Input: Claim: Cave paintings offer a glimpse into prehistoric human expression. [SEP] Text: We can easily see the typical form of the human footprint in the prints of 2 and 3 in comparison to the prints produced by contemporary children. This comparison was done in order to prove what seemed to be well understood: that the ancient footprints belonged to human beings. The footprint (Fig. 10. 5a) was made by a female whose left foot was covered by three thin stockings, and thus we recreated a print by a covered foot. Notice the resemblance in the pattern in both prints (Theopetra No. 2 (Fig. 10. 5b) and contemporary female). Fig. 10. 5Contour of the footprint created by covered foot of female, 3 years old (a), and the No 2 footprint of (b). Note the almost similar pattern in the heel and toe regionFull size imageThe footprint (Fig. 10. 6a) was made by a male child (bare foot). Notice the similar pattern (ball, arch and the heel region) with the No 3 footprint of Theopetra (Fig. 10. 6b). Fig. 10. 6Contour of the footprint created by the bare foot; (a) modern child; (b) No 3 footprint of Theopetra Cave. Note the similarity in the pattern of the toes, ball, arch and heel regionsFull size imageAll measurements confirm the first suggestion and leave us with no doubt that they are the footprints of young children (Table 10. 1). Table 10. 1 Measurements of the complete Theopetra footprints and the reference sample of modern Greek children (in mm)Full size tableThis seems highly unlikely. By comparing the length of the feet, we can make the following suggestions. Sex and AgeFootprint No. 2: If a male (young boy) made the footprint, then it should be a child of about 3 years. If, on the other hand, a female made it, it should be between 3 and 4 years of age. Note that we should be particularly careful in our final statements, because a covered foot created the print, and so measurements may not necessarily correspond to the actual dimensions. This means that the print is actually bigger than the foot that created it. 3: this bare footprint was made by a child between 2 and 2. 5 years of age, regardless of sex. The comparison with mean values of Muller et al.

Output: pass


Input: Claim: CRISPR technology allows for precise editing of genomic sequences. [SEP] Text: Additionally, Cre-dependent and constitutive Cas9 expressing transgenic mice were established with EGFP labeling, which provides an animal model for genome-wide targeting and contributes to in vivo investigation [24]. In vivo screens were performed in mouse brain, liver, pancreases, lung and so on. The application of SpCas9 and gRNAs using AAV vectors enabled multiple gene modifications in the adult mouse brain, demonstrating its potential for genetic regulation [33]. Gliomagenesis suppressors were investigated by in vivo stereotaxic injection of AAV carrier sgRNA library in conditional-Cas9 mouse brain [160]. Autochthonous invasion of AAV-mTSGs library in Cre-inducible Cas9 mice liver led to cancer development in situ, and the mice died within 4 months [89]. NIT1 cells (a non-obese-diabetic-derived mouse beta cell line) mutated with GeCKO-v2 were subcutaneously transplanted into type 1 diabetes mouse model to identify genes contributing to autoimmune killing resistance [161]. With the AAV9-LPL gene delivery into the lung, multiple mutations of KRASG12D, p53 and LKB1 were obtained to induce macroscopic tumors. In vivo screening for lung cancer TSGs throughCas9 genome-wide knockout showed that ZNF24 contributed to P65 suppression via NF-κB pathway. Combinational inhibition of -κB, and PD-1 effectively shrank autochthonous KrasG12D/ZNF24−/− lung cancers in mouse [162]. Examination of immunotherapy-treated normal and Tcra-/- mice in vivo by screening showed the loss of CD47 caused resistance to immunotherapy. Deletion of protein tyrosine phosphatase (PTPN2) increased immunotherapy efficacy [88]. screening identified PD-1, Tim-3, and RNA helicase Dhx37 as regulators of tumor infiltration and degranulation. Depletion of Dhx37 improved CD8 T cells efficacy towards triple-negative breast cancer in vivo, and the NF-kB signal pathway was involved in the process [163]. In vivo applications of screening system were summarized in the following Table 3.

Output: corroborate


Input: Claim: Limnology has no relevance for human water usage or conservation efforts. [SEP] Text: The species is reported to exist in two geographically separate subpopulations, as its mopane habitat is bisected by a rocky ridge of miombo (Brachystegia–Julbernardia) woodland which has been speculated to form a barrier to lovebird gene exchange (Dodman1995b; Dodman et al. Reference Dodman, Katanekwa, Apsinwall and Stjernstedt2000). Although this 45-km barrier (Warburton2003) has reduced over the past 30 years and now includes scattered patches of fully grown mopane, there is little surface water within it (Warburton and Warburton and Perrin2005b; CGP 2022, personal observation). The stands of mopane around the Machile, Sichifulo, Ngwezi, and Nanzhila rivers are considered the species’ strongholds (Leonard2005). All the mopane that hosts the Black-cheeked Lovebird is associated with alluvial clay deposits of an ancient wetland, possibly an earlier course of the Kafue River, thus forming an isolated stand (Bingham1994; Leonard2005) and explaining the lovebird’s restricted range (Warburton2003). The southern portion of this range is dominated by seasonal rivers that include the Machile, Sichifulo, Ngwezi, and Simatanga (Dodman1995b; Warburton2003), flowing into the Kasaya River, a major tributary of the Zambezi. The north is dominated by the Nanzhila River, a major tributary of the Kafue, fed by its seasonal tributaries (Ashley and Ashley and Murphy2012). Figure 1. Range and study area of Black-cheeked Lovebird (BCL) Agapornis nigrigenis as documented by Bird(2024). Within the species’ range there are three designations of protected area and one area of informal conservation status. The KNP is Zambia’s largest protected area (Ashley and Ashley and Murphy2012; ZAWA 2004), currently under a co-management agreement with the Zambian government and African Parks. Three game management areas (GMAs) serve as buffer zones between the national park and unprotected land, forming prime legal hunting areas (ZAWA 2010).

Output: contrast


Input: Claim: Cave paintings offer a glimpse into prehistoric human expression. [SEP] Text: (2012) reveals that most probably the two footprints (No 2 and No 3) were made by children aged 3 and 2 years, respectively. Nevertheless, an older age can’t be excluded as a result of volume reduction due to diagenesis. StatureFrom the footprints available, the ones that provide us with more information are No 2 and No 3. The stature for the individual 2 falls in the range 97. 3–106. 5 cm (Robbins 1985, 1986). Applying the percentage of 15% (Topinard 1877), a height of 100. 6 cm was estimated. Finally, the application of the formula of Grivas and his colleagues (2008) gave a stature of 106. 0 cm. The individual that left the bare footprint No 3 probably was a child between 2 and 2. 5 years of age, either male or female. The stature falls in the range 89. 1–97. 5 cm (Robbins 1985, 1986). Applying the percentage of 15% (Topinard 1877), a height of 89. 1 cm was estimated. The application of the formula of Grivas and his colleagues (2008) gave a stature 98. 5 cm. All these calculations are assumptions, because there are several uncertainties when working with footprints. DiscussionFrom the results of our study thus far, we can summarize the following: Footprint size and form: We have limited knowledge about the rate of growth and development of Neanderthal children. Trinkaus (1983) implies that during the first year of their life, Neanderthal children are identical to the children of anatomically modern humans, but this is based mostly on cranial remains. We should also point out that the rate of maturation in Neanderthal children has challenged many scientists, and Dean et al. (1986) proposed that the rate of development in Neanderthal children may have been faster than that in the children of early modern humans. Another recent study of Rosas et al. (2017) notes that Neanderthals’ growth rate is very similar to that of Homo sapiens, in general, but differences have been observed in the development of the brain and spine of these two human groups. These are the main conclusions of a study which focussed on Neanderthal child approximately 8 years old, who lived in the cave of El Sidrón (Spain).

Output: pass


Input: Claim: CRISPR technology allows for precise editing of genomic sequences. [SEP] Text: Table 3 screening in vivo for cancer therapyFull size tableLimitations and prospectionThe advances ofCas technology and screening strategies have revolutionized genetic identification, enabling the dissection of functional genes in specific biological processes and diseases, facilitating drug selection and individualized therapy. screening has demonstrated great potential in cancer therapy by offering methods to combat drug resistance and aggressive behaviors, as well as identifying possible gene targets for novel approaches to treat cancers. However, there are still several obstacles forCas application in clinical cancer treatment, including delivery ofCas9 system, Off-target effect, PAM limitation, as well as multiple gene-editing [117]. In this part, we paid more attention on limitations of screening system and CAR-T cell therapy for cancers. Limitations of screening systemscreening delivery primarily relies on lentiviral and AAV vectors, which are crucial tools for either ex vivo or in vivo investigation. Of which, AAV vector has the advantages with mildly immunogenic and long-term transgene expression in post-mitotic cells, making it a leading platform for in vivo cancer therapy [164]. However, AAV vector showed some drawbacks in manufacturing, packaging size limitation, vector quality control and editing specificity, as described in the recent review [118]. Except the delivery limitations, the occurrence of off-target effects and unintended mutations induced by technology are barriers to its application in clinical therapy. SpCas9 protein showed the ability to identify PAM sequence and cut specific DNA region in the system. Due to the tolerance of gRNA recognition and nucleotide indels in the target region, even a single guide can generate thousands of off-targets as detected by sensitive high-throughput sequencing methods such as GUIDE-Seq and -seq [138, 143]. This raises concerns regarding the application of technology in gene therapy [165]. The reason of the off-target effect is the conformational states of HNH domain. The activated conformation of HNH increases DNA cleavage efficiency for DNA double-strand break formation, leading to both on- and off-target effects [166]. To minimize the probability of off-target mutagenesis, other high-fidelity nucleases such as SpCas9-HF1, eSpCas9 and HypaCas9 were developed [167, 168].

Output: corroborate


Input: Claim: Limnology has no relevance for human water usage or conservation efforts. [SEP] Text: Fifteen forest reserves are intended to conserve the headwaters of major streams and rivers, although these are largely encroached by both settlements and agricultural fields (Shitima2005). Informal conservation status is bestowed by Bird and (IBA), notable as a stronghold of the Black-cheeked Lovebird (Leonard2005). No settlements are present in the national park, but they exist in the park’s GMAs (Leonard2005). The human population within the species’ range is estimated to be about 55,000, with the majority being semi-nomadic cattle-herders, fishermen, and small-scale crop farmers (2015). For the purpose of this study, the species’ range is divided into a northern and southern region based on area management type, with the north falling within state-protected areas under the Act of 2015 and the south forming predominantly communal or customary owned land. Pool location and monitoring Data were collected over a period of 30 months; pool monitoring was undertaken between December 2018 and December 2020, while searches for other pools continued for an extra six months (May–October 2021) to ensure wider coverage of the species’ range and to compensate for fieldwork lost to COVID-19 travel restrictions on CGP between May and December 2020 (pool monitoring by community members – see below – continued in this period). Transects covering multiple habitat types including mopane woodland were undertaken to survey the lovebirds using distance sampling (Buckland et al. and Colton2005). An initial 70 transects were pre-selected in ArcGIS Pro using the tool (ESRI 2019). To locate pools, a combination of methods was used. A desk review collected the coordinates of all the pools used by lovebirds during earlier studies (Warburton2003). Pools were also located by interviewing local inhabitants, and along transects. Transects were run systematically across the study area from east to west but, to increase the likelihood of finding pools, some transects followed footpaths and streams (Mirzaei and Mirzaei and Bonyad2016). Along transects we constantly attempted to record lovebird presence and numbers at pools.

Output: contrast


Input: Claim: Cave paintings offer a glimpse into prehistoric human expression. [SEP] Text: Foot function and footwear: Neanderthals used fire; they certainly buried their dead; they seem to have self-medicated with local plants; and they undoubtedly used foot coverings. It is very crucial to know whether the Neanderthals and early humans have the same foot function. and his colleagues (2016) conclude that foot function has remained almost unchanged, perhaps experiencing evolutionary homeostasis, for the last 3. 66 million years. The archaeological record has limited evidence of footwear. The most ancient evidence appears to be in Theopetra Cave. The footwear probably had significant use among humans, who may have had various forms of foot covering, to provide insulation and protection from cold weather and rough substrate (Trinkaus 2005). This phenomenon of footwear use must have been widespread although the archaeological findings are very rare. Age: Children between 2 and 4 years of age produced the footprints, bearing in mind that these children could belong to early Homo sapiens or Neanderthals. The analysis of the footprints gave a clear reconstruction of the facts that occurred at this time in the cave. Firstly, the cave was in use during the (Kyparissi-Apostolika 1999; Karkanas 2001; Valladas et al. 2007; Facorellis et al. 2013). There are traces of fire remnants on an ashy wet surface, and the footprints are very near these remnants. This could mean that the children were walking and playing in the area surrounding the hearth. We suppose that there are several trails (at least two) which were made by different individuals. The evidence that an individual who wore some kind of foot covering made the first two prints supports this. A bare foot has made the third print, whereas the fourth one is not clearly distinguished, either because it has been rotated or because other footprints have been made on it. The study of the footprints reconstructs and brings the children in our eyes: We can imagine these children playing in the cave and leaving their traces in the ashy wet surface around the burnt remnants. Neanderthal children or not? Through dating of specimens from the layer on which the footprints were found, it became obvious that at the specific time point early Homo sapiens and Neanderthals coexisted. The former make an appearance in Europe early, at about ~210 ka in Apidima Cave, Greece (Harvati et al. 2019). The latter resided in Europe, and their remains are found all over the continent.

Output: pass


Input: Claim: CRISPR technology allows for precise editing of genomic sequences. [SEP] Text: In addition, PAM sequence limitation for Cas9 has been broadened by the identification of KKH SaCas9 variant, which exhibits robust genome editing activities with the PAM () while maintaining comparable levels of off-target effects [169]. Anti-is another obstacle to overcome because of the restriction of targeting specificity and activities. The VI-inhibitors acrVIA1-7 from phage exhibit the ability to block Cas13a RNA targeting and dCas13a-mediated single nucleic acid editing. Specifically, AcrVIA1, 4, 5 and 6 bind to LwaCas13a, while AcrVIA2 and 3 interact with LwaCas13-crRNA complex [170]. Limitations of CAR-T cancer therapyAlthough CAR-T showed success of B-cell malignance treatment, its usage in solid tumors still have some limitations such as T-cell exhaustion, lack of CAR-T cell persistence, and cytokine-related toxicities. To address these challenges, technology has been used to generate safe and potent allogeneic universal CAR-T cell products for cancer immunotherapy [152]. However, hurdles remain for solid tumor CAR-T therapy due to target antigen heterogeneity, unable to pass through vascular endothelium to target tumor cells, and the immunosuppressive tumor microenvironments [171]. As viral vectors are commonly used for delivering CAR-T cells, safety concerns have arisen. To address this issue, virus-free -CAR (VFC-CAR) T cells were generated [172]. Virus-free CAR-T cells (PD1-19bbz) were generated and a clinical trial was performed and registered at www. clinicaltrials. gov (NCT04213469) [173]. Future perspectivesGiven the capacity of to precisely modify the human genome in cells, ethical considerations have emerged as a pivotal factor for its application in genetic manipulation [174,175,176]. The challenges posed by off-target effects and unintended mutations serve as barriers to the clinical implementation of technology. However, extensive efforts have been made to mitigate these concerns through the development of novel strategies, rendering technologies indispensable tools for elucidating gene functions and noncoding elements involved in tumorigenesis, as well as facilitating the creation of next-generation cancer immunotherapies. In summary,Cas system continues to play an essential role in advancing human cancer research and clinical therapy.

Output: corroborate


Input: Claim: Limnology has no relevance for human water usage or conservation efforts. [SEP] Text: Distribution of pools across the study area; the red border represents the northern region while the black line represents the south. Overall patterns of pool usage There were significant differences in the proportions of pools used across the four pool types (χ² = 11. 8, df = 3, P = 0. 008) (Table 3). From the GLM results, used pools tended to be close to mopane, have a high basal area of surrounding trees, and lack human activity (Table 4). In the dry season, however, pool usage was influenced by distance to mopane, the basal area of surrounding trees, human activity, distance to nearest pool, and pool size, while pool type was not significant (Table 4). In the wet season, key parameters influencing lovebird usage of pools were almost the same but with pool type replacing human activity. Table 3. Overview of characteristics of used and unused pools Table 4. Most supported models from logistic regression for overall and seasonal usage, and numbers of Black-cheeked nigrigenis using pools from Poisson regression. Figures given are coefficients for each predictor. *P <0. 05; **P <0. 01; ***P <0. 001; ns = not significant AP = artificial pool, MP = mopane pool, RP = river pool (reference level used = artificial pool). No activity present = reference level on human activity Variation in lovebird numbers at pools Numbers of birds were influenced only by pool type and were higher where human activity was lower. Pool monitors paid a total of 837 visits to used pools, of which 412 were in the morning. Time of day (morning or afternoon) did not significantly affect the number of birds recorded (χ² = 0. 75, df = 1, P = 0. 39). Lovebird numbers at monitored pools varied with pool type and season (wet/dry). The correlation between season and pool type in the dry season (rₛ = −0. 73, n = 53, P <0. 001) was stronger than that in the wet season (rₛ = −0. 48, n = 53, P = 0. 01), suggesting that pool type has a more substantial and significant impact on the number of birds visiting during the dry season compared with the wet season (an average of 1,920 in the dry season and 2,064 in the wet season).

Output: contrast


Input: Claim: Cave paintings offer a glimpse into prehistoric human expression. [SEP] Text: Evaluating all the available information, it is difficult to conclude what kind of individuals left these footprints. How do we know that they are Neanderthal children? We do not know this because the form and shape of the footprints are not consistent with the known anatomy of Neanderthals, from various other sites. Duveau et al. (2019: 19411) note, “They are relatively broader, especially in the midfoot, than the footprints made by Homo sapiens, which corresponds to a more robust foot and a less pronounced arch”. The fact that the footprints of Theopetra have been made by children aged 2–4 years would lead one to hypothesize that probably the foot at this age is not fully developed. On the other hand, the cultural findings (lithic artefacts) seem to be Mousterian (the typical technological expression of Neanderthals) and have led to the conclusion that the footprints were made by Neanderthal children. ConclusionBoth the palaeoanthropological and archaeological records suggest that foot covering was present during the Middle Palaeolithic. The only evidence that helps us in this case for establishing chronology is the lithic material found in this layer. If eventually the assessment that the tools are Mousterian (Panagopoulou 1999, 2000) is confirmed, we will positively assume that they were Neanderthal children. But even if they turn out to be the children of early Homo sapiens, the significance of the findings is still great, since that would prove beyond reasonable doubt the presence of anatomically modern humans in Europe at approximately ~135 ka much earlier of what was thought until now. 13. 189. 529. 223. 29. 51. 804. 309. 9414. 189. 1048. 1442. 95. 2020. Harvati, K. , Röding, C. , Bosman, A. M. , Karakostis, F. A. , Grün, R. , Stringer, C. , Karkanas, P. , Thompson, N. C. , Koutoulidis, V. , Moulopoulos, L. A. , Gorgoulis, V. G. , & Kouloukoussa, M. (2019). fossils provide earliest evidence of Homo sapiens in Eurasia. Nature, 571, 500–504. 399. 1997. ERAUL 85 (125. 394.

Output: pass


Input: Claim: CRISPR technology allows for precise editing of genomic sequences. [SEP] Text: AbstractThe determination of null- or low-expressed HLA alleles is clinically relevant in both hematopoietic stem cell transplantation and solid organ transplantation. We studied the expression level of a questionable (Q) HLA-B*38:68Q allele, which carries a 9-nucleotide (nt) deletion at codon 230–232 in exon 4 of HLA-B*38:01:01:01 usingCas9 gene editing technology. /Cas9 gene editing of HLA-B*38:01:01:01 homozygous EBV B cell line resulted in one HLA-B*38:68Q/B*38:01:01:01 heterozygous and one HLA-B*38:68Q homozygous clone. Flow cytometric analysis of monoclonal anti-Bw4 antibody showed the protein expression of HLA-B*38:01:01:01 in homozygous cells was 2. 2 fold higher than HLA-B*38:68Q/B*38:01:01:01 heterozygous cells, and the expression of HLA-B*38:68Q/B*38:01:01:01 heterozygous cells was over 2. 0 fold higher than HLA-B*38:68Q homozygous cells. The HLA-B*38:68Q expression was further confirmed using anti-B38 polyclonal antibody. Similarly, the expression of the HLA-B*38:01:01:01 homozygous cells was 1. 5 fold higher than that of HLA-B*38:68Q/B*38:01:01:01 heterozygous cells, and the HLA-B*38:68Q/B*38:01:01:01 heterozygous cells was over 1. 6 fold higher than that of HLA-B*38:68Q homozygous cells. The treatment of HLA-B*38:68Q homozygous cells with IFN-γ significantly increased its expression. In conclusion, we demonstrate that HLA-B*38:68Q is a low-expressing HLA allele. /Cas9 technology is a useful tool to induce precise gene editing in HLA genes to enable the characterization of HLA gene variants on expression and function.

Output: corroborate


Input: Claim: Limnology has no relevance for human water usage or conservation efforts. [SEP] Text: This aligns with evidence across south-western Zambia where reductions in rainfall have been observed since 1978 and are projected to continue past the year 2050 (Libanda and Libanda and Ngonga2018; Musonda et al. Reference Musonda, Jing, Nyasulu and Mumo2021). Overall, the results of our study provide evidence of additional conditions that need to be met to ensure the long-term survival of Zambia’s endemic arid-country parrot. The probability of a pool being used by Black-cheeked Lovebirds increased with greater proximity to mopane, a larger basal area of adjacent trees, and an absence of human activity. Being dependent on mopane for nesting and roosting cavities as well as general cover, the species presumably saves energy and reduces predation risk by foraging and drinking close to mopane (Devereux et al. Reference Devereux, Whittingham, Fernández-Juricic, Vickery and Krebs2005; Molokwu et al. Reference Molokwu, Nilsson, Ottosson and Olsson2010; Mzumara et al. Reference Mzumara, Martin, Tripathi, Phiri and Amar2019). Visits to distant and isolated sites by other bird species inhabiting arid areas, for example in the dry season when water is scarce, are only undertaken in large numbers that may lower the individual chances of predation (Devereux et al. Reference Devereux, Whittingham, Fernández-Juricic, Vickery and Krebs2005; Lima et al. and Dill1990). Variations in numbers drinking at pools probably reflected variations in the lovebird’s abundance within its range (Warburton and Warburton and Perrin2005b; Phiri et al. in prep. ). Arid-habitat species minimise their energetic expenditure to maintain a water balance (Jong1976; Smit and McSmit and McKechnie2015), so using pools close to mopane where the species roosts and nests (Warburton and Warburton and Perrin2005a) must prevent large energy outflows (Bryant1997). At pools the lovebirds initially gather in nearby bushes or trees before descending collectively to drink (Dodman et al.

Output: contrast


Input: Claim: Cave paintings offer a glimpse into prehistoric human expression. [SEP] Text: -19 PreprintsProtocolsVideosJournalsTools & a 10. 21203/rs. 3. rs-927119/v1This work is licensed under a CC BY 4. 0 LicenseStatus:PostedVersion 1posted You are reading this latest preprint versionAbstractinvolves the accumulation of oxidized low-density lipoprotein (OxLDL)-laden foam cells (FC) within the blood vessel intima. Macrophages are transformed to FCs by the uptake of OxLDL, a process mediated by scavenger receptors (SRs), which include MSR1, CD36 and CD68. Atherosclerosis differs in prevalence across ethnic groups, being less common in African than Indian or European populations. Therefore, our aim was to measure SRs gene expression from macrophages isolated from these three ethnic groups. participants were recruited from each ethnic group (African, European and Indian). Anthropometry and fasting serum lipid and glucose levels were measured. Monocytes were isolated from whole blood and converted to macrophages using standard cell culture procedures. Macrophage RNA was isolated, and reverse transcribed to cDNA. Relative gene expression was calculated using the ∆∆Ct relative quantification method with β-actin used as the normalization control. Multivariable regression analysis was performed to identify the determinants of macrophage SRs expression. of the CD36 gene correlated with age (β = 0. 33, p = 0. 02), LDL (β = 0. 29, p = 0. 03) and CD68 expression (β = 0. 57, p < 0. 001). Expression of CD68 correlated with triglycerides (β = 0. 45, p = 0. 005) and European ethnicity (β = 0. 56, p < 0. 001). Expression levels of MSR1 correlated positively with LDL (β = 0. 27, p = 0. 01) and negatively with HDL (β=-0. 33, p = 0. 003) and African ethnicity (β=-0. 73, p < 0. 001). data suggest that serum lipids may modulate foam cell formation via effects on macrophage SRs gene expression. Furthermore, ethnic differences in atherosclerotic plaque formation may be mediated through differential CD68 and MSR1 expression levels within macrophages.

Output: pass


Input: Claim: CRISPR technology allows for precise editing of genomic sequences. [SEP] Text: 9 reported a patient, who carried HLA-A*30:14L and HLA-A*02:01 alleles, was mistyped as HLA-A*02:01 homozygous. The mistyping of HLA-A*30:14L led to severe GVHD in the patient transplanted with HSCT from an HLA-A*02:01 homozygous donor. Currently, there are 663 null alleles (N), 44 questionable alleles (Q), 5 low expression alleles (L), and 1 soluble alleles (S) identified in IMGT/HLA data base2. The HLA matching of the questionable alleles between the donor and recipient remains challenging. Therefore, it is important to determine the expression patterns of abnormally expressed HLA variants7. (clustered regularly interspaced short palindromic repeats) is an adoptive immune system in bacteria that protects the bacterium from invading foreign genetic elements such as plasmid and bacteriophages10. /Cas9 system contains two components: a guide RNA (gRNA) and a -associated endonuclease (Cas protein)11,12. The gRNA is a short RNA composed of a scaffold sequence needed for Cas-binding and a user-defined ∼20 nucleotide spacer that defines the genomic target to be modified13. The gRNA spacer sequence could be designed to target DNA sites with Motif (PAM)14,15. The most common PAM sequence recognized by Cas9 is NGG that is found directly downstream of the target DNA. /Cas9 cuts double strand DNA to generate double strand breaks (DSBs) between 3–4 bp upstream of the under the guidance of gRNA16. The DSBs can be repaired by non-homologous end joining (NHEJ), which is an error-prone process that introduces unpredictable insertions and deletions (indels); DSBs can also be repaired by homology directed repair (HDR) with the presence of DNA template, which induces desired DNA editing11,12,17.

Output: corroborate


Input: Claim: Limnology has no relevance for human water usage or conservation efforts. [SEP] Text: Reference Dodman, Katanekwa, Apsinwall and Stjernstedt2000; Warburton and Warburton and Perrin2005b). They avoid pools that lack perches within 20 m of the water’s edge (Warburton and Warburton and Perrin2005b; CGP 2021, personal observation), and in our study basal area of nearest trees served as a proxy for the quantity of available perches and cover near pools (Brumelis et al. 2020). Nevertheless, although our results confirm the strong association between lovebirds and mopane, some used pools were surrounded not by pure mopane but by mixtures of mopane and other tree species or by other species alone, including Combretum imberbe and Diospyros mespiliformis (Dodman et al. Reference Dodman, Katanekwa, Apsinwall and Stjernstedt2000; CGP 2019, personal observation). Overall, fringing vegetation at water points is known to be more critical for small and mid-sized forest birds, whose vulnerability to predation increases when they access drinking sites with too distant or no vegetation (Votto et al. Reference Votto, Schlesinger, Dyer, Caron and Davis2022). Lovebirds are extremely cautious drinkers that avoid drinking from a pool during periods of loud and obvious human activity (Mzumara et al. Reference Mzumara, Perrin and Downs2016; Warburton and Warburton and Perrin2005b; CGP 2021, personal observation). In the flood months (February–April) water is abundant in most of the shallow wells within villages and livestock enclosures (Lewanika, Bernard, Mosepele and Ramberg2003; Thole and Thole and Dodman1997), reducing the need for human activity at natural pools. In the dry season, when smaller pools dry out, most pools, particularly the larger ones, saw an increase in human activity, coinciding with a steady increase in the number of lovebirds visiting river and artificial pools. Other factors known to influence the numbers of lovebirds using pools include local lovebird abundance, pool physical characteristics, and breeding regime (Mzumara et al. Reference Mzumara, Perrin and Downs2016; Warburton and Warburton and Perrin2005b, and Perrin2005d).

Output: contrast


Input: Claim: Cave paintings offer a glimpse into prehistoric human expression. [SEP] Text: PathologyAtherosclerosisFoam cellScavenger receptorsMacrophagesFiguresFigure 1BackgroundThe major cause of heart disease and myocardial infarction in western society is atherosclerosis (1). Atherosclerosis involves the accumulation of oxidized low-density lipoprotein (OxLDL)-laden foam cells (FC) within the blood vessel intima (1). Foam cell formation is the principal event in the development of atherosclerotic plaques. Foams cells form from macrophages following macrophage uptake of oxidised-LDL (oxLDL) (2). The uptake of oxLDL is accomplished by specific receptors on the macrophage cell surface, the scavenger receptors (SRs). Scavenger receptors are a ‘superfamily’ of membrane-bound receptors that bind and internalize modified LDL. Identification of these SRs in macrophages was first described in the 1970s by Brown and colleagues (3). There are 3 main sub-types of SRs: macrophage scavenger receptor 1 (MSR1), CD36 and CD68 (2). The CD36 SR is an 88-kDa transmembrane glycoprotein receptor that belongs to the class B SR family (4). It is encoded by the CD36 gene, located on chromosome 7 (5, 6). This gene is expressed on various cell types including monocytes and macrophages and its expression levels are increased by a fat-rich diet, inflammation and oxidative stress (5, 6). The MSR1 protein is a 77-kDa cell surface glycoprotein and is a member of the class A SR family (7). It is highly expressed in macrophages but is also present on vascular smooth muscle and endothelial tissues and it mediates the uptake of oxLDL into macrophages (8). The CD68 molecule is a 110-kDa glycoprotein that is a member of the class D SR family (9). The CD68 gene is located on human chromosome 17 and it is highly expressed by monocytes and tissue macrophages (10, 11). Macrophages develop from monocytes in response to specific immunological stimuli, including macrophage-colony stimulating factor (M-CSF) (12).

Output: pass


Input: Claim: CRISPR technology allows for precise editing of genomic sequences. [SEP] Text: Two types of the DNA template can be used for HDR: a small single stranded DNA (ssDNA) oligonucleotide with 30–67 nt homology arms flanking the gene editing site18 or a double stranded DNA (dsDNA) plasmid with long homology arms of 1–3 kb19. The recent discovery ofCas9 system provides a faster and more economical approach to gene editing20 compared to the traditional zinc-finger nucleases (ZFNs)21 and transcription activator-like effector nuclease (TALEN) methods22. The goal of this study was to generate homozygous and heterozygous cells carrying the HLA-B*38:68Q with deletion at codon 230–232 at exon 4 usingCas9 gene editing to study the effect of this mutation on HLA-B*38:01:01:01 expression. ResultscrRNA design and selectionWe identified a new HLA B allele that is similar to HLA-B*38:01:01:01 except for a nine-nucleotide deletion (5′--3′) at codon 230 to 232 that results in a coding shift at α3 domain of HLA-B38 (Fig. 1A). The sequence was submitted to the GenBank database (accession number MF069211) and IMGT/HLA databases (submission number HWS10028807). Since the expression level of this novel B*38 allele is unknown, it was named HLA-B*38:68Q. To determine if the deletion at codon 230–232 affected the level of protein expression, the homozygous HLA-B*38:01:01:01 EBV B cell line TEM665 was used to generate homozygous HLA-B*38:68Q alleles to study its expression (Fig. 1B). and was used to design crRNAs targeting the DNA sequences close to the 9-nt deletion site at exon 4 of HLA-B*38:01:01:01 (Fig. 1C). crRNA1 (5′--3′) was designed to recognize −12 to −31 bp upstream of the 9-nt deletion, and crRNA2 (5′--3′) was designed to recognize −2 to +9 bp sequence of the 9-nt deletion.

Output: corroborate


Input: Claim: Limnology has no relevance for human water usage or conservation efforts. [SEP] Text: In density surface models developed for the species (Phiri et al. in prep), mopane cover was a significant determinant of local abundance, which in turn will likely influence bird usage of pools. Our study also confirms that the physical features of pools play a significant role in determining their usage by lovebirds, aligning with previous findings related to desert birds (Fisher et al. Reference Fisher, Lindgren and Dawson1972). Mopane and grassland pools dried out by October, the former refilling after a heavy downpour, the latter refilling according to the flooding regimes of the adjacent river (Naidoo et al. and Du Preez2020). Consequently, these two pool types showed the most variation in the number of birds across the monitoring period. By contrast river pools, which form in major channels along seasonally dry riverbeds (Warburton and Warburton and Perrin2005b) and last until the onset of the rains, generally hosted a constant number of birds, with a slight increase in the dry season. Similarly, artificial pools recorded the highest number of birds between October and December, when water is extremely scarce in the study area. Greater dependence on artificial pools has been reported previously in the lovebirds’ southern rather than in their northern subpopulation (Warburton and Perrin 2000b). The north is dominated by the and its inundated floodplains (Ashley and Ashley and Murphy2012), which have water all year round. In the south, locals dig small dams or maintain dams left after road maintenance activities for their livestock and gardens; we found that nearly 40% of all artificial pools located in the south were used by lovebirds. River pools and artificial pools evidently play a pivotal role in maintaining lovebird populations throughout the annual cycle. Potential management interventions to improve the long-term survival prospects of the Black-cheeked Lovebird include simply enhancing the capacity of artificial pools to meet the needs of the lovebirds, by ensuring that each pool’s characteristics meet the criteria of the best-fitting model. However, the creation of small, shallow-sided, undisturbed pools in or near mopane woodland with larger trees within 50 m must also be considered. This highlights the wider need to review existing best practices of water resource provisioning for wildlife species, to devise interventions that will extend the water retention of mopane pools beyond the wet season.

Output: contrast


Input: Claim: Cave paintings offer a glimpse into prehistoric human expression. [SEP] Text: This molecule is the main facilitator of the differentiation of monocytes into macrophages in vitro and its presence has been demonstrated in atherosclerotic lesions (13). The risk of atherosclerosis is modified by a number of factors and disease states. Thus, increasing age, high low density lipoprotein- cholesterol (LDL-C), low high density lipoprotein- cholesterol (HDL-C) and high glucose levels, increased body mass index (BMI) and low socioeconomic status are all associated with an increased risk of atherosclerotic disease (14). Atherosclerosis differs in prevalence across ethnic groups, being less common in African than Indian or European populations (15). Furthermore, in Norman and colleagues found that cholesterol-attributable mortality is highest in Indian subjects (22. 2%), followed by European (20. 5%) and mixed-ancestry (8. 8%) population groups. Cholesterol attributable mortality was lowest in the black African group, at only 1. 8% (16). The reasons for these ethnic variations in the prevalence of atherosclerosis and the contribution of cholesterol to mortality are not fully understood. Given that factors associated with a higher risk for atherosclerosis can modulate SR gene expression, we hypothesised that ethnic differences in the level of atherosclerotic disease may be related to differential macrophage expression of the SRs across ethnic groups. The aims of the current study were therefore to measure SR gene expression from macrophages isolated from African, European and Indian populations to determine if SR expression levels mirror the reported ethnic differences in the prevalence of atherosclerosis. We also sort to determine whether anthropometric, demographic and cardiometabolic factors influence macrophage SR gene expression. recruitment Ten healthy participants per ethnic group (African, European, Indian) aged ≥ 40 years were recruited to participate in the study from the greater Johannesburg area by convenience sampling. The three groups were matched for gender. Anthropometric measurements (weight, height, and hip and waist circumferences) were collected for all participants and blood drawn through venous sampling. Body mass index (BMI) was calculated using weight and height (BMI = weight (kg)/ height2 (m)). The participant’s ethnicity was self-reported. Participants receiving therapy for high lipid levels, participants with diabetes or those with a history of cardiovascular problems were excluded from the study.

Output: pass


Input: Claim: CRISPR technology allows for precise editing of genomic sequences. [SEP] Text: Both crRNAs share a ssDNA target with a 67-nt of left arm and a 30-nt of right arm of the deletion site (Fig. 1C). The crRNA1 and crRNA2 were then mixed with universal tracrRNA to form gRNA1 and gRNA2. Next, Cas9 protein (1. 5 µg) and gRNA1/gRNA2 (360 ng) were mixed to form Cas9 ribonucleoprotein (RNP), respectively18. Twenty four electroporation conditions were tested to optimize transportation efficiency using Neon transfection system23 (1). The program of the highest transportation efficiency (58. 4%) was selected for transfecting HLA-B*38:01:01:01 homozygous EBV B cell line. Our results showed that gRNA1 induced 22. 6% DSB cleavage and gRNA2 induced 13. 8% DSB cleavage using GeneArt® Detection assay. The gRNA1 was chosen for transfection with ssDNA target due to its high efficiency. Figure 1crRNA and ssDNA target design. (A) Comparison of HLA-B*38:01:01:01 and HLA-B*38:68Q in nucleotide and protein sequences. Yellow box highlights the 9-nt deletion mutation and “. ” indicates the deletion. Numbers above sequence correspond to amino acid codon positions. (B) HLA typing of TEM665 homozygous HLA-B*38:01:01:01 EBV transformed B cell line used for gene editing. (C) Design of crRNA and ssDNA target on HLA-B*38:01:01:01. crRNA1 and crRNA2 are colored in green. (PAM) recognition sites are marked in blue. The red box indicates the location of 9-nt deletion. “. ” means that the ssDNA oligo contains the 9-nt deletion. Blue arrows show the position of double strand breaks (DSBs) that should be introduced byCas9. Full size imageGeneration of HLA-B*38:68Q clones usingCas9 gene editingGene editing of HLA-B*38:01:01:01 homozygous EBV B cell line byCas9 using gRNA1 resulted in 25 single cell derived clones.

Output: corroborate


Input: Claim: Limnology has no relevance for human water usage or conservation efforts. [SEP] Text: Water provision programmes are prominent in many key biodiversity areas in southern Africa in response to increases in water stress (Selebatso et al. Reference Selebatso, Maude and Fynn2018). High-profile sites with such programmes include the Game Reserve, and Hwange and Parks, where diesel engines are used to pump water into pools, evidently promoting the presence and abundance of certain bird species, particularly seedeaters (Abdu et al. Reference Abdu, Lee and Cunningham2018; Kamanda et al. Reference Kamanda, Ndiweni, Imbayarwo-Chikosi, Muvengwi and Musakwa2008). Information from these and similar initiatives can help to identify locations within the mopane woodlands where pools can be excavated and/or sustained by pumps in the dry season. Additional water sources would benefit other wildlife populations also, by expanding animal distributions and increasing productivity, survival rates, and fitness (Rosenstock et al. Reference Rosenstock, Ballard and Devos1999). Such interventions, however, need to be carefully planned to ensure compatibility with long-term landscape management and community welfare objectives as well as the natural mosaic of spatio-temporal variability in surface water (Redfern et al. Reference Redfern, Grant, Gaylard and Getz2005; Smit2013). thank the Foundation, A. G. Leventis Foundation, and Club (ABC) for financial support, Birdfor the administrative support to manage the ABC grant, and the Department of and Wildlife for the research permits. We also thank three referees for their insightful comments on the submitted draft of this paper. Special appreciation goes to the 13 pool monitors in Ngwezi, Machile, Mulanga, and Parks, whose commitment and contribution to this work was invaluable. We dedicate this paper to water pool monitors and guardians of the Black-cheeked Lovebirds who passed away before the completion of this study. Supplementary material The supplementary material for this 10. 1017/S0959270924000261. 8. Barcelona: Lynx Edicions, 477. 309. Redlands: ESRI. 136. revised 2023. 1126. Range and study area of Black-cheeked Lovebird (BCL) Agapornis nigrigenis as documented by Bird(2024). View in content Table 1.

Output: contrast


Input: Claim: Cave paintings offer a glimpse into prehistoric human expression. [SEP] Text: Informed consent was obtained from all participants and ethical approval for the study was obtained from University of Committee (Medical). Measurement of cardiometabolic variables Fasting lipograms and glucose were measured on the Cobas® 8100 (Roche, Basel, Switzerland) auto-analyzer in the routine Laboratory of the (NHLS) based at Hospital. The coefficients of variation (CV) for cholesterol were 1. 6 % and 1. 4 %, for triglycerides were 2. 7 % and 1. 1 %, for HDL-C were 0. 9 % and 0. 9 % and for LDL-C were 0. 9 % and 0. 9 % for low and high control samples, respectively. The detection limit for cholesterol, triglycerides and LDL-C was 0. 1 mmol/L, while for HDL-C it was 0. 08 mmol/L. Cell culture Monocyte isolation Human monocytes were isolated using a modified method based on that described by de Almeida and colleagues (17). Buffy coat was retrieved from 45 ml of blood in ethylenediaminetetraacetic acid (EDTA) tubes (), overlaid on Histopaque®-1077 (density 1. 077 g/ml, []) and centrifuged at 2360 x g for 10 min. Following centrifugation, the peripheral blood mononuclear cells (PBMCs) were isolated and plated into a 6 well cell culture plate (Nest Biotechnology, Wuxi, China) at a concentration of 2 x 106 cells per well in RPMI growth medium (Lonza Bioscience, Basel, Switzerland) supplemented with 10% fetal bovine serum, 25m2mM L-glutamine, and 100U/ml Penicillin/Streptomycin. The cells were maintained under standard conditions (in an incubator at 37°C and 5% CO2 in humidified air). After 24 hours of culture the monocytes adhered to the culture plate and non-adherent lymphocytes were removed by rinsing the culture plate with 2 ml growth medium and the isolated monocytes were subsequently converted to macrophages.

Output: pass


Input: Claim: CRISPR technology allows for precise editing of genomic sequences. [SEP] Text: The 25 clones were then sequenced by Sequencing (NGS) to ensure correct gene editing was achieved (Fig. 2). /Cas9 editing is usually bi-allelic. Our results showed a total of 41 alleles were edited while 9 alleles had no editing. These 41 edited alleles resulted in 21 (84%) modified clones with 7 genotypes (Fig. 2). Of the 25 clones, 5 alleles (10%) were edited through the HDR pathway and 36 alleles (72%) were edited through the NHEJ pathway. The most frequent editing was an A deletion at codon 225 through NHEJ pathway which resulted in 11 (44%) clones with homozygous A deletion (Clone 7, Fig. 2). /Cas9 editing generated 5 HLA-B*38:68Q alleles that resulted in one HLA-B*38:68Q homozygous clone and one HLA-B*38:01:01:01/B*38:68Q heterozygous clone. The sequencing of the entire HLA-B gene from the 5′ UTR to 3′ UTR of the HLA-B*38:68Q homozygous clone and a HLA-B*38:01:01:01/B*38:68Q heterozygous clone showed that other than the correct 9-nt deletion on codon 230–232 at exon 4, there was no non-specific gene editing detected in the rest of the HLA-B*38:68Q alleles in both clones (Fig. 3). Figure 2Single cell clones derived byCas9-mediated gene editing. Nucleotides highlighted in red are the position with the 9-nt deletion. (PAM) sequence is highlighted in green. “. ” indicates the deletion. The frequency of each single cell derived clone as well as the relative percentage of wild type (WT), NHEJ, and HDR were listed (n = 25). Full size imageFigure 3Confirmation of HLA-B*38:68Q gene editing by NGS sequencing. (A) HLA-B*38:01:01:01 homozygous; (B) HLA-B*38:68Q/B*38:01:01:01 heterozygous; (C) HLA-B*38:68Q homozygous.

Output: corroborate


Input: Claim: Limnology has no relevance for human water usage or conservation efforts. [SEP] Text: Back to Role of in Water Usage: from Cape Town, Last registered on November 06, 2015 History Pre-of in Water Usage: from -0000921 Initial registration date November 06, 2015 Initial registration date is when the trial was registered. It corresponds to when the registration was submitted to the Registry to be reviewed for publication. First published November 06, 2015, 12:10 published corresponds to when the trial was first made public on the Registry after being reviewed. of Economics, University of (s) development Start date 2014-06-01 End date 2016-12-31 & code(s) C93; O13; Q25; Q56 has moved from a situation of water stress to one of water scarcity as a result of pressures from population growth, climate change, over-consumption and urbanization. We study the effectiveness of non-price instruments at promoting water conservation in the residential sector in Cape Town, South Africa's second largest city. Using inserts in monthly customer bills, we test nine behavioral messages (treatments) in a randomized control trial on 412,489 households. The treatments are classified into four groups: public-good framing (voluntary contributions to a common goal/good), financial framing (focusing on the financial losses/gains associated with inefficient/efficient water usage), social-comparison framing (where household consumption is compared to that of other households) and social-recognition framing (recognizing water conservation). External Link(s) Citation Brick, Kerri et al. 2015. "of in Water Usage: from Cape Town, South Africa. ". November 06. Brick, Kerri et al. 2015. "of in Water Usage: from Cape Town, South Africa. ". November 06. Sponsors & is information in this trial unavailable to the public. Use the button below to request access. (s) This study considering the role of behavioral interventions in promoting water conservation is to be rolled-out to residential consumers in the City of Cape Town, South Africa. Our unit of randomization is the household and our outcome variable is household water consumption (kilo-liters per month). Participating households have been allocated into nine treatment groups (which receive the interventions or behavioral messages) and a control group (which receives no intervention). The nine behavioral messages are to be sent via billing inserts with the monthly municipal water bills. The interventions can be described in terms of: A. - Increasing salience B.

Output: contrast


Input: Claim: Cave paintings offer a glimpse into prehistoric human expression. [SEP] Text: One well of a 6-well culture plate (containing a coverslip) was used for determining the monocyte purity by incubating the cells with a primary antibody that binds to calprotectin, an intracytoplasmic protein expressed by monocytes and macrophages (18). Random sections of the cover slip were chosen, and 100 cells were counted. Both stained and unstained cells were counted, and the percentage of stained cells was calculated. Macrophage differentiation After 24 hours of culture (see above), monocytes were differentiated into macrophages by incubation with 1ml growth media supplemented with macrophage colony-stimulating factor (M-CSF (100 ng/ml) []) for 5 days under standard conditions. Macrophage differentiation was confirmed through microscopic detection of morphological changes (elongated or stellate morphology) (19). Quantification of scavenger receptor expression by quantitative real time was extracted from macrophages using the Qiagen RKit (Qiagen, Hilden, Germany) and D(Sigma) was used to eliminate DNA from RNA template according to the manufacturer’s instructions. Complementary DNA (cDNA) was synthesized from RNA using the RevertStrand cDNA Synthesis kit (). Real-Time PCR was performed using Maxima SYBR® Green master mix (). The quantification of the levels of expression of the scavenger receptor genes was carried out on the StepOnePlus real time PCR system (). Relative gene expression of SRs CD36, CD68 and MSR1 was calculated using the ∆∆Ct relative quantification method with beta (β)-actin used as the normalization control. The cDNA amplification of CD36, CD68 and MSR1 was performed using the following conditions over 45 cycles: initial denaturation at 95°C for 2 minutes; denaturation at 95°C for 30 seconds; annealing at 55°C for 30 seconds; extension at 72°C for 1 minute. The primer sequences used for quantification were as follows: CD36 forward primer 5'--3', reverse primer 5'--3'; CD68 forward primer 5'- -3', reverse primer 5’--3'; MSRI forward primer 5'- -3', reverse primer 5'-- 3'; beta actin forward primer 5'--3', reverse primer 5'- -3'.

Output: pass


Input: Claim: CRISPR technology allows for precise editing of genomic sequences. [SEP] Text: Full size imageDiscussionIn this study, we reported efficient HLA-B*38:01:01:01 gene modification and expression in an EBV B cell line using theCas9 system. We successfully introduced gene editing in 84% of clones and achieved precise deletion at codon 230–232 at exon 4 in 5 alleles. Similar to other publications, theCas9 gene editing of HLA-B*38:01:01:01 involved DNA repair via either NHEJ or HDR pathway27. However, even in the presence of guided DNA templates, 72% of gene editing was through the NHEJ repair pathway compared to 10% in HDR pathway. HDR pathway provides desired repair of the target DNA in the presence of template DNA. The low incidence of HDR makes the selection of precise gene editing challenging. In order to achieve higher HDR gene editing efficiency, the DSBs induced byCas9 nuclease should be in close proximity to the edit site18. The homologous recombination rate could be increased with larger flanking sequences, therefore standard gene deletion/disruption protocols typically use flanking regions over 1 kb on either side of the target gene to increase HDR28. The evidence of using cell lines deficient in NHEJ pathways increased the levels of HDR suggesting these two pathways are competitive29. Recent studies have demonstrated the use of KU70, KU80 or DNA ligase IV to suppress key NHEJ molecule can increase HDR pathway30. Our study successfully demonstrated that the deletion at codon 230–232 at exon 4 in HLA-B*38:01:01:01 reduced HLA-B38 antigen expression on the cell surface usingCas9 system. The expression of the HLA-B*38:01:01:01 homozygous allele was 2. 2 fold higher than HLA-B*38:68Q/B*38:01:01:01 heterozygous cells. Further, HLA-B*38:68Q/B*38:01:01:01 protein expression in heterozygous cells was >2. 0 fold higher compared to HLA-B*38:68Q homozygous cells. Similar results were found using the anti-B38 polyclonal antibody.

Output: corroborate


Input: Claim: Limnology has no relevance for human water usage or conservation efforts. [SEP] Text: & Losses C. D. E. F. Date 2015-11-06 Date 2016-05-05 (end points) Monthly water consumption (in kiloliters) at the household level. (explanation) (end points) (explanation) The eligible population for this study is all residential households with access to piped water in the City of Cape Town. We are excluding commercial customers and only including those classified as “domestic” by the City. Furthermore, as we are limiting our sample to free standing houses, we are also excluding households with bulk meters. This obviates issues regarding flats where one meter number could be linked to multiple households. It further ensures that the recipient of the message is most likely one of the household members consuming water. We are excluding those customers consuming only in 1, as this is the lowest consumption band (6 kilolitres/month) and is supplied free by the City. These households are already consuming the lowest possible amount of water each month and thus we assume they are either the most vulnerable residents in the City of or are not residing in the household on a full-time basis (i. e. holiday home). If they are the most vulnerable residents in Cape Town, we want to safeguard against messaging them as it may be unethical (i. e. hazardous) to ask the lowest income residents to consume less water. However, as we are interested in how different behavioural messages affect consumption across all income groups, we are including households with “indigent” status in the treatments - provided they are consuming above 1. Indigent households in are those deemed most vulnerable and, according to the City’s policy, struggle to pay for public utilities. Given that those receiving an indigent grant receive varying levels of subsidization of utilities, or receive utilities free of charge, we chose not to include them in the financial incentives treatments (treatments 2, 3 and 4). We will not message them regarding financial savings as our code would significantly overestimate the financial savings associated with a reduction in water consumption. The sample size is the population of the City of excluding commercial and bulk accounts, apartment flats, and those consuming only within 1. This is a total expected sample size of 412,480 households: 113,312 are indigent and 299,168 are not indigent residents. We stratify the sample on suburbs (676) and tariff blocks (6 total).

Output: contrast


Input: Claim: Cave paintings offer a glimpse into prehistoric human expression. [SEP] Text: Statistical analysis Continuous variables with a normal or skewed distribution were expressed as means ± SD or median (interquartile range), respectively. Categorical variables were expressed as percentages. Continuous data were compared across the three ethnic groups using ANOVA, with skewed data log normalised before being analysed and paired comparisons performed using a Tukey post hoc test. Categorical variables were compared across the groups using a χ2 test. correlations of SR levels with gender, ethnicity, age, anthropometry, blood lipid and glucose levels were performed and variables with p < 0. 20 were included as independent variables in multivariable linear regression models. Multivariable linear regression analysis was performed using a backward stepwise model to find the main determinants of SR expression levels. A p < 0. 05 was considered significant. Statistical analyses were performed using Statistica software version 13. 0 (). Sample sizes were not calculated due to lack of data on ethnic differences in macrophage SR expression. The sample size chosen (10 subjects per group) was based on study infrastructural constraints, but with allowance for expansion of the sample size if significant differences were not observed with n = 10. ResultsMonocyte purity was assessed for all 30 isolates and was found to be 96. 0 ± 6. 0 %. The purity was compared across the 3 ethnic groups and no statistical differences were observed (p = 0. 28). Table 1 shows the data comparison across ethnicity. Africans had a significantly higher BMI than Europeans (p = 0. 03). Hip circumference was significantly higher in Africans as compared to Europeans (p = 0. 01) and Indians (p = 0. 004). No differences were observed in terms of fasting lipograms (total cholesterol, and triglycerides) and glucose between the ethnic groups. Table 1 Data comparison across ethnic groups (n = 10) Europeans (n = 10) Indians (n = 10) Males (%) 50. 0 50. 0 50. 0 Age (years) 46. 9 ± 5. 1 53. 8 ± 10. 6 54. 4 ± 9. 8 BMI (kg/m2) 30. 2 (27. 8, 40. 4) 24. 1 (20. 8, 25. 9) * 26. 9 (21. 1, 29. 0) Waist (cm) 96. 6 ± 9. 1 88. 7 ± 10. 6 85. 2 ± 15.

Output: pass


Input: Claim: CRISPR technology allows for precise editing of genomic sequences. [SEP] Text: Failure to identify HLA null alleles in donors may cause severe GVHD in HSCT7. Increasing knowledge of the expression level of HLA variant alleles will help to improve the understanding of HLA allogenicity in both HSCT and solid organ transplantation. /Cas9 system provides an effective tool to study the expression level of these variant HLA alleles. In addition,Cas9 can also introduce insertions and deletions at the UTRs, exons and introns to study the regulations and functions of HLA genes. /Cas9 has been used to facilitate correction of mutated genes in various diseases. /Cas9 has been used to treat single nucleotide polymorphism in the β-globulin gene to treat sickle cell diseases in a mouse model35. The chimeric antigen receptor (CAR) modified T cells have been applied to various cancers, especially in B cell hematologic malignancies36. With the applicationCas9 system, Liu et al. 37 have successfully down regulated the expression of HLA class I and TCRα to generate a universal chimeric antigen receptor (CAR) T cells. /Cas9 technology has also come under the spotlight in transplantation. Entry of human immunodeficiency virus (HIV) into target cells requires both CD4 and CCR5 receptors38. A 32-base pair deletion in CCR5 (CCR5-Δ32) is associated with reduced HIV transmission risk and delayed disease progression39. In HIV+ patients with hematological malignancies, gene editing usingCas940 has been used to generate homozygous CCR5-Δ32 deletion in CD34+ cells to introduce HIV resistance. Currently, there are several ongoing clinical trials evaluating the safety of transplantation of modified CCR5-Δ32 CD34+ cells in HIV+ patients with hematological malignances41,42. In conclusion, theCas9 system is a powerful gene editing tool that can be used to study HLA gene expression and function and improve HLA matching in hematopoietic stem cell and solid organ transplantation. Modification of HLA gene expression byCas9 also promises to provide new approaches for cellular therapies in transplantation.

Output: corroborate


Input: Claim: Limnology has no relevance for human water usage or conservation efforts. [SEP] Text: Back to Role of in Water Usage: from Cape Town, Last registered on November 06, 2015 History Pre-of in Water Usage: from -0000921 Initial registration date November 06, 2015 Initial registration date is when the trial was registered. It corresponds to when the registration was submitted to the Registry to be reviewed for publication. First published November 06, 2015, 12:10 published corresponds to when the trial was first made public on the Registry after being reviewed. Last updated November 06, 2015, 9:25 updated is the most recent time when changes to the trial's registration were published. of Economics, University of (s) development Start date 2014-06-01 End date 2016-12-31 & code(s) C93; O13; Q25; Q56 has moved from a situation of water stress to one of water scarcity as a result of pressures from population growth, climate change, over-consumption and urbanization. We study the effectiveness of non-price instruments at promoting water conservation in the residential sector in Cape Town, South Africa's second largest city. Using inserts in monthly customer bills, we test nine behavioral messages (treatments) in a randomized control trial on 412,489 households. The treatments are classified into four groups: public-good framing (voluntary contributions to a common goal/good), financial framing (focusing on the financial losses/gains associated with inefficient/efficient water usage), social-comparison framing (where household consumption is compared to that of other households) and social-recognition framing (recognizing water conservation). External Link(s) Citation Brick, Kerri et al. 2015. "of in Water Usage: from Cape Town, South Africa. ". November 06. Brick, Kerri et al. 2015. "of in Water Usage: from Cape Town, South Africa. ". November 06. Sponsors & is information in this trial unavailable to the public. Use the button below to request access. (s) This study considering the role of behavioral interventions in promoting water conservation is to be rolled-out to residential consumers in the City of Cape Town, South Africa. Our unit of randomization is the household and our outcome variable is household water consumption (kilo-liters per month). Participating households have been allocated into nine treatment groups (which receive the interventions or behavioral messages) and a control group (which receives no intervention).

Output: contrast


Input: Claim: Cave paintings offer a glimpse into prehistoric human expression. [SEP] Text: 7 Hip (cm) 115. 5 ± 14. 9 100. 6 ± 6. 6* 98. 7 ± 9. 0** Cholesterol (mmol/L) 4. 6 ± 0. 7 5. 1 ± 0. 6 5. 2 ± 0. 8 HDL-C (mmol/L) 1. 4 (1. 0, 1. 7) 1. 5 (1. 3, 1. 9) 1. 5 (1. 1, 1. 6) LDL-C (mmol/L) 3. 0 ± 0. 6 3. 5 ± 0. 5 3. 5 ± 0. 9 Triglycerides (mmol/L) 0. 9 (0. 7, 1. 1) 0. 7 (0. 6, 1. 2) 0. 9 (0. 7, 1. 1) Glucose (mmol/L) 4. 7 ± 0. 8 5. 1 ± 0. 6 5. 3 ± 1. 0 Data given as mean ± SD or median (interquartile range); *p < 0. 05, **p < 0. 005 vs African. Scavenger receptor gene expression from macrophages isolated from African, European and Indian populations Scavenger receptor gene expression were compared across ethnic groups (Fig. 1). Expression of CD36 was significantly lower in African (p = 0. 0005) and Indian (p = 0. 04) compared to European participants. The expression levels of CD68 in Indian (p = 0. 007) participants were significantly lower compared to European participants whilst MSR1 expression was higher in European (p = 0. 0002) and Indian (p = 0. 0001) than African participants. Determinants of macrophage SR gene expression Note that Table 2 and 3 the n = 28 instead of n = 30, this is because when we performed normal distribution of scavenger receptors, we found outliers and we removed them form our data. Table 2 outlines the factors that are associated with the scavenger receptor expression in macrophages. Univariate correlation analysis of CD36 gene expression demonstrated positive associations with age (p = 0. 01), CD68 expression (p < 0. 001) and MSR1 expression (p = 0. 02). whilst CD68 gene expression associated positively with serum triglycerides (p = 0. 04).

Output: pass


Input: Claim: CRISPR technology allows for precise editing of genomic sequences. [SEP] Text: Material and MethodsHLA-B*38:68Q was submitted to the GenBank database (accession number MF069211) and IMGT/HLA database (submission number HWS10028807) with full genomic allele sequence as a questionable allele due to its unknown surface expression by our center in 201743. Research approval for performingCAS9 on the sample was granted by the (IRB#14–000516). Cell linesAn HLA-B*38:01:01:01 homozygous EBV transformed B cell line TEM665 (from 9057, Fig. 1B) was selected for gene editing, and in addition, a Bw6 homozygous EBV B cell line AOH749 (from 9004) and/or K562 (from ) which lacks HLA expression were used as negative controls for monoclonal or polyclonal antibody test. AOH749 expresses HLA A31, B65, Bw6, C8, DR1, DQ5, DP3 and DP0401, which does not cross reactive to the anti-Bw4 antibody. All cell lines were cultured in RPMI-1640 () containing 20% FBS (Omega, USA), 1% (Corning, USA), 1% Solution (Corning, USA), 1% Gibco™ (100 mM) (ThermoFisher, USA), 1% Gibco™ MEM Non-(ThermoFisher, USA), and 0. 1% Gibco™ 2-Mercaptoethanol (55 mM) (ThermoFisher, USA). All cultures were maintained in 5% CO2, 37 °C. crRNA design and Cas9 Ribonucleoproteins (RNPs) complex formationOligonucleotides of crRNA sequences were designed with the and Design Tool. crRNAs (Synthesized by ThermoFisher, USA) and tracrRNA from TrueGuide™ Synthetic gRNA kit (ThermoFisher, USA) were re-suspended using DNA pH 8. 0 (Teknova, USA) to make a concentration of 100 pM.

Output: corroborate


Input: Claim: Limnology has no relevance for human water usage or conservation efforts. [SEP] Text: The nine behavioral messages are to be sent via billing inserts with the monthly municipal water bills. The interventions can be described in terms of: A. - Increasing salience B. & Losses C. D. E. F. Date 2015-11-06 Date 2016-05-05 (end points) Monthly water consumption (in kiloliters) at the household level. (explanation) (end points) (explanation) The eligible population for this study is all residential households with access to piped water in the City of Cape Town. We are excluding commercial customers and only including those classified as “domestic” by the City. Furthermore, as we are limiting our sample to free standing houses, we are also excluding households with bulk meters. This obviates issues regarding flats where one meter number could be linked to multiple households. It further ensures that the recipient of the message is most likely one of the household members consuming water. We are excluding those customers consuming only in 1, as this is the lowest consumption band (6 kilolitres/month) and is supplied free by the City. These households are already consuming the lowest possible amount of water each month and thus we assume they are either the most vulnerable residents in the City of or are not residing in the household on a full-time basis (i. e. holiday home). If they are the most vulnerable residents in Cape Town, we want to safeguard against messaging them as it may be unethical (i. e. hazardous) to ask the lowest income residents to consume less water. However, as we are interested in how different behavioural messages affect consumption across all income groups, we are including households with “indigent” status in the treatments - provided they are consuming above 1. Indigent households in are those deemed most vulnerable and, according to the City’s policy, struggle to pay for public utilities. Given that those receiving an indigent grant receive varying levels of subsidization of utilities, or receive utilities free of charge, we chose not to include them in the financial incentives treatments (treatments 2, 3 and 4). We will not message them regarding financial savings as our code would significantly overestimate the financial savings associated with a reduction in water consumption. The sample size is the population of the City of excluding commercial and bulk accounts, apartment flats, and those consuming only within 1.

Output: contrast


Input: Claim: Cave paintings offer a glimpse into prehistoric human expression. [SEP] Text: Gene expression of MSR1 was associated positively with serum LDL-C (p = 0. 001) and plasma glucose (p = 0. 005). Table 2 Univariate correlations for macrophage scavenger receptor expression value (p-value) CD36 (n = 28) CD68 (n = 28) MSR1 (n = 28) Age (Years) 0. 455 (0. 013) 0. 130 (0. 500) 0. 256 (0. 189) BMI (kg/m2) -0. 260 (0. 173) -0. 084 (0. 666) -0. 266 (0. 172) Waist (cm) 0. 027 (0. 891) 0. 119 (0. 537) -0. 026 (0. 896) Hip (cm) -0. 239 (0. 212) 0. 030 (0. 878) -0. 334 (0. 082) Cholesterol (mmol/L) 0. 291(0. 226) -0. 039 (0. 766) 0. 345 (0. 072) HDL-C (mmol/L) -0. 010 (0. 895) -0. 117 (0. 517) -0. 273 (0. 159) LDL-C (mmol/L) 0. 333 (0. 114) -0. 023 (0. 915) 0. 575 (0. 001) Triglycerides (mmol/L) 0. 112 (0. 721) 0. 377 (0. 031) -0. 022 (0. 910) Glucose (mmol/L) 0. 105 (0. 498) -0. 117 (0. 680) 0. 516 (0. 005) CD36 expression 0. 607 (0. 000) 0. 424 (0. 015) CD68 expression 0. 607 (0. 000) 0. 072 (0. 717) MSR1 expression 0. 424 (0. 015) 0. 072 (0. 717) Table 3 Multivariate regression models for macrophage scavenger receptor expression Dependent variable Independent variables with β-coefficient (p-value) Adjusted R2 (p-value) CD36 (n=28) CD68 (n=28) MSR1 (n=28) Age; 0.

Output: pass


Input: Claim: CRISPR technology allows for precise editing of genomic sequences. [SEP] Text: Skip to Content eLife home page your research Search by keyword or author Reset form my search to Chromosomes and Research 10. 7554/eLife. 41769 Open access Copyright information Version of RecordMay 24, 2019 Open annotations (there are currently 0 annotations on this page). 17,166 views 1,675 downloads 239 1769. Copy to clipboard Download BibTeX Download. RIS Full text Figures and data Peer review Side by side files 12 figures, 5 videos, 1 table and 8 additional files 1 with 3 supplements Download asset Open asset The Sox2 a Model for Visualization of Enhancer-in Stem Cells. (A) To visualize chromosome loci in living cells, we have used tetO/TetR and cuO/CymR genetic labels. Our pipeline for insertion of these labels into the mouse genome is shown. -Cas9 is used to place an attP integrase landing site. Second, a targeting plasmid bearing the compatible attB sequence, the tetO or cuO array, and a selection cassette is introduced along the integrase (Int) to mediate site-specific integration. The selection cassette can then be subsequently removed by Cre/Flp recombinase. (B) The Sox2 locus in mouse ESCs. Genomic browser tracks of epigenomic and expression data demonstrate high levels of histone acetylation, RNA polymerase II, and transcription factor (OCT4, SOX2, ) and cohesin (RAD21) occupancy at Sox2 and the distal Sox2 enhancer (tan boxes). Data from 4C and HiC experiments demonstrate chromosomal contacts at the Sox2 locus. For 4C data, read density indicates contact frequency with a fixed position near the Sox2 promoter (red triangle). Y-axis for browser tracks is reads per million. For HiC, all pairwise contact frequencies are shown using a heatmap. The intensity of each pixel represents the normalized number of contacts detected between a pair of loci. The maximum intensity is indicated in red square. At bottom, locations of the cuO- and tetO-arrays for the three cell lines utilized for this study. Sox2-8CcuO/+; Sox2-117TtetO/+ (Sox2-SCR) ESCs were used to track Sox2/SCR location.

Output: pass


Input: Claim: Bees always live in colonies, with no exceptions. [SEP] Text: social insect colonies exhibit a remarkable ability for recognizing group members via colony-specific cuticular pheromonal signatures. Previous work suggested that in some ant species, colony-specific pheromonal profiles are generated through a mechanism involving the transfer and homogenization of cuticular hydrocarbons (CHCs) across members of the colony. However, how colony-specific chemical profiles are generated in other social insect clades remains mostly unknown. Here we show that in the honey bee (Apis mellifera), the colony-specific CHC profile completes its maturation in foragers via a sequence of stereotypic age-dependent quantitative and qualitative chemical transitions, which are driven by environmentally-sensitive intrinsic biosynthetic pathways. Therefore, the CHC profiles of individual honey bees are not likely produced through homogenization and transfer mechanisms, but instead mature in association with age-dependent division of labor. Furthermore, non-nestmate rejection behaviors seem to be contextually restricted to behavioral interactions between entering foragers and guards at the hive entrance. eLife digest Honey bees are social insects that live in large groups called colonies, within structures known as hives. The young adult bees stay within the hive to build nests and care for the young, while the older bees leave the hive to forage for food. Honey bees store food and other valuable resources in their hives, so they are often targeted by predators, parasites and ‘robber’ bees from other colonies. Therefore, it is important for bees to determine whether individuals trying to enter the nest are group members or intruders. While it is known that social insects use blends of waxy chemicals called cuticular hydrocarbons to identify group members at the entrance to the colony, it is not clear how members of the same colony acquire a similar blend of cuticular hydrocarbons. Some previous work suggested that in some ant species (which are also social insects), colony members exchange cuticular hydrocarbons with each other so that all members of the colony are covered with a similar blend of chemicals. However, it was not known whether honey bees also share cuticular hydrocarbons between colony members in order to identify members of a hive. Vernier et al. used chemical, molecular and behavioral approaches to study the cuticular hydrocarbons found on honey bees. The results show that, rather than exchanging chemicals with other members of their colony, individual bees make their own blends of cuticular hydrocarbons.

Output: contrast


Input: Claim: Cave paintings offer a glimpse into prehistoric human expression. [SEP] Text: 33 (0. 02) LDL-C; 0. 29 (0. 03) CD68; 0. 57 (<0. 001) Europeans; 0. 56 (<0. 001) Triglycerides; 0. 45 (0. 005) Africans; -0. 73 (<0. 001) HDL-C; -0. 33 (0. 003) LDL-C; 0. 27 (0. 015) 0. 55 (<0. 001) 0. 41 (<0. 001) 0. 77 (<0. 001) Using the data from Fig. 1 and Table 2, backward, stepwise linear multivariate regression analysis of gene expression was performed, and the results are shown in Table 3. The CD36 scavenger receptor expression correlated with age (p = 0. 02), LDL (p = 0. 03) and CD68 expression (p < 0. 001) whilst CD68 expression correlated positively with triglycerides (p = 0. 005) and European ethnicity (p < 0. 001). Gene expression of MSR1 correlated positively with LDL (p = 0. 015) and negatively with HDL (p = 0. 003) and African ethnicity (p < 0. 001). DiscussionThe current study shows clear associations of serum lipid levels and ethnicity with macrophage SR gene expression. Thus, CD36 and MSR1 expression were both positively associated with serum LDL levels, and MSR1 was also negatively associated with HDL. Gene expression of CD68 was positively associated with serum triglyceride levels. In terms of the association of ethnicity with SR expression, CD68 was expressed at higher levels in subjects with European ancestry, whilst MSR1 was expressed at lower levels in African subjects. When comparing data between the 3 ethnic groups, the African participants had significantly higher levels of both BMI and hip circumference compared to the other groups. It is known that obesity is more prevalent in African than other ethnic groups in South Africa, particularly in females (20). In addition, LDL and total cholesterol, although not statistically significantly so, were lower in the African compared to the other groups. Similar ethnic differences in lipid profiles have been reported in other studies (21, 22). Scavenger receptors MSR1and CD36 have been shown to be the primary receptors responsible for the uptake of modified LDL into macrophages leading to foam cell formation (23) (2).

Output: pass


Input: Claim: CRISPR technology allows for precise editing of genomic sequences. [SEP] Text: Cas9 gene editing has revolutionised loss-of-function experiments in Leishmania, the causative agent of leishmaniasis. lack a functional non-homologous DNA end joining pathway however, obtaining null mutants typically requires additional donor DNA, selection of drug resistance-associated edits or time-consuming isolation of clones. Genome-wide loss-of-function screens across different conditions and across multiple Leishmania species are therefore unfeasible at present. Here, we report aCas9 cytosine base editor (CBE) toolbox that overcomes these limitations. We employed CBEs in Leishmania to introduce STOP codons by converting cytosine into thymine and created for CBE primer design in kinetoplastids. Through reporter assays and by targeting single- and multi-copy genes in L. mexicana, L. major, L. donovani, and L. infantum, we demonstrate how this tool can efficiently generate functional null mutants by expressing just one single-guide RNA, reaching up to 100% editing rate in non-clonal populations. We then generated a Leishmania-optimised CBE and successfully targeted an essential gene in a plasmid library delivered loss-of-function screen in L. mexicana. Since our method does not require DNA double-strand breaks, homologous recombination, donor DNA, or isolation of clones, we believe that this enables for the first time functional genetic screens in Leishmania via delivery of plasmid libraries. Editor's evaluation Leishmanias cause a wide spectrum of diseases in many animals: in humans, symptoms range from disfiguring skin lesions to lethal visceral infections. Knowledge of gene function has been limited by aneuploidy, as well as the absence of RNAi (in most but not all species) and of non-homologous DNA end joining (which limits the utility of Crispr-Cas systems). In this paper, the authors describe the successful testing and application of a new method for functional screens in Leishmania, in which targeted cytosine base editing is used to introduce premature stop codons within open reading frames. The advantages and disadvantages of this approach, in comparison with others, are described in a balanced and careful way. This is a very important new tool for researchers in Leishmania, and might also serve as a model for other poorly tractable species.

Output: corroborate


Input: Claim: Bees always live in colonies, with no exceptions. [SEP] Text: As a bee ages it makes different blends of cuticular hydrocarbons, and by the time it starts to leave the hive to forage it makes a blend that is specific to the colony it belongs to. The production of this final blend is influenced by the environment within the hive. Thus, the findings of Vernier et al. indicate that honey bees guarding the entrance to a hive can only identify non-colony-member forager bees as intruders, rather than any non-colony-member bee that happens upon the hive entrance. Honey bees play an essential role in pollinating many crop plants so understanding how these insects maintain their social groups may help to improve agriculture in the future. Furthermore, this work may aid our understanding of how other social insects interact in a variety of biological situations. ability to recognize ‘self’ plays an important role in regulating diverse processes across biological organizational levels (Tsutsui, 2004). Analogous to the acquired immunity system, which depends on self-recognition at the cellular and molecular levels (Boehm, 2006), adaptive organismal social interactions often depend on the recognition of kin and/or group-members to increase cooperation or to suppress inbreeding (Hamilton, 1964a; Hamilton, 1964b; Pusey and Wolf, 1996; Trivers, 1971; West et al. , 2007; Wilkinson, 1988). One remarkable example of organismal recognition of ‘self’ comes from colonies of social insects, which depend on a robust non-nestmate discrimination system (more commonly called ‘nestmate recognition’) to prevent the loss of expensive resources to non-nestmates, and to maintain overall colony integrity (Hefetz, 2007; van Zweden and D’Ettorre, 2010). As in other self-recognition systems, theoretical models suggest that nestmate recognition in social insect colonies depends on the ability of individual colony members to reliably match colony-specific phenotypic cues, or ‘labels’, carried by other colony members, to stored neural ‘templates’ (Buckle and Greenberg, 1981; Errard, 1994; Gamboa et al.

Output: contrast


Input: Claim: Cave paintings offer a glimpse into prehistoric human expression. [SEP] Text: The interaction of CD68 with modified lipoproteins is still controversial and their relevance is still unclear (2). However, a study by Tsukamoto and colleagues (2002) reported that CD68 may play a crucial role in foam cell formation through uptake of oxLDL by macrophages (24). In our study CD36 expression was lower in African and Indian compared to European participants, whereas the expression of CD68 was lower in Indian compared to European participants. The expression of the MSR1 gene was higher in European and Indian than African participants. Therefore, lower levels of expression of CD36 and MSR1 in African participants mirror a lower level of atherosclerotic disease observed in this population (15). Scavenger receptor expression levels correlated with baseline fasting lipid profiles, ethnicity and age. Thus, CD36 expression levels correlated positively with LDL-C, CD68 expression and age, whilst CD68 expression correlated positively with triglycerides and European ethnicity. The expression of the MSR1 gene correlated positively with LDL-C, and negatively with both HDL-C and African ethnicity. The positive association of serum LDL levels with macrophage SR gene expression is a novel finding and is supported by a study showing that LDL upregulates the expression of macrophage SRs in a murine macrophage cell line (25). Lipoproteins, particularly minimally modified LDL and oxidized LDL can modulate signal transduction and gene expression in macrophages and other cells. Thus, LDL stimulates phosphoinositide turnover and elevates cytosolic calcium levels in vascular smooth muscle cells. Minimally modified LDL induces the expression of monocyte adhesion molecules, tissue factor, macrophage chemotactic protein-1 (MCP-1) and M-CSF in endothelial cells (25). Furthermore, high levels of plasma LDL has the ability to activate signal transduction and gene expression in macrophages (26), through stimulation of phosphoinositide turnover. The association of serum lipid levels with macrophage SR expression that is described in the present study mirrors the effect of these lipid molecules on the risk of atherosclerosis. These data suggest that serum lipids may modulate atherosclerotic plaque formation by modifying the expression levels of scavenger receptors in circulating macrophages even before exposure to oxLDL.

Output: pass


Input: Claim: CRISPR technology allows for precise editing of genomic sequences. [SEP] Text: By simplifying previous labour-intensive gene replacement strategies (Cruz and Beverley, 1990),Cas9 has greatly improved loss-of-function experiments in Leishmania (Beneke et al. , 2017; Sollelis et al. , 2015; Zhang et al. , 2017; Zhang and Matlashewski, 2015). Over the past 8 years, multiple toolkits for kinetoplastids, including Leishmania, have emerged (Bryant et al. , 2019; Yagoubat et al. , 2020a; Pal and Dam, 2022), enabling specialised gene editing approaches, such as marker free (et al. , 2017) or inducible gene deletions (Damasceno et al. , 2020; Rico et al. , 2018; Yagoubat et al. , 2020b). In particular, the introduction of the PCR-based LeishGEdit toolbox (Beneke et al. , 2017; Beneke and Gluenz, 2019; Beneke and Gluenz, 2020) has boosted the number of successful loss-of-function studies in Leishmania species (Bryant et al. , 2019; Yagoubat et al. , 2020a; Jones et al. , 2018). LeishGEdit enabled bar-seq screens in-which hundreds of barcoded mutants are simultaneously phenotyped by pooling them together (Beneke and Gluenz, 2020; Baker et al. , 2021; Beneke et al. , 2019). This approach allows for precise deletion of single-copy or tandem arrayed multi-copy genes and since it produces complete knockouts, the number of false positives and negatives in any subsequent screening format is limited. Furthermore, the approach is scalable and it is therefore not surprising that this has led to a genome-wide loss-of-function screening proposal. Using the LeishGEdit method, the LeishGEM project ( attempts to knockout nearly all genes in the Leishmania mexicana genome and plans to uncover key virulence factors by screening barcoded mutants across 170 pooled populations. While these knockout pools will surely be a major resource for the research community in the future, their applications are limited. Cell pools are logistically difficult to share and it is unclear how well frozen pools recover and if they can be expanded after freezing without losing their uniformity.

Output: corroborate


Input: Claim: Bees always live in colonies, with no exceptions. [SEP] Text: , 1986; Getz, 1982; Hölldobler and Michener, 1980; Lacy and Sherman, 1983; Reeve, 1989; Tsutsui, 2004; van Zweden and D’Ettorre, 2010). In some social insect species, the cues used in recognizing individual members of the colony have been reported to be visual (Baracchi et al. , 2015), but in most cases are thought to be chemical (van Zweden and D’Ettorre, 2010). Cuticular hydrocarbons (CHCs), which evolved to function as hydrophobic, anti-desiccant barriers in terrestrial arthropods, have been co-opted to also function as pheromones in diverse insect communication systems, including nestmate recognition in social insect species (Chung and Carroll, 2015; van Zweden and D’Ettorre, 2010). Whether the overall profile, or more specific components of it, represent the actual nestmate recognition cue remains unknown. However, previous studies have indicated that variations in the relative amounts of each compound in the CHC profile across individuals from different colonies are likely sufficient for the chemical recognition of nest membership (van Zweden and D’Ettorre, 2010). Nevertheless, how large groups of hundreds to thousands of individuals coordinate the production and recognition of a robust colony-specific chemical cue remains unknown for most species. Because members of social insect colonies are often genetically related, it was initially assumed that the production of similar colony-specific pheromones by individual colony members is intrinsically driven by shared allelic variants (Crozier and Dix, 1979; Getz, 1982; Getz, 1981). However, empirical studies revealed that, surprisingly, in many social insect species colony and social environmental factors play the most dominant role in defining colony-specific cues, and can often mask genetic relatedness (Breed et al. , 1988; Downs and Ratnieks, 1999; Heinze et al. , 1996; Lahav et al. , 2001; Liang and Silverman, 2000; Singer and Espelie, 1996; Stuart, 1988). Although these colony ‘environmental’ factors remain unknown for most social insect species, it has been suggested that contributions from nest building materials (Breed et al. , 1988; Couvillon et al. , 2007; D'ettorre et al.

Output: contrast


Input: Claim: Cave paintings offer a glimpse into prehistoric human expression. [SEP] Text: Our findings were limited by the relatively small sample size however, we were still able to observe differences in SR levels across ethnicities and significant associations within the regression models. The other limitation is the analysis of associations between in vitro data i. e. SR expression, and in vivo data i. e. serum lipid levels. In performing these analyses we are assuming that lipid levels have an effect on macrophage SR expression that is not modified by monocyte isolation and cell culture conditions. Despite this limitation, it is interesting to note that the associations observed between serum lipid levels and in vitro macrophage SR expression are supported by other studies from the literature and are scientifically plausible. ConclusionOur data suggests that serum lipids may modulate atherosclerotic plaque formation by modifying the expression levels of scavenger receptors in macrophages even before exposure to oxLDL in the intima. Furthermore, ethnic differences in atherosclerotic plaque formation may be mediated through differential CD68 and MSR1 SRs expression levels within macrophages. AbbreviationsFC: foam cells; OxLDL: oxidized low-density lipoprotein; SRs: scavenger receptors; MSR1: macrophage scavenger receptor 1; M-CSF: macrophage-colony stimulating factor; LDL-C: low density lipoprotein- cholesterol; HDL-C: high density lipoprotein- cholesterol; BMI: body mass index; NHLS: Laboratory Service; CV: coefficients of variation; EDTA: ethylenediaminetetraacetic acid; PBMCs: peripheral blood mononuclear cells; MCP-1: macrophage chemotactic protein-1Declarations41. Scavenger receptors class A-I/II and CD36 are the principal receptors responsible for the uptake of modified low density lipoprotein leading to lipid loading in macrophages. J Biol Chem. 2002;277(51):49982–8. . Reversible accumulation of cholesteryl esters in macrophages incubated with acetylated lipoproteins. J Cell Biol. 1979;82(3):597–613. . CD36: a class B scavenger receptor involved in angiogenesis, atherosclerosis, inflammation, and lipid metabolism. J Clin Investig. 2001;108(6):785–91. .

Output: pass


Input: Claim: CRISPR technology allows for precise editing of genomic sequences. [SEP] Text: Our aim was to overcome these limitations by bypassing the DSB repair machinery and instead directly editing suitable codons into STOP codons by using aCas9 cytosine base editor (CBE) (Figure 1A). This strategy has previously been used for large-scale and genome-wide loss-of-function screens (Kuscu et al. , 2017; Després et al. , 2020). CBEs are comprised of a cytidine deaminase domain fused to an impaired form of Cas9. The Cas9 module provides precise targeting of the CBE to a site of interest, where the cytidine deaminase can directly convert cytosine to thymine without requiring DSBs, HR, or donor DNA templates (Huang et al. , 2021). Specifically, the deaminase domain catalyses a deamination reaction, converting cytidine (C) to uridine (U). Uridine is then repaired to thymidine (T) during DNA replication. To prevent base excision repair, which would convert uridine back to cytidine, Cas9 nickases (nCas9), such as D10A (Cong et al. , 2013), are additionally tethered to two monomers of a uracil glycosylase inhibitor (UGI) and further modifications of nuclear localisation signals (NLS), codon usage and the deaminase component have yielded highly efficient editors, such as the BE4max and AncBE4max (Koblan et al. , 2018; Komor et al. , 2016; Komor et al. , 2017). In addition, CBE editing rates can be further improved through insertion of a non-sequence-specific single-stranded (ss) DNA-binding domain (DBD) from the Rad51 protein between the Cas9 nickase and the deaminase domain (namely hyBE4max) (Figure 1A; Zhang et al. , 2020). Figure 1 Download asset Open asset Cytosine base editing (CBE) in Leishmania and other kinetoplastids.

Output: corroborate


Input: Claim: Bees always live in colonies, with no exceptions. [SEP] Text: , 2006; Espelie et al. , 1990; Singer and Espelie, 1996), the queen (Carlin and Hölldobler, 1988; Carlin and Holldobler, 1987; Carlin and Holldobler, 1986; Carlin and Hölldobler, 1983), and diet (Buczkowski et al. , 2005; Buczkowski and Silverman, 2006; Liang and Silverman, 2000; Richard et al. , 2004; Richard et al. , 2007) could, at least in part, provide unique chemical components to the chemical signature shared by colony members. Consequently, empirical and theoretical studies suggested that individual colony members acquire their colony-specific chemical signature largely through a homogenization process involving the exchange of relevant chemicals, including CHCs, through interactions between colony members or contact with nest building materials, often referred to as the ‘Gestalt’ model (Crozier and Dix, 1979). Empirical evidence in support of this model has been reported for a few ant species, which are known to transfer mixed blends of CHCs between individuals through trophallaxis and grooming via the action of the postpharyngeal gland (PPG) (Boulay et al. , 2000; Lenoir et al. , 2001; Meskali et al. , 1995; Soroker et al. , 1994; Soroker et al. , 1995b; van Zweden et al. , 2010). However, other studies suggest that such CHC homogenization processes might not fully represent how colony-specific chemical cues develop in all social insect species. For example, some ant species do not display robust trophallaxis behaviors, the main mode of chemical transfer across colony members (Soroker et al. , 1994; Soroker et al. , 1995b), and in others, the CHC profiles of individual colony members are likely modulated by genetic relatedness (Teseo et al. , 2014), age (Cuvillier-Hot et al. , 2001; Teseo et al. , 2014), and/or task (Martin and Drijfhout, 2009; Sturgis and Gordon, 2013; Wagner et al. , 2001; Wagner et al. , 1998).

Output: contrast


Input: Claim: Cave paintings offer a glimpse into prehistoric human expression. [SEP] Text: Mechanisms of cell signaling by the scavenger receptor CD36: implications in atherosclerosis and thrombosis. Assoc. 2010;121:206. Plasma levels of soluble CD36, platelet activation, inflammation, and oxidative stress are increased in type 2 diabetic patients. Biol Med. 2012;52(8):1318–24. Dai X-Y, Cai Y, Mao D-D, Qi Y-F, Tang C, Xu Q, et al. Increased stability of phosphatase and tensin homolog by intermedin leading to scavenger receptor A inhibition of macrophages reduces atherosclerosis in apolipoprotein E-deficient mice. J Cardiol. 2012;53(4):509–20. Mietus-. Regulation of scavenger receptor expression in smooth muscle cells by protein kinase C: a role for oxidative stress. Arteriosclerosis, thrombosis, and vascular biology. 1997;17(5):969–78. . Deletion of the murine scavenger receptor CD68. Journal of lipid research. 2011;52(8):1542–50. . The macrosialin promoter directs high levels of transcriptional activity in macrophages dependent on combinatorial interactions between PU. 1 and c-Jun. J Biol Chem. 1998;273(9):5389–99. . Minimally oxidized low-density lipoprotein increases expression of scavenger receptor A, CD36, and macrosialin in resident mouse peritoneal macrophages. Arteriosclerosis, thrombosis, and vascular biology. 1998;18(5):794–802. Shimada K. Immune system and atherosclerotic disease: heterogeneity of leukocyte subsets participating in the pathogenesis of atherosclerosis. Circulation journal: official journal of the Society. 2009;73(6):994–1001. . Cytokines, macrophage lipid metabolism and foam cells: implications for cardiovascular disease therapy. Progress in lipid research. 2011;50(4):331–47. Rafieian-. Atherosclerosis: process, indicators, risk factors and new hopes. International journal of preventive medicine. 2014;5(8):927. . Epidemiology of Atherosclerosis and the Potential to Reduce the of Atherothrombotic Disease. Circulation research.

Output: pass