Most read

All |  Published in last 1 year |  In last 2 years |  In last 3 years
Please wait a minute...
For Selected: Toggle Thumbnails
Identification of appropriate housekeeping genes for quantitative RT-PCR analysis in MDA-MB-231 and NCI-H460 human cancer cell lines under hypoxia and serum deprivation
Amanda P. B. Albuquerque, Meritxell Balmaña, Celso A. Reis, Eduardo I.C. Beltrão
Journal of Molecular and Clinical Medicine    2018, 1 (3): 127-134.   DOI: 10.31083/j.jmcm.2018.03.001
Abstract453)   HTML99)    PDF(pc) (561KB)(148)       Save

Gene expression studies aimed at analyzing cancer cells under hypoxia and serum deprivation conditions show major potential for understanding molecular mechanisms associated with tumor progression as well as resistance to antitumor agents. To the best of our knowledge, a study for the identification of appropriate housekeeping genes in breast and lung cancer cells under hypoxia and serum deprivation conditions is currently missing. Given the relevance of a reliable and accurate normalization, we herein aimed to identify the appropriate housekeeping genes for breast and lung cancer cell lines cultured under hypoxia and/or serum deprivation. The stability of five commonly used housekeeping genes (ACTB, β2M, GUSB, 18S rRNA, and PPIA) was assessed after reverse-transcription quantitative real-time PCR in MDA-MB-231 and NCI-H460 cancer cell lines using GeNorm, NormFinder and BestKeeper software. GeNorm and NormFinder ranking revealed ACTB, GUSB and PPIA as the most stable genes for both tumor cell lines. Our results support the use of ACTB/PPIA for MDA-MB-231 and GUSB/PPIA for NCI-H460 cells as the most stable combination for normalization of gene expression under hypoxic and serum deprivation conditions. Our results highlight the importance of the selection of the housekeeping genes in cancer cells subjected to different physiological stresses, such as hypoxia and serum deprivation.

Table and Figures | Reference | Related Articles | Metrics
Facts and myths of selective casein kinase 1 ε inhibition
Adrián Puerta, Alexis R. Galán, Miguel X. Fernandes, José M. Padrón
Journal of Molecular and Clinical Medicine    2018, 1 (4): 195-200.   DOI: 10.31083/j.jmcm.2018.04.401
Abstract223)   HTML40)    PDF(pc) (501KB)(110)       Save

In the scenario of drug discovery, the challenge is to fully understand and elucidate the mechanism of action to identify, with high resolution, the molecular determinant(s) targeted by the drug and responsible for its pharmacological activity. Cancer offers scientists an almost infinite arena of signaling pathways, targets and small molecules for therapeutic intervention. Among the multiple chemotherapeutic strategies to combat cancer, synthetic lethality remains underexplored. Casein kinase 1 ε (CK1ε) is a serine/threonine protein kinase that has been described as a synthetic lethal partner of the Wnt/β-catenin signaling pathway. Despite its potential as a desirable therapeutic target, only two selective inhibitors are available: PF-4800567 and GSD0054. Until the discovery of GSD0054, CK1ε inhibitors have been considered candidate drugs exclusively in psychopharmacology. In this review, we focus on three key points which we consider essential to define the scope of CK1ε as a synthetic lethal partner and its inhibitors as chemotherapeutics: the therapeutic relevance of this kinase, the scarce availability of selective inhibitors (due to the high homology with its sibling isoform CK1δ ), and the constraint of existing computational tools. This paper represents the first review covering the potential of CK1ε as a druggable target for cancer treatment.

Table and Figures | Reference | Related Articles | Metrics
FAK family kinases in brain health and disease
Kolluru D. Srikanth,Tomer Meirson,Dev Sharan Sams,Hava Gil-Henn
Journal of Molecular and Clinical Medicine    2018, 1 (3): 177-190.   DOI: 10.31083/j.jmcm.2018.03.007
Abstract213)   HTML32)    PDF(pc) (25566KB)(94)       Save

Brain disorders are now identified as one of the largest and costliest health risks throughout human life. While most brain disorders are not life threatening per se, their chronic and incurable nature renders the overall burden from these disorders much greater than would be suggested by mortality figures alone. Several neurodevelopmental conditions, including autism and dyslexia, are being diagnosed at increasing rates throughout the last few decades. Adolescence is now well recognized as a vulnerable brain developmental phase, in which mental disorders such as schizophrenia, depression, and bipolar disorder first appear. Additionally, the constant increase in life expectancy has led to a significant rise in the risk of several neurodegenerative disorders such as Parkinson's disease (PD) and Alzheimer's disease (AD). A primary research goal of neuroscience is to decipher the molecular mechanisms that play direct roles in the pathophysiology of brain disorders, including those of the young and old alike. Research into these mechanisms will have the most significant impact on brain diseases and mental health. The focal adhesion kinase (FAK) and its homologous FAK-related proline-rich tyrosine kinase 2 (Pyk2) define a distinct family of non-receptor tyrosine kinases that are predominantly expressed in the developing as well as in the adult brain. Despite their high similarity, they are believed to fulfill distinct roles within the brain, which are partially determined by their different expression patterns, localization, and interacting proteins. Here, we provide a comprehensive and up-to-date overview of all known neuronal interactors and signaling pathways in which Pyk2 and FAK are involved. Using bioinformatics analysis and statistical tools, we validate, for the first time, the long-term hypothesis by which FAK is involved in axonal guidance and neurodevelopmental signaling, while Pyk2 has a more prominent role in functions of the adult brain, such as memory and learning. We also characterize two new and previously unidentified roles of Pyk2 in neuropathic pain signaling and neuroinflammation. Correlation of the most significant pathways for each kinase with human brain disorders revealed the involvement of Pyk2 in neurodegenerative diseases such as PD, AD, Huntington's disease (HD), and schizophrenia, while FAK was found to be mostly related to neurodevelopmental disorders in which axonal guidance plays a major role, and to a lesser extent to amyotrophic lateral sclerosis (ALS), schizophrenia, mood disorders, and AD. The involvement of FAK in these non-developmental pathways may suggest its possible role in compensating for Pyk2 in specific processes and/or brain disorders. Understanding the molecular mechanisms underlying regulation of FAK family proteins in brain and behavior may lead to novel therapeutic approaches for preventing or treating the underlying causes of neurodevelopmental abnormalities, psychiatric disorders, and neurodegenerative diseases.

Table and Figures | Reference | Related Articles | Metrics
Naphthol-derived Betti bases as potential SLC6A14 blockers
Adrián Puerta, Alexis R. Galán, Roderick Abdilla, Kaylie Demanuele, Miguel X. Fernandes, Giovanna Bosica, José M. Padrón
Journal of Molecular and Clinical Medicine    2019, 2 (2): 35-40.   DOI: 10.31083/j.jmcm.2019.02.7181
Abstract204)   HTML10)    PDF(pc) (1186KB)(55)       Save

Betti bases (aminobenzylnaphthols) have not been studied extensively to explore their possible pharmacological applications. Our group prepared a small and focused library of twenty-three Betti bases from the multicomponent reaction of 2-naphthol with primary or secondary cyclic amines and representative aromatic aldehydes. The compounds were prepared in 52-90% yield using environmentally friendly procedures. The E-factor and the atom economy for our process were 3.92 and 94%, respectively. The study of the anti-proliferative activity against human solid tumor cell lines pointed out that these Betti bases represent privileged scaffolds and could be used for the development of pharmacologically-active compounds in cancer therapeutics. The 50% growth inhibitory (GI50) values of the most potent compounds were in the low micromolar range. Fourteen of these Betti bases were less active in HBL-100 breast cancer cells than towards the breast cancer cell line T-47D. A subset of these Betti bases was further tested against the human breast cancer cell lines MCF-7 and MDA-MB-453. The results indicated a correlation in the sensitivity of T-47D cells to Betti bases. We explored computationally the interaction of the Betti bases with SLC6A14, a Na+- and Cl-- dependent influx transporter of both neutral and cationic amino acids that is overexpressed in T-47D cells. SLC6A14 is inhibited by α-methyl-tryptophan, which blocks cell growth via deprivation of amino acid influx. The docking studies indicated that our Betti bases might behave as tryptophan mimetics, blocking this solute carrier transporter and inducing the anti-proliferative effects. Importantly, these Betti bases showed good cytotoxic selectivity towards cancer cells with no activity against normal human fibroblast cells BJ-hTERT.

Table and Figures | Reference | Related Articles | Metrics
Overcoming Palbociclib Resistance by Combined Treatment with PI3K/AKT/mTOR Inhibitors in Mesothelioma Cells
Graziana Digiacomo, Claudia Fumarola, Daniele Cretella, Roberta Alfieri, Silvia La Monica, Pier Giorgio Petronini, Mara Bonelli, Andrea Cavazzoni
Journal of Molecular and Clinical Medicine    2018, 1 (3): 151-156.   DOI: 10.31083/j.jmcm.2018.03.004
Abstract179)   HTML21)    PDF(pc) (6952KB)(108)       Save

Carcinogenesis of malignant pleural mesothelioma (MPM) is strictly associated with chronic exposure of mesothelial cells from the pleura to asbestos fibers and MPM incidence is expected to peak in the next years. Extensive genome analyses of patient-derived MPM tumors revealed that the most frequent mutational event involves the inactivation of the CDKN2A gene that encodes for the cell cycle inhibitors p16INK4a and p14ARF with consequent constitutive activation of CDK4/6 - cyclin D complexes. Therefore, inhibition of the latter complexes may represent a new option for the treatment of MPM patients. However, despite the efficacy of the specific CDK4/6 inhibitor palbociclib in blocking MPM cell proliferation, acquired resistance inevitably occurs. Herein, palbocilib-resistant clones isolated after stepwise exposure of MSTO-211H cells to gradually increasing drug concentrations, showed a reduction in Rb protein levels as well as in the cell cycle inhibitor p21waf1, accompanied by increased phosphorylation of AKT and p70S6K. Simultaneous treatment of resistant clones with both palbociclib and specific PI3K/AKT/mTOR inhibitors produced an additive effect in terms of reduction in cell growth, without any signs of senescence but with increased cell death. In MSTO-211H sensitive cells, this combination drug treatment significantly delayed the adaptation to palbociclib, suggesting that this treatment approach may prevent or at least retard the emergence of palbociclib resistance. Collectively, these results suggest that the combination of palbociclib with PI3K/AKT/mTOR inhibitors may overcome the acquisition of resistance to palbociclib treatment and could represent a potential therapeutic approach to treat cancers with acquired resistance to CDK4/6 inhibitors in the presence of activation of the AKT/mTOR signaling pathway.

Table and Figures | Reference | Related Articles | Metrics
Lateral Guided Bone Regeneration Using a Novel Synthetic Bioresorbable Membrane: A Two Center Prospective Randomized Controlled Trial Running title: A novel membrane for ridge augmentation
Michal Halperin-Sternfeld, Hadar Zigdon-Giladi, Lior Shapira, Asaf Wilensky
Journal of Molecular and Clinical Medicine    2018, 1 (3): 169-176.   DOI: 10.31083/j.jmcm.2018.03.006
Abstract178)   HTML26)    PDF(pc) (2420KB)(124)       Save

The aim of this study was to evaluate the outcomes of lateral guided bone regeneration (GBR) using a novel resorbable synthetic polyethylene-glycol/methacrylate (PEG/MET) membrane compared to a non-cross-linked collagen membrane (CM). Twenty-eight patients with a potential implant site exhibiting insufficient bone width of ≤ 5 mm were included. Ridge width was measured intraoperatively at 1 mm and 4 mm apical to the crest and via cone-beam computed tomography at baseline and 6 months following GBR using either a PEG/MET or a CM in conjunction with an allograft. During implant placement, core biopsies were harvested and analyzed histomorphometrically. Width changes were calculated. Differences between groups were analyzed using two-sided t-test and Mann-Whitney U-test. The PEG/MET membrane was moldable and exhibited higher strength and stability compared to the CM. Nevertheless, it displayed higher exposure rate of 12/15, compared to 2/13 in the CM sites. At the time of implant insertion, 6 months following GBR, significant gain in bone width was observed in both groups. Mean ridge width at 1 mm and 4 mm apical to the crest was increased significantly from 2.06 ± 0.77 mm and 3.84 ± 1.23 mm to 3.84 ± 1.52 mm and 6.06 ± 2.03 mm (p=0.0006 and p= 0.0009, respectively), with no clinical or radiographic differences between groups. Experimental sites contained more residual scaffold material than the controls (17.4 ± 3.3% and 8.6 ± 2.0%, p= 0.0566). However, bone and connective tissue area fraction were not statistically different between the groups. Overall, despite the higher exposure rate, the new PEG/MET membrane was as successful as a standard collagen membrane in lateral GBR and may have potential use in bone augmentation procedures. This study suggests the feasibility of synthetic membranes, which are not associated with disease transmission, as an attractive alternative to the commonly used CM of bovine or porcine origin.

Table and Figures | Reference | Related Articles | Metrics
Preoperative neutrophil to lymphocyte ratio can improve disease progression prediction of non-muscle invasive bladder cancer
Itamar Getzler, Ofer Nativ, Roy Mano, Jack Baniel, Jacob Rubinstein, Sarel Halachmi
Journal of Molecular and Clinical Medicine    2018, 1 (3): 135-142.   DOI: 10.31083/j.jmcm.2018.03.002
Abstract173)   HTML19)    PDF(pc) (1582KB)(101)       Save

The purpose of this study was prospectively evaluate the ability of Neutrophil-to-Lymphocyte ratio (NLR) to predict disease progression in patients with non-muscle invasive bladder cancer (NMIBC). This is a continuation of our previous retrospective study that indicated the significance of NLR > 2.5 criterion as a predictor of progression in patients with NMIBC. Since December 2013, all patients admitted to Bnai-Zion department for TUR-BT and agreed to participate in the study, had blood analyses for cell count and differential 24hr prior to surgery. Patients with pathological NMIBC were followed prospectively for disease progression. The end-point of the follow up was either a disease progression or the termination of the study. Kaplan-Meier curves and Cox regression were performed to assess the predictive ability of NLR > 2.5 for disease progression. Our results demonstrate a significant difference (p = 0.02) in mean progression-free survival - (35.9 months vs 41.1 months) in the whole cohort Kaplan-Meier survival plot factored by NLR > 2.5. Mean progression-free survival of NLR > 2.5 stratified by stage, grade and treatment (sub-group analysis), showed statistical significance (p = 0.035) for those treated with intra-vesical instillation, and demonstrated a persistent trend for the rest of the stratifications - revealing that the NLR > 2.5 groups always fared worse than the NLR < 2.5 groups. In a univariate analysis, whole cohort Cox regression analysis for disease progression, NLR > 2.5 was found significant (p = 0.05; HR 7.8; CI 1-61), indicating that the probability of progression is increased at least 7-fold for a person with a NLR > 2.5 compared with those with NLR < 2.5. In conclusion, NLR > 2.5 was found to be a significant predictor of disease progression and demonstrated high hazard ratio and worse progression-free survival in patients with NMIBC, especially in those treated with intra-vesical instillation. We propose to consider the incorporation of NLR > 2.5 in the next revisions of the European Organization for Research and Treatment of Cancer (EORTC) scores, given more widely available evidence.

Table and Figures | Reference | Related Articles | Metrics
DPYD Gene Activity Score Predicts Dose-Limiting Toxicity in Fluoropyrimidine-Treated Colorectal Cancer Patients
Dalle Fratte Chiara, Polesel Jerry, Roncato Rossana, De Mattia Elena, Ecca Fabrizio, Bignucolo Alessia, Garziera Marica, Dreussi Eva, Palazzari Elisa, Buonadonna Angela, Guardascione Michela, Berretta Massimiliano, Foltran Luisa, Sartor Franca, D'Andrea Mario, Favaretto Adolfo, Mini Enrico, Nobili Stefania, De Paoli Antonino, Toffoli Giuseppe, Cecchin Erika
Journal of Molecular and Clinical Medicine    2018, 1 (3): 143-150.   DOI: 10.31083/j.jmcm.2018.03.003
Abstract171)   HTML30)    PDF(pc) (305KB)(104)       Save

Pre-treatment DPYD genotyping of a panel of 4-single nucleotide polymorphism (SNP) including DPYD*2A, DPYD*13, c.2846A>T and c.1236A>G-HapB3, has been strongly recommended by the current pharmacogenetics guidelines in order to avoid severe fluoropyrimidine (FL)-related toxicity. However, translation to clinical practice is still lagging behind. This requires a better definition of the relationship between genetic variants of DPYD and dose-limiting toxicities (DLTs) and development of new methods to investigate the effect of DPYD variants, such as the DPYD activity score. The aim of the current study was to support the clinical implementation of DPYD genetic testing, by assessing the relationship between DPYD variants in the 4-SNP panel and the risk to develop DLTs and by stratifying patients according to the DPYD gene activity score (GAS) model. (GAS = 1.0 if carriers of one DPYD*2A or DPYD*13 alleles and GAS = 1.5, if carriers of one c.2846A>T or c.1236G>A-HapB3 allele. Non-carriers GAS = 2.0). A retrospective population of 763 colorectal cancer patients treated with FL-based chemotherapy, was selected and genotyped. Patients carrying at least one decreased function DPYD variant in the 4-SNP panel, displayed a significant association with the risk of developing DLT (i.e. grade ≥ 3 non-hematological toxicity or grade ≥ 4 hematological toxicity) either within the first three cycles of chemotherapy (OR= 2.7, 95% CI = 1.33-5.41) or during the entire course of treatment (OR = 2.7, 95% CI = 1.42-5.04). Patients' GAS was found to better define the risk of DLT for both acute (GAS = 1.5, OR = 1.80, 95% CI = 0.78-4.15 and GAS = 1.0, OR = 10.12, 95% CI = 2.55-40.20) and total toxicity (GAS = 1.5, OR = 2.08, 95% CI = 1.02-4.27 and GAS = 1, OR = 7.09, 95% CI = 1.69-29.65). In conclusion, the present study demonstrates that the DPYD 4-SNP panel and the associated GAS can predict the occurrence of DLT related to treatment with FL. These findings further support the implementation of pre-emptive DPYD genotyping in the routine clinical practice.

Table and Figures | Reference | Related Articles | Metrics
SIRPα is transcriptionally downregulated by epigenetic silencing in medulloblastoma
Dannis G. van Vuurden, Esther Hulleman, Mahban Irandoust, Dennis Biesmans, Timo van den Berg, Eleonora Aronica, Volker Hovestadt, Marcel Kool, W. Peter Vandertop, Gertjan J. L. Kaspers, Jacqueline Cloos
Journal of Molecular and Clinical Medicine    2018, 1 (3): 157-168.   DOI: 10.31083/j.jmcm.2018.03.005
Abstract166)   HTML7)    PDF(pc) (4457KB)(100)       Save

Signal regulatory protein α (SIRPα) is a transmembrane protein that is commonly expressed in cells of the hematopoietic system and brain. Its function is not fully understood but it includes tumor suppressor properties and effects on differentiation. SIRPα may play a role in the development of medulloblastoma (MB), a WHO grade IV brain tumor, which is the most common malignant brain tumor in childhood. The aim of the current study was to determine the possible role of SIRPα in MB cells. Interestingly, in contrast to normal cerebellum, SIRPα mRNA was strongly downregulated in MB and its protein was not detectable in MB tissues. This down-regulation in MB cells was associated with transcriptional silencing of SIRPα via CpG island promoter hypermethylation. Furthermore, Oncomir cluster miR17-92 and miR-106a were correlated with SIRPα gene silencing in MB tumor specimens and cell lines. Histone modification and inhibition of DNA methylation using TSA (20 nM) for 24 hrs and 5-AZA (5 μM) and DZnep (2.5 μM) for 72 hrs, respectively, increased SIRPα expression 25-40 fold and resulted in 90% cytotoxicity of MB tumor cell lines D283-med and D458-med. Remarkably, forced upregulation of SIRPα by viral transduction in MB cell lines did not affect cell growth. In conclusion, SIRPα is epigenetically silenced in MB cells and tumor specimens by promoter hypermethylation and possibly by miRNA expression. SIRPα hypermethylation in MB might reflect the precursor cell state of these cells, rather than being a tumor-specific event, since SIRPα overexpression did not influence MB cell viability. The mechanism of the anti-MB action of epigenetic therapy requires further investigation since our findings indicate that this effect is independent of SIRPα upregulation.

Table and Figures | Reference | Related Articles | Metrics
Adrenergic to mesenchymal fate switching of neuroblastoma occurs spontaneously in vivo resulting in differential tumorigenic potential
Maria C. Lecca, Marianne A. Jonker, U. Kulsoom Abdul, Asli Küçükosmanoglu, Wessel van Wieringen, Bart A. Westerman
Journal of Molecular and Clinical Medicine    2018, 1 (4): 219-226.   DOI: 10.31083/j.jmcm.2018.04.4221
Abstract157)   HTML15)    PDF(pc) (5203KB)(112)       Save

Neuroblastoma is a pediatric tumor that originates from cells of the adrenergic lineage. Here we investigated the balance between differentiation and dedifferentiation in relation to tumor-engraftment potential in preclinical mouse models. We analyzed intratumoral heterogeneity through comparison of marker expression of normal adrenergic development versus tumor marker expression, which showed the presence of sympathoadrenal as well as mesenchymal subtypes of neuroblastoma cells. Subsequently, we evaluated long-term outgrowth capacity of these two (FACS-sorted) cell populations, which showed that adrenergic cells have a stronger long-term clonogenic potential. Engraftment of these sorted populations into mice revealed the occurrence of heterogeneous populations. Modelling of the interconversion rate indicated that cell fate transitions from the adrenergic to mesenchymal state were obtained gradually and stochastically as the tumors grew in mice. We found that adrenergic cells have an increased tumorigenic potential in mice without signs of beneficial cross talk between the two lineage populations. These findings indicate that neuroblastoma contains two rivalling differentiation states that exhibit differences in long term clonal/tumorigenic potential. We expect these states to be relevant for therapy resistance as a result of intratumoral heterogeneity.

Table and Figures | Reference | Related Articles | Metrics
Emissions from a modern log wood masonry heater and wood pellet boiler: Composition and biological impact on air-liquid interface exposed human lung cancer cells
Tamara Kanashova, Olli Sippula, Sebastian Oeder, Thorsten Streibel, Johannes Passig, Hendryk Czech, Tony Kaoma, Sean C. Sapcariu, Marco Dilger, Hanns-Rudolf Paur, Christoph Schlager, Sonja Mülhopt, Carsten Weiss, Carsten Schmidt-Weber, Claudia Traidl-Hoffmann, Bernhard Michalke, Tobias Krebs, Erwin Karg, Gert Jakobi, Sorana Scholtes, Jürgen Schnelle-Kreis, Martin Sklorz, Jürgen Orasche, Laarnie Müller, Ahmed Reda, Christopher Rüger, Anika Neumann, Gülcin Abbaszade, Christian Radischat, Karsten Hiller, Julija Grigonyte, Miika Kortelainen, Kari Kuuspalo, Heikki Lamberg, Jani Leskinen, Ilpo Nuutinen, Tiina Torvela, Jarkko Tissari, Pasi Jalava, Stefanie Kasurinen, Oskari Uski, Maija-Riitta Hirvonen, Jeroen Buters, Gunnar Dittmar, Jorma K. Jokiniemi, Ralf Zimmermann
Journal of Molecular and Clinical Medicine    2018, 1 (1): 23-35.   DOI: 10.31083/j.jmcm.2018.01.004
Abstract155)   HTML14)    PDF(pc) (8586KB)(66)       Save

The consumption of wood fuel is markedly increasing in developing and industrialized countries. Known side effects of wood smoke inhalation manifest in proinflammatory signaling, oxidative stress, DNA damage and hence increased cancer risk. In this study, the composition and acute biological impact of emissions of state-of-the-art wood combustion compliances: masonry heater (MH) and pellet boiler (PB) were investigated. Therefore A549 cells were exposed to emission aerosols in an automated air-liquid interface exposure station followed by cytotoxicity, transcriptome and proteome analyses. In parallel, aerosols were subjected to a chemical and physical characterization. Compared to PB, the MH combustion at the same dilution ratio resulted in a 3-fold higher particle mass concentration (PM2.5) and deposited dose (PB: 27 ± 2 ng/cm2, MH; 73 ± 12 ng/cm2. Additionally, the MH aerosol displayed a substantially larger concentration of aldehydes, polycyclic aromatic hydrocarbons (PAH) or oxidized PAH. Gene ontology analysis of transcriptome of A549 cells exposed to MH emissions revealed the activation of pro-inflammatory response and key signaling cascades MAP kinase and JAK-STAT. Furthermore, CYP1A1, an essential enzyme in PAH metabolism, was induced. PB combustion aerosol activated the proinflammatory marker IL6 and different transport processes. The proteomics data uncovered induction of DNA damage-associated proteins in response to PB and DNA double-strand break processing proteins in response to MH emissions. Taking together, the MH produces emissions with a higher particle dose and more toxic compounds while causing only mild biological responses. This finding points to a significant mitigating effect of antioxidative compounds in MH wood smoke.

Table and Figures | Reference | Related Articles | Metrics
Phenolic peptides as antioxidant and anti-proliferative agents
Azucena Marset-Castro, Álvaro López-Gallardo, Hugo López-Muñoz, Luis Sánchez-Sánchez, Inés Maya, Óscar López, José G. Fernández-Bolaños
Journal of Molecular and Clinical Medicine    2018, 1 (4): 237-248.   DOI: 10.31083/j.jmcm.2018.04.502
Abstract154)   HTML30)    PDF(pc) (2237KB)(70)       Save

We report an efficient synthesis of phenolic peptides starting from 3,4-dihydroxyphenylacetic acid (DOPAC) and L-configured amino acid esters (glycine, phenylalanine, valine, serine, tryptophan, and cystine) using different coupling reagents. The combination of a phenolic scaffold with an amino acid residue might modulate the bioavailability and the therapeutic properties of title derivatives. Moreover, the incorporation of a catechol group, with inherent redox activity, can contribute to alter the redox status of the cancer cells, and therefore, provide anti-proliferative properties. Their activities as antioxidants (i.e. scavenging free radicals and H2O2 as well inhibition of lipid peroxidation) and as anti-proliferative agents against three human cervical carcinoma cell lines (HeLa, ViBo, and CaSki) and normal lymphocytes were evaluated. All compounds exhibited an excellent antioxidant activity; remarkably, the peptide derived from L-cystine exhibited the best antioxidant activity, displaying a 2.5-fold increase in radical-scavenging activity when compared with the natural 2-(3’,4’-dihydroxyphenyl)ethanol (hydroxytyrosol, HT). Moreover, this compound was also the most potent antitumor agent against the three human tumor cell lines (IC50 values 108-122 μM), with a 2-7-fold increase in activity when compared with natural DOPAC and HT, used as reference compounds. Importantly, the cytotoxic activity of these phenolic peptidomimetics against normal human lymphocytes was very low, hence confirming their selectivity towards tumor cells. Moreover, a disulfide-containing peptide also exhibited negligible cell necrosis and a high selectivity against tumor cells when compared to normal lymphocytes. Such derivative incorporates two fragments characterized with redox properties, the catechol moiety, and the disulfide linker. Thus, disulfide-containing phenolic peptidomimetics emerge as good lead candidates for the development of a novel family of anti-tumor agents.

Table and Figures | Reference | Supplementary Material | Related Articles | Metrics
A Small Molecule Tubulin Depolymerizing Agent Identified by a Phenotypic Drug Discovery Approach
Carla Ríos-Luci,Elena Díaz-Rodríguez,Rubén M. Buey,Inês J. Sousa,Miguel X. Fernandes,Atanasio Pandiella,José M. Padrón
Journal of Molecular and Clinical Medicine    2018, 1 (2): 67-76.   DOI: 10.31083/j.jmcm.2018.02.003
Abstract153)   HTML22)    PDF(pc) (2850KB)(124)       Save

In the scenario of drug discovery, numerous in vitro testing initiatives had been established. Thus far, no general methodology is reputable and literature on this hot topic is scarce. In this respect, we propose a strategy based on a Phenotypic Drug Discovery approach. Within our program directed at the discovery of new antitumor agents, we have focused our attention on compounds that disturb the cell cycle. Our strategy relies on the use of a set of biological assays organized in a modular fashion. Herein, we exemplified this strategy with a family of propargylic enol ether derivatives. Using different assays in sequential stages and in a stepwise manner, our studies allowed us to understand the bioactivity of this family of compounds and led us to identify tubulin as the main molecular target.

Table and Figures | Reference | Related Articles | Metrics
Differential mechanism of action of the CK1$\varepsilon $ inhibitor GSD0054
Irene Lagunes, Elva Martín-Batista, Gastón Silveira-Dorta, Miguel X. Fernandes, José M. Padrón
Journal of Molecular and Clinical Medicine    2018, 1 (2): 77-84.   DOI: 10.31083/j.jmcm.2018.02.004
Abstract151)   HTML24)    PDF(pc) (965KB)(103)       Save

In the current study, we explored for the first time, the mechanism of action of the new Casein kinase 1ε (CK1ε) selective inhibitor GSD0054. Although GSD0054 behaved as a selective CKε inhibitor in enzymatic assays, we studied whether this inhibitory activity also occurred inside the cells. The effects of GSD0054 on β-catenin expression and disruption of cell cycle progression were studied in the human breast cancer cell lines MDA-MB-453 (β-catenin negative) and T-47D (β-catenin positive). We also performed molecular modeling studies using computational docking against CK1ε  to explain and predict the mechanism of action of this compound. Moreover, the commercially available CK1ε  inhibitor PF-4800567 and the CK1δ/-ε  inhibitors PF-670462 and IC261 were also studied for comparison purposes.GSD0054 showed anti-proliferative activity against MDA-MB-453 and T-47D cells despite the fact that MDA-MB-453 cells do not possess active β-catenin. However, selective cell killing occurred in the more resistant, β-catenin active, T-47D cells. CK1ε was confirmed as a cellular target, although other targets or alternative mechanisms of action could possibly explain the anti-proliferative activity in MDA-MB-453 cells.

Table and Figures | Reference | Related Articles | Metrics
The whiplash injury phenomenon: a review of the literature
Zvi Gutmacher, Israel Blumenfeld, Rina Elimelech
Journal of Molecular and Clinical Medicine    2018, 1 (4): 249-252.   DOI: 10.31083/j.jmcm.2018.04.405
Abstract151)   HTML16)    PDF(pc) (126KB)(116)       Save

A single traumatic injury to the head, neck and temporomandibular joint that occurs during a sudden motor vehicle collision, is known as whiplash. In spite of the fact that whiplash injury is undefined and does not reflect the biomechanical events of motor vehicle accidents, temporomandibular symptoms may be associated with or occur independently of whiplash-associated disorders. The purpose of this review is to clarify whether a true correlation between temporomandibular joint (TMJ) dysfunc-tion and whiplash injury exists. To this aim, a PubMed/Medline search was conducted using the terms “temporomandibular dysfunction”, “jaw pain”, “temporomandibular joint”, “whiplash”, “motor-vehicle accidents” and “motor-vehicle collisions”. Over 200 related articles were reviewed. The incidence of TMJ dysfunction resulting from whiplash varies from low to moderate and the mechanism of injury is poorly understood. Oral health care providers should be aware of the possible influence of litigation following motor vehicle accidents and its association with the so-called late whiplash syndrome. To date, there is no direct correlation between whiplash injury and TMJ dysfunction. The effect of whiplash on TMJ function is limited in terms of duration and often disappears without complications. A systematic and careful approach is needed when treating TMJ symptoms after whiplash and a differential diagnosis should be considered when temporomandibular joint disorder manifestations occur long after the whiplash incidence.

Reference | Related Articles | Metrics
Novel aspects of the preclinical pharmacology of platinum compounds
Cristina Corno,Paola Perego
Journal of Molecular and Clinical Medicine    2018, 1 (4): 205-212.   DOI: 10.31083/j.jmcm.2018.04.403
Abstract151)   HTML18)    PDF(pc) (2748KB)(109)       Save

Platinum compounds are widely used antitumor agents known to interfere with DNA function by forming DNA crosslinks and DNA-protein crosslinks. Because of their electrophilicity, platinum compounds can interact with nucleophilic residues of all macromolecules. Consequently, this cross-linking inhibits DNA replication in cancer cells. Immunogenic and immunomodulating effects have been ascribed to platinum drugs, with differences and similarities among cisplatin, carboplatin and oxaliplatin. On the one hand, cisplatin is generally unable to induce immunogenic cell death; on the other hand, oxaliplatin appears to be a good inducer, thanks to its capability to efficiently trigger calreticulin exposure to the tumor cell plasma membrane. Conversely, cisplatin, carboplatin and oxaliplatin can relieve immunosuppressive networks e.g., by decreasing PDL-1 and PDL-2 in dendritic and tumor cells. Such drugs are also capable of modulating MHC molecules via IFN-β production and T-cell mediated lysis. The concentrations appear to be key in determining the immunomodulatory properties of these cytotoxic agents, with low in vivo doses usually playing stimulatory effects. As predicted from preclinical models, supportive results have emerged from clinical studies, particularly those based on chemotherapeutic regimens of platinum compounds combined with immunotherapeutics. Future therapeutic interventions are expected to benefit from a better definition of the molecular effects of platinum compounds on the immune system.

Table and Figures | Reference | Related Articles | Metrics
Dimensional changes of the alveolar ridge in the posterior maxilla and sinus pneumatization following teeth extraction
Itay Levi, Michal Halperin-Sternfeld, Hadar Zigdon-Giladi, Eli E. Machtei, Jacob Horwitz
Journal of Molecular and Clinical Medicine    2018, 1 (2): 93-98.   DOI: 10.31083/j.jmcm.2018.02.006
Abstract147)   HTML15)    PDF(pc) (590KB)(162)       Save

Sinus pneumatization is a continuous physiological process that occurs naturally and causes an increase in the volume of paranasal sinuses. Pneumatization is also frequently observed following extraction of teeth in the posterior maxilla. This leads to an increase in the sinus volume and height at the expense of the edentulous alveolar ridge. These changes may affect treatment planning if dental implants are indicated to replace extracted teeth. Using a novel method to align and compare two panoramic radiographs taken before and after tooth extraction, we aimed to examine post-extraction dimensional changes in the maxillary sinus and alveolar ridge by superimposition of preand post-treatment panoramic radiographs. Twenty-two pairs of panoramic radiographs were analyzed retrospectively for changes in alveolar ridge and maxillary sinus dimensions following at least 6 months from tooth extraction. Pre- and post-extraction radiographs were matched and then superimposed using a fixed reference unit. Measurements included the distance from bone-crest to sinus-floor and to sinus-roof, as well as distance from sinus-floor to sinus-roof and maxillary-sinus sagittal circumference. The mean difference between pre- and post-extraction bone-crest to the sinus-floor radiographic measurements was statistically significant (P = 0.001) with a mean change of 1.2 mm. The difference between pre- to post-extraction bone-crest to sinus-roof measurements was insignificant (P = 0.094) with a mean change of 0.9 mm. The distance between pre- and post-extration sinus-floor to sinus-roof was significantly increased in an average of 1 mm (P = 0.001) along with an increase in sinus sagittal circumference from 993.9 ± 295.7 mm to 1096.6 ± 312.5 mm (P < 0.0001). In conclusion, a moderate increase in maxillary sinus dimensions concurrent with crestal resorption may be anticipated after extraction of maxillary posterior teeth, leading to an overall decrease in alveolar bone height.

Table and Figures | Reference | Related Articles | Metrics
Phosphatase Sequencing of Pediatric Acute Myeloid Leukemia Reveals a Novel Mutation in the Phosphatase Gene PTPN4
David GJ Cucchi,Mahban Irandoust,Yehuda G. Assaraf,Gertjan JL Kaspers,Ewart de Bruijn,Mercan Akyuz,Richard AJF Broekhuizen,Nika Heijmans,Erik R Abels,Josta Parigger,Zinia Kwidama,Timo K van den Berg,Valérie de Haas,Dirk Reinhardt,Marry M van den Heuvel-Eibrink,Eveline SJM de Bont,C Michel Zwaan,Edwin Cuppen,Jacqueline Cloos
Journal of Molecular and Clinical Medicine    2018, 1 (1): 1-6.   DOI: 10.31083/j.jmcm.2018.01.001
Abstract145)   HTML17)    PDF(pc) (3186KB)(149)       Save

It has been well established that dysregulated activation of kinases is essential for the development of acute myeloid leukemia (AML). In contrast, little is known about the role of their dephosphorylation counterparts, the tyrosine phosphatases. Here we performed whole tyrosine phosphatome sequencing in 15 pediatric AML samples and found a somatic P394L mutation in the FERM-adjacent region of PTPN4. In the absence of a crystal structure of PTPN4, bioinformatics analysis with the software tool PROVEAN (Protein Variation Effect Analyzer) revealed that this P394L mutation is expected to inflict a deleterious effect on the phosphatase activity of PTNP4. Exploring the frequency of this PTPN4 mutation in 227 acute leukemia samples uncovered an additional silent A364A mutation in exon 13 of PTPN4. No additional mutations were found, which further emphasizes the low mutation burden in pediatric AML. Further functional studies are warranted to explore the actual impact of this P394L mutation on the structure and/or function of PTPN4.

Table and Figures | Reference | Related Articles | Metrics
Low Intensity Pulsed Ultrasound for Accelerating Distraction Osteogenesis: A Systematic Review of Experimental Animal Studies
Jiriys George Ginini,Dekel Shilo,Omri Emodi,Adi Rachmiel
Journal of Molecular and Clinical Medicine    2018, 1 (2): 107-114.   DOI: 10.31083/j.jmcm.2018.02.012
Abstract142)   HTML22)    PDF(pc) (1041KB)(127)       Save

The purpose of this report is to review systematically all studies performed regarding the influence of low intensity pulsed ultrasound (LIPUS) on bone regeneration in animals during distraction osteogenesis (DO). Based on the Preferred Reporting Items for Systematic Reviews and Meta- Analyses (PRISMA) checklist structure, a systematic search using PubMed and EMBASE electronic databases was undertaken utilizing the key words "distraction osteogenesis" and "low intensity ultrasound". Human trials, review articles, case reports and non-English language publications were excluded. Data items were extracted from each eligible study, regarding the study design, risk of bias, results and whether or not LIPUS accelerated bone regeneration. The search identified 40 relevant articles, 15 of which were included for full review. Included studies were characterized by a high risk of bias and considerable variations in study design was observed. However, most studies which reported LIPUS in an intensity of 30-40 mW/cm$^2$ accelerated bone formation via endochondral ossification, thus shortening the consolidation period when applied during distraction and early consolidation periods. According to the current review, application of LIPUS during DO shows promise in accelerating bone formation and density during DO, that bears no adverse effects, thus shortening the consolidation period. Optimal timing of LIPUS application is during the distraction and early consolidation phases. The preferred intensity should be between 30-40 mW/cm$^2$. Histological analysis indicates influence via endochondral ossification. Thus, the effect of LIPUS on chondrocytes should be further investigated in order to decipher the exact molecular and cellular influence of LIPUS on enhancement of bone formation. These findings should be used in future clinical protocols and raise potential directions for future research regarding the molecular mechanisms underlying the influence of LIPUS on bone formation.

Table and Figures | Reference | Related Articles | Metrics
Novel anthracyclines with enhanced immunogenic effects against drug resistant osteosarcoma cells
Elena Gazzano, Joanna Kopecka, Barbara Castella, Ilaria Buondonno, Costanzo Costamagna, Chiara Riganti
Journal of Molecular and Clinical Medicine    2018, 1 (4): 227-236.   DOI: 10.31083/j.jmcm.2018.04.325001
Abstract141)   HTML12)    PDF(pc) (2932KB)(74)       Save

Doxorubicin (dox) is one of the first-line drug treatment in osteosarcoma. P-glycoprotein (Pgp) limits dox’s intracellular accumulation and efficacy in osteosarcoma. Part of the cytotoxic effects of dox are mediated by the induction of immunogenic cell death (ICD) that allows a durable eradication of the tumor by the host immune system. Pgp-overexpressing tumors, however, are also ICD-resistant. We recently synthesized two classes of synthetic doxs - nitric oxide (NO)-releasing dox and H2S-releasing doxthat were cytotoxic against different Pgp-expressing tumors. The aim of this work is to investigate if the lead compounds (termed Ndox and Sdox) were able to elicit ICD in Pgp-positive/dox-resistant osteosarcoma cells. Ndox and Sdox induced apoptosis in both sensitive and resistant cells, were localized within the endolasmic reticulum (ER), up-regulated ER stress-dependent cell death genes, promoted the translocation of calreticulin form ER to cell surface, induced the extracellular release of ATP and HMGB1, increased the phagocytosis of tumor cells by dendritic cells and the expansion of anti-tumor CD8+T-lymphocytes, in a NO- and H2S-dependent manner, respectively. Expanded CD8+clones up-regulated immune-activating cytokines and down-regulated immune-suppressive cytokines. Dox induced the same events in sensitive cells, but not in Pgp-expressing/dox-resistant cells. We suggest Ndox and Sdox as new multifunctional anthracyclines able to induce apoptosis of resistant osteosarcoma cells and contemporarily activate an anti-tumor immune response. These pro-drugs may have a future use in osteosarcoma patients with high Pgp expression, characterized by a poor outcome because of the lack of durable tumor eradication and the high frequency of relapse.

Table and Figures | Reference | Related Articles | Metrics
Semaphorin-3A levels in urine demonstrate promising sensitivity for detection of upper-tract urothelial carcinoma - a preliminary case series
Itamar Getzler,Zahava Vadasz,Jacob Rubinstein,Sarel Halachmi MD
Journal of Molecular and Clinical Medicine    2018, 1 (3): 191-194.   DOI: 10.31083/j.jmcm.2018.03.008
Abstract134)   HTML22)    PDF(pc) (142KB)(119)       Save

The purpose of this study was to evaluate the utility of Semaphorin-3A (Sema-3A) as a biomarker for diagnosis and management of upper tract urothelial carcinoma, independently and in conjunction to cytology. Upper tract noninvasive urothelial carcinoma is a lifelong chronic non-curable disease. It is infamously difficult both to diagnose and to follow-up, and current non-invasive methods are commonly inadequate. Sema-3A protein levels can be measured from a simple urine test and can aid in the diagnosis and follow-up and early non-invasive tumor recurrence detection. Assigned cases for this series were those with pathologically verified upper tract neoplastic lesions. Urine samples for cytology and Sema-3A were taken on admission from all recruited patients. Sema-3A protein levels were determined using ELISA in every sample, and the tumor was graded and staged according to the 2004 WHO grading system. Descriptive and statistical analysis was performed to evaluate the performance of Sema-3A and the cytology exam. This case series included 10 patients with pathologically proven upper tract urothelial carcinoma. Sensitivity for recognizing disease was calculated as 80% for cytology, and 89% for a predetermined Sema-3A cutoff. Combining the strengths of both urine tests to a single criterion (Sema-3A > 5 or positive cytology) resulted in 100% sensitivity. In conclusion, in the current preliminary study, high levels of Sema-3A correlated with upper-tract urothelial cancer stage and displayed higher sensitivity than cytology. Combined analysis of both positive Sema-3A and cytology demonstrated 100% sensitivity. Thus, Sema-3A levels can potentially serve as a reliable biomarker for diagnosis and management for the disease, given that further dedicated studies with larger patient cohorts are undertaken.

Table and Figures | Reference | Related Articles | Metrics
Elucidation of Folate-Mediated Cascades in the Developing Neural Tube: Congenital Malformations Induced by Methyltransferase Inhibition
Robert M, Cabrera, Bogdan J, Wlodarczy k, Richard H. Finnell
Journal of Molecular and Clinical Medicine    2018, 1 (2): 119-126.   DOI: 10.31083/j.jmcm.2018.02.010
Abstract126)   HTML22)    PDF(pc) (500KB)(77)       Save

Folic acid supplementation has been shown to significantly reduce both the occurrence and the recurrence of neural tube defects (NTDs) in human populations, yet the underlying mechanisms for reducing the risk of NTDs continue to be debated. This study examined genetic background and select folate metabolites as possible modifiers that may influence NTD risk. Specifically, several folate cycle and methylation metabolites were examined for their ability to reduce the occurrence of NTDs in two congenic mouse strains carrying targeted disruption of the folate receptor 1 (Folr1) gene. SWV-Folr1tm1Fnn and LM/Bc-Folr1tm1Fnn mice were provided with folate or several folate pathway metabolites, or combinations thereof, to determine the ability of these compounds to rescue nullizygous embryos from lethality and NTDs. Results demonstrated that SWV-Folr1tm1Fnn and LM/Bc-Folr1tm1Fnn mice exhibit different dose responses to folinic acid (5-formyl-tetrahydrofolate) supplementation; however, treating dams throughout gestation with downstream metabolites indicated that only folates rescued Folr1 nullizygous embryos from lethality and NTDs. Chemical inhibitors of folate metabolism were used to further elucidate essential enzymatic and biochemical metabolites. These data demonstrate that the inhibition of S-adenosyl-L-homocysteine hydrolase (AHCY) or selective inhibition of folate responsive isoprenylcysteine carboxylmethyltransferase (ICMT) results in embryo toxicity and fetuses with anterior NTDs. These data indicate that genetic background modifies NTD penetrance in folate-supplemented Folr1tm1Fnn mutants, while downstream metabolites of folate metabolism are not capable of rescuing Folr1tm1Fnn mutants. Moreover, these findings support the hypothesis that the methylation cycle and post-translational methylation of key signaling proteins such as Ras, by ICMT are essential to neural tube closure.

Table and Figures | Reference | Related Articles | Metrics
Chemotherapy induces an immunosuppressive gene expression signature in residual BRCA1/p53-deficient mouse mammary tumors
Sohvi Blatter, Nadine Stokar-Regenscheit, Ariena Kersbergen, Charlotte Guyader, Sven Rottenberg
Journal of Molecular and Clinical Medicine    2018, 1 (1): 7-17.   DOI: 10.31083/j.jmcm.2018.01.002
Abstract119)   HTML15)    PDF(pc) (963KB)(85)       Save

Residual disease is a major hurdle in the eradication of human cancer. We have reproduced this by showing that mammary tumors arising in a mouse model for BRCA1-deficient breast cancer are also not easily eradicated by monotherapy with PARP inhibitors or platinum drugs, despite their high sensitivity to these cytotoxic agents. Tumor regrowth appears to originate from slowly cycling cells with a 2n DNA content that did not enter the S/G2/M phases of the cell cycle during treatment. To identify tumor-intrinsic mechanisms of drug resistance we characterized residual 2n tumor cells by RNAseq in our model. For this purpose, GFP-labelled 2n tumor cells were sorted from residual tumors after cisplatin treatment. We found that these cells display an increased expression of genes encoding immunosuppressive factors including IL-10 and TGF-β, as well as the negative costimulatory signals of the PD-1/PD-L1 and the CTLA-4/B7 axis. By blocking inhibitory T-cell signaling using antibodies directed against CTLA-4 and PD-1 in combination with cisplatin or PARP inhibitors, we attempted at overcoming the immunosuppressive microenvironment in BRCA1-deficient tumors and eradicate residual tumor cells. Expectedly, the combination of CTLA-4 and PD-1-targeting antibodies with the PARP-inhibitor olaparib led to an increase in CD8-positive cytotoxic T-cells in tumor remnants. Remarkably, this increase did not result in a therapeutic benefit and we were not able to eradicate the drug-tolerant tumor cells in vivo using this combination immunotherapy approach. This outcome might be due to other redundant immunosuppressive factors expressed by residual tumor cells as indicated by the increased number of tumor-infiltrating FoxP3-positive T-regulatory cells. Such adverse activation of T-regulatory cells upon immunotherapy may be relevant for the clinic and could explain some of the cases in which CTLA-4 and PD-1-blocking immunotherapy failed.

Table and Figures | Reference | Related Articles | Metrics
Extent of surgical trauma may not be a key factor in Medication-related osteonecrosis of the jaw - a pilot study
Lana Eskander-Hashoul, Rina Elimelech, Zvi Gutmacher, Shai Frankenthal, Doron Rozitsky, Tal Tamari, Hadar Zigdon-Giladi
Journal of Molecular and Clinical Medicine    2018, 1 (2): 85-92.   DOI: 10.31083/j.jmcm.2018.02.005
Abstract116)   HTML18)    PDF(pc) (1972KB)(74)       Save

The pathogenesis of Medication-Related Osteonecrosis of the Jaw (MRONJ) is not fully understood, however, surgical trauma is thought to play a role. Therefore, the aim of the current pilot study was to compare the incidence and characteristics of MRONJ following single or multiple molar tooth extractions in a rat model. To this aim, twenty male Lewis rats were treated with subcutaneous injection of zolendronic acid (ZA), an established bone anti-resorption agent, (7.5 μg/kg) and dexamethasone (Dex), (1 mg/kg), or saline, once a week, for 11 weeks. At three weeks, the first or both first and second maxillary molar teeth were extracted. Eight weeks following extraction, rats were sacrificed and extraction sites were evaluated. Clinical macroscopic examination showed MRONJ-like lesions in all single extraction ZA/Dex-treated rats, showing exposed bone. In the control and multiple extraction ZA/Dex-treated groups, none of the rats showed visible signs of MRONJ. Histological characteristics of MRONJ were found in all ZA/Dex-treated rats (both single and multiple extractions), whereas rats treated with saline showed almost no empty lacunae and necrotic bone. In conclusion, the extent of the surgical field may not be the key factor in MRONJ development since only rats with single tooth extraction displayed exposed bone. However, histological characteristics were identified in both models. Therefore, preclinical studies that aim to evaluate histological features of MRONJ may use both models, whereas when a clinically exposed bone is required, the single tooth extraction model appears to be preferable. Further large scale studies are warranted to corroborate the present findings.

Table and Figures | Reference | Related Articles | Metrics
Re-assembled Casein Micelles for Oral Delivery of Chemotherapeutic Combinations to Overcome Multidrug Resistance in Gastric Cancer
Maya Bar-Zeev, Lotem Nativ, Yehuda G. Assaraf, Yoav D. Livney
Journal of Molecular and Clinical Medicine    2018, 1 (1): 55-65.   DOI: 10.31083/j.jmcm.2018.01.008
Abstract114)   HTML13)    PDF(pc) (5700KB)(92)       Save

Multidrug resistance (MDR) mediated by ATP-dependent efflux transporters continues to be a dominant obstacle towards curative cancer therapy. We have previously demonstrated the ability of β-casein micelles (β-CM) to orally deliver a combination of individually encapsulated hydrophobic cargo of drugs and chemosensitizers designed to overcome MDR in gastric cancer. Herein we investigated the potential of re-assembled casein micelles (rCM) to serve as a target-activated delivery platform comprising a synergistic duo of a chemotherapeutic drug (paclitaxel) and a P-glycoprotein-specific transport inhibitor (tariquidar). The high binding affinity of paclitaxel and tariquidar to rCM was demonstrated by spectrofluorometry. Furthermore, solubilization of the drugs and suppression of crystal growth was shown by light microscopy and dynamic light scattering. A remarkable antitumor activity and complete MDR reversal were demonstrated in the in vitro cytotoxicity assay against MDR human gastric carcinoma cells overexpressing P-glycoprotein. The structure and cytotoxic activity of drug-loaded rCM were completely retained after freeze-drying and reconstitution as in β-CM. Hence, our findings highlight the great potential of casein-based nanovehicles as efficient platforms for oral delivery and local target-activated release of synergistic hydrophobic drug combinations to treat gastric cancer, and overcome of cancer chemoresistance, and for the possible treatment of non-malignant gastric disorders.

Table and Figures | Reference | Related Articles | Metrics
Urothelial cancer rate in women with asymptomatic microhematuria. Is there a justification for full hematuria evaluation?
Dana Shaylovsky Ghersin, Itamar Getzler, Boaz Moskowitz, Ofer Nativ, Sarel Halachmi
Journal of Molecular and Clinical Medicine    2018, 1 (4): 201-204.   DOI: 10.31083/j.jmcm.2018.04.402
Abstract114)   HTML28)    PDF(pc) (362KB)(92)       Save

Background and aim: Microscopic hematuria (MH) is a common finding in urinalysis, existing in up to 30% of evaluated patients. Due to the relatively high rate of malignancy reported in patients with MH, a full urologi-cal evaluation to detect urothelial cancer is advocated by the American Urological Association (AUA) and the European Association of Urology (EAU) in woman with asymptomatic microhema-turia, once infection or urolithiasis were rule out. In contrast to the strict guidelines, our personal experience shows a very low rate of malignancies in women with asymptomatic microhematuria. Hence, the aim of our current study was to assess the rate of urothelial malignancies found during evaluation of asymptomatic women with recurrent MH in a pilot study. Methods: In the current retrospective study we retrieved the records of all women with asymptomatic MH who underwent an elective cystoscopy in our outpatient clinic during the years 2010-2015. We ex-amined the impact of their age, smoking status, upper tract imaging (sonography or CT-Urography), urine cytology results on cystoscopy results and pathological outcome. Results: 165 consecutive patients were included in the study: 1 had abnormal imaging, 2 women had abnor-mal urine cytology (atypia), and 2 had abnormal cystoscopy; 5 women were younger than 35, and all had a completely normal workup. None of the patients were diagnosed with urothelial cancer. Conclusions: A full urological investigation had a low yield in our cohort of women with asymptomatic microhe-maturia, and therefore may be unnecessary, especially in younger women.

Table and Figures | Reference | Related Articles | Metrics
Halofuginone: a novel oral and intravesical agent for the treatment of non-muscle invasive bladder cancer
Omri Nativ, Eilata Dalal, Edmond Sabo, Moshe Aronson, Guy Hidas, Ofer Nativ
Journal of Molecular and Clinical Medicine    2018, 1 (4): 213-217.   DOI: 10.31083/j.jmcm.2018.04.407
Abstract113)   HTML14)    PDF(pc) (478KB)(99)       Save

Background: Non-muscle invasive form (NMIBC) is a chronic disease with a high recurrence rate and requires lifelong surveillance. Various intravesical agents were shown to reduce tumor recurrence but unfortunately, none of these agents proved to be of benefit in long-term prevention of local recurrence or disease progression. Aim of Research: Previous studies have shown that Halofuginone (HF), an antiprotozoal agent, exerts anti-neoplastic activity in various cancer models. Our aim was to evaluate the in vivo activity of oral and intravesical HF treatment in an experimental mouse model harboring NMIBC. Methods: Initially, 60 mice were divided into six treatment groups to evaluate the toxicity of this anti-parasitic agent on the bladder mucosa. The second stage included 126 mice which underwent intravesical implantation with Mouse Bladder Tumor cells (MBT-2): Group 1 (n = 30) received no treatment, group 2 (n = 32) received 6 intravesical instillations of PBS, group 3 (n = 32) received 6 doses of 250 μg oral HF, whereas group 4 (n = 32) received 6 intravesical instillations of 250 μg HF. Results: The average weight of bladders, which reflects the anti-neoplastic activity, differed significantly between the control and treated groups: 88.8 mg ± 15.58 SEM and 81.2 mg ± 13.79 SEM for untreated and PBS-treated mice, respectively, versus 38.0 mg ± 4.02 SEM and 39.6 mg ± 5.97 SEM for animals treated with oral and intravesical HF, respectively. Conclusions: HF exerted a significant anti-neoplastic activity in mice bearing NMIBC upon oral as well as intravesical administration. These results may constitute the basis for the maintenance of oral treatment with HF in patients with NMIBC.

Table and Figures | Reference | Related Articles | Metrics
Retinoids offer new and promising cancer therapeutic avenues
Xin Cao, Quanlin An, Yehuda G. Assaraf, Xiangdong Wang
Journal of Molecular and Clinical Medicine    2019, 2 (2): 23-28.   DOI: 10.31083/j.jmcm.2019.02.9161
Abstract109)   HTML20)    PDF(pc) (407KB)(51)       Save

All trans-retinoic acid (ATRA) as well as several key retinoids including tamibarotene, acyclic retinoid (ACR), and WYC-209, have made a major progress in both pre-clinical cancer therapeutics as well as in the clinical setting regarding the treatment of leukemia and solid tumors via their important impacts on cancer stem cell differentiation or apoptosis. ATRA exerts its antitumor activity by binding to retinoic acid receptors, which in turn specifically bind to DNA as a heterodimer with the retinoid X receptors, at promoter regions known as retinoic acid response elements. The impressive new studies and clinical achievements with retinoids as key pre-clinical research tools and antitumor agents, are summarized and discussed in the current paper. The ongoing clinical trial of tamibarotene, which is the first agent targeting super-enhancers-containing cancers, could provide a new treatment modality for acute myeloid leukemia patients. A recent clinical study for evaluation of the preventive effects of ACR on second primary hepatocellular carcinoma (HCC) demonstrated that the oral administration of ACR for 12 months, significantly reduced HCC recurrence. WYC-209 strongly inhibited cell proliferation of different tumor repopulating cells (TRCs), a highly tumorigenic subpopulation of mouse melanoma cells, and also blocked > 80% of B16 TRCs' lung metastases in wild-type C57BL/6 mice, without any apparent toxicity. These remarkable findings reveal that retinoids constitute a promising class of antitumor agents for the treatment of both hematological malignancies and solid tumors.

Table and Figures | Reference | Related Articles | Metrics
Resistance to crizotinib in a cMET gene amplified tumor cell line is associated with impaired sequestration of crizotinib in lysosomes
Nele Van Der Steen,Richard J Honeywell,Henk Dekker,Johan Van Meerloo,Jeroen Kole,René Musters,Rob Ruijtenbeek,Christian Rolfo,Patrick Pauwels,Godefridus J Peters,Elisa Giovannetti
Journal of Molecular and Clinical Medicine    2018, 1 (2): 99-106.   DOI: 10.31083/j.jmcm.2018.02.007
Abstract106)   HTML9)    PDF(pc) (2908KB)(126)       Save

Several cMET inhibitors have been developed as novel therapeutic candidates and are under investigation in clinical trials. New preclinical models to study mechanisms underlying resistance to these targeted agents are essential, as resistance acquired during treatment may lead to relapse. The squamous non-small-cell lung cancer (NSCLC) cell line EBC-1 harbors a cMET gene amplification and is sensitive to the cMET inhibitor crizotinib. Here, through multiple step selection with gradually increasing concentrations of crizotinib we established a resistant clone of these cells, termed EBC-CR. A tyrosine kinase activity assay did not show increased signaling of a bypassing pathway or renewed activity of cMET after crizotinib treatment. However, the pH-sensitive pHRodo Green AM probe showed increased acidification of the cytoplasm and lysosomes of EBC-CR cells. Live cell fluorescence imaging also showed an increase in lysosomal number after crizotinib treatment, and the intracellular concentration of crizotinib was significantly lower in crizotinib-resistant EBC-CR cells as compared to the drug sensitive parental EBC-1 cells. These findings suggest that the impaired accumulation of crizotinib in EBC-CR cells, together with the increased acidification of the lysosomes, contributes to crizotinib resistance in cMET-amplified NSCLC cells. In conclusion, the present research identified a novel mechanism used by cancer cells to confer resistance to cMET inhibition. These results prompt future studies for the establishment of innovative therapeutic strategies to overcome resistance to cMET kinase inhibitors by modulation of lysosomal acidification.

Table and Figures | Reference | Related Articles | Metrics
A Common Variant in MTHFD1L is Associated with Increased Risk for Spina Bifida
Paige McKenzie,Yunping Lei,Jessica Momb,Dean Appling,Richard H. Finnell
Journal of Molecular and Clinical Medicine    2018, 1 (1): 19-22.   DOI: 10.31083/j.jmcm.2018.01.003
Abstract103)   HTML9)    PDF(pc) (161KB)(120)       Save

Genetic polymorphisms within folate pathway genes represent potentially meaningful risk factors for neural tube defects (NTDs). Here we investigated the association of three previously identified polymorphisms in the folate pathway gene MTHFD1L with spina bifida in a U.S. population consisting primarily of Hispanic and non-Hispanic Caucasians. MTHFD1L allele 1 was found to significantly reduce maternal risk of having a baby with spina bifida, while allele 3 significantly increased maternal risk to have a spina bifida baby. This study confirms that rs3832406 is associated with an increased risk of spina bifida.

Table and Figures | Reference | Related Articles | Metrics
Enhanced Dendritic Cell Development Through Long-Term Proteasome Inhibition
Rieneke van de Ven, Sue Ellen Verbrugge, Marjon Al, Henk Dekker, Henk M.W. Verheul, Janet L. Anderl, Elena T. Chan, Ben A.C. Dijkmans, Willem F. Lems, Rik J. Scheper, Gerrit Jansen, Tanja D. de Gruijl
Journal of Molecular and Clinical Medicine    2018, 1 (1): 37-46.   DOI: 10.31083/j.jmcm.2018.01.005
Abstract103)   HTML6)    PDF(pc) (1618KB)(73)       Save

Bortezomib (BTZ) is a potent and reversible proteasome inhibitor which has shown efficacy in the treatment of multiple myeloma. BTZ also affects NF-κB activity, however, the same mechanisms through which it curtails malignant cell proliferation may also compromise antitumor immunity. As dendritic cells (DC) play a vital role in the elicitation and maintenance of antitumor immunity, we herein studied the long-term effects of BTZ on human DC development and function. The CD34+ MUTZ-3 cell line, stably transduced with human telomerase reverse transcriptase (hTERT), was employed as a sustainable model to study human steady-state DC development and was grown in the presence of gradually increasing BTZ concentrations over a period of 4 months. The resultant BTZ-adapted cells were prospectively assessed for their ability to develop into mature Langerhans cells (LC). Long-term exposure to BTZ (>10 months) at a clinically relevant and apoptosis-inducing concentration (10 nM) provoked spontaneous differentiation in surviving precursors, evidenced by increased expression levels of CD14, TNF receptors and immunoproteasome subunits. Cytokine-dependent differentiation was also enhanced, resulting in increased numbers of mature DC with higher levels of co-stimulatory molecules and an increased capacity for T cell induction. Assessment of nuclear NF-κB subunit levels provided evidence for a role of canonical RelB/p50 activation in the enhanced DC differentiation and maturation established through prolonged BTZ exposure. We conclude that long-term treatment with low dose BTZ is consistent with DC-dependent induction of antitumor immunity.

Table and Figures | Reference | Related Articles | Metrics
The long-term functional impact of post-operative acute kidney injury in patients undergoing nephron-sparing surgery
Zaher Bahouth, Yasmine Ghantous, Edmond Sabo, Omri Nativ, Sarel Halachmi, Boaz Moskovitz, Zaid Abassi, Ofer Nativ
Journal of Molecular and Clinical Medicine    2018, 1 (2): 115-118.   DOI: 10.31083/j.jmcm.2018.02.009
Abstract95)   HTML13)    PDF(pc) (112KB)(66)       Save

Nephron-sparing surgery (NSS) is the standard of care for the management of localized renal tumors. A significant number of patients develop acute kidney injury (AKI) following NSS with potential long-term effect on renal function, and eventually overall survival. The aim of the current study was to assess the long-term functional impact of AKI in patients undergoing NSS. From our NSS cohort, we analyzed the clinical and surgical data of patients treated with NSS. Renal function was assessed using serum Creatinine (sCr) and estimated glomerular filtration rate (eGFR) was estimated using the MDRD equation. SCr was assessed daily starting one day before surgery until discharge. AKI was defined using the latest definition by KDIGO (Kidney Disease: Improving Global Outcomes). Appropriate statistical tests were used to compare between both groups. Of 236 patients, 86 (36.4 %) developed AKI. The vast majority of patients (n = 79) displayed grade I AKI, six had grade II and only one patient had grade III. Mean baseline sCr of the AKI group was 1.11 ± 0.43 mg/dL (median 1.0, range 0.5-3.0), and their long-term mean sCr was 1.4 ± 0.6 mg/dL (median 1.2, range 0.61-4.5). Median follow-up time was 4 years. Most patients (79 %) of the AKI group showed improvement in renal function compared with the immediate post-operative level. However, eighteen patients (21 %) demonstrated stable or deteriorating renal function. The mean difference between last available sCr and baseline sCr in the improved group was 0.12 mg/dL compared to 0.87 mg/dL in the deteriorating group (p = 0.0001). The only statistically significant difference between patients who improved their sCr and patients who did not, was hypertension at diagnosis (p = 0.02).

Table and Figures | Reference | Related Articles | Metrics
The role of developmental signaling pathways in non-small cell lung carcinoma
Darko Durovski, Ornella Randazzo, Godefridus J. Peters, Elisa Giovannetti
Journal of Molecular and Clinical Medicine    2019, 2 (2): 41-53.   DOI: 10.31083/j.jmcm.2019.02.151
Abstract94)   HTML5)    PDF(pc) (777KB)(51)       Save

On a global scale, lung cancer is the most widespread and deadly type of cancer and the non-small cell lung cancer histological subtype, contributes to a significant proportion of this mortality. It has been recently proposed that the main drivers of cancer progression and chemoresistance are cancer stem cells which can be identified through numerous biomarkers or through the overactivation of developmental signaling pathways which are essential for embryonic development but are normally suppressed in adulthood. The primary aim of this review was to compile experimental findings about mediators of three signaling pathways, namely Sonic Hedgehog, Notch and Wingless Integrated, in the prognosis and targeting of three non-small cell lung carcinoma histological types, namely adenocarcinoma, squamous cell carcinoma and large cell neuroendocrine lung carcinoma. Some mediators of all three signaling pathways can be used as biomarkers and overactivation is associated with shorter overall and disease-free survival of patients accompanied by metastasis, epithelial-to-mesenchymal transition and the acquisition of chemoresistance and radioresistance. Additionally, using antagonists to block overexpressed pathway mediators has yielded promising results in vitro with significant apoptotic and anti-tumor activity. Finally, numerous novel mediators of the three pathways have been identified and their pharmacological targeting has resulted in promising pre-clinical findings. The first in-human clinical trials of several drugs are currently being conducted. The current review supports further exploration of the three developmental signaling pathways in the prognosis and targeted treatment of non-small cell lung carcinoma with the aim of enhancing current treatment guidelines with the implementation of targeted therapies.

Table and Figures | Reference | Related Articles | Metrics
ALIX protein analysis: storage temperature may impair results
Lopes-Rodrigues Vanessa,P. R. Xavier Cristina,Sousa Diana,Osório Hugo,G. Assaraf Yehuda,T. Lima Raquel,Helena Vasconcelos M.
Journal of Molecular and Clinical Medicine    2019, 2 (2): 29-34.   DOI: 10.31083/j.jmcm.2019.02.7161
Abstract79)   HTML13)    PDF(pc) (4560KB)(56)       Save

ALIX [ALG-2 (apoptosis-linked gene 2)-interacting protein X] is one of the most well-known molecular biomarkers of extracellular vesicles. Extracellular vesicles are very small vesicles released by most cells and carry in their cargo components from the donor cells, thus being potent vehicles of intercellular (horizontal) communication, influencing various physiological and pathological functions of both recipient and donor cells. The increasing interest in extracellular vesicles highlights the key importance of a reliable analysis of this protein. However, several recent studies in the extracellular vesicles field have shown discrepancies in terms of the expression pattern of apoptosis-linked gene 2-interacting protein X upon Western blot analysis, differing from the theoretical expression pattern of apoptosis-linked gene 2-interacting protein X and its predicted molecular mass. Therefore, to address and clarify this point, we analyzed total protein cell lysates from a chronic myeloid leukemia cell line (K562) for the expression of apoptosis-linked gene 2-interacting protein X by Western blot and mass spectrometry analyses, using protein samples stored at different conditions regarding freezing temperature and storage time. We found that, when stored at -20 oC, a C-terminal specific proteolytic cleavage of apoptosis-linked gene 2-interacting protein X may occur, which depends on the length of storage time. We conclude that analysis of apoptosis-linked gene 2-interacting protein X protein expression should be only carried out when using a wide range of protease inhibitors during isolation of protein cell extract, while preferentially using fresh protein cell extracts or samples that were snap frozen in liquid nitrogen and stored at -80 oC. The current study highlights the importance of proper handling and storage of protein cell lysates for downstream applications in pre-clinical or clinical studies.

Table and Figures | Reference | Related Articles | Metrics
Does PD1/PDL1 Immunotherapy Require Patient Tumor Subgroup Pre-Analysis in the Era of Precision Medicine?
Lucia Scarabel,Giuseppe Toffoli
Journal of Molecular and Clinical Medicine    2018, 1 (1): 51-54.   DOI: 10.31083/j.jmcm.2018.01.007
Abstract71)   HTML5)    PDF(pc) (295KB)(121)       Save

Inter-individual variability of the therapeutic response of patients with cancer to anti-PD1 immunotherapy is a determinant for precision medicine. Anti-PD1 monoclonal antibodies (mAb) interact with the PD1 receptor expressed on T cells, hence preventing the recognition of PDL1 ligand on tumor cells and enhancing their cytotoxic effect. The Food & Drug Administration (FDA) as well as the European Medicines Agency (EMA) have approved anti-PD1 mAb for several human cancer therapies, including malignant melanoma, non-small cell lung cancer, renal cancer, urothelial cancer, and Hodgkin's lymphoma. Anti-PD1 mAb can increase overall survival or progression free survival, but in some subgroups of patients they have shown lower or no therapeutic effect. In recent years, expression levels of PDL1 in tumor cells have been recognized as a determinant for predicting the responsiveness to anti-PD1 mAb therapy, however other factors such as age, or somatic mutations might also play a role. Here we propose the pre-evaluation of PD1 receptor expression status as a prerequisite for patient selection for treatment with anti-PD1 mAb-based therapy.

Table and Figures | Reference | Related Articles | Metrics
Relationship Between Clinical Factors and the Occurrence of Post-Operative Acute Kidney Injury in Patients Undergoing Nephron-Sparing Surgery
Zaher Bahouth, Edmond Sabo, Omri Nativ, Sarel Halachmi, Boaz Moskovitz, Zaid Abassi, Ofer Nativ
Journal of Molecular and Clinical Medicine    2018, 1 (1): 47-50.   DOI: 10.31083/j.jmcm.2018.01.006
Abstract67)   HTML4)    PDF(pc) (132KB)(119)       Save

Nephron-sparing surgery (NSS) is a very common urological procedure performed for clinically localized renal tumors. Compared to radical nephrectomy, it is associated with better renal function preservation and has similar oncologic safety, but with higher complications rate. One of the most important complications is the occurrence of post-operative acute kidney injury (AKI), which has the potential to affect long-term renal function. To study the clinical factors that affect post-NSS renal function and put the patients at risk for AKI. From our NSS cohort, we analyzed the clinical and surgical data of 464 patients who were divided into AKI and non-AKI group. Patients with solitary kidney were excluded. Renal function was assessed using serum Creatinine (sCr) and estimated glomerular filtration rate (eGFR) that was evaluated using the MDRD equation. SCr was assessed daily starting one day before surgery and until the patient was discharged (usually on post-operative day 3). AKI was defined using AKIN and RIFLE criteria. Appropriate statistical analyses were undertaken to compare between the different groups. Of 464 patients, 183 (39.4%) developed AKI following surgery. The AKI patients were more likely to be of male gender (72% vs 58%, p = 0.003), and suffered more from cardiovascular diseases, including hypertension (66.1% vs 52.6%, p = 0.007), ischemic heart disease (24.6% vs 16.3%, p = 0.04) and diabetes mellitus (20.7% vs 13.8%, p = 0.04). Statistically significant differences between the groups in surgical parameters included longer average ischemia time (26 min vs 23.3 min, p = 0.004), higher mean blood loss (153 mL vs 85 mL, p = 0.01), higher transfusion rate (7.1% vs 2.1%, p = 0.01) and the use of tissue adhesive rather than conventional sutures for tumor bed closure, which had a protective effect from AKI. Multivariate analysis showed longer ischemia time, baseline chronic kidney disease and male gender to be the most important and independent risk factors for developing AKI. The most important predictors of AKI are ischemia time, baseline chronic kidney disease and male gender. Of these factors, ischemia time is the only modifiable factor, and hence should be kept to the shortest time possible.

Table and Figures | Reference | Related Articles | Metrics
Risk factors predicting fever following trans-urethral prostatectomy
Kamil Malshy, Omri Nativ, Omer Sadeh, Tareq Aro, Alexander Kastin, Alexander Kravtsov, Gilad E. Amiel, Michael Mullerad, Azik Hoffman
Journal of Molecular and Clinical Medicine    2019, 2 (1): 15-18.   DOI: 10.31083/j.jmcm.2019.01.9231
Abstract44)   HTML7)    PDF(pc) (297KB)(21)       Save

Purpose: Fever occurring post transurethral resection of prostate (TURP) due to urinary tract infection (UTI) is a well-known complication. A better knowledge of pre-operative risk factors will aid the prevention and treatment of this complication. Herein we aimed to describe the risk factors for postoperative fever in a cohort of patients undergoing TURP in a single institute. Methods: A total of 177 patients underwent transurethral resection of prostate (TURP) between January 2016 and August 2017 in our Institute. Postoperative fever was defined as ≥ 38 ℃ up to a week after surgery. Other reasons for post-operative fever were excluded. We collected data for indwelling catheter, antibiotic prophylactic treatment, positive preoperative urine culture, diabetes mellitus (DM), combined cystolithotripsy and prostate size. Results: All patients received antibiotic prophylaxis prior to the TURP procedure. Patients with negative urine culture were treated with IV Amikacin + Ampicillin (n = 83), whereas patients with positive bacterial cultures were treated as per antibiotic sensitivity profile (n = 93). One patient developed fever in the negative culture group, compared to 8 in the positive urine culture group (P = 0.0375). No patient developed septic shock. Smaller prostate size (RR = 1.06, CI-95%, 1.01-1.12, P = 0.016), positive urine culture (RR = 3.85, CI-95%, 1.33-100, P = 0.033) and older age (RR = 1.1, CI-95%, 1.01-1.21, P = 0.031) were all predictors of postoperative fever. In contrast, TURP combined with cystolithotripsy (P = 0.99), indwelling urethral catheter (P = 0.155), and patients with DM (P = 0.256) were not predictors of postoperative fever. Conclusions: Positive urinary bacterial culture prior to TURP is a frequent event. Although positive urinary culture increased the risk of post TURP fever, none of these events caused significant morbidity. Surprisingly, a smaller prostate size increased the risk of postoperative fever. This fact suggests that the degree of infection is attributed to the level of obstruction and not to the volume of the prostate.

Table and Figures | Reference | Related Articles | Metrics
Cryoablation for recurrent renal tumors after primary nephron-sparing surgery using an innovative liquid nitrogen-based cryogenic device
Sagi Shprits, Robert Sachner, Simona Croitoru, Karina Dorfman, Ofir Avitan, Zaher Bahouth, Amnon Zisman, Ofer Nativ
Journal of Molecular and Clinical Medicine    2019, 2 (1): 11-14.   DOI: 10.31083/j.jmcm.2019.01.7261
Abstract39)   HTML6)    PDF(pc) (474KB)(13)       Save

The growing use and improvement of imaging modalities has resulted in increased detection of renal tumors. The gold standard treatment of small renal masses is nephron-sparing surgery. Such surgical intervention is associated with significant rate of local failure. The currently accepted treatment for local failure after nephron-sparing surgery is radical nephrectomy which is associated with very high complication rate as well as loss of functional renal parenchyma. The aim of the current study was to evaluate the safety, technical feasibility, oncologic success and functional preservation of percutaneous cryoablation using a new liquid nitrogen-based cryogenic device in patients with tumor recurrence after nephron-sparing surgery. We present seven patients with tumor recurrence after nephron-sparing surgery who underwent percutaneous cryoablation using ProSenseTM (IceCure Medical Ltd, Caesarea, Israel) under computerized tomography guidance. None of the treated tumor lesions demonstrated contrast enhancement or growth on follow up imaging indicating a 100% oncologic success. Only three adverse events were recorded, all were classified as low grade and resolved spontaneously. In conclusion, percutaneous cryoablation using the novel ProSenseTM device for recurrent renal tumors following nephron-sparing surgery is feasible and effective, with excellent renal function preservation and without major complications.

Table and Figures | Reference | Related Articles | Metrics
A new LDL-masked liposomal doxorubicin overcomes drug resistance in osteosarcoma
Elena Gazzano, Joanna Kopecka, Carolina Dimas Belisario, Costanzo Costamagna, Chiara Riganti
Journal of Molecular and Clinical Medicine    2019, 2 (1): 1-10.   DOI: 10.31083/j.jmcm.2019.01.6161
Abstract33)   HTML4)    PDF(pc) (10252KB)(12)       Save

One of the chemotherapeutic drugs in the first-line treatment of osteosarcoma is doxorubicin (dox). However, its anticancer efficacy is limited by the overexpression of the multidrug efflux transporter P-glycoprotein (Pgp) which extrudes doxorubicin, disrupts drug accumulation and thus hinders the cytotoxic activity of dox The drug extrusion activity of Pgp is enhanced by the presence of cholesterol within the plasma membrane. Pgp expression is up-regulated by multiple transcription factors including hypoxia inducible factor-1α (HIF-1α). Decreasing Pgp expression and drug efflux activity may enhance the efficacy of doxorubicin against drug resistant osteosarcoma. To achieve this goal, we treated two human dox-sensitive cell lines (U-2OS and Saos-2), and their sublines displaying increasing Pgp levels and dox resistance, with a non-toxic dose of atorvastatin. This statin was able to decrease the biosynthesis of farnesyl pyrophosphate (FPP) and cholesterol, two key metabolites that were markedly increased in drug resistant sublines. By reducing the FPP levels, atorvastatin decreased the Ras/ERK1/2/HIF-1α axis and down-regulated Pgp expression. Moreover, by decreasing cholesterol biosynthesis atorvastatin increased the levels of low density lipoprotein receptor (LDLR), that was also progressively increased in the dox-resistant sublines. Taking advantage of this findings, we treated dox-resistant osteosarcoma with atorvastatin followed by treatment with dox encapsulated in liposomes decorated with an LDLR-binding peptide, hence facilitating binding to, and endocytosis via LDLR. This treatment strategy induced high retention of dox in dox-resistant cells, reduced cell viability in vitro and displayed growth inhibition of dox-resistance in mouse xenograft model. Hence, we propose the combination of statins plus LDL-decorated liposomal dox as a novel treatment strategy for Pgp-expressing multidrug resistant osteosarcomas.

Table and Figures | Reference | Related Articles | Metrics
Poly (ADP-ribose) polymerase (PARP) inhibitors as chemosensitizing compounds for the treatment of drug resistant cancers
Silpa Narayanan, Qiu-Xu Teng, Jagadish Koya, Jingquan Wang, Yehuda G. Assaraf, Charles R. Ashby Jr, Zhe-Sheng Chen
Journal of Molecular and Clinical Medicine    2019, 2 (3): 55-67.   DOI: 10.31083/j.jmcm.2019.03.0303
Abstract26)            Save

Poly (ADP-ribose) polymerase (PARP) proteins mediate various cellular processes such as DNA repair, regulation of transcription, protein-protein interaction, expression of inflammatory genes and programmed cell death. PARP proteins have a key role in DNA repair and recent findings have established the role of PARP inhibitors as potent chemotherapeutic drugs. Among the 18 members, PARP1 and PARP2 have been identified as the main targets for the development of pharmacological inhibitors to enhance the cytotoxic efficacy of established anticancer drugs. Furthermore, certain PARP1 and PARP2 inhibitors are being used in combination with other drugs for the treatment of various types of cancer. In different drug resistant cancer cell types, PARP inhibitors have been identified as compounds that reverse the resistance to topoisomerase inhibitors, DNA alkylating and methylating drugs by enhancing the DNA damage induced by these agents. In BRCA mutant cells, with abnormal homologous recombination (HR) repair mechanism, BER (Base Excision Repair Pathway) is responsible for survival of the cells. PARP enzymes play a major role in BER and PARP inhibitors effectively target BRCA mutant cells sparing normal cells via the concept of synthetic lethality, producing minimal toxicity to PARP inhibitors also have a significant role in treating pancreatic adenocarcinoma and castration-resistant prostate cancer. The aim of the current paper is to provide a review on PARP inhibitors and their application in the treatment of various cancer cells which are resistant to standard chemotherapeutic drugs.

Reference | Related Articles | Metrics

Current Issue

  • Volume 2, Issue 2