Gain-of-Function Mutations in p53 in Cancer Invasiveness and Metastasis.

Forty years of research has proven beyond any doubt that p53 is a key regulator of many aspects of cellular physiology. It is best known for its tumor suppressor function, but it is also a regulator of processes important for maintenance of homeostasis and stress response. Its activity is generally antiproliferative and when the cell is damaged beyond repair or intensely stressed the p53 protein contributes to apoptosis. Given its key role in preventing cancer it is no wonder that it is the most frequently mutated gene in human cancer. Surprisingly, a subset of missense mutations occurring in p53 (gain-of-function) cause it to lose its suppressor activity and acquire new functionalities that turn the tumor suppressor protein into an oncoprotein. A solid body of evidence exists demonstrating increased malignancy of cancers with mutated p53 in all aspects considered "hallmarks of cancer". In this review, we summarize current findings concerning the cellular processes altered by gain-of-function mutations in p53 and their influence on cancer invasiveness and metastasis. We also present the variety of molecular mechanisms regulating these processes, including microRNA, direct transcriptional regulation, protein-protein interactions, and more.

Towards the Properties of Different Biomass-Derived Proteins via Various Extraction Methods.

This study selected three representative protein-rich biomass-brewer's spent grain (BSG), pasture grass (PG), and cyanobacteria (Arthrospira platensis; AP) for protein extraction with different extraction methods (alkaline treatment, aqueous extraction, and subcritical water extraction). The yield, purity, molecular weight, oil-water interfacial tension, and thermal stability of the obtained proteins derived from different biomass and extraction methods were comprehensively characterized and compared. In the view of protein yield and purity, alkaline treatment was found optimal for BSG (21.4 and 60.2 wt.%, respectively) and AP (55.5 and 68.8 wt.%, respectively). With the decreased oil-water interfacial tension, the proteins from all biomass showed the potential to be emulsifier. BSG and AP protein obtained with chemical treatment presented excellent thermal stability. As a novel method, subcritical water extraction is promising in recovering protein from all three biomass with the comparable yield and purity as alkaline treatment. Furthermore, the hydrolyzed protein with lower molecular weight by subcritical water could promote its functions of foaming and emulsifying.

The clathrin-binding and J-domains of GAK support the uncoating and chaperoning of clathrin by Hsc70 in the brain.

Cyclin-G-associated kinase (GAK), the ubiquitously expressed J-domain protein, is essential for the chaperoning and uncoating of clathrin that is mediated by Hsc70 (also known as HSPA8). Adjacent to the C-terminal J-domain that binds to Hsc70, GAK has a clathrin-binding domain that is linked to an N-terminal kinase domain through a PTEN-like domain. Knocking out GAK in fibroblasts caused inhibition of clathrin-dependent trafficking, which was rescued by expressing a 62-kDa fragment of GAK, comprising just the clathrin-binding and J-domains. Expressing this fragment as a transgene in mice rescued the lethality and the histological defects caused by knocking out GAK in the liver or in the brain. Furthermore, when both GAK and auxilin (also known as DNAJC6), the neuronal-specific homolog of GAK, were knocked out in the brain, mice expressing the 62-kDa GAK fragment were viable, lived a normal life-span and had no major behavior abnormalities. However, these mice were about half the size of wild-type mice. Therefore, the PTEN-like domains of GAK and auxilin are not essential for Hsc70-dependent chaperoning and uncoating of clathrin, but depending on the tissue, these domains appear to increase the efficiency of these co-chaperones.

Unbiased screen for interactors of leucine-rich repeat kinase 2 supports a common pathway for sporadic and familial Parkinson disease.

Mutations in leucine-rich repeat kinase 2 (LRRK2) cause inherited Parkinson disease (PD), and common variants around LRRK2 are a risk factor for sporadic PD. Using protein-protein interaction arrays, we identified BCL2-associated athanogene 5, Rab7L1 (RAB7, member RAS oncogene family-like 1), and Cyclin-G-associated kinase as binding partners of LRRK2. The latter two genes are candidate genes for risk for sporadic PD identified by genome-wide association studies. These proteins form a complex that promotes clearance of Golgi-derived vesicles through the autophagy-lysosome system both in vitro and in vivo. We propose that three different genes for PD have a common biological function. More generally, data integration from multiple unbiased screens can provide insight into human disease mechanisms.

Disruption of clathrin-mediated trafficking causes centrosome overduplication and senescence.

The Hsc70 cochaperone, G cyclin-associated kinase (GAK), has been shown to be essential for the chaperoning of clathrin by Hsc70 in the cell. In this study, we used conditional GAK knockout mouse embryonic fibroblasts (MEFs) to determine the effect of completely inhibiting clathrin-dependent trafficking on the cell cycle. After GAK was knocked out, the cells developed the unusual phenotype of having multiple centrosomes, but at the same time failed to divide and ultimately became senescent. To explain this phenotype, we examined the signaling profile and found that mitogenic stimulation of the GAK KO cells and the control cells were similar except for increased phosphorylation of Akt. In addition, the disruption of intracellular trafficking caused by knocking out GAK destabilized the lysosomal membranes, resulting in DNA damage due to iron leakage. Knocking down clathrin heavy chain or inhibiting dynamin largely reproduced the GAK KO phenotype, but inhibiting only clathrin-mediated endocytosis by knocking down adaptor protein (AP2) caused growth arrest and centrosome overduplication, but no DNA damage or senescence. We conclude that disruption of clathrin-dependent trafficking induces senescence accompanied by centrosome overduplication because of a combination of DNA damage and changes in mitogenic signaling that uncouples centrosomal duplication from DNA replication.

Influence of protocol variables on outcomes of the star excursion balance test group (SEBT, mSEBT, YBT-LQ) in healthy individuals: a systematic review.

Background: The "SEBT group," which includes the Star Excursion Balance Test (SEBT), its modified version (mSEBT), and the Lower Quarter Y-Balance Test (YBT-LQ), is used to assess the limits of stability. Interestingly, the testing protocol allows users a considerable degree of flexibility, which can affect the obtained results. Therefore, the objective of this systematic review was to analyze the impact of different protocol variants within the "SEBT group" on outcomes. Methods: Data were acquired by searching 4 databases (MEDLINE, ScienceDirect, Wiley, Springer Link) focusing on studies published in English in peer-reviewed journals, empirical in nature, conducted on healthy individuals, and examining the effects of various protocol variants on test outcomes. Study quality was assessed with the NHLBI quality assessment tool for pre-post studies with no control group. Results: The calculation method based on the maximum repetition yields statistically significantly higher results compared to other calculation methods. Allowing unrestricted arm movements during the test results in statistically significantly higher scores compared to the procedure that restricts arm movements. The impact of a warm-up, wearing footwear during testing, and using a dedicated kit remains ambiguous. To obtain reliable results, 4-6 familiarization trials are necessary, though fewer may suffice for athletes experienced in performing the test. Conclusion: This systematic review highlights the significant impact of the calculation method and arm movement restrictions on the outcomes of the "SEBT group." The effects of wearing footwear during testing, warm-up, and using a dedicated test kit remain unclear. The required number of familiarization repetitions may varies depending on biological maturity level of the person being tested. Future research should develop a warm-up protocol tailored to the needs of the "SEBT group," and investigate the impact of heel elevation during testing on outcomes. Systematic review registration: The protocol for this systematic review was prospectively registered in the OSF Registries (https://doi.org/10.17605/OSF.IO/JSKH2).

Ankle dorsiflexion range of motion and hip abductor strength can predict Lower Quarter Y-Balance Test performance in healthy males.

OBJECTIVES: The aim of the study was to assess whether strength and range of motion (ROM) of the hip and ankle are the factors determining performance in the Lower Quarter Y-Balance test (YBT-LQ). DESIGN: Cross-sectional study. PARTICIPANTS: 66 healthy males (age: 25.2±6.8 years) participated in this study. MAIN OUTCOME MEASURES: Participants underwent assessments of ankle dorsiflexion (DF) ROM, hip internal rotation (IR) ROM, external rotation (ER) ROM and isometric strength of hip abductor (ABD), extensor (EXT) and external rotators (ERS) muscles together with YBT-LQ for both legs. A forward 2-steps multiple linear regression analysis was conducted to examine the relationship between the predictor variables and the criterion variable. RESULTS: Ankle DF ROM predicted anterior (ANT) reach (R2 = 0.49; R2 = 0.33; p < 0.001). The model with hip ABD strength and ankle DF ROM explained posteromedial (PM) reach variance for stance leg (R2 = 0.35; p < 0.001), while only hip ABD strength was included for kicking leg (R2 = 0.19; p = 0.007). The model with ankle DF ROM and hip ABD strength explained posterolateral (PL) reach for stance leg (R2 = 0.41; p < 0.001). Hip ABD was the only predictor for kicking leg PL reach (R2 = 0.15; p < 0.001). YBT-LQ composite score was explained by ankle DF ROM and hip ABD strength for both legs (R2 = 0.44; p < 0.001) and (R2 = 0.25; p = 0.002). CONCLUSION: Hip ABD strength and ankle DF ROM can determine performance in the YBT-LQ. Strength of hip EXT, ERS as well as ROM of hip IR and ER did not predict YBT-LQ performance.

Advantages of Robotic Surgery for Patients of Reproductive Age with Endometrial Cancer.

This review presents current knowledge on the surgical treatment of endometrial cancer in young patients. Endometrial cancer is the most common gynecological cancer in Europe. Higher morbidity is correlated with obesity, hypertension and diabetes, which are growing worldwide. However, endometrial cancer at an early age is very rare. The first line of treatment for this cancer is radical hysterectomy, which is controversial in young women. There is an alternative method of fertility-sparing treatment. However, there is a group of young patients for whom surgical treatment is recommended. According to European guidelines, minimally invasive surgery is recommended for endometrial cancer. The aim of the study was to present the advantages of robotic surgery for endometrial cancer detected at a young age. The procedure of radical treatment with robot-assisted laparoscopy is more precise. Better visualization and stabilization of instruments allow a shorter procedure time, a brief hospital stay and fewer complications. Quality of life may be at a similar level. Incisions after trocars are painless and more esthetic than a classical wound. Bilateral adnexectomy in endometrial cancer depends on age, molecular status of the cancer, stage, genetic risk factors and individual decision. Conclusions: Robotic surgery seems to be a better surgical method for endometrial cancer in younger patients.

Circulating microRNAs from plasma as preclinical biomarkers of epileptogenesis and epilepsy.

Epilepsy frequently develops as a result of brain insult; however, there are no tools allowing to predict which patients suffering from trauma will eventually develop epilepsy. microRNAs are interesting candidates for biomarkers, as several of them have been described to change their levels in the brains, and in the plasma of epileptic subjects. This study was conducted to evaluate the usefulness of plasma miRNAs as epileptogenesis/epilepsy biomarkers. In our studies, we used a rat model of temporal lobe epilepsy. An epileptogenic insult was status epilepticus evoked by stimulation of the left lateral nucleus of the amygdala. Next, animals were continuously video and EEG monitored for 3 months. Blood was collected at 14, 30, 60, and 90 days after stimulation. Blood plasma was separated and miRNA levels were analyzed. We compared miRNA levels between sham-operated and stimulated animals, and between animals with high and low numbers of seizures. We propose three miRNAs that could be biomarkers of epilepsy: miR-671, miR-9a-3p and miR-7a-5p. According to us, miR-206-5p is a potential biomarker of epileptogenesis, and miR-221-3p is a potential biomarker of epilepsy severity. We think that these five miRNAs can be considered in the future as potential treatment targets.

Robotic surgery in endometrial cancer: first Polish experience.

In Poland, robotic surgery is nowadays perceived as a new method of surgical treatment in endometrial cancer patients. We aim to present the first Polish group of endometrial cancer patients treated using robotic surgery. The analysis was based on 79 patients with mean age of 59.72 ± 11.709 (range 27-83) years and endometrial cancer scheduled for surgical treatment. Mean BMI was 31.38 ± 8.78 (range 19.03-65.97) kg/m2. The data were collected based on a questionnaire consisting of 19 questions concerning the patient's perception of robotic surgery before the procedure. Patients with a family history of neoplastic diseases indicate precision of movements as the most important reason for choosing robotic surgery (p = 0.0035). Patients after surgery procedures in the past named shorter hospitalization as a major benefit (p = 0.0037). Patients who chose robotic surgery for financial reasons stressed the cosmetic effect as a priority (p = 0.0319). Shorter length of hospital stay, less blood loss, enlarged view, and good visualization were statistically significant reasons for choosing robotic surgery (p < 0.05). Women who consider work, good material status, and well-being as the most important aspects of their lives cited the cosmetic effect as a benefit of robotic surgery (p = 0.0029 vs. p = 0.0074 vs. p = 0.01745, respectively). In the follow-up after operations, no patients regretted choosing robotic surgery. Good visualization, precise movements, less blood loss, and cosmetic effects are the most frequent reasons for choosing robotic surgery. Even patients after other types of surgery in the past decided on robot-assisted radical hysterectomy because of the clear benefits of this approach.

Efficacy of Single- and Dual-Docking Robotic Surgery of Paraaortic and Pelvic Lymphadenectomy in High-Risk Endometrial Cancer.

(1) The surgical method of choice for the treatment of endometrial cancer is minimally invasive surgery. In cases of high-risk endometrial cancer, completed paraaortic and pelvic lymphadenectomy are indicated. The aim of this study was to analyze the types of docking during robotic surgery assisted with the da Vinci X system while performing paraaortic and pelvic lymphadenectomy. (2) Methods: A total of 25 patients with high-risk endometrial cancer, with a mean age of 60.07 ± 10.67 (range 34.69-83.23) years, and with a mean body mass index (BMI) of 28.4 ± 5.62 (range 18-41.5) kg/m2, were included in this study. The analyzed population was divided into groups that underwent single or dual docking during surgery. (3) Results: No statistical significance was observed between single and dual docking during paraaortic and pelvic lymphadenectomy and between the type of docking and the duration of the operation. However, there was a statistically significant correlation between the duration of the operation and previous surgery (p < 0.005). The number of removed lymph nodes was statistically associated with BMI (p < 0.005): 15.87 ± 6.83 and 24.5 ± 8.7 for paraaortic and pelvic lymph nodes, respectively, in cases of single docking, and 18.05 ± 7.92 and 24.88 ± 11.75 for paraaortic and pelvic lymph nodes, respectively, in cases of dual docking. (4) Conclusions: The robot-assisted approach is a good surgical method for lymphadenectomy for obese patients, and, despite the type of docking, there are no differences in the quality of surgery.

Antipsychotic compounds differentially modulate high-frequency oscillations in the rat nucleus accumbens: a comparison of first- and second-generation drugs.

Improved understanding of the actions of antipsychotic compounds is critical for a better treatment of schizophrenia. Abnormal oscillatory activity has been found in schizophrenia and in rat models of the disease. N-Methyl-D-aspartic acid receptor (NMDAR) antagonists, used to model certain features of schizophrenia, increase the frequency and power of high-frequency oscillations (HFO, 130-180 Hz) in the rat nucleus accumbens, a brain region implicated in schizophrenia pathology. Antipsychotics can be classified as first- and second-generation drugs, the latter often reported to have wider benefit in humans and experimental models. This prompted the authors to examine the pre- and post-treatment effects of clozapine, risperidone (second-generation drugs) and sulpiride and haloperidol (first-generation drugs) on ketamine and MK801-enhanced accumbal HFO. Both NMDAR antagonists increased HFO frequency. In contrast, clozapine and risperidone markedly and dose-dependently reduced the frequency of spontaneous and NMDAR-antagonist-enhanced HFO, whilst a moderate effect was found for sulpiride and a much weaker effect for haloperidol. Unexpectedly, we found reductions in HFO frequency were associated with an increase in its power. These findings indicate that modulation of accumbal HFO frequency may be a fundamental effect produced by antipsychotic compounds. Of the drugs investigated, first- and second-generation compounds could be dissociated by their potency on this measure. This effect may partially explain the differences in the clinical profile of these drugs.

Cross cultural adaptation, reliability and validity of the Polish version of the Cumberland Ankle Instability Tool.

Purpose: The aim of this study was to develop a Polish version of the Cumberland Ankle Instability Tool (CAIT-PL) questionnaire and perform CAIT-PL cultural adaptation with assessment of its psychometric properties.Methods: The original CAIT was translated into the Polish language using standard guidelines. The CAIT-PL was completed on 2 occasions by 105 participants both with and without a history of ankle sprains. Psychometric evaluation of CAIT-PL included an assessment of internal consistency, test-retest reliability, standard error of measurement, convergent and construct validity.Results: The CAIT-PL was successfully translated and adapted into Polish culture with satisfactory internal consistency (Cronbach's α = 0.83) and adequate test-retest reliability resulting in ICC2,1 = 0.89 (95% CI: 0.84-0.92). Measurement error was low (SEM = 0.76 and MDC = 2.10). Convergent validity of the CAIT-PL ranged from weak to strong using Spearman's correlation coefficient between CAIT-PL and FAOS subscales (r = 0.39-0.67; p < 0.05). Construct validity was confirmed.Conclusion: The Polish version of the Cumberland Ankle Instability Tool is a valid and reliable questionnaire for assessment of functional ankle instability among the Polish population.

The Cumberland Ankle Instability Tool is broadly used to assess functional ankle instability and its severity.The Polish version of Cumberland Ankle Instability Tool (CAIT-PL) showed adequate psychometric properties.The CAIT-PL is ready to be used in clinical practice and for research applications in Polish-speaking population.

Do Highly Trained Mountain Runners Differ from Recreational Active Non-Runners on Range of Motion and Strength in the Hip and Ankle as Well as Postural Control?

The rules governing mountain running force athletes to implement into their training programmes uphill and downhill running on unstable surfaces, which are demanding for hip and ankle as well as for the postural control system. The aim of the present cross-sectional study was to compare highly trained mountain runners (MR) and recreational active non-runners (NR) on range of motion (ROM) and strength in the hip and ankle, as well as dynamic postural control. Thirty MR and thirty-two NR were included in the study. ROM was assessed using a digital inclinometer. Strength was measured using a hand-held dynamometer. Postural control was evaluated using the lower quarter Y-balance test (YBT-LQ). The results showed that MR, in relation to NR, had statistically significant smaller hip external rotation ROM (p = 0.007), lower hip external rotator (p = 0.006) and extensor (p = 0.023) strength and greater normalised anterior reach in the YBT-LQ (p = 0.028). Mountain running training may reduce hip external rotation ROM as well as hip external rotator and extensor strength. Moreover, such training may improve postural control. MR should implement exercises targeted at developing hip ROM and strength. Furthermore, it seems that mountain running training may be a good way to improve postural control.

68 Ga-Prostate-Specific Membrane Antigen PET/CT in Ovarian Tumors : Potential to Differentiate Benign and Malignant Tumors Before Surgery: A Preliminary Report.

PURPOSE OF THE REPORT: Ovarian cancer is usually diagnosed in an advanced stage of disease due to the absence of specific symptoms and a lack of sensitive diagnostic methods. Prostate-specific membrane antigen (PSMA) is expressed on prostate cancer cells but can be found in other tumors such as ovarian cancer.The aim of this pilot study was to evaluate the feasibility of using 68 Ga-PSMA-11 PET/CT in detection of ovarian neoplasm before surgical treatment. PATIENTS AND METHODS: Eight women with mean age of 56.0 ± 16.2 years were included in the study. All patients underwent transvaginal ultrasound followed by CT scan of the chest and abdomen as qualification for surgery. Within a 1-week interval, PET/CT was performed on a Siemens Biograph scanner, 60 minutes after injection of 2 MBq/kg 68 Ga-PSMA-11. RESULTS: In 3 cases (37.5%), the 68 Ga-PSMA-11 PET/CT was positive, whereas histological examination confirmed 2 serous ovarian cancer cases and 1 ovarian borderline tumor. The SUV max in the serous ovarian cancer was 8.7 and 4.1, and in the borderline ovarian tumor, it was 13.8. No correlation was found between antigen CA-125 level and 68 Ga-PSMA expression. Range of tumor SUV max was not correlated with stage of disease. The remaining 62.5% (5/8) were negative in 68 Ga-PSMA-11 PET/CT, and histopathology confirmed benign pelvic tumor. CONCLUSIONS: The initial experience supports the potential to use 68 Ga-PSMA-11 in ovarian cancer to differentiate malignant and benign tumors before surgery.This study was approved by the Ethical Committee of the Medical University of Warsaw (KB/2/A/2018).

Chemotherapy of HER2- and MDM2-Enriched Breast Cancer Subtypes Induces Homologous Recombination DNA Repair and Chemoresistance.

Analyzing the TCGA breast cancer database, we discovered that patients with the HER2 cancer subtype and overexpression of MDM2 exhibited decreased post-treatment survival. Inhibition of MDM2 expression in the SKBR3 cell line (HER2 subtype) diminished the survival of cancer cells treated with doxorubicin, etoposide, and camptothecin. Moreover, we demonstrated that inhibition of MDM2 expression diminished DNA repair by homologous recombination (HR) and sensitized SKBR3 cells to a PARP inhibitor, olaparib. In H1299 (TP53-/-) cells treated with neocarzinostatin (NCS), overexpression of MDM2 WT or E3-dead MDM2 C478S variant stimulated the NCS-dependent phosphorylation of ATM, NBN, and BRCA1, proteins involved in HR DNA repair. However, overexpression of chaperone-dead MDM2 K454A variant diminished phosphorylation of these proteins as well as the HR DNA repair. Moreover, we demonstrated that, upon NCS treatment, MDM2 K454A interacted with NBN more efficiently than MDM2 WT and that MDM2 WT was degraded more efficiently than MDM2 K454A. Using a proliferation assay, we showed that overexpression of MDM2 WT, but not MDM2 K454A, led to acquisition of resistance to NCS. The presented results indicate that, following chemotherapy, MDM2 WT was released from MDM2-NBN complex and efficiently degraded, hence allowing extensive HR DNA repair leading to the acquisition of chemoresistance by cancer cells.

Zebrafish Microenvironment Elevates EMT and CSC-Like Phenotype of Engrafted Prostate Cancer Cells.

To visually and genetically trace single-cell dynamics of human prostate cancer (PCa) cells at the early stage of metastasis, a zebrafish (ZF) xenograft model was employed. The phenotypes of intravenously transplanted fluorescent cells were monitored by high-resolution, single-cell intravital confocal and light-sheet imaging. Engrafted osteotropic, androgen independent PCa cells were extravasated from caudle vein, invaded the neighboring tissue, proliferated and formed experimental metastases around caudal hematopoietic tissue (CHT) in four days. Gene expression comparison between cells in culture and in CHT revealed that engrafted PCa cells responded to the ZF microenvironment by elevating expression of epithelial-mesenchymal transition (EMT) and stemness markers. Next, metastatic potentials of ALDHhi cancer stem-like cells (CSCs) and ALDHlow non-CSCs were analyzed in ZF. Engraftment of CSCs induced faster metastatic onset, however after six days both cell subpopulations equally responded to the ZF microenvironment, resulting in the same increase of stemness genes expression including Nanog, Oct-4 and Cripto. Knockdown of Cripto significantly reduced the vimentin/E-cadherin ratio in engrafted cells, indicating that Cripto is required for transduction of the microenvironment signals from the ZF niche to increase mesenchymal potential of cells. Targeting of either Cripto or EMT transcriptional factors Snail 1 and Zeb1 significantly suppressed metastatic growth. These data indicated that zebrafish microenvironment governed the CSC/EMT plasticity of human PCa cells promoting metastasis initiation.

Electrical hippocampal activity during danger and safety signals in classical conditioning in the rat.

The effect of stimuli predicting danger (DS) and safety (SS) in Pavlovian aversive conditioning on hippocampal local field potentials (LFP) was studied in 25 partially restrained adult male rats (Long-Evans). DS lasting 5 s preceded tail-shock, while SS overlapping DS during DS last 3 s predicted omission of shock. The power spectra of LFPs during trials were analyzed in theta and delta frequency bands. In DS, theta frequency during the last 3 s was lower that in first 2 s. In danger and safety situation theta peak frequency was different for dorsal CA1 activity (5.99 Hz vs. 6.86 Hz, respectively) while delta peak frequency was different for ventral CA1 (1.56 Hz vs. 1.07 Hz) for the last 3 s of trial. Differences in theta frequency in danger and safety situation may reflect differences in sensory processing during induced emotional states and/or related differences in motor behavior.

Hydrothermal carbonization of biogas digestate: Effect of digestate origin and process conditions.

Hydrothermal carbonization (HTC) solids were produced in a 250-ml batch reactor from three different digestates at varying temperatures (170, 190, 210, 230, and 250 °C) and 2 and 5 h reaction time. Three potential feedstocks of biogas plants were tested: organic household waste, cow manure, and energy crops. The proximate composition, elemental composition, heating values, and thermal stability of the HTC solids were characterized. The dry ash-free basis yields and carbon recovery of HTC solids decreased with increasing HTC temperature and time. The HTC solids from energy crop digestate had the highest yields (dry ash-free basis) except at the temperature of 250 °C. Increased HTC severity was found to have a positive effect on the dry basis carbon content and leads to a higher heating values (HHV) of cow manure digestate HTC solids. The preferred reaction condition for organic household waste digestate and energy crop digestate HTC solids was found to be 210 °C for 5 h, because of the highest HHV (dry basis) and the overall combustion performance was reached. Enhancing HTC temperature increased the peak intensity of functional groups of cow manure digestate HTC solids. Scanning electron microscopy imaging showed the fibrous structure of the plants from the digestates was mostly deconstructed at 250 °C-5 h. Experimental results indicated that both digestate origin and HTC condition influenced the properties of the HTC solids.

Diverse and cancer type­specific roles of the p53 R248Q gain­of­function mutation in cancer migration and invasiveness.

Gain­of­function (GOF) mutations in the TP53 gene lead to acquisition of new functions by the mutated tumor suppressor p53 protein. A number of the over­represented 'hot spot' mutations, including the ones in codons 175, 248 or 273, convey GOF phenotypes. Such phenotypes may include resistance to chemotherapeutics or changes in motility and invasiveness. Whereas the prevalent notion is that the acquisition of the p53 GOF phenotype translates into poorer prognosis for the patient, the analysis of a human somatic p53 mutations dataset demonstrated earlier tumor onset, but decreased frequency and altered location of metastases in patients with the p53­R248Q allele. Therefore, the GOF activities of p53­R248Q and p53­D281G were analyzed in triple negative breast cancer MDA­MB­231 and lung adenocarcinoma H1299 cell lines with regard to invasive and metastatic traits. The expression of p53­D281G increased the motility and invasiveness of the lung cancer cells, but not those of the breast cancer cells. In contrast, the expression of p53­R248Q decreased the motility and invasiveness of the breast and lung cancer cells in a p53 transactivation­dependent manner. The intravenous xenotransplantation of MDA­MB­231 cells expressing p53­R248Q into zebrafish embryos resulted in an alteration of the distribution of cancer cells in the body of the fish. In p53­R248Q­expressing H1299 cells a decrease in the expression of TCF8/ZEB1 and N­cadherin was observed, suggesting partial mesenchymal­to­epithelial transition. In the two cell lines expressing p53­R248Q a decrease was noted in the expression of myosin light chain 2, a protein involved in actomyosin­based motility. To the best of our knowledge, the present study is one of only few reports demonstrating the mutated p53 GOF activity resulting in a decrease of a malignant trait in human cancer.