prognosis.

The Possible Role of Mycotoxins in the Pathogenesis of Endometrial Cancer.

Endometrial cancer is one of the most common cancer types among women. Many factors can contribute to the development of this disease, including environmental factors and, thus, eating habits. Our study aims to determine the levels of various mycotoxins and their metabolites in the blood serum and endometrial tissue samples of participants with previously proven endometrial cancer and to find possible contributions to cancer development. In the cohort clinical trial, 52 participants aged between 44 and 86 were studied. The participants were divided into two groups: patients or matched controls. All patients had previously histologically diagnosed endometrial cancer. The cancer patients were divided into low-grade endometrioid and low- plus high-grade endometrioid groups. Controls had no history of endometrial malignancy or premalignancy. Blood serum and endometrial tissue samples were obtained from all study patients. We compared the concentrations of total Aflatoxins (Afs), Deoxynivalenol (DON), Ochratoxin-A (OTA), T2-toxin and HT2 toxin (T2/HT2 toxin), Zearalenone (ZEN), alpha-Zearalenol (α-ZOL), and Fumonisin B1 (FB1) in the serum and endometrium between the different study groups. As a result, we can see a significant correlation between the higher levels of Afs and zearalenone and the presence of endometrial cancer. In the case of Afs, DON, OTA, T2/HT2 toxins, ZEN, and alpha-ZOL, we measured higher endometrial concentrations than in serum. Considering the effect of mycotoxins and eating habits on cancer development, our results might lead to further research exploring the relationship between certain mycotoxins and endometrium cancer.

Antifungal Stewardship in Invasive Fungal Infections, a Systematic Review.

INTRODUCTION: Invasive fungal infections (IFI) are a group of life-threatening diseases associated with significant morbidity, mortality and high healthcare costs. Some modern management programs known as AFS (antifungal stewardship programs) have now been developed. The purpose of this systematic review is to evaluate the different declinations of antifungal stewardship programs (AFPs). METHODS: Articles were systematically reviewed using the PRISMA checklist 2020. EMBASE and MEDLINE/PubMED were searched using the term "antifungal stewardship" (2012-2022 data) on 2 January 2023. Eligible studies were those that described an AFS and included an intervention, performance evaluation and outcome measures. RESULTS: A total of 22/796 studies were included. Approximately two-thirds (16) were published between 2018 and 2022. 16 (72.7%) stated a minimal complete AFS team. 12 (54.5%) adopted a non-compulsory AFS approach, 6(27.3%) had an Educational AFS and 4(18.2%) a compulsory AFS. Cost analyses of 12 studies showed a decrease for 7 (31.8%) compared to an increase for 5 (22.7%). In terms of outcomes, 18 studies showed a lower (10;45.5%) or the same (8;36.4%) pre-post intervention mortality rate. CONCLUSION: AFS programs seem to be related to lower costs and better outcomes and should thus be implemented in tandem with antimicrobial stewardship programs.

Terpene Synthase Gene Family in Chinese Chestnut (Castanea mollissima BL.) Harbors Two Sesquiterpene Synthase Genes Implicated in Defense against Gall Wasp Dryocosmus kuriphilus.

Chinese chestnut (Castanea mollissima BL.) is a well-known fruit tree that has been cultivated in East Asia for millennia. Leaves and buds of the plant can become seriously infested by the gall wasp Dryocosmus kuriphilus (GWDK), which results in gall formation and associated significant losses in fruit production. Herbivore-induced terpenes have been reported to play an important role in plant-herbivory interactions, and in this study, we show that upon herbivory by GWDK, four terpene-related compounds were significantly induced, while the concentrations of these four compounds in intact buds were relatively low. Among these compounds, (E)-nerolidol and (E, E)-α-farnesene have frequently been reported to be involved in plant herbivory defenses, which suggests direct and/or indirect functions in chestnut GWDK defenses. Candidate terpene synthase (TPS) genes that may account for (E)-nerolidol and (E, E)-α-farnesene terpene biosynthesis were characterized by transcriptomics and phylogenetic approaches, which revealed altered transcript levels for two TPSs: CmAFS, a TPS-g subfamily member, and CmNES/AFS, a TPS-b clade member. Both genes were dramatically upregulated in gene expression upon GWDK infestation. Furthermore, Agrobacterium tumefaciens-mediated transient overexpression in Nicotiana benthamiana showed that CmAFS catalyzed the formation of (E, E)-α-farnesene, while CmNES/AFS showed dual (E)-nerolidol and (E, E)-α-farnesene synthase activity. Biochemical assays of the recombinant CmAFS and CmNES/AFS proteins confirmed their catalytic activity in vitro, and the enzymatic products were consistent with two of the major volatile compounds released upon GWDK-infested chestnut buds. Subcellular localization demonstrated that CmAFS and CmNES/AFS were both localized in the cytoplasm, the primary compartment for sesquiterpene synthesis. In summary, we show that two novel sesquiterpene synthase genes CmAFS and CmNES/AFS are inducible by herbivory and can account for the elevated accumulation of (E, E)-α-farnesene and (E)-nerolidol upon GWDK infestation and may be implicated in chestnut defense against GWDK herbivores.

What is the impact of endometriosis and the AFS stage on cumulative pregnancy rates in IVF programs?

BACKGROUND: Endometriosis is commonly observed in infertile women and can be staged with regard to severity [e.g. according to the American Fertility Society (AFS) classification]. This condition can cause infertility through impaired oocyte quality, fertilization disorders, tubal lesions, adhesions, deep infiltration, and adenomyosis. Although women with endometriosis often turn to in vitro fertilization (IVF) programs, the literature data on IVF outcomes are sometimes contradictory (i.e. the same as in other etiologies of infertility, or worse). The objective of the present study was to assess and compare pregnancy rates in women with and without endometriosis and according to the endometriosis stage. METHODS: We retrospectively studied clinical and ongoing pregnancy rates in IVF and the cumulative pregnancy rates after frozen/thawed embryo transfers, in women without endometriosis (group A) or with endometriosis (group B). We further compared groups in which endometriosis was staged according to the revised AFS classification: stage 1/2 (group C), stage 3/4 (group D, without endometrioma), and endometrioma alone (group E). RESULTS: We documented 430 cycles in group A and 460 in group B (including 56 in group C, 88 in group D and 316 in group E). After fresh or frozen/thawed embryo transfers, the differences in ongoing pregnancy rates between groups A and B were not significant. However the cumulative rates per couple were significantly lower (p < 0.05) in group D. CONCLUSIONS: We recommend IVF for women with endometriosis because the pregnancy rates are similar to those observed for women with other types of infertility. This approach is in line with the international guidelines issued by assisted reproductive technology societies. These results again raise the question of whether surgical resection of deep infiltrating endometriosis (stage 3/4) should be recommended before admission to an IVF program. Trial registration This study was approved by an institutional review board (CPP Ouest VI, Brest, France): reference: B2020CE.43.

Cu-nanoparticles@ graphene nanocomposite: A robust and efficient nanocomposite for micro-solid phase extraction of trace aflatoxins in different foodstuffs.

Cu-nanoparticles-immobilized graphene (Cu@G) nanocomposite was fabricated in this study by reducing Cu(II) ions in the presence of graphene oxide using a simple chemical reduction step. Cu@G nanocomposite was applied as a sorbent for the SPE of four aflatoxins (AFs). A reusable syringe was filled with the fabricated nanocomposite and used as a sorbent for the micro-solid phase extraction of four AFs (AFB1, AFB2, AFG1, AFG2). The impact of different analytical factors was fully investigated and optimized. Excellent recoveries, ranging from 92.0 to 108.5 %, were detected when evaluating target AFs in samples of rice, maize, and pistachio. The LOD, LOQ, and linear ranges were attained under optimal circumstances in the ranges of 0.0062 µg kg-1, 0.0192 µg kg-1, and 0.0-20 µg kg-1, respectively. The discovered approach provided the dual benefits of a high enrichment capability of Cu-nanoparticles via AFs complexation and a huge porosity of graphene sheets.

Increased Cattle Feeding Precision from Automatic Feeding Systems: Considerations on Technology Spread and Farm Level Perceived Advantages in Italy.

Automation reduces the impact of farming on climate change and helps farmers adapt to its financial impact. Automatic feeding systems (AFSs) increase the ruminant's feeding precision and ease operators' workload. Such systems exist on a spectrum, requiring varied levels of operator support and installation complexity. A recent survey on farmers pointed out that those already running an AFS and those willing to buy one appreciate its ease of use, the resulting animal welfare, and the resulting overall benefits (increased production, farm profitability, and reduced feed waste). Whether technologically complex or not, studies have confirmed the benefits that farmers perceive to be underlying the remarkable reduction in the environmental impact of feeding operations (AFSs are electrically powered), the increase in animal welfare resulting from reduced conflicts around accessing the feed bunk, and the constant availability of fresh feed. However, their introduction should follow accurate and holistic structural and economic planning for existing and newly built facilities. The availability of public subsidies plays an essential role in pushing farmers to adopt the most modern digital technologies, whose benefits may even increase when farmers couple them with interconnected sensors to monitor animals' physiological states.

Agroforestry systems in the mid-hills of the north-western Himalaya: A sustainable pathway to improved soil health and climate resilience.

Identifying the new tree crop combinations plays an important function in transforming the low input agriculture into land units with high economic returns, increasing carbon (C) sink and nutrients storage capacity, and acting as a panacea to achieve Sustainability Development Goals (SDGs). The present study aims to evaluate various tree-crop combinations for (i) biomass production, (ii) carbon accumulation, and (iii) soil nutrient enrichment of traditional and commercially evolved eight agroforestry systems (AFSs), including agri-silvi-horticulture system, agri-silviculture system, silvi-pasture, fruit tree, fodder tree, bamboo, melia and poplar based AFSs with sole cropping system in the mid-hill zone of the north-western Himalaya. The results demonstrated that poplar based AFS accumulated a higher amount of biomass (130.87 Mg ha-1) and carbon (65.44 Mg ha-1) closely followed by melia-based AFS. The C stored in leaf litter was higher (0.66 Mg ha-1) in poplar-based AFS, however, soil C stock was maximum (114.69 Mg ha-1) under bamboo-based AFS. Overall, the Melia based AFS exhibited a higher rate of carbon dioxide mitigation (19.30 Mg ha-1 yr-1) and C-sequestration (5.26 Mg ha-1 yr-1) than other studied AFSs. Moreover, soil macro-nutrients (available N, P, K, S and Ca) were maximum under bamboo-based AFS, on the other hand, the fruit-based AFS had the higher concentrations of micro-nutrients i.e., Cu (3.05), Fe (31.10 mg g-1) and Mn (17.31 mg g-1). The soil microbial counts were higher in poplar-based AFS, whereas, the soil quality index improved significantly under bamboo based and fruit tree based AFSs. Hence, it can be concluded that the experimentally evolved AFSs represent an effective approach for boosting C-sequestration, soil fertility, regenerating the soil and sustainability of hill agriculture in the north-western Himalayas over traditional AFSs and sole cropping.

Effect of women's fertility and sexual development on epigenetic clock: Mendelian randomization study.

BACKGROUND AND OBJECTIVES: In observational studies, women's fertility and sexual development traits may have implications for DNA methylation patterns, and pregnancy-related risk factors can also affect maternal DNA methylation patterns. The aim of our study is to disentangle any potential causal associations between women's fertility and sexual development traits and epigenetic clocks, as well as to search for probable mediators by using the Mendelian randomization (MR) method. METHODS: Instrumental variables for exposures, mediators, and outcomes were adopted from genome-wide association studies data of European ancestry individuals. The potential causal relationship between women's fertility and sexual development traits and four epigenetic clocks were evaluated by inverse variance weighted method and verified by other two methods. Furthermore, we employed multivariable MR (MVMR) adjusting for hypertension, hyperglycemia, BMI changes, and insomnia. Then, combining the MVMR results and previous research, we performed two-step MR to explore the mediating effects of BMI, AFS, and AFB. Multiple sensitivity analyses were further performed to verify the robustness of our findings. RESULTS: Leveraging two-sample MR analysis, we observed statistically significant associations between earlier age at first birth (AFB) with a higher HannumAge, PhenoAge and GrimAge acceleration(ß = - 0.429, 95% CI [- 0.781 to - 0.077], p = 0.017 for HannumAge; ß = - 0.571, 95% CI [- 1.006 to - 0.136], p = 0.010 for PhenoAge, and ß = - 1.136, 95% CI [- 1.508 to - 0.765], p = 2.03E-09 for GrimAge respectively) and age at first sexual intercourse (AFS) with a higher HannumAge and GrimAge acceleration(ß = - 0.175, 95% CI [- 0.336 to - 0.014], p = 0.033 for HannumAge; ß = - 0.210, 95% CI [- 0.350 to - 0.070], p = 0.003 for GrimAge, respectively). Further analyses indicated that BMI, AFB and AFS played mediator roles in the path from women's fertility and sexual development traits to epigenetic aging. CONCLUSIONS: Our study suggested that AFS and AFB are associated with epigenetic aging. These findings may prove valuable in informing the development of prevention strategies and interventions targeted towards women's fertility and sexual development experiences and their relationship with epigenetic aging-related diseases.

Coordinated Optimal Control of AFS and DYC for Four-Wheel Independent Drive Electric Vehicles Based on MAS Model.

The problem that it is difficult to balance vehicle stability and economy at the same time under the starting steering condition of a four-wheel independent drive electric vehicle (4WIDEV) is addressed. In this paper, we propose a coordinated optimal control method of AFS and DYC for a four-wheel independent drive electric vehicle based on the MAS model. Firstly, the angular velocity of the transverse pendulum at the center of mass and the lateral deflection angle of the center of mass are decoupled by vector transformation, and the two-degree-of-freedom eight-input model of the vehicle is transformed into four two-degree-of-freedom two-input models, and the reduced-dimensional system is regarded as four agents. Based on the hardware connection structure and communication topology of the four-wheel independent drive electric vehicle, the reduced-dimensional model of 4WIDEV AFS and DYC coordinated optimal control is established based on graph theory. Secondly, the deviation of the vehicle transverse swing angular velocity and mass lateral deflection angle from their ideal values is oriented by combining sliding mode variable structure control (SMC) with distributed model predictive control (DMPC). A discrete dynamic sliding mode surface function is proposed for the ith agent to improve the robustness of the system in response to parameter variations and disturbances. Considering the stability and economy of the ith agent, an active front wheel steering and drive torque optimization control method based on SMC and DMPC is proposed for engineering applications. Finally, a hardware-in-the-loop (HIL) test bench is built for experimental verification, and the results show that the steering angle is in the range of 0-5°, and the proposed method effectively weighs the system dynamic performance, computational efficiency, and the economy of the whole vehicle. Compared with the conventional centralized control method, the torque-solving speed is improved by 32.33 times, and the electrical consumption of the wheel motor is reduced by 16.6%.

Exploring carbonate rock wettability across scales: Role of (bio)minerals.

HYPOTHESIS: The wettability of carbonate rocks is expected to be affected by the organic components of biominerals which are complex, nanostructured organo-mineral assemblages. Elucidating the nanoscale mechanisms driving the wettability of solid surfaces will enable a better understanding of the role of biominerals in the wetting properties of carbonate rocks to control various geological, environmental and industrial processes. EXPERIMENTS: Using Atomic Force Microscopy and Spectroscopy (AFM/AFS) we probed the wettability properties of carbonate rocks with different amounts of organic material. The adhesion properties of two types of limestones were determined in liquid environments at different length scales (nm to mm) using functionalized tips with different chemical groups to determine the extent of surface hydrophobic and hydrophilic organo-mineral interactions. FINDINGS: We observed homogeneous hydrophobic areas at length scales below < 5 µm. The origin of this hydrophobicity is linked to the presence of organics, whose amount and spatial distribution depend on the rock composition. Specifically, our results reveal that the biogenic vs non-biogenic origin of the mineral grains is the main rock property controlling the wettability of the solid surface. Overall, our methodology offers a multi-scale approach to unravel the role that organic moieties and biominerals play in controlling the wettability of rock-water interfaces.

Multicriteria spatial model to prioritize degraded areas for landscape restoration through agroforestry.

Reconciling the restoration of ecosystem services within agricultural landscapes is an effort that has been advancing within degraded areas restoration through agroforestry systems. However, to contribute to the effectiveness of these initiatives, it is essential to integrate landscape vulnerability and local demands to better highlight in which areas the implementation of agroforestry systems should be prioritized. Thus, we developed a spatial hierarchization methodology as a decision support tool as an active strategy for agroecosystem restoration. The proposed method constitutes a spatial indicator of priority areas to guide agroforestry interventions, including resource allocation and public policies for payment for environmental services. The methodology consists of Multicriteria Decision Analysis implemented in GIS software by combining input datasets based on biophysical conditions, environmental and socioeconomic aspects, that integrated promotes an assessment of the environment fragility, the pressures and responses to land use dynamic; a strategy for landscape restoration and conservation of the natural habitats, and multiple specific scenarios for decision making regarding the agricultural and the local actors demands. The output of the model provides the spatial distribution of areas suitable for the implementation of agroforestry systems, sorted into four priority levels (Low, Medium, High, and Extreme priority). The method is a promising tool proposal for territorial management and governance and subsidizes future research on the flows of ecosystem services.•Assessment of the environment fragility and the pressures and responses to land use dynamic.•Strategy for landscape restoration and conservation of remaining natural habitats.•Multiple specific scenarios for decision making regarding the agricultural and the local actors demands.

Nuclear receptor subfamily 1 group D member 1 suppresses the proliferation, migration of adventitial fibroblasts, and vascular intimal hyperplasia via mammalian target of rapamycin complex 1/ß-catenin pathway.

BACKGROUND: In-stent restenosis hardly limits the therapeutic effect of the percutaneous vascular intervention. Although the restenosis is significantly ameliorated after the application of new drug-eluting stents, the incidence of restenosis remains at a high level. OBJECTIVE: Vascular adventitial fibroblasts (AFs) play an important role in intimal hyperplasia and subsequent restenosis. The current study was aimed to investigate the role of nuclear receptor subfamily 1, group D, member 1 (NR1D1) in the vascular intimal hyperplasia. METHODS AND RESULTS: We observed increased expression of NR1D1 after the transduction of adenovirus carrying Nr1d1 gene (Ad-Nr1d1) in AFs. Ad-Nr1d1 transduction significantly reduced the numbers of total AFs, Ki-67-positive AFs, and the migration rate of AFs. NR1D1 overexpression decreased the expression level of ß-catenin and attenuated the phosphorylation of the effectors of mammalian target of rapamycin complex 1 (mTORC1), including mammalian target of rapamycin (mTOR) and 4E binding protein 1 (4EBP1). Restoration of ß-catenin by SKL2001 abolished the inhibitory effects of NR1D1 overexpression on the proliferation and migration of AFs. Surprisingly, the restoration of mTORC1 activity by insulin could also reverse the decreased expression of ß-catenin, attenuated proliferation, and migration in AFs induced by NR1D1 overexpression. In vivo, we found that SR9009 (an agonist of NR1D1) ameliorated the intimal hyperplasia at days 28 after injury of carotid artery. We further observed that SR9009 attenuated the increased Ki-67-positive AFs, an essential part of vascular restenosis at days 7 after injury to the carotid artery. CONCLUSION: These data suggest that NR1D1 inhibits intimal hyperplasia by suppressing the proliferation and migration of AFs in a mTORC1/ß-catenin-dependent manner.

Ultraviolet assisted liquid spray dielectric barrier discharge plasma-induced vapor generation for sensitive determination of arsenic by atomic fluorescence spectrometry.

In this study, a novel sensitive method for As determination by atomic fluorescence spectrometry was developed based on UV-assisted liquid spray dielectric barrier discharge (UV-LSDBD) plasma-induced vapor generation. It was found that prior-UV irradiation greatly facilitates As vapor generation in LSDBD likely because of the increased generation of active substances and the formation of As intermediates with UV irradiation. The experimental conditions affecting the UV and LSDBD processes (such as formic acid concentration, irradiation time, the flow rates of sample, argon and hydrogen) were optimized in detail. Under the optimum conditions, As signal measured by LSDBD can be enhanced by about 16 times with UV irradiation. Furthermore, UV-LSDBD also offers much better tolerance to coexisting ions. The limit of detection was calculated to be 0.13 µg L-1 for As, and the relative standard deviation of the repeated measurements was 3.2% (n = 7). The accuracy and effectiveness of this new method were further verified by the analysis of simulated natural water reference sample and real water samples. In this work, UV irradiation was utilized for the first time as an enhancement strategy for PIVG, which opens a new approach for developing green and efficient vapor generation methods.

Fatigue of an Aluminum Foam Sandwich Formed by Powder Metallurgy.

In this paper, an aluminum foam sandwich (AFS) was prepared by the rolling composite-powder metallurgy method, and its fatigue properties were studied. It was compared with an AFS made by the adhesive method to study its fatigue properties more directly. In this experiment, the fatigue performance was investigated by studying the microscopic interface, fatigue life, deflection curve, and failure mode. The results show that the fatigue life of an AFS with the rolling composite-powder metallurgy method is much longer than that with the adhesive method. The failure mode of an AFS made by the rolling composite-powder metallurgy method is shear failure, and that of an AFS made by the adhesive method is shear failure and interface debonding. An AFS with the rolling composite-powder metallurgy method has better fatigue properties. This paper also explored the fatigue damage model using the fatigue modulus method, and the polynomial fitting method has a higher fitting degree.

How to Quantify DNA Compaction by TFAM with Acoustic Force Spectroscopy and Total Internal Reflection Fluorescence Microscopy.

Mitochondrial transcription factor A (TFAM) plays a key role in the organization and compaction of the mitochondrial genome. However, there are only a few simple and accessible methods available to observe and quantify TFAM-dependent DNA compaction. Acoustic Force Spectroscopy (AFS) is a straightforward single-molecule force spectroscopy technique. It allows one to track many individual protein-DNA complexes in parallel and to quantify their mechanical properties. Total internal reflection fluorescence (TIRF) microscopy is a high-throughput single-molecule technique that permits the real-time visualization of the dynamics of TFAM on DNA, parameters inaccessible with classical biochemistry tools. Here we describe, in detail, how to set up, perform, and analyze AFS and TIRF measurements to study DNA compaction by TFAM.

Tracing and tracking filamentous structures across scales: A systematic review.

Filamentous structures are ubiquitous in nature, are studied in diverse scientific fields, and span vastly different spatial scales. Filamentous structures in biological systems fulfill different functions and often form dynamic networks that respond to perturbations. Therefore, characterizing the properties of filamentous structures and the networks they form is important to gain better understanding of systems level functions and dynamics. Filamentous structures are captured by various imaging technologies, and analysis of the resulting imaging data addresses two problems: (i) identification (tracing) of filamentous structures in a single snapshot and (ii) characterizing the dynamics (i.e., tracking) of filamentous structures over time. Therefore, considerable research efforts have been made in developing automated methods for tracing and tracking of filamentous structures. Here, we provide a systematic review in which we present, categorize, and discuss the state-of-the-art methods for tracing and tracking of filamentous structures in sparse and dense networks. We highlight the mathematical approaches, assumptions, and constraints particular for each method, allowing us to pinpoint outstanding challenges and offer perspectives for future research aimed at gaining better understanding of filamentous structures in biological systems.

Anterior fibromuscular stroma-preserved endoscopic enucleation of the prostate: a precision anatomical approach.

BACKGROUND: Benign prostate obstruction (BOO) is becoming increasingly important in this aging society. Some urge/stress urinary incontinence (UUI/SUI) still occurs after endoscopic enucleation of the prostate (EEP). It remains unclear how post-EEP incontinence can be avoided. Currently, early apical release to ameliorate the traction of the external sphincter is the best technique for incontinence prevention. OBJECTIVE: To describe our surgical technique of anterior fibromuscular stroma (AFS)-preserved EEP for BOO. DESIGN, SETTING, AND PARTICIPANTS: The medical records of 60 consecutive patients who underwent AFS-preserved EEP for BOO in our center from September 2019 to December 2019 were retrospectively reviewed. SURGICAL PROCEDURE: AFS-preserved EEP starts at the 12 o'clock position of the urethra, and the junction between the AFS and transitional zone (T-zone) was identified. The AFS and T-zone were separated first to protect the AFS in the initial operative procedure. Then, following the usual enucleation procedure, AFS-preserved EEP could be achieved. MEASUREMENTS: Postoperative prostate-specific antigen (PSA), testosterone, urethral stricture, and voiding status, such as incontinence, uroflow, and postvoiding residual urine were assessed. RESULTS AND LIMITATIONS: The data show that AFS-preserved EEP could achieve similar surgical outcomes as other early apical release approaches. CONCLUSIONS: The preserved AFS provides a nice landmark at the 12 o'clock position during EEP.

Sounding a New Era in Biomechanics with Acoustic Force Spectroscopy.

The acoustic force spectroscopy (AFS) tool was recently introduced as a novel tool for probing mechanical properties of biomolecules, expanding the application of sound waves to high-throughput quantification of the mechanical properties of single cells. By using controlled acoustic forces in the piconewton to nanonewton range, tens to hundreds of cells functionalized by attached microspheres can simultaneously be stretched and tracked in real-time with sub millisecond time response. Since its first application, several studies have demonstrated the potential and versatility of the AFS for high-throughput measurements of force-induced molecular mechanisms, revealing insight into cellular biomechanics and mechanobiology at the molecular level. In this chapter, we describe the operation of the AFS starting with the underlying physical principles, followed by a run-down of experimental considerations, and finally leading to applications in molecular and cellular biology.