Hepatitis C Virus Ebook

Alternative Hepatitis C Treatments

The therapeutic goals of Natural treatment for Hepatitis C are as follows: Decrease iral load Normalize liver enzyme levels. Enhance/regulate immune system function. Strengthen and promote healthy liver function. Protect the liver, prevent further damage. Virological response; i.e. viral clearance, viral reduction or elimination of the virus. Starve the virus by limiting levels of iron. Optimizing cellular levels of glutathione in the body, making detoxification of the liver possible and enhancing the immune system. Stimulate regeneration of the damaged liver cells. Use of antioxidants to combat the effects of free-radicals generated by the virus. Reduce inflammation. Slow viral replication. Replace all of the inflammation-damaged liver cells. Regulate immune function/prevent auto-immune problems. Cancer preventative measures. Reverse fibrosis to prevent and improve cirrhosis

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Inhibition of HCV by RNAi

HCV is a major cause of chronic hepatitis and hepatocellular carcinoma. Currently, no vaccines are available for HCV, and several groups have used RNAi to target HCV replication. HCV belongs to the family Flaviviridae, and its genome is encoded by a 9.6-kb RNA of positive polarity. Because there is no cell culture system for HCV replication, all studies have used replicon systems in Huh-7 cells as a model for HCV replication. These replicons support HCV RNA transcription and protein synthesis, but do not produce infectious virus. Regions that have been targeted include conserved sequences in the capsid, NS3, NS4A B, NS5B, and the 5'-UTR (see Table 3). The 5'-UTR of the HCV RNA is a good potential target because it is the most conserved part of the HCV genome that harbours the internal ribosomal entry site (IRES), which is required for translation. Researchers have used siRNAs and shRNAs to block HCV replication (Takigawa et al. 2004). In addition, they looked at virus replication in...

Viral Hepatitis

Another major hepatic pathogen is HCV (see above), which also cannot easily be grown in cell culture. For this reason, plasmid replicons containing subgenomic fragments or the whole genome of HCV are used. To show the general applicability of antiviral RNAi approaches in mice, McCaffrey and colleagues (2002) used high-pressure tail vein injection to deliver a plasmid expressing a fusion construct of luciferase and the viral protein NS5B to the livers of mice. When siRNAs or shRNA-expressing plasmids were co-transfected with the reporter plasmid, 80 and 90 reduction of luciferase expression over the whole body of the animals were measured, respectively. Several groups have reported on siRNAs directed against different regions on the mRNA of HCV that reduced viral RNA and protein levels (for a listing, see Radhakrishnan et al. 2004 Randall and Rice 2004). Randall and colleagues (2003) found a greater than 98 inhibition of RNA levels on transfection of an efficient siRNA. This decrease...

Using RNAi to Treat Other Viruses

Although many previous studies on RNAi-mediated inhibition have focused on HIV-1, there is a growing body of data addressing the inhibition of other animal and human viruses. These include RNA viruses such as hepatitis C virus (HCV), poliovirus, Semliki Forest virus (SFV), influenza virus A, rhesus rotavirus (RRV), and Rous sarcoma virus (RSV), and DNA viruses such as human papillomavirus type 16 (HPV-16) and hepatitis B virus (HBV). In most of these studies, the RNAi machinery was directly targeted towards the viral RNA using synthetic siRNAs. Hepatitis induced by HBV or HCV is a major health problem. At present, hundreds of millions of individuals are infected worldwide. Although there is an effective vaccine against HBV, it is only useful for the prevention of viral infection. There is no vaccine for HCV. Hepatitis caused by these two viruses has therefore been an important target for potential RNAi therapy. HCV is a major cause of chronic liver disease, which can lead to liver...

Coagulation Factor Transfusion

On finding of blood donor and donor selection should be mentioned. In addition, the problem of blood borne infection in the present day should be stated. Human immunodeficiency virus (HIV), hepatitis B virus (HBV) and hepatitis C virus (HCV) in the blood supply are the important problematic blood borne transmitted diseases. In general, cryoprecipitate transfusion is the widely used coagulation factor transfusion for the cases with hemophilia 46-48 . According to the study of Evatt et al., over a lifetime of treatment (60 years), the cumulative risk of HIV exposure for a person with hemophilia getting monthly infusion of cryoprecipitate prepared from plasma is significant, 2 in the USA and 40 in Venezuela 49 . Considering the cumulative risk for transmitting HIV to patients with hemophilia through cryoprecipitate treatment, medical care providers should carefully evaluate the use of cryoprecipitate in any but emergency conditions or when no virally inactivated outcomes are available 47...

Risk of Transfusions

Antigens during a previous transfusion which results in a reactive action 3 to 10 days after a subsequent transfusion) are the two most common complicated outcomes of blood transfusion 4 . The third kind is febrile, non-hemolytic owing to reactive action between antibodies as well as LEUKOCYTE antigens or plasma proteins 5 . It should be noted that all three kinds are rare in the newborn because of a defect in isohemagglutins as well as poor antibody response to alien antigens. In addition, blood transfusion also had the risk for transmission of many blood borne pathogenic agents 6 including syphilis, HIV 1, HIV 2, Human T-lymphotrophic virus 1 (HTLV-1), hepatitis B surface antigen (HbsAg),malaria, microfilaria as well as babesia. These pathogenic agents should be screened in the basic blood transfusion screening test. Also, some new biomarkers are introduced including hepatitis C virus (HCV), alanine aminotransferase (ALT), a surrogate biomarker for HIV infection, as well as...

Inhibition of Viruses by RNAi

The first demonstration of RNAi-mediated inhibition of a human pathogenic virus was reported by Bitko and Barik in 2001 (Bitko and Barik 2001). These authors reported a tenfold inhibition of human respiratory syncytial virus (HRSV) replication in vitro using nanomolar concentrations of synthetic siR-NAs that targeted the viral polymerase subunit P and the fusion protein F. Currently, many other studies have described RNAi-mediated inhibition of a large variety of viruses. RNAi-mediated inhibition of HIV-1 has received much attention (see below, Tables 1 and 2). In addition, 17 different RNA viruses, and 10 different DNA viruses have been efficiently targeted by RNAi (Tables 3 and 4). These include important human pathogens such as hepatitis C virus (HCV), dengue (DEN) virus, severe acute respiratory syndrome (SARS) coronavirus, poliovirus, influenza A virus, hepatitis D virus (HDV), human rhinovirus-16 (HRV-16), hepatitis B virus (HBV), herpes simplex virus type-1 (HSV-1), human...

Diversity of Viral Targets

Viruses other than HIV-1 have also been successfully targeted by siRNAs in vitro with some success, including Semliki forest virus (SFV), poliovirus, dengue virus, influenza virus, hepatitis C virus, and many others (reviewed in Radhakrishnan et al. 2004). The fact that such a wide berth of varying viruses can be successfully targeted by siRNAs suggests that these nucleic acid molecules can be used to theoretically target virtually any emerging or present-day infectious agent. However, despite the excitement and the early proofs-of-principle in the literature, there are important issues and concerns about therapeutic application of this technology, including difficulties with efficient delivery, uncertainty about potential toxicity, and the emergence of siRNA-resistant viruses. In particular, certain viruses encode proteins that block one or more steps in the RNAi pathway (Bennasser et al. 2005 Hamilton et al. 2002 Johansen and Carrington 2001 Li et al. 2002 Llave et al. 2000 Mallory...

Cleavage of Viral RNA by Ribozymes

In addition to HIV-1, hepatitis B and C viruses have been major targets of antiviral ribozymes. For example, a chemically stabilized hammerhead ri-bozyme has been designed that targets the highly conserved 5'-UTR of HCV. In cell culture, it inhibited replication of an HCV poliovirus chimera by up to 90 (Macejak et al. 2000). This ribozyme, dubbed Heptazyme, has subsequently been employed in clinical trials as will be outlined below. A different approach has been developed to generate a ribozyme targeting HBV by expression of a triple-ribozyme cassette that undergoes intracellular self-processing (Pan et al. 2004). This sophisticated construct was suitable to reduce viral DNA in the liver of transgenic mice by greater than 80 , as measured 2 weeks after infection. Furthermore, a DNAzyme containing phosphoroamidate nucleotides was capable of inhibiting the replication of influenza A viruses by more than 99 (Takahashi et al. 2004).

Introduction to Vaccine [

Vaccination is classified as an active immunization. This means tit makes uses of human beings physiological process to create immune by itself. There are several vaccine at present. Recommendation for vaccination might be different in different settings. Expanded Program on Immunization (EPI) is the basic public health one in all countries. With use of vaccine, controls of many infectious diseases are feasible. Pox is an example of infectious disease that can be controlled by vaccine. This infection is already got rid of by succeed in global vaccination. The presently widely used vaccine include rabies vaccine, measles vaccine, rubella vaccine, Japanese encephalitis vaccine, hepatitis B vaccine, hepatitis A vaccine, influenza vaccine, varicella vaccine, pneumococcal vaccine, pertussis vaccine, diphtheria vaccine, BCG and toxiod. However, at present, there are attempts to develop more vaccines for real clinical usage . This is the aim of preventive medicine. In addition, there are...

Ribozymes and DNAzymes

Ribozymes and DNAzymes are oligonucleotides with catalytic activity. Ri-bozymes are made of ribonucleotides, whereas DNAzymes are composed of deoxyribonucleotides, and both can be designed to target specific transcripts. Upon hybridizing to the target mRNA, they cleave the transcript in a sequence-specific fashion (Fig. 1). Ribozymes have been more extensively investigated than DNAzymes. The results of these studies show that ribozymes can cleave phosphodiester bonds without the aid of protein enzymes (Jen and Gewirtz 2000), and that the catalytic moiety of ribozymes recognizes a specific nu-cleotide sequence, GUX, where X C, U, or A (Ruffner et al. 1989) or in some cases, NUX, where N is any nucleotide (Xing and Whitton 1992). Hammerhead, hairpin, group I intron, ribonuclease P, and the hepatitis delta virus ribozymes are five naturally occurring ribozymes that are used as the basis for the catalytic motifs of engineered ribozymes. A greater emphasis has been placed on studying...

Other RNA Molecules Interacting with Aminoglycosides

Coworkers (von Ahsen et al. 1991) demonstrated that aminoglycosides bind and inhibit group I intron self-splicing (for a review see Schroeder et al. 2000). Given the existence of self-splicing RNA group I introns in bacteria, these are indeed potential drug targets to explore. This is also apparent based on the knowledge that group I introns have not been identified in mammalian cells. Other ribozymes known to be inhibited by aminoglycosides are the self-cleaving hammerhead ribozyme, the hairpin ribozyme (Stage et al. 1995 Earnshaw and Gait 1998), and the self-cleaving hepatitis delta virus HDV RNA (Rogers et al. 1996 Chia et al. 1997). In this context, we note that the aminoglycoside streptomycin is apparently a better inhibitor of nuclear pre-mRNA splicing compared to inhibition of the group I intron splicing (Hertweck et al. 2002). Since no other aminoglycosides have been tested for inhibition of nuclear pre-mRNA splicing, it would be of value to study this in more detail. This is...

Antisense Oligonucleotides as Antiviral Agents

Infections with HCV are a major health problem worldwide. Chronic infection with this plus-stranded RNA virus causes liver cirrhosis, liver failure, and hepatocellular carcinoma, often leading to the requirement of liver transplantation. Since current treatment of HCV is unsatisfactory, a need for new, specific anti-HCV drugs stands to reason. AS ONs have therefore widely been used with the intention of inhibiting HCV replication. The 5'-untranslated region (UTR) is one of the most highly conserved regions of the HCV genome and has most frequently been targeted with AS ONs. Since this region is strongly structured, intensive efforts were made to identify accessible target sites. The entire viral cycle of HCV is cytoplasmic and thus AS ONs do not necessarily need to be designed to activate RNase H, which is mainly located in the nucleus. Rather, it has been shown that AS ONs interfering with the assembly of a translation initiation complex on the internal ribosome entry site (IRES)...

From Bench to Bedside

A number of companies like Alnylam Pharmaceuticals, Benitec, International Therapeutics, CytRx Corporation, Sirna Therapeutics, and Nucleonics are developing siRNAs for the treatment of viral infections such as HIV, HCV, and HBV, according to the companies' respective Web sites. Two companies, Sirna Therapeutics and Acuity Pharmaceuticals, have recently initiated phase I clinical studies of siRNAs interfering with vascularization for treatment of age-related macular degeneration. Results from these studies would be the first clinical data available for siRNAs and may allow better-founded predictions on the future of RNAi in therapeutic approaches.

Inhibition of HBV by RNAi

Targeting the small HBV surface antigen, HbsAg (Giladi et al. 2003). Earlier, a 6-fold inhibition of HBV in the liver of mice was obtained by intravenous injection of plasmids expressing shRNAs against the core, HbsAg Pol and X gene (McCaffrey et al. 2003). Possibly, a similar approach could be used in infected patients to lower virus titres. Besides targeting the viral RNA, it is also possible to inhibit HBV replication by targeting the mRNAs of cellular factors that are required for virus replication. Ni et al. have shown that targeting the hLa protein results in a decrease in virus replication (Ni et al. 2004). Additionally, it has been shown that RNAi can be used to prevent HCV- or HBV-induced disease of the liver by silencing the expression of the cellular Fas gene. During HCV or HBV infection Fas-mediated apoptosis of hepa-tocytes is triggered as a self-destructive inflammatory response of the liver. Silencing Fas expression with synthetic siRNAs blocks this reponse (Li et al....

Ribozymes in Clinical Trials

In addition to these studies to treat patients with HIV infections, one clinical trial was initiated with a hammerhead ribozyme targeting HCV. In contrast to the trials described in the preceding paragraph, in which viral vectors were employed to deliver ribozymes, the aforementioned chemically presynthesized ribozyme Heptazyme was used for these tests (Usman and Blatt 2000). Encouraging results of initial clinical studies led to the initiation of a phase II trial with Heptazyme alone and in combination with interferon (IFN) in 2001. The finding that the HCV RNA level in the serum of patients treated with the ribozyme was reduced by only 10 , along with results from toxological studies, led to the decision to stop clinical experimentation of Heptazyme (Peracchi

HLAB and susceptibility to ankylosing spondylitis

By the early 1970s there were growing numbers of reports of association between the possession of particular HLA antigens on the surface of white blood cells and susceptibility to disease, notably coeliac disease, multiple myeloma, lymphoma, systemic lupus erythematosus, and psoriasis. During the course of the year in 1972, independently reproducible and highly significant associations were found between the possession of HL-A1 and HL-A8 and coeliac disease (Falchuk et al. 1972 Stokes et al. 1972), W17 and HL-A13 and psoriasis (Russell et al. 1972), and HL-A1 and HL-A8 and chronic active hepatitis (Mackay and Morris 1972). The following year there were dramatic reports of an HLA association with a chronic

The Cytometric Bead Array System

Bead Position Cba

Multiplex assays are well suited to demonstrate a pattern of antibody responses against infectious agents, thus providing a bead assay analog to the Western blot method. McHugh (10) presented a prototype Hepatitis C virus antibody assay for potential use in the blood bank. Faucher (11) has demonstrated the capability of the multiplex assays to detect HIV-1 antibodies from fresh plasma and from dried bloodspot specimens. Khan (12) utilized the multiplex format to construct a serological assay, which detected 10 highly prevalent mouse infectious pathogens in a single reaction.

Genetic diversity in Hla Kir and HIV strategies for survival

And is the preferred site of reciprocal recombination of the centromeric and telomeric motifs leading to further haplotypic diversity (Martin et al. 2003). HLA-A molecules such as A3 and A11 are recognized by receptors encoded by KIR3DL2 and KIR2DS1 at the telomeric end of the KIR locus, HLA-B Bw4 is a ligand for KIR3DL1, and in the centromeric part receptors for HLA-C molecules are encoded. HLA-C1 allotypes with Asn at position 80 are recognized by receptors encoded by KIR2DL3 and KIR2DL2 KIR2DL1 encodes KIR receptors with a methionine rather than lysine at position 44 and these recognize HLA-C2 allotypes with lysine at position 80. As well as the disease associations with HIV-1 infection, genetic diversity at the KIR and HLA class I is important in other infections, autoimmunity, reproduction, and cancer. For example, maternal KIR genotype and fetal HLA-C type are important in pre-eclampsia (Hiby et al. 2004) homozygosity of KIR2DL3 and of HLA-C1 allotypes modulate hepatitis C...

Polymorphism haplotypes and disease

The much higher than expected rate of nonsynony-mous (amino acid changing) nucleotide substitutions within coding regions for peptide binding in MHC molecules compared to other areas of the MHC is consistent with Darwinian selection operating at these gene loci (Hughes and Nei 1988, 1989). Analysis of HLA-A and -B showed significantly less homozygosity than expected given neutrality (Hedrick and Thomson 1983), with amino acid heterozygosity concentrated in peptide binding regions when diversity across different human populations was studied (Hedrick et al. 1991). Finding clear evidence of heterozygote advantage in terms of disease susceptibility has, however, been surprisingly difficult. Examples have been found for a number of viral infections. Maximal heterozygosity at HLA-A, -B, and -C loci has been associated with delayed onset of acquired immuodeficiency syndrome (AIDS) and lower mortality in HIV-1 infection (Section 14.4) (Carrington et al. 1999). Heterozygosity of class II...

Using Antibodies In Research And Medicine

Camel Antibody

Camel antibodies are only the most recent in a long line of medical and biological applications of antibodies. One of the main uses is to detect diseases in patients, a process known as serology. Doctors take a sample of blood and measure whether it contains antibodies against particular infectious agents. High amounts of IgM antibodies are a sign that a person is infected with a particular virus or has been infected very recently. Antibodies are also used to look for signs of hepatitis or other liver diseases, including some autoimmune conditions. In pregnancy

Cytokine Treatment

Cytokine treatment makes use of administration of cytokine for curative purpose. This is famous for a long time for many diseases. Good examples are hepatitis virus infections. Important interesting reports on cytokine treatment will be given as idea to the readers in Table 3. Badr et al. suggested that poly-functionality of HCV-specific T cells could be predictive of the outcome of acute HCV and that early therapeutic intervention, IFN-alpha, antiviral therapy, could reconstitute the pool of long-lived poly-functional memory T cells 77 . Bolewska et al. studied on IFN alpha, gamma, omega before and during treatment of chronic hepatitis C with pegylated interferon alpha and ribavirin 78 . Bolewska et al. found that there was no correlation in IFN alpha, gamma and omega concentrations with efficacy of antiviral treatment 78 . Adamek et al. studied on long-term viral response to IFN alpha 2b plus ribavirin in chronic HCV patients during standard therapy

RNA Viruses

The genome of RNA viruses has also proved a sensitive target for PNA. Whereas the ribosome is a very robust cellular machinery that apparently has evolved to unravel folded mRNA structures and therefore is usually not severely affected by bound antisense agents, reverse transcriptase is very sensitive to blockage by e.g. PNA bound to the virus RNA template (Chaubey et al. 2005). There may be good opportunity to exploit this finding in the discovery of novel drugs in the combat of acquired immunodeficiency syndrome (AIDS). Hepatitis B and C viruses have also been successfully targeted at the RNA level using PNA antisense in vitro and in cell culture systems (Robaczewska et al. 2005 Nulf and Corey 2004).

Catalytic Regulation

The finding by Cech and colleagues that an intron from Tetrahymena ther-mophila (Cech et al. 1981) could catalyze the cleavage and joining of olig-oribonucleotide substrates in a highly specific manner confirmed previous suspicions that RNA could act as a robust catalyst. Since then, ribozymes with a variety of reaction mechanisms and functions have been discovered or evolved (Doudna and Cech 2002). In particular, the hammerhead ribozyme, at roughly 40 nt, is one of the smallest naturally occurring ribozymes and participates in the rolling circle replication of some viroids by carrying out a self-cleavage reaction in which a 2' oxygen participates in the attack on the scissile phosphate. The crystal structure of the hammerhead (Pley et al. 1994) revealed an active site formed by the apposition of three short helices, and catalysis occurs at the conserved trihelical junction. Whether hammerhead catalysis requires divalent cations and is driven by global or local rearrangements of the...

Viral Escape

We have recently described an siRNA double expression vector (SiDEx) containing genes for two different siRNAs both targeted against the RNA-dependent RNA polymerase of coxsackievirus B3 (Schubert et al. 2005a Fig. 4). Even when an artificial point mutation was introduced into the target gene so that the respective mono-expression vector was no longer capable of silencing, the double expression vector maintained high silencing activity. Although we did not find any deleterious effect of the simultaneous use of two siRNAs, it has been observed that different siRNAs present in one cell may compete for access to the RNAi machinery (Bitko et al. 2005). Thus, the efficiency of one siRNA may be compromised due to the presence of a second species. Another difficulty in preventing viral escape may be the amount of viral particles present in the blood of patients in the state of viremia. In HCV infection, up to 107 virions per milliliter may be present in the bloodstream, so...

Line strength

Both line positions and line strengths must be understood in order to interpret the structure of observed spectra and derive abundances and temperatures of contributing molecules. Following Planck (1901) and Einstein (1906b), we treat radiation as composed of photons with energy E hcv, where h is the Planck constant and cv is the frequency of the radiation in hertz. The intensity is proportional to the arrival rate of photons. If these photons originate from molecules in a small volume, the intensity is proportional to the number of molecules in the optical path undergoing transitions at that frequency.

Neonatal Pathology

Figure 16.6. (a) Gross necropsy (liver) of neonate with septicaemia and hepatitis caused by Staphylococcus intermedius. (b) Lungs (L) of a neonate that acquired pneumonia prenatally from a dam with an ascending urogenital tract infection that caused chorioamnionitis. (c) Microscopically the pulmonary alveolar spaces contained inflammatory cells (arrow). Pseudomonas aeruginosa was cultured from the lungs and P. fluorescens from heart blood. (See also Plate XIX.) Figure 16.6. (a) Gross necropsy (liver) of neonate with septicaemia and hepatitis caused by Staphylococcus intermedius. (b) Lungs (L) of a neonate that acquired pneumonia prenatally from a dam with an ascending urogenital tract infection that caused chorioamnionitis. (c) Microscopically the pulmonary alveolar spaces contained inflammatory cells (arrow). Pseudomonas aeruginosa was cultured from the lungs and P. fluorescens from heart blood. (See also Plate XIX.)

Biological Hazards

Equally important for the team members, beyond researching the CDC and WHO information, is to consult their primary care physicians and a physician who specializes in tropical medicine. It is critical to make arrangements to see these physicians well in advance of the trip, as appointment schedules often need to be made months in advance. In addition, if immunizations are required, appropriate lead time is required for many immunizations to be effective. Common immunizations may include those for yellow fever, tetanus update, hepatitis A, typhoid, and others dictated by the destination. Commonly required medications may include those for malaria prevention and prophylactic treatment for altitude sickness. In addition, tropical medicine physicians will often prescribe appropriate antibiotics to be taken should a team member contract gastrointestinal infections.