Lebanese Plants and Plant-Derived Compounds Against Colon Cancer

Colorectal cancer (CRC) is a major health concern and demands long-term efforts in developing strategies for screening and prevention. CRC has become a preventable disease as a consequence of a better understanding of colorectal carcinogenesis. However, current therapy is unsatisfactory and necessitates the exploration of other approaches for the prevention and treatment of cancer. Plant based products have been recognized as preventive with regard to the development of colon cancer. Therefore, the potential chemopreventive use and mechanism of action of Lebanese natural product were evaluated. Towards this aim the antitumor activity of Onopordum cynarocephalum and Centaurea ainetensis has been studied using in vitro and in vivo models. In vitro, both crude extracts were non cytotoxic to normal intestinal cells and inhibited the proliferation of colon cancer cells in a dose-dependent manner. In vivo, both crude extracts reduced the number of tumors by an average of 65% at weeks 20 (adenomas stage) and 30 (adenocarcinomas stage). The activity of the C. ainetensis extract was attributed to Salograviolide A, a guaianolide-type sesquiterpene lactone, which was isolated and identified through bio-guided fractionation. The mechanism of action of thymoquinone (TQ), the active component of Nigella sativa, was established in colon cancer cells using in vitro models. By the use of N-acetyl cysteine, a radical scavenger, the direct involvement of reactive oxygen species in TQ-induced apoptotic cells was established. The analytical detection of TQ from spiked serum and its protein binding were evaluated. The average recovery of TQ from spiked serum subjected to several extraction procedures was 2.5% proving the inability of conventional methods to analyze TQ from serum. This has been explained by the extensive binding (>98%) of TQ to serum and major serum components such as bovine serum albumin (BSA) and alpha-1-acid glycoprotein (AGP). Using mass spectrometry analysis, TQ was confirmed to bind covalently to the free cysteine in position 34 and 147 of the amino acid sequence of BSA and AGP, respectively. The results of this work put at the disposal for future development new plants with anti-cancer activities and enhance the understanding of the pharmaceutical properties of TQ, a prerequisite for its future clinical development.Paksusuolen syöpä on merkittävä terveyttä uhkaava sairaus ja vaatii pitkäjänteisiä ponnisteluita kehitettäessä lähestymistapoja taudin toteamiseksi ja hoitamiseksi. Paksusuolen syövän hoito on parantunut ja paksusuolen syövän kehittymiseen vaikuttavat tekijät tunnetaan paremmin kuin aikaisemmin. Kuitenkin nykyiset hoitokäytännöt eivät ole täysin tyydyttäviä ja tekevät välttämättömäksi, että tutkitaan muita tapoja ennalta ehkäistä ja hoitaa syöpää. On havaittu kasviperäisten tuotteiden estävän paksusuolen syövän kehittymistä. Tämän johdosta tässä tutkimuksessa libanonilaisten luonnontuotteiden syöpää estävää vaikutusta ja vaikutusmekanismia tutkittiin. Tutkittiin Onopordum cynarocephalum ja Centaurea ainetensis kasvien vaikutuksia paksusuolen syöpään käyttämällä in vitro and in vivo solu- ja eläinkoe malleja. In vitro, solukokeissa molemmat raakauutteet eivät olleet sytotoksisia suolistosoluille ja estivät paksusuolen syöpäsolujen lisääntymistä annosvasteisesti. In vivo, eläinkokeissa molemmat raakauutteet vähensivät kasvaimien määrää keskimäärin 65%:lla 20 viikkoa hoidon aloittamisesta (adenooma vaihe) and 30 viikkoa hoidon aloittamisesta (adenokarsinooma vaihe). C. ainetensis-uutteen aktiivisuus voitiin liittää Salograviolidi A:han, guaianolidi-tyyppinen seskviterpeenilaktoniin, mikä eristettiin ja tunnistettiin aktiivisuuden ohjaaman eristyksen avulla. Nigella sativa-kasvin vaikuttavan aineen tymokinonin (TQ) vaikutusmekanismia tutkittiin käyttämällä paksusuolen syöpäsolumalleja in vitro. Käyttämälllä N-asetyylikysteiiniä, radikaalin sieppaajaa, reaktiivisen happimolekyylien suora osallistuminen TQ-indusoiduissa apoptoottisissa soluissa voitiin osoittaa. TQ:n mittaamista seerumista ja TQ:n sitoutumista proteiineihin tutkittiin. Kun TQ oli lisätty seerumiin sen keskimääräinen saanto useiden uuttojen jälkeen oli vain 2.5%, mikä osoitti tavanomaisten menetelmien heikkouden analysoitaessa TQ:ta seerumista. Tämä selittyi TQ:n voimakkaalla sitoutumisella (>98%) seerumiin ja seerumin pääkomponentteihin kuten naudan seerumin albumiini (BSA) ja hapan glykoproteiini (alpha-1-acid glycoprotein, AGP). Käyttämällä massaspektrometri-analytiikkaa TQ:n voitiin osoittaa sitoutuvan kovalenttisesti vapaaseen kysteiiniin aminohappoissa 34 (BSA) ja 147 (AGP). Tämän työn tulokset mahdollistavat tulevaisuudessa uusien kasviperäisten syöpälääkkeiden kehittämisen ja edesauttavat TQ:n farmaseuttisten ominaisuuksien ymmärtämistä kehitettäessä sitä tulevaisuudessa lääkkeeksi kliiniseen käyttöön

LATE EVENTS OF HUMAN METAPNEUMOVIRUS INFECTION: INSIGHTS INTO VIRAL SPREAD WITHIN THE RESPIRATORY EPITHELIUM

Human metapneumovirus (HMPV) is a leading cause of respiratory tract infections worldwide across all age groups, and is particularly devastating in the pediatric, elderly and immunocompromised populations. Despite its high prevalence and burden on human health, there are currently no treatments or vaccines against HMPV infections. HMPV is an enveloped virus that belongs to the paramyxovirus family. Paramyxoviruses in general form by assembly of virus components at the plasma membrane followed by budding and release of virus particles into the extracellular matrix to spread infection. The process of forming new virus particles requires complex interactions between viral and cellular components and the requirements for particle production differ substantially among paramyxoviruses. Several key aspects of the life cycle of HMPV remain unknown. The work presented here provides significant advances in understanding the mechanisms underlying assembly and spread of HMPV in human bronchial airway epithelial cells. We provide evidence that HMPV induces reorganization of the actin cytoskeleton and microtubules at late stages of infection leading to the formation of complex networks of branched filaments and intercellular extensions, structures that have not been previously reported for paramyxoviruses. Our results indicate a novel mode of HMPV spread directly from cell-to-cell across intercellular extensions. We identified an important role of actin and the Rho GTPases Rac1 and Cdc42 in direct cell-to-cell spread of HMPV. While roles for paramyxovirus matrix and fusion proteins in membrane deformation have been previously demonstrated for several paramyxoviruses, we show that the HMPV phosphoprotein (P) associates with actin and induces formation of membrane extensions, suggesting a role for the P protein in HMPV exit from the cell. Additionally, infection of differentiated, polarized human airway tissues showed that release of HMPV particles at the apical side is inefficient and revealed that spread of HMPV in these tissues can occur in the presence of neutralizing antibodies. HMPV infection also resulted in reorganization of the actin cytoskeleton in these tissues mainly at the apical side. Collectively, the data provided in this dissertation reveal a novel mechanism by which HMPV uses the cytoskeleton for cell-to-cell transmission and provide critical insights into spread of respiratory viruses within the airway epithelium

DNA translocases and origin region segregation in B. subtilis

Specialized mechanisms involved in chromosome segregation, septum placement, and chromosome dimer resolution contribute to the maintenance of chromosome integrity throughout the cell cycle. The first part of this work focuses on the investigation of DNA translocases in the Gram positive model organism Bacillus subtilis, which move the chromosomes away from the division plane by directed DNA transport. SpoIIIE is a membrane-integral translocase that also acts during sporulation, while SftA is associated with the division septum by an unknown mechanism and was reported to be a soluble protein in vitro. The solubility of SftA in vivo was proven with cell fractionation experiments, and the part of the protein that serves for septal targeting was determined through the localization of different truncations of fluorescently labeled SftA, which helped narrow down the stretch of amino acids responsible for targeting the protein to the septum. Expression in a Eukaryotic heterologous system revealed an interaction between SftA and FtsA, but not FtsZ. Further evidence was provided by single molecule tracking experiments whereby the fraction of soluble SftA molecules increased in an FtsA depletion background as compared to a wild type background. Concomitant investigation on the single molecule level of SpoIIIE, a membrane associated DNA translocase in B. subtilis, and PfkA, a soluble phosphofructokinase, revealed a different behavior of the two translocases (SftA and SpoIIIE): SftA has a septal bound fraction, and a small soluble fast moving fraction,comparable in diffusion coefficient to the exclusively soluble PfkA tracked under the same conditions. SpoIIIE is much slower in comparison, and even its “fast” moving fraction is much slower than that of SftA. It seems to move slowly along the membrane with no specific enrichment at the septum, even after Mitomycin C (MMC) treatment. The second part of this work focused on the count of nucleoids, origin, and terminus of replication regions under conditions of fast and slow growth, with or without the induction of double strand breaks with MMC. B. subtilis seemed to be predominantly diploid, with a fraction of polyploid and monoploid cells which changed depending on the growth conditions. Replication of the origin proximal regions increased after DNA damage induction, as was shown by the increase in the number of origins during the time of DNA repair, while the number of termini remained constant. Time lapse experiments of the segregation of the tagged origin regions revealed that the movement is best described as directed diffusion, but seems to be quite robust and continues, though slower, after MMC treatment or treatment with Ciprofloxacin which blocks the topoisomerase I

Patterns of Prehistoric Epidemiology and Human Paleopathology

Human paleopathologists are interested in the visible marks of diagnosable disease that reflect various aspects of human biocultural interaction. Whether infectious, nutritional, or a combination of both, pathological characteristics in the dry bone provide some insight into the health of past human populations. Paleoepidemiology and human paleopathology are important parts of ecology in that they deal directly with a major aspect of man\u27s relationship to his environment. The significance of this relationship has, to a large extent, been neglected by human skeletal biologists. The purpose of this study is to examine one of the most important aspects of human biocultural interaction: patterns of nutritional stress

Variables Associated with Divorce in Northwest Georgia, 2007-2016

Variables Associated with Divorce in Northwest Georgia, 2007-2016 Abstract In this paper, there is an investigation of the main variables associated with divorce in Northwest Georgia, from data collected through 191 interviews, in 2007-2016. The data showed that sixteen such variables were identified by divorced interviewees. The study showed that marriage dissatisfaction and lack of readiness for marriage are the main factors, which explain divorce. The most frequently-mentioned variables were those related to infidelity, communication, abuse, financial problems, and conflict resolution

Online Teaching in Sociology: Prospects, Successes, and Problems

Online teaching is promising to expand more in the future, as it meets student demands and provides more profit to the institutions of higher education offering it. However, it has two main challenges, represented by the physical absence of instructors and the perception that it’s easier to pass online courses than face-to-face courses. Using my ten-year experience in online teaching, I describe my best practices, offering solutions to these challenges and other problems. To engage students, I use a variety of course activities, such as chapter online tests, papers, structured discussion posts, substantive responses to classmates, as well as timely and frequent communication with students. The accusation leveled against online courses as easier to pass than face-to-face courses can be refuted by requiring two proctored unit exams, with considerable weight, as credible measures of the student learning of the course content

Comparison of Three Levels of Divorce in Whitfield County, State of Georgia, and the United States in Two Decades, 1990-2009

Comparison of Three Levels of Divorce in Whitfield County, State of Georgia, and the United States in Two Decades, 1990-2009 Abstract: In this paper, there is an investigation of the North West Georgia divorce ratios in 1990-2010 and how they are related to the same ratios in the State of Georgia and the United States as a whole. The data showed that there was a decline in divorce ratios on the three levels. However, the North West Georgia divorce ratios were still much higher than the State and the national ratios. I would like to thank the three anonymous reviewers for their valuable suggestions

Introduction to Sociology (Dalton State College)

This Grants Collection for Introduction to Sociology was created under a Round Three ALG Textbook Transformation Grant. Affordable Learning Georgia Grants Collections are intended to provide faculty with the frameworks to quickly implement or revise the same materials as a Textbook Transformation Grants team, along with the aims and lessons learned from project teams during the implementation process. Documents are in .pdf format, with a separate .docx (Word) version available for download. Each collection contains the following materials: Linked Syllabus Initial Proposal Final Reporthttps://oer.galileo.usg.edu/psychology-collections/1005/thumbnail.jp

Adverse Effects of Botulinum Toxin type-A injections in masticatory muscles on underlying bone and cartilage: a literature review

Botulinum toxin type A (BTA) injections in masticatory muscles are used to treat numerous clinical conditions. This neurotoxin causes atrophy and transient paralysis of the concerned muscles.The aim of this review is to gather and define the effects of BTA injections in masticatory muscles on the underlying cartilage and bone structures.Electronic search of Medline and Google scholar databases covering the period between January 2007 and July 2019 was carried out. Eligible articles were selected according to the inclusion/exclusion criteria. Fourteen articles were included. BTA injections may cause short term reduction of cortical bone thickness and trabeculation. On the long term, cartilage volume reduction and bone volume loss were evident on the condyle and mandibular angle, at bothinjected and non-injected mandibular bone structures. Bone mineral density remained unaffected when only one masticatory muscle was injected but was reduced when several masticatory muscles were treated with BTA. Several cellular and molecular alterations were noticed in some articles.Because of evidence of irreversible negative effects of BTA on the underlying structures, possible muscle, bone and cartilage volume reduction should be communicated to the patients prior to any intervention. Further studies are needed to fully understand the cellular mechanisms and molecular responses behind this phenomenon

Metal Additive Manufacturing for Fixed Dental Prostheses

The use of additive manufacturing (AM) in dentistry has gained momentum in recent years. However, high initial costs and uncertainty surrounding the quality of AM products are considered barriers to their use. This research compared dental substructures fabricated by AM versus conventional casting and milling. Cobalt-chromium alloy rectangular bars and three-unit bridge substructures were fabricated by AM, casting or milling. Bars manufactured by AM exhibited superior flexural strength, shear bond strength of porcelain coating, and Vickers hardness. Bridge substructures fabricated by AM showed similar flexural stiffness to cast, similar flexural loads at failure to milled and cast, and overall accuracy of fabrication within 12 micrometers. Cast substructures showed the greatest internal porosity, while samples fabricated by AM exhibited pores primarily within the abutment region. Overall, bars and three-unit bridges manufactured by AM exhibited equal or better mechanical properties than those fabricated by conventional techniques