KRT17 has been described in multi-layered epithelia as well as in tumors derived from these cells. In cancers arising from KRT17 negative single layered epithelia neo-expression of KRT17 has been associated with tumor progression. To obtain more insight into the biology of kidney cancers we have investigated KRT17 expression by immunohistochemistry in normal kidney, in papillary preneoplastic lesions and in 151 papillary and 692 conventional renal cell carcinomas placed on tissue microarray. We found a positive staining in ureteric bud and collecting duct cells in foetal kidney, in all papillary preneoplastic lesions and also in 77% of the 151 papillary renal cell tumors indicating a continuos KRT17 expression during tumor development. The neo-expression of KRT17 in conventional renal cell carcinomas, which derives from KRT17 negative proximal tubules showed a significant correlation with postoperative tumor relapse . In conclusion, the continuous expression of KRT17 from emerging fetal kidney tubules and microscopic pre-neoplastic lesions towards papillary renal cell tumors and its neo-expression in aggressive growing conventional renal cell carcinomas reflects the multiple function of KRT17 in kidney cancers with distinct natural history. This should be taken into account in clinical managements and therapy.Expression of KRT8 and KRT18 in their normal state, but this may change during inflammation, regeneration and also during tumor development. Kidney tubular cells may switch on KRT7 and KRT19, sometimes also KRT17 in addition to the expression of KRT8 and KRT18KRT7, KRT17 and KRT19 appears parallel to the reduction in degree of differentiation. KRT7 and KRT19 are upregulated during renal epithelial injury and repair leading to more plasticity of cells involved in the regeneration processes KRT8 and KRT18 expression to KRT7 and KRT19 is associated with structural remodelling of end stage kidney and increased frequency of tumors expressing both KRT7 and KRT19KRT17 in the biology of kidney cancer is not yet known. Two earlier studies on the keratin expression in distinct types of renal cell cancer using the E3 clone from DAKO did not find KRT17 expression in conventional, chromophobe and papillary renal cell tumors or renal oncocytoma Keratins are intermediate filaments playing a crucial role in the integrity and mechanical stability of single epithelial cells and epithelial tissues. The expression of keratins is regulated in a tissue-type and differentiation-specific manner KRT17 a member of type I acidic epithelial keratin family was first identified in skin basal cell epithelioma and it is considered to be a basal/myoepithelial cell keratin KRT17 occurs in complex, multilayered epithelia and it is largely retained during neoplastic transformation leading to several types of cancer derived from such epithelial cells KRT17 expression has also been detected in several tumors of non-epithelial type but not in their tissue of origin. These studies demonstrated a correlation between neo-expression of the KRT17 and tumor progression KRT17 in fetal development, regeneration and tumorigenesis and on the other hand in the progression of frankly malignant cancers.KRT17 in the biology of renal tumors we applied immunohistochemistry to a large panel of conventional RCC and papillary RCT as well as to preneoplastic lesions (PNL) associated with papillary RCT. In our study we used an antibody arised against the N-terminal protein fragment of KRT17 instead of the E3 clone from DAKO .In attempt to learn about the involvement of We have enrolled tumor samples from consecutively operated patients without selection, who undervent radical or partial nephrectomy due to kidney cancer between 2000 and 2013 at the Department of Urology, Medical School, University of Pecs, Hungary. The only criteria was the availability of clinical data, follow-up and paraffin embedded tumor material. Data on regular follow-up and tumor-specific death were obtained from the Registry of the Department of Urology. Follow-up was defined as time from operation until the last recorded control in 2018, or cancer-specific death. Patients who died from causes other than RCC were excluded from this analysis. Preoperative clinical staging included abdominal and chest computed tomography (CT) scans. Bone scans and brain CT scans were obtained only when clinically indicated. The presence of nodal metastases was confirmed by histological, whereas that of distant metastases by radiographic examination. During the postoperative period patients were surveyed every 6 months by abdominal ultrasound scans, serum creatinine and eGFR measurements, and every year by CT scans. Histological diagnoses were performed by a genitourinary pathologist (GK) according to the Heidelberg and TNM classification systems Haematoxylin and eosin stained slides were reviewed to select the representative paraffin blocks and tumor areas for TMA construction. From each tumor minimum three core biopsies with a diameter of 0.6 mm were placed in the recipient block using a Manual Tissue Arrayer . For marking the TMAs fetal and adult kidney biopsies were included.0C in a moist chamber with rabbit polyclonal anti-KRT17 antibody  at 1:500 dilution. Horse-radish-peroxydase conjugated anti-rabbit, anti-mouse secondary antibody  was applied for 30 minutes at room temperature and the bound antibody was visualized with AEC (Amino-ethyl-carbazol) . Tissue sections were counterstained with Mayer's haematoxylin. In negative control the primary antibody was omitted. The slides were evaluated twice at different times by two of the authors . The staining intensities were scored as low, medium or high. As we did not find substantial differences between weak or strong KRT17 expression and conventional RCC progression, we have evaluated all staining intensity as positive.Paraffin blocks of fetal and adult kidneys, pre-neoplastic lesions and TMAs were used for immunohistochemistry. After deparaffinisation and rehydration the 4 um thick sections were subjected to heat-induced epitope retrieval in citrate buffer, pH 6.0 in 2100-Retriever . Endogenous peroxidase activity and unspecific binding sites were blocked with 3% hydrogen peroxide containing 1% normal horse serum for 15 minutes at room temperature. Slides were incubated overnight at 4KRT17 expression, clinical and pathological parameters were calculated using the χ2 test. The effects of the different variables  on the survival of the patients were estimated by Kaplan-Meier analyses. Comparisons of survival curves were made using the Log rank test. Univariate and multivariate survival analyses were performed using the Cox regression model. Patients that were alive and disease-free were censored. Differences were considered significant at p < 0.05.Data analyses were performed using a SPSS Statistics software package version 25 . Correlations between KRT17 first appeared in the ureteric bud and collecting duct of fetal kidney , the average follow-up was 60.6±33.2 months. Out of the 692 patients 108 (16%) developed metastasis during the follow-up and died due to metastatic cancer. The clinical parameters and correlation with KRT17 expression is shown in Table KRT17 staining which showed a significant correlation with cancer specific death, tumor size, T-stadium, grade and stage. The Kaplan-Meier analysis indicated that patients with KRT17 positive tumor have a significantly shorter survival than those with negative tumor . The KRT17 expression was an independent negative survival factor indicating a nearly three times higher risk of disease relapse and cancer specific death .To estimate the role of KRT17 has other functions as well. KRT17 plays a role in fetal epidermal development and in cell proliferation during wound healing by regulation of protein synthesis through intracellular signaling pathways KRT17 knockout mouse embryos show a delay in closure of surface ectoderm wounds KRT17 binds and sequester TRADD (TNFRSF1A associated via death domain), a mediator of programmed cell death signaling and protects the cells from apoptosis KRT17 null keratinocytes are sensitive to TNF mediated apoptosis KRT17 expression in embryonic ectoderm reflects its commitment towards distinct epithelial lineages and that the program of gene expression characteristic of wound-activated adult keratinocyte may have a relationship to that executed by epithelial cells at specific stages of embryonic development CD44+/KRT17+ cells have stemlike properties, including the capacity for cell renewal and in vivo tumorigenicity Beyond its role to maintain cellular integrity of multi-layered epithelia KRT17 is expressed in simple (one-layered) epithelial cells of fetal and adult kidney. Each PNL of embryonal origin as well as 77% of papillary RCT showed the KRT17 positivity irrespectively of cell type. Taking into account that only 5% of papillary RCTs developed metastasis during 5-year follow-up (data not shown), the role of KRT17 in the progression of papillary RCT can be excluded. Rather, the expression of KRT17 in fetal kidney suggests that it may play a role in the cell specific differentiation of tubular cells similarly to its role in fetal epidermal development KRT17 is associated with the development of PNL and subsequently of papillary RCT. This finding indicates a molecular relationship between kidney development and papillary RCT tumorigenesis. The developmental sequences outlined by KRT17 expression support our hypothesis on the unique natural history of papillary RCT We showed in this study that KRT17 is a basal/myoepithelial cell keratin the expression of which depends on the position of cells in complex multiplayer human epithelia KRT17 has been described in bladder carcinoma, keratoacanthoma and squamous cell carcinoma, oral squamous cell carcinoma, premalignant and malignant squamous lesions of the cervix, all derived from multilayer epithelial cells KRT17 has also been found in several other types of cancer such as gastric adenocarcinoma, ovarian and breast carcinoma, papillary thyroid carcinoma but not in their corresponding normal tissue KRT17 in these cases showed a significant correlation with tumor progression and metastasis. We have identified KRT17 expression in conventional RCCs as an independent negative survival factor indicating a higher risk for postoperative tumour relapse and cancer specific death of patients. Conventional RCC derives from KRT17 negative proximal tubules of the kidney, which suggests that KRT17 has not been involved in the development of conventional RCC. Rather, the neo-expression of KRT17 in conventional RCC can be used as a predictive biomarker similar to gastric, ovarian or breast carcinoma KRT17 in gastric cancer cells inhibited cell proliferation both in vitro and in vivo conditions, reduced the migration of tumor cells and induced apoptosis by BCL2 expression. They also showed that silencing KRT17 leads to cell-cycle arrest at G1/S phase of tumor cells. The clinical, pathological as well as experimental data strongly suggest the involvement of KRT17 neo-expression in tumor progression.Expression of KRT17 is expressed in a single-layered epithelia, and that papillary RCT and conventional RCC have distinct natural history. The continuous expression of KRT17 from fetal kidney tubules towards papillary RCT in adult confirm the relationship between fetal kidney development and tumorigenesis of papillary RCT. And finally, the neo-expression of KRT17 in aggressive growing conventional RCC is significantly associated with postoperative tumor relapse. The multiple functions of KRT17 in kidney cancers unequivocally delineates papillary RCTs and conventional RCCs with distinct natural history, which should be taken into account in clinical managements and therapy.The key novel findings of this study are, that


Ligand binding to death receptors activates apoptosis in cancer cells. Stimulation of death receptors results in the formation of intracellular multiprotein platforms that either activate the apoptotic initiator Caspase-8 to trigger cell death, or signal through kinases to initiate inflammatory and cell survival signalling. Two of these platforms, the Death-Inducing Signalling Complex (DISC) and the RIPoptosome, also initiate necroptosis by building filamentous scaffolds that lead to the activation of mixed lineage kinase domain-like pseudokinase. To explain cell decision making downstream of death receptor activation, we developed a semi-stochastic model of DISC/RIPoptosome formation. The model is a hybrid of a direct Gillespie stochastic simulation algorithm for slow assembly of the RIPoptosome and a deterministic model of downstream caspase activation. The model explains how alterations in the level of death receptor-ligand complexes, their clustering properties and intrinsic molecular fluctuations in RIPoptosome assembly drive heterogeneous dynamics of Caspase-8 activation. The model highlights how kinetic proofreading leads to heterogeneous cell responses and results in fractional cell killing at low levels of receptor stimulation. It reveals that the noise in Caspase-8 activation—exclusively caused by the stochastic molecular assembly of the DISC/RIPoptosome platform—has a key function in extrinsic apoptotic stimuli recognition. Death receptors are targets of novel cancer therapeutics. Most of them signal through flexible multiprotein platforms to either activate apoptotic or necroptotic cell death, or propagate cell survival and pro-inflammatory signals. We focused our study on the role of dynamic assembly and composition of these platforms in the initiation of cell death at the single cell level. Since the assembly is slow through the competitive nature of protein binding within the platforms core we developed a stochastic mathematical model of the death inducing signalling platform. Our model provided an explanation for delayed cell death and fractional killing upon the death receptor stimulation. Additionally, we found that the variability in the cell death response arises through the random assembly initiates a slow noise-prone ramp activation of initiator Caspase-8 spontaneously triggering the apoptotic cascade. Our computational simulations predicted high variation in the time required for cell death induction at the single cell level and highlighted a significant role of death receptor clustering in effective Caspase-8 activation. Our knowledge and data driven model captures detailed processes governing the early events of cell death initiation and can be used to guide the development of more rational combinational treatments against cancer. Apoptotic signalling cascades are designed to irreversibly lead to cell death once specific death thresholds are overcome ,2. Activin vivo studies have shown that fractional death resistance has no direct association with the amount of DRs expressed on the plasma membrane  is the total number of receptors per cell  and Caspase 6 (Casp6) which is triggered by stochastically activated Casp8. Pro-forms of both caspases form stable dimers at physiological concentrations . By cleaAll protein concentrations and parameters used in the model are provided in Tables D and E of Materials and Methods file .in vitro [The estimated weight of the RIPoptosome after short DR-targeted stimulation may exceed 2MDa ,24,29. Tin vitro , howeverThe composition and the time evolution of individual RIPoptosomes within single cell differed from one to another. Consequently, the size and, therefore, molecular weight of those RIPoptosomes varies as well. As an example, we display the composition change in a few randomly chosen RIPoptosomes over the first 20 min with 1 min step interval , S2 Fig.FADD recruitment to the fraction of the high molecular weight complexes is persistently increasing with post treatment time . Our simOn the contrary, ProCasp8 recruitment in the single cell is most abundant in the RIPoptosome of the lower molecular weight . The binMolecular fluctuations in the RIPoptosome composition within single cells cause the fluctuations in the active Casp8 abundance . StochasAveraged over the population the Casp8 activation time course demonstrated high dependence on the dose of the DL as well as the clustering capacity . Thus, eAs expected, the overall variability in the Casp8 activation is a function of the treatment dose . DespiteNext we studied the downstream caspase cleavage cascade, the second deterministic modelling module , which fkcat). Ubiquitination of active Casp3, which is set by XIAP, will attract proteasomal complex leading to Casp3 degradation. However, application of proteasome inhibitors does not stabilise the pool of active Casp3 and consequently does not result in reduced Casp3 proteasomal degradation. Instead, Casp3 catalytic activity is absolutely required for its own proteasomal degradation [kcat needs to be significantly higher (1.75 min-1) from the general  ubiquitin-dependent degradation rate (0.04 min-1) [kcat value    and ODE integration as well as semi-stochastic hybrid model was implemented in the MATLAB 2017b environment see also .S1 Figet al., 2014). Distribution for ligand free (in blue) and ligand bound receptors (in pink) in stimulated cells are followed separately. Distribution of bound receptors was approximated by splines (in red). Distribution for the trimeric Vesicular Stomatitis Virus G protein (VSVG) (in yellow) was used for the peaks calibration. Numbers above the peaks represent amount of monomeric receptor per corresponding cluster and red dots below represent the trimeric receptor complexes. The table demonstrates the conversion of the frequencies into the percentage of clusters from the total DR pool.Experimental frequency distribution of the TNFR1 cluster size on the cellular membrane of unstimulated (black line) and TNFα stimulated HeLa cells derived by Super-resolution PALM microscopy (Fricke (TIF)Click here for additional data file.S2 FigModel generated individual RIPoptosome growth over first 20 min after DR stimulation with 5ng/mL of DL assuming receptor clustering scenario. Plots show eight individual RIPoptosomes randomly chosen from different randomly selected cells.(TIF)Click here for additional data file.S3 FigPopulation average Casp8 activation per cell simulated with model under receptor clustering and disrupted receptor clustering assumption upon stimulation with 5, 7.5, 50 and 250 ng/mL of the DL. Single cell trajectories have been averaged over 100 cells in each represented condition.(TIF)Click here for additional data file.S4 FigAverage FADD abundance per origin simulated for culture of 600 HeLa cells with clustering scenario (A) and without clustering (B) for low (5 ng/mL) and high (50 ng/mL) concentrations of the DL. (C) and (D) represent corresponding quantities for average abundance of ProCasp8 together with DED1-DED2.s.(TIF)Click here for additional data file.S5 FigResult of semi-stochastic model  and full stochastic model  simulation for 20 HeLa cells with clustering scenario for 50 ng/mL concentration of the DL.(TIF)Click here for additional data file.S1 File(PDF)Click here for additional data file.S1 Appendix(M)Click here for additional data file.


Moringa oleifera,” published in this journal  [I read with interest the article by Omabe et al., entitled “Anion Gap Toxicity in Alloxan Induced Type 2 Diabetic Rats Treated with Antidiabetic Noncytotoxic Bioactive Compounds of Ethanolic Extract of Alloxan is one of the oldest chemicals to induce experimental diabetes. It induces type 1 diabetes through a single dose administration in laboratory animals. Alloxan acts in two ways: it selectively inhibits glucose-induced insulin secretion through specific inhibition of glucokinase, the glucose sensor of the beta cell, and it causes a state of insulin-dependent diabetes through its ability to induce reactive oxygen species (ROS) formation, resulting in the selective necrosis of pancreatic beta cells , 3. TherIn the article by Omabe et al., type 2 diabetes has been induced by a single intraperitoneal injection of alloxan, which is not correct due to its mechanism of action.