included both Cariniana micrantha and Carinana decandra ( Procopi

included both Cariniana micrantha and Carinana decandra ( Procopio and Secco, 2008). Other studies of complex genera including Copaifera (Fabaceae, Martins-da-Silva, 2006), Tabebuia (Bignoniaceae, Costa, 2004), and Microphollis (Sapotaceae, Silva, 2004) have found similar mis-identification. Lacerda and Nimmo (2010) reported that at least 43.5% of all species identified after botanical

checking did not appear in the forest inventory and the common practice of matching vernacular check details names to scientific ones proved to be severely deficient. Considering the high importance of correct botanical identification and the uncertainity of forest inventory data which provide the basis for selective logging operations, community and rural extension training in identification is important. Hence, Dendrogene and follow-up projects have provided training course and written guides on this (Ferreira et al., 2004 and Procópio et al., 2005). The Bcl-2 activation Eco-gene model has been used to elucidate genetic processes and the consequences

of logging and forest fragmentation in the long term (Sebben et al., 2008). In this model, data on genetic structure, gene flow and the reproductive biology of Amazonian timber species before and after logging were integrated with data on growth, regeneration and ecology under different scenarios and intensities of logging. The expectation was that these results would help to guide and create new criteria for sustainable logging in the region. Seven species with contrasting ecological and reproductive characteristics were selected for incorporation in the model. The species fit into three ecological groups

(pioneer, climax of fast growth/light demanding and climax Protein kinase N1 of slow growth/shade tolerant categories) and have different reproductive systems (dioecious, monoecious, hermaphrodite), with different pollinators and seed dispersers. The seven species invesitigated were Bagassa guianensis (Moraceae), Carapa guianensis (Meliaceae), Jacaranda copaia (Bignoniaceae), Dipteryx odorata. (Fabaceae), Hymenaea courbaril (Fabaceae), Symphonia globulifera (Clusiaceae) and Manilkara huberi. (Sapotaceae). Dipteryx odorata, J. copaia and M. huberi are hermaphrodites and pollinated by insects, while B. guianensis is dioecious and mainly wind-pollinated, with the participation of trips, a tiny insect. Hymenaea courbaril is hermaphrodite and pollinated by bats, while S. globulifera is hermaphrodite and pollinated by birds, moths and butterflies. Dipteryx odorata and B. guianensis occur at low density in the study area (0.17 and 0.34 individuals per hectare, respectively), while H. courbaril and S. globulifera occur at somewhat higher density (0.58 and 0.88 individuals per hectare, respectively, the latter being for trees >10 cm dbh), and J. copaia, M.

Changes of ±10%

Changes of ±10% Everolimus nmr in the final concentration of master mix and primer pair mix were tolerated by both the PowerPlex® ESI Fast and ESX Fast Systems. An increase of either master mix or primer pair mix to a final concentration of 1.2× had minimal effect on these systems. However, decreasing the concentration of master mix or primer pair mix to 0.8× adversely

affected the signal and balance, particularly for the PowerPlex® ESI Fast Systems (Fig. 1 and Supplemental Fig. 2). Direct amplification is facilitated by the inclusion of AmpSolution™ Reagent in the reaction. Inclusion of AmpSolution™ Reagent in the amplification reaction has no effect on the signal or balance of the profile obtained whether 500 pg of DNA is amplified for 30 cycles or 10 ng for 26 cycles (Supplemental Figs. 3 and 4). No additional amplification artefacts were seen in the presence of AmpSolution™ Reagent over those documented in the technical manuals (data not shown) [14], [15], [16] and [17].

Increasing cycle number from 28 to 30 cycles resulted in the anticipated PCI32765 increase in signal across all loci for the PowerPlex® ESI 17 Fast and ESX Fast 17 Systems. At 32 cycles, the increase in signal was not uniform across all loci, resulting in a locus-to-locus imbalance (Supplemental Fig. 5). Similar results were obtained for the two 16 plexes (data not shown). Increasing cycle number did not result in the appearance of additional artefact peaks in the no-template amplifications reactions (data not shown). We looked at the effect of increasing cycle number from 25 to 27 cycles on blood FTA® cards (Fig. 2) and buccal FTA® cards

(Supplemental Fig. 6), blood on ProteinSaver™ 903® cards (Supplemental Fig. 7), Bode Buccal Collectors (Supplemental Fig. 8), and SwabSolution™ extracts (Supplemental Fig. 9). Signal tended to increase with cycle number for all direct amplification samples. C-X-C chemokine receptor type 7 (CXCR-7) When using two 1.2 mm buccal FTA® punches, full profiles were obtained with all samples at all cycle numbers. Dropout at one locus (in this case one allele at SE33 in one replicate of donor 2) was seen at 25 cycles when using one 1.2 mm buccal FTA® punch (Supplemental Table 3, data not shown for 16 plexes), but not with any of the other direct amplification sample types at any cycle number. The genotypes obtained for a given donor were concordant with each other between cycle numbers and across direct amplification sample types tested. Increasing the annealing temperature to 62 °C from the recommended 60 °C resulted in a significant reduction in signal at amelogenin, D8S1179 and FGA with the PowerPlex® ESI Fast Systems (occasional drop-out at amelogenin and D8S1179 at 62 °C) and dropout occurring at 64 °C along with D2S441 and in some replicates at D2S1338 and D19S433. Overall, 90–100% of alleles were obtainable at 62 °C and 61–76% at 64 °C with the PowerPlex® ESI Fast Systems (Supplemental Fig. 10).

The wet/dry weight ratio was then calculated Brain, heart, liver

The wet/dry weight ratio was then calculated. Brain, heart, liver and kidney were removed, fixed in 4% buffered formaldehyde, and paraffin-embedded. Slices were cut and stained with haematoxylin and eosin. Sections from the regions exhibiting pathologic findings were examined under 400× magnification. A five-point, semiquantitative, severity-based scoring system was used to assess the degree of injury as follows: 0 = normal tissue; 1 = 1–25%; 2 = 26–50%; 3 = 51–75%; and 4 = 76–100% damage out of total tissue examined (Chao et al., 2010). Interferon (IFN)-γ, tumour necrosis factor (TNF)-α and chemokine (C-X-C motif) Selleckchem Alectinib ligand 1 (CXCL1) levels were

quantified. Briefly, the lungs, kidney, liver, brain and heart of control and P. berghei-infected mice were excised and homogenised in cell lysis buffer (20 mM TRIS, 150 mM NaCl, 5 mM KCl, 1% Triton X-100, protease inhibitor cocktail (1:1000, Sigma–Aldrich, USA), and immediately frozen at −80 °C. The total protein content of each tissue homogenate BMN 673 cell line was evaluated by the Bradford method, followed by determination of cytokine production by a standard sandwich ELISA, performed according to manufacturer’s instructions (BD Pharmingen, USA). Plates were read at 490 nm

in an M5 Spectrophotometer (Molecular Devices, USA). Blood–brain barrier (BBB) disruption was evaluated as previously described (Pamplona et al., 2007). Briefly, mice received an intravenous ZD1839 mouse (i.v.) injection of 1% Evans blue (Sigma–Aldrich, São Paulo, Brazil). One hour later, mice were euthanized, and their brains were weighed and placed in formamide (2 ml, 37 °C, 48 h) to extract the Evans blue dye from the brain tissue. Absorbance was measured at 620 nm (Spectramax 190, Molecular Devices, CA, USA). The concentration of Evans blue was calculated using a standard curve. The data are expressed as mg of Evans blue per g of brain tissue. Normality of data was tested using the Kolmogorov–Smirnov test with Lilliefors’ correction,

while the Levene median test was used to evaluate the homogeneity of variances. If both conditions were satisfied, two-way ANOVA followed by Tukey’s test when required was used to compare differences among the groups. Nonparametric data were analysed using ANOVA on ranks followed by Tukey’s test. Parametric data were expressed as means ± SEM, while non-parametric data were expressed as medians (interquartile range). All tests were performed using the SigmaPlot 11 software package (SYSTAT, Chicago, IL, USA), and statistical significance was established as p < 0.05. Mice inoculated with 5 × 106P. berghei-infected erythrocytes demonstrated greater mortality ( Fig. 1A) beginning 6 days post-infection, compared to SAL mice. Parasitemia levels were low at days 1 and 3 post-infection (3.3% and 4.

Moira Elizabeth Schöttler for their assistance in editing the man

Moira Elizabeth Schöttler for their assistance in editing the manuscript. “
“The breathing patterns of patients with chronic obstructive pulmonary disease (COPD) are abnormal, especially in patients

with pulmonary hyperinflation (Aliverti et al., 2004 and McKenzie et al., 2009). Airflow obstructions and mechanical disadvantages of the diaphragm contribute to the changes in the breathing pattern and thoracoabdominal motion observed in these patients (Sackner et al., 1984 and Tobin et al., 1983). Most of these abnormalities suggest a malfunction of respiratory muscles, especially the diaphragm, with the use of sternocleidomastoid (SMM) and abdominal muscle (ABD) being enhanced (Decramer, 1997 and McKenzie et al., 2009).

These patients also exhibit other adaptations, such as modified chest wall and diaphragm shapes, which accommodate the increased volume and adaptations of Alisertib solubility dmso muscles fibers to preserve strength and increase endurance (Loring Baf-A1 purchase et al., 2009 and McKenzie et al., 2009). These abnormalities are associated with poor exercise tolerance, dyspnea and lower functional capacity (Loring et al., 2009). To reduce these consequences, the Joint American College of Chest Physicians/American Association of Cardiovascular and Pulmonary Rehabilitation recommend inspiratory muscle training (IMT) with inspiratory loaded breathing at least 30% of the maximal inspiratory pressure (MIP) (Lotters et al., 2002) as part of rehabilitation programs for patients with COPD (American Association of Cardiovascular and Pulmonary Rehabilitation, 1997). The benefits of IMT have been described by many authors and include increased strength and endurance of the inspiratory muscles, reduced dyspnea and fatigue, increased exercise tolerance and distance walked during the six minute walk test, improved performance in daily activities and an improved quality of life (Geddes et al., 2008, Gosselink et al., Farnesyltransferase 2011 and Shoemaker et al., 2009). Optoelectronic plethysmography (OEP) (Cala et al.,

1996) can be used to elucidate which chest wall (CW) compartment contributes the most to the tidal volume and breathing pattern in different situations. Recent reviews summarized the use of OEP in COPD patients (Parreira et al., 2012 and Romagnoli et al., 2008). Aliverti et al. (2004) found different behavior to increase the tidal volume during exercise: a decrease of end expiratory abdominal volume in euvolemics patients and an increase of end inspiratory abdominal and rib cage volume in hyperinflated patients. Bianchi et al. (2004) also identified during pursed-lip breathing an increased tidal volume associated with increasing end inspiratory rib cage volume and reducing end expiratory rib cage and abdominal volumes. Hostettler et al. (2011) assessed the effect of ILB and identified association between chest wall volume changes and respiratory muscle strength in 12 healthy subjects.

Surveys taken in the reservoir at Lake Oahe (190+ km) have survey

Surveys taken in the reservoir at Lake Oahe (190+ km) have surveys over a shorter time frame (1968–1989). Despite the shorter time frame the trends in reservoir channel change are still considered

applicable. The rate of change in the thalweg bed elevation was calculated as a function of downstream distance and year by determining the minimum elevation of each cross-section (or the maximum depth of the channel), subtracting it from the minimum elevation of the cross-section for the next available year of data, then Osimertinib dividing by the time interval between the two measurements (Eq. (3)). equation(3) BE t1−BE t2t1−t2where BE is the minimum bed elevation (m) and t is time (years). Channels vary naturally through space and time. To attribute a geomorphic change to an anthropogenic disturbance, it must be outside the range of the natural variability and should be statistically significant. This was calculated using the Williams and Wolman (1984) method;

ergodically assuming that longitudinal variation in a single year can approximate ZD1839 cost at-a-station variability through time. The mean pre-dam channel cross-sectional area along the entire segment (irrespective of the defined geomorphic zones) and standard deviation was calculated. The study included all cross sectional data available from 1946, which is the only year of the survey data before the dam was completed. The spatial standard deviation was used to approximate natural variability and compared to the changes at each cross sections. Historical photos from 1950 and 1999 were used to compare change in island area. Photos were georectified using ArcGIS version 10.1. The channel banks and islands were delineated for each year

and the aerial difference between the channel and island boundaries were determined. Water levels along the river vary due to seasonal and annual weather patterns, dam operations, Alectinib concentration tributary influx, and reservoir levels. This consideration is particularly germane with respect to sand bars as the area exposed (and therefore quantified) depends largely on flow depth. The 1999 photo set provides the best comparison to the pre-dam photos (1950) due to similar discharge rates from the Garrison Dam (841 and 835 m3/s respectively or ∼0.7%) and stage gage at Bismarck, ND. All other historical imagery available was collected with discharge differences of 10% or greater related to the pre-dam 1950 images. The spatial extent of the aerial photo analysis ranged from the Garrison Dam to the upper section of Lake Oahe (approximately 130 km downstream of the Garrison Dam); this is the farthest downstream extent of the 1950 images. Image quality of historical aerial photography is often poor, and distortion and clarity are common issues. The aerial photos from 1999 provided by USACE were orthorectified. These orthorectified images were used as a baseline to georectify the 1950 photo set. A minimum of 10 control points per 5 km of river were used.

In their view, however, these impacts are seen as much different

In their view, however, these impacts are seen as much different in scale than those that come later: Preindustrial societies could and did modify coastal and terrestrial ecosystems but they did not have the numbers, social and economic organisation, or technologies needed to equal or dominate the great forces of Nature in magnitude or rate. PCI-32765 order Their impacts remained largely local and transitory, well within

the bounds of the natural variability of the environment (Steffen et al., 2007:615; also see Steffen et al., 2011:846–847). Here, we review archeological and paleoecological evidence for rapid and widespread faunal extinctions after the initial colonization of continental and island landscapes. While the timing and precise mechanisms of extinction (e.g., coincident climate change, overharvesting, invasive species, habitat disruption, BMS-354825 supplier disease, or extraterrestrial impact) still are debated (Haynes, 2009), the global pattern of first human arrival followed by biotic extinctions, that accelerate through time, places humans as a contributing agent to extinction for at least 50,000 years. From the late Pleistocene to the Holocene, moreover, we argue that human contributions to such extinctions and ecological change have continued to accelerate. More than

simply the naming of geologic epochs, defining the level of human involvement in ancient extinctions may have widespread ethical implications for the present and future of conservation biology and restoration ecology (Donlan et al., 2005 and Wolverton, 2010). A growing number of scientists and resource managers accept the premise that humans caused or significantly contributed to late Quaternary extinctions and, we have the moral imperative to restore and rebalance these ecosystems by introducing species closely related to those that became extinct. see more Experiments are already underway in “Pleistocene

parks” in New Zealand, the Netherlands, Saudi Arabia, Latvia, and the Russian Far East (Marris, 2009), and scientists are debating the merits of rewilding North America with Old World analog species (Caro, 2007, Oliveira-Santos and Fernandez, 2010 and Rubenstein et al., 2006). One enduring debate in archeology revolves around the role of anatomically modern humans (AMH, a.k.a. Homo sapiens) in the extinction of large continental, terrestrial mammals (megafauna). As AMH populations spread from their evolutionary homeland in Africa between about 70,000 and 50,000 years ago ( Klein, 2008), worldwide megafauna began a catastrophic decline, with about 90 of 150 genera ( Koch and Barnosky, 2006:216) going extinct by 10,000 cal BP (calendar years before present). A variety of scientists have weighed in on the possible cause(s) of this extinction, citing natural climate and habitat change, human hunting, disease, or a combination of these ( Table 2).

A strong aspect of the study was that the information regarding t

A strong aspect of the study was that the information regarding the wheezing that occurred in the first 12 months of life was obtained when the infants were 12 to 15 months, thereby decreasing the likelihood of recall bias. Other important aspects of this study were the sample size and the use of a standardized questionnaire that allowed for the comparison between the different centers which performed the study. In conclusion, this study demonstrated that some factors were associated with risk of wheezing in

the first year of life, such as family history of asthma, pneumonia, more than six episodes of upper respiratory infections in early life, living in a polluted environment, and use of paracetamol and antibiotics. These factors varied when compared with other Brazilian studies that used the same protocol, which reinforces the need for further studies in other cities NVP-BGJ398 price of Brazil due to the environmental particularities of each region. Prospective follow-ups of wheezing infants are needed to analyze the impact of these risk factors over the long-term in Brazil. The identification of risk factors for wheezing in the first years of life is crucial not only for the diagnosis

of asthma, but also for the development of prevention strategies through public health policies aiming to minimize wheezing morbidity in childhood. Fundação de Amparo à Pesquisa do Estado de Mato p38 MAPK phosphorylation Grosso (FAPEMAT), Process No. 004/2009- 447941/2009. The authors declare no conflicts of interest. The authors would like to thank Fundação de Amparo à Pesquisa do Estado de Mato Grosso (FAPEMAT) for the financial support (Process No. 447941/2009) and the Municipal Health Secretariat of Cuiabá (MT) for the authorization for the use of the public health services

for the performance of this study. “
“The World Health Organization (WHO) defines adolescence as the period from 10 to 19 years, characterized Histone demethylase by intense physical, psychological, and social changes. Rapid growth and nutritional vulnerability also characterize this phase, when there is consolidation of eating habits, which, when adequate, can become a protective factor for obesity, cardiovascular disease, and metabolic disorders in adulthood.1 According to the household budget survey (HBS) conducted by the Brazilian Institute of Geography and Statistics (Instituto Brasileiro de Geografia e Estatística – IBGE) in the years 2008 and 2009, excess body weight was identified in approximately 20% of the adolescent population of the metropolitan areas of Brazil.2 When weight increases due to excess body fat, it can lead to adolescent obesity, which has been considered a predictor of risk for cardiovascular disease, diabetes mellitus, dyslipidemia, and hypertension.

Recent literature reviews suggest that breastfeeding is less comm

Recent literature reviews suggest that breastfeeding is less common among depressed mothers, even though their infants benefit from breastfeeding.25 and 26 Studies from different socio-cultural contexts show almost unequivocally that depressed mothers tend to breastfeed OSI-906 cell line less

or for less time than non-depressed mothers. However, the association between breastfeeding and postpartum depression remains equivocal.25 and 27 When depressed during pregnancy, women are less likely to initiate28 and 29 or to maintain breastfeeding,30, 31 and 32 compared with those with no depressive symptoms. In a recent study on the association between prenatal psychosocial risk factors and breastfeeding intention of Hispanic women, researchers found that women who scored higher in depression at the middle of gestation (about 25.7 weeks) and women who showed persistent depressive symptoms during pregnancy presented a lower intention to breastfeed their babies.33 Other studies have shown that 1/5 of pregnant women are depressed at the third trimester of pregnancy,34 and 35 and that half of these depressed pregnant women will not initiate or breastfeed for three months or more.30 Depression scores at the third trimester were the best predictors of the length of exclusive breastfeeding,

and when considering all the mothers not breastfeeding at three-month postpartum, 37% could be easily detected because of depression during pregnancy.30 Results also EGFR inhibitors cancer showed a significant decrease in depression scores from childbirth to three months

postpartum in women who maintained exclusive breastfeeding for three or more months.30 Exclusive breastfeeding appears to be significantly lower among depressed mothers.36, 37, 38 and 39 Mothers who do not initiate or maintain breastfeeding are more at-risk for depression during the postpartum BCKDHA period.30, 40, 41, 42 and 43 Moreover, when mothers are depressed in the postpartum period, they tend to not initiate28 and 29 or maintain breastfeeding.27, 36, 44, 45, 46, 47, 48, 49 and 50 Some studies have shown that postpartum depression emerges in the sequence of and may result from breastfeeding interruption,40, 41, 42 and 43 suggesting that early cessation of breastfeeding may be involved in the cause of postpartum depression. For example, an association between negative early breastfeeding experiences and depressive symptoms at two months postpartum was found.51 Another study that aimed to assess the association between the infant feeding method and depressive symptoms showed that breastfeeding initiation among multiparous mothers was associated with significantly decreased odds of postpartum depression.43 Other studies suggest that postpartum depression may be involved in the cause of early breastfeeding cessation, and that depressive symptoms have been observed to precede the cessation of breastfeeding.

Antibiotics are the second most often used medication


Antibiotics are the second most often used medication

subgroup in the investigated sample. In several studies,1 and 4 antibiotics appear in the list of the most common drugs used by children, especially those receiving prescribed medications.8 It is known that the main Dinaciclib order respiratory infections are responsible for a large proportion of outpatient antimicrobial prescriptions, demonstrating the intended use of antibiotics.29 Among the most studied antibiotics, amoxicillin was the most often used by children, and similar to results were found by other researchers.8 Amoxicillin is mentioned in international guidelines as first choice of treatment for the most common childhood infections, such as acute otitis media, sore throats, and sinusitis.1 All medications used for the treatment of children must be submitted to the licensing process to ensure the quality, safety, and efficacy in this age group. The four most commonly used medications in the sample (paracetamol, amoxicillin, ibuprofen, and dipyrone) have good documentation for pediatric use, although their indication is not recommended for some age ranges.8, 26 and 27 Moreover, drugs used for the treatment of respiratory tract diseases show little evidence of efficacy, as mentioned previously. One of the most important factors to be considered

BEZ235 in assessing the available evidence in pediatrics is how to deal with the ethical issues of child protection when performing controlled clinical trials. By definition,

these always involve some degree of risk, which in pediatrics should be assumed by parents, based on potential benefits that will not be immediately available to their own children. Despite the Sclareol benefits that the pharmaceutical industry provides in drug development for adults, it is important to emphasize that economic motivation is not prominent for pediatric use. The reduced drug market for the pediatric age range when compared to other ranges, such as adults and the elderly, in addition to the difficulties inherent to performing clinical trials in children, makes the development of these drugs unattractive to the pharmaceutical industry.30 However, as proposed by Coelho et al., the use of a specific list of essential drugs may be part of a comprehensive policy to stimulate the development and manufacturing of medicines for children in Brazil.9 Some limitations of the present study should be considered. The household survey is subject to biases from interviewers and interviewees, which are not always possible to control. The period in which data collection was conducted, April to July of 2013, coincided with the winter season in the region, when there is increased incidence of viral diseases and respiratory infections, which may have contributed to the higher consumption of certain drug classes, such as analgesics, antipyretics, antibiotics, antitussives, expectorants, mucolytics, and antiasthmatic drugs.

Recently, we developed anionic polymer-coated lipoplex of pDNA an

Recently, we developed anionic polymer-coated lipoplex of pDNA and found that CS and PGA coatings for cationic lipoplex produced safe systemic vectors [5]. Anionic polymer-coated lipoplexes have already been developed for pDNA delivery; however, there is little information about the use of the anionic polymer-coated lipoplexes for siRNA delivery. Therefore, in this study, we prepared anionic polymer-coated lipoplexes with CS, PGA and poly-aspartic acid (PAA) and examined the biodistribution and gene silencing

effect in the liver after intravenous C646 injection into mice. 1,2-Dioleoyl-3-trimethylammonium-propane methyl sulfate salt (DOTAP) was obtained from Avanti Polar Lipids Inc. (Alabaster, AL, USA). Poly-l-glutamic acid sodium salt (PGA, 10.5 kDa) was purchased from Sigma-Aldrich Co. (St. Louis, MO, USA). Poly-(α,β)-dl-aspartic acid (PAA, 21 kDa) was obtained from the PolySciTech division of Akina, Inc. (West Lafayette, IN, USA). Cholesterol (Chol) selleck screening library and chondroitin sulfate C sodium salt (CS) were purchased from Wako Pure Chemical Industries, Ltd. (Osaka, Japan). All other chemicals were of the finest grade available. Human breast cancer MCF-7-Luc (TamR-Luc#1) cells stably expressing firefly luciferase (pGL3) were donated by Dr. Kazuhiro Ikeda (Division of Gene Regulation and Signal

Transduction, Research Center Ketotifen for Genomic Medicine, Saitama Medical University, Saitama, Japan) [ 6]. The cells were cultured in Dulbecco’s modified Eagle’s medium

(DMEM), supplemented with 10% heat-inactivated fetal bovine serum (FBS), 100 µg/ml kanamycin and 0.5 mg/ml G418 at 37 °C in a 5% CO2 humidified atmosphere. siRNAs targeting nucleotides of firefly pGL3 luciferase (Luc siRNA), Cy5.5-labeled Luc siRNA (Cy5.5-siRNA), Luc siRNA conjugated with cholesterol (Luc siRNA-Chol), Cy5.5-labeled Luc siRNA conjugated with cholesterol (Cy5.5-siRNA-Chol), nonsilencing siRNA (Cont siRNA) as a negative control for Luc siRNA, Cont siRNA conjugated with cholesterol (Cont siRNA-Chol) as a negative control for Luc siRNA-Chol, cholesterol-modified apolipoprotein B siRNA (ApoB siRNA-Chol) and Cont siRNA-Chol as a negative control for ApoB siRNA-Chol were synthesized by Sigma Genosys (Tokyo, Japan). The siRNA sequences of the Luc siRNA were as follows: sense strand: 5′-GUGGAUUUCGAGUCGUCUUAA-3′, and antisense strand: 5′-AAGACGACUCGAAAUCCACAU-3. In Cy5.5-siRNA and Cy5.5-siRNA-Chol, Cy5.5 dye was conjugated at the 5′-end of the sense strand, and cholesterol was at the 3′-end of the sense strand. The siRNA sequences of the Cont siRNA were as follows: sense strand: 5′-GUACCGCACGUCAUUCGUAUC-3′, and antisense strand: 5′-UACGAAUGACGUGCGGUACGU-3′ [ 7]. In Luc siRNA-Chol and Cont siRNA-Chol, cholesterol was conjugated at the 3′-end of the sense strand.