The p62 protein, also called sequestosome 1 (SQSTM1), is a ubiquitin-binding scaffold protein. The expression of p62 was statistically higher in carcinoma compared to non-neoplastic mammary glands. Our results, although preliminary, are similar to the results of breast cancer, therefore, also in the cat, p62 could be considered a possible oncotarget.The p62 protein, also called sequestosome 1 (SQSTM1), is a ubiquitin-binding scaffold protein. In human oncology, although the interest in the function of this protein is recent, the knowledge is now numerous, but its role in tumorigenesis is not yet clear. This preliminary study aims to evaluate the immunohistochemical expression of p62 in 38 cases of feline mammary carcinoma with different grades of differentiation and in 12 non-neoplastic mammary gland tissues, to assess the expression level and a possible correlation with malignancy. The expression of p62 was statistically higher in carcinoma compared to non-neoplastic mammary glands: 28 feline mammary carcinomas (73.7%) had a high p62 expression score, three (7.9%) had a moderate expression, while seven cases (18.4%) had a low expression. The grade of the differentiation of the carcinoma was not correlated with the p62 expression. This study represents the first approach in feline oncology that correlates p62 expression in feline mammary carcinoma. Our results, although preliminary, are similar to the results of human breast cancer, therefore, also in the cat, p62 could be considered a possible oncotarget. The p62 protein, also called sequestosome 1 (SQSTM1), is a “scaffold” protein (capable of bringing other proteins together), fundamental in the process of autophagy, linked to ubiquitin, which polymerizes through an N-terminal PB1 domain and which can interact with ubiquitinated proteins through the C-terminal UBA domain ,4.In human oncology, the cellular mechanism of autophagy has only recently been studied to understand its possible involvement in the carcinogenetic process. Autophagy is a non-selective mechanism, responsible for the degradation of the majority of long-lived cellular proteins and cell organelles . VariousThe metabolism of cancer cells is very different from that of normal cells: the high rate of cell proliferation, the increase in cell size, and the simpler enzymatic mechanisms require much oxygen and nutrients with the consequent production of high quantities of toxic compounds. Cancer cells are therefore able to reprogram their metabolism and activate the autophagy process in the various stages of the tumor growth .Although the interest regarding the function of p62 is quite recent, in human oncology, several studies have been published. Nevertheless, the role of p62 in tumorigenesis is not very clear. p62 is considered a protein whose function arises at the crossroads between autophagy and apoptosis ,11,12, aIn veterinary medicine, knowledge of the role of p62 in tumors is still limited, therefore, the purpose of this study is to evaluate the immunohistochemical expression of p62 in feline mammary carcinoma with different degrees of differentiation and in non-neoplastic mammary glands to highlight how this protein is expressed and if there is a correlation with the degree of the malignancy of the tumor. Mammary neoplasia in cats is less common compared to dogs, but when it occurs, most of the cases are malignant and histologic grading has a prognostic value .This study included fifty cases: 12 post-mortem non-neoplastic mammary glands and 38 mammary carcinomas, of which 34 of the simple type and four of the ductal type were collected from 2012 to 2021 at the Pathology Unit of the School of Biosciences and Veterinary Medicine of the University of Camerino . From the same archive came reports and anamnesis of the cases. Ages ranged from 4 to 16 years, with a median age of 11.4 years, and all were European shorthair females. The specimens were fixed in 10% neutral buffered formalin, subsequently processed by an automatic histoprocessor, and embedded in paraffin. Serial sections (3 µm) were stained with hematoxylin and eosin (HE).2. To each tumor, a score from 0 to 3 was assigned, and thus they were subsequently classified as grade I or low grade (score of 0), grade II, or intermediate cancers (score of 1), and grade III or higher cancers (score of 2–3).All feline mammary carcinoma cases included in the study underwent a histological grading system, following the more recent system proposed by Mills et al.  based onImmunohistochemical analysis was carried out with a standard ABC-peroxidase method on 3 µm sections, as described by Di Cerbo et al. . The lab2 was analyzed for the semiquantitative evaluation. A score was then assigned to each sample according to positive cell percentage and their signal intensity. The scoring system used was that described by Mariotti et al., 2019. [As a secondary antibody, a goat anti-rabbit biotinylated antibody was used along with DAB (diaminobenzidine) as the chromogen. Normal mouse pancreas was used as a positive control , while r., 2019. Positive cell percentage was based on a 5-point scoring system: 0 = no positive cell, 1 = ≤20%, 2 = between 21% and 50%, 3 = between 51% and 70%, 4 = ≥71%. For each microscopic field, a count of immunopositive and negative cells was carried out and then transformed in percentage. The intensity of immunostaining was classified to a 4-point scoring system: 0 = no staining, 1 = low-intensity staining, 2 = moderate-intensity staining, 3 = high-intensity staining. In those samples where the intensity was heterogeneous, the score chosen was the predominant one in the sample itself. The overall score assigned to each case is derived by the multiplication of cell positivity and intensity signal score, with a minimum score of 0 and a maximum score of 12. The equivocal cases were evaluated by two authors (G.E.M. and F.M.) to establish the score. Scores between 1 and 2 were considered as a low expression of p62, those from 3 to 4 as intermediate, and those greater than 5 as high expression.p < 0.05.Data were analyzed using GraphPad Prism 8 software . All data are presented as the means ± standard deviation (SD). Differences between the score of non-neoplastic patients and those with mammary carcinoma and differences between feline mammary carcinoma cases with lymphovascular invasion were analyzed using a Mann–Whitney test, while differences among different histological grades were analyzed using a Kruskal–Wallis’s test followed by Dunn’s multiple comparison test. The level of significance was set at * Eight cases of non-neoplastic feline mammary gland tissue studied were negative for p62. Widespread negativity was observed in the epithelial cells of the mammary gland A; four cThe histological grading of the 38 cases of feline mammary carcinomas studied was as follows: five cases were low-grade GI (13.2%), 10 intermediate-grade GII (26.3%), and 23 high-grade GIII (60.5%).Twenty-eight cases (73.7%) had a high p62 expression score (≥5), three (7.9%) had a score between 3 and 4, while seven cases (18.4%) had a low score between 1 and 2 . PositivThe correlation between the p62 expression score and the histological grade of the mammary carcinomas is reported in This preliminary study aimed to evaluate the immunohistochemical expression of the p62 protein in feline mammary carcinoma and feline mammary non-neoplastic tissue, taking into consideration tumors of different histological grades with different prognoses.In this study, we observed a high positivity of feline mammary carcinoma for p62. These preliminary observations could suggest a correlation between the expression of p62 and the carcinogenic process since in carcinomas there is a high expression of the protein as opposed to non-neoplastic mammary gland tissue. Although is not possible to draw a definitive conclusion about the expression of p62 in feline mammary carcinomas, the statistical difference observed between mammary carcinoma and non-neoplastic tissue is a result to be considered.In this study, no correlation was found between the expression of p62 and the different grades of mammary carcinoma, but the data obtained seem to show a certain tendency towards the loss of p62 from grade I carcinoma to grade III. We also wanted to evaluate the expression in tumors presenting lymphovascular invasion, a histological feature to be considered a statistically significant prognostic factor , and we In general, although the data are preliminary, they seem to agree, in terms of p62 expression levels, with those of female breast cancer. In fact, two studies demonstrated a higher expression of p62 in carcinomas compared to normal breast tissue ,26.Few studies on p62 and cancer have been performed in veterinary medicine. A study on canine cutaneous mast cell tumors demonstrated nuclear immunoreactivity in low-grade tumors and cytoplasmic immunoreactivity in high-grade tumors . In our A preliminary study on simple canine mammary carcinoma was more recently published and a reduction in p62 reactivity was observed as the degree of malignancy increased  .To date, both in human medicine and in veterinary medicine, the knowledge of the role of p62 in oncogenesis is still insufficient and not exactly delineated, partly due to the multiple activities of this protein, but from what we know so far, we can consider this protein as an independent tumor marker and not an oncogene .However, it has been shown that after oncogenic transformation, p62 is necessary for tumor initiation and tumor progression: the induced formation of Her2- breast cancer and the RAS-induced formation of lung cancer are hindered in p62 knockout animals ,30. FurtAnother very interesting study area, in which p62 could be involved, is the relationship established between neoplastic epithelial cells, the inflammatory response, and activated fibroblasts associated with epithelial cancer. Studies on human prostate cancer and breast cancer have suggested important metabolic differences between neoplastic cells and fibroblasts, as neoplastic cells are anabolic with a high expression of p62 while stromal cells are catabolic and lose p62 ,36. AccoThis study confirms the importance of the use of the cat as a translational spontaneous tumor model, especially for feline mammary carcinoma, whose analogies with human breast cancer are wildly acknowledged . This pr


In the cellular response to stresses, the tumor suppressor p53 is activated to maintain genomic integrity and fidelity. As a transcription factor, p53 exhibits rich dynamics to allow for discrimination of the type and intensity of stresses and to direct the selective activation of target genes involved in different processes including cell cycle arrest and apoptosis. In this review, we focused on how stresses are encoded into p53 dynamics and how the dynamics are decoded into cellular outcomes. Theoretical modeling may provide a global view of signaling in the p53 network by coupling the encoding and decoding processes. We discussed the significance of modeling in revealing the mechanisms of the transition between p53 dynamic modes. Moreover, we shed light on the crosstalk between the p53 network and other signaling networks. This review may advance the understanding of operating principles of the p53 signaling network comprehensively and provide insights into p53 dynamics-based cancer therapy. Cells exhibit powerful information processing capabilities when they are exposed to various stresses such as DNA damage, hypoxia, and nutrition deficiencies ,2. In-deTranscription factors (TFs) are proteins that can bind to the promoters of target genes specifically to regulate their expression. Due to their versatility, TFs are usually considered as information processing centers. As one of the most important tumor suppressors, p53 mainly acts as a transcription factor to maintain genomic integrity by inducing a large number of target genes in cellular response to stresses . DiffereA fundamental strategy in understanding the underlying mechanisms of various dynamics is to seek out the corresponding regulatory networks in response to specific stresses. Based on the given functions of the network motifs, we can make some speculations about which kind of motif is responsible for the observed dynamics. The alternate activation of p53 and Mdm2 is responsible for p53 oscillations upon ionizing radiation (IR) . DiffereIn this review, we discuss recent advances in understanding the underlying mechanisms in the generation of p53 dynamics and the decision of cell fates in response to stresses. We summarize the latest findings in two aspects: encoding—how DNA damage is encoded into p53 dynamics and decoding—how cell fate is decoded from p53 dynamics. To give an overview of p53 response to DNA damage, we summarize the p53 signaling network including the damage types, sensors, upstream circuits, dynamics, transcription, and downstream circuits associated with different cell fates . MoreoveThe integrity of the genome is challenged by genotoxic stresses. Failure in cellular response to such stresses is closely related to carcinogenesis. During the cell cycle, cells are subject to transient and intrinsic DNA damage during mitosis. Extrinsic DNA damage agents mainly include antitumor drugs , etoposide, cisplatin, and NCS (Neocarzinostatin)) and irradiations. These damage agents can produce single- or double-strand breaks (SSBs or DSBs) in DNA. As the input to the p53 signaling network, different types of DNA damage activates the downstream pathways specifically, leading to different cellular outcomes .SSBs are breaks in one strand of the DNA double helix, mainly caused by UV radiation. They can be resolved by homologous recombination (HR) and alternative homologue-mediated SSB repair pathways . DSBs arCells sense DNA damage by activating several kinases including ATM, ATR, and DNA-PKcs, which are accountable for detecting damage lesions via the MRN (MRE11-RAD50-NBS1) complex and transmitting damage signals to the tumor suppressor p53. Upon DNA damage, ATM is specially activated by DSBs through phosphorylation, regulating DSBs repair and activating p53 . Based oATM, ATR, and DNA-PKcs are all members of phosphatidylinositol 3-kinase-related kinases (PIKKs) family. Although they can sense DNA damage independently, there is a crosstalk between them in some cases . DNA-PKcUpon DNA damage, p53 levels vary over time and exhibit rich dynamics . Lahav eIn response to UV-induced DNA damage, a single extended pulse of p53 is induced and its width and height increase with the UV dose . As a reNotably, the above results were observed in MCF7 cells, which were later found to be insensitive to a spectrum of antitumor drugs including etoposide, Nutlin-3a, and 5-Fluorouracil . Indeed,There exists another mode of two-phase dynamics of p53: cells exhibit a high-amplitude p53 pulse in the first phase and undergo low-amplitude p53 oscillations in the second phase . This kiDifferent p53 dynamics in various cell lines have inspired the investigation into whether such differences in p53 dynamics exist in different tissues or species. Lahav and coworkers observed that the radiosensitivity corresponding to p53 dynamics shows remarkable differences between tissues. The small and large intestines are relatively insensitive to p53 levels peaking at 2–3 h followed by decreasing in its levels, while lymphoid organs, including the spleen and pancreas, are more sensitive to radiation with sustained p53 expression . In anotThe mathematical modeling on p53 network provides a theoretical perspective to understand the generation mechanism of p53 dynamics. The negative feedback loop between p53 and Mdm2 governs p53 oscillations upon DNA damage . NeverthThe core status of the p53-Mdm2 loop in p53 regulation is challenged by the connection of p53 pulses to the pulses of the damage sensor ATM . p53-indLong-term measurements revealed that p53 oscillations exhibited marked heterogeneity, especially in the oscillation amplitudes . Based oThe generation of basal dynamics of p53 depends heavily on an excitable mechanism. A hallmark of an excitable system is that a transient input is sufficient for triggering a full response. Thus, similar to the case of cellular response to extrinsic damage, a complete p53 pulse will be initiated by transient damage during mitosis . BatchelThe coupling of two negative feedback loops is important for producing some specific properties of the p53 system, such as excitability . It has mdm2. This positive feedback loop may control the amplitude of p53 pulses [Interconnected positive and negative feedback loops (IPNFLs) are another prevalent coupling pattern in producing p53 pulses  Figure . The two3 pulses .In response to DNA damage, p53 is gradually activated to perform different functions and different feedback loops modulate p53 dynamics in this process. In modeling, it was assumed that there exist different forms of p53 based on its phosphorylation status ,16,19,47Moreover, other functional factors also play some roles in the encoding of p53 dynamics. Multiple miRNAs, including miR-605, miR-192, miR-29a, and miR-34a can directly or indirectly repress Mdm2, thus forming a p53-miRNA-Mdm2 positive feedback loop ,55. ThesIn response to stresses, p53 induces hundreds of target genes to modulate a wide range of cellular processes including cell cycle arrest, apoptosis, senescence, DNA repair, and metabolism ,60. ThisOver the past decade, much effort has been put into exploring the selective expression of the target genes by oscillatory p53. By experimental measurements, the amplitudes of p53 pulses were found to delineate promoter activation thresholds, while pulse frequencies were differentially filtered by the promoters of the target genes . These rUnder p53 oscillations, p21 mRNA tightly follows p53 pulses, while p21 protein has three different patterns: oscillation, slow, and rapid accumulation . The difAdditional mechanisms, including different activation thresholds and the presence of feedforward loops, should contribute to modulating the exclusive induction of targets under specific p53 dynamics . MeanwhiIn addition to the dynamic patterns of p53, there are multiple levels of regulation that can help p53 select targets to execute appropriate cellular responses . Upon DNThe acetylation status assists p53 in preferentially activating a specific cell fate, such as apoptosis, highlighting the importance of post-translational modifications in regulating the target selectivity of p53 . Gu et arylation . These mrylation ,76.The selective induction of p53 targets between cell cycle arrest, senescence, and apoptosis has been extensively explored. p53 leads to cell cycle arrest by inducing p21 ,78. The A great deal of modeling work has focused on the choice between survival and apoptosis in cellular response to stresses. In general, differential activation of BAX, PUMA, Apaf1, Casp3, and p21 can be exploited to indicate different cellular outcomes in the models ,82. BaseThe regulation of apoptosis becomes complex when multiple factors are involved. Activated E2F-1 induces ASPP, which promotes the expression of BAX to trigger apoptosis . p53-ind pathway . A Boole pathway . Similarp53 shows persistent pulses at low etoposide doses but exhibits monotonic increasing at rather high doses in U-2 OS cells . It is iIt is also important to investigate how p53 oscillations terminate after DNA repair. Recently, it has been proposed that p53 undergoes an intermediate state between early high-frequency oscillations and late low-frequency pulses and finally returns to a basal state  Figure c. It canIt was recently reported that the transition dynamics of p53 from oscillations to high rising in etoposide-treated U-2 OS and A549 cells may sequentially undergo damped oscillations and a rising with a moderate platform  Figure f. The trThe study of transition dynamics suggests that we can look for clues to kill cancer cells by amplifying p53 signaling. We can choose an agent that is sensitive to the specific cells. For example, MCF7 cells are sensitive to UV light but not to NCS and Etoposide . In factThe p53 network has a key role in response to cellular stresses, creating scenarios in which the p53 network couples with other signaling networks. The establishment of a coupling relationship is supported by several aspects: (I) coactivation by the same stimulus ; (II) sharing some key regulatory factors ; and (III) coregulation of the same cellular process. The coactivation of multiple signaling networks is the basis of network coupling, and the abundance of coregulators determines the degree of activation of individual networks, thus affecting the dynamic properties and functions of each network .Hypoxia can activate both p53 and HIF-1, thus causing an interplay between these two transcription factors. However, their activation is different and determined by the severity of hypoxia: HIF-1 is activated to promote glycolysis and angiogenesis upon mild and moderate hypoxia, while only upon severe hypoxia or anoxia, p53 is induced to trigger apoptosis . Our mod factors ,96,97. TIt was reported that the interference of NF-pathways . The inh and p53 . Furthernd NF-κB . Indeed,p300/CBP . An expln of p53 . It remaE2F-1 is a vital transcription factor that primarily regulates the cell cycle progression from G1 to S phase. In early G1, its transcriptional activity is repressed by pRB, while in late G1, E2F-1 is released and activated as pRB is phosphorylated by the Cyclin/CDKs (cyclin-dependent kinase) complex, driving the cells from G1 phase to S phase . Due to It is a challenge to understand the coactivation of p53 and TGF-activity ; at the activity . These trylation ,108. TGF and Zeb . The mut and Zeb . Strikinl of EMT . It is all cycle . TherefoOver the last 20 years, the development in experimental techniques has advanced the measurement of p53 dynamics in individual cells. In general, p53 dynamics depend on the type of stimulus, cell, tissue, and species. In addition, the detection methods have also been found to impact the dynamics of p53 obtained by experiments . Both ex


Gliomas are extremely debilitating malignant brain tumors with very limited response to therapies. The initiation and progression of gliomas can be attributed to several molecular abnormalities, such as mutations in important regulatory networks. In this regard, the mitogen-activated protein kinases (MAPKs) arise as key signaling pathways involved in cell proliferation, survival, and differentiation. MAPK pathway has been altered in most glial tumors. In glioma cells, the activation of p38 MAPK contributes to tumor invasion and metastasis and is positively correlated with tumor grade, being considered a potential oncogenic factor contributing to brain tumorigenesis and chemotherapy resistance. Hence, a better understanding of glioma pathogenesis is essential to the advancement of therapies that provide extended life expectancy for glioma patients. This review aims to explore the role of the p38 MAPK pathway in the genesis and progression of malignant brain tumors. Malignant brain tumors are rapidly growing cancer with high invasion potential to surrounding healthy brain structures, which include gliomas and neuronal or mixed neuronal-glial tumors . GliomasThe initiation and progression of gliomas can be attributed to several molecular abnormalities, such as mutations in important regulatory networks . The mitAmong the MAPK pathways, p38 mitogen-activated protein kinase (p38 MAPK) signaling was identified as a mediator of stress and inflammation responses . This paConsidering the importance of the signaling pathways involved in the genetic and molecular alterations in the onset of cancer, describing the role of the p38 MAPK pathway in tumor microenvironment and in the progression of gliomas becomes a key strategy to comprehend and propose new treatments for malignant brain tumors.Malignant brain tumors are aggressive and impact directly patients’ life quality . GliomasGlioblastoma (GBM) is the highest grade of diffuse astrocytoma and is considered incurable . DespiteIt is well known that molecular characteristics inside the tumor microenvironment (TME) indicate the sensitivity of tumors to therapy. Initially, tumorigenesis is related to the gain of numerous genetic mutations, which provide malignant cells resistant to the growth-inhibitory and apoptotic signals . All GBMIn general, tumors are complex tissues composed of distinct cell types that interact with each other forming an intense signaling network . The gliHypoxia is considered a hallmark of GBM, and a critical factor associated with mutations in multiple signaling pathways that affect the tumor landscape . Solid tProtein kinases are enzymes that have the property to catalyze protein phosphorylation through the transfer of a phosphoryl group from ATP and GTP to threonine, serine , tyrosine , both serine/threonine and tyrosine  . The phoα, β, γ, and δ), and ERK5. Atypical MAPKs have nonconforming particularities and consist of ERK3/4, ERK7, and Nemo-like kinase (NLK)  .The activation of MAPKs and their downstream targets plays a pivotal role in different signaling cascades involved in transcription, development, differentiation, migration, cell death, and many other critical cellular functions . DetrimeMAPKs signaling cascades present recurrent contributions to oncogenesis, tumor progression, and drug resistance, which suggests that its pharmacological modulation can be a promising strategy in the development of cancer therapies . It is wDiscovered on a pharmacological screen in 1994, p38 MAPK is a signal transduction pathway that plays a key role in cellular adaptation to extracellular stimuli . Its actAfter extracellular signals, activation of the p38 pathway usually proceeds through a classical phosphorylation cascade, where a MAPKKK  phosphorylates and activates the MAPKKs specifics of p38 MAPK, MKK3, and 6, which in turn, mediate the activation of the different p38 MAPK isoforms . The p38The p38 MAPK family compromises four isoforms: p38α , p38β , p38γ , and p38δ  . The fouIn recent years, the involvement of p38 MAPK in cancer has been widely described . This paSome studies have suggested a role for p38 MAPK in mediating pathways that lead to cell apoptosis and growth inhibitory signals, particularly in promoting cell cycle arrest and differentiation, which supports the idea that the stress-activated kinase is a tumor suppressor . HoweverMAPKs are expressed in various types of malignant brain tumors . Besidesin vitro and in vivo. Irradiation is often part of the GBM treatment scheme, although, in cells that present mutant PTEN it was described that irradiation can activate p38/Akt and PI3K/Akt signaling pathways, increasing MMP-2 expression and intensifying invasiveness  which activates the p38 MAPK pathway (Long noncoding RNAs (lncRNAs) are transcripts with more than 200 nucleotides that commonly do not encode proteins, many of them have been characterized as oncogenes or tumor suppressors in cancer . The MRC pathway .Nowadays it is known that, in tumor progression, gliomas orchestrate the immune system to a protumoral phenotype. The p38 MAPK is also correlated with immune and inflammation signaling, being responsible for the production of cytokines in the TME . Wang etin vitro  release . Other sin vitro . Figure Several small-molecule protein kinase inhibitors have been tested for cancer treatment, demonstrating interesting effects on the immune system. Among these, there is the EGFR tyrosine kinase inhibitor gefitinib, and other inhibitors focusing on multiple targets, like imatinib, sorafenib, and sunitinib . Regardlvia Nrf2 signaling inhibition, indicating that p38 MAPK/Nrf2 activation is a network involved in temozolomide glioma cells resistance  was used in combination with temozolomide, it reduced temozolomide antitumoral effects . BesidesSeveral clinical trials targeting p38 MAPK in cancer are currently underway. Ralimetinib (LY2228820), for example, is a potent and selective inhibitor of p38α and p38β and has been tested as a single agent and in combination with chemotherapeutic agents for the treatment of ovarian cancer, GBM, and metastatic breast cancer . An impoBecause of the wide-ranging controlling role of p38 in different cellular processes, the possibility of side effects resulting from an undesired pharmacological activity is a relevant concern for the p38 inhibitors. There are many discontinued p38 inhibitors that failed due to safety concerns, presenting adverse effects directed mostly to the hepatic system and skin . Other iMalignant brain tumors are debilitating diseases with a dismal prognosis and extremely limited response to therapies. The aggressiveness of GBM has been marked by several signaling pathway dysregulations that are implicated in cell proliferation, survival, migration, and angiogenesis. Tumor cells are heterogeneous, so it is also important to note that targeting a mediator involved in multiple pathways would constitute a more efficient therapy. MAPKs are shown to support numerous processes of cell survival. The p38 MAPK pathway is a key route capable to influence different frameworks of cellular survival mechanisms such as inflammation, proliferation, migration, invasion, and ROS signaling. Briefly, the p38 MAPK is likely to be an oncogenic factor contributing to GBM initiation, progression, and chemotherapy resistance. Hence, a better understanding of the GBM tumor microenvironment is essential to the advancement of therapies that provide extended life expectancy for patients. Thus, targeting key components of these signaling pathways using small-molecule inhibitors, such as p38 inhibitors and antibodies, could provide progress in the treatment of gliomas.


The p53 protein is a tumor suppressor encoded by the TP53 gene and consists of 393 amino acids with four main functional domains. This protein responds to various cellular stresses to regulate the expression of target genes, thereby causing DNA repair, cell cycle arrest, apoptosis, metabolic changes, and aging. Mutations in the TP53 gene and the functions of the wild-type p53 protein (wtp53) have been linked to various human cancers. Eight TP53 gene mutations are located in codons, constituting 28% of all p53 mutations. The p53 can be used as a biomarker for tumor progression and an excellent target for designing cancer treatment strategies. In wild-type p53-carrying cancers, abnormal signaling of the p53 pathway usually occurs due to other unusual settings, such as high MDM2 expression. These differences between cancer cell p53 and normal cells have made p53 one of the most important targets for cancer treatment. In this review, we have dealt with various issues, such as the relative contribution of wild-type p53 loss of function, including transactivation-dependent and transactivation-independent activities in oncogenic processes and their role in cancer development. We also discuss the role of p53 in the process of ferroptosis and its targeting in cancer treatment. Finally, we focus on p53-related drug delivery systems and investigate the challenges and solutions. The TP53 gene encodes a tumor suppressor protein called P53. The p53 protein was first identified in 1979 as a cellular protein that binds to the large T antigen of the simian virus (SV40) and accumulates in the nucleus of cancer cells. This protein contains domains of transcriptional activation, DNA binding, and oligomerization. The encoded P53 protein responds to various cellular stresses to regulate the expression of target genes, thereby causing DNA repair, cell cycle arrest, apoptosis, metabolic changes, and aging. Mutations in this gene and the functions of the wild-type p53 protein (wtp53) have been linked to various human cancers .According to studies, Eight TP53 gene mutations are located in codons that make up 28% of all p53 mutations. Mutations in these alleles appear in many human cancers and tissues. These alleles are called hotspots . MutatioThe p53 protein is vital in maintaining DNA stability and preventing cancer. Normally, wild-type p53 protein (wtp53) binds to a negative regulator and is inactivated or degraded. However, when DNA damage occurs in a cell, the p53 protein is induced, causing the cell cycle to stop. This gives the cells a chance to repair, but if the damage is severe, the cells will develop apoptosis. The p53 (mtp53) gene mutation has been linked to various cancers Many mutations occur in the DNA binding domain of the p53 gene. On the other hand, the successful interaction of wtp53 protein with transcription factors facilitates the repression or activation of appropriate target genes. This process is disrupted by the presence of the defective mtp53 protein. Thus, converting mtp53 protein to wild-type p53 protein could be a promising way to prevent or reverse tumor progression .Among these, we can mention some small molecular compounds, such as RITA , which target p53. This molecular compound directly binds to p53, disrupts the interaction between p53 and MDM2, and prevents p53 degradation. RITA increases tumor suppressor expression and suppresses oncogene expression through p53 reactivation; As a result, it causes cell death mediated by p53 .The p53 is a biomarker for tumor progression and a perfect target for designing cancer treatment strategies. In wild-type p53-carrying cancers, abnormal signaling of the p53 pathway usually occurs due to other unusual settings, such as high MDM2 expression. These differences between cancer cell p53 and normal cells have made p53 one of the most important targets for cancer treatment .In recent years, studies in this field have shown the structure, function, and role of p53 in tumor formation and development. These basilar studies strongly support the design and development of new approaches to targeting the mutant and wild-type p53 in cancer treatment. For example, delivery of wild-type p53 by adenovirus infection kills cancer cells and affects and suppresses tumor growth in preclinical and clinical trials . Also, min vitro and in vivo  are observed at the amino acid sites 175, 245, 248, 249, 273, and 282  Freed-P. MeanwhiAs mentioned, two hotspots and common cancer mutations in p53 protein are amino acid sites 175 and 273. R175H and R273H are more prone to aggregation than wild-type (WT) p53, and their pathological aggregation can lead to various cancers. A recent study investigated the dynamic and structural properties of the R175H and R273H mutants in the p53 core domain (p53C) using extensive all-atom molecular dynamics simulations. In this regard, the researchers found that in both R175H and R273H mutations, the β-sheet structure is well preserved; however, a larger hydrophobic surface and a higher loop flexibility than WT p53C were seen, thus predisposing proteins to accumulate in the cell. Also, an allosteric pathway has been identified through which the R273H mutation increases the flexibility of the N-terminal region around loop 2. These results indicate mechanical insights into the high accumulation tendencies of R175H and R273H mutants .In cancer biology, the vital point about p53 is that; the p53 mutant protein is found in 50% or more of 50% of human cancers. In addition to losing function, the mutant p53 can have a dominant-negative effect on the remaining wild-type p53 allele and subsequently inactivate it by losing heterozygosity (LOH). Also, some p53 mutations have additive functions, which will cause the tumor to grow. In cancers in which wild-type p53 is conserved, it is usually in regulatory genes that encode the up or down pathways of p53; changes are observed .Among p53 mutants, missense mutations not only cause the mutated p53 protein to lose its wild-type (LOF) function and gain dominant-negative activity but also increase the function of the mutated p53, leading to the tumor’s more aggressive behavior and drug resistance .In the remainder of this section, we will discuss the relative contribution of wild-type p53 loss of function, including transactivation-dependent and transactivation-independent activities in oncogenic processes and their role in cancer development.The TP53 gene acts as a tumor suppressor and cellular stress sensor. In stress-free cells, p53 is targeted for degradation by E3 ubiquitin ligase MDM2 and kept at low levels. Stress signals, such as DNA damage, oncogene expression, and hypoxia relieve P53 from MDM2 inhibition. Mutations in human cancers that occur mainly in the domain of DNA binding and specific sequences; disrupt p53 activation. A mutation in the TP53 gene disrupts the binding capacity of DNA by disrupting the structure of the p53 protein; it also increases oncogenic function. The TP53 gene also encodes a transcription factor that is an essential factor in cancer prevention. Inactivation of this gene leads to the most common mutation in sporadic human cancers .Studies have shown that p53-R248Q expression reduces the motility and invasiveness of lung and breast cancer cells in a p53 transactivation-dependent manner. Indeed, in the cell lines expressing p53-R248Q, myosin light chain two expression decreases. This chain is a protein involved in actomyosin-based motility. Also, the expression of R248Q in Matrigel’s absence significantly reduces the migration rate in a trans-dependent manner .The wild-type p53 regulates the expression of more than 2,500 genes as a transcription factor. The p53 helps respond to various stresses such as DNA damage, hypoxia, and oncogene activation. Also, depending on the severity and duration of stress, it leads to cell cycle arrest or apoptosis . Some ofThe wild-type p53 dysfunction can disrupt autophagy signaling. In this regard, the nuclear transactivation-dependent activation of autophagy by p53 is often impaired due to inactivation or deficiency of p53 in the cytoplasm. However, the tumor can stimulate pro-autophagic functions. The downregulation of the mTOR set translates this activity. The researchers found that p53 may inhibit autophagy using direct protein-protein interactions. Based on the findings of a study, gain-of-function (GOF) mutant p53 proteins inhibit the autophagic pathway and increase the proliferation of pancreatic and breast cancer cells. This reaction has been accompanied by stimulation of AMPK-mTOR genes and suppression of DRAM, Beclin-1, ATG12, and sestrin genes .Although most studies on p53-mediated tumor suppression mechanisms are based on the activation of p53 target genes, other aspects of this issue include the performance of p53 independent of the reaction, which has been reported recently. Induction of apoptosis in mitochondria can be considered the best function of p53 independent of transactivation. The p53 protein can bind members of the BCL-2 family of anti-apoptosis to replace them with members of the BCL-2 family of pro-apoptotic. It can also directly activate BAK and BAX, infiltrate the external mitochondrial matrix, and stimulate apoptosis  Ho et a.The p53 protein, in addition to mitochondria, has functions other than transactivation in other sites, such as the cytoplasm and nucleus , which cIn a study of mouse models of intestinal cancer based on the Wnt signaling pathway caused by an ApcMin/+ mutation or deletion of Csnk1a1 in the proximal intestine, cancer was suppressed in the presence of p53R172H. It can be said that this repression is based on a mutation in the DNA binding domain that is responsible for the trans-activation of the target gene and is related to the p53 −/− model. This is related to the inhibition of TCF4 chromatin binding in the Wnt pathway and, in a sense, the suppression of this pathway and tumor-independent p53 activation. Further tests are needed to prove this mutant’s neomorphic activity or presence in wild-type p53 .in vivo. p53R178E may provide a potential mechanism for apoptosis independent of the transactivation induced by p53R178E, and it can localize in the mitochondria  that can be regulated in various ways, including levels of transcription factors and changes in the activity of antioxidant enzymes. The wild-type p53 regulates ferroptosis through lipid peroxidation, amino acid metabolism, and the biosynthesis of glutathione and phospholipids . Also, tIt is important to note that p53 sometimes reduces cellular susceptibility to ferroptosis. Studies have shown that wild-type p53 protein activates p21 protein in a transcription-dependent manner and delays the onset of ferroptosis . ResearcThere are two main pathways in the induction of ferroptosis. The cysteine-glutamate transporter is the first pathway, which includes erastin, sulfasalazine, and glutamate. The second pathway in the induction of ferroptosis directly inhibits glutathione peroxidase (GPX) activity and includes RSL3 and DP17 . In the One of the roles of erastin is to influence the voltage-dependent anion channel (VDAC). This ion channel is located in the mitochondria’s outer membrane . VDAC coC− system ; This was while the p533KR mutant cell mortality was very high (>90%). However, if the expression of SLC7A11 is excessive in p533KR mutant cells, treatment with erastin will significantly reduce the cell death rate (20%)  .SLC7A11 transcription were lost. This mutant model, called p534KR98, significantly reduces cell death and tumor inhibitory function during treatment with erastin. These results suggest that activating p53 by erastin may play an essential role in tumor inhibition by inhibiting transcription of SLC7A11 and stimulating the ferroptosis process  that inhibit SLC7A11 display significant radiation sensitizing effects on tumor organoids and xenografts derived from patients with p53 deficiency or mutation. Accordingly, activation of p53 and RT-induced ferroptosis leads to better clinical outcomes for RT in cancer patients, and it is suggested that these FINs be used in combination with radiotherapy to treat p53 mutant cancers (Radiotherapy (RT) breaks down two strands of DNA and produces reactive oxygen species (ROS); this causes the cessation of the cell cycle, aging, and cell death . Since t cancers .The frequency of p53 mutations in most cancers is about 50%, but this frequency varies depending on the type of cancer. For example, p53 mutation has been reported in cancers of laryngeal (40.4%), esophageal (43.1%), head and neck (40.6%), ovarian (47.8%), and colorectal (43.2%). However, the frequency of p53 mutation in a small number of cancers is less than 20%. For example, endocrine (14.6%), hematopoietic (12.7%), and cervical (5.8%) tumors have the lowest frequency of p53 mutations . TherefoProtein accumulation, instability, improper folding, and defective transport generally lead to cellular functions and processes dysfunction. Many of these protein accumulation diseases are caused by the wrong folding of the protein, possibly due to genetic mutations and environmental stress conditions . Accordiin vitro and in vivo. These peptides are for the region prone to p53 accumulation; amino acids 252–258 are designed. For example, research has been conducted on treating different models of ovarian cancer in vivo and in vitro and the potential of these peptides in accumulating p53 mutants , which has an inhibitory effect on histone deacetylase (HDAC), can directly affect mutant p53 and lead to p53 phosphorylation in serine 15 and cessation of cell division in the G2/M stage and finally tumor cell death. It should be noted that its role in inhibiting ovarian cancer cell lines is well known .According to recent studies, zinc deficiency induces oxidative DNA damage and is involved in developing the mutated p53 . ResearcSome of these methods have limitations and may not be used to treat all cancers associated with p53 accumulation. For example, in the designer peptide method, if wild-type p53 is somewhat compacted or unfolded, wild-type p53 structures may also be targeted. If this occurs in normal cells in the body, it can lead to systemic toxicity. Also, in therapies based on small molecules, peptides are rapidly eliminated from the bloodstream due to their short half-lives, small molecular weight, and size; as a result, more optimization is needed . Hence, Another treatment method is the production of gold nanoparticles (AuNPs). These nanostructures provide a robust and non-toxic delivery system for gapmers in cancer cells and significantly reduce mutant p53 proteins. They overcome chemoresistance to gemcitabine. The formulation of AuNPs consists of a combination of a mixed polymer layer of polyethylene glycol (PEG) and PEI and a layer-by-layer assembly of bPEI through a sensitive linker. These nanoparticles can bind oligonucleotides through electrostatic interactions and release them as glutathione .in vitro. These findings suggest that combining synthetic mRNA NP delivery strategy with other cancer treatments can be very effective (The redox response for efficient delivery of p53-encoding synthetic messenger RNA (mRNA) is another nanoparticle (NP)-based engineered platform. Based on the findings, synthetic p53-mRNA nanoparticles significantly delay the growth of p53-null HCC and NSCLC cells by inducing apoptosis and cell cycle arrest. Also, p53 restoration significantly improves the sensitivity of these tumor cells to everolimus, a mammalian target of rapamycin (mTOR) inhibitor that is not expressed in HCC and advanced NSCLC. Furthermore, co-targeting the mTOR and p53 signaling pathways lead to distinct antitumor effects in several animal models of HCC and NSCLC ffective .In addition to the above, monoclonal antibodies can be potent tools against p53-specific mutations and maximize treatment potential with personalized or precision medicine approaches .Since the wild-type P53 protein has important and extensive roles and functions within the cell; as a result of mutations in this protein, which can occur due to various factors, causes various types of cancer. According to the findings, the p53 mutant protein is found in 50% or more of 50% of human cancers. Therefore, specialized therapeutic approaches to target P53 mutants or pathways associated with this protein have received much attention recently Although research in this area needs further investigation, it is hoped that it will lead to proper and effective treatments for various types of cancer and pave the way for future studies.


Antimicrobial stewardship (AMS) is a critical global intervention aimed at optimising antimicrobial use and decreasing antimicrobial resistance (AMR) with pharmacists playing a pivotal role within AMS teams. However, AMS is not comprehensively taught in pharmacy curricula and little is known about the relevance of pharmacists’ training to meet AMS needs in South Africa.This study aimed to explore the attitudes, knowledge and perceptions of clinical pharmacists towards AMS participation and training in South Africa.This study was conducted among clinically practicing pharmacists in public and private healthcare sectors in South Africa.A quantitative exploratory research design was selected for this study. The study was conducted using a self-administered structured survey. Categorical variables were analysed using simple descriptive statistics. Mann–Whitney and Kruskal–Wallis tests were applied to determine differences between variables.p = 0.005), sector of employment (p = 0.01), position of employment (p = 0.015) and presence of AMS programmes (p = 0.004). Pharmacists indicated that their Bachelor of Pharmacy undergraduate studies inadequately prepared them for their role in AMS (median 4.3).Pharmacists demonstrated good attitudes knowledge and perceptions towards AMS (median 4.3). There was statistical significant differences in AMS participation between pharmacists of different years of experience  and workshops and insufficiently incorporated in undergraduate programmes.This study confirms that undergraduate pharmacy programmes inadequately prepare pharmacists for their role in AMS. Antimicrobial resistance (AMR) poses a major threat to the health of populations worldwide, puts achievements in medical advancements at risk and increases morbidity, mortality and the global economic burden  was approached to distribute the survey to its members. The SASOCP is a society formed in response to the resurgent interest in clinical pharmacy with 85 paid members at the time the survey was distributed. A sample size of 55 was obtained, yielding a response rate of 64.7%. Out of a maximum of 55 responses, not all respondents answered and rated each variable from questions two to nine. Hence, the number of responses received for each variable is reflected by a change in A self-administered structured survey was designed specifically for this study, incorporating questions from previous instrument designs of similar studies  of the University of the Witwatersrand, ethics clearance number M210937. A study information sheet explained the study in detail and the benefits and risks of participating in the study. Completion of the questionnaire was anonymous and data were confidentially maintained.n = 40) of the study population and male participants represented 27.27% (n = 15). The majority of the participants were between the ages of 30 and 39 years 45.45% (n = 25), have an M. Pharm qualification 65.46% (n = 36) with more than 10 years’ experience 50.91% (n = 28). Pharmacists in the private sector represented 81.82% (n = 45) of the responses. The majority of participants were employed as pharmacists 32.73% (n = 18), followed by 29.09% (n = 16) clinical pharmacists and 27.27% (n = 15) ward pharmacists. Forty-one participants (74.55 %) indicated there is an AMS programme at their institution of employment while 25.45% (n = 14) indicated they work without an AMS programme.n = 51) agreed and/or strongly agreed on the definition of AMS, median 5 (IQR 5–5). A total of 47 (88.68%) agreed/strongly agreed that an AMS team should consist of a multidisciplinary team including a physician, infectious disease specialist, pharmacist and a microbiologist. Forty-five (84.88%) participants disagreed and/or strongly disagreed that a goal of AMS is to increase the use of antimicrobials while 39 participants (73.58%) agreed and/or strongly agreed that a goal of AMS is to reduce hospital stay. Majority of participants  agreed and/or strongly agreed that optimising antimicrobial use and reducing antimicrobial resistance is a goal of AMS. Participants unanimously agreed and/or strongly agreed that pharmacists play a critical role in AMS to promote optimal use of antimicrobials , educate healthcare professionals , lead and support AMS activities  and to recommend appropriate therapy or de-escalate . p = 0.33; 0.212 and 0.422, respectively). The various current positions of employment showed statistical differences p = 0.015 with ward pharmacists, senior pharmacists, assistant pharmacy managers and clinical pharmacists showing higher levels of participation compared with pharmacists (median 5.4) and pharmacy managers (median 6.2). Participants employed as pharmacists show low levels of participation in monitoring AMS duration (p = 0.015); monitoring antimicrobial consumption (p = 0.13); AMS rounds (p = 0.006); participating in AMS campaigns (p = 0.03) and screening AMS prescriptions in accordance with local guidelines before dispensing, (p = 0.006), medians = 5 compared with clinical pharmacists, ward pharmacists and senior pharmacists. Pharmacists with less than a year of experience showed higher levels of participation  followed by pharmacists with more than 10 years of experience . Pharmacists employed in private sector showed higher participation  compared with those employed in the public sector. Pharmacists working in an institution with an AMS programme demonstrated much higher levels of participation in AMS (median 8) compared with those working without an AMS programme (median 5.2), p = 0.004. Supplementary n = 11; 22%) rated ‘participation won’t make a difference’ as a factor that somewhat limits participation.n = 40; 80%) and least likely to acquire this knowledge in undergraduate programmes . n = 48; 100%).n = 48; 100%). Followed by infectious disease , patient individualisation and surgical prophylaxis . All participants viewed agent of choice as moderately important . Only 28 participants (58.33%) rated standard treatment guideline compliance and team dynamics as a highly important, and 25 participants (52.08%) rated standard treatment guideline compliance as highly important. Supplementary Supplementary Pharmacists have a responsibility to take a prominent role in AMS programmes at institutional level. Previous studies found that a lack of education and training are key barriers to pharmacists’ participation in AMS programmes and activities  compared with only 7.32% in the public sector. Well-established AMS programmes offer a structured and guided approach to AMS activities  were highly rated factor limiting pharmacists’ participation. This could be because of doctors lack of knowledge and understanding about pharmacists’ roles, competencies and activities .Universities worldwide have heeded this call and incorporated AMS training in undergraduate and postgraduate healthcare programmes; however, there is variation in the extent to which AMS training is included, as well as differences in pedagogies (Abubakar & Tangiisuran The study by Burger et al.  further Pharmacy schools should note that pharmacists were all in agreement that a comprehensive course in AMS and infectious disease would be most beneficial if taught in undergraduate programmes. Highly rated topics for inclusion in AMS curricula were infectious disease, dose optimisation, patient individualisation and surgical prophylaxis. The agent of choice was rated as moderately important. In order to be effective members of the AMS team pharmacists require skills and knowledge in antimicrobial use, infectious disease as well as leadership skills to implement AMS programmes and work effectively within a multidisciplinary team (Weier et al. Although it is important to create a solid foundation of AMS knowledge at undergraduate level, AMS education and training should also continue throughout health professional careers to shape and reaffirm their attitudes and behaviours to AMS (Majumder et al. The findings of this study suggest that clinical pharmacists, pharmacists involved in clinical work, and AMS in South Africa have good attitudes, knowledge and perceptions towards AMS and have good levels of participation in AMS activities. Despite the presence of AMS programmes, pharmacist indicated that many factors could limit their level of participation in AMS activities suggesting the need to enhance and develop AMS programmes. Furthermore, the study’s findings confirm that AMS principles are insufficiently incorporated in undergraduate pharmacy programmes. However, informal education and training in the form of short courses, CPD activities, conferences and workshops are available, which pharmacists utilise to improve their knowledge on AMS.There are a few limitations to this study. Firstly, the study was conducted among SASOCP members, clinical pharmacists and pharmacists involved in clinical work and AMS who are not SASOCP members did not get an opportunity to participate in this study. Secondly, there was only a small representation from the public sector. The results from the public sector may differ with greater representation.As this study targeted SASOCP members and had minimum representation from the public sector, further large-scale studies are warranted to validate these results by including a greater number of pharmacists. This study highlighted that pharmacy schools in South Africa incorporate AMS training into their undergraduate curriculum; it is therefore recommended that a study be carried out to determine what pharmacy schools are teaching with regard to AMS. The findings from this study support the recommendation to pharmacy schools to implement systematic education and training for pharmacists, starting from instilling basic principles of AMR, AMS, infectious disease and interpersonal skill development in undergraduate programmes, to continuous education and development in the form of short courses and CPD. The Department of Health should redirect resources to improve AMS programmes in public sectors. Hospital administrators in private and public sectors are encouraged to monitor and support AMS programmes within their institutions. There is strong evidence that doctors are unwilling to work with clinical pharmacists; hence, a study should be conducted to evaluate the factors that influence interprofessional collaboration between doctors and clinical pharmacists.This study found that clinical pharmacists, pharmacists involved in clinical work and AMS in South Africa have good attitudes, knowledge and perception towards AMS. Pharmacists of all positions of employment participate in AMS with pharmacists employed as clinical and ward pharmacist’s showing greater levels of participation. Masters programmes, short courses, CPDs, conferences and workshops are avenues available for pharmacists to acquire AMS knowledge; however, pharmacists are of the view that an undergraduate AMS programme will be most beneficial to bridge the current gap between education and practice.


Recently, astrocytes are fast climbing the ladder of importance in cognitive-related diseases. Perioperative neurocognitive disorder (PND) is a common consequence of anesthesia and surgery, which is widely investigated in elderly and susceptible individuals. There is no doubt that astrocytes also play an irreplaceable role in the pathogenesis of PND. Reactive astrocytes can be found in the PND model, with an altered phenotype and morphology, suggesting a role in the development of the diseases. As a prominent participant cell in the central inflammatory response, the inflammatory response is unavoidably a crucial pathway in the development of the disease. Astrocytes also play a significant role in the homeostasis of the internal environment, neuronal metabolism, and synaptic homeostasis, all of which have an impact on cognitive function. In this article, we discuss the function of astrocytes in PND in order to establish a framework for investigating treatments for PND that target astrocytes. Disordered neurocognitive functions after surgery and anesthesia is a heterogeneous set of situations, which include any form of the acute event (of postoperative delirium) and cognitive decline diagnosed up to 30 days after the procedure (a delayed neurocognitive recovery) and up to 12 months   (p. 11).There are several hypotheses on the pathogenesis of PND. The first, and the most extensively explored, is neuroinflammation  p. 11)..11] .It is of growing interest that astrocytes subject to different injuries develop into distinct subtypes. Astrocytes are classified into A1 and A2 states in the majority of studies; A1 exhibits a detrimental and proinflammatory phenotype, while A2 promotes repair ,23 .oup mice  (p. 13).Presently, many clinical and preclinical studies have confirmed the correlation between neuroinflammation and PND  (p. 13).–C motif ligand 2 (CCL2) plays a vital role in the nervous system and is predominantly derived from astrocytes, while CCR2 (the main receptor for CCL2) is predominantly expressed in microglia. The surgery increases chemokine CCL2 release from the activated astrocytes, which promotes microglia activation and M1 polarization. M1 microglia is one of the phenotypes formed by stimulated microglia, which is considered to be destructive and can produce inflammatory mediators. Cellular interactions promote inflammatory responses and cause hippocampus neuronal injuries, jointly resulting in the development and progression of cognitive impairment [Recent evidence demonstrates that the Cpairment  (p. 13).pairment ,41 .pocampus  (p. 14).pocampus  (p. 14).pocampus  (p. 14).Synapses are specific structures that contact neurons and glial cells in the CNS. Synapses are involved in the transportation of ions and neurotransmitters as well as the exchange of neuronal information  (p. 14).Finally, astrocytes control neurotransmitter transmissions, see 2+-dependent exocytosis, but other mechanisms have also been proposed to mediate the release of glutamate from astrocytes as well [2+ in astrocytes and neuronal excitotoxicity induced by an excessive glutamate release [In contrast, recent studies have revealed that in addition to uptake, astrocytes release trace amounts of glutamate to the adjacent neurons. This release of astrocytic glutamate is plausibly mediated by Ca as well  (p. 14). release  (p. 15).2+ concentrations [2+ influx in hippocampal astrocytes, which leads to cognitive impairment [Acetylcholine is also known to be one of the major neurotransmitters of the central nervous system. Astrocytes in the hippocampus express both nicotinic and muscarinic cholinergic receptors ,63 (p. 1pairment  (p. 15).In addition, D-serine, which interacts with the NMDA receptor’s glycine site and co-activates the NMDA receptor with the agonists  (p. 15),Firstly, astrocytes are a major component of the blood–brain barrier (BBB). Astrocytes can send processes to the vasculature, and these processes terminate with endfeet, which covers up to 98% of the entire vascular surface ,70 . Since chronic isoflurane anesthesia reduces the expression of GJs-Cx43 in the hippocampus, the reduction in GJs-Cx43 levels may further lead to defects in astrocyte networks initiating or exacerbating neuroinflammation and ultimately leading to cognitive impairment in mice  (p. 15).Thirdly, in pathological conditions associated with the BBB breakdown, the endfeet of reactive astrocytes can express tight junction proteins to mount a parenchymal line of defense  (p. 15).Lastly, astrocytes secrete numerous paracrine factors affecting the barrier properties of the BBB, such as morphogens Sonic hedgehog (Shh), vascular endothelial growth factor (VEGF), apolipoprotein E (ApoE) and others ,82 (p. 1Astrocytes represent a major source of apolipoprotein E (ApoE) in the brain. The expression of the human APOE4 isoform is a risk factor for PND  (p. 16).The human brain accounts for only 2% of the overall body weight, but it consumes approximately 20% of energy at rest. Energy expenditure in the brain is boosted even more by various tasks  (p. 16).Firstly, astrocytes regulate cerebral blood flow  (p. 16).In addition to regulating blood flow, astrocytes can transport lactates to neurons, see In addition, astrocytes secrete glial cell line-derived neurotrophic factors (GDNF), which are fundamental in neurogenesis and learning. Amantadine reduces cognitive impairment following anesthesia and surgery by increasing the GDNF level  (p. 17).In summation, astrocytes can influence cognitive performance following anesthesia and surgery via a variety of mechanisms , while s


Precision medicine is a powerful treatment modality for controlling tumor growth. However, given the substantial variability between cancer patients, identifying suitable targets is no easy task. Tumor protein p53 represents one of the most promising anti-cancer drug targets due to its high mutation rate and critical role in tumorigenesis. Despite this, current treatment strategies targeting p53 signaling have only seen modest clinical success presumably due to the complex signaling network surrounding p53 enabling cancer cells to have the ability to adapt to each treatment strategy. This review will focus on the multiple approaches to target p53 and will touch on some of the limitations that may be hindering their clinical success.While chemotherapy is a key treatment strategy for many solid tumors, it is rarely curative, and most tumor cells eventually become resistant. Because of this, there is an unmet need to develop systemic treatments that capitalize on the unique mutational landscape of each patient’s tumor. The most frequently mutated protein in cancer, p53, has a role in nearly all cancer subtypes and tumorigenesis stages and therefore is one of the most promising molecular targets for cancer treatment. Unfortunately, drugs targeting p53 have seen little clinical success despite promising preclinical data. Most of these drug compounds target specific aspects of p53 inactivation, such as through inhibiting negative regulation by the mouse double minute (MDM) family of proteins. These treatment strategies fail to address cancer cells’ adaptation mechanisms and ignore the impact that p53 loss has on the entire p53 network. However, recent gene therapy successes show that targeting the p53 network and cellular dysfunction caused by p53 inactivation is now possible and may soon translate into successful clinical responses. In this review, we discuss p53 signaling complexities in cancer that have hindered the development and use of p53-targeted drugs. We also describe several current therapeutics reporting promising preclinical and clinical results. Despite decades of research and rapidly advancing treatment modalities, cancer remains the second leading cause of premature death worldwide ,2. CellsThe tumor suppressor protein p53 has been called the ‘guardian of the genome’  due to iGenotoxic stress, such as chemotherapy and ionizing radiation, can also activate p53, leading to its accumulation and subsequent transcription of p53-regulated genes due to the convergence of the DNA damage response (DDR) pathway and p53 signaling ,45,46,47ARF in humans and p19ARF in mice) [Cell cycle progression indirectly leads to activation of p53, as transition through the G1 phase to S phase results in the activation of the cyclin-dependent kinases, CDK4 and CDK6, which phosphorylate retinoblastoma protein (pRb) causing dissociation of the pRb–E2F complex. E2F, now free from inhibition, activates the transcription of multiple genes involved in the cell cycle progression from G1 phase to S phase. Interestingly, E2F also facilitates the transcription of ARF (p14in mice) , which iin mice) . Additioin mice) . p53 funin mice) .Activated p53 oligomerizes to form tetramers, which stabilize the binding interaction with p53-response elements in DNA and enable the expression of p53-regulated genes to initiate various cellular programs . p53 regSince its critical role as a tumor suppressor was uncovered ,77,78,79R175H has been found to bind and sequester the tumor suppressors p73 and p63 [+/− and p53−/− mice [p53 is the most mutated protein in cancer, with nearly 50% of all tumor cells displaying some form of p53 mutation , with mu and p63 , both of and p63 . GOF mup−/− mice ,95,96. G−/− mice  and incr−/− mice .p53 is implicated in virtually all the Hanahan and Weinberg “hallmarks of cancer.” Activation of p53 in response to DNA damage, proliferation, and oxidative stress leads to cell cycle arrest and activation of apoptotic pathways . p53 is R172H mutation  did not lead to tumor regression as it does in p53-null mice. Mutant p53R172H was demonstrated to bind to the restored WT p53 and exert a dominant negative effect that prevented the expression of pro-apoptotic proteins but not proteins involved in cell cycle arrest [Tumor suppressors evolved to prevent malignant transformation, with oncogenic stimuli leading to their activation. As such, cancer cells are constantly activating the pathways that evolved to trigger p53 signaling. However, with p53 function lost, they exist in a state of perpetual p53 activation. This led to the hypothesis that if normal p53 function were to be restored, cancer cells would be primed to arrest their growth and undergo apoptosis. Support for this method of treatment was strengthened following reports that replacement of WT p53 into p53-null cell lines resulted in arrested growth, increased apoptosis induction, and caused tumor regression in vivo ,109. p53e arrest . Cancer e arrest ,125. Unfe arrest ,127, ande arrest ,129. A pe arrest ,131. CliDespite the limitations surrounding restorative p53 gene therapy, it does indicate that there is potential that p53 reactivation could result in more meaningful clinical responses if a targeted approach is taken. In the case of cancer cells with WT p53 that present with overexpression of a negative regulator, there are several treatment modalities being explored for each specific indication. Perhaps the most promising and extensively studied are inhibitors of the MDM2-p53 interaction. The discovery that the small molecule nutlin-3a could bind to MDM2 and release p53 from its inhibition resulting in apoptosis and hindered tumor growth in vivo  spurred The variability in therapeutic responses to MDM2 antagonists can be partially explained due to the engagement of multiple p53-dependent pathways apart from apoptosis. Efeyan, A., et al.  demonstrTargeting the MDMX-p53 interaction represents an alternative strategy for restoring p53 activity as it may result in fewer side effects than MDM2 antagonists. Hematopoietic stem cells recover from short-term p53 restoration in mice lacking MDMX, presumably due to functional MDM2 retaining the ability to regulate p53 activity . MDMX tap = 0.13) [Cancer cells possessing mutant forms of p53 can be targeted using compounds that reverse the conformational changes acquired by mutation, thus restoring p53 function. The most clinically advanced compound, PRIMA-1, can preferentially inhibit the growth of cancer cell lines with the R273H DNA contact mutant or the R175H conformation mutant . Upon en = 0.13) . NeitherR175H. Hsp40 can then bind to mup53R175H and stabilize the unfolded protein, restoring its function [Other mup53-selective compounds are in preclinical development. PK7088 targets cancer cells harboring a Y220C p53 mutation, the ninth most common p53 mutation. PK7088 binds mup53 in a pocket created by the Y220C mutation and slows its denaturation . This refunction . The profunction .As if understanding endogenous p53 signaling was not complicated enough, even understanding p53 mutation is not a straightforward topic. Contrary to most tumor suppressor genes that undergo a loss-of-function following mutation, p53 can lose its endogenous function while simultaneously gaining novel tumor-promoting capabilities. DNA contact mutants and conformational mutants alter the DNA binding domain, which can alter the DNA elements recognized by p53 and result in the expression of tumorigenic genes involved in apoptosis resistance, proliferation, angiogenesis, and metastasis . AdditioR248Q slowed tumor growth and prolonged survival. The authors also demonstrated that Hsp90 stabilized mup53, and adding the Hsp90 inhibitor, ganetespib, increased median survival in tumor-bearing mice possessing either the R248Q or the R172H (R175H in humans) p53 mutants by 59% and 48%, respectively, with no effect on p53-null mice. Ganetespib-mediated inhibition of Hsp90 results in selective degradation of mup53 that is accompanied by apoptosis induction, thus providing evidence that elimination of GOF mup53 represents a viable treatment option (In some cases, cancer cells possessing a GOF mutation in p53 are dependent on it for their survival, and knockdown of mup53 abrogates tumorigenic potential in vitro and in vivo ,163. In t option D. Histont option D [164,16t option . PatientPerhaps one of the more obvious strategies to inhibit GOF mup53 would be via knockdown with siRNA. However, there is potential that siRNA may still recognize WT p53 in healthy tissue leading to off-target effects. Martinez et al.  demonstrRather than trying to correct the problems brought on by p53 mutation, an alternative approach is to capitalize on it and utilize the dysfunction for treatment options. The earliest example of this is ONYX-015, a replication-competent adenovirus that selectively infects and lyses cancer cells lacking functional p53. By removing the E1B gene that normally functions to inactivate p53 in infected cells , the adeGOF mutations in p53 can result in unique gene expression signatures relative to WT p53 possessing counterparts . BecauseAn alternative approach for targeting p53 dysfunction that is of particular interest to our group is targeting p53 transcriptional activation ,191. TypVarious oncogenes have been demonstrated to directly bind to the p53 promoter and facilitate its expression, for example, c-Myc  and NF-κRather than targeting specific aspects of p53 inactivation, we propose taking a multifaceted approach to target the p53 network. The Hsp90 inhibitors mentioned above give a great example of this. SAHA and ganetespib facilitate degradation of mup53 protein that abrogates tumorigenic potential in animal models of cancer but failed to perform in the clinic ,166,167.An alternative strategy for targeting the p53 network relies on the use of gene therapy. The WT p53 adenovirus expression vectors have arguably demonstrated the most clinical success of any p53 targeted therapy thus far, but they still face many limitations, primarily low treatment responses and cancer cell adaptation to the WT protein . An addiDespite the complexities of p53 signaling, precision treatment using p53 remains a rapidly growing field in cancer therapy because so many aspects of tumorigenesis are influenced by p53. Unfortunately, despite promising preclinical data, most therapeutic strategies directed toward the modulation of p53 signaling have had low to moderate clinical success. There are a few compounds that show promise though, such as SAHA and ganetespib , that we


We enrolled 133 patients . The median cFGF23 was significantly higher in HD vs. PD patients (p = 0.0017) and not significantly higher in patients without UO (p = 0.12). We found a negative correlation between cFGF23 and the UO volume (p = 0.0250), but the correlation was not significant when considering the type of dialysis treatment. Phosphorus , type of dialysis , and creatinine  were significant and independent predictors of cFGF23 levels. In conclusion, cFGF23 was significantly higher in HD than in PD patients. We found a significant negative correlation between cFGF23 and the residual UO volume, but the correlation was not significant considering the type of dialysis. Our study reveals that dialysis modality is an independent predictor of FGF23 levels. In particular, PD is associated with lower FGF23 levels than HD.Several studies investigated the role of fibroblast growth factor 23 (FGF23) in the regulation of renal phosphate excretion in chronic kidney disease (CKD). However, patients with residual urine output (UO) seem to control their serum phosphorus levels better. Our aim was to determine whether FGF23 levels are influenced by dialysis modality and UO. We performed a cross-sectional study in hemodialysis (HD) and peritoneal dialysis (PD) patients. The C-terminal FGF23 (cFGF23) levels were determined in plasma with a two-site enzyme-linked immunosorbent assay. The UO collection referred to an mL/day measurement. All  Mineral and bone disorders (MBDs) are prevalent in chronic kidney disease (CKD), especially in the advanced stages, representing a severe complication and an important mortality risk factor in both hemodialysis (HD) and peritoneal dialysis (PD) patients .The discovery of the “fibroblast growth factor 23 (FGF23)-bone-kidney axis” has led to several investigations to better understand this complex endocrine network which plays a crucial role in MBD development [FGF23 is a 32 kDa glycoprotein secreted by osteocytes in response to increased levels of 1,25 dihydroxy vitamin D 2D) ,5 and thDespite the fact that no phosphate sensor has yet been identified, most authors agreed that, in CKD, the reduced nephron mass causes a decreased renal phosphate excretion that leads to increased FGF23 secretion from the bone to compensate for hyperphosphatemia . It was The growing attention on FGF23 is partly due to the finding of its correlation with cardiovascular mortality and CKD progression. Concerning cardiovascular risk, a significant correlation was found between the FGF23 level and arterial calcification and stiffness , valve cSome studies have shown that both HD and PD with residual urine output (UO) have better maintenance of phosphate balance than those without UO ,18,19. MOn the other hand, there is no consensus about the phosphate balance in PD and HD patients, as some studies have shown better phosphate control in PD patients ,21, whilThe purpose of the present study was to examine the influence of dialysis type and the presence of residual UO on FGF23 levels.We performed a cross sectional-study in two groups of patients: HD and PD patients. We enrolled patients that underwent HD or PD treatment for at least 3 months with age ≥ 18 years and who agreed to participate in the study between October 2014 and October 2015.Inclusion criteriaAge ≥ 18 years;Patients in chronic dialysis HD or PD (≥3 months);Informed consent was taken before enrollment.Exclusion criteriaDementia;Patients with missing or lacking anamnesis;Hepatic encephalopathy;Heart failure New York Heart Association (NYHA) functional classification IV;Acute myocardial infarction (IMA);Acute coronary syndrome (ACS);Lower respiratory tract infection (LRTI);Systemic inflammatory response syndrome (SIRS);Received immunosuppressive agents;Pregnant.This study was performed with the ethical principles of the Declaration of Helsinki. The protocol was approved by the Ethics Committee for Clinical Trials (CESC) of the province of Vicenza and the ethics committee of San Bortolo Hospital (N.41/14). All participants were informed of the objectives of the study and signed for their informed consent.The data collected from patients were: age (years), gender, hypertension, diabetes mellitus, and causes of CKD classified by 4 categories .Blood samples in EDTA were collected during the outpatient visits in the PD patients or prior dialysis treatment after a long interdialytic interval in HD patients. The plasma was immediately centrifuged and stored at −80 °C.®, SIEMENS Healthcare Diagnostics Inc., and Newark, Tarrytown, NY, USA). Parathyroid hormone (intact PTH) and 25 hydroxy vitamin D (25-(OH)D3) in serum were measured with the chemiluminescent immunoassay, CLIA , Milano, Italy) as routine laboratory exams. We determined the urea, enzymatic creatinine, calcium, and phosphorus (P)  levels were determined in plasma with a two-site enzyme-linked immunosorbent assay, ELISA . Two hundred microliters of plasma were used to assay the samples in duplicates. The samples with values greater than the highest standard were diluted by 1:10 or greater with the 0 pg/mL standard or optional sample diluent and reassayed. The result obtained was multiplied by the dilution factor. The residual urine output (UO) presence was collected from patients, and the quantity of UO (UO volume) was referred to as one day (mL/day).The continuous variables were summarized with the median and range (minimum-maximum) or the mean and standard deviation (±SD) as appropriate, and the categorical ones were summarized with the numbers and percentages of patients in each category.t-test or the Mann-Whitney U test was applied to compare the continuous variables according to their normal or non-normal distribution. The categorical variables were compared with the chi-square or Fisher’s exact test. The correlations between the continuous variables were evaluated with Spearman’s rank correlation (R), and the 95% confidence interval (CI) was calculated using Fisher’s z transformation.All continuous variables were evaluated for normality with a Q-Q plot and with the Shapiro-Wilk test. The student’s p < 0.05 and were considered in a multivariable linear regression model with a backward selection method at the 5% level of significance.A univariable linear regression analysis was used to evaluate the relationship between the FGF23 natural log-transformed and potential predictors. All variables formed statistically significant results with a p-values were two-sided, and the statistical significance was set at p < 0.05. The statistical analyses were performed with SAS 9.4  for Windows.All reported We enrolled 133 patients : 58 HD patients  and 75 PD patients . The principal cause of CKD for all patients  was secondary kidney disease . The residual UO was preserved in 93 (69.9%) patients. The baseline characteristics are summarized in p = 0.0017) (as represented in p = 0.0041) and 25-(OH) D3 (p < 0.0001) were significantly different.The cFGF23 levels were significantly higher in HD compared to PD patients (p = 0.0250)   between cFGF23 and the UO volume ( = 0.56) .p = 0.12).The residual urine output (UO) was preserved in 93 (69.9%) patients. The median cFGF23 level was higher in patients without UO  than in those with a preserved UO  without reaching the statistical significance (p = 0.0983) , there was not a significant difference (p = 0.0005), phosphorus (p = 0.0016), and 25-(OH) D3 (p = 0.0096).The other laboratory variables that resulted in significant differences between patients without and with residual UO were creatinine (p < 0.0001), urea , and phosphorus , and were inversely correlated with UO volume . A significant association was also found with the type of dialysis . The multivariable regression analysis showed that phosphorus , type of dialysis , and creatinine  were significant and independent predictors of cFGF23 levels.To identify the factors associated with cFGF23 levels, we performed a linear regression analysis . In the Mineral and bone disorders are common in the CKD population, especially in the advanced stages; they represent a severe complication and an important risk factor, mortality-related, in patients requiring renal replacement therapy  .Hyperphosphatemia is frequently found in patients with CKD, as the progressive reduction in nephrogenic mass leads to a significant marked decrease in the ability of renal phosphorus excretion (which is taken with diet). FGF23 has a phosphaturic action, as it inhibits the reabsorption of phosphorus at the level of the proximal renal tubule. Hyperphosphatemia results in increased secretion of FGF23 from bone . In advaSome studies have shown that dialysis patients with residual UO have a better phosphorus balance than those without UO ,18,19. CIn our study, 133 patients were enrolled , of which 58 patients had HD  and 75 patients had PD . The residual UO was preserved in 93 (69.9%) of 133 patients. Of the 75 patients with PD, 66 had UO (88%), and of the 58 patients with HD, 27 had UO (46.6%). In our cohort, the percentage of HD patients with a residual UO was higher than the percentage shown in other studies . NonetheThe prevention of hyperphosphatemia in CKD patients using the dietary restriction of phosphate is mandatory. However, dialysis patients have a high risk of protein-energy wasting (PEW) due to multiple causes, such as inflammation, metabolic acidosis, endocrine disturbances, protein losses into the dialysate, and catabolism. Since proteins contain 15 mg of phosphate per gram, a compromise may be necessary, and at least 1.0–1.2 g protein/kg body weight/day is recommended for stable HD and PD patients . MoreoveIn a PD patient, despite an adequate intake of protein (>1 g/kg/day), when their residual renal function decreases, the need for protein decreases.How could we explain why we found more controlled phosphorus values in patients without residual diuresis? Below we have formulated some hypotheses:Some unconsidered aspects in the study, such as compliance with a diet low in phosphorus and adherence to chelation therapy, may have altered the obtained results; it could also be due to the fact that patients with preserved diuresis, precisely for this condition, felt freer (making a mistake) not to follow an adequate diet poor in phosphorus and therefore paradoxically this determined that they had higher median phosphorus values than the dialyzed patients without residual diuresisIn regards to FGF23, the analysis of Bi and colleagues [p = 0.0017)    and not  = 0,12) .The serum FGF23 values in HD patients are higher, probably because compared to the PD patient group, they had a lower percentage of residual UO, although they still had good control of phosphorus. This also explains why the FGF23 values did not change significantly despite the improved phosphorus control.p = 0.0007) proved to be an independent predictor of serum FGF23 levels, and this could be explained by what was already published, i.e., that high values of phosphorus directly stimulate the secretion of FGF23 due to its phosphatic action.In our study, phosphorus (p = 0.0250) (as represented in p = 0.0415). However, the correlation was not significant when considering the type of dialysis   between cFGF23 and the residual UO volume ( = 0.56)  and in tAs also described in the previous work on the re-classification of uremic toxins and their role in CKD , althougThe principle behind the removal of urea toxins in PD is the diffusion of these from the blood through the peritoneal membrane into the peritoneal fluid, which is then eliminated after a certain period of time in the peritoneal cavity. The use of a highly permeable membrane together with better preservation of residual renal function determines that urea toxins are eliminated more efficiently through the peritoneal membrane and that the plasma and tissue levels of these molecules are lower in PD patients than in HD patients . Moreove(1)The cohort was obtained from a current study. Therefore, our studied population was not specifically chosen ad hoc; despite this, we defend the quality of the data obtained as they allowed us to confirm the data from the literature and provided us with points of reflection even for new investigations. What could be especially interesting is obtaining more data about the UO collection;(2)The two groups (patients in HD and patients in PD) were different in number and characteristics;(3)We only used single urinary output data and a single biohumoral examination data point;(4)The compliance with the phosphorus binding therapy and a low phosphorus diet was unknown.In conclusion, cFGF23 was significantly higher in HD patients than in PD patients, and it was higher in patients without residual UO. We found a significant negative correlation between cFGF23 and the residual UO volume, but the correlation was not significant when considering the type of dialysis, as to say that in the relationship between the residual UO and FGF23, the type of dialysis represents a confounding factor. Our study found that the dialysis modality is an independent predictor of FGF23 levels. In particular, PD is associated with lower FGF23 levels than HD.