text:
Isolation of potent SARS-CoV-2 neutralizing antibodies and protection from disease in a small animal model
View ORCID ProfileThomas F. Rogers1,2,*,
View ORCID ProfileFangzhu Zhao1,3,4,*,
View ORCID ProfileDeli Huang1,*,
View ORCID ProfileNathan Beutler1,*,
Alison Burns1,3,4,
View ORCID ProfileWan-ting He1,3,4,
View ORCID ProfileOliver Limbo3,5,
View ORCID ProfileChloe Smith1,3,
View ORCID ProfileGe Song1,3,4,
View ORCID ProfileJordan Woehl3,5,
View ORCID ProfileLinlin Yang1,
View ORCID ProfileRobert K. Abbott4,6,
Sean Callaghan1,3,4,
Elijah Garcia1,
View ORCID ProfileJonathan Hurtado1,4,7,
Mara Parren1,
Linghang Peng1,
View ORCID ProfileSydney Ramirez6,
James Ricketts1,
View ORCID ProfileMichael J. Ricciardi8,
Stephen A. Rawlings2,
View ORCID ProfileNicholas C. Wu9,
View ORCID ProfileMeng Yuan9,
View ORCID ProfileDavey M. Smith2,
View ORCID ProfileDavid Nemazee1,
View ORCID ProfileJohn R. Teijaro1,
View ORCID ProfileJames E. Voss1,
View ORCID ProfileIan A. Wilson3,4,9,
Raiees Andrabi1,3,4,
View ORCID ProfileBryan Briney1,4,7,
View ORCID ProfileElise Landais1,3,4,5,
View ORCID ProfileDevin Sok1,3,4,5,†,
View ORCID ProfileJoseph G. Jardine3,5,†,
View ORCID ProfileDennis R. Burton1,3,4,10,†
1Department of Immunology and Microbiology, The Scripps Research Institute, La Jolla, CA 92037, USA.
2Division of Infectious Diseases, Department of Medicine, University of California, San Diego, La Jolla, CA 92037, USA.
3IAVI Neutralizing Antibody Center, The Scripps Research Institute, La Jolla, CA 92037, USA.
4Consortium for HIV/AIDS Vaccine Development (CHAVD), The Scripps Research Institute, La Jolla, CA 92037, USA.
5IAVI, New York, NY 10004, USA.
6Center for Infectious Disease and Vaccine Research, La Jolla Institute for Immunology (LJI), La Jolla, CA 92037, USA.
7Center for Viral Systems Biology, The Scripps Research Institute, La Jolla, CA 92037, USA.
8Department of Pathology, George Washington University, Washington, DC 20052, USA.
9Department of Integrative Structural and Computational Biology, The Scripps Research Institute, La Jolla, CA 92037, USA.
10Ragon Institute of Massachusetts General Hospital, Massachusetts Institute of Technology, and Harvard University, Cambridge, MA 02139, USA.
↵†Corresponding author. Email: dsok{at}iavi.org (D.S.); jjardine{at}iavi.org (J.G.J.); burton{at}scripps.edu (D.R.B.)
↵* These authors contributed equally to this work.
Science 21 Aug 2020:
DOI: 10.1126/science.abc7520
Thomas F. Rogers
ORCID record for Thomas F. Rogers
Fangzhu Zhao
ORCID record for Fangzhu Zhao
Deli Huang
ORCID record for Deli Huang
Nathan Beutler
ORCID record for Nathan Beutler
Alison Burns
Wan-ting He
ORCID record for Wan-ting He
Oliver Limbo
ORCID record for Oliver Limbo
ORCID record for Chloe Smith
Ge Song
ORCID record for Ge Song
Jordan Woehl
ORCID record for Jordan Woehl
Linlin Yang
ORCID record for Linlin Yang
Robert K. Abbott
ORCID record for Robert K. Abbott
Sean Callaghan
Elijah Garcia
Jonathan Hurtado
ORCID record for Jonathan Hurtado
Mara Parren
Linghang Peng
Sydney Ramirez
ORCID record for Sydney Ramirez
James Ricketts
Michael J. Ricciardi
ORCID record for Michael J. Ricciardi
Stephen A. Rawlings
Nicholas C. Wu
ORCID record for Nicholas C. Wu
ORCID record for Meng Yuan
Davey M. Smith
ORCID record for Davey M. Smith
David Nemazee
ORCID record for David Nemazee
John R. Teijaro
ORCID record for John R. Teijaro
ORCID record for James E. Voss
Ian A. Wilson
ORCID record for Ian A. Wilson
Raiees Andrabi
Bryan Briney
ORCID record for Bryan Briney
Elise Landais
ORCID record for Elise Landais
Devin Sok
ORCID record for Devin Sok
For correspondence: dsok@iavi.org jjardine@iavi.org burton@scripps.edu
Joseph G. Jardine
ORCID record for Joseph G. Jardine
Dennis R. Burton
ORCID record for Dennis R. Burton
Protective neutralizing antibodies
Antibodies produced by survivors of coronavirus disease 2019 (COVID-19) may be leveraged to develop therapies. A first step is identifying neutralizing antibodies, which confer strong protection against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Rogers et al. used a high-throughput pipeline to isolate and characterize monoclonal antibodies from convalescent donors. Antibodies were selected for binding to the viral spike protein, which facilitates entry into host cells by binding to the angiotensin-converting enzyme 2 (ACE2) receptor. Most isolated antibodies bound to regions of the spike outside of the receptor binding domain (RBD); however, a larger proportion of the RBD-binding antibodies were neutralizing, with the most potent binding at a site that overlaps the ACE2 binding site. Two of the neutralizing antibodies were tested in Syrian hamsters and provided protection against SARS-CoV-2 infection.
Science, this issue p. 956
Countermeasures to prevent and treat coronavirus disease 2019 (COVID-19) are a global health priority. We enrolled a cohort of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)–recovered participants, developed neutralization assays to investigate antibody responses, adapted our high-throughput antibody generation pipeline to rapidly screen more than 1800 antibodies, and established an animal model to test protection. We isolated potent neutralizing antibodies (nAbs) to two epitopes on the receptor binding domain (RBD) and to distinct non-RBD epitopes on the spike (S) protein. As indicated by maintained weight and low lung viral titers in treated animals, the passive transfer of a nAb provides protection against disease in high-dose SARS-CoV-2 challenge in Syrian hamsters. The study suggests a role for nAbs in prophylaxis, and potentially therapy, of COVID-19. The nAbs also define protective epitopes to guide vaccine design.
The novel coronavirus disease 2019 (COVID-19) has had devastating global health consequences, and there is currently no cure or licensed vaccine. Neutralizing antibodies (nAbs) to the causative agent of the disease, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), represent potential prophylactic and therapeutic options and could help guide vaccine design. A nAb to another respiratory virus, respiratory syncytial virus (RSV), is in widespread clinical use prophylactically to protect vulnerable infants (1). Furthermore, nAbs prevent death from the emerging Ebola virus in macaques, even when given relatively late in infection, and thus have been proposed for use in outbreaks (2, 3). Generally, nAbs with outstanding potency (known as super-antibodies) (4) can be isolated by deeply mining antibody responses of a sampling of infected donors. Outstanding potency coupled with engineering to extend antibody half-life from weeks to many months brings down the effective costs of antibodies and suggests more opportunities for prophylactic intervention. At the same time, outstanding potency can permit antiviral therapeutic efficacy that is not observed for less potent antibodies (4). Here, we present the isolation of highly potent nAbs to SARS-CoV-2 and demonstrate their in vivo protective efficacy in a small animal model, suggesting their potential utility as a medical countermeasure.
To investigate the antibody response against SARS-CoV-2 and discover nAbs, we adapted our pipeline to rapidly isolate and characterize monoclonal antibodies (mAbs) from convalescent donors (Fig. 1). A cohort of previously swab-positive SARS-CoV-2 donors was recruited for peripheral blood mononuclear cell (PBMC) and plasma collection. In parallel, we developed both live replicating and pseudovirus neutralization assays using a HeLa-ACE2 (angiotensin-converting enzyme 2) cell line that gave robust and reproducible virus titers. Convalescent serum responses were evaluated for neutralization activity against SARS-CoV-1 and SARS-CoV-2, and eight donors were selected for mAb discovery. Single antigen-specific memory B cells were sorted, and their corresponding variable genes were recovered and cloned using a high-throughput production system that enabled antibody expression and characterization in under 2 weeks. Promising mAbs were advanced for further biophysical characterization and in vivo testing.
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Fig. 1 SARS-CoV-2 neutralizing antibody isolation strategy.
A natural infection cohort was established to collect plasma and PBMC samples from individuals who recovered from COVID-19. In parallel, functional assays were developed to rapidly screen plasma samples for SARS-CoV-2 neutralizing activity. SARS-CoV-2 recombinant surface proteins were also produced for use as baits in single-memory B cell sorting and downstream functional characterization of isolated mAbs. Finally, a Syrian hamster animal model was set up to evaluate mAb passive immunization and protection. The standard mAb isolation pipeline was optimized to facilitate high-throughput amplification, cloning, expression, and functional screening of hundreds of unpurified Ab heavy and light chain pairs isolated from each of several selected neutralizers in only 10 days. Selected pairs were scaled up to purify IgG for validation and characterization experiments. Potent neutralizing mAbs were selected to evaluate protection in the Syrian hamster model. HC, heavy chain; κC, kappa light chain; ΛC, lambda light chain; RT, reverse transcriptase.
Development of viral neutralization assays
Two platforms were established to evaluate plasma neutralization activity against SARS-CoV-2, one using replication-competent virus and another using pseudovirus (PSV). Vero-E6 cells were first used as target cells for neutralization assays, but this system was relatively insensitive at detecting replicating virus compared with a HeLa cell line that stably expressed the cell surface ACE2 receptor (fig. S1A). The HeLa-ACE2 target cells gave reproducible titers and were used for the remainder of the study. In certain critical instances, HeLa-ACE2 and Vero cells were compared.
The live replicating virus assay used the Washington strain of SARS-CoV-2, USA-WA1/2020 (BEI Resources NR-52281) and was optimized to a 384-well format to measure plaque formation. In parallel, a PSV assay was established for both SARS-CoV-1 and SARS-CoV-2 using murine leukemia virus–based PSV (MLV-PSV) (5). The assay used single-cycle infectious viral particles bearing a firefly luciferase reporter for high-throughput screening. Unlike MLV-PSV, which buds at the plasma membrane, coronaviruses assemble in the endoplasmic reticulum (ER)–Golgi intermediate compartment, so the C terminus of the SARS-CoV-1 spike (S) protein contains an ER retrieval signal (6). The alignment of SARS-CoV-1 and SARS-CoV-2 S proteins showed that this ER retrieval signal is conserved in SARS-CoV-2 (fig. S1B). To prepare high titers of infectious SARS-CoV-1 and SARS-CoV-2 PSV particles, various truncations of SARS-CoV-1 and SARS-CoV-2 S protein were expressed in which the ER retrieval signal was removed to improve exocytosis of the virus. Pseudovirion versions carrying SARS-CoV1-SΔ28 and SARS-CoV2-SΔ18S protein efficiently transduced ACE2-expressing target cells but not control HeLa or A549 cells (fig. S1C). Control VSV-G pseudotyped virions showed a similar transduction efficiency in all target cells. Luciferase expression in transduced cells proved to be proportional to viral titer over a wide range (fig. S1D).
Establishment of a SARS-CoV-2 cohort
In parallel to the development of neutralization assays, a cohort was established in San Diego, California, of 17 donors who had previously been infected with SARS-CoV-2 (Fig. 2A, fig. S2A, and table S1). The cohort was 47% female, and the average age was 50 years. Infection was determined by a positive SARS-CoV-2 polymerase chain reaction (PCR) test from a nasopharyngeal swab. All donors also had symptoms consistent with COVID-19, and disease severity ranged from mild to severe, including intubation in one case, although all donors recovered. Donor plasma were tested for binding to recombinant SARS-CoV-2 and SARS-CoV-1 S and receptor binding domain (RBD) proteins, for binding to cell surface–expressed spikes, and for neutralization in both live replicating virus and PSV assays [Fig. 2, B to D, and fig. S2B; three donors (CC6, CC12, and CC25) that are further discussed below are highlighted]. Binding titers to SARS-CoV-2 S protein varied considerably, reaching a half-maximal effective concentration (EC50) at serum dilutions of ~104, with titers against the RBD about an order of magnitude less. Titers against SARS-CoV-1 S protein were notably less than those for SARS-CoV-2 S protein, and titers against SARS-CoV-1 RBD were only detected in a small number of donors. Neutralizing titers in the PSV assay varied over a wide range for SARS-CoV-2 (Fig. 2D and fig. S2A) and were low or undetectable against SARS-CoV-1. RBD binding and PSV neutralization were notably well correlated (Fig. 2E). There was also a positive correlation between cell surface spike binding and live replicating virus neutralization (fig. S2C). The titers in the PSV assay and the replicating virus assay were largely similar (figs. S2 and S3). In most later measurements, the PSV assay was preferred owing to its higher throughput.
Fig. 2 COVID-19 cohort functional screening.
(A) Demographics of the University of California, San Diego (UCSD) COVID-19 cohort (CC) participants. CC plasma was tested for binding to SARS-CoV-1 and SARS-CoV-2 S proteins (B) and RBD subunits (C) by ELISA. Background binding of plasma to bovine serum albumin–coated plates is represented by a dotted line. OD405nm, optical density for wavelength of 405 nm. (D) Plasma was also tested for neutralization of pseudotyped (PSV) SARS-CoV-1 and SARS-CoV-2 virions. (E) Correlation between PSV SARS-CoV-2 neutralization and RBD subunit ELISA binding AUC (area under the curve). AUC was computed using Simpson's rule. The 95% confidence interval of the regression line is shown by the gray shaded area and was estimated by performing 1000 bootstrap resamplings. R2 (coefficient of determination) and P values of the regression are also indicated. CC participants from whom mAbs were isolated are specifically highlighted in blue (CC6), green (CC12), and pink (CC25).
Antibody isolation and preliminary functional screens for down-selection
Cryopreserved PBMCs from eight donors were stained for memory B cells markers (CD19+/IgG+; IgG, immunoglobulin G) and both AviTag biotinylated RBD and SARS-CoV-2 S antigen baits before single-cell sorting. S+ and S+/RBD+ memory B cells were present at an average frequency of 2.0 and 0.36%, respectively, across the eight donors (fig. S4A). In total, 3160 antigen-positive (Ag+) memory B cells were sorted to rescue native heavy and light chain pairs for mAb production and validation (fig. S4B). A total of 2045 antibodies were cloned and expressed, which represents, on average, a 65% PCR recovery of paired variable genes and >86% estimated recovery of fully functional cloned genes (fig. S4C). The bulk-transformed ligation products for both the heavy chain and light chain were transfected and tested for binding to RBD and S protein and for neutralization in the SARS-CoV-2 PSV assay using HeLa-ACE2 target cells (fig. S5).
The majority of transfected pairs (92%) resulted in IgG expression. Of these, 43% showed binding only to S protein, while 5.9% bound to both S and RBD proteins and 0.1% bound only to RBD. The supernatants were also screened for binding to an unrelated HIV antigen (BG505 SOSIP) to eliminate nonspecific or polyreactive supernatants. The supernatants were next evaluated for neutralization activity using SARS-CoV-2 and SARS-CoV-1 pseudoviruses. A small proportion of the binding antibodies showed neutralization activity, and, unexpectedly, that activity was equally distributed between RBD+/S+ and S+-only binders, despite a much larger number of S+-only binding supernatants, as exemplified by the three donors, CC6, CC12, and CC25 (Fig. 3A). These data indicate that viral infection generates a strong response against the non-RBD regions of the S protein, but only a small proportion of that response is neutralizing. In contrast, there are fewer RBD-binding antibodies, but a larger proportion of these neutralize SARS-CoV-2 pseudovirus. Antibodies that tested positive for neutralization in the high-throughput screening were sequence-confirmed and advanced for large-scale expression for additional characterization.
Fig. 3 Antibody isolation and functional screening for SARS-CoV antigen binding and neutralization.
(A) Antibody down-selection process from three donors, presented as bubble plots. The areas of the bubbles for each donor are sized according to the number of antibodies (n) that were cloned and transfected, then scaled according to the number that were positive in subsequent assays. All antibodies that expressed at measurable levels were tested for binding to S protein and RBD to determine their specificity and then screened for neutralization. (B) VH gene distribution of down-selected mAbs. (C) Heavy chain CDR3 lengths of down-selected mAbs. Antibodies in (B) and (C) are colored according to their respective clonal lineages. (D) Mutation frequency of down-selected mAb lineages. Bubble position represents the mean mutation frequency for each lineage, with a bubble area that is proportional to lineage size. LC, light chain; nt, nucleotides.
Thirty-three antibodies were prioritized for in-depth characterization from the three donors, CC6, CC12, and CC25. Within that subset, we identified 25 distinct lineages, with 23 containing a single member (table S2). VH1 and VH3 gene families were notably prominent in these Abs, and there was a diversity of CDR3 lengths (Fig. 3, B and C). There was one prominent example of a clonally expanded lineage, with eight recovered clonal members that averaged 4.3 and 2.8% mutations from germline at the nucleotide level in the heavy chain and light chain, respectively (Fig. 3D). The remaining clones were relatively unmutated, averaging just above 1% mutation at the nucleotide level, suggesting that these antibodies were primed by the ongoing COVID infection and likely not recalled from a previous endemic human coronavirus exposure. All antibodies that were expressed at scale were evaluated in standard enzyme-linked immunosorbent assay (ELISA)–based polyreactivity assays with solubilized Chinese hamster ovary (CHO) membrane preparations, single-stranded DNA, and insulin (7, 8), and none were polyreactive (fig. S6).
Functional activity of down-selected antibodies
The antibody hits that were identified in the high-throughput screening were next evaluated for epitope specificity by biolayer interferometry using S and RBD proteins as capture antigens. The antigens were captured on anti-HIS biosensors before addition of saturating concentrations (100 μg/ml) of antibodies that were then followed by competing antibodies at a lower concentration (25 μg/ml). Accordingly, only antibodies that bind to a noncompeting site would be detected in the assay. Among the antibodies evaluated, the results reveal three epitope bins for RBD (designated RBD-A, RBD-B, and RBD-C) and three epitope bins for the S protein (designated S-A, S-B, and S-C) (Fig. 4A and fig. S7). The mAb CC12.19 appears to compete with antibodies targeting two different epitopes, RBD-B and S-A (fig. S7), which might indicate that this mAb targets an epitope spanning RBD-B and S-A. To evaluate epitope specificities further, we next assessed binding of the antibodies to extended RBD constructs with subdomains (SD) 1 and 2, including the independently folding RBD-SD1 and RBD-SD1-2 and the N-terminal domain (NTD) (Fig. 4B and fig. S8, A and B). None of the antibodies showed binding to the NTD. CC12.19 binds to all other constructs, which supports the epitope binning data described in Fig. 4A. The other antibodies grouped in the S-A epitope bin that compete with CC12.19 either showed no binding to RBD or RBD-SD constructs (CC12.20 and CC12.21) or showed binding to RBD-SD1 and RBD-SD1-2 but not RBD (CC12.23). These data suggest two competing epitopes within the S-A epitope bin: one that is confined to the non-RBD region of the S protein, and one that includes some element of RBD-SD1-2. This interpretation will require further investigation by structural studies.
Fig. 4 Antibody functional activity by epitope specificities.
Monoclonal antibody epitope binning was completed using RBD and SARS-CoV-2 S protein as target antigens. (A) A total of three noncompeting epitopes for RBD (RBD-A, RBD-B, and RBD-C) and three noncompeting epitopes for S (S-A, S-B, and S-C) were identified. (B) MAbs were evaluated for binding to different target antigens (S, NTD, RBD, RBD-SD1, and RBD-SD1-2) by ELISA and apparent EC50 values are reported in micrograms per milliliter. (C) MAbs were evaluated for neutralization of SARS-CoV-2 pseudovirus using HeLa-ACE2 target cells. Antibodies are grouped according to epitope specificities, and neutralization IC50 values are reported in micrograms per milliliter. (D) The MNP is reported for each mAb and grouped by epitope specificity. MAbs were mixed with (E) S or (F) RBD protein and measured for binding to HeLa-ACE2 target cells as a measure of competition to the cell surface ACE-2 receptor. (G) mAb neutralization potencies (IC50) are plotted as a function of dissociation constants (KD) measured by SPR to RBD target antigen.
We next evaluated the mAbs for neutralization activity against SARS-CoV-2 and SARS-CoV-1 pseudoviruses. The neutralization half-maximal inhibitory concentration (IC50) potencies of these antibodies are shown in Fig. 4C, and their associated maximum neutralization plateaus (MNPs) are shown in Fig. 4D. A comparison of neutralization potencies between pseudovirus (fig. S8C) and live replicating virus (fig. S8D) is also included. The most potent neutralizing antibodies were those directed to RBD-A epitope, including two antibodies, CC6.29 and CC6.30, that neutralize SARS-CoV-2 pseudovirus with an IC50 of 2 ng/ml and 1 ng/ml, respectively (Fig. 4C). In comparison, antibodies directed to RBD-B tended to have a higher IC50 and many plateaued below 100% neutralization. Despite this trend, CC6.33 is directed against RBD-B and showed complete neutralization of SARS-CoV-2 with an IC50 of 39 ng/ml and also neutralized SARS-CoV-1 with an IC50 of 162 ng/ml. CC6.33 was the only antibody that showed potent neutralization of both pseudoviruses. The antibodies that do not bind to RBD and are directed to non-RBD epitopes on the S protein all showed poor neutralization potencies and MNPs well below 100%.
To evaluate whether the RBD-A epitope might span the ACE2 binding site, we next performed cell surface competition experiments. Antibodies were premixed with biotinylated S (Fig. 4E) or RBD (Fig. 4F) proteins at a 4:1 molar ratio of antibodies to target antigen. The mixture was then incubated with the HeLa-ACE2 cell line and the percent competition against ACE2 receptor was recorded by comparing percent binding of the target antigen with and without antibody present (fig. S8E). The antibodies targeting the RBD-A epitope competed best against the ACE2 receptor, and the neutralization IC50 correlated well with the percent competition for ACE2 receptor binding for both S protein (Fig. 4E) and RBD (Fig. 4F). We also assessed the affinity of all RBD-specific antibodies to soluble RBD by surface plasmon resonance (SPR) and found a poor correlation between affinity and neutralization potency (Fig. 4G and fig. S9). However, the correlation is higher when limited to antibodies targeting the RBD-A epitope. The lack of a correlation between RBD binding and neutralization for mAbs contrasts with the strong correlation described earlier for serum RBD binding and neutralization. Overall, the data highlight epitope RBD-A as the preferred target for eliciting neutralizing antibodies and suggest that corresponding increases in affinity of mAbs to RBD-A will likely result in corresponding increases in neutralization potency.
SARS-CoV-2 has shown some propensity for mutation as it has circulated worldwide, as evidenced, for example, in the emergence of the D614G variant (9). We investigated the activity of five nAbs against six viral variants that have been reported. The three sera studied above (CC6, CC12, and CC25) neutralized all the variants (fig. S10A). All five nAbs neutralized the D614G variant. However, one variant with a mutation in the ACE2 binding site [Gly476→Ser (G476S)] did show effectively complete resistance to one of the nAbs, and another variant (V367F) showed a 10-fold higher IC50 than the WA1 strain (fig. S10B).
Passive transfer of neutralizing antibodies and SARS-CoV-2 challenge in Syrian hamsters
To investigate the relationship between in vitro neutralization and protection in vivo against SARS-CoV-2, we selected two mAbs for passive transfer and challenge experiments in a Syrian hamster animal model on the basis of a summary of the nAb data (table S3 and fig. S11). The experimental design for the passive transfer study is shown in Fig. 5A. In the first experiment, we tested nAb CC12.1, which targets the RBD-A epitope and has an in vitro IC50 neutralization of 0.019 μg/ml against pseudovirus, and in the second experiment, we tested nAb C12.23, which targets the S-B epitope with an IC50 neutralization of 22 μg/ml. In both experiments, an unrelated antibody to dengue virus, Den3, was used as a control. The anti-SARS-CoV-2 nAbs were delivered at five different concentrations to evaluate dose-dependent protection, starting at 2 mg per animal (average: 16.5 mg/kg) at the highest dose and 8 μg per animal at the lowest dose. The Den3 control antibody was delivered at a single dose of 2 mg per animal. Sera were collected from each animal 12 hours after intraperitoneal infusion of the antibody, and all animals were subsequently challenged with a dose of 1 × 106 plaque forming units (PFU) of SARS-CoV-2 (USA-WA1/2020) by intranasal administration 12 hours after antibody infusion (Fig. 5A).
Fig. 5 A potent SARS-CoV-2 RBD-specific neutralizing mAb protects against weight loss and lung viral replication in Syrian hamsters.
(A) SARS-CoV-2–specific human neutralizing mAb CC12.1 isolated from natural infection was administered at a starting dose of 2 mg per animal (average: 16.5 mg/kg) and subsequent serial fourfold dilutions. Control animals received 2 mg of Den3. Each group of six animals was challenged intranasally (i.n.) 12 hours after infusion with 1 × 106 PFU of SARS-CoV-2. Serum was collected at the time of challenge (day 0), and animal weight was monitored as an indicator of disease progression. On day 5, lung tissue was collected for viral burden assessment. (B) Percent weight change was calculated from day 0 for all animals. (C) Viral load, as assessed by nucleocapsid RNA quantitative polymerase chain reaction (qPCR) from lung tissue at day 5 after infection. (D) Serum titers of the passively administered mAb, as assessed by ELISA at the time of challenge [12 hours after intraperitoneal (i.p.) administration]. Correlation analyses with 95% confidence intervals indicated by the gray shaded area.
Syrian hamsters typically clear virus within 1 week after SARS-CoV-1 infection, and observations made in that model system determined the strategy adopted here (10). The hamsters were weighed daily as a measure of disease due to infection. Lung tissues were collected to measure viral load on day 5. A data summary is presented in Fig. 5B and fig. S12A for animals that received CC12.1, which targets the RBD-A epitope. The control animals that received Den3 lost, on average, 13.6% of body weight by day 5 after virus challenge. In comparison, the animals that received the neutralizing RBD-A antibody at a dose of 2 mg (average: 16.5 mg/kg) or 500 μg (average: 4.2 mg/kg) exhibited no weight loss. However, animals that received a dose of 125 μg (average: 0.9 mg/kg) had an average body weight loss of 8%, while animals that received a dose of 31 μg/ml (average: 0.2 mg/kg) and 8 μg/ml (average: 0.06 mg/kg) lost 15.8 and 16.7% of body weight, respectively. Although these animals showed a trend for greater weight loss than did control animals, this trend did not achieve statistical significance (table S4). Given concerns about antibody-mediated enhanced disease in SARS-CoV-2 infection, this observation merits further attention using larger animal group sizes. The weight loss data are further corroborated by quantification of lung viral load measured by real-time PCR (Fig. 5C), which showed a moderate correlation to weight loss. The data indicate comparable viral loads between the three higher doses (2 mg, 500 μg, and 125 μg) of nAbs. In contrast, equivalent viral loads were observed between the control group receiving Den3 and the low-dose groups receiving 31 or 8 μg of nAb. In contrast to the nAb to RBD-A, the less potent and incompletely neutralizing antibody to the S-B epitope showed no evidence of protection at any concentration when compared with control animals (fig. S12B).
To determine the antibody serum concentrations that may be required for protection against disease from SARS-CoV-2 infection, we also measured the antibody serum concentrations just before intranasal virus challenge (Fig. 5D). The data highlight that an antibody serum concentration of ~22 μg/ml of nAb (1160 × PSV neutralization IC50) enables full protection and a serum concentration of 12 μg/ml (630 × PSV neutralization IC50) is adequate for a 50% reduction in disease, as measured by weight loss. The effective antibody concentration required at the site of infection to protect from disease remains to be determined. Sterilizing immunity at serum concentrations that represent a large multiplier of the in vitro neutralizing IC50 is observed for many viruses (11).
Using a high-throughput rapid system for antibody discovery, we isolated more than 1000 mAbs from three convalescent donors by memory B cell selection using SARS-CoV-2 S or RBD recombinant proteins. About half of the mAbs isolated could be expressed, and they also bound effectively to S and/or RBD proteins. Only a small fraction of these Abs were neutralizing, which highlights the value of deep mining of responses to access the most potent Abs (4).
A range of nAbs were isolated to different sites on the S protein. The most potent Abs, reaching single-digit nanogram per milliliter IC50 values in PSV assays, are targeted to a site that, judged by competition studies, overlaps the ACE2 binding site. Only one of the Abs, directed to RBD-B, neutralized SARS-CoV-1 PSV, as may be anticipated given the differences in ACE2 contact residues between the two viruses (fig. S13) and given that the selections were performed with SARS-CoV-2 target proteins. Abs that are directed to the RBD but not competitive with soluble ACE2 (although they may be competitive in terms of an array of membrane-bound ACE2 molecules interacting with an array of S proteins on a virion) are generally less potent neutralizers and tend to show incomplete neutralization, plateauing well below 100% neutralization. The one exception is the cross-reactive RBD-B antibody, mentioned above. Similarly low potency and incomplete neutralization are observed for Abs to the S protein that are not reactive with recombinant RBD. The cause(s) of these incomplete neutralization phenomena is unclear but presumably originates in some S protein heterogeneity that is either glycan, cleavage, or conformationally based. Regardless, the RBD-A nAbs that directly compete with ACE2 are clearly the most preferred for prophylactic and therapeutic applications and as reagents to define nAb epitopes for vaccine design. Note that, even for a small sampling of naturally occurring viral variants, two were identified that showed notable resistance to individual potent nAbs to the WA1 strain, which suggests that neutralization resistance will need to be considered in planning for clinical applications of nAbs. Cocktails of nAbs may be required.
In terms of nAbs as passive reagents, the efficacy of a potent anti-RBD nAb in vivo in Syrian hamsters is promising in view of the positive attributes of this animal model (12) and suggests that human studies are merited. Nevertheless, as for any animal model, there are many limitations, including, in the context of antibody protection, differences in effector cells and Fc receptors between humans and hamsters. The failure of the non-RBD S-protein nAb to protect in the animal model is consistent with its lower potency and, likely most importantly, its inability to fully neutralize challenge virus. In the context of human studies, the following antibody engineering goals could be considered: improving the potency of protective nAbs by enhancing binding affinity to the identified RBD epitope, improving half-life, and reducing Fc receptor binding to minimize potential antibody-dependent enhancement (ADE) effects if they are identified. As observed for heterologous B cell responses against different serotypes of flavivirus infection, there is a possibility, but no current experimental evidence, that subtherapeutic vaccine serum responses or subtherapeutic nAb titers could potentially exacerbate future coronavirus infection disease burden by expanding the viral replication and/or cell tropism of the virus. If ADE is found for SARS-CoV-2 and operates at subneutralizing concentrations of neutralizing antibodies, as it can for dengue virus (13), then it would be important, from a vaccine standpoint, to carefully define the full range of nAb epitopes on the S protein, as we have begun to do here. From a passive antibody standpoint, it would be important to maintain high nAb concentrations or appropriately engineer nAbs.
The nAbs described here have very low somatic hypermutation (SHM), typically only one or two mutations in the VH gene and one or two in the VL gene. Such low SHM may be associated with the isolation of the nAbs relatively soon after infection, perhaps before affinity-maturation has progressed. Low SHM has also been described for potent nAbs to Ebola virus, RSV, Middle East respiratory syndrome coronavirus, and yellow fever virus (14–17) and may indicate that the human naïve repertoire is often sufficiently diverse to respond effectively to many pathogens with little mutation. Of course, nAb efficacy and titer may increase over time, as described for other viruses, and it will be interesting to see if even more potent nAbs to SARS-CoV-2 evolve in our donors in the future.
What do our results suggest for SARS-CoV-2 vaccine design? First, they suggest a focus on the RBD—strong nAb responses have indeed been demonstrated by immunizing mice with a multivalent presentation of RBD (18). The strong preponderance of non-neutralizing antibodies and the very few nAbs to S protein that we isolated could arise for a number of reasons, including the following: (i) The recombinant S protein that we used to select B cells is a poor representation of the native spike on virions. In other words, there may be many nAbs to the S protein, but we failed to isolate them because of the selecting antigen. (ii) The recombinant S protein that we used is close to native, but non-neutralizing antibodies bind to sites on the S protein that do not interfere with viral entry. (iii) The S protein in natural infection disassembles readily, generating a strong Ab response to viral debris that is non-neutralizing, because the antibodies recognize protein surfaces that are not exposed on the native spike. The availability of both neutralizing and non-neutralizing antibodies generated in this study will facilitate evaluation of S protein immunogens for presentation of neutralizing and non-neutralizing epitopes and will promote effective vaccine design. The design of an immunogen that improves on the quality of nAbs elicited by natural infection may well emerge as an important goal of vaccine efforts (19).
In summary, we describe the very rapid generation of neutralizing antibodies to a newly emerged pathogen. The antibodies can find clinical application and will aid in vaccine design.
science.sciencemag.org/content/369/6506/956/suppl/DC1
Figs. S1 to S13
Tables S1 to S4
References (20–27)
MDAR Reproducibility Checklist
View/request a protocol for this paper from Bio-protocol.
This is an open-access article distributed under the terms of the Creative Commons Attribution license, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
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Acknowledgments: We thank T. Gilman, A. Salazar, and Biosero for their contribution to high-throughput pipeline generation. We also thank B. Graham and J. S. McLellan for provision of materials. We thank L. Walker for valuable manuscript discussions. We thank all the COVID-19 cohort participants for donating samples. Funding: This work was supported by the NIH CHAVD (UM1 AI144462 to I.A.W., B.B., D.S., and D.R.B.), R01 (AI132317 and AI073148 to D.N.), and K99 (AI145762 to R.K.A.) awards; the IAVI Neutralizing Antibody Center; and the Bill and Melinda Gates Foundation (OPP 1170236 to I.A.W. and D.R.B., OPP 1183956 to J.E.V., and OPP1196345/ INV-008813 to D.S. and D.R.B.). This work was also supported by the John and Mary Tu Foundation and the Pendleton Trust. Author contributions: T.F.R., E.L., D.S, J.G.J., and D.R.B. conceived of and designed the study. T.F.R., N.B., E.G., M.P., S.R., S.A.R., and D.M.S. recruited donors and collected and processed plasma and PBMC samples. F.Z., A.B., L.Y., M.P., and G.S. performed ELISA and cell binding assays. F.Z., D.H., N.B., L.Y., M.P., L.P., J.E.V., and J.R. developed and performed neutralization experiments. F.Z., A.B., O.L., C.S., J.W., R.K.A., E.L., D.S., and J.G.J. performed antibody isolation, cloning, expression, and competition experiments. W.-t.H., G.S., S.C., R.A., N.C.W., M.Y., and I.A.W. generated recombinant antigens. T.F.R., L.Y., M.J.R., and J.R.T. carried out animal studies and qPCR viral load measurements. T.F.R., F.Z., D.H., N.B., G.S., J.W., J.H., M.J.R., D.N., J.R.T., R.A., B.B., E.L., D.S., J.G.J., and D.R.B. designed the experiments and analyzed the data. E.L., D.S., J.G.J., and D.R.B. wrote the paper, and all authors reviewed and edited the paper. Competing interests: D.R.B., D.H., J.G.J., E.L., T.F.R., D.S., and F.Z. are listed as inventors on pending patent applications describing the SARS-CoV-2 antibodies. D.R.B. is a consultant for IAVI. All other authors have no competing interests to declare. Data and materials availability: Antibody sequences have been deposited in GenBank under accession numbers MT594038 to MT594103. Antibody plasmids are available from D.R.B. under an MTA from the Scripps Research Institute. This work is licensed under a Creative Commons Attribution 4.0 International (CC BY 4.0) license, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. To view a copy of this license, visit https://creativecommons.org/licenses/by/4.0/. This license does not apply to figures/photos/artwork or other content included in the article that is credited to a third party; obtain authorization from the rights holder before using such material.
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By Thomas F. Rogers, Fangzhu Zhao, Deli Huang, Nathan Beutler, Alison Burns, Wan-ting He, Oliver Limbo, Chloe Smith, Ge Song, Jordan Woehl, Linlin Yang, Robert K. Abbott, Sean Callaghan, Elijah Garcia, Jonathan Hurtado, Mara Parren, Linghang Peng, Sydney Ramirez, James Ricketts, Michael J. Ricciardi, Stephen A. Rawlings, Nicholas C. Wu, Meng Yuan, Davey M. Smith, David Nemazee, John R. Teijaro, James E. Voss, Ian A. Wilson, Raiees Andrabi, Bryan Briney, Elise Landais, Devin Sok, Joseph G. Jardine, Dennis R. Burton
Science 21 Aug 2020 : 956-963
Passive transfer of a neutralizing antibody provides protection against disease in high-dose SARS-CoV-2 challenge in Syrian hamsters.
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Hobart vs Launceston. Cascade vs Boags. Mount Wellington vs Cradle Mountain. Gagebrook bogans vs Ravenswood bogans. Tasmania is often spoken about by locals as being a very parochial state. We have the North/South divide, with each half of the state engaged in a long running battle about who has the best stuff. In the new game of Tasmanian whisky, the cards are very much stacked on the side of the South, with eight of the nine distilleries in the state residing there.
However, the North cannot be swept off the board that easily, as it has a very impressive golden ace up its sleeve. Hellyers Road, located in the North-West city of Burnie, has officially launched its 12 Year Old whisky, making it the first distillery in Australia to release an expression of this age. In 2012 the company released a 10 Year Old single malt which showed promise of greatness, and now two years later that potential is being realised. Pleasingly the extra years spent sleeping in oak have helped smooth out the edges without compromising the creamy, buttery flavours unique to Hellyers Road.
Compared to the white wine colour palate of the Hellyers Road Original, the longer time in the barrels has imparted a rich golden hue to the 12. The nose opens with vanilla from the American bourbon oak, followed by the creamy nuttiness of macadamias, cashews and almonds. There are also elements of candied citrus peel and melted butter to be found. The overall effect is of vanilla cupcakes coated in orange and poppy seed icing.
On the palate the 12 is smoother and yet more complex than other Hellyers Road expressions, with delicate honeyed undertones that are reminiscent of the lightly burnt sugar on top of a crème brulee. Complementing the sweetness are subtle herbal notes and spice that bring to mind the leaves of the Tasmanian native pepper berry bush. The finish is light and imparts a soft warmth to the back of the throat.
When we asked Hellyers Road head distiller Mark Littler if the 12 was everything he intended it to be, his simple reply was that "it's more". Two years may not seem a long time to us, but to this whisky that short period is incredibly significant and adds a high class edge of silk into the mix. The 12 year old is the defining expression of Hellyers Road, and an exciting move forward for Tasmanian whisky. As Northern boys we're proud to say that the golden ace has been played with style in our end of the state, and taken the game to a whole new level.
Wow, sounds like an excellent whisky! I know absolutely nothing about Australian whisky – and I'm not sure I be ever even seen one here in Canada. As a single malt, I'm guessing that barley is the main grain used, but do other grains make an appearance in any of your whiskies? I'm interested because Canadian whiskies often use corn, wheat, and especially rye. Also, what abv would a 12 year old like this one be bottled at?
It's a great one – keep an eye out for it! I'm confident it will be unlike any other whisky you've tried! Mostly all Tasmanian whiskies use barley, but there is one, called Belgrove which is 100% rye! The percentage varies, but it is often 46% ABV (which is the magic, non-chill filtered kind of percentage). I'm glad you liked the review!
Aha! Excellent! Have you ever tried any other Hellyers Road expressions? I certainly appreciated the 12 Year Old better when I had a bit of context from younger expressions. Anyway, I look forward to hearing your thoughts.


text:
Hess Benedict Road before and after AWA tree planting event.
We love looking back at our progress!
AWA was established as a 501 C 3 tax exempt non- profit organization in 2002.
Significant relationships with local government, particularly Washington township where AWA is now a line item in the town budget.
AWA local office established at 32 W Main Street, Waynesboro PA.
Creating new partnerships with local organizations like Renfrew Institute, local farms and municipalities and created a connection with Waynesboro School District.
Built a highly visible rain garden at the local farmers market, Paul's Market, to filter the runoff from the parking lot and part of the roadway.
Accomplished significant riparian planting projects at farms and two large landholders along the Antietam Creek in PA.
Planting over 5,000 trees.
Conducted an assessment of the East Branch Antietam Creek, resulting in recommendations to preserve and correct problems in the watershed.
Established and currently maintaining a scientifically rigorous stream monitoring program, in partnership with the Renfrew Institute, to gather data and use as an educational resource for the community.
Proceeded with plans for a major project of preservation and educational value on WASD property, an outcome of the East Branch assessment.
Adopted a State Forest road - Rattlesnake Road- and maintain an active cleanup of this road throughout the year and a yearly cleanup project.
Participated annually in community outreach events including the Appalachian
Trail Community Festival in June, Waynesboro's Market Day in October, Renfrew
Institute's Earth Celebration Day in April and the Blue Ridge Summit Library Ice
Cream Social in July.
Designed and published a brochure for general education on riparian planting in the PA watershed. Established a website, designed and published a brochure about us, AWA and created a display board for use at the public events.
In 2014 President Carl Grove built a Pinball Game that is used to demonstrate ways of keeping
pollution from entering waterways. It is used at these events.
Developed and distributed over 500 pamphlets on the importance of turning in
unused or outdated over the counter and prescription drugs instead of flushing
them or throwing them in the trash. Drugs can be turned in at the Waynesboro
Police Station Lobby or some pharmacies.
With the bat population in danger, AWA is excited to show off our new project building and setting up bat boxes in Antietam Meadows park.
Duck Derbies
With the generous support of community business we held two Duck Derbies to
raise operational funds.
Applied for and received grants for over $200,000. The grants were used
for stream restoration projects, educational materials and a contract with
RETTEW, Inc. to complete a study of the entire West Branch of the Antietam that
reported conditions and made recommendations to correct deficiencies.
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Adam Macleod was on the main road to the Skye Bridge when he was forced to swerve to avoid a potentially deadly smash.
This comes as former crash survivor, Sharon Anslow, has started the 'Keep Left' campaign after she was injured in a car incident involving a tourist in Skye last year.
The clip begins with Adam driving back home to Kyle of Lochalsh, Ross-Shire, on the A87 with his car radio playing in the background.
He can be seen crossing a bridge and in the corner of the shot, the roof of a car can be seen coming towards him on his side of the road.
As he comes round the corner, a black Skoda is seen directly in front of him.
Luckily for civil servant Adam, there were no other cars on the right hand side of the road, so he was able to quickly swerve his car over to the other side.
The 19-plate Skoda, which is thought to be a hire car, manages to pass without a collision as Adam gets himself back onto the left hand side of the road.
He can be heard saying "f****** choob" just before the clip ends.
Adam's post has since gained more than 1,500 and 300 comments from shocked Facebook users.
George MacInnes said: "If he swerved to avoid you at the same time as you did, then there would have been a crash with you on the wrong side of the road.
He said: "My heart certainly jumped a beat, hit the brakes, but they didn't move at all so I took the split second decision to swerve.
He also explained how incidents like this are becoming more common in the area.
He commented: "It's the first time that I have witnessed it, but I know of incidents within a few hundred yards of this that have happened in the last year.
and for hire car companies to place "keep left" stickers inside their vehicles.
Sharon said this was a common problem across the Highlands that needed resolving.
Speaking to a local broadcaster, she said: ""So many people contacted me after my accident saying: 'It happened to us'. Even earlier that day of my accident it had happened to somebody else.


Huge second quarter puts the game out of reach.
The Howard Bison thrilled fans with a win over UNLV and even thrilled some with a loss to Kent State, but there were no thrills to be had in Richmond. No. 6 Richmond was up 14-0 after the first quarter and cruised to a 68-21 win at Robins Stadium.
Howard (1-2) was held to 91 yards rushing and 197 passing yards.
Richmond outscored Howard 34-7 in the second quarter which all but put the game out of reach.
Caylin Newton did show a sparkle in the third quarter breaking away for a 17-yard TD, but there would be no comeback on Saturday afternoon.
Newton found Jequez Ezzard in the fourth for a 28-yard touchdown pass which would be the final touchdown of the game.
Richmond (2-1), finished the day with 622 total yards and ten touchdowns.
Howard hosts Bethune-Cookman in their home opener next Saturday, kickoff is slated for 1 p.m.


Submit your wine(s) to be tasted by Master Sommelier Cameron Douglas for only $60+GST per wine. Simply complete the form below for each wine.
Cameron will taste all wines submitted and score them. We will then email you Cameron's tasting notes and score for each wine. You may use these notes for your own marketing purposes.
You will then have the opportunity to feature your wine(s) in the corresponding 2019 issue of The Shout NZ Hospitality Business for $395+GST per wine. This will be part of a varietal feature written by Cameron and includes a high resolution bottle shot as well as Cameron's tasting notes, score and your contact information.
Deadline for submissions: Friday 5th April (wine to be received by C.O.B please).
© 2018 The Intermedia Ad Centre. All Rights Reserved.


text:
Home › Foxe's Book of Martyrs
Foxe's Book of Martyrs
Foxe, John | Revell
This Christian classic tells the stories of brave men and women who were martyred for their faith in the fourteenth to sixteenth centuries. They faced torture and martyrdom rather than deny their vision of truth and of God.
Beginning with Jesus Christ, this exceptional historical record traces the roots of religious persecution of those who followed Him. It examines the heroic lives of great men and women such as John Hus, John Wycliffe, William Tyndale, Anne Askew, Lady Jane Grey and Martin Luther. The faith of these early martyrs has stood, and will stand forever as a beacon to the generation that follows.
''After the Bible itself, no book so profoundly influenced early Protestant sentiment as the Book of Martyrs. Even in our time it is still a living force. More than a record of persecution, it is an arsenal of controversy, a storehouse of romance, and a source of edification.''
- James Miller Dodds, English Prose
Church History 101: The Highlights of Twenty Centuries [Paperback]
In the Year of Our Lord: Reflections on Twenty Centuries of Church History [Hardcover]
Meet the Puritans: With A Guide to Modern Reprints [Hardcover]
Reformation Women: Sixteenth-Century Figures Who Shaped Christianity's Rebirth [Paperback]


Auto restoration, paint sealants, paint repair, paintless dent removal, undercoatings, and accessories. Located in Akron, Ohio.
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Locally owned and operated. transmission repair and service, engine repair and swaps.
Car windshield and auto glass repair specialists in St. Paul and Minneapolis repair windshields and car windows. Providers of auto glass repair and home and business window pane replacement. Windshield repair services are insurance approved.
A Toronto premier towing service. Covering all areas of the GTA, reliable emergency road services are one call away.
Blossom Collision Center is an Indianapolis auto body repair & paint shop. We provide complete service on all makes & models. Bring your vehicle in for a free estimate today.
BMW Concord Collision is an auto body repair shop. Whether you need a quick touch-up or recently got in an accident, we will provide professional service to bring your BMW back to perfect shape.
Experts in car audio repair. Nationwide repair since 1972 on AM/FM, cassette, 8 Track, CD, and speakers. Repairing foreign and domestic models from 1955 and up.
A clear bra company in Cincinnati, Ohio whose services include installation of 3M Scotchguard and Xpel Ultimate clear bra paint films.
Full service truck, SUV and car services shop offers wide range of affordable car repairs, including brake inspection/brake repair, new shocks and struts and batteries and more: also provides fast truck, SUV and car maintenance, including oil lube change.
Offers extended warranty for new and used vehicles in US and Canada.
Provides BMW service and maintenance for all makes and models of BMW cars in the Bay Area. Providing BMW service for over 20 years.
Scrap car yard based in Bolton. Metro Salvage offer nationwide vehicle pickups and scrap vehicles of all types.
Towing Riders provide 24/7 quality towing service in Dallas, TX. we can take care of any emergency situation for you.
Provides key automotive maintenance and tire services in Maryland and Pennsylvania.
They have been shining your vehicles for nearly 75 years. Car detailing is Ming Shine Edmonton's specialty. We can also deal with your car's paint scratches & dents with quick small damage repair services.


text:
Car Reviews & Research
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Motor Illustrated
News Lamborghini Lambo V12 Vision Gran Turismo Revealed
Lamborghini Lambo V12 Vision Gran Turismo Revealed
Michel Deslauriers
The Lamborghini Lambo V12 Vision Gran Turismo is the latest downloadable supercar for the Gran Turismo racing video game on the PS4 console.
Lamborghini Lambo V12 Vision Gran Turismo designed for the Gran Turismo Sport racing game on the PS4 console
Said to be equipped with the Sián FKP 37's 807-horsepower hybrid powertrain
Single-seater supercar downloadable starting in the spring of 2020.
An all-new supercar will be available to download in the popular Sony Playstation title Gran Turismo Sport, and this time around, it comes from an Italian manufacturer. The Lamborghini Lambo V12 Vision Gran Turismo is a virtual concept car that actually exists, sort of.
The single-seater Lambo V12 is said to use the Lamborghini Sián FKP 37's powertrain, which consists of a naturally aspirated, 6.5-litre V12 engine and a 48-volt electric motor. Combined output for the Sián is rated at 807 horsepower (819 PS, 602 kW), allowing for a 0-to-100 km/h time of 2.8 seconds and a top speed in excess of 350 km/h. No performance numbers have been announced for the Lambo V12 Vision GT.
Jaguar Designs Virtual Vision Gran Turismo Coupé
Despite being a "virtual" car designed for a video game, Lamborghini presented a life-size model in Monte Carlo, just ahead of the World Finals of the 2019 FIA Certified Gran Turismo Championships. Yes, we're talking about a racing game that has the FIA's stamp of approval.
"Lamborghini is a very young brand, and this is why we are here today to present our newest virtual vision in the form of a real model, with a highly futuristic and cool design to be enjoyed by the young generation of racing game and super sports car enthusiasts," said Stefano Domenicali Chairman and CEO of Automobili Lamborghini S.p.A.
The car's exterior design is wild, aggressive and simply stunning. It looks like a cross between the Batmobile, a fighter jet and a giant venomous insect. Styling highlights include the magnificent golden yellow paint, the huge wheels, the front fenders disconnected from the body and the Y-shaped taillight that extends up into the rear wing.
"The Lambo V12 Vision Gran Turismo is created to provide the ultimate virtual car for young fans and gamers who are ultra-enthusiastic about Lamborghini and its futuristic aspirations," stated Mitja Borkert, Head of Lamborghini Centro Stile—the automaker's design studio in Sant'Agata Bolognese.
Obviously, there are no plans to produce the Lamborghini Lambo V12 Vision Gran Turismo concept. However, players of Gran Turismo Sport on the PS4 console will be able to download the car and enjoy it starting in spring 2020.
Lamborghini Lambo V12 Vision Gran Turismo
Lamborghini Vision Gran-Turismo | Photo: Lamborghini
General Motors Electric Vehicles Will Feature in Netflix Productions
News Anthony Lemonde - February 6, 2023
The automaker signed a deal to have its EVs appear in movies and shows "Where relevant". Productions in which GM vehicles will feature...
2024 BMW X5 and X6 Update Brings Facelift and Power
News Matt St-Pierre - February 8, 2023
The midsize SUVs gain more power. The PHEV X5 not only gets more power but its EV range increases too. Visual changes include...
The 2024 Volkswagen Atlas will show up this week
News Vincent Aubé - February 7, 2023
Don't expect an electrified drivetrain. This facelift should be based on the current generation. Will this new face be the last one of...
Bugatti Chiron Profilée Auctions for 9,7 Million Euro, Becomes Most Valuable New Car Ever
This is a one-of-one special edition of the Chiron which is the last Bugatti powered by a W16 engine ever. The €9,792,500 winning...
2024 Mazda CX-90 Priced from $45,900 in Canada
The new large-luxury SUV arrives this spring. It will be available in no fewer than nine trims. The all-new 2024 Mazda CX-90's main job...
Previous articleElon Musk Shares Video Of CyberTruck Window Not Breaking
Next articleNew 2021 Mercedes-Benz GLA Teased, Will Debut On December 11
The 2024 Toyota Grand Highlander introduced with more space and two hybrid engines
Charles Jolicoeur - February 8, 2023
The new 2024 Toyota Grand Highlander is a larger version with more cargo space than the current Highlander. The 2024 Toyota Grand Highlander will...
Subaru will Continue to Rely on Toyota for its Electric Vehicles
Anthony Lemonde - February 8, 2023
The only current electric vehicle in the Subaru lineup is the Solterra, an almost identical copy of the Toyota bZ4X. The automaker will...
Karma will Build B-ON Delivery Trucks in its California Factory
Karma and B-ON have teamed up to provide electric delivery vehicles for the North and South American markets. B-ON is a German manufacturer...
BMW, Hyundai, and Kia Popular in the World Car of the Year Finalist List
BMW has five entries out of 33 while Hyundai, Kia, and Genesis have eight entries. The winner of each category will be announced...
Mercedes-Benz unveils the 2024 eSprinter
Vincent Aubé - February 8, 2023
The battery pack has a capacity of (usable) 113 kWh. Two levels of power will be offered : 134 or 201 hp. The...
Matt St-Pierre - February 8, 2023
Motor Illustrated is all about cars. We want to inform consumers about the latest automotive news, and hopefully provide them with the info they need when buying a new or pre-owned car. We also tend to be pretty opinionated. MotorIllustrated.com is a property of NetMedia360 inc which is part of 360.Agency 355 Peel 9th floor Montreal, Quebec, H3C 2G9 Phone: 1 833 360-1955 360.Agency NetMedia360
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text:
Monkee Spectacular
One year ago, and twelve issues back, the Monkees had just begun to sizzle as the hottest group in show business. Monkee Spectacular made its debut on the newsstand to the delight of Monkee fans everywhere. We're happy to say the magazine is still going strong and will be around as long as the Monkees. And you can be sure that Davy, Peter, Micky and Mike are not about to forsake their fans for many years to come.
Currently the boys are shooting their feature film at Columbia studios. The movie will be released late this summer. In April the Monkees begin their Far Eastern tour. Monkee Spec will go right along with them to bring you all the excitement and color of this super-fab tour.
Concerning next year's TV show, don't forget to write to the Vice President in charge of programming, NBC-TV, Rockefeller Center, New York City, New York. Let him know how strongly you feel about the Monkees returning to TV in the fall! Do not write to the Monkees, or to Screen Gems. Your letters should go to NBC if you want the show back again.
Previous (March 1968)
Next (May 1968)
Magazine: Monkee Spectacular
Editor: Ralph Benner
Publisher: Laufer Publishing Co.


text:
Q: How to output large HTML chunk using JavaScript? hi all i am trying to output a large html chunk that holds opening and closing javascript tags. The javascript included is a jw player script and its mixed with javascript dynamic variable +url and php variable $Title. i tried document.write but the player didn't show up in the browser !instead it output part of the document.write statements! could you guys help me fix this issue ? Thanks in advance.
document.write("");
document.write("");
document.write("<meta http-equiv="Content-Type" content="text/html; charset=utf-8">");
document.write("");
document.write("
");

document.write("Title:<?php echo $Title;?> ");

document.write("
This text will be replaced
");
document.write("");
document.write("");

A: PHP written into the document with JavaScript will not run. PHP is evaluated before a document is received by the browser. JavaScript runs after.
As far as writing out a

This text will be replaced

Note  tag added.

A: Not sure what you're trying to do but document.getElementById("idtag").innerHTML = "Data"; will fill in the content between the tags of the element with the selected id.
Example:

redpajama_set_name: RedPajamaStackExchange


The Claddagh ring (Irish: fáinne Chladaigh) is a traditional Irish ring given which represents love, loyalty, and friendship (the hands represent friendship, the heart represents love, and the crown represents loyalty).
The design and customs associated with it originated in the Irish fishing village of Claddagh, located just outside the old city walls of Galway, now part of Galway City. The ring, as currently known, was first produced in the 17th century.
What is a Claddagh Ring?
There are very few people in Ireland who don't know what a claddagh ring is, but for those further overseas, it is a traditional Irish ring that was customary for lovers to give one another during the 19th and 20th century, although that wasn't how it originated. It is still popular today and often given as keepsakes, or for visitors to take home as souvenirs. The design is attritbuted to Richard Joyce, who was a slave in the West Indies when he was captured by mediterranean pirates. He was sold to a Moorish goldsmith who trained him, before setting up a jewellery business when he eventually got back to Galway in the 17th century.
The ring was adopted by the people of Claddagh, a village just outside the walls of Galway city (now more of a suburb of the city) whose inhabitants were well known for their staunch resistance of British influence and passion for sticking to their own customs, language, and way of life. The village was largely self sufficient, with the men heading out every day on boats known as Galway Hookers to catch fish from Galway Bay. When they returned, the women would bring the catch across the water to the city to sell. The fishermen of the village wore the ring as a sort of identification; so if anyone was found fishing in their territory and wasn't wearing the ring, there would be a lot of trouble.
Eventually the ring made its way across the country to Dublin, where fashionistas fell in love with it and it became a must have accessory. Designs became more ornate with added jewels, but the basic elements stayed the same; a heart held by two hands with a crown on top; because of this particular design certain symbolism that the claddagh ring is now famous for became established. The heart was attributed to love, the hands to friendship, and the crown to loyalty. The ring became a popular gift for friends and lovers to give each other, and with increased emigration spread across the water to Britain, the United States, Canada and even some further flung places. Claddagh rings became family heirlooms or were even used as wedding rings.
An etiquette of sorts emerged about how to wear the ring in regard to your relationship status, although how this started and where it came from isn't known. It all depends on which hand you wear the ring and in which direction. On the left hand, if the heart is pointing inwards towards your body, you are taken and it is a very serious relationship. On the right hand if the heart is pointing inwards, you have a special someone but you're not necessarily committed to them; the ring was usually worn this way if given by a friend. If the heart points outwards on a right hand, you're not in a relationship but you would like to be in one, and if it's pointing outwards on the left hand, you're definitely on the lookout for a spouse.
Walt Disney and his wife apparently purchased a pair of claddagh rings during a trip to Ireland, and wore them often. Walt's statue in Disneyland even has the claddagh ring included on his outstretched hand, but the heart is pointing outwards. Either someone made a grave mistake, or Walt didn't know how to wear his ring properly.


Essential oils are used for aromatherapy, topical application, and household use. This kit has the 8 vials of the most popular essential oils.
Each vial contains 100% pure, concentrated oils derived from all-natural plant extracts. Combined with superior distillation and bottling process each drop will refresh the air and reinvigorate your mind and body.
For aromatherapy, just add a few drops to a diffuser.
- ZEN made from Santalum: Our Zen essential oil blend is specially formulated to give you energy while keeping you calm and grounded.
- CLEANSE made from Lavendula spica, Metha x Piperita: Ability to remove toxins accumulated over time due to stress, environmental impact such as air & water pollution, pesticides and chemicals from cosmetics, foods, cookware, and fabrics.
- OM made from Boswellia carterii: To reduce anxiety. Put a couple of drops in or around your navel then spread it on the soles of your feet and on your clavicle and feel the Ohm.
- RELIEF made from Citrus Bergamia: Also known as the headache soother. Apply behind ears, on wrists, temples, the back of the neck & on the bottom of the feet.
- SOLACE made from Cedrus deodara: Actually help relieve cramping, nausea, hot flashes and moodiness, its aroma is floral and fruity in the most lovely way.
- BLISS made from Eucalyptus globulus: The essential oils in this must-have blend were meticulously chosen to energize you.
- SERENE made from Lavendula spica, Chamomile, Marjoram: Helps calm nerves, turns off overthinking and relaxes the mind.
- CHI made from Citrus x Sinensis, Juniper & Tea Tree: With tea tree oil - melaleuca alternifolia - Make lice, dandruff & flakes disappear. Use a few drops of the oil to make tea tree shampoo, tea tree lotion, and soap. A natural fungal infection treatment and rejuvenating aromatherapy essential oil.
They will also just make the room smell lovely as an air freshener! Use a few drops of your favorite, or combine to create custom scents.
Need some relief? Dab a spot on your body part that needs attention, such as your temples. Mix with a carrier oil for acne treatment and skin ailments. Or mix these oils with water to create custom, natural cleaning recipes.
- Topical application; Relieve Pain, Skin Treatment, etc.


text:
Q: Cannot create class file in Android Studio As you can see in the image below, when I try to create a class file this notification appear, I was looking for a solution here in SO and I checked other questions but none of them solved my problem, What do I need to do to solve this?
I hope you can help me, thank you.

A: I faced similar problem, and was able to resolve it by installing a fresh copy of Android Studio.
(A bit) More detailed answer is here.

A: I resolve this problem by deleting header generated automatically in a Java file.
Go to :
Preference ==> Search for header (in the top left input) ==>File header (make it empty)
Hope I help you


The Boys' Varsity Volleyball Team traveled to Christopher Columbus High School to square off against the host school and #2 seed in the district semifinal. During the regular season the Raiders lost a heartbreaking 5 set match against the Explorers. However, this time the story was different. In another thrilling 5 set match, the Raiders came out victorious and advanced to the district final on Wednesday night. Ryan Peluso led the charge with 25 kills, 4 aces and 2 blocks. Will Drody contributed 12 kills and Thomas Murphy recorded 6 kills and 5 blocks. Logan Courey added 4 kills and 4 blocks while Nicolas Stone Perez dished out a team-high 44 assists and served 2 aces. Rylan Sadler had a solid defensive effort with 9 digs.
The Boys' Varsity Volleyball Team traveled to Cardinal Gibbons High School for their last regular season game and defeated the defending state champions by a score of 3-2. Ryan Peluso led the Raiders with 20 kills and 6 digs while Will Drody had 18 kills of his own. Nicolas Stone Perez had a team-high 45 assists and Ryan Sadler led the team in digs with 7. Thomas Murphy contributed 3 kills and 4 blocks and Logan Courey added 2 kills and 2 blocks. Magnus Kron had 3 service aces.
Ryan Peluso had a team-high 40 kills as the Boys' Varsity Volleyball Team defeated Belen Jesuit on Senior Night, 3-1. Will Drody contributed 18 kills, 3 blocks and 2 aces while Thomas Murphy added 4 kills and 5 blocks. Nicolas Stone Perez led the team in assists with 61 and Logan Courey had 2 kills and 2 blocks, including the game-winning block to propel the Raiders to victory. Seniors Max Bolzan and Thomas Murphy were honored after their last regular season home game.
The Boys' Varsity Volleyball Team finished in 2nd place at the 11th Annual Smoothie King Invitational hosted by Ransom Everglades School. The team lost to Dr. Phillips High School in the Gold Division championship game in a hard fought match in three sets: 15-25, 25-20, 13-15. Ryan Peluso led the Raiders with 15 kills, 4 service aces and 6 digs while Will Drody added 8 kills. Thomas Murphy contributed 3 blocks and 2 kills. Drody, Murphy and Peluso were selected to the All-Tournament Team.
The Boys' Varsity Volleyball Team traveled to Orlando this weekend for the Dr. Phillips Tournament. The team advanced to the Gold Bracket after victories over Lyman and Boone High School. The team finished 4th overall. Tournament statistic totals include: Ryan Peluso (48 kills, 12 digs and 5 service aces), Will Drody (35 kills and 8 blocks), Nicolas Stone Perez (89 assists, 11 digs, 5 kills and 5 blocks), Logan Courey (12 blocks and 2 kills), Thomas Murphy (5 blocks and 1 kill), Nick Namias (9 digs, 3 blocks and 1 service ace), Magnus Kron (12 digs and 2 service aces) and Ben Cobb (10 digs and 1 service ace).


When I came to Julianna I was working out and eating better then I ever had in my life. Despite my new habits, my weight kept going up and I could not figure out why. When I got my RMR tested with Julianna and discovered how much food I needed to eat each day made all the difference. I finally was able to stop the weight gain and turn it in the opposite direction. I always thought that just by eating healthy and working out you will lose weight. Apparently it is not always that simple. I found out that I had allergies that were causing negative reactions with certain types of foods. Julianna was able to give me a specific list of foods that were good for me and others I should avoid, which is amazing. This whole experience has taught me so much that I would never have been able to figure out on my own. Julianna is amazing and just a joy to work with!


text:
Q: R:CrossTable- How to print frequencies for total row and column observations with xtable? I produced a cross table with the CrossTable command from the gmodels package such as:
library(gmodels)
library(descr)
a<-CrossTable(mtcars$cyl,mtcars$gear,prop.chisq = FALSE)

If I create a table for LaTex using xtable, the frequencies for row totals (last column) and column totals (last row) are missing. However, the total counts are there.
I used:
print(xtable(a,digits=3))

to achieve this. How do I get these values in as well?
Thanks for your support.
EDIT:
Here an example of the output:

a

===========================================
          mtcars$gear
mtcars$cyl        3       4       5   Total
-------------------------------------------
4                 1       8       2      11
              0.091   0.727   0.182   **0.344**
              0.067   0.667   0.400
              0.031   0.250   0.062
-------------------------------------------
6                 2       4       1       7
              0.286   0.571   0.143   **0.219**
              0.133   0.333   0.200
              0.062   0.125   0.031
-------------------------------------------
8                12       0       2      14
              0.857   0.000   0.143   **0.438**
              0.800   0.000   0.400
              0.375   0.000   0.062
-------------------------------------------
Total            15      12       5      32
              **0.469**   **0.375**   **0.156**
===========================================

I highlighted the missing values after xtable conversion with *. But when I have it converted to Tex with print it looks like:
\begin{table}[ht]
\centering
\begin{tabular}{llrrrr}
  \hline
 & mtcars\$cyl & 3 & 4 & 5 & Total \\
  \hline
1 & 4 & 1 & 8 & 2 & 11 \\
  2 &  & 0.091 & 0.727 & 0.182 &  \\
 3 &  & 0.067 & 0.667 & 0.400 &  \\
 4 &  & 0.031 & 0.250 & 0.062 &  \\
 5 & 6 & 2 & 4 & 1 & 7 \\
 6 &  & 0.286 & 0.571 & 0.143 &  \\
 7 &  & 0.133 & 0.333 & 0.200 &  \\
 8 &  & 0.062 & 0.125 & 0.031 &  \\
 9 & 8 & 12 & 0 & 2 & 14 \\
  10 &  & 0.857 & 0.000 & 0.143 &  \\
  11 &  & 0.800 & 0.000 & 0.400 &  \\
  12 &  & 0.375 & 0.000 & 0.062 &  \\
  13 & Total & 15 & 12 & 5 & 32 \\
   \hline
\end{tabular}
\end{table}

A: The output of str(a) suggests that the numbers that are missing in the print(xtable(a)) output are actually not part of the a object. The frequencies for row and column totals are calculated within the print.CrossTable method. Too see this, run getS3method("print","CrossTable"). Roughly in the middle of the output, you will see
if (prop.r) {
    ...
    m[k, nc + 1] <- formatC(hdd * RS[i]/GT, digits = digits,
            format = "f", decimal.mark = outDec)
    ...
}

i is the current row in the contingency table, k is the current output row (there are several output rows for each row in the contingency table), nc is the number of columns in the contingency table, RS[i] is the rows total of the current row of the contingency table, GT is the overall total, hdd is either 1 or 100 depending on the format of the output.
This does not solve the problem, but at least it explains why you see a different result with print(xtable(a)) than with simple print(a).

A: This could be viewed as a bug in the xtable.Crosstable method, which is in the descr package
environment(getS3method("xtable", "CrossTable"))
<environment: namespace:descr>

So you might like to contact the package maintainer about this. Meanwhile here is a hack that avoids re-computing the proportions yourself.
First capture the printed output from xtable to a temporary file and read each line back in as a separate string:
file <- tempfile()
capture.output(print(xtable(a, digits=3), include.rownames = FALSE),
               file = file)
xout <- readLines(file)

I have assumed you don't want the rownames in your table. Do the same for the printed version of the CrossTable:
capture.output(CrossTable(mtcars$cyl,mtcars$gear,prop.chisq = FALSE),
           file = file)
out <- readLines(file)

Next extract the body of the printed CrossTable output and "LaTeXize" it
body <- out[11:29]
## replace whitespace by column delimiter
body <- gsub("  *", " & ", body)
## add end of line markup for table rows
body <- gsub("([^-])$", "\\1 \\\\\\\\", body)
## replace dashed lines with hline
body <- gsub("-+", "\\\\hline", body)
## replace $ with \$
body <- gsub("$", "\\$", body, fixed = TRUE)

Finally print the header and footer from the printed xtable output with the new body:
cat(xout[1:6], body, xout[22:24], sep = "\n")

Producing the output
% latex table generated in R 3.1.1 by xtable 1.7-4 package
% Tue Oct 14 12:17:07 2014
\begin{table}[ht]
\centering
\begin{tabular}{lrrrr}
  \hline
mtcars\$cyl & 3 & 4 & 5 & Total \\
\hline
4 & 1 & 8 & 2 & 11 \\
 & 0.091 & 0.727 & 0.182 & 0.344 \\
 & 0.067 & 0.667 & 0.400 &  \\
 & 0.031 & 0.250 & 0.062 &  \\
\hline
6 & 2 & 4 & 1 & 7 \\
 & 0.286 & 0.571 & 0.143 & 0.219 \\
 & 0.133 & 0.333 & 0.200 &  \\
 & 0.062 & 0.125 & 0.031 &  \\
\hline
8 & 12 & 0 & 2 & 14 \\
 & 0.857 & 0.000 & 0.143 & 0.438 \\
 & 0.800 & 0.000 & 0.400 &  \\
 & 0.375 & 0.000 & 0.062 &  \\
\hline
Total & 15 & 12 & 5 & 32 \\
 & 0.469 & 0.375 & 0.156 \\
\hline
\end{tabular}
\end{table}


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VMAs 2020: Keke Palmer on BTS, True Jackson VP, and More
"It's overdue, baby."
By P. Claire Dodson
Photo: Chance Davis
When Keke Palmer announced her MTV VMAs 2020 hosting gig by reprising her role on True Jackson, VP, the actor and musician says she was surprised and touched by the response from fans of the Nickelodeon show all these years later.
"It touches my heart," she tells Teen Vogue. "I've had people come up to me and be like, I go to FIDM because of True Jackson. For me, I was just a kid trying to live my dream and do this whole entertainment thing. I didn't realize that it impacted people as much as it impacted me, just in a different way. It was awesome."
Hopefully we'll see more of True Jackson when Keke takes over the VMAs this Sunday, August 30. The notorious annual ceremony is known for its outrageous fashion and memorable viral moments, though those may be in shorter supply this year. The Video Music Awards will be held in various locations around New York City's five boroughs, but with limited audiences. We likely won't get much inter-celebrity action, which is typically a highlight.
But that doesn't mean the lineup is any less stacked. The night's performers include Ariana Grande and Lady Gaga (performing their hit "Rain on Me"), Miley Cyrus, The Weeknd, CNCO, Doja Cat, Maluma, Black Eyed Peas, and DaBaby. Oh, and of course, BTS, the seven-member Korean group that just broke YouTube's 24-hour video views record with their new single "Dynamite." The group will be performing the song for the first time during the ceremony.
"Yes, I'm a fan," Keke says of BTS. "My brother is the biggest K-pop fan and BTS is his favorite group. Same for my older sister, so I know a lot about them. We're all just shook because it's iconic for a K-pop group to be hitting the VMAs stage, and it's overdue, baby."
The VMAs have added two all-new categories this year — "Best Music Video From Home" and "Best Quarantine Performance" — which recognize "the artists who have released new music during quarantine to keep fans entertained while safely sheltered-in-place at home," according to a press release. Leading the nominees are Lady Gaga and Ariana, who both have nine nominations apiece, including Video of the Year and Song of the Year.
While the Video Music Awards will look different than years past because of the coronavirus pandemic, Keke hopes it will still be a celebratory moment in the landscape of an extremely bad year.
"That's the energy I'm putting into this," she says. "Not avoiding what's going on, and being very present with that. But at the same time offering this space where we can enjoy each other and come together and realize we're more alike than we're not. Just as much as negative is going on, there's positive going on too, because look how we respond to each other and hear each other and are real with each other. Let's keep this attitude more because that's what's going to get us through all this other stuff."
The MTV VMAs 2020 will air on Sunday, August 30 at 8 p.m. ET on MTV as well as on BET, BET Her, CMT, Comedy Central, Logo, MTV2, Nick at Nite, Paramount Network, Pop, TV Land, VH1, and The CW. Check out the full list of nominees here.
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Keke Palmer Shared Her Unseen Disney Channel Pilot With Vanessa Hudgens
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VMAs 2020: Keke Palmer Hosts, Nominees Include Ariana Grande, BTS, Lady Gaga
Keywordsvmas 2020vmasmtvBTSkeke palmer


� President Bush is sending his top aide on national security affairs to Capitol Hill today to confront what has become a tough crowd on the Iraq war.
A majority of senators, including key Republicans, believe troops should start coming home within the next few months. A new House investigation concluded this week that the Iraqis have little control over an ailing security force. And House Republicans are calling to revive the independent Iraq Study Group to give the nation options.
Lugar, the senior Republican on the Senate Foreign Relations Committee, plans to meet today privately with Bush�s national security adviser, Stephen Hadley. Hadley requested the meeting after Lugar delivered a lengthy floor speech contending the president�s war strategy won�t work and that U.S. troops should start leaving.
Other GOP senators, including Sen. George Voinovich of Ohio and Sens. Norm Coleman of Minnesota and John Sununu of New Hampshire also say they want to see troops departing Iraq by early 2008. Sen. Susan Collins, R-Maine, said yesterday she is working with Sens. John Warner, R-Va., and Ben Nelson, D-Neb., on a new bipartisan policy for Iraq.
In another challenge to Bush�s Iraq policy, House Republicans urged the White House yesterday to revive the Iraq Study Group. A blue-ribbon panel chaired by Republican James Baker and Democrat Lee Hamilton, the group concluded last December that U.S. troops could leave by March 2008 if certain steps were taken.
Last week, the House voted 355-69 to appropriate a $1 million budget for another study, though the bill is unlikely to become law for a few weeks. Reps. Chris Shays, R-Conn., and Frank Wolf, R-Va., said they hope the administration will move ahead on its own to reconvene the group.
The Republican push comes as a new bipartisan investigation found that Iraqi ministries are incapable of �accounting for, supporting or fully controlling their forces in the field.� The report also confirmed that the U.S. is shifting its focus from trying to transfer control to Iraqi troops � because they are not ready � to trying to secure neighborhoods.
�Basically, the department can tell us how many people they�ve trained and how many weapons they�re given, but they can�t tell us enough about how well they can perform their missions or even plan them,� said Rep. Marty Meehan, D-Mass., the outgoing chairman of the House Armed Services oversight and investigations subcommittee.