Cytosolic activation of cell death and stem rust resistance by cereal MLA-family CC-NLR proteins

Plants possess intracellular immune receptors designated “nucleotidebinding domain and leucine-rich repeat” (NLR) proteins that translate pathogen-specific recognition into disease-resistance signaling. The wheat immune receptors Sr33 and Sr50 belong to the class of coiled-coil (CC) NLRs. They confer resistance against a broad spectrum of field isolates of Puccinia graminis f. sp. tritici, including the Ug99 lineage, and are homologs of the barley powdery mildewresistance proteinMLA10. Here, we showthat, similarly to MLA10, the Sr33 and Sr50 CC domains are sufficient to induce cell death in Nicotiana benthamiana. Autoactive CC domains and full-length Sr33 and Sr50 proteins self-associate in planta. In contrast, truncated CC domains equivalent in size to an MLA10 fragment for which a crystal structure was previously determined fail to induce cell death and do not self-associate. Mutations in the truncated region also abolish self-association and cell-death signaling. Analysis of Sr33 and Sr50 CC domains fused to YFP and either nuclear localization or nuclear export signals in N. benthamiana showed that cell-death induction occurs in the cytosol. In stable transgenic wheat plants, full-length Sr33 proteins targeted to the cytosol provided rust resistance, whereas nuclear-targeted Sr33 was not functional. These data are consistent with CC-mediated induction of both cell-death signaling and stem rust resistance in the cytosolic compartment, whereas previous research had suggested that MLA10-mediated cell-death and disease resistance signaling occur independently, in the cytosol and nucleus, respectively

Fast-evolving noncoding sequences in the human genome

BACKGROUND: Gene regulation is considered one of the driving forces of evolution. Although protein-coding DNA sequences and RNA genes have been subject to recent evolutionary events in the human lineage, it has been hypothesized that the large phenotypic divergence between humans and chimpanzees has been driven mainly by changes in gene regulation rather than altered protein-coding gene sequences. Comparative analysis of vertebrate genomes has revealed an abundance of evolutionarily conserved but noncoding sequences. These conserved noncoding (CNC) sequences may well harbor critical regulatory variants that have driven recent human evolution. RESULTS: Here we identify 1,356 CNC sequences that appear to have undergone dramatic human-specific changes in selective pressures, at least 15% of which have substitution rates significantly above that expected under neutrality. The 1,356 'accelerated CNC' (ANC) sequences are enriched in recent segmental duplications, suggesting a recent change in selective constraint following duplication. In addition, single nucleotide polymorphisms within ANC sequences have a significant excess of high frequency derived alleles and high F(ST)values relative to controls, indicating that acceleration and positive selection are recent in human populations. Finally, a significant number of single nucleotide polymorphisms within ANC sequences are associated with changes in gene expression. The probability of variation in an ANC sequence being associated with a gene expression phenotype is fivefold higher than variation in a control CNC sequence. CONCLUSION: Our analysis suggests that ANC sequences have until very recently played a role in human evolution, potentially through lineage-specific changes in gene regulation

Reducing uncertainty with flood frequency analysis: the contribution of palaeoflood and historical flood information

Using a combination of stream gauge, historical, and paleoflood records to extend extreme flood records has proven to be useful in improving flood frequency analysis (FFA). The approach has typically been applied in localities with long historical records and/or suitable river settings for paleoflood reconstruction from slack‐water deposits (SWDs). However, many regions around the world have neither extensive historical information nor bedrock gorges suitable for SWDs preservation and paleoflood reconstruction. This study from subtropical Australia demonstrates that confined, semialluvial channels such as macrochannels provide relatively stable boundaries over the 1000–2000 year time period and the preserved SWDs enabled paleoflood reconstruction and their incorporation into FFA. FFA for three sites in subtropical Australia with the integration of historical and paleoflood data using Bayesian Inference methods showed a significant reduction in uncertainty associated with the estimated discharge of a flood quantile. Uncertainty associated with estimated discharge for the 1% Annual Exceedance Probability (AEP) flood is reduced by more than 50%. In addition, sensitivity analysis of possible within‐channel boundary changes shows that FFA is not significantly affected by any associated changes in channel capacity. Therefore, a greater range of channel types may be used for reliable paleoflood reconstruction by evaluating the stability of inset alluvial units, thereby increasing the quantity of temporal data available for FFA. The reduction in uncertainty, particularly in the prediction of the ≤1% AEP design flood, will improve flood risk planning and management in regions with limited temporal flood data

Characterization of highly stable liposomal and immunoliposomal formulations of vincristine and vinblastine

Liposome and immunoliposome formulations of two vinca alkaloids, vincristine and vinblastine, were prepared using intraliposomal triethylammonium sucroseoctasulfate and examined for their ability to stabilize the drug for targeted drug delivery in vivo. The pharmacokinetics of both the encapsulated drug (vincristine or vinblastine) and liposomal carrier were examined in Sprague Dawley rats, and the in vivo drug release rates determined. Anti-HER2 immunoliposomal vincristine was prepared from a human anti-HER2/neu scFv and studied for targeted cytotoxic activity in cell culture, and antitumor efficacy in vivo. Nanoliposome formulations of vincristine and vinblastine demonstrated similar pharmacokinetic profiles for the liposomal carrier, but increased clearance for liposome encapsulated vinblastine (t 1/2 = 9.7 h) relative to vincristine (t 1/2 = 18.5 h). Immunoliposome formulations of vincristine targeted to HER2 using an anti-HER2 scFv antibody fragment displayed a marked enhancement in cytotoxicity when compared to non-targeted liposomal vincristine control; 63- or 253-fold for BT474 and SKBR3 breast cancer cells, respectively. Target-specific activity was also demonstrated in HER2-overexpressing human tumor xenografts, where the HER2-targeted formulation was significantly more efficacious than either free vincristine or non-targeted liposomal vincristine. These results demonstrate that active targeting of solid tumors with liposomal formulations of vincristine is possible when the resulting immunoliposomes are sufficiently stabilized

Finishing the euchromatic sequence of the human genome

The sequence of the human genome encodes the genetic instructions for human physiology, as well as rich information about human evolution. In 2001, the International Human Genome Sequencing Consortium reported a draft sequence of the euchromatic portion of the human genome. Since then, the international collaboration has worked to convert this draft into a genome sequence with high accuracy and nearly complete coverage. Here, we report the result of this finishing process. The current genome sequence (Build 35) contains 2.85 billion nucleotides interrupted by only 341 gaps. It covers ∼99% of the euchromatic genome and is accurate to an error rate of ∼1 event per 100,000 bases. Many of the remaining euchromatic gaps are associated with segmental duplications and will require focused work with new methods. The near-complete sequence, the first for a vertebrate, greatly improves the precision of biological analyses of the human genome including studies of gene number, birth and death. Notably, the human enome seems to encode only 20,000-25,000 protein-coding genes. The genome sequence reported here should serve as a firm foundation for biomedical research in the decades ahead

Effect of angiotensin-converting enzyme inhibitor and angiotensin receptor blocker initiation on organ support-free days in patients hospitalized with COVID-19

IMPORTANCE Overactivation of the renin-angiotensin system (RAS) may contribute to poor clinical outcomes in patients with COVID-19. Objective To determine whether angiotensin-converting enzyme (ACE) inhibitor or angiotensin receptor blocker (ARB) initiation improves outcomes in patients hospitalized for COVID-19. DESIGN, SETTING, AND PARTICIPANTS In an ongoing, adaptive platform randomized clinical trial, 721 critically ill and 58 non–critically ill hospitalized adults were randomized to receive an RAS inhibitor or control between March 16, 2021, and February 25, 2022, at 69 sites in 7 countries (final follow-up on June 1, 2022). INTERVENTIONS Patients were randomized to receive open-label initiation of an ACE inhibitor (n = 257), ARB (n = 248), ARB in combination with DMX-200 (a chemokine receptor-2 inhibitor; n = 10), or no RAS inhibitor (control; n = 264) for up to 10 days. MAIN OUTCOMES AND MEASURES The primary outcome was organ support–free days, a composite of hospital survival and days alive without cardiovascular or respiratory organ support through 21 days. The primary analysis was a bayesian cumulative logistic model. Odds ratios (ORs) greater than 1 represent improved outcomes. RESULTS On February 25, 2022, enrollment was discontinued due to safety concerns. Among 679 critically ill patients with available primary outcome data, the median age was 56 years and 239 participants (35.2%) were women. Median (IQR) organ support–free days among critically ill patients was 10 (–1 to 16) in the ACE inhibitor group (n = 231), 8 (–1 to 17) in the ARB group (n = 217), and 12 (0 to 17) in the control group (n = 231) (median adjusted odds ratios of 0.77 [95% bayesian credible interval, 0.58-1.06] for improvement for ACE inhibitor and 0.76 [95% credible interval, 0.56-1.05] for ARB compared with control). The posterior probabilities that ACE inhibitors and ARBs worsened organ support–free days compared with control were 94.9% and 95.4%, respectively. Hospital survival occurred in 166 of 231 critically ill participants (71.9%) in the ACE inhibitor group, 152 of 217 (70.0%) in the ARB group, and 182 of 231 (78.8%) in the control group (posterior probabilities that ACE inhibitor and ARB worsened hospital survival compared with control were 95.3% and 98.1%, respectively). CONCLUSIONS AND RELEVANCE In this trial, among critically ill adults with COVID-19, initiation of an ACE inhibitor or ARB did not improve, and likely worsened, clinical outcomes. TRIAL REGISTRATION ClinicalTrials.gov Identifier: NCT0273570

A Great Promise and a Great Threat : Milwaukee Children in the Great Depression

I am a high school boy ... my father earns a salary that keeps us living, but does not allow for extras, a boy named Art explained in 1937. He wrote the Milwaukee Journal protesting the new fees the city had placed on recreational facilities to make up for budget cuts. The new tax slammed on tennis courts and swimming pools takes away from me the two sports I enjoy most .... Last summer I was frequently (almost daily) in the habit of swimming in the afternoon and playing tennis in the evening, or vise versa. Art argued that the city recreation facilities were established to keep people out of mischief and off the streets, but the new fees would prevent him and other children from using the pools and tennis courts. Art wondered what he would do with his extra free time. This summer I will probably have to lounge about street corners, pool halls or stroll through the parks trying to \u27pick up\u27 girlfriends. It does not sound pretty, but forced idleness is not pleasant. \u27..

A great promise and a great threat : Milwaukee children in the Great Depression

In the first decades of the twentieth century child advocates defined the ideal childhood. They argued that each child needed a stable home life, a quality education, and to be protected from the exploitation of work. Adults additionally, asserted that children needed the proper guidance to help them transition from childhood to maturity. Between 1900 and 1929, adults created laws, policies, and institutions to ensure boys and girls had proper childhood. The Great Depression robbed many children of this model life, causing reformers grave alarm. They turned to the federal government for assistance and used New Deal programs to aid the poorest children, end most forms of child labor, provide many jobless youth employment, and improve the education system. Adults saw the New Deal as a vital component in returning the model childhood to the nation\u27s youngest citizens. While these efforts helped they were not enough. By the mid-1930s idle children were committing crimes in record numbers, forcing juvenile justice officials to devise a low-cost crime-fighting strategy. They passed resolutions protecting girls and boys from bad influences such as salacious movies. They also used federal money to create the Toy Loan Project, which lent poor children toys to keep them out of trouble. The Great Depression also turned many children into activists. Needy children worked to change school culture so it was more accommodating to underprivileged students and sculpted the New Deal to meet their needs. Middle class youngsters contributed to charities that aided the poor and led the peace movement. As the Great Depression came to a close, adults believed that children had become radicals and criminals. In response, adults redefined citizenship so it emphasized American ideals and loyalty. While educators taught this new concept of citizenship, government officials began preparing youth to defend the nation. By the late 1930s, as the threat of fascism grew, children turned away from the anti-war movement to embrace the new patriotism. The approaching war also revitalized the economy and reformers saw the ideal childhood return to many children

Identification of Intracellular Calcium Oxalate Crystals in Chamelaucium Uncinatum (Myrtaceae)

Intracellular inclusions in the pedicel and calyx-tube tissues of Chamelaucium uncinatum Schauer (Myrtaceae) flowers are irregular in shape. They were shown, by polarised light and scanning electron microscopy, to be birefringent 8.9–29.5 μm druse (i.e. aggregate) crystals. Energy-dispersive X-ray spectroscopy showed that these crystals were predominantly composed of calcium. Histochemical and acid-solubility tests indicated that the crystals were calcium oxalate. Raman microprobe spectroscopy was used to confirm this chemical identity. The calcium oxalate crystals were located in xylem-vessel lumens and also in parenchyma cells adjacent to vascular tissues. Thus, the crystals may function to regulate soluble calcium concentrations in C. uncinatum tissues near sites where calcium is unloaded from the xylem