Function of a complex of p-Y42 RhoA GTPase and pyruvate kinase M2 in EGF signaling pathway in glioma cells.

Epidermal growth factor (EGF) is known to be a critical stimulant for inducing the proliferation of glioma cancer cells. In our study, we observed that GST-RhoA binds to pyruvate kinase M2 (PKM2) in vitro. While EGF reduced the levels of RhoA protein, it significantly increased p-Y42 RhoA, as well as PKM1 and PKM2 in LN18 glioma cell line. We determined that RhoA undergoes degradation through ubiquitination involving SCF1 and Smurf1. Interestingly, we observed that p-Y42 RhoA binds to PKM2, while the dephosphomimetic form, RhoA Y42F, did not. Additionally, our observation revealed that PKM2 stabilized both RhoA and p-Y42 RhoA. Importantly, RhoA, p-Y42 RhoA, and PKM2, but not RhoA-GTP, were localized in the nucleus upon EGF stimulation. Knockdown of RhoA with siRNA resulted in the reduced levels of phosphoglycerate kinase1 (PGK1) and microtubule affinity-regulating kinase 4 (MARK). Furthermore, we found that the promoter of PGK1 was associated with ß-catenin and YAP. Notably, p-Y42 RhoA and PKM2 co-immunoprecipitated with ß-catenin and YAP. Based on these findings, we proposed a novel mechanism by which p-Y42 RhoA and PKM2, in conjunction with ß-catenin and YAP, regulate PGK1 expression, contributing to the progression of glioma upon EGF.

[Urinary incontinence after radical prostatectomy for prostate cancer-data from 17,149 patients from 125 certified centers]. / Harninkontinenz nach radikaler Prostatektomie beim Prostatakarzinom ­ aktuelle Daten von 17.149 Patienten aus 125 zertifizierten Zentren.

BACKGROUND: In addition to erectile dysfunction, urinary incontinence is the most common functional limitation after radical prostatectomy (RPE) for prostate cancer (PCa). The German S3 guideline recommends informing patients about possible effects of the therapy options, including incontinence. However, only little data on continence from routine care in German-speaking countries after RPE are currently available, which makes it difficult to inform patients. OBJECTIVE: The aim of this work is to present data on the frequency and severity of urinary incontinence after RPE from routine care. MATERIALS AND METHODS: Information from the PCO (Prostate Cancer Outcomes) study is used, which was collected between 2016 and 2022 in 125 German Cancer Society (DKG)-certified prostate cancer centers in 17,149 patients using the Expanded Prostate Cancer Index Composite Short Form (EPIC-26). Changes in the "incontinence" score before (T0) and 12 months after RPE (T1) and the proportion of patients who used pads, stratified by age and risk group, are reported. RESULTS: The average score for urinary incontinence (value range: 0-worst possible to 100-best possible) was 93 points at T0 and 73 points 12 months later. At T0, 97% of the patients did not use a pad, compared to 56% at T1. 43% of the patients who did not use a pad before surgery used at least one pad a day 12 months later, while 13% use two or more. The proportion of patients using pads differs by age and risk classification. CONCLUSION: The results provide a comprehensive insight into functional outcome 12 months after RPE and can be taken into account when informing patients.

The Complex of p-Tyr42 RhoA and p-p65/RelA in Response to LPS Regulates the Expression of Phosphoglycerate Kinase 1.

Inflammation plays a crucial role in tumorigenesis, primarily mediated by NF-κB. RhoA GTPases are instrumental in regulating the activation of NF-κB. Specifically, the phosphorylation of Tyrosine 42 on RhoA ensures the activation of NF-κB by directly activating the IKKß associated with IKKγ (NEMO). This study aimed to uncover the molecular mechanism through which p-Tyrosine 42 RhoA, in conjunction with NF-κB, promotes tumorigenesis. Notably, we observed that p-Tyrosine 42 RhoA co-immunoprecipitated with the p-Ser 536 p65/RelA subunit in NF-κB in response to LPS. Moreover, both p-Tyrosine 42 RhoA and p-p65/RelA translocated to the nucleus, where they formed a protein complex associated with the promoter of phosphoglycerate kinase 1 (PGK1) and regulated the expression of PGK1. In addition, p-p65/RelA and p-Tyr42 RhoA co-immunoprecipitated with p300 histone acetyltransferase. Intriguingly, PGK1 exhibited an interaction with ß-catenin, PKM1 and PKM2. Of particular interest, si-PGK1 led to a reduction in the levels of ß-catenin and phosphorylated pyruvate dehydrogenase A1 (p-PDHA1). We also found that PGK1 phosphorylated ß-catenin at the Thr551 and Ser552 residues. These findings discovered that PGK1 may play a role in transcriptional regulation, alongside other transcription factors.

Extracellular pyruvate kinase M2 induces cell migration through p-Tyr42 RhoA-mediated superoxide generation and epithelial-mesenchymal transition.

In the tumor microenvironment (TME), communication between cancer cells and tumor-associated macrophages (TAMs) through secreted extracellular proteins promotes cancer progression. Here, we observed that co-culturing cancer cells (4T1) and macrophage cells (Raw264.7) significantly enhanced superoxide production in both cell types. Using MALDI-TOF, we identified PKM2 as a highly secreted protein by Raw264.7 cells and bone marrow-derived monocytes. The extracellular recombinant PKM2 protein not only enhanced cancer cell migration and invasion but also increased superoxide production. Additionally, PKM2 was found to associate with the cell surface, and its binding to integrin α5/ß1 receptor was inhibited by antibodies specifically targeting it. Furthermore, we investigated downstream signaling pathways involved in PKM2-induced superoxide production. We found that knock-down of RhoA and p47phox using siRNAs effectively abolished superoxide generation in response to extracellular PKM2. Notably, extracellular PKM2 triggered the phosphorylation of p47phox at Ser345 residue and RhoA at Tyr42 residue (p-Tyr42 RhoA). Moreover, extracellular PKM2 exerted regulatory control over the expression of key epithelial-mesenchymal transition (EMT) markers, including ZEB1, Snail1, vimentin, and E-cadherin. Interestingly, p-Tyr42 RhoA translocated to the nucleus, where it bound to the ZEB1 promoter region. In light of these findings, we propose that extracellular PKM2 within the TME plays a critical role in tumorigenesis by promoting cancer cell migration and invasion through RhoA/p47phox signaling pathway.

Organic farming of maize crop enhances species evenness and diversity of hexapod predators.

Arthropod species diversity enhances ecosystem productivity and sustainability by increasing pollination and biological control services. Although, it is declining rapidly due to conventional agricultural intensification, organic agriculture with reduced reliance on agronomic inputs can regenerate ecosystems' resilience and restore them. Here, we report whether hexapod communities differ on both types of farming systems in small-scale field plot experiments, wherein Maize variety AG-589 was grown organically and conventionally in the 2020 and 2021 seasons. Livestock manure was applied in organic fields, whereas nitrogen and phosphorous were used as synthetic fertilizers in conventional fields. Hexapods were sampled three weeks after sowing once a week from the middle rows of subplots from both organically and conventionally grown maize. Twelve species of herbivores and four species of predators were recorded. Hexapod abundance overall and that of herbivores only was higher in conventionally cultivated maize, while predator abundance was higher in organic maize. Herbivores species diversity and evenness were significantly higher in conventional maize. Predator species diversity and evenness were significantly higher in organic maize fields. We noted predator abundance, diversity, and evenness as strong predictors to lower herbivore populations. These findings suggest that organic farming conserves natural enemies' biodiversity and regulates herbivores with increased provision of suitable habitats and prey resources for natural enemies, leading to enhanced relative abundance in their specialized niches. Thus, organic agriculture can potentially mediate better ecosystem services.

Deep Learning Based Multiresponse Optimization Methodology for Dual-Axis MEMS Accelerometer.

This paper presents a deep neural network (DNN) based design optimization methodology for dual-axis microelectromechanical systems (MEMS) capacitive accelerometer. The proposed methodology considers the geometric design parameters and operating conditions of the MEMS accelerometer as input parameters and allows to analyze the effect of the individual design parameters on the output responses of the sensor using a single model. Moreover, a DNN-based model allows to simultaneously optimize the multiple output responses of the MEMS accelerometers in an efficient manner. The efficiency of the proposed DNN-based optimization model is compared with the design of the computer experiments (DACE) based multiresponse optimization methodology presented in the Literature, which showed a better performance in terms of two output performance metrics, i.e., mean absolute error (MAE) and root mean squared error (RMSE).

[Primary pigmented malignant melanoma of the bladder treated with radical urethrocystectomy and adjuvant therapy]. / Primär pigmentiertes malignes Melanom in der Harnblase therapiert mit radikaler Urethrozystektomie und adjuvanter Therapie.

Primary malignant melanoma of the urinary tract is a rare entity with less than 50 cases reported worldwide. The following case concerns a 64-year-old woman whom we first saw in our emergency room due to gross haematuria. As part of the subsequent diagnostic investigation, we detected a primary malignant melanoma of the bladder and the urethra. The patient was treated with radical urethrocystectomy including pelvic lymphadenectomy and an ileum conduit. This was followed by a year of adjuvant therapy using checkpoint inhibitors.

New insights into the migration, distribution and accumulation of micro-plastic in marine environment: A critical mechanism review.

Microplastics (MPs) are widely distributed in the marine environment, posing a significant threat to marine biota. The contribution of anthropogenic and terrestrial sources to the aquatic ecosystem has led to an increase in MPs findings, and their abundance in aquatic biota has been reported to be of concern. MPs are formed mainly via photo degradation of macroplastics (large plastic debris), and their release into the environment is a result of the degradation of additives. Eco-toxicological risks are increasing for marine organisms, due to the ingestion of MPs, which cause damage to gastrointestinal (GI) tracts and stomach. Plastics with a size <5 mm are considered MPs, and they are commonly identified by Raman spectroscopy, Fourier transfer infrared (FTIR) spectroscopy, and Laser direct infrared (LDIR). The size, density and additives are the main factors influencing the abundance and bioavailability of MPs. The most abundant type of MPs found in fishes are fiber, polystyrenes, and fragments. These microscale pellets cause physiological stress and growth deformities by targeting the GI tracts of fishes and other biota. Approximately 80% MPs come from terrestrial sources, either primary, generated during different products such as skin care products, tires production and the use of MPs as carrier for pharmaceutical products, or secondary plastics, disposed of near coastal areas and water bodies. The issue of MPs and their potential effects on the marine ecosystem require proper attention. Therefore, this study conducted an extensive literature review on assessing MPs levels in fishes, sediments, seawater, their sources, and effects on marine biota (especially on fishes), chemo-physical behavior and the techniques used for their identification.

Lipopolysaccharide Stimulates A549 Cell Migration through p-Tyr 42 RhoA and Phospholipase D1 Activity.

Cell migration is a crucial contributor to metastasis, a critical process associated with the mortality of cancer patients. The initiation of metastasis is triggered by epithelial-mesenchymal transition (EMT), along with the changes in the expression of EMT marker proteins. Inflammation plays a significant role in carcinogenesis and metastasis. Lipopolysaccharide (LPS), a typical inflammatory agent, promoted the generation of superoxide through the activation of p-Tyr42 RhoA, Rho-dependent kinase 2 (ROCK2), and the phosphorylation of p47phox. In addition, p-Tyr42 RhoA activated phospholipase D1 (PLD1), with PLD1 and phosphatidic acid (PA) being involved in superoxide production. PA also regulated the expression of EMT proteins. Consequently, we have identified MHY9 (Myosin IIA, NMIIA) as a PA-binding protein in response to LPS. MYH9 also contributed to cell migration and the alteration in the expression of EMT marker proteins. Co-immunoprecipitation revealed the formation of a complex involving p-Tyr42 RhoA, PLD1, and MYH9. These proteins were found to be distributed in both the cytosol and nucleus. In addition, we have found that p-Tyr42 RhoA PLD1 and MYH9 associate with the ZEB1 promoter. The suppression of ZEB1 mRNA levels was achieved through the knockdown of RhoA, PLD1, and MYH9 using si-RNAs. Taken together, we propose that p-Tyr42 RhoA and PLD1, responsible for producing PA, and PA-bound MYH9 are involved in the regulation of ZEB1 expression, thereby promoting cell migration.

Multi-geostatistical analyses of the spatial distribution and source apportionment of potentially toxic elements in urban children's park soils in Pakistan: A risk assessment study.

In the past few decades, contamination of urban children's parks (UCPs) with potentially toxic elements (PTEs) has been attracting more and more interest; however, assessment of eco-environmental and child exposure risks particularly in developing countries remains limited. The current study investigated PTE (Cr, Ni, Zn, As, Cd, and Pb) concentrations, potential sources, and their health risk assessment in UCP soils of 12 major cities in Pakistan. The results showed that the mean concentration of Ni exceeded the SEPA-permissible limit in all UCP sites, while other PTEs were found to be within acceptable limits. The soil properties such as pH, electrical conductivity, organic matter, and soil particles size were determined in UCPs soils. The contamination factor and pollution load index results indicated low to moderate pollution levels (CF < 3) and (PLI<1) for all PTEs except Ni in some of the selected cities. Quantile-quantile (Q-Q) plotting determined the normal distribution line for all PTEs in the UCPs. Principal component analysis showed the mixed sources of contamination from industrial emissions, fossil fuel combustion, vehicular emissions, wastewater irrigation, as well as solid waste disposal and natural sources of soil parent materials in all park sites. ANOVA results showed that all the PTEs except Cd had moderate to higher contamination values than the reference site. The risk assessment study revealed that children had high exposure to the selected PTEs via all exposure pathways. The hazard index (HI) mean value (1.82E+00) of Ni for all exposure pathways was greater than 1, while total risk value of Cr (1.00E-03) had exceeded USEPA limit, indicating cancer risk. Consequently, the study of UCPs soils revealed PTEs contamination that could pose a potential health risk to the local population in the studied UCPs regions of Pakistan. Thus, the present study recommends that the influx of PTEs originating from natural and anthropogenic sources should be mitigated and government should implement strict enforcement of environmental regulations and proper management, as well as air quality monitoring guidelines for public health should be strictly adopted to reduce traffic- and industrial emission-related to PTEs in metropolitan areas.

Multiple functions of pyruvate kinase M2 in various cell types.

Glucose metabolism is a mechanism by which energy is produced in form of adenosine triphosphate (ATP) by mitochondria and precursor metabolites are supplied to enable the ultimate enrichment of mature metabolites in the cell. Recently, glycolytic enzymes have been shown to have unconventional but important functions. Among these enzymes, pyruvate kinase M2 (PKM2) plays several roles including having conventional metabolic enzyme activity, and also being a transcriptional regulator and a protein kinase. Compared with the closely related PKM1, PKM2 is highly expressed in cancer cells and embryos, whereas PKM1 is dominant in mature, differentiated cells. Posttranslational modifications such as phosphorylation and acetylation of PKM2 change its cellular functions. In particular, PKM2 can translocate to the nucleus, where it regulates the transcription of many target genes. It is notable that PKM2 also acts as a protein kinase to phosphorylate several substrate proteins. Besides cancer cells and embryonic cells, astrocytes also highly express PKM2, which is crucial for lactate production via expression of lactate dehydrogenase A (LDHA), while mature neurons predominantly express PKM1. The lactate produced in cancer cells promotes tumor progress and that in astrocytes can be supplied to neurons and may act as a major source for neuronal ATP energy production. Thereby, we propose that PKM2 along with its different posttranslational modifications has specific purposes for a variety of cell types, performing unique functions.

[The impact of using intraoperative photodynamic diagnosis on the prognosis of bladder cancer]. / Analyse der Anwendung der intraoperativen photodynamischen Diagnostik beim Harnblasenkarzinom.

BACKGROUND: Photodynamic Diagnosis (PDD) is widely used today for the diagnosis and treatment of superficial bladder cancer. This study aimed to analyse the impact of using this technique on recurrence rate, residual tumour rate and progression-free survival. MATERIALS AND METHODS: We conducted this study retrospectively in our unit between 10/2014 and 02/2018. We divided our patients into 2 groups: white light group, 49 patients who underwent primary bladder tumour resection with white light transurethral resection of bladder tumour (WL-TURB), and blue light group, 49 patients treated with blue light transurethral resection of bladder tumour (BL-TURB) with PDD. Patients in both groups were followed up for one year. We collected data for tumour recurrence for the first year, 3-month and 1-year relapse-free survival and 1-year progression-free assessment pursuant to the EORTC risk classification by Sylvester 2006. Furthermore, the residual tumour rate was determined by re-TURB. RESULTS: The following results were found independent of age, gender and histological findings: Residual tumour rate: blue light group 13.95 % vs. white light group 32.6 %, (p 0.02, OR 3.8, CI 95 % [1.17 - 12.85]).; 3-month relapse-free survival: blue light group 87.7 % vs. white light group 67.4 % (p 0.02, HR 3.18, CI 95 % [1.20 - 8.42]); 1-year relapse-free survival: blue light group 77.6 % vs. white light group 55.1 % (p 0.015, HR 2.56, CI 95 % [1.20 - 5.45]); 1-year progression-free survival: blue light group 95.9 % vs. white light group 79.6 %, p 0.03, HR 5.23, CI 95 % [1.11 - 24.53]) CONCLUSION: The use of hexaminolevulinate significantly reduces the risk of residual tumours compared with conventional TURB. The PDD technique significantly improved 3- and 12-month recurrence-free survival, especially in low and medium-risk tumours, and 1-year progression-free survival, especially in high-risk tumours.

[From stone treatment to coiling of aneurysm]. / Von der Steintherapie zum Aneurysmacoiling.

The following is a case report of a 54-year-old female who presented with persistent macrohaematuria with bladder tamponade three weeks after percutaneous nephrolithotomy. Angiography revealed an intrarenal aneurysm, which was closed by coiling, resulting in complete cessation of bleeding.

Impact of climate change on human health: evidence from riverine island dwellers of Bangladesh.

This study aims to explore the impact of climate change on health, including local adaptation strategies. A mixed-method approach has been used in this study. The results reveal that increasing the frequency of flooding, severity of riverbank erosion and drought, and rising disease outbreak are the highest indicators of climate change perceived by riverine island (char) dwellers, which is similar to the observed data. It also uncovers, approximately all respondents encounter several health-related issues during different seasons where prevailing cold and cough with fever, skin diseases, and diarrhoea are the leading ailments. Several adaptation strategies are accommodated by char inhabitants in order to enhance resilience against the climate change health impacts, but the paucity of money, disrupted communication, lack of formal health-care centre are the most obstacles to the sustainability of adaptation. This research recommends that healthcare-associated project should be performed through proper monitoring for exterminating char dwellers' health issues.

A Systematic Design Optimization Approach for Multiphysics MEMS Devices Based on Combined Computer Experiments and Gaussian Process Modelling.

This paper presents a systematic and efficient design approach for the two degree-of-freedom (2-DoF) capacitive microelectromechanical systems (MEMS) accelerometer by using combined design and analysis of computer experiments (DACE) and Gaussian process (GP) modelling. Multiple output responses of the MEMS accelerometer including natural frequency, proof mass displacement, pull-in voltage, capacitance change, and Brownian noise equivalent acceleration (BNEA) are optimized simultaneously with respect to the geometric design parameters, environmental conditions, and microfabrication process constraints. The sampling design space is created using DACE based Latin hypercube sampling (LHS) technique and corresponding output responses are obtained using multiphysics coupled field electro-thermal-structural interaction based finite element method (FEM) simulations. The metamodels for the individual output responses are obtained using statistical GP analysis. The developed metamodels not only allowed to analyze the effect of individual design parameters on an output response, but to also study the interaction of the design parameters. An objective function, considering the performance requirements of the MEMS accelerometer, is defined and simultaneous multi-objective optimization of the output responses, with respect to the design parameters, is carried out by using a combined gradient descent algorithm and desirability function approach. The accuracy of the optimization prediction is validated using FEM simulations. The behavioral model of the final optimized MEMS accelerometer design is integrated with the readout electronics in the simulation environment and voltage sensitivity is obtained. The results show that the combined DACE and GP based design methodology can be an efficient technique for the design space exploration and optimization of multiphysics MEMS devices at the design phase of their development cycle.

A Low-g MEMS Accelerometer with High Sensitivity, Low Nonlinearity and Large Dynamic Range Based on Mode-Localization of 3-DoF Weakly Coupled Resonators.

This paper presents a new design of microelectromechanical systems (MEMS) based low-g accelerometer utilizing mode-localization effect in the three degree-of-freedom (3-DoF) weakly coupled MEMS resonators. Two sets of the 3-DoF mechanically coupled resonators are used on either side of the single proof mass and difference in the amplitude ratio of two resonator sets is considered as an output metric for the input acceleration measurement. The proof mass is electrostatically coupled to the perturbation resonators and for the sensitivity and input dynamic range tuning of MEMS accelerometer, electrostatic electrodes are used with each resonator in two sets of 3-DoF coupled resonators. The MEMS accelerometer is designed considering the foundry process constraints of silicon-on-insulator multi-user MEMS processes (SOIMUMPs). The performance of the MEMS accelerometer is analyzed through finite-element-method (FEM) based simulations. The sensitivity of the MEMS accelerometer in terms of amplitude ratio difference is obtained as 10.61/g for an input acceleration range of ±2 g with thermomechanical noise based resolution of 0.22 µµg/Hz and nonlinearity less than 0.5%.

[Infected urachal cyst in an adult manifests as urosepsis]. / Infizierte Urachuszyste beim Erwachsenen, Manifestation als Urosepsis.

The following is a case of an 81-year-old male adult who presented to our emergency department with signs of urosepsis. The patient came with an indwelling urinary catheter due to benign prostatic hyperplasia. Extended diagnostic investigation demonstrated an infected urachal cyst as the cause of infection, which was surgically treated.

Microfabrication Process-Driven Design, FEM Analysis and System Modeling of 3-DoF Drive Mode and 2-DoF Sense Mode Thermally Stable Non-Resonant MEMS Gyroscope.

This paper presents microfabrication process-driven design of a multi-degree of freedom (multi-DoF) non-resonant electrostatic microelectromechanical systems (MEMS) gyroscope by considering the design constraints of commercially available low-cost and widely-used silicon-on-insulator multi-user MEMS processes (SOIMUMPs), with silicon as a structural material. The proposed design consists of a 3-DoF drive mode oscillator with the concept of addition of a collider mass which transmits energy from the drive mass to the passive sense mass. In the sense direction, 2-DoF sense mode oscillator is used to achieve dynamically-amplified displacement in the sense mass. A detailed analytical model for the dynamic response of MEMS gyroscope is presented and performance characteristics are validated through finite element method (FEM)-based simulations. The effect of operating air pressure and temperature variations on the air damping and resulting dynamic response is analyzed. The thermal stability of the design and corresponding effect on the mechanical and capacitive sensitivity, for an operating temperature range of -40 °C to 100 °C, is presented. The results showed that the proposed design is thermally stable, robust to environmental variations, and process tolerances with a wide operational bandwidth and high sensitivity. Moreover, a system-level model of the proposed gyroscope and its integration with the sensor electronics is presented to estimate the voltage sensitivity under the constraints of the readout electronic circuit.

P-Tyr42 RhoA GTPase amplifies superoxide formation through p47phox, phosphorylated by ROCK.

Optimal levels of reactive oxygen species (ROS) play a critical role in cellular physiological function. For production of intracellular superoxide, NADPH oxidase is one of the sources. Rac1/2 and RhoA GTPases are involved in regulation of NADPH oxidase activity and Tyr42 phosphorylation of RhoA (p-Tyr42 RhoA) seems significant in this regard as it was recently shown that hydrogen peroxide was able to increase p-Tyr42 RhoA levels. Phorbol myristate acetate (PMA), a tumor promoter, also induces production of superoxides; PMA activates Src, a tyrosine kinase, and increases p-Tyr42 RhoA levels. In exploring the mechanism of PMA effects, we reduced RhoA levels in test cells with si-RhoA and then restoration of various versions of RhoA for effect in response of the cells to PMA and producing superoxides. Restoration of RhoA Y42F (a dephospho-mimic form) still had reduced superoxide formation in response to PMA, compared with WT and Y42E RhoA. This was similarly seen with assays for cell migration and proliferation with cells responding to PMA. Y27632, a ROCK (Rho associated coiled coil kinase) inhibitor, also inhibited superoxide production, and also reduced p-Y416 Src and p-p47phox levels. A ROCK active fragment was also able to phosphorylate p47phox at Ser345 residue (p-Ser345 p47phox), a component of NADPH oxidase. Overall, we demonstrate that p-Tyr42 RhoA levels increase following PMA treatment and this is through production of superoxide and activation of Src. These in turn amplify superoxide production through ROCK phophorylation of p47phox and maintain a positive feedback loop for superoxide generation, and contribute to tumor progression.