Chapter 13 (Part II): Characterizing Viruses, Viroids, in addition to Prions Viral Detection Methods

Chapter 13 (Part II): Characterizing Viruses, Viroids, in addition to Prions Viral Detection Methods

Chapter 13 (Part II): Characterizing Viruses, Viroids, in addition to Prions Viral Detection Methods

Connell, Jeff, Midday On-Air Talent has reference to this Academic Journal, PHwiki organized this Journal Chapter 13 (Part II): Characterizing Viruses, Viroids, in addition to Prions Detection of Viruses Electron microscopy Immunologic Assays: Detect specific viral proteins or antibodies to them. ELISA (Enzyme Linked Immuno Sorbent Assay) Western Blotting: Detects viral proteins Biological Assays: Detect cytopathic effects (CPE) caused by viral infection of cells. Plaque assays as long as lytic viruses Focus as long as mation as long as trans as long as ming oncogenic viruses Hemagglutination Assay: Many viruses clump red blood cells. Molecular Assays: Assay as long as viral nucleic acids. PCR (Polymerase chain reaction) Southerns (DNA) or Northerns (RNA) Viral Detection Methods ELISA Test as long as Antibodies Plaque Assays Hemagglutination

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Antiviral Therapeutic Agents Agent Virus Mechanism Amantadine Influenza Inhibits uncoating Oseltamivir Influenza Inhibits attachment in addition to (Tamiflu) release from cells Acyclovir Herpes viruses Inhibits DNA polymerase Gancyclovir Cytomegalovirus Inhibits DNA polymerase Ribivarin Respiratory syncitial Inhibits viral enzymes as long as Hepatitis A, measles guanine biosynthesis Azidothymidine HIV Inhibits reverse transcriptase Protease Inhibitors HIV Block viral proteases Interferon Cytomegalovirus Inhibits protein synthesis Hepatitis B Degrades ssRNA VIRAL VACCINES First vaccine was used by Jenner (1798) against smallpox in addition to contained live vaccinia (cowpox) virus. I. Live attenuated vaccines: Mutant viral strains produce an asymptomatic infection in host. Examples: Polio (oral, Sabin vaccine), measles, yellow fever, mumps, rubella, chickenpox, in addition to Flu-Mist (influenza). Advantages: Better immune response Disadvantages: May cause disease due to contamination, genetic instability, or residual virulence. Oral polio vaccine recently discontinued in U.S. II. Killed or inactivated vaccines: Virus is typically grown in eggs or cell culture in addition to inactivated with as long as malin. Examples: Polio (shots, Salk vaccine), rabies, in addition to influenza A & B. Advantages: Immunization with little or no risk of infection. Disadvantages: Less effective immune response, inactivation may alter viral antigens. Toxic preservatives ( as long as malin, thimerasol, etc.) III. Recombinant vaccines: Viral subunits are produced by genetically engineered cells. Example: Hepatitis B Advantages: Little or no risk of infection. Disadvantages: Less effective immune response.

Flu Vaccine is Made from Eggs History of Vaccines 1798: Smallpox vaccine results published by Jenner 1885: Rabies vaccine developed by Pasteur 1906: Pertussis (whooping cough) vaccine developed 1928: Diphtheria vaccine developed 1933: Tetanus toxoid vaccine developed 1946: DPT combination vaccine becomes available 1955: Polio inactivated vaccine (IPV) licensed by Salk 1963: Polio oral vaccine (OPV) developed by Sabin 1963: Measles vaccine developed 1968: Mumps vaccine developed 1969: Rubella/German measles vaccine developed 1972: U.S. ended routine smallpox vaccination 1978: Pneumococcal vaccine becomes available 1979: MMR combination vaccine added to routine childhood immunization schedule History of Vaccines 1987: Hemophilus influenzae type B (Hib) vaccine licensed 1988: Vaccine Injury Compensation Program funded 1991: Hepatitis B recombinant vaccine recommended as long as infants. Vaccine was licensed in 1986. 1995: Varicella (chickenpox) vaccine licensed 1996: DTaP (acellular Perstussis) vaccine licensed as long as children under 18 mo.; believed to be safer than DTP. 1998: Rotavirus vaccine licensed as long as diarrheal disease 1999: Rotavirus vaccine removed as long as safety reasons 2000: Polio oral vaccine removed as long as safety reasons Prevnar (Pneumococcal conjugate vaccine) licensed 2002: Thimerasol use as vaccine preservative in most pediatric vaccines discontinued as long as safety reasons 2002: Flumist (inhaled flu vaccine) reviewed by FDA 2007: Gardasil (HPV) in addition to Menactra (meningitis) vaccines licensed

Vaccine Safety Concerns Adverse Reactions: May occur almost immediately or within days, weeks, or months of vaccination. 1. Toxic Effects: Bacterial Toxins: Killed bacterial vaccines can release toxins into the bloodstream. May be associated with swelling, soreness, fever, behavioral in addition to neurological problems (ADHD, autism, etc.). Vaccine Ingredients: May cause neurological, immunological, digestive, or other problems. Thimerosol is a preservative used as long as multiple dose vaccines that contains 49% ethylmercury. Removed from most pediatirc vaccines in 2002. Other ingredients: Aluminum, as long as maldehyde, benzethonium chloride, ethylene glycol, glutamate, phenol, etc. Vaccine Safety Concerns 2. Immune Reactions: Autoimmune: Patient makes antibodies that cross react with host antigens. May cause rheumatoid arthritis, juvenile diabetes, multiple sclerosis, Crohn’s disease (bowel inflammation), Guillain-Barre syndrome (muscle weakness, paralysis), in addition to encephalitis. Suspect vaccines include measles, tetanus, in addition to influenza shots. Allergic reactions: Vaccine ingredients may induce allergic reactions in addition to /or anaphylactic shock in certain individuals. E.g.: Eggs, gelatin, neomycin, in addition to streptomycin. 3. Infectious Viruses: Live attenuated virus vaccines can mutate back to a harmful as long as m in addition to cause the disease they are designed to prevent: oral polio, measles, mumps, rubella, in addition to chickenpox vaccines. Vaccines may be contaminated with other viruses. Vaccine Safety Concerns Can Vaccines Cause Autism Modern Epidemic: A 2009 report in Journal of Pediatrics indicates that one in 100 children in the United States are autistic. In 1960s incidence was 1 in 2,000. Boys are more heavily affected than girls (about 1 in 50 boys; 4-5 X higher rates of autism). Onset of symptoms usually occurs between 1 in addition to 3 years of age. Symptoms in Three Areas: Can vary from severe to mild. Communication: Loss of language, language delays, poor expressive in addition to receptive language Repetitive behaviors: Stimming, h in addition to flapping, running in circles, rocking, obsessions with certain things Social Difficulties: Poor eye contact, isolation, aggression, lack of social skills, tantrums Cause: Unknown. Traditional treatment focuses on symptoms: speech, occupational in addition to behavioral therapy.

Vaccine Safety Concerns Can Vaccines Cause Autism Biomedical Hypothesis: Multiple vaccines in addition to toxins at early age overwhelm immune system. May cause persistent infections. Impaired immunity: Frequent ear infections, colds, etc. Repeated use of antibiotics to treat infections may wipe out beneficial microbial flora in addition to allow “bad microbes” (yeasts in addition to others) to overgrow (gut dysbiosis) Digestive problems: Inability to digest in addition to absorb certain foods (milk, gluten, in addition to others). May develop multiple food intolerances in addition to allergies. Neurological symptoms: May be caused by gut dysbiosis (microbial toxins) in addition to digestive problems (caseomorphin, gliadorphin). Alternative biomedical therapies: Special diet (Gluten free/Casein free), probiotics, antifungals, in addition to nutritional supplements. Use of revised vaccine schedule. Hepatitis B is a Major Health Threat Fifth leading cause of deaths due to infectious disease in the world. 2 million deaths per year. Over 300 million infected individuals worldwide. In Southeast Asia in addition to Africa 10% population is infected. In North America in addition to Europe 1% population is infected. Highly contagious. Virus particles found in saliva, blood, in addition to semen. Mechanisms of transmission: Mother to infant: Primarily at birth. Intimate or sexual contact. Blood transfusions or blood products. Direct contact with infected individuals: Health care workers. Infectious Diseases Causing Most Deaths Worldwide in 2000 Disease Cause Deaths/year Acute Respiratory Bacterial or viral 4,400,000 Diarrheal diseases Bacterial or viral 3,200,000 Tuberculosis Bacterial 3,100,000 Malaria Protozoan 3,100,000 Hepatitis B Viral 2,000,000 Measles Viral 1,500,000 AIDS Viral 1,000,000 Neonatal Tetanus Bacterial 600,000 : Pneumonia, bronchitis, influenza, etc.

Characteristics of Hepatitis B Infection Incubation period: 2 to 6 months. Several possible outcomes: Asymptomatic infection: Most individuals. Acute Hepatitis: Liver damage, abdominal pain, jaundice, etc. Strong immune response usually leads to a complete recovery. Fulminant Hepatitis: Usually fatal. Rare. Chronic Hepatitis: Poor immune response to virus, which remains active as long as years. May be healthy, experience fatigue, or have persistent hepatitis. Associated with development of liver cancer in addition to cirrhosis. Characteristics of Hepatitis B Infection Several possible outcomes: Cirrhosis of the Liver: Severe organ damage, leading to liver failure. Hepatocellular Carcinoma: Usually takes 30 to 50 years to develop. May develop in children. Herpesviruses Family of over 100 viruses which infect a broad range of animals. Polyhedral capsid: Icosahedral capsid, 100-110 nm in diameter. Envelope: Contains viral glycoproteins on its surface. Virion is about 200 nm in diameter. Tegument: Unique to herpesviruses. Amorphous material surrounding capsid. Contains several viral proteins. Large genome: 140-225 kb of linear dsDNA which circularizes after infection.

Biological Properties of Herpesviruses Encode large array of enzymes involved in nucleic acid metabolism. Synthesis of viral DNA in addition to assembly of capsid occurs in the nucleus. Production of infectious progeny causes destruction of infected cell. Latency: Can remain latent in their natural hosts. Viral DNA remains as closed circular molecule in addition to only a few viral genes are expressed. Establish life-long infections. Human Herpesviruses Virus Common Name/Disease Class Size Latency HHV-1 Herpes simplex 1 (HSV-1) a 150 kb Sensory Oral, ocular lesions, encephalitis nerve ganglia HHV-2 Herpes simplex 2 (HSV-2) a 150 kb Sensory Genital lesions, neonatal infections nerve ganglia HHV-3 Varicella zoster virus a 130 kb Sensory Chickenpox, shingles nerve ganglia HHV-4 Epstein-Barr virus g 170 kb B cells Mononucleosis, tumors` Salivary gl in addition to HHV-5 Human Cytomegalovirus b 230 kb Lymphocytes Microcephaly, infections in immunocompromised hosts HHV-6/7 Human Herpesvirus 6/7 b 160 kb CD4 T cells Roseola Infantum HHV-8 Human Herpesvirus 8 g 140 kb Kaposi’s Kaposi’s sarcoma, lymphoma Sarcoma tissue Clinical Manifestations of HSV-1 Epidemiology: 70-90% of adults are infected. Most are asymptomatic. Gingivostomatitis: Most common manifestation of primary HSV-1 infection. Initial infection typically occurs in early childhood. Recurrent herpes labialis: Cold sores, fever blisters. After primary disease, virus remains latent in trigeminal ganglion. During reactivation, virus travels down nerve to peripheral location to cause recurrence. Whitlow: Infection of finger.

Recurrent Herpes Labialis Less than 1 day with erythema in addition to burning Same patient 24 h later with multiple fluid filled vesicles in addition to erythema Recurrent Herpes Labialis: Bilateral vesicles on upper in addition to lower lips. Source: Atlas of Clinical Oral Pathology, 1999. Herpetic Whitlow: Multiple crusting ulcerations that begin as vesicles. Source: Atlas of Clinical Oral Pathology, 1999.

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Keratoconjunctivitis: Most common cause of corneal blindness in US. Eczema herpeticum: Severe herpetic outbreaks in areas with eczema. Herpes gladiatorum: Inoculation of abraded skin by contact with infected secretions. HSV encephalitis: Most common cause of acute sporadic encephalitis in US. Chronic herpes simplex infection: Lesions in atypical oral locations. Immunocompromised patients. Chronic Herpes Simplex infection with lesions on tongue in addition to lips. Source: Atlas of Clinical Oral Pathology, 1999. Clinical Manifestations of HSV-2 Epidemiology: Acquisition follows typical pattern of STD. Seroprevalence ranges from 10% to 80% Most individuals are asymptomatic. Genital Herpes: Most common manifestation HSV-2 infection. Most common cause of genital ulcers in U.S. Lesions on cervix, perineum, or penis shaft. Recurrence rates vary widely. Perirectal Herpes: Can be severe in AIDS patients. Orofacial herpes: Less than 5% of cases. Neonatal Herpes: Due to contact with infected genital secretions during delivery. Severe disease with encephalitis, pneumonitis, hepatitis, in addition to retinitis.

Herpes simplex 2 infection with fluid filled vesicles on penis. Source: Mike Remington, University of Washington Viral Disease Clinic Genital Herpes Acyclovir resistant peri-rectal HSV2 infection in HIV infected male. Source: AIDS, 1997 Clinical Manifestations of Varicella Zoster (HHV-3) Chickenpox (Varicella): Most common manifestation of primary herpes zoster infection. Epidemiology: Highly communicable. Airborne or skin transmission. Incubation period 14 days. Be as long as e the vaccine (Varivax) was introduced in 1995, there were about 3 million cases/year in US (most in the spring). Since 1995, the number of cases has dropped by 85%. Symptoms: Malaise, sore throat, rhinitis, in addition to generalized rash that progresses from macules to vesicles. Intraoral lesions may precede rash. Complications: Reye’s syndrome, bacterial superinfection of lesions, varicella pneumonia in addition to neonatal varicella (30% mortality).

Prions First described by Stanley Prusiner in 1982. Won Nobel Prize in 1997 Infectious proteins called PrP that lack nucleic acids All mammals contain a gene that codes as long as the PrP protein Two as long as ms of PrP with different tertiary structures: Normal Cellular Prp (alpha-helices) Prion PrP (Beta pleated sheets) Cause nine known neurological diseases called transmissible spongi as long as m encephalopathies (TSEs) No known treatment. Currently 100% fatal Prion Proteins cause Transmissible Spongi as long as m Encephalopathies in Animals Known Transmissible Spongi as long as m Encephalopathies (TSEs) Cows: Mad cow disease or Bovine Spongi as long as m Encephalitis (1987 outbreak in UK). Linked to feeding scrapie infected sheep to cattle Human diseases: Tendency to run in families. Creutzfeldt-Jakob Disease (CJD): 200 cases/year in US, genetic, surgery, transplants Variant CJD: Linked to eating infected beef (150+ cases worldwide) Kuru: Associated with cannibalism Gertsmann-Straussler-Scheinker Syndrome Fatal Familial Insomnia Sheep: Scrapie outbreak in 1940s in U.S. Deer in addition to elk: Chronic Wasting Disease Mink: Transmissible Mink Encephalopathy

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