DR. SAMIR CHACHOUA M.B.B.S.

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PRESENTATION BY DR. SAMIR CHACHOUA M.B.B.S.
AVIAN INFLUENZA JANUARY 2006
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Virulent Mutation Ebola Virus Mahrburg
Virus Avian Flu
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Introduction Rapidly mutating viruses pose
particular difficulties for the human organism as
well as in the development of coping strategies.
Vaccines have met with very limited success in
dealing with these diseases. The seasonal
fluctuation of the influenza virus, for example,
proceeds unhindered despite massive campaign of
flu vaccinations
4
AVIAN FLU
H5N1 is a subtype of the species called avian
influenza virus (bird flu). Avian flu is a
disease and avian flu virus is a species. Flu
viruses are named for their Hemagglutinin (H) and
Neuraminidase(N) genes. Avian flu viral subtype
is H5N1.
Influenza A virus, the virus that causes Avian
flu. Transmission electron micrograph of
negatively stained virus particles in late
passage. (Source Dr. Erskine Palmer, Centers for
Disease Control and Prevention Public Health
Image Library).
Colorized transmission electron micrograph of
H5N1 (golden) grown in Madin-Darby canine kidney
cells (green). (Source C. Goldsmith, J. Katz and
S. Zaki. Centers for Disease Control Prevention
Public Health Image Library. Image 1841.).
5
MAGNITUDE EXTENT OF THE PROBLEM
Outbreak of human cases from birds in Hong Kong
in 1997. Current epidemic since 2003. As of
January 11, 2006, 155 cases of infections in
humans, resulting in 78 deaths, have been
confirmed worldwide. Not all cases of H5N1
infection are reported and consequently the exact
mortality rate is unknown. Tens of millions of
birds died of H5N1 influenza and hundreds of
millions of birds were culled (slaughtered and
disposed of) to protect humans from
H5N1. Thirteen countries across Asia and Europe
have been affected. Infected wild birds found in
Europe.
Bird Flu Spread on October 26th, 2005. Official
UN FAO
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MAGNITUDE EXTENT OF THE PROBLEM BY COUNTRY
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PATHOGENESIS OF AVIAN INFLUENZA VIRUS
Characterization of Virus Studies of isolates of
avian influenza A (H5N1) from patients in 1997
revealed that virulence factors included the
highly cleavable hemagglutinin that can be
activated by multiple cellular proteases, a
specific substitution in the polymerase basic
protein 2 (Glu627Lys) that enhances replication,
and a substitution in nonstructural protein 1
(Asp92Glu) that confers increased resistance to
inhibition by interferons and tumor necrosis
factor (TNF-) in vitro and prolonged replication
in swine, as well as greater elaboration of
cytokines, particularly TNF-, in human
macrophages exposed to the virus. Since 1997,
studies of influenza A (H5N1) indicate that these
viruses continue to evolve, with changes in
antigenicity and internal gene constellations an
expanded host range in avian species and the
ability to infect felids enhanced pathogenicity
in experimentally infected mice and ferrets, in
which they cause systemic infections and
increased environmental stability. Phylogenetic
analyses indicate that the Z genotype has become
dominant and that the virus has evolved into two
distinct clades, one encompassing isolates from
Cambodia, Laos, Malaysia, Thailand, and Vietnam
and the other isolates from China, Indonesia,
Japan, and South Korea Recently, a separate
cluster of isolates has appeared in northern
Vietnam and Thailand, which includes variable
changes near the receptor-binding site and one
fewer arginine residue in the polybasic cleavage
site of the hemagglutinin. However, the
importance of these genetic and biologic changes
with respect to human epidemiology or virulence
is uncertain.
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PATHOGENESIS OF AVIAN INFLUENZA VIRUS
Patterns of Viral Replication The virologic
course of human influenza A (H5N1) is
incompletely characterized, but studies of
hospitalized patients indicate that viral
replication is prolonged. In 1997, virus could be
detected in nasopharyngeal isolates for a median
of 6.5 days (range, 1 to 16), and in Thailand,
the interval from the onset of illness to the
first positive culture ranged from 3 to 16 days.
Nasopharyngeal replication is less than in human
influenza,27and studies of lower respiratory
tract replication are needed. The majority of
fecal samples tested have been positive for viral
RNA (seven of nine), whereas urine samples were
negative. The high frequency of diarrhea among
affected patients and the detection of viral RNA
in fecal samples, including infectious virus in
one case, suggest that the virus replicates in
the gastrointestinal tract. The findings in one
autopsy confirmed this observation. Highly
pathogenic influenza A (H5N1) viruses possess the
polybasic amino acid sequence at the
hemagglutinin-cleavage site that is associated
with visceral dissemination in avian species.
Invasive infection has been documented in
mammals, and in humans, six of six serum
specimens were positive for viral RNA four to
nine days after the onset of illness. Infectious
virus and RNA were detected in blood,
cerebrospinal fluid, and feces in one patient.
Whether feces or blood serves to transmit
infection under some circumstances is unknown.
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PATHOGENESIS OF AVIAN INFLUENZA VIRUS
Host Immune Responses The relatively low
frequencies of influenza A (H5N1) illness in
humans despite widespread exposure to infected
poultry indicate that the species barrier to
acquisition of this avian virus is substantial.
Clusters of cases in family members may be caused
by common exposures, although the genetic factors
that may affect a host's susceptibility to
disease warrant study. The innate immune
responses to influenza A (H5N1) may contribute to
disease pathogenesis. In the 1997 outbreaks,
elevated blood levels of interleukin-6, TNF-,
interferon-, and soluble interleukin-2 receptor
were observed in individual patients, and in the
patients in 2003, elevated levels of the
chemokines interferon-inducible protein 10,
monocyte chemoattractant protein 1, and monokine
induced by interferon- were found three to eight
days after the onset of illness.27 Recently,
plasma levels of inflammatory mediators
(interleukin-6, interleukin-8, interleukin-1, and
monocyte chemoattractant protein 1) were found to
be higher among patients who died than among
those who survived (Simmons C personal
communication), and the average levels of plasma
interferon- were about three times as high among
patients with avian influenza A who died as among
healthy controls. Such responses may be
responsible in part for the sepsis syndrome,
ARDS, and multiorgan failure observed in many
patients. Among survivors, specific humoral
immune responses to influenza A (H5N1) are
detectable by microneutralization assay 10 to 14
days after the onset of illness. Corticosteroid
use may delay or blunt these responses.
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PATHOGENESIS OF AVIAN INFLUENZA VIRUS
Pathological Findings Limited postmortem
analyses have documented severe pulmonary injury
with histopathological changes of diffuse
alveolar damage, consistent with findings in
other reports of pneumonia due to human influenza
virus. Changes include filling of the alveolar
spaces with fibrinous exudates and red cells,
hyaline-membrane formation, vascular congestion,
infiltration of lymphocytes into the interstitial
areas, and the proliferation of reactive
fibroblasts. Infection of type II pneumocytes
occurs. Antemortem biopsy of bone marrow
specimens has shown reactive histiocytosis with
hemophagocytosis in several patients, and
lymphoid depletion and atypical lymphocytes have
been noted in spleen and lymphoid tissues at
autopsy. Centrilobular hepatic necrosis and acute
tubular necrosis have been noted in several
instances.
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MODE OF INFECTION OF AVIAN INFLUENZA VIRUS
H5N1 RK7First sampled in Qinghai, this is the
most powerful strain found so far. This can be
transmitted from poultry to human and from human
to human. However, this strain of virus is unable
to transmit from human to poultry. The diagnosis
of this virus is extremely complicated.
Incubation period is short and causes of deaths
are mainly mis-diagnosis.The H5N1 from Qinghai
can kill laboratory chickens in 20 hours and lab
mice in 3 days.  As noted above and in additional
boxun reports, it can passes from human-to-human
with a short incubation period and an easily
mis-diagnosed clinical presentation.These
documents reveal a diverse gene pool of H5N1,
which is rapidly evolving via recombination. 
Human isolates from China have not been deposited
or even acknowledge by China.
HUMAN VIRUS
AVIAN VIRUS H5N1 Can spread between Bird
Cells Cannot Spread Between Humans
DEADLY REASSORTED VIRUS THAT CAN SPREAD BETWEEN
HUMANS. One Such Could Be H5N1 RK7
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CONTROL MECHANISMS
DETECTION/DIAGNOSIS The diagnostic yield of
different types of samples and virologic assays
is not well defined. In contrast to infections
with human influenza virus, throat samples may
have better yields than nasal samples. Rapid
antigen assays may help provide support for a
diagnosis of influenza A infection, but they have
poor negative predictive value and lack
specificity for influenza A (H5N1). The detection
of viral RNA in respiratory samples appears to
offer the greatest sensitivity for early
identification, but the sensitivity depends
heavily on the primers and assay method used.
Laboratory confirmation of influenza A (H5N1)
requires one or more of the following a positive
viral culture, a positive PCR assay for influenza
A (H5N1) RNA, a positive immunofluorescence test
for antigen with the use of monoclonal antibody
against H5, and at least a fourfold rise in
H5-specific antibody titer in paired serum
samples.
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CONTROL MECHANISMS
Antiviral Agents Patients with suspected
influenza A (H5N1) should promptly receive a
neuraminidase inhibitor pending the results of
diagnostic laboratory testing. The optimal dose
and duration of treatment with neuraminidase
inhibitors are uncertain, and currently approved
regimens likely represent the minimum required.
These viruses are susceptible in vitro to
oseltamivir and zanamivir. Oral osel-tamivir and
topical zanamivir are active in animal models of
influenza A (H5N1). Recent murine studies
indicate that as compared with an influenza A
(H5N1) strain from 1997, the strain isolated in
2004 requires higher oseltamivir doses and more
prolonged administration (eight days) to induce
similar antiviral effects and survival
rates.Inhaled zanamivir has not been studied in
cases of influenza A (H5N1) in humans.
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CONTROL MECHANISMS
Antiviral Agents (contd) Early treatment will
provide the greatest clinical benefit, although
the use of therapy is reasonable when there is a
likelihood of ongoing viral replication.
Placebo-controlled clinical studies of oral
oseltamivir and inhaled zanamivir comparing
currently approved doses with doses that are
twice as high found that the two doses had
similar tolerability but no consistent difference
in clinical or antiviral benefits in adults with
uncomplicated human influenza. Although approved
doses of oseltamivir (75 mg twice daily for five
days in adults and weight-adjusted twice-daily
doses for five days in children older than one
year of age twice-daily doses of 30 mg for
those weighing 15 kg or less, 45 mg for those
weighing more than 15 to 23 kg, 60 mg for those
weighing more than 23 to 40 kg, and 75 mg for
those weighing more than 40 kg) are reasonable
for treating early, mild cases of influenza A
(H5N1), higher doses (150 mg twice daily in
adults) and treatment for 7 to 10 days are
considerations in treating severe infections, but
prospective studies are needed. High-level
antiviral resistance to oseltamivir results from
the substitution of a single amino acid in N1
neuraminidase (His274Tyr). Such variants have
been detected in up to 16 percent of children
with human influenza A (H1N1) who have received
oseltamivir.Not surprisingly, this resistant
variant has been detected recently in several
patients with influenza A (H5N1) who were treated
with oseltamivir.Although less infectious in cell
culture and in animals than susceptible parental
virus, oseltamivir-resistant H1N1 variants are
transmissible in ferrets.Such variants retain
full susceptibility to zanamivir and partial
susceptibility to the investigational
neuraminidase inhibitor peramivir in vitro
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CONTROL MECHANISMS
Antiviral Agents (Contd) In contrast to
isolates from the 1997 outbreak, recent human
influenza A (H5N1) isolates are highly resistant
to the M2 inhibitors amantadine and rimantadine,
and consequently, these drugs do not have a
therapeutic role. Agents of clinical
investigational interest for treatment include
zanamivir, peramivir, long-acting topical
neuraminidase inhibitors, ribavirin,and possibly,
interferon alfa Immunomodulators
Corticosteroids have been used frequently in
treating patients with influenza A (H5N1), with
uncertain effects. Among five patients given
corticosteroids in 1997, two treated later in
their course for the fibroproliferative phase of
ARDS survived. In a randomized trial in Vietnam,
all four patients given dexamethasone died.
Interferon alfa possesses both antiviral and
immunomodulatory activities, but appropriately
controlled trials of immunomodulatory
interventions are needed before routine use is
recommended. Disposal of Birds More than
Hundreds of Millions of Birds Disposed.
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CONTROL MECHANISMS
Mechanisms of Oseltamivir Oseltamivir is an
antiviral drug, a neuraminidase inhibitor used in
the treatment and prophylaxis of both influenza A
and influenza B. Oseltamivir was the first orally
active neuraminidase inhibitor commercially
developed. Oseltamivir is a prodrug (usually
administered as phosphate), it is hydrolyzed
hepatically to the active metabolite, the free
carboxylate of oseltamivir (GS4071). Oseltamivir
acts as a transition-state analogue inhibitor of
influenza neuraminidase
(3R,4R,5S)-4-acetylamino-5-amino-3-
(1-ethylpropoxy)-1-cyclohexene-1-carboxylic acid
ethyl ester
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CONTROL MECHANISMS

• Contraindications of Oseltamivir
• The following information (but not its
  interpretation) comes from Roche's "Complete
  Product Information" publication for Tamiflu
  (intended for the United States).
• In the clinical trials performed by Roche
  (comparing roughly 2,700 individuals given
  Tamiflu with 2,650 given placebo), nausea and
  vomiting were the most frequent adverse reactions
  reported. Other adverse reactions were not
  reported by Tamiflu-treated patients at a
  markedly higher rate than those treated with
  placebo.
• According to Roche, in the postmarketing period,
  voluntary reports have possibly linked
  oseltamivir to the following other adverse
  reactions
• General Rash, swelling of face or tongue, toxic
  epidermal necrolysis
• Digestive Hepatitis, liver function tests
  abnormal
• Cardiac Arrhythmia
• Neurologic Seizure, confusion
• Metabolic Aggravation of diabetes
• Postmarketing studies are advantageous because
  the drug is effectively "tested" on a larger
  population, and previously missed adverse
  reactions may be discovered. However, given that
  forms are voluntary, it may be difficult to
  determine prevalency rates or whether an actual
  causal relation exists. The number of adverse
  reaction reports may be a clue, but these number
  are not reported by Roche in this document.

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CONTROL MECHANISMS
Contraindications of Oseltamivir
(Contd) Information from Japan neurological
effects and teen deaths In May 2004, the safety
division of Japan's health ministry ordered
changes to the literature accompanying
oseltamivir to add neurological and psychological
disorders as possible side effects, including
impaired consciousness, abnormal behavior, and
hallucinations. According to Japan's
Pharmaceuticals and Medical Devices Agency there
were 64 cases of psychological disorders linked
to the drug between fiscal years 2000 and 2004.
In February 2004, a 17-year-old male jumped in
front of a truck and died after taking one
capsule of Tamiflu. In February 2005, a
14-year-old male died after falling nine stories
from his condominium building. A third teen
reportedly attempted to jump from the window of a
building. The two deaths were reported to the
Japanese health ministry by Chugai Pharmaceutical
Co., a corporation half-owned by Roche which
distributes Tamiflu in Japan (Japan Times
November 13, 2005 Reuters Nov 14, 2005). Roche
points out that 32 million doses have been
prescribed worldwide, most of them in Japan, and
emphasizes the drug's safety. On November 18,
2005, a previously scheduled Advisory Committee
to the United States Food and Drug Administration
(FDA) met to reconsider the pediatric safety of
Tamiflu they issued a six-page report Pediatric
Safety Update for Tamiflu. The Committee stated
that there was insufficient evidence to claim a
causal link between oseltamivir use and the
deaths of 12 Japanese children (only two from
neurological problems). They did recommend adding
a warning to prescription information regarding
possible rashes. The author(s) of this section
have yet to find Japan's actual listing of
adverse reactions linked to oseltamivir. However,
it is known that one adverse reaction added to
the Japanese list was haemorrhagic Colitis
(bloody diarrhoea).
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CHALLENGES FACING CONTROL AND ERADICATION
TECHNIQUES 1. Viral Instability 2. The true
mechanism of successful vaccination a.      Lag
of phase in immune response as opposed to viral
interference and super-infection. 3. Stability of
Conquered Viruses.
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EVALUATING SAFETY OF NEW VACCINES HEPATITIS
AIDS SARS
21

• EVALUATING SAFETY OF NEW VACCINES
• Years of Development Safety Testing Needed As
  opposed to critical nature of current situation.
• Forced Coping Mechanism targeted against the
  modified host

22
Use of Known Agents In Interference Newcastle
Virus, Polio Virus Canine Distemper Feline
Panleucopenia Virus Caprine Arthritis
Encephalitis Virus (CAEV) Endogenous Cells
Protection Activation (ERV) Measles versus
Aids CAEV versus AVIAN flu Sense DNA / RNA
versus Anti-sense DNA/RNA Viral Infection
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The Use of Known Animal Vaccines To Create
Interference In Animals and Humans These can be
in raw and/or modified State Canine
Distemper Feline Panleucopenia Virus Caprine
Arthritis Encephalitis Virus (CAEV) Endogenous
Cells Protection Activation (ERV) Smallpox
Bacterial Viruses
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The Use of Known Animal Vaccines To Create
Interference In Animals and Humans Characteristi
cs such as positive DNA or RNA. CAEV versus
Avian Virus
ANTISENSE RNA
VIRUS
SENSE RNA
2
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The Use of Known Animal Vaccines To Create
Interference In Animals and Humans .
CAEV versus Avian Virus
26
The Use of Known Human Vaccines To Create
Interference In Animals and Humans Polio Smallpo
x Bacterial Viruses
27
The Use of Phages and Plasmids For Specific and
Non-Specific Interference and Immunity 200 Year
History
28
The Use of Chemical Agents To Interfere With
Viral Growth And Replication (Feed) BHT 2MEA AL7
21 Ethoxyquine, Detergent and pH modifiers

29
Resistant Condition Biological Membrane For
Concentration of Therapeutic Agent Cancer Rheumat
oid Arthritis Autoimmune Diseases Embryo
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Agent Includes Alpha-1 Antitrypsin, Sense /
Anti-sense Nucleic Acid Mono-clonal and
Poly-clonal Antibodies Transfer
Factor Interferons, Interleukins and Other
Modifying Anti-microbial Agents Universal
Antibiotic Regenerative Organ Protective
Mechanisms
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Universal Anti-microbial Agent This combines
all of the above mechanisms with super-imposed
structural analog including Methylthiols in
particular Bismuthiol that can occupy and protect
extra and intra-cellular sites attacked by the
microbial agent.
32
Colorized transmission electron micrograph of
H5N1 (golden) grown in Madin-Darby canine kidney
cells (green). (Source C. Goldsmith, J. Katz and
S. Zaki. Centers for Disease Control Prevention
Public Health Image Library. Image 1841.).
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Effective Immediate Therapeutic
Approach 1.           Vaccination With Active
Vaccines Based On Safe Tested Animal, Human
and Other Vaccines 2.           Supplementation
with Anti-Microbial Agent both in prevention
and therapy (feed augmentation) 3.           Long
Acting Passive Immunization With Universal
Vaccines
41
Advantages of Current Approaches Safety and
Efficacy Tested. CAEV Vaccines with Track
Record Supplements of Known Effects and
Side-Effects  
42
Results From Current Approaches 
Results From University of Stockholm
43
Results From Current Approaches 
Results As Tested And Reported By Cedars-Sinai
Medical Center In Los Angeles, California
44
Results From Current Approaches 
Continuation of Results (from prior slide) As
Tested And Reported By Cedars-Sinai Medical
Center In Los Angeles, California
45
Results From Current Approaches 
Results of efficacy as tested at Cedars-Sinai
Medical Center in Los Angeles California
46
Results From Current Approaches 
Results of Non-Toxicity as Shown and Tested at
Cedars-Sinai Medical Center in Los Angeles,
California
47
Results From Current Approaches 
Correspondence from Cedars-Sinai Medical Center
recognizing Profound Inhibition of Dr.
Chachouas Therapy
48
Results From Current Approaches 

• 30 year old lady
• presented with
• Tuberculosis
• Septicaemia.
• This resolved
• completely
• within two days
• of vaccination

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Results From Current Approaches 
Picture Showing Healthy Cell After Treatment of
Septicaemia
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Results From Current Approaches 

• Successful Treatment Confirmed By The
• University of Southern California

51
Results From Current Approaches 

• Successful Treatment Confirmed By The
• University of Southern California

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PRELIMINARY RESULTS CORONA VIRUS IN-VITRO CULTURES
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PRELIMINARY EVALUATION AVIAN FLU
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• Clinical Responses of Respiratory Distress
  Syndrome to Universal Vaccine
•  
• 30 cases of Status Asthmaticus
• 10 cases of Emphysema
• 20 cases of Pneumonia
• 10 cases of Cardio Vascular
• 12 cases of Neoplastic
• 5 cases of Tuberculous
• 3 cases Sarcoidosis
•  
• Subjective Improvement in virtually all cases
• Improvement in Spirometry significant in over 90
  of cases
• Efficacy decrease where Pleural Effusion involved
• UV in clinical use for over 20 years
• Safety tested and registered

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SUMMARY By the use of passive vaccines and
chemical agents as well as microbial
interference, 3 largely distinct therapeutic
modalities combine for effective prevention and
rapid, safe eradication of disease. No microbial
evolutionary mechanism can evolved rapidly if at
all to develop resistance against these three
distinct mechanisms. Vaccination with known
vaccines will both improve the current state of
health and in the previously immunized but
currently suffering from Avian flu, addition of
the human vaccines with the animal vaccine cannot
affect adversely a pre-vaccinated host. This is
an immediate and rapid solution for the
elimination of this and other epidemics.
56

• MECHANISM SUMMARY
• Genetic Interference
• Genetic, Extra-cellular and Intra-cellular
  Immunity and Repair
• Chemical and Microbial Interference.

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No living entity (microbe or cell) can easily
survive such a multi-directional approach. This
therapy originally developed for Cancer and Aids
but over 99 neutralization demonstrated against
multiple strains of Avian Flu in the lab setting
as well as Animal models and Respiratory Distress
Syndrome caused by any of the variety of
agents. As a by-product of research into
Cancer and Aids, we have a unique multi-prong
therapy against Avian Flu that offers real hope
in eliminating this epidemic and massively
reducing cases of Cancer Aids thus eliminating
not one but three of the greatest scourges of
mankind. As a by-product of research into
Cancer and Aids, we have a unique multi-prong
therapy against Avian Flu that offers real hope
in eliminating this epidemic and massively
reducing cases of Cancer Aids thus eliminating
not one but three of the greatest scourges of
mankind