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... characterization of HCV QS in vivo. Background HCV exists as quasispecies (QS) in infected individuals, consisting of a predominant viral variant and related, yet genetically distinct minor variants =-=[1]-=-. The study of HCV QS has historically focused on the hypervariable region 1 (HVR1) of the second envelope (E2) glycoprotein gene [2,3], the most variable region of the HCV genome. Early studies revea...

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...o the targets of: 82% of mothers to breastfeed their newborns in the early postpartum period, 61% and 34% continue breastfeeding at six months and at one year, respectively, as recommended by the AAP =-=[6]-=-. The fact that not all previous breastfeeding objectives were met despite the enormous effort put into breastfeeding promotion and support in the last decade suggests that pregnant and new mothers ma...

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...ization Survey 2004-2008, the CDC found that disparities exist among racial/ethnic groups, with the African-American population having one of the lowest rates of breastfeeding initiation and duration =-=[9]-=-. This is where another form of support may prove vital: community-based breastfeeding counselors (CBBCs). Community-based Counselors (CBCs) go by many names: peer educators, peer counselors, lay supp...

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... was developed as a means of reaching out to educate, support, advise, and counsel individuals in need of help who otherwise do not have access to the mainstream healthcare services using lay persons =-=[11]-=-. Whether that help relates to depression in seniors, diabetes management, surviving with HIV/AIDS, improving nutrition, recovering from alcohol abuse, coping with breast cancer, or any other health c...

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...itivity of E2-HVR1 PCR Assay To determine the sensitivity of the E2-HVR1 PCR assay using Taq versus HF-2 polymerases, serial dilutions of an HCV World Health Organization (WHO) international standard =-=[35]-=- were prepared from 50,000 to 50 International Units/milliliter (IU/ml). RNA was extracted, converted into cDNA and amplified by nested PCR as described above. PCR products were separated on agarose g...

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...epeatedly produced positive outcomes regarding breastfeeding, particularly with regards to increasing the rate of breastfeeding initiation and exclusivity, as well as extending breastfeeding duration =-=[19,21,23,24]-=-. In many cases, a CBBC is recruited from the target population/ community, which allows them to identify and reach people who may otherwise be isolated, underserved, or potentially unaware of the ser...

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...ys, including single strand conformation polymorphism analysis (SSCP) and heteroduplex mobility analysis (HMA) allow determination of HCV QS heterogeneity without the need for sequencing (reviewed in =-=[22]-=-). HMA was originally described for analyzing the sequence heterogeneity of the envelope gene of human immunodeficiency virus (HIV) [23,24]. HMA involves hybridization of a radioactive probe generated...

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...viral variant and related, yet genetically distinct minor variants [1]. The study of HCV QS has historically focused on the hypervariable region 1 (HVR1) of the second envelope (E2) glycoprotein gene =-=[2,3]-=-, the most variable region of the HCV genome. Early studies revealed that E2-HVR1 is a target of neutralizing antibodies [4-10]. Immune pressure is thought to be chiefly responsible for the fixation o...

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...in the HCV genome, has certain constraints in its structure. It is intriguing that key basic residues are highly conserved, which maintains the chemicophysical properties and conformation of the HVR1 =-=[33]-=-. Because HVR1 is an exposed domain on the E2 protein, these data suggest that the positively charged HVR1 is involved in interactions with negatively charged molecules such as lipids, proteins, or gl...

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...V QS heterogeneity without the need for sequencing (reviewed in [22]). HMA was originally described for analyzing the sequence heterogeneity of the envelope gene of human immunodeficiency virus (HIV) =-=[23,24]-=-. HMA involves hybridization of a radioactive probe generated from a QS variant to either heterogeneous PCR reaction products derived by direct PCR amplification from clinical specimens, or to homogen...

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...ation regarding benefits and techniques of breastfeeding passed along from CBBCs to new mothers, as well as making certain that the counseling techniques used by the CBBCs are appropriate and correct =-=[33]-=-. Through training, CBBCs need to acquire science-based practical knowledge about breastfeeding as well as be able to consider their own experiences and beliefs. These will contribute to the developme...

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...of the E2 gene. HCV QS have been analyzed in many different patient cohorts. HVR1 QS evolution may reflect progression of liver disease [11-15]. HCV QS also reflect the outcome of acute HCV infection =-=[16]-=-, and responses to antiviral therapy [17]. More recent studies have investigated the effect of HCV/HIV co-infection on HCV QS dynamics [18,19]. In the current study, we analyzed 20 baseline samples fr...

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...d in most studies of HCV QS analysis. However, given that Taq lacks proofreading activity, mathematical debates have been raised to suggest that QS evolution is overestimated by Taq induced mutations =-=[30]-=-. Indeed, in one report, use of Taq increased the proportion of minor QS variants [31], and we have found that, in general, QS diversity is lower if proof-reading enzymes are used instead of Taq [32]....

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... understanding of the needs of the target population,” which should include the actual physiology of breastfeeding, hands-on support techniques, positive counseling skills, and cultural understanding =-=[27]-=-. As mentioned previously, breastfeeding is an area in which CBBCs have proven to be very effective; however, there is a lack of consistency in training curriculum from program to program. There are d...

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...search purposes. Bronner et al conducted a national survey of WIC, one of the largest employers of breastfeeding counselors, regarding their role at the state and local levels within the organization =-=[32]-=-. Although the findings of Bronner et al support other studies that have established the effectiveness of CBBCs in breastfeeding promotion, they found that there was a lack of consistency in the recru...

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...-15]. HCV QS also reflect the outcome of acute HCV infection [16], and responses to antiviral therapy [17]. More recent studies have investigated the effect of HCV/HIV co-infection on HCV QS dynamics =-=[18,19]-=-. In the current study, we analyzed 20 baseline samples from the HALT-C trial and 12 HCV-HIV co-infected patient samples. The HALT-C study is a randomized multi-center clinical trial to assess the eff...

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...m the target population/ community, which allows them to identify and reach people who may otherwise be isolated, underserved, or potentially unaware of the services and information available to them =-=[15]-=-. Sufficient training is imperative to a successful program with positive health outcomes, and in the case of breastfeeding, increasing the proportion of mothers initiating, as well as extending, the ...

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...med a systematic review of randomized controlled trials from around the world that evaluated the impact of CBBC on breastfeeding initiation, rates, exclusivity, and maternal and child health outcomes =-=[22]-=-. In their review, the researchers found that interventions that included antenatal and multiple perinatal CBBC contact yielded the most success in increasing breastfeeding initiation rates as compare...

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...t breastfeeding as well as be able to consider their own experiences and beliefs. These will contribute to the development of their interpersonal skills that they will use when counseling new mothers =-=[34]-=-. While extensive research has been done to support the importance of CBBCs, to date there has not been a comprehensive nationwide study to assess the impact of CBBC attributes, such as CBBC-education...

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...viral variant and related, yet genetically distinct minor variants [1]. The study of HCV QS has historically focused on the hypervariable region 1 (HVR1) of the second envelope (E2) glycoprotein gene =-=[2,3]-=-, the most variable region of the HCV genome. Early studies revealed that E2-HVR1 is a target of neutralizing antibodies [4-10]. Immune pressure is thought to be chiefly responsible for the fixation o...

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...ity, mathematical debates have been raised to suggest that QS evolution is overestimated by Taq induced mutations [30]. Indeed, in one report, use of Taq increased the proportion of minor QS variants =-=[31]-=-, and we have found that, in general, QS diversity is lower if proof-reading enzymes are used instead of Taq [32]. But the question arises as to whether Taq induced mutations affect the outcomes/concl...

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...eastfeed. In 2005, Anderson et al published results from a randomized, controlled trial showing that CBBC positively influences breastfeeding outcomes among low-income Latina and other minority women =-=[21]-=-. Chapman et al performed a systematic review of randomized controlled trials from around the world that evaluated the impact of CBBC on breastfeeding initiation, rates, exclusivity, and maternal and ...

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...d if inadequate training is provided [17]. When counseling does not provide capable support in breastfeeding, there is a higher risk of failure in implementing good breastfeeding practices by mothers =-=[28]-=-. The World Health Organization (WHO) set the goldstandard for CBBC training programs at 40 hours duration, although they recommend a minimum of 18 hours with an additional 3 hours of hands-on practic...

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...on-α (peg-IFN) therapy on the progression of liver fibrosis and development of decompensated liver disease in hepatitis C patients who are non-responders to prior pegylated IFN plus ribavirin therapy =-=[20,21]-=-. Viral QS have been analyzed by many techniques. Cloning and sequencing is the gold standard. Electrophoretic mobility-based assays, including single strand conformation polymorphism analysis (SSCP) ...

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...ment [33]. In this regard, it has recently been shown that high pretreatment HCV QS diversity and complexity at baseline are associated with non-response to pegylated IFN ribavirin in HALT-C patients =-=[34]-=-. HAART therapy appeared to increase HCV QS diversity and complexity, regardless of the enzyme used, but this did not reach statistical signficance in this small cohort. These data are consistent with...

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...ed Food and Nutrition Education Program (EFNEP); the Special Supplemental Nutrition Program for Women, Infants, and Children (WIC); and the Breastfeeding Heritage and Pride Program with great success =-=[19,20]-=-. These communitybased breastfeeding counselors have been used extensively and effectively to promote breastfeeding in selected communities, especially low-income minority communities who otherwise do...

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...ison of the heterogenous PCR product ("H") versus CFA gel shift patterns. If the gel shift patterns of the individual clones are representative of the heterogenous PCR product, the CFA was successful =-=[11,26,27]-=-. As shown in Figure 2A, patient 9 had very few discernible gel shifts, and the shifts themselves were not very distinct from the homoduplex probe control. The same pattern was observed when either Ta...

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...-15]. HCV QS also reflect the outcome of acute HCV infection [16], and responses to antiviral therapy [17]. More recent studies have investigated the effect of HCV/HIV co-infection on HCV QS dynamics =-=[18,19]-=-. In the current study, we analyzed 20 baseline samples from the HALT-C trial and 12 HCV-HIV co-infected patient samples. The HALT-C study is a randomized multi-center clinical trial to assess the eff...

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...on-α (peg-IFN) therapy on the progression of liver fibrosis and development of decompensated liver disease in hepatitis C patients who are non-responders to prior pegylated IFN plus ribavirin therapy =-=[20,21]-=-. Viral QS have been analyzed by many techniques. Cloning and sequencing is the gold standard. Electrophoretic mobility-based assays, including single strand conformation polymorphism analysis (SSCP) ...

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...V QS heterogeneity without the need for sequencing (reviewed in [22]). HMA was originally described for analyzing the sequence heterogeneity of the envelope gene of human immunodeficiency virus (HIV) =-=[23,24]-=-. HMA involves hybridization of a radioactive probe generated from a QS variant to either heterogeneous PCR reaction products derived by direct PCR amplification from clinical specimens, or to homogen...

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...ther heterogeneous PCR reaction products derived by direct PCR amplification from clinical specimens, or to homogeneous HVR1 sequences derived from cloned QS variants (clonal frequency analysis, CFA; =-=[25]-=-). Hybridizations between the probe and various target sequences result in the formation of double stranded DNA molecules (heteroduplexes) that produce shifts when the hybrids are separated on non-den...

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...ison of the heterogenous PCR product ("H") versus CFA gel shift patterns. If the gel shift patterns of the individual clones are representative of the heterogenous PCR product, the CFA was successful =-=[11,26,27]-=-. As shown in Figure 2A, patient 9 had very few discernible gel shifts, and the shifts themselves were not very distinct from the homoduplex probe control. The same pattern was observed when either Ta...

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...ison of the heterogenous PCR product ("H") versus CFA gel shift patterns. If the gel shift patterns of the individual clones are representative of the heterogenous PCR product, the CFA was successful =-=[11,26,27]-=-. As shown in Figure 2A, patient 9 had very few discernible gel shifts, and the shifts themselves were not very distinct from the homoduplex probe control. The same pattern was observed when either Ta...

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... [30]. Indeed, in one report, use of Taq increased the proportion of minor QS variants [31], and we have found that, in general, QS diversity is lower if proof-reading enzymes are used instead of Taq =-=[32]-=-. But the question arises as to whether Taq induced mutations affect the outcomes/conclusions in cohort-based studies on HCV QS. Thus, in the current study, we compared Taq and HF-2 polymerases in ter...

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...astfeeding supporters, the United States can only boast of approximately 11,000 who are limited to certified Baby Friendly Hospitals and birthing facilities and just a few non-Baby Friendly Hospitals =-=[7,8]-=-. This means they may not be able to reach all populations, particularly those most in need (low-income, immigrants/minorities and residing in deprived communities). In the National Immunization Surve...

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...ing counselors (CBBCs). Community-based Counselors (CBCs) go by many names: peer educators, peer counselors, lay support, community outreach workers, and indigenous health care advisors, among others =-=[10]-=-. The concept of CBC was developed as a means of reaching out to educate, support, advise, and counsel individuals in need of help who otherwise do not have access to the mainstream healthcare service...

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... curriculum from program to program. There are differences in knowledge and proficiency between a skilled professional and a peer educator, which can be exacerbated if inadequate training is provided =-=[17]-=-. When counseling does not provide capable support in breastfeeding, there is a higher risk of failure in implementing good breastfeeding practices by mothers [28]. The World Health Organization (WHO)...

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...ed Food and Nutrition Education Program (EFNEP); the Special Supplemental Nutrition Program for Women, Infants, and Children (WIC); and the Breastfeeding Heritage and Pride Program with great success =-=[19,20]-=-. These communitybased breastfeeding counselors have been used extensively and effectively to promote breastfeeding in selected communities, especially low-income minority communities who otherwise do...

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...The World Health Organization (WHO) set the goldstandard for CBBC training programs at 40 hours duration, although they recommend a minimum of 18 hours with an additional 3 hours of hands-on practice =-=[29]-=-. This suggests the importance of hands-on practice in the training of CBBCs. Although both the California Department of Health’s WIC-based breastfeeding peer counseling training program and the one o...

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...seling training program and the one offered by La Leche League International are approximately the same in duration at around 20 hours, content can vary with location, program, state, or even country =-=[30,31]-=-. Rossman and Chapman et al in their systematic reviews of numerous CBBC studies/programs, demonstrated the variability among the training protocols used by different programs [22,27]. It was determin...