ARTICLE Download: 1639| View: 7121| Comments: 3 | doi:10.20944/preprints201803.0062.v1
Subject: Medicine & Pharmacology, Pathology & Pathobiology Keywords: Lyme disease; Borrelia burgdorferi; Tickborne disease; Chronic infection; Spirochete culture
Online: 8 March 2018 (07:08:02 CET)
Introduction: Lyme disease is a tickborne illness that generates controversy among medical providers and researchers. One of the key topics of debate is the existence of persistent infection with the Lyme spirochete, Borrelia burgdorferi, in patients who have been treated with recommended doses of antibiotics yet remain symptomatic. Persistent spirochetal infection despite antibiotic therapy has recently been demonstrated in non-human primates. We present evidence of persistent Borrelia infection despite antibiotic therapy in patients with ongoing Lyme disease symptoms. Materials & Methods: In this pilot study, culture of body fluids and tissues was performed in a randomly selected group of 12 patients with persistent Lyme disease symptoms who had been treated or who were being treated with antibiotics. Cultures were also performed on a group of 10 control subjects without Lyme disease. The cultures were subjected to corroborative microscopic, histopathological and molecular testing for Borrelia organisms in four independent laboratories in a blinded manner. Results: Motile spirochetes identified histopathologically as Borrelia were detected in culture specimens, and these spirochetes were genetically identified as Borrelia burgdorferi by three distinct polymerase chain reaction (PCR) methods. Spirochetes identified as Borrelia burgdorferi were cultured from the blood of seven subjects, from the genital secretions of ten subjects, and from a skin lesion of one subject. Cultures from control subjects without Lyme disease were negative for Borrelia using these methods. Conclusions: Using multiple corroborative detection methods, we showed that patients with persistent Lyme disease symptoms may have ongoing spirochetal infection despite antibiotic treatment, similar to findings in non-human primates. The optimal treatment for persistent Borrelia infection remains to be determined.
Fri, 16 November 2018
ARTICLE Download: 278| View: 914| Comments: 0 | doi:10.20944/preprints201811.0395.v1
Subject: Medicine & Pharmacology, Pathology & Pathobiology Keywords: nanopore sequencing; droplet digital PCR; Capnocytophaga canimorsus; diagnosis; sepsis
Online: 16 November 2018 (09:45:38 CET)
We present a case of Capnocytophaga canimorsus septic shock after a dog bite in an immunocompetent individual, where real-time nanopore metagenomic sequencing characterized the microbial agent within 19 hours, with subsequent confirmation using droplet digital PCR. Oral swabs from the dog demonstrated a nearly-identical C. canimorsus isolate by sequencing.
Tue, 18 April 2017
ARTICLE Download: 820| View: 848| Comments: 0 | doi:10.20944/preprints201704.0102.v1
Subject: Medicine & Pharmacology, Pathology & Pathobiology Keywords: aging; age-related disease; mortality rate; positive feedback loop; vicious cycle
Online: 18 April 2017 (02:45:04 CEST)
Age-related diseases (ARDs) are the leading cause of death worldwide, and contribute to 90% of mortality in developed countries. Interestingly, the mortality rates of individual ARDs increase exponentially with age. Processes described by the exponential growth function typically involve a branching chain reaction or, more generally, a positive feedback loop. Here I propose that each ARD is mediated by one or several positive feedback loops (vicious cycles). I then identify critical vicious cycles in five major ARDs: atherosclerosis, hypertension, diabetes, Alzheimer’s and Parkinson’s. I also propose that the progression of ARDs can be halted by selectively interrupting the vicious cycles and suggest the most promising targets. An evolutionary perspective is also offered.
Thu, 16 March 2017
REVIEW Download: 907| View: 795| Comments: 0 | doi:10.20944/preprints201703.0125.v1
Subject: Medicine & Pharmacology, Pathology & Pathobiology Keywords: Extracellular vesicles (EVs); Peptidylarginine deiminases (PADs); Chlor-amidine (Cl-Am); cancer; neurodegeneration; deimination; cytoskeleton; induced pluripotent stem cells (iPSCs); histone H3; epigenetics
Online: 16 March 2017 (17:54:15 CET)
Extracellular vesicle (EV) release, which occurs in most eukaryotic cells, has recently been associated with peptidylarginine deiminase (PAD)-driven protein deimination. Evidence points to the involvement of deiminated cytoskeletal proteins and changes in histone deimination. Both PADs and EVs are associated with various pathologies including cancers, autoimmune and neurodegenerative diseases. The elevated PAD expression observed in cancers may contribute to increase in EV shedding observed from cancer cells, contributing to cancer progression. Similarly, elevated PAD expression observed in neurodegenerative diseases may cause increased EV shedding and spread of neurodegenerative EV cargo, contributing to disease progression and pathologies. Pharmacological inhibition of PAD-mediated deimination using pan-PAD inhibitor Cl-amidine, reduced cellular EV release in prostate cancer cells, rendering them significantly more susceptible to chemotherapeutic drugs. Studies on models of central nervous system damage have demonstrated critical functional roles for PADs and neuroprotective effects using PAD inhibitors in vivo, while human neurodegenerative iPSC in vitro models showed evidence of increased protein deimination. Besides using refined PAD inhibitors to selectively manipulate EV biogenesis for novel combination therapies in cancer treatment, we also speculate how EV biogenesis could be targeted via the newly identified PAD-pathway to ameliorate neurodegenerative disease progression.
Thu, 17 January 2019
REVIEW Download: 103| View: 718| Comments: 0 | doi:10.20944/preprints201901.0170.v1
Subject: Medicine & Pharmacology, Pathology & Pathobiology Keywords: gut microbiota; macrophages; TLR mimicry; immunoepigenetics; metabolism; sterile inflammation
Online: 17 January 2019 (03:05:26 CET)
Perturbation in the microbial population/colony index has harmful consequences on human health. Both biological and social factors influence the composition of the gut microbiota and promote gastric diseases. Changes in the gut microbiota manifest in disease progression owing to epigenetic modification in host which influences differentiation and function of immune cells adversely. Uncontrolled use of antibiotics; chemotherapeutic drugs and change in the diet pattern usually contribute to the changes in the colony index of sensitive strains known to release microbial content in the tissue micromillieu. Ligands released from dying microbes induce TLR mimicry on interaction with TLR abnormally which skew hypoxia and sterile inflammation contributing to severity of disease like IBD autoimmunity and cancer. Various modalities/interventions practiced across the globe and future strategies for microbiota based therapeutic approaches with special emphasis on tumor and inflammatory diseases are reviewed here. Therefore the major aim and scope of this manuscript is to both discuss various modalities/interventions across the globe and to design future microbiota based therapeutic approaches for mitigating the burden with special emphasis on tumor and Inflammatory diseases.
Thu, 13 April 2017
ARTICLE Download: 482| View: 560| Comments: 0 | doi:10.20944/preprints201704.0078.v1
Subject: Medicine & Pharmacology, Pathology & Pathobiology Keywords: Ochratoxin A, insulin, glucagon, glucose, rat plasma, pathology, immunohistochemistry
Online: 13 April 2017 (11:46:19 CEST)
In this study, diabetogenic effects of long term Ochratoxin A (OTA) administration in rats were investigated and its role in the etiology of diabetes mellitus (DM) was examined utilizing 42 female Wistar rats for these purposes. The rats were divided into 3 different study and control groups according to the duration of the OTA administration. Rats received 45 μg OTA daily in their feed for 6, 9 and 24 weeks study groups. Three control groups without any treatment were also used in the same periods. Blood and pancreatic tissue samples were collected during the necropsy at the end of 6, 9 and 24 weeks. Plasma values of insulin, glucagon and glucose in study and control groups were determined. Pancreatic lesions were evaluated by histopathological examination; then insulin and glucagon expression in these lesions were determined by immunohistochemical methods. Statistically significant decrease in insulin levels in contrast to increases in glucagon and glucose levels in blood were observed. Slight degeneration in Langerhans islet cells were observed at the histopathological examination in all OTA treated groups. Immunohistochemistry of pancreatic tissue revealed decreased insulin and increased glucagon expression. This study demonstrated that OTA may cause pancreatic damage in Langerhans islet and predispose rats to DM.
Thu, 18 July 2019
REVIEW Download: 81| View: 560| Comments: 0 | doi:10.20944/preprints201907.0211.v1
Subject: Medicine & Pharmacology, Pathology & Pathobiology Keywords: HIV-1; CRISPR-Cas9; T-cells; lipid nanoparticles; gut-associated-lymphoid tissue; Co-receptors; Probiotics; GI Tract,; Gene Editing
Online: 18 July 2019 (09:51:34 CEST)
HIV-1 is a complicated and perplexing virus. It infects T cells, reverse transcribes its RNA into DNA, utilizes its host DNA machinery to replicate its HIV-DNA, translates the HIV-DNA into proteins, assembles itself for a budding escape from the T cell, and rapidly mutates its conformation. Partially, due to its complexity, there remains no cure for HIV or AIDs. However, recently with the discovery of TALENs, the use of Zinc fingers, and most of all the application of CRISPR-Cas9 technology, has given researchers a new hope in finding alternative gene therapies and treatments for diseases. With more focus on CRISPR-Cas9, this new and novel technology uses a guiding RNA, sgRNA, to lead a Cas9 nuclease to its target for deletion or to change that DNA site. The CRISPR-Cas9 can delete point mutations and multiple DNA sites. Because CRISPR can alter DNA sequences, several scientists have conducted research into CRISPR possibly curing more diseases as cancer, diabetes, and even HIV. HIV-1 drew the focus of a researcher named Dr. Ebina in 2013 when he was the first to design and apply CRISPR-Cas9 to genes found in the binding sites of the HIV-1, inhibiting HIV-1 gene expression. Since 2013 several other researchers have blocked HIV replication and infection through the CRISPR-Cas9 targeting the receptors of T cells called the CC chemokine receptor 5 or CCR5. HIV-1 binds to the CD4 receptor of T cells that consists of co-receptors CCR5 and CXCR4. If CCR5 expression can be removed, the HIV virus can not bind to T-cells, blocking the initial attachment stage, discontinuing the infection. However, there remain obstacles and issues for the CRISPR deletion of CCR5 for treating HIV-1. The issues include: 1) finding new and safe methods of CRISPR-Cas9 delivery, 2) clearing the latent HIV reservoirs, 3) improve the sgRNA design to avoid off-target mutations or deletions, and 4) effectively analyze the viral escape of HIV from CRISPR-Cas9 modifications. Therefore, the purpose of this review is to discuss possible techniques for removing the obstacles that can lessen the potential of CRISPR to delete CCR5, repressing HIV-1 into long-term remission.
Mon, 8 May 2017
REVIEW Download: 559| View: 550| Comments: 0 | doi:10.20944/preprints201705.0071.v1
Subject: Medicine & Pharmacology, Pathology & Pathobiology Keywords: Toxoplasma gondii; neurophysiology; host-parasite interaction, neuroimmune, testosterone, dopamine, catecholamine, glutamatergic
Online: 8 May 2017 (18:34:39 CEST)
Although the parasite Toxoplasma gondii is one of the most pervasive neurotropic pathogens in the world, the host-parasite interactions during CNS infection and consequences of neurological infection are just beginning to be unraveled. The chronic stages of infection have been considered dormant, although several studies have found correlations of infection with an array of host behavioral changes. These may facilitate parasite transmission and impact neurological diseases. During infection, in addition to the presence of the parasites within neurons, host-mediated neuroimmune and hormonal responses to infection are also present. T. gondii induces numerous changes to host neurons during infection and globally alters host neurological signaling pathways, as discussed in this review. Understanding the neurophysiological changes in the host brain is imperative to understanding the parasitic mechanisms and to delineate the effects of this single-celled parasite on health and its contribution to neurological disease.
Tue, 13 June 2017
REVIEW Download: 520| View: 532| Comments: 0 | doi:10.20944/preprints201706.0059.v1
Subject: Medicine & Pharmacology, Pathology & Pathobiology Keywords: mitochondrial; muscle biopsy; ragged red; COX-negative; subsarcolemmal; immunohistochemistry
Online: 13 June 2017 (06:23:36 CEST)
Mitochondria are dynamic organelles ubiquitously present in nucleated eukaryotic cells, subserving multiple metabolic functions, including cellular ATP generation by oxidative phosphorylation (OXPHOS). The OXPHOS machinery comprises five transmembrane respiratory chain enzyme complexes (RC). Defective OXPHOS gives rise to mitochondrial diseases (mtD). The incredible phenotypic and genetic diversity of mtD can be attributed at least in part to the RC dual genetic control (nuclear DNA [nDNA] and mitochondrial DNA [mtDNA]) and the complex interaction between the two genomes. Despite the increasing use of next-generation-sequencing (NGS) and various -omics platforms in unraveling novel mtD genes and pathomechanisms, current clinical practice for investigating mtD essentially involves a multipronged approach including clinical assessment, metabolic screening, imaging, pathological, biochemical and functional testing to guide molecular genetic analysis. This review addresses the broad muscle pathology landscape including genotype-phenotype correlations in adult and paediatric mtD, the role of immunodiagnostics in understanding some of the pathomechanisms underpinning the canonical features of mtD, and recent diagnostic advances in the field.
Wed, 23 August 2017
ARTICLE Download: 530| View: 483| Comments: 0 | doi:10.20944/preprints201708.0080.v1
Subject: Medicine & Pharmacology, Pathology & Pathobiology Keywords: multiple sclerosis; neuroinflammation; astrocytes; myelin; bioinformatics
Online: 23 August 2017 (17:46:28 CEST)
We previously reported that in multiple sclerosis (MS) patients with a progressive form of the disease, spinal cord periplaques extend distance away from plaque borders and are characterized by the co-occurrence of partial demyelination, astrocytosis and low-grade inflammation. However, transcriptomic analyses comparing periplaques to adjacent normal-appearing white matter (NAWM) areas did not allow providing a comprehensive view of molecular events in astrocytes vs oligodendrocytes. Here, we re-assessed our transcriptomic data with the aim of identifying functionally-relevant co-expression networks that would reflect astrocyte vs oligodendrocyte molecular signatures in periplaques. We identified an astrocytosis-related gene module comprising GFAP, the hub gene CX43/GJA1 and a set of transcripts forming a TGFB/SMAD1/SMAD2 genomic signature. Partial demyelination was characterized by a co-expression network which, besides myelin genes, comprised a highly significant number of translation/elongation-related genes. Interestingly, the main oligodendrocyte-related hub we identified was NDRG1, a gene previously shown to be specifically silenced in the NAWM of MS patients. This result indicated that NDRG1 down-regulation could be an important event in the process of periplaque partial demyelination. To establish a putative link between NDRG1 down-regulation and a cytokine/chemokine signature, we then sought to identify cytokine/chemokine genes whose mRNA levels inversely correlated with those of NDRG1. Following this approach we found 5 candidate immune-related genes whose up-regulation associated with NDRG1 down-regulation: TGFB1, PDGFC, IL-17D, IL-33, and IL-12A. From these results we propose that in the spinal cord of MS patients with progressive forms of the disease, TGFB1 may limit acute inflammation but concurrently induce astrocytosis and an alteration of oligodendrocytes terminal differentiation.
Wed, 3 January 2018
ARTICLE Download: 768| View: 386| Comments: 0 | doi:10.20944/preprints201801.0021.v1
Subject: Medicine & Pharmacology, Pathology & Pathobiology Keywords: oxidative stress; in vitro fertilization (IVF); infertile women; follicular fluid (FF); granulosa cells
Online: 3 January 2018 (06:37:39 CET)
Oxidative stress negatively affects folliculogenesis and embryo development. However, a reliable and biologically accurate indicator of oxidative stress does not yet exist. On these bases, the aim of this study was to assess -and compare- blood and follicular fluid (FF) redox status in 45 infertile subjects (and 45 age-matched controls) undergoing in vitro fertilization (IVF) and to establish its connection with the outcome of IVF. Blood and FF were obtained at the time of egg retrieval and immediately analyzed. Firstly, ROS production in blood leukocytes and in granulosa cells was assessed. Oxidative stress markers in blood and in granulosa cells resulted significantly (p<0.001) increased in infertile patients compared to controls. Then, a redox index was obtained in plasma and in FF of patients and controls. The main findings emerging from our study in infertile women are: i) blood oxidative stress reflects FF oxidative stress as demonstrated by the significant correlation between blood redox markers and FF redox markers; ii) a significant correlation between oxidative stress parameters and the considered IVF outcomes is present. We suggest the strict monitoring of the redox parameters for the improvement of assisted reproductive techniques success rate and infertility management.
Wed, 8 August 2018
REVIEW Download: 363| View: 323| Comments: 0 | doi:10.20944/preprints201808.0155.v1
Subject: Medicine & Pharmacology, Pathology & Pathobiology Keywords: amyloid diseases; biocomputing; drug design; natural antiamyloids
Online: 8 August 2018 (04:27:10 CEST)
Amyloids result from the aggregation of several unrelated proteins, due to either specific mutations or promoting intra- or extra-cellular conditions. Structurally, they are rich in intermolecular β-sheets and are the causative agents of several diseases, both neurodegenerative and systemic. It is believed that the most toxic species are small aggregates, referred to as oligomers, rather than the final fibrillar assemblies. Their mechanisms of toxicity are mostly mediated by aberrant interactions with the cell membranes, with resulting derangement of membrane-related functions. Much effort is being put in the search for natural antiamyloid agents, and/or in the development of synthetic molecules. Actually, it is well documented that the prevention of amyloid aggregation results in several cytoprotective effects. Here, we portray the state of the art in the field. Several natural compounds are effective antiamyloid agents, notably tetracyclines and polyphenols. They are generally non-specific, as documented by their partially overlapping mechanisms, and the capability to interfere with the aggregation of several unrelated proteins. Among rationally designed molecules, we mention the prominent examples of β-breakers peptides, whole antibodies and fragments thereof, and the special case of drugs contrasting transthyretin aggregation. In this framework, we stress the pivotal role of the computational approaches. When combined with biophysical methods, in several cases they have helped clarify in detail the protein/drug modes of interaction, which make it plausible that more effective drugs will be developed in the future.
Fri, 25 May 2018
ARTICLE Download: 311| View: 305| Comments: 0 | doi:10.20944/preprints201805.0359.v1
Subject: Medicine & Pharmacology, Pathology & Pathobiology Keywords: autism spectrum disorder; endocrine; estrogen; immune activation; melanin concentrating hormone
Online: 25 May 2018 (10:37:23 CEST)
The voluminous daily output of autism research has become increasingly disconnected, existing largely within highly specific subspecialty areas, and lacking cross-disciplinary linkages of context, theory, and findings to inform a unified body of knowledge. Robust syntheses of published research across the fields of psychiatry, cellular and molecular biology, neurology, endocrinology, immunology, behavioral and social sciences, and pedagogy may help clarify and extend current knowledge by guiding more efficient future research efforts investigating underlying causes, developmental divergences, novel treatments, and specific, sensitive biological markers in autism. This synthesis of interdisciplinary research indicates the hypothalamic-pituitary-adrenal (HPA) stress axis may be at the center of an interaction among sex steroids, immune function, signaling protein transcriptions, neurogenesis, and dysregulation of brain structures sending or receiving projections from the HPA stress axis. These interaction manifest observably as a range of sexually dimorphic behaviors and functional limitations often falling within the current diagnostic features of Autism Spectrum Disorder (ASD). The pathogenicity of endocrine dysregulation may serve as a valuable model for developing a cohesive theory of ASD by explaining how the HPA and connected brain areas respond to extreme conditions of dysregulated endocrine signaling to cause symptoms associated with autism.
Wed, 11 April 2018
ARTICLE Download: 228| View: 250| Comments: 0 | doi:10.20944/preprints201804.0151.v1
Subject: Medicine & Pharmacology, Pathology & Pathobiology Keywords: Cervical cancer – viruses – Human Papillomaviruses – Epstein-Barr virus – Id-1
Online: 11 April 2018 (13:49:57 CEST)
Epstein–Barr virus (EBV) has been recently shown to be co-present with high-risk human papillomaviruses (HPVs) in human cervical cancer; thus, these oncoviruses play an important role in the initiation and/or progression of this cancer. Accordingly, our group has recently viewed the presence and genotyping distribution of high-risk HPVs in cervical cancer in Syrian women; our data pointed out that HPVs are present in 95.45% of our samples. Herein, we aim to explore the co-prevalence of EBV and high-risk HPVs in 44 cervical cancer tissues from Syrian women using polymerase chain reaction (PCR), immunohistochemistry (IHC) and tissue microarray (TMA) analyses. We found that EBV and high-risk HPVs are co-present in 15/44 (34%) of the samples. Additionally, we report that the co-expression of LMP1 and E6 genes of EBV and high-risk HPVs, respectively, is associated with poorly differentiated squamous cell carcinomas phenotype; this is accompanied by a strong and diffused Id-1 overexpression, which is an important regulator of cell invasion and metastasis. These data imply that EBV and HPVs are co-present in cervical cancer in the Middle East area including Syria and their co-presence is associated with a more aggressive cancer phenotype. Future investigations are needed to elucidate the exact role of EBV and HPVs cooperation in cervical carcinogenesis.
Fri, 28 December 2018
REVIEW Download: 90| View: 218| Comments: 0 | doi:10.20944/preprints201812.0333.v1
Subject: Medicine & Pharmacology, Pathology & Pathobiology Keywords: gap junction; connexin; angiogenesis; cell mechanics; cell migration; cellular stiffness
Online: 28 December 2018 (05:19:52 CET)
Angiogenesis, the sprout and growth of new blood vessels from existing vasculature, is an important process of tumor development for the supply of oxygen and nutrition to cancer cells. Endothelial cell is a critical player in angiogenic process by modulating cell proliferation, cell motility, and cell morphology in the response to pro-angiogenic factors and environments provided by tumor and cancer cells. Recent in vivo and in vitro studies have revealed that gap junction of endothelial cells also participates in the promotion of angiogenesis. Pro-angiogenic factors modulate gap junction function and connexins expression in endothelial cells, whereas endothelial connexins involve in angiogenic tube formation and cell migration of endothelial cells via both gap junction channel function dependent or independent mechanisms. In particular, connexin might have the potential to regulate cell mechanics such as cell morphology, cell migration, and cellular stiffness that are dynamically changed during angiogenic processes. Here, we review the implication for endothelial gap junction and cellular mechanics in vascular angiogenesis.
Fri, 23 August 2019
REVIEW Download: 94| View: 214| Comments: 0 | doi:10.20944/preprints201908.0234.v1
Subject: Medicine & Pharmacology, Pathology & Pathobiology Keywords: osteoarthritis; articular cartilage; degeneration; regeneration; therapeutic protein; growth factor; protein production platform; protein packaging cell line; transforming growth factor β (TGF-β); GP2-293 cells
Online: 23 August 2019 (03:33:49 CEST)
This article focuses on the current state-of-the-art in the area of cellular and molecular biotechnology for over-production of clinically relevant therapeutic growth factors and how the technology can be used for the treatment of osteoarthritis (OA). Transfected and irradiated protein packaging cell lines may be used as “cellular factories” for large-scale production of therapeutic proteins and pro-anabolic growth factors, particularly in the context of cartilage matrix regeneration. We discuss the potential for new innovations in regenerative medicine for degenerative diseases of synovial joints using mammalian protein production platforms, specifically protein packaging cell lines, for over-producing growth factors for cartilage tissue regeneration and give recent examples. Mammalian protein production platforms that incorporate protein packaging cell lines are superior to bacterial expression systems and are likely to have a significant impact on the development of new biological therapies for treating focal cartilage defects and more generally for the treatment of degenerative joint diseases such as OA.
Wed, 10 October 2018
ARTICLE Download: 117| View: 213| Comments: 0 | doi:10.20944/preprints201810.0211.v1
Subject: Medicine & Pharmacology, Pathology & Pathobiology Keywords: lipoprotein; extracellular vesicles; exosome; ectosome; stress response; resistant cancer; metastatic cancer; heat shock stress
Online: 10 October 2018 (09:44:17 CEST)
Resistant cancer often shows a particular secretory trait such as heat shock proteins (HSPs) and extracellular vesicles (EVs), including exosomes and oncosomes surrounded by lipid bilayers. Lipoproteins are biochemical assemblies that transport hydrophobic lipid (a.k.a. fat) molecules in body fluid and are composed of a single-layer phospholipid and cholesterol outer shell, lipids molecules within the particles, and apolipoproteins embedded in the membrane. However, lipoprotein storage and secretion by cancer cells have not well-investigated yet. We found lipoproteins were stored and abundantly secreted by neuroendocrine, castration-resistant prostate cancer (NEPC / CRPC) cells but barely secreted by colon cancer cells and oral squamous cell carcinoma (OSCC) cells. In addition, large EVs (approx. 300 nm diameter) and potential oncosomes were released by CRPC and OSCC cells. Proteomics revealed that CRPC cells secreted EVs enriched with tetraspanins and extracellular matrices which were reduced upon heat shock stress and alternatively lipoproteins and HSPs were secreted upon stress. Heat shock stress triggered secretion of lipoprotein-EV complexes that contained apolipoprotein A, B, C and E. These data suggested that vesicular assembly composed of EVs and lipoproteins enriched with cholesterols and phospholipids may be stored in resistant cancer cells but released upon cell stress that is increased in cancer therapies.
Tue, 13 November 2018
ARTICLE Download: 119| View: 209| Comments: 0 | doi:10.20944/preprints201811.0310.v1
Subject: Medicine & Pharmacology, Pathology & Pathobiology Keywords: tessellation; forensic picture; bullets; homology; proximity
Online: 13 November 2018 (10:41:14 CET)
Here we show how a recently-introduced method from algebraic topology, namely proximal planar vortex 1-cycles, might be helpful in detecting hidden features of the shapes and holes in images, therefore contributing to the solution of both cold and fresh forensic cases. In particular, we test the efficacy of this technique by assessing one of the most puzzling cases of recent history, i.e., Aldo Moro’s death. Terrorists of the Red Brigades claimed that they killed Moro when he was placed inside the trunk of a car,shooting him with a barrage of bullets. We demonstrate, based on the analysis of the photographs taken during the autoptic procedure, that the terrorist’s account does not hold true. Our results, showing different series of shots, point towards a three-step execution, with the first phasestaking place outside the car. In conclusion, the novel forensic analysis method introduced in this paper permits the evaluation of a collection of vortex cycles/nerves equipped with a connectedness proximity, which makes it possible to assess unexpected spatial clusters in photographs.
Wed, 5 December 2018
REVIEW Download: 143| View: 207| Comments: 0 | doi:10.20944/preprints201812.0066.v1
Subject: Medicine & Pharmacology, Pathology & Pathobiology Keywords: renal cell carcinoma; circulating DNA; CTC; diagnosis; follow-up; genetic alteration; target therapy
Online: 5 December 2018 (08:01:30 CET)
Liquid biopsy, based on the circulating tumor cells (CTCs) and cell-free nucleic acids has potential applications at multiple points throughout the natural course of cancer, from diagnosis to follow-up. The advantages of doing ctDNA assessment vs. tissue-based genomic profile are the minimal procedural risk, the possibility to serial testing in order to monitor disease-relapse and response to therapy over time and to reduce hospitalization costs during the entire process. However some critical issues related to ctDNA assays should be taken in consideration. The sensitivity of ctDNA assays depends on the assessment technique and genetic platforms used, on tumor-organ, stage, tumor heterogeneity, tumor clonality. The specificity is usually very high, whereas the concordance with tumor-based biopsy is generally low. In patients with renal cell carcinoma (RCC) qualitative analyses of ctDNA have been performed with interesting results regarding selective pressure from therapy, therapeutic resistance, exceptional treatment response to everolimus and mutations associated with aggressive behavior. Quantitative analyses showed variations of cfDNA levels at different tumor stage. Compared to CTC assay, ctDNA is more stable than cells and easier to isolate. Splice variants, information at single-cell level and functional assays along with proteomics, transcriptomics and metabolomics studies can be performed only in CTCs.
Fri, 28 June 2019
ARTICLE Download: 51| View: 171| Comments: 0 | doi:10.20944/preprints201906.0301.v1
Subject: Medicine & Pharmacology, Pathology & Pathobiology Keywords: tumor blood vessel; Tumor Angiogenic Inhibition Triggered Necrosis (TAITN); CXCR4 antagonist; oral squamous cell carcinoma; hypoxia
Online: 28 June 2019 (15:18:43 CEST)
CXCR4 is a chemokine receptor crucial in tumor progression, although the angiogenic role of CXCR4 in oral squamous cell carcinoma (OSCC) has not been investigated. Here we show that CXCR4 is crucial for tumor angiogenesis thereby supports tumor survival in OSCC. Immunohistochemistry on human clinical specimens revealed that CXCR4 and a tumor vasculature marker CD34 were co-distributed in tumor vessels in human OSCC specimens. To ask the effects of CXCR4 inhibition, we treated the OSCC-xenografted mice with AMD3100, so-called plerixafor, an antagonist of CXCR4. Notably, we found a unique pathophysiological structure defined as Tumor Angiogenic Inhibition Triggered Necrosis (TAITN) induced by the CXCR4 antagonism. Treatment with AMD3100 increased necrotic area with the induction of hypoxia-inducible factor-1α in the xenografted tumors, suggesting that AMD3100-induced TAITN was involved in hypoxia and ischemia. Taken together, we demonstrated that CXCR4 plays a crucial role in tumor angiogenesis required for OSCC progression, whereas TAITN induced by CXCR4 antagonism could be an effective anti-angiogenic therapeutic strategy in OSCC treatment.
Wed, 25 July 2018
REVIEW Download: 233| View: 169| Comments: 0 | doi:10.20944/preprints201807.0478.v1
Subject: Medicine & Pharmacology, Pathology & Pathobiology Keywords: pheochromocytoma; paraganglioma; GAPP; metastasis; prognosis; catecholamine; gene mutation; immunohistochemistry; pathology; diagnosis
Online: 25 July 2018 (11:41:52 CEST)
Pheochromocytoma and sympathetic paraganglioma (PPGL) are rare neuroendocrine tumors characterized by catecholamine production in the adrenal medulla and extra-adrenal paraganglia. PPGL with metastasis was termed malignant PPGL. However, the distinction between “benign” and “malignant” PPGLs has been debated. Currently, all PPGLs are believed to have some metastatic potential and are assigned malignant tumors (ICD-O/3) by the WHO Classification of Endocrine Organs (2017, 4th edition). Therefore, the previous categories benign and malignant PPGL have been eliminated in favor of a risk stratification approach. The Grading of Adrenal Pheochromocytoma and Paraganglioma (GAPP) is a tool for risk stratification for predicting metastasis and the prognosis of patients. At least 30% of PPGLs are hereditary, with 20 genes identified and genotype-phenotype correlations clarified. Of these, VHL, RET, and NF1 have been well investigated and are the primary cause of bilateral PCC. In addition, succinate dehydrogenase gene subunits SDHB and SDHD are strongly correlated with extra-adrenal location, younger age, multiple tumors, metastasis, and poor prognosis. Disease stratification by catecholamine phenotype and molecular profiling correlates with histological grading by GAPP. PPGLs should be understood comprehensively based on clinical, biochemical, molecular, and pathological data for patient care. A flow chart for pathological diagnosis is included.
Sat, 28 September 2019
REVIEW Download: 59| View: 165| Comments: 0 | doi:10.20944/preprints201909.0314.v1
Subject: Medicine & Pharmacology, Pathology & Pathobiology Keywords: hypothalamus; endocrine; gangliocytoma; neurocytoma; pituicytoma; hormones
Online: 28 September 2019 (00:24:48 CEST)
The hypothalamus is the site of synthesis and secretion of a number of endocrine peptides that are involved in the regulation of hormonal activity of the pituitary and other endocrine targets. Tumors of the hypothalamus have been recognized to have both structural and functional effects including hormone hypersecretion. The classification of these tumors had advanced over the last few years and biomarkers are now available to classify these tumors and provide accurate structure-function correlations. This review provides an overview of tumors in this region that is critical to metabolic homeostasis with a focus on advances in the diagnosis of gangliocytomas, neurocytomas and pituicytomas that are unique to this region.
Thu, 27 September 2018
REVIEW Download: 225| View: 159| Comments: 0 | doi:10.20944/preprints201809.0533.v1
Subject: Medicine & Pharmacology, Pathology & Pathobiology Keywords: mycoplasma; virulence effectors; pathogenesis
Online: 27 September 2018 (05:54:04 CEST)
Members of the genus Mycoplasma and related organisms impose a substantial burden of infectious diseases on humans and animals, but the last comprehensive review of mycoplasmal pathogenicity was published 20 years ago. Post-genomic analyses have now begun to support the discovery and detailed molecular biological characterization of a number of specific mycoplasmal virulence factors. This review covers three categories of defined mycoplasmal virulence effectors: 1) specific macromolecules including the superantigen MAM, the ADP-ribosylating CARDS toxin, sialidase, cytotoxic nucleases, cell-activating diacylated lipopeptides, and phosphocholine-containing glycoglycerolipids; 2) the small molecule effectors hydrogen peroxide, hydrogen sulfide, and ammonia; and 3) several putative mycoplasmal orthologs of virulence effectors documented in other bacteria. Understanding such effectors and their mechanisms of action at the molecular level connects the biology of the bacteria to direct effects on the host and host responses they elicit, and is expected to translate into new interventions for human and veterinary mycoplasmosis.
Wed, 12 September 2018
ARTICLE Download: 157| View: 147| Comments: 0 | doi:10.20944/preprints201809.0222.v1
Subject: Medicine & Pharmacology, Pathology & Pathobiology Keywords: psoriasis, arthritis, inflammation, granulocytes, redox signaling, oxidative stress, lipid peroxidation, 4-hydroxynonenal, lipids, endocannabinoid system
Online: 12 September 2018 (13:43:16 CEST)
Inflammatory granulocytes are characterized by oxidative burst, which may promote oxidative stress and lipid modification both in affected tissues and on systemic level. On the other hand, redox signaling involving lipid peroxidation products acting as second messengers of free radicals play important, not yet fully understood, roles in pathophysiology of inflammation and various stress-associated disorders. Therefore, the aim of this study was to evaluate the onset of oxidative stress and alterations of enzyme-dependent lipid metabolism resulting from redox imbalance in granulocytes and plasma obtained from patients with psoriasis vulgaris or psoriatic arthritis, in comparison to the healthy subjects. The results obtained revealed enhanced activity of pro-oxidant enzymes NADPH and xanthine oxidases in granulocytes, with a decrease of enzymatic and non-enzymatic antioxidants in plasma of psoriatic patients. The Nrf2 and its regulators were increased in both forms of psoriasis, while HO-1 levels were increased only in psoriasis vulgaris. Redox imbalance was associated with decreased levels of phospholipids and of free PUFAs, but with enhanced activity of enzymes involved in lipid metabolism (PLA2, PAF-AH COX1/2) and increased lipid peroxidation products 4-hydroxynonenal (4-HNE), isoprostanes and neuroprostanes. Increased endocannabinoids and GPR55 were observed in both forms of the disease, while expression of CB1 was increased only in pateints with psoriatic arthritis, opposite to CB2, which was increased only in psoriasis vulgaris. Protein modifications by ROS and lipid peroxidation product 4-HNE promoted apoptosis of granulocytes by increased caspases in both forms of psoriasis. This study indicates that excessive activation of granulocytes, causing oxidative stress and lipid modifications, is an important pathophysiology of psoriasis. Consequently, lower Nrf2 activity and CB2 expression may promote progression of psoriasis into advanced, arthritic form of the disease.
Sun, 28 July 2019
ARTICLE Download: 67| View: 146| Comments: 0 | doi:10.20944/preprints201907.0308.v1
Subject: Medicine & Pharmacology, Pathology & Pathobiology Keywords: orthohantavirus; bunyavirus; VHF; viruses; haemorrhagic fever; epidemiology; infectious diseases; Barbados; Caribbean; virology; incidence; vectors; Americas
Online: 28 July 2019 (11:43:17 CEST)
Analysis of the demographic, temporal and seasonal distribution of hantavirus infections in Barbados was conducted using national surveillance data for 861 laboratory confirmed cases during 2008-2016. The crude incidence rate of hantavirus infections varied from 5.05 to 100.16 per 100,000 persons per year. One major hantavirus epidemic occurred in Barbados during 2010. Hantavirus cases occurred throughout the year with low level transmission during the dry season (December to June) with increased transmission during rainy season (July to November) and a seasonal peak in August. Hantavirus incidence rates were significantly higher in females than males every year during the study period. More than 50% of hantavirus cases were 30 years of age or less. The highest incidence rate (63.36 cases per 100,000 population) was observed among patients 0–4 years of age. This represents the first epidemiological data for hantavirus disease among an entire population in the English-speaking Caribbean.
Mon, 3 December 2018
ARTICLE Download: 129| View: 136| Comments: 0 | doi:10.20944/preprints201812.0017.v1
Subject: Medicine & Pharmacology, Pathology & Pathobiology Keywords: primary brain tumor; HER2; immunohistochemistry
Online: 3 December 2018 (09:24:06 CET)
Background and objectives: Primary brain tumors include any tumors arising in the brain whose prognosis is poor due to their histologic characteristics. The aim of this research was to evaluate the frequency of HER2 tumor marker in primary malignant brain tumors. Materials and Methods: This descriptive study was conducted on the samples admitted to the pathology laboratory with diagnosis of primary brain tumor during 2008–2015. Results: From among 107 patients (61.7% males and the rest females) with mean age of 40.4 years, the highest frequency of tumor location was in supratentorial region of the brain (including lobes and ventricles) (63.85% cases). High-grade astrocytoma had the highest prevalence at diagnosis (43.9%), followed by low-grade astrocytoma (37.4%). As for HER2 score, 42.1% of patients were HER2-positive (scores 2 & 3). On the other hand, 5.6% of patients were HER2-negative (-), 40.2% were positive (+), and 54.2% were positive (++). The patients with high-grade astrocytoma had older age (P < 0.001), higher HER2 positivity (P = 0.024) and percentage (P < 0.001) compared to the patients with low-grade astrocytoma. Conclusions: HER2 expression is dependent on the type of brain tumors. High expression of HER2 in high-grade astrocytoma may be useful for therapeutic purposes. The future research is needed to confirm these results with a large number of patients in different areas.
Tue, 23 July 2019
ARTICLE Download: 72| View: 88| Comments: 0 | doi:10.20944/preprints201907.0246.v1
Subject: Medicine & Pharmacology, Pathology & Pathobiology Keywords: influenza virus; imprinting; haemagglutinin; antibody titre; quadrivalent vaccine; influenza A; H1N1; split-virion; isotype; virus neutralization
Online: 23 July 2019 (04:03:13 CEST)
Influenza virus imprinting is now understood to significantly the influence immune responses and clinical outcome of influenza virus infections that occur later in life. Due to the yearly cycling of influenza viruses, humans are imprinted with the circulating virus of their birth year to subsequently build a complex influenza virus immune history but very little is known about how the imprinting strain influences vaccine responses. To investigate the imprinted host immune responses to split-virion vaccination, we imprinted ferrets with a sublethal dose of the historical seasonal H1N1 strain A/USSR/90/1977. After a +60 day recovery period was given to build immune memory, ferrets were immunized and the challenge at Day 123. Samples were collected throughout the time course and immunological assays were performed to investigate recall mechanisms. The preimmune-vaccinated ferrets did not experience significant disease during challenge while naïve-vaccinated ferrets had severe disease. Hemagglutination inhibition assays showed preimmune ferrets had a more robust antibody response post vaccination, increased virus neutralization activity. Virus specific immunoglobulins were of predominantly the IgG isotype suggesting B cell maturity and plasticity at vaccination. These results are important and should be considered for vaccine design.
Mon, 22 October 2018
CASE REPORT Download: 127| View: 85| Comments: 0 | doi:10.20944/preprints201810.0491.v1
Subject: Medicine & Pharmacology, Pathology & Pathobiology Keywords: cytomegalovirus (CMV); infection, reactivation, epidemiology, host-virus interaction, CMV-specific IgG; protective IgG level, avidity, adoptive/acquired immune response, hematopoietic stem cell transplantation (HSCT); secondary immunodeficiency, Quantiferon, pentamer, β2-microglobulin/beta 2-Microglobulin
Online: 22 October 2018 (11:53:47 CEST)
Although the existing paradigm states that CMV reactivation is under control of cellular immune response, the role of humoral counterpart is underestimated. Anti-CMV positive woman after conditioning with Bu-Flu-ATG underwent stem cell transplantation from fully matched, seronegative sibling donor. In immunodeficient recipient fast and significant decrease of specific immune response was observed but reconstitution of marrow-derived B and NK cells was prior thymic origin T cells. The lowest CMV-IgG(89 U/ml) was observed just before CMV viremia. Noteworthy, the sole and exclusive factor of CMV-specific immune response is a residual recipient antibody class IgG. The CMV-quantiferon increase was observed later, but in the first phase immune reconstitution of the PHA-induced IFNγ release was significantly lower than that CMV-induced. It corresponds with NK cells increase at the top of lymphocyte reconstitution and undetected CMV-specific CD8 cells by pentamer technique. Most of NK cells are CD16+, thus are stimulated by residual IgG. In immunocompetent donor the cellular and humoral immune response increases in parallel manner but symptoms of CMV mononucleosis were observed till the increase of specific IgG. Our observations shed light on a unique host-CMV interaction: indicate that significant decrease of CMV-IgG is a good predictor for CMV reactivation during secondary immunodeficiency.
Sat, 18 January 2020
ARTICLE Download: 23| View: 74| Comments: 0 | doi:10.20944/preprints202001.0195.v1
Subject: Medicine & Pharmacology, Pathology & Pathobiology Keywords: Burkitt's lymphoma; Epstein-Barr Virus
Online: 18 January 2020 (09:01:45 CET)
The Tumor Microenviroment (TME) is a complex milieu that is increasingly recognized as a key factor in multiple stages of disease progression and responses to therapy as well as escape from immune surveillance. However, the precise contribution of specific immune effector and immune suppressor components of the TME in Burkitt lymphoma (BL) remains poorly understood. In this paper, we applied the computational algorithm CIBERSORT to Gene Expression Profile (GEP) datasets of 40 BL samples to draw a map of immune and stromal components of TME. Furthermore, by VECTRA multispectral immunofluorescence (IF) and multiple immunohistochemistry (IHC), we investigated the TME of an additional series of 40 BL cases and evaluated the possible role of the PD-1/PD-L1 immune checkpoint axis. Our results indicated that M2 polarized macrophages are the most prominent TME component in BL. In addition, we investigated the correlation between PD-L1 and latent membrane protein-2A (LMP2A) expression on tumour cells, highlighting a subgroup of BL cases characterized by a non- canonical latency program of EBV with an activated PD-L1 pathway. In conclusion, our study analysed the TME in BL and identified a tolerogenic immune signature highlighting new potential therapeutic targets.
Sat, 4 January 2020
ARTICLE Download: 24| View: 51| Comments: 0 | doi:10.20944/preprints202001.0027.v1
Subject: Medicine & Pharmacology, Pathology & Pathobiology Keywords: cross-tolerance; acute kidney injury; miArray; miR-21a; miR-144-3p; miR-146a-5p; miR-451a; miR-762
Online: 4 January 2020 (04:55:30 CET)
Background: Pre-treatment with lipopolysaccharide (LPS) protected the kidney against a later lethal ischemia. To reveal the mechanisms of renal cross-tolerance and septic acute kidney injury we investigated the effects of LPS on miRNA expression in the kidney. Methods: Male NMRI mice were injected with 40 and 10 mg/kg LPS ip. and sacrificed at 1.5 and 6 hours (early preconditioning, EP) and at 24 and 48 hours (late preconditioning, LP). The miRNA profile was established using miRCURY LNA™ microarray and confirmed with qPCR. Results: Plasma urea concentration peaked at 24 hours after LPS and decreased thereafter. Renal TNF-α and IL-6 mRNA were extremely elevated at all time-points. miRNome changes were mild at 1.5 hours, most miRNAs were altered at 6 and 24 hours and declined by 48 hours. Not all miRNAs could be assayed or validated by qPCR. In EP miR-762 was newly identified and validated and was the most elevated miRNA with both methods. In LP miR-21a-5p was the most influenced miRNA followed by miR-451a, miR-144-3p and miR-146a-5p. MiR-21a-3p increased significantly in both EP and LP. Conclusion: miR-762 might attenuate the LPS-induced immune response during EP and the miR-144/451 cluster is involved in LPS-induced renal preconditioning.