Group G-D was inoculated with 107 C6/36 derived RVFV. Group G-E was inoculated with 105 PFU of Vero E6 RVFV stock, and re-inoculated IV with the same inoculum at 1 dpi. Group G-F was inoculated with 105 PFU of C6/36 derived RVFV, and re-inoculated

IV with the same inoculum at 1 dpi. Group G-G was inoculated with 107 C6/36 derived RVFV and re-inoculated SC with the same inoculum at 1 dpi. All goats were kept for four weeks following the inoculation to monitor an antibody development. Serum samples collected at 0, 4, 5, 6, 7, 14, 21 and 28–30 dpi were analyzed for presence of neutralizing antibodies. Differences in susceptibility to RVFV infections were observed between sheep and goats, and also between breeds of sheep. In the first study,

conducted JAK2 inhibitors clinical trials in Suffolk-cross sheep, all animals developed viremia at 3 dpi, both by virus isolation and RNA detection when inoculated with 105 PFU of virus produced in Vero cells. However, when the Rideau Arcott cross lambs were inoculated via the same route and the same inoculum, only three out of four animals had detectable RVFV RNA in their blood and only two developed viremia (Fig. 1). Subsequently different inoculation approaches were tested to obtain a more reliable viremia model. Genomic sequences of the inocula were verified prior to the start of the animal inoculations. Concurrently with the infection experiments, characterization on protein level of RVFV NSC 683864 manufacturer generated in Vero E6 cells or the C6/36 was taking place. There was no difference in genome of RVFV generated in Vero E6 cells

compared to virus generated in C6/36 cells, including the stock viruses used in experimental medroxyprogesterone inoculations, and the sequences corresponded with sequences published for RVFV ZH501 in Gen Bank. Both viruses had functional NSm and NSs coding genes, as immunoblots of infected cell lysates indicated that all proteins from the M and S segments were expressed. The viruses however differed in protein composition of virions, with the mosquito-cell generated RVFV having an additional large glycoprotein (78 kDa) incorporated into virions [23]. Subcutanous inoculation was used in all primary inoculation. Two doses (105 or 107 PFU/animal) and two different inocula (prepared either in Vero E6 or in C6/36 cells) were tested. The titer of inoculum was confirmed by back-titration at the time of inoculation, and stayed within 0.5 log10 difference from the targeted dose. In specific groups, attempts were made to increase the viremia by re-inoculation, either by the subcutaneous or by the intravenous route at 1 dpi. A summary of the experimental groups is presented in Table 1. Using the same mode of inoculation as for the Suffolk breed (group S-A), the 105 PFU dose of Vero E6 produced RVFV in Rideau Arcott cross lambs (group S-B) lead to development of viremia only in three out of four animals at 2 dpi.

Severe maternal complications of preeclampsia warrant delivery. • The adverse conditions These are preeclampsia manifestations that increase the risk of adverse maternal or perinatal outcomes [87] and [95]Table 2 lists the adverse conditions by maternal organ system. Of particular importance are: preterm preeclampsia, chest pain or dyspnoea, or an abnormality of one/more of: oxygen saturation by pulse oximetry, platelet count, serum creatinine, or aspartate transaminase (AST) [87] and [95]. Proteinuria predicts neither

short-term adverse outcomes nor long-term maternal renal prognosis [88] and [89]. Roxadustat HELLP syndrome is represented by its component parts; as we react to HELLP to prevent complications, rather than seeking to avoid its occurrence. How maternal ABT-263 mw adverse conditions may predict fetal or neonatal outcomes in preeclampsia is unclear. The perinatal literature suggests that abnormal fetal monitoring of various types may identify increased fetal risk. Abnormalities in the NST should not be ascribed to antihypertensive therapy [90]. Computerized NST improves perinatal outcomes compared with visual interpretation in high risk pregnancies [91].

Oligohydramnios was not predictive of adverse outcome in observational studies of preterm pre-eclampsia [92]. However, oligohydramnios and abnormalities of Doppler velocimetry of the umbilicial artery have been predictive of stillbirth [86]. The biophysical profile (BPP) has unproven utility Ketanserin in high risk women [67] and [93] and BPP may falsely reassure with early-onset IUGR [94] or preeclampsia [95]. Currently, there is no single fetal monitoring test to accurately predict fetal compromise in women with preeclampsia. Most experts suggest a combination of tests, with emphasis on umbilical artery Doppler when there is IUGR [67] and [96]. Other non-specific risk factors for severe complications of preeclampsia are: immigrant status, young maternal age, nulliparity, lower maternal weight, and in the index pregnancy, multiple pregnancy and early-onset preeclampsia [97].

• What is severe preeclampsia? Definitions vary; most include multi-organ involvement [3], [98], [99] and [100]. We modified our definition of severe preeclampsia to be preeclampsia associated with a severe complication(s). Severe preeclampsia now warrants delivery regardless of gestational age. Our definition excludes heavy proteinuria and HELLP syndrome which is not an absolute indication for delivery. A ‘transient’ hypertensive effect is not associated with an increased risk of adverse outcomes. White coat effect in early pregnancy (∼30%) is common [19]. Forty percent of women progress to persistent hypertension at ⩾20 weeks (i.e., gestational hypertension) and 8% to preeclampsia. Women with ‘white coat’ effect have risks (e.g., severe hypertension, preterm delivery, and NICU admission) intermediate between normotension and either pre-existing or gestational hypertension [15], [60], [66], [101], [102] and [103].

Microbial PAMPs, such as lipopolysaccharides, single-stranded RNA, and bacterial DNA motifs, bind to a family of PRRs called Toll-like receptors (TLR) on innate immune cells and stimulate antigen processing and presentation [16], [17] and [18]. TLRs are widely expressed on dendritic cells (DC) and other professional APCs such as macrophages and B cells. While some TLRs are expressed on the cell surface and act as sensors for extracellular PAMPs (e.g., lipopolysaccharides), a subset of TLR molecules (TLR3, 7, 8 and 9) are expressed

on endosomal membranes and bind selleck products nucleic acid-derived molecules, such as single-stranded RNA of viral origin for TLR7 and 8 [19], [20], [21], [22], [23] and [24] and bacterial unmethylated DNA oligonucleotides (ODNs) containing CpG motifs (CpG ODNs) for TLR9 [14], [25], [26], [27] and [28]. TLR ligands of natural and synthetic origin are potent inducers of innate immune responses and have been shown to effectively stimulate the transition from an innate immune response to an adaptive immune NVP-BGJ398 order response. As such, TLR agonists have been evaluated as potential adjuvants in a variety of applications [4]. To date, only one PRR ligand,

3-O-desacyl-4′-monophosphoryl lipid A (MPL), a TLR4 agonist, has been included as an adjuvant in a FDA- or EMA-licensed vaccine. MPL adsorbed onto alum is utilized in the HPV vaccine Cervarix, licensed in the U.S. and Europe [29], and the hepatitis B vaccine Fendrix, licensed in Europe [30]. Imiquimod, a topically administered TLR7 agonist, has been approved for treatment of genital warts, actinic keratosis, and basal cell carcinoma [31]. Other TLR agonists, such as poly(I:C) (TLR3), imidazoquinolines other than imiquimod (TLR7, 8, or 7/8), and CpG ODNs (TLR9), have failed thus far to enter clinical practice as parenteral adjuvants despite a multitude

of Ketanserin promising data obtained in preclinical and clinical studies [32], [33], [34], [35] and [36]. One of the main reasons for this failure is the delicate balance between the induction of augmented immunogenicity by TLR agonists and safety concerns, which are often related to the generation of systemic inflammatory responses [19], [37], [38] and [39]. Several groups have utilized micro- and nanocarriers, such as virus-like particles, liposomes, and PLGA particles, to encapsulate adjuvants [40], [41] and [42]. Encapsulation of adjuvants reduces systemic exposure of adjuvant and enhances uptake by APCs. Nano-size viruses and particles distribute rapidly to the local draining lymph node where they are taken up by subcapsular macrophages and dendritic cells [41], [43] and [44]. Antigens can also be delivered in particles to target efficient uptake by APCs [36], [41], [45] and [46].

spiralis infected mice. rTs-Hsp70-activated DCs were passively transferred into naive mice three times with intervals of 14

days. The levels of anti-Ts-Hsp70-specific IgG in the sera of these mice were significantly elevated, and these elevations lasted more than 11 weeks without declining ( Fig. 3A). The this website levels of the IgG subtypes were measured, and the results revealed that both IgG1 and IgG2a were induced at similar levels, which indicates that the Ts-Hsp70-activated DCs induced a mixed Th1 and Th2 response in the mice ( Fig. 3B). No anti-Ts-Hsp70 IgG was detected in the mice that received the DCs that were incubated with PBS, the non-relevant protein (Ts-Pmy-N) or LPS. The cytokines IFN-γ, IL-2, IL-4, and IL-6 that were secreted

by the splenocytes that were collected from the mice that were passively transferred with rTs-Hsp70-activated DCs were also measured. The secretions of the Th1 (IFN-γ and IL-2) and Th2 cytokines (IL-4 and IL-6) were significantly elevated in the mice that received the Ts-Hsp70-activated DCs compared those of the groups that received PBS- or non-relevant protein (Ts-Pmy-N)-incubated DCs ( Fig. 4). To determine whether the Ts-Hsp70-activated GDC-0199 price DCs were able to induce protective immunity against T. spiralis infection, the mice that received the DCs were challenged with T. spiralis infective larvae, and the worm burdens were examined at the end of the experiment. The mice that received the rTs-Hsp70-activated DCs exhibited a statistically significant 38.4% reduction in muscle larvae burden compared to the mice that received the PBS-incubated DCs ( Fig. 5). The mice that received recombinant Ts-Pmy-N-incubated DCs did not exhibit a significant reduction in worm burden upon T. spiralis larval challenge.

DCs are central players in the induction and maintenance of immune responses Resminostat and play a prominent role in helminth infections. The infection itself stimulates DC activity, and the infection-induced DC responses are critical for controlling and eliminating the invading agent [26]. In recent years, considerable progress has been made in elucidating the mechanisms behind the interplay between DCs and helminthes [18], [19] and [26]. After interacting with some parasitic helminth antigens, DCs become mature [22], [27] and [28]. The research into the activation and maturation of DCs that are stimulated by helminth antigens has provided a novel approach for the development of vaccines that directly target the antigen-presenting cells [13]. Our previous results indicated that Ts-Hsp70 is a potential vaccine candidate for T. spiralis infection. In the present study, we confirmed that Ts-Hsp70 was able to directly activate mouse bone marrow-derived DCs to mature as characterized by the expressions of typical mature DC cytokines (i.e., IL-1β, IL-6, IL-12p70, and TNF-α) and surface markers (i.e., MHC II, CD40, CD80, and CD86). These results are consistent with the previous observations that T.

The potential of obesity to mitigate breast cancer risk in both premenopausal and postmenopausal patients seems to be

influenced by hormone receptor status; for example, a stronger inverse association between obesity and premenopausal estrogen and progesterone receptor positive (ER +/PR +) breast cancer has been observed compared to ER-/PR- cases [19]. Yang et al. recently found GSK2118436 mouse that obesity was more frequently associated with receptor ER-/PR- breast cancer compared with receptor positive disease in women 50 years old or younger but was more frequent only in patients with PR + postmenopausal breast cancers [22]. An awareness of risk factors for the development of breast cancer in pregnant patients is critical to early diagnosis and treatment of breast cancer. A breast exam should be performed early in pregnancy if possible, and

if exam is performed later in pregnancy, one should exercise vigilance regarding findings. A careful review of chemotherapeutics and their maternal as well as fetal effects should be instituted in a new diagnosis of breast cancer, with close coordination of care among specialists with the patient. No competing financial conflicts exist for any author–investigator. “
“While tuberculosis, especially Selleck BI6727 the pulmonary form is common; tuberculosis of the breast is extremely rare. The incidence of mammary tuberculosis is reported why as less than 0.1% of all breast lesions in developing countries [1] and [2], and diagnosing it is difficult, especially during pregnancy. The signs and symptoms may resemble a malignancy or a non-specific breast abscess, thus labeled a great masquerader (1). We report

a pregnant woman with primary tubercular mastitis who was initially misdiagnosed as having breast abscess. A 31-year-old primigravid pregnant woman was referred to our perinatology unit at 28 weeks of gestation complaining of a painful lump in her right breast that had enlarged progressively over the previous three weeks, as well as new onset pelvic pain. Ultrasonographic examination revealed a single live fetus concordant with 28 weeks, and her pelvic examination revealed minimal cervical dilatation and effacement. A non-stress test revealed regular contractions. The patient was found to have mild fever, and her right breast was minimally enlarged and appeared mildly erythematous when compared to the other side. She had a firm and tender 3–4 cm lump in the upper outer quadrant of the right breast. There was no skin retraction or nipple discharge, and no lymph nodes could be palpated in the axilla or in the cervical region. There was no history of cough or weight loss. The breast ultrasonography revealed a 4 cm complex cystic mass in her right breast. The patient was hospitalized for preterm labor and breast abscess. No family history of breast malignancy was recorded.

Advanced Market Commitments (AMCs) for vaccines are legally-binding agreements to subsidize the purchase, at a given price, of a vaccine that is

not yet available [24]. Efforts to develop an HSV-2 vaccine date back to the 1930s [25]. They received a new momentum in the 1980s, with the emergence of biotechnology, but have so far been unsuccessful PI3K Inhibitor Library [26]. However, several biotech and vaccine companies are investing in the development of an HSV-2 vaccine. Along the same line, there are no vaccines available which effectively protect against a Chlamydia trachomatis genital infection despite many efforts that have been made throughout the years since the 1950s [27]. However, several companies are now in the early phases of clinical trials or considering whether or not they should introduce chlamydia candidate vaccines into their pipeline. As for gonorrhea and trichomonas, interest does not yet seem to have reached this stage. Syphilis was not mentioned during the interviews. Decision to develop a vaccine against STIs is risky as critical scientific information is missing that renders the feasibility and the likelihood of success

of such vaccines uncertain: the mechanisms HKI-272 order of protection are not known; protective antigens have to be identified, and animal models have to be developed or optimized. Moreover, the problem is compounded by the fact that the market for STIs does not seem to warrant the investment inherent in vaccine development. Successful

vaccine development has been based on an understanding of which immunological response is protective. Most successful existing vaccines rely on neutralizing antibodies [28]. Clearly, antibody responses, if necessary, are not sufficient to confer protection to STIs. The problem with HSV-2, chlamydia, gonorrhea and trichomonas is that the immunity induced by natural infection is absent or imperfect. This seriously limits the possibility of defining the types of immune responses that an effective vaccine must include. Ergoloid What is known is that vaccines will have to do better than immunity to natural infection, but which arm of immunity is to be stimulated? There is no viral clearance of HSV-2 infection. The virus persists throughout life in a latent state in the dorsal root ganglia, with episodes of viral reactivation and shedding [29]. Immunity to natural infection by chlamydia, gonorrhea and trichomonas takes time to acquire, is incomplete and of short duration [for reviews, see 1 [30], [31] and [32], in this issue]. Repeat infections are common, and the risk of pathology is known to increase after repeated chlamydial infections [30]. The key question then becomes whether it is possible to design chlamydia vaccines that induce protective immunity without predisposing to more severe pathology.

To guide evidence-based decision making, the advisory group also has recommended national disease burden surveys in children for Hib (2004–2005), rotavirus gastroentritis (2009) and nasopharyngeal carriage of Streptococcus pneumoniae (2009). The agenda for NITAG meetings is adopted by the advisory group in line with the needs of the country or

according to specific proposals from medical universities, MOHME, or WHO. To check details develop technical recommendations and guidelines, the NITAG uses as sources of expert information scientific textbooks, results of local research projects, WHO position statements, and information posted on the websites of WHO, the US Centers for Disease Control and Prevention, and other reputable organizations. In addition, the following criteria

are important for making technical recommendations: the pattern of disease morbidity and mortality in the country, hospitalization rates, disability adjusted life years (DALYs) or quality adjusted life years (QALYs), epidemic potential of the disease, international commitment to disease eradication or elimination, or equity issues. In addition, the NITAG considers economic issues including vaccine cost, overall Epacadostat chemical structure programme costs, results from different economic evaluations (cost-effectiveness, cost-benefit, cost-utility, and others), affordability, and financial sustainability. Whenever the advisory group requires an economic evaluation for its recommendations, the CCDC is asked to conduct an economic survey or study to obtain the relevant information. The advisory group’s recommendations are primarily based on local evidence but regional data also are used if necessary. Recommendations of the advisory group are almost always made by consensus but on rare occasions when members do

not agree, open voting is used to obtain the majority’s decision. When recommendations are finalized, the CCDC is responsible for their dissemination Oxymatrine to the decision makers. Recommendations are then published in a guideline booklet and distributed to public health personnel and medical professionals. The EPI manager and the Director General of CCDC are members of the NITAG and the recommendations are addressed to them. The Director General of CCDC in turn informs the MOHME for implementation of recommendations. Implementation is then considered an obligation since the EPI programme already has government approval. The minutes of meetings are prepared and distributed to the members of the NITAG for their information. The recommendations are also disseminated to the relevant authorities and responsible decision-making bodies for their information and necessary action.

These sequences vary slightly between pseudogenes, for example is more typically LQAEEI to KNRG for msp3 pseudogenes from A. marginale subspecies centrale, but the locations can readily be identified by alignment. Comparing pyrosequencing data to all the known msp2 and msp3 genes showed that all msp2 pseudogenes with the best match in the heterologous strain below 92% variable region identity were detected as absent (−) and all msp3 pseudogenes with below

97% variable region identity were detected as absent (−) ( Table 1). Since the Mosaik alignment parameter −mmp allows a 5% error in aligning reads, we conservatively estimate that variant genes are detected as absent if they have <90% identity, but may not be detected as absent if they have >90% identity. In this study we examined the presence or absence of the pfam01617 buy RAD001 superfamily including genes encoding OMPs 1 through 15, OPAG1-3 and MSP4 [14] and [26]; proteins identified by surface cross-linking including their encoding

genes AM366, 712, 779, 780, 854, 1011, 1051 [15]; and type 4 secretion system genes AM030, 097, 810, 811, 812, 813, 814, 815, 1053, 1312, 1313, 1314, 1315, 1316 [19]. Numbering refers to annotations of the St. Maries, Idaho strain, CP000030. find more To be defined as conserved in A. marginale in Table 4 no segment of the genes was detected as absent in any comparisons of pyrosequenced data from each of 10 U.S. strains of A. marginale with the fully sequenced genomes of Florida and St. Maries, Idaho strains. Pyrosequencing data was previously obtained for A. marginale strains Puerto second Rico, Mississippi and Virginia and in the present study for A. marginale strains Florida, Florida-relapse, Florida-Okeechobee, St. Maries-Idaho, South Idaho, Oklahoma and Washington-O. The average genome coverages were 40×, 12×, 63×, 59×, 76×,

47×, 117×, 37×, 96×, and 108× for the ten strains, respectively, when compared to the completed genome from the Florida strain. Since we did not have current access to the Mississippi strain and coverage was lower for this strain, we also verified that no gene was determined as not conserved solely because of absence in this one strain. The number of high confidence differences between strains (Table 3) was analyzed using Roche/454 gsMapper software to generate the 454HCDiffs.txt file. The base differences and their locations were extracted with the unix grep command and imported into Excel 2008 (Microsoft, Redmond, WA). The number of differences and their respective frequencies (the percentage of different reads versus total reads that fully span the difference location) were tabulated. Finally, for coverage and SNP analyses in Fig. 4 and Table 5, the BAM files generated by Mosaik were processed by samtools version 0.12 to generate pileups. Pileups for genes of interest were extracted to determine coverage for each nucleotide position comparing to both the Florida and St. Maries strains. Final coverages for each gene of interest were graphed using Excel 2008.

À la suite d’une stimulation antigénique, les lymphocytes T CD8+ naïfs Dasatinib in vivo prolifèrent grâce à des molécules de co-activation clé comme en particulier le CD28. Ces lymphocytes T se différencient alors en lymphocytes T cytotoxiques

(qui meurent par apoptose après qu’ils aient accompli leurs fonctions effectrices) et en lymphocytes T mémoires effecteurs ou centraux, qui sont générés en plus petite quantité (5–10 % de la quantité initiale) et dont la fonction est d’assurer une réponse immunitaire plus rapide et plus agressive lors d’une nouvelle rencontre avec l’antigène. Les lymphocytes T CD8+ centraux ont des propriétés d’autorenouvellement. Ainsi, une nouvelle stimulation par les antigènes qu’ils reconnaissent aboutit à la génération de nouveaux lymphocytes T cytotoxiques ainsi qu’à de nouveaux lymphocytes T mémoires centraux et effecteurs. À l’inverse, la stimulation des lymphocytes T mémoires effecteurs aboutit à une prolifération plus modeste avec la mise en jeu rapide des fonctions

effectrices (cytotoxiques ou régulatrices) Selleckchem PLX3397 [15]. Au cours d’une stimulation antigénique persistante au cours du temps, plusieurs de ces cycles d’activation surviennent, aboutissant à des stimulations/proliférations répétées. Dans ce contexte, l’expression du CD28 à la surface des lymphocytes T CD8+ décroît de manière progressive et irréversible, ce qui aboutit à la formation d’une population de lymphocytes T CD8+/CD28− qui possède une capacité de prolifération beaucoup plus faible dans des conditions de culture standards. De manière parallèle, ces lymphocytes acquièrent à leur surface l’expression du CD57 [9], [16] and [17](figures 1B et 2). Ils perdent également progressivement l’expression de l’antigène CD27, traduisant l’état de différenciation avancé de ces lymphocytes. Enfin, ils expriment plus fréquemment l’antigène CD45RA que l’antigène CD45RO et ont

une faible expression de l’antigène CD62L, témoignant bien du caractère « sénescent » de ces lymphocytes [7] and [9]. Ces observations suggèrent ainsi que la population CD28−/CD57+/CD27− dérive de cellules CD28+/CD57−/CD27+. Cette hypothèse est corroborée par la mise en évidence de séquences identiques de la région CDR3 entre ces deux populations lymphocytaires [18]. Les lymphocytes T of CD8+/CD57+ correspondraient donc à des lymphocytes T mémoires/effecteurs activés, dans un état de différenciation terminale ayant le plus souvent perdu leur potentiel cytotoxique et réplicatif et ce, dans un contexte stimulation antigénique chronique [11] and [19]. Ces lymphocytes ont par ailleurs un raccourcissement significatif de la taille des télomères, qui témoigne d’un processus de sénescence tardive [20]. Ainsi, chez le sujet infecté par le VIH, ces lymphocytes produisent de l’interféron-γ ; cependant, en présence de molécules co-stimulatrices, ils se révèlent incapable de s’expandre en réponse aux peptides dont ils sont spécifiques.

The SacB gene driven by RNA-IN promoter was integrated into the chromosome of DH5α, whilst plasmid was incorporated with 150 bp antisense RNA-OUT. In the presence of RNA-OUT antisense regulator, RNA translation of SacB will be silenced and eventually allows plasmid selection in sucrose-containing media [32]. MLN8237 cell line Bacterial strain has been modified to allow suppression of growth essential gene (murA) by repressor protein (tetR) through RNA–RNA antisense reaction [48]. In this system, the plasmid’s replicational inhibitor RNA I could silence the tetR expression.

For this reason, tetR will be turned down and murA expressed for host propagation during the presence of plasmid. The plasmid DNA transcription unit consists of essential components; promoter, intron, signal sequence and polyA, for high expression levels

and targeting of the therapeutic element in the mammalian cells (Fig. 1). Gene promoters contain arrays of regulatory elements to which transcriptional factors bind and interact with each other to regulate transcription. Traditionally, promoters and enhancer regions are derived from pathogenic viruses such as cytomegalovirus (CMV), simian virus 40 (SV40), or murine leukaemia virus. Until now, plasmid DNA promoter from CMV is widely used and has been in clinical trials due to its capability to adapt in an array of tissues and animal models [49]. Unfortunately, a new CMV chimera might be formed by the recombination between CMV promoter from plasmid vaccine and naturally exist wild-type CMV inside the vaccinated person [10]. In fact, INCB024360 in vivo rates of integration or recombination can be influenced by fragments of DNA as short as seven constant base pairs [50]. In conjunction with oncogenesis and mutagenesis risk, highly inter-species-conserved sequences such as housekeeping genes encoding the phosphoglycerate kinase (pgk) and ataxia telangiectasia ATM/E14 should be avoided in promoters and enhancer regions [51] and [52]. Novel synthetic promoters with less risky could be design and selected through bioinformatic tools. Low homology with host sequences could be achieved by using codon optimization software such as OPTIMIZER or gene design software

[53] and [54]. Synthetic promoter also can be generated using ‘fusing technique’. One or two enhancer elements fused to a heterologous promoter sequence. A few investigators no have extended this approach by composing various combination of many regulatory sequences [55] and [56]. For example, Li et al. randomly assembled muscle-specific elements (E-box, MEF-2, TEF-1, and SRE sites) from four different muscle-specific promoters [56]. These novel promoter sequences were screened and one sequence was found having 8-fold higher transcriptional activity comparing to innate muscle promoters. Novel synthetic promoter sequences also can be created by either random ligation of multiple transcription factor binding sites or by DNA shuffling [57].