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Zebrafish hearts can regenerate by replacing damaged tissue with new cardiomyocytes. Although the steps leading up to the proliferation of surviving cardiomyocytes have been extensively studied, little is known about the mechanisms that control proliferation and redifferentiation to a mature state. We found that the cardiac dyad, a structure that regulates calcium handling and excitation-contraction coupling, played a key role in the redifferentiation process. A component of the cardiac dyad called leucine-rich repeat–containing 10 (Lrrc10) acted as a negative regulator of proliferation, prevented cardiomegaly, and induced redifferentiation. We found that its function was conserved in mammalian cardiomyocytes. This study highlights the importance of the underlying mechanisms required for heart regeneration and their application to the generation of fully functional cardiomyocytes.

Conservation of species facing environmental change requires an understanding of interpopulation physiological variation. However, physiological data are often scarce and therefore pooled across populations and species, erasing potentially important variability between populations. Interpopulation variation in thermal physiology has been observed within the Salmonidae family, although it has not been associated with seasonally distinct migratory phenotypes (i.e., seasonal runs). To resolve whether thermal physiology is associated with life-history strategy, we acclimated four Sacramento River juvenile Chinook salmon ( Oncorhynchus tshawytscha) populations (Coleman fall-run, Feather River fall-run, Feather River spring-run, and Sacramento River winter-run) exhibiting different seasonal migratory phenotypes (fall-, spring-, and winter-run), at 11, 16, and 20 °C and assessed variation in growth rate, critical thermal maxima, and temperature-dependent metabolic traits. We identified population differences in the physiological parameters measured and found compelling evidence that the critically endangered and endemic Sacramento River winter-run Chinook population exhibits thermal physiology associated with its early-migration life-history strategy. Acclimation to warm temperatures limited the growth and metabolic capacity of winter-run Chinook salmon, highlighting the risk of future environmental warming to this endemic population.

The lack of a long-term in vitro culture method has severely restricted the study of Plasmodium vivax, in part because it limits genetic manipulation and reverse genetics. We used the recently optimized Plasmodium cynomolgi Berok in vitro culture model to investigate the putative P. vivax drug resistance marker MDR1 Y976F. Introduction of this mutation using clustered regularly interspaced short palindromic repeats–CRISPR-associated protein 9 (CRISPR-Cas9) increased sensitivity to mefloquine, but had no significant effect on sensitivity to chloroquine, amodiaquine, piperaquine, and artesunate. To our knowledge, this is the first reported use of CRISPR-Cas9 in P. cynomolgi, and the first reported integrative genetic manipulation of this species.

Extirpation of the Central Valley spring-run Chinook salmon ( Oncorhynchus tshawytscha) evolutionary significant unit (ESU) from the San Joaquin River is emblematic of salmonid declines across the western seaboard of the United States. Habitat restoration and fish reintroduction efforts are ongoing, but recent telemetry studies have revealed low outmigration survival of juveniles to the ocean. Previous investigations have focused on modeling survival relative to river discharge and geographic regions, but have largely overlooked the effects of habitat variability. To evaluate the link between environmental conditions and survival of juvenile spring-run Chinook salmon, we combined high spatial resolution habitat mapping approaches with acoustic telemetry along a 150 km section of the San Joaquin River during the spring of 2019. While overall outmigration survival was low (5%), our habitat-based classification scheme described variation in survival of acoustic-tagged smolts better than other candidate models based on geography or distance. Two regional mortality sinks were evident along the longitudinal profile of the river, revealing poor survival in areas that shared warmer temperatures but that diverged in chlorophyll a, fluorescent dissolved organic matter (fDOM), turbidity, and dissolved oxygen levels. These findings demonstrate the value of integrating river habitat classification frameworks to improve our understanding of survival dynamics of imperiled fish populations.

LAY SUMMARY Some U.S. student Veterans pursuing higher education are incurring debt despite benefits available through the GI Bill. This exploratory study examines credit card debt accumulated by U.S. student Veterans while pursuing higher education goals. The purpose of this article is to address whether the U.S. GI Bill provides enough revenue for some students and to alert policy makers to the potential need to realign GI Bill benefits to help more student Veterans.

Malaria is a major global health problem which predominantly afflicts developing countries. Although many antimalarial therapies are currently available, the protozoan parasite causing this disease, Plasmodium spp., continues to evade eradication efforts. One biological phenomenon hampering eradication efforts is the parasite's ability to arrest development, transform into a drug-insensitive form, and then resume growth post-therapy. Currently, the mechanisms by which the parasite enters arrested development, or dormancy, and later recrudesces or reactivates to continue development, are unknown and the malaria field lacks techniques to study these elusive mechanisms. Since Plasmodium spp. salvage purines for DNA synthesis, we hypothesised that alkyne-containing purine nucleosides could be used to develop a DNA synthesis marker which could be used to investigate mechanisms behind dormancy. Using copper-catalysed click chemistry methods, we observe incorporation of alkyne modified adenosine, inosine, and hypoxanthine in actively replicating asexual blood stages of Plasmodium falciparum and incorporation of modified adenosine in actively replicating liver stage schizonts of Plasmodium vivax. Notably, these modified purines were not incorporated in dormant liver stage hypnozoites, suggesting this marker could be used as a tool to differentiate replicating and non-replicating liver forms and, more broadly, as a tool for advancing our understanding of Plasmodium dormancy mechanisms.

After applying canonical correspondence analysis to metagenomics data with hugely different library sizes (site totals) it became evident that Canoco and the R-packages ade4 and vegan can yield (at least up to 2022) very different P-values in statistical tests of the relationship between taxonomic composition (species composition) and predictors (environmental variables and/or treatments). The reason is that vegan and Canoco up to version 5.12 apply residualized response permutation (but ignore the model intercept), whereas ade4 applies predictor permutation. Predictor permutation, when extended to residualized predictor permutation, is applicable in partial constrained ordination. This paper shows by simulation that residualized response permutation can yield a very inflated Type I error rate, if the abundance data are both overdispersed and highly variable in site total. In contrast, residualized predictor permutation controlled the type I error rate and had good power, also when the predictors were skewed or binary. After square-root or log transformation of the abundance data, the differences between the permutation methods became small. Residualized predictor permutation is recommended, particularly in testing trait–environment relationships using double constrained correspondence analysis, because this method also critically depends on the species totals, which are generally highly variable. It is implemented in Canoco 5.15 and the R-code of this paper.

Radical cure of Plasmodium vivax malaria must include elimination of quiescent "hypnozoite" forms in the liver; however, the only FDA-approved treatments are contraindicated in many vulnerable populations. To identify new drugs and drug targets, we screened the Repurposing, Focused Rescue, and Accelerated Medchem library against P. vivax liver stages and identified the DNA methyltransferase inhibitors hydralazine and cadralazine as active against hypnozoites. We then used bisulfite sequencing and immunostaining to identify cytosine modifications in the infectious stage (sporozoites) and liver stages, respectively. A subsequent screen of epigenetic inhibitors revealed hypnozoites are broadly sensitive to histone acetyltransferase and methyltransferase inhibitors, indicating that several epigenetic mechanisms are likely modulating hypnozoite persistence. Our data present an avenue for the discovery and development of improved radical cure antimalarials.

This study presents the performance of a novel solar-powered incubator which uses water as thermal mass. Energy absorbed by the water is transferred to the incubator chamber by a fan powered with a 12 V battery. To enable adequate control of chamber temperature and humidity, a ProNem Mini sensor and controller (ESM-3723) were incorporated into the design. The thermal mass was sized adequately to supply energy for night operations as well as periods of inclement weather. The incubator was tested with and without load. Field measurements were obtained using UT330A USB datalogger. Tests under no load conditions gave an average temperature range of 36 to 42.9°C and relative humidity of 53 % to 70.5 %. Incubation tests showed average chamber temperature and relative humidity ranges of 35 ℃ to 41 ℃ and 45 % to 61 %. Candling test gave percentage fertility and hatchability of 60% and 56%. A cost-benefit analysis gave a capital cost of 617.32 USD, incubation cost of 69 cents per chick and simple payback period of 15 months. The performance indices obtained make the proposed design a suitable architecture to build upon in order to accelerate the promotion of livelihood empowerment through poultry farming in Ghana.

The purpose of this study was to evaluate the physical and psychological effects of COVID-related elective surgery delays on young sports medicine patients.We conducted a cross-sectional study of patients (10-25 years old) who had elective sports medicine surgery delayed due to the COVID crisis. Electronic surveys were sent to patients and included the 12-item Short Form Health Survey (SF-12), which yields a physical component score (SF12-PCS) and a mental component score (SF12-MCS), the PROMIS Psychological Stress Experience survey (PROMIS-PSE), and self-designed questions about patient concerns regarding the COVID crisis and delayed surgery.Of the 194 eligible patients with delayed elective sports surgeries, 107 patients (55%) elected to participate (mean age 17.6 ± 3.09 years, 30% male). The mean surgical delay was 76 days (CI 57-98). Delayed surgery patients scored significantly lower than population norms on the SF12-PCS (mean 39.3, CI 37.0-41.7;Young sports medicine patients reported significant physical and emotional symptoms associated with COVID-related surgical delays. Patients were most concerned about delays resulting in missed sport seasons. Those who reported greater levels of concern with surgical delays reported more emotional symptoms and higher levels of psychological stress.

Sexual minority youth (SMY) are at increased risk for interpersonal violence victimization compared to heterosexual youth. The current study examined how self-reported victimization (i.e., bullying, sexual harassment and dating violence) among middle school youth varied as a function of sexual/romantic attraction as well as gender identity. Cross-sectional data were gathered from students at seven middle schools in New England (

The nucleus in many cell types is a stiff organelle, but fat-filled lipid droplets (FDs) in cytoplasm are seen to indent and displace the nucleus. FDs are phase-separated liquids with a poorly understood interfacial tension γ that determines how FDs interact with other organelles. Here, micron-sized FDs remain spherical as they indent peri-nuclear actomyosin and the nucleus, while causing local dilution of Lamin-B1 independent of Lamin-A,C and sometimes triggering nuclear rupture. Focal accumulation of the cytosolic DNA sensor cGAS at the rupture site is accompanied by sustained mislocalization of DNA repair factors to cytoplasm, increased DNA damage, and delayed cell cycle. Macrophages show FDs and engulfed rigid beads cause similar indentation dilution. Spherical shapes of small FDs indicate a high γ, which we measure for FDs mechanically isolated from fresh adipose tissue as ∼40 mN/m. This value is far higher than that of protein condensates, but typical of oils in water and sufficiently rigid to perturb cell structures including nuclei.

Stream restorations are increasingly critical for managing and recovering freshwater biodiversity in human-dominated landscapes. However, few studies have quantifiedhow rehabilitative actions promulgate through aquatic communities over decades. Here, a long-term dataset is analyzed for fish assemblage change, incorporating data pre- and post-restoration periods, and testing the extent to which native assemblage stability increased over time. In the late 1950s, a large capacity dam was installed on Putah Creek (Solano County, CA, USA) which altered the natural flow regime, channel structure, geomorphic processes, and overall ecological function. Notably, downstream flows were reduced (especially during summer months) resulting in an aquatic assemblage dominated by warmwater nonnative species, while endemic native species subsisted at low levels as subordinates. A court-mediated Accord was ratified in 2000 providing a more natural flow regime, specifically for native and anadromous fishes in the stream. Richness of nonnative species decreased at every site following the Accord, while richness of native species increased or stayed constant. At the three most upstream sites, native species richness increased over time and ultimately exceeded nonnative richness. Native assemblage recovery was strongest upriver, closer to flow releases and habitat restoration activities, and decreased longitudinally downstream. Rank-abundance curves through time revealed that while species evenness was low throughout the study, dominance shifted from nonnative to native species in the upstream sites coincident with rehabilitation efforts. Mean rank shifts decreased following flow rehabilitation; thus the assemblage became increasingly stable over time following flow rehabilitation. Putah Creek’s rehabilitation may represent a model for others interested in improving endemic freshwater communities in degraded ecosystems.

A cell's mechanical environment can have many effects, but whether it impacts a cell's DNA sequence has remained unclear. To investigate this, we developed a live-cell method to measure changes in chromosome numbers. We edited constitutive genes with GFP/RFP-tags on single alleles and discovered that cells that lose Chromosome-reporters (ChReporters) become non-fluorescent. We applied our new tools to confined mitosis and to inhibition of the putative tumor suppressor Myosin-II. We quantified compression of mitotic chromatin in vivo and demonstrated that similar compression in vitro resulted in cell death, but also rare and heritable ChReptorter loss. Myosin-II suppression rescued lethal multipolar divisions and maximized ChReporter loss in 3D-compression and 2D-confinement, but not in standard 2D-culture. ChReporter loss associated with chromosome mis-segregation, rather than just the number of divisions, and loss in vitro and in mice was selected against in subsequent 2D-cultures. Inhibition of the spindle assembly checkpoint (SAC) caused ChReporter loss in 2D, as expected, but not in 3D-compression, suggesting a SAC perturbation. Thus, confinement and myosin-II affect DNA sequence and mechano-evolution, and ChReporters enable diverse studies of viable genetic changes.

Chromosomal instability (CIN), a state in which cells undergo mitotic aberrations that generate chromosome copy number variations, generates aneuploidy and is thought to drive cancer evolution. Although associated with poor prognosis and reduced immune response, CIN generates aneuploidy-induced stresses that could be exploited for immunotherapies. Macrophages, particularly, have been understudied in the CIN context. Here, through MPS1 inhibition-induced CIN in poorly immunogenic B16F10 mouse melanoma, we find that CIN- afflicted cancer cells skew macrophages towards an anti-cancer phenotype while also pushing them away from a pro-cancer one. We confirm these findings via RNA-sequencing, protein expression, and short-term tumor studies. These results further translate toin vivoefficacy in suppressing tumor growth: Mice can survive challenges of CIN-afflicted tumors. Long-term survival, however, is dependent on CD47 expression and IgG opsonization. Mice bearing CIN- afflicted tumors with wild-type CD47 levels see prolonged survival compared to their chromosomally stable counterparts, but all succumb. Mice bearing CIN-afflicted CD47 knockout tumors, however, show 28% long-term survival. When CD47 knockout was further paired with IgG opsonization, survival rate increased to 97%. Successful rejection and clearance of CIN- afflicted tumors inducedde novoanti-cancer IgG antibodies that were multi-epitope and functionally promoted macrophage-mediated phagocytosis. Thesede novoIgG antibodies could also suppressin vitrotumoroid andin vivotumor growth in a CD47 knockout context. These results highlight an unexpected therapeutic benefit from CIN when paired with maximal macrophage anti-cancer activity: an anti-cancer vaccination-like antibody response that can lead to durable cures and further potentiate cell-mediated acquired immunity.

Supplementary Table S5. Characteristics of the patient groups extracted from Wichmann et al.(64) and used in Figure 4. Patients were selected on basis of a negative HPV test result as HPV will confound the prognostic associations. All patients were treated with multimodality therapy regimens with curative intent.

Supplementary figures S1-S4

Patients with advanced stage head and neck squamous cell carcinoma (HNSCC) are often treated with cisplatin-containing chemoradiation protocols. Although cisplatin is an effective radiation sensitizer, it causes severe toxicity and not all patients benefit from the combination treatment. HNSCCs expectedly not responding to cisplatin may better be treated with surgery and postoperative radiation or cetuximab and radiation, but biomarkers to personalize chemoradiotherapy are not available. We performed an unbiased genome-wide functional genetic screen in vitro to identify genes that influence the response to cisplatin in HNSCC cells. By siRNA-mediated knockdown, we identified the Fanconi anemia/BRCA pathway as the predominant pathway for cisplatin response in HNSCC cells. We also identified the involvement of the SHFM1 gene in the process of DNA cross-link repair. Furthermore, expression profiles based on these genes predict the prognosis of radiation- and chemoradiation-treated head and neck cancer patients. This genome-wide functional analysis designated the genes that are important in the response of HNSCC to cisplatin and may guide further biomarker validation. Cisplatin imaging as well as biomarkers that indicate the activity of the Fanconi anemia/BRCA pathway in the tumors are the prime candidates. Mol Cancer Ther; 16(3); 540–50. ©2016 AACR.

Radioprotection in Egr1 knockout mice was also observed at an earlier time point.

Figure legends for supplementary figures

Knocking down Egr1 in normal cell lines reduces degree of apoptotic morphology after irradiation.

Legends to supplementary figures S1-S4

Purpose: Non–small cell lung cancer (NSCLC) and glioblastoma multiforme (GBM) have poor median survival. NSCLC and GBM overexpress glucose regulated protein 78 (GRP78), which has a role in radioresistance and recurrence. In this study, we determined the effect of anti-GRP78 antibody and the combined effect of the anti-GRP78 antibody with ionizing radiation (XRT) on NSCLC and GBM cell lines both in vitro and in vivo.Experimental Design: NSCLC and GBM cancer cell lines were treated with anti-GRP78 antibodies and evaluated for proliferation, colony formation, cell death, and PI3K/Akt/mTOR signaling. The efficacy of anti-GRP78 antibodies on tumor growth in combination with XRT was determined in vivo in mouse xenograft models.Results: GBM and NSCLC cells treated with anti-GRP78 antibodies showed attenuated cell proliferation, colony formation, and enhanced apoptosis. GBM and NSCLC cells treated with anti-GRP78 antibodies also showed global suppression of PI3K/Akt/mTOR signaling. Combining antibody with XRT resulted in significant tumor growth delay in both NSCLC and GBM heterotopic tumor models.Conclusions: Antibodies targeting GRP78 exhibited antitumor activity and enhanced the efficacy of radiation in NSCLC and GBM both in vitro and in vivo. GRP78 is a promising novel target, and anti-GRP78 antibodies could be used as an effective cancer therapy alone or in combination with XRT. Clin Cancer Res; 23(10); 2556–64. ©2016 AACR.

Supplementary Table from Targeting a Radiosensitizing Antibody–Drug Conjugate to a Radiation-Inducible Antigen

Supplementary figure legend

Purpose:We recently discovered that anti-TIP1 antibody activates endocytosis in cancer cells, which facilitates retention of antibody and dissociation of a conjugated drug. To improve the pharmacokinetics and cancer specificity of radiosensitizing drugs, we utilized antibody–drug conjugates (ADCs) that bind specifically to radiation-inducible antigen, TIP1, on non–small cell lung cancer (NSCLC). This approach exploits the long circulation time of antibodies to deliver a radiosensitizing drug to cancer each day during radiotherapy.Experimental Design:Antibodies to TIP1 were prioritized based on affinity, cancer-specific binding, and internalization. The lead antibody, 7H5, was conjugated with a cytotoxic drug MMAE because of its ability to radiosensitize cancer. Cytotoxicity, colony formation, and tumor growth studies were performed with 7H5-VcMMAE in combination with radiation.Results:7H5 showed a high affinity to recombinant TIP1 protein and radiation-inducible TIP1 on the cancer cell surface. 7H5 undergoes endocytosis in NSCLC cells in vitro. We obtained an average drug-to-antibody ratio (DAR) of 4.25 for 7H5-VcMMAE. A 70% reduction in viable cells was observed following 7H5-VcMMAE treatment compared with 7H5 alone in both A549 and H1299 cells. 7H5-VcMMAE sensitized NSCLC cells to radiation, thereby significantly decreasing the surviving fraction. The ADC combined with radiation showed a prolonged delay in tumor growth and improved survival in A549 and H1299 tumor models.Conclusions:Targeting radiation-inducible TIP1 with a radiosensitizing ADC is a promising strategy to enhance the therapeutic efficacy of NSCLC. This novel approach of targeting with ADCs to radiation-inducible antigens will lead to clinical trials in lung cancer patients treated with radiotherapy.

Treatment with antibody against GRP78 does not reduce proliferation and colony formation in normal cell lines.

Supplementary Figure from Targeting a Radiosensitizing Antibody–Drug Conjugate to a Radiation-Inducible Antigen

Supplementary Database 1 from Lithium Treatment Prevents Neurocognitive Deficit Resulting from Cranial Irradiation

Curative cancer treatment regimens often require cranial irradiation, resulting in lifelong neurocognitive deficiency in cancer survivors. This deficiency is in part related to radiation-induced apoptosis and decreased neurogenesis in the subgranular zone of the hippocampus. We show that lithium treatment protects irradiated hippocampal neurons from apoptosis and improves cognitive performance of irradiated mice. The molecular mechanism of this effect is mediated through multiple pathways, including Akt/glycogen synthase kinase-3β (GSK-3β) and Bcl-2/Bax. Lithium treatment of the cultured mouse hippocampal neurons HT-22 induced activation of Akt (1.5-fold), inhibition of GSK-3β (2.2-fold), and an increase in Bcl-2 protein expression (2-fold). These effects were sustained when cells were treated with lithium in combination with ionizing radiation. In addition, this combined treatment led to decreased expression (40%) of the apoptotic protein Bax. The additional genes regulated by lithium were identified by microarray, such as decorin and Birc1f. In summary, we propose lithium treatment as a novel therapy for prevention of deleterious neurocognitive consequences of cranial irradiation. (Cancer Res 2006; 66(23): 11179-86)

Supplementary Methods, Figures 1-3 from Targeted Nanoparticles That Deliver a Sustained, Specific Release of Paclitaxel to Irradiated Tumors

To capitalize on the response of tumor cells to XRT, we developed a controlled-release nanoparticle drug delivery system using a targeting peptide that recognizes a radiation-induced cell surface receptor. Phage display biopanning identified Gly-Ile-Arg-Leu-Arg-Gly (GIRLRG) as a peptide that selectively recognizes tumors responding to XRT. Membrane protein extracts of irradiated glioma cells identified glucose-regulated protein GRP78 as the receptor target for GIRLRG. Antibodies to GRP78 blocked the binding of GIRLRG in vitro and in vivo. Conjugation of GIRLRG to a sustained-release nanoparticle drug delivery system yielded increased paclitaxel concentration and apoptosis in irradiated breast carcinomas for up to 3 weeks. Compared with controls, a single administration of the GIRLRG-targeted nanoparticle drug delivery system to irradiated tumors delayed the in vivo tumor tripling time by 55 days (P = 0.0001) in MDA-MB-231 and 12 days in GL261 (P < 0.005). This targeting agent combines a novel recombinant peptide with a paclitaxel-encapsulating nanoparticle that specifically targets irradiated tumors, increasing apoptosis and tumor growth delay in a manner superior to known chemotherapy approaches. Cancer Res; 70(11); 4550–9. ©2010 AACR.

Supplementary Figures 1-3 from Lithium Treatment Prevents Neurocognitive Deficit Resulting from Cranial Irradiation