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Increased adoption of crops with stacked traits conferring glyphosate and dicamba resistance, and recent confirmation of kochia [Bassia scoparia (L.) A.J. Scott] biotypes resistant to these herbicides in Alberta and Saskatchewan, warrant surveillance of herbicide-resistant kochia in Manitoba. A randomized–stratified survey of 315 sites in Manitoba was conducted in the fall of 2018. Overall, 58% of the kochia populations tested were glyphosate-resistant, while 1% were dicamba-resistant. This survey documents rapid increase in glyphosate-resistant kochia over a five-year time frame, and also confirms the first cases of kochia in Manitoba with dicamba resistance alone and in combination with glyphosate resistance.

Recent confirmation of dicamba-resistant kochia [Bassia scoparia (L.) A.J. Scott] in Alberta warrants investigation of resistance to other commonly used synthetic auxin herbicides like fluroxypyr. A randomized-stratified survey of 305 sites in Alberta was conducted in 2017 to determine the status of fluroxypyr-resistant kochia. Overall, 13% of the kochia populations were fluroxypyr-resistant. Only 4% of the populations were both fluroxypyr- and dicamba-resistant, indicating that different mechanisms may confer resistance to these herbicides. When combined with estimates of dicamba resistance, about 28% of the kochia populations sampled in Alberta in 2017 were resistant to at least one synthetic auxin herbicide.

Recent confirmations of glyphosate-resistant Russian thistle (Salsola tragus L.) in Montana, Washington, and Oregon, warrant greater surveillance of herbicide-resistant Russian thistle in western Canada. A randomized-stratified survey of 315 sites in Manitoba was conducted in 2018 to determine the incidence of herbicide resistance in Russian thistle and other weeds sampled post-harvest. Russian thistle populations were collected from 14 of the 315 sites surveyed. None of these populations exhibited resistance to acetolactate synthase inhibitors (tribenuron/thifensulfuron), synthetic auxins (2,4-D ester or fluroxypyr), or glyphosate. This Manitoba survey of herbicide-resistant Russian thistle serves as a baseline for future surveillance efforts.

An assessment is made of published data on concentrations of four toxic trace elements arsenic (As), cadmium (Cd), lead (Pb), mercury (Hg), and methyl mercury (MeHg) in seafood species from Algerian coastal waters. The data relate mainly to fish and mollusks, with Pb and Cd being the most studied elements. In contrast, no studies were found for As and MeHg in mollusks and crustaceans, and none for total Hg in crustaceans. Data analysis has shown that the element concentrations depend on the element analyzed, in particular its chemical form, and they vary with the species studied and the area where the organisms were sampled. As concentrations in muscle of sardine (Sardina pilchardus) from the eastern zone (Bejaia) (2.98 mg kg−1 wet weight), Pb in muscle of tuna (Euthynnus alletteratus) (0.49 mg kg−1 wet weight) from the western region (Mostaganem) and in muscle tissue of the mussel (Mytilus galloprovincialis) (10.67 mg kg−1 dry weight) from Oran all exceed the threshold risk limits set by national and international regulations. Given these findings, to guarantee seafood safety for the Algerian consumer, it is recommended to carry out monitoring of these trace elements in seafood species caught along the Algerian coast, particularly the most consumed fish, i.e., sardines, and predators such as swordfish and tuna which can accumulate high concentrations of toxic metals, particularly Hg. Obtaining a regional assessment based on similar analyses from Tunisia and Morocco was severely hampered since few similar species or taxonomic groups were analyzed in all three countries.

Patients with familial arrhythmogenic cardiomyopathy typically present with ventricular arrhythmias or progressive heart failure. This paper characterizes a rare presentation of an underlying genetic cardiomyopathy with clinical manifestations mimicking an acute myocardial infarction in 2 siblings, each with the same mutation in the desmoplakin (DSP) gene. (Level of Difficulty: Advanced.), Central Illustration

Chronic lung disease has been attributed to stem cell aging and/or exhaustion. We investigated these mechanisms using mouse and human tracheobronchial tissue‐specific stem cells (TSC). In mouse, chromatin labeling and flow cytometry demonstrated that naphthalene (NA) injury activated a subset of TSC. These activated TSC continued to proliferate after the epithelium was repaired and a clone study demonstrated that ~96% of activated TSC underwent terminal differentiation. Despite TSC attrition, epithelial repair after a second NA injury was normal. The second injury accelerated proliferation of previously activated TSC and a nucleotide‐label retention study indicated that the second injury recruited TSC that were quiescent during the first injury. These mouse studies indicate that (a) injury causes selective activation of the TSC pool; (b) activated TSC are predisposed to further proliferation; and (c) the activated state leads to terminal differentiation. In human TSC, repeated proliferation also led to terminal differentiation and depleted the TSC pool. A clone study identified long‐ and short‐lived TSC and showed that short‐lived TSC clones had significantly shorter telomeres than their long‐lived counterparts. The TSC pool was significantly depleted in dyskeratosis congenita donors, who harbor mutations in telomere biology genes. The remaining TSC had short telomeres and short lifespans. Collectively, the mouse and human studies support a model in which epithelial injury increases the biological age of the responding TSC. When applied to chronic lung disease, this model suggests that repeated injury accelerates the biological aging process resulting in abnormal repair and disease initiation., Biological aging of tracheobronchial tissue specific stem cells (TSC) and their trophic unit, the pseudostratified conducting airway epithelium. Each injury activates a subset of TSC which proliferate and undergo terminal differentiation. Thus, each injury cycle depletes the TSC pool and many injuries compromise epithelial regeneration. Over time, biological age exceeds chronological age and increases the risk of chronic lung disease.

Major Histocompatibility Complex Class II (MHCII) is dynamically expressed on intestinal epithelial cells (IECs) throughout the intestine, but its regulation remains poorly understood. We observed that spontaneous upregulation of IEC MHCII in locally-bred Rag1−/− mice correlated with serum Interleukin (IL)-18, was transferrable via cohousing to commercially-bred immunodeficient mice and could be inhibited by both IL-12 and IL-18 blockade. Overproduction of intestinal IL-18 due to an activating Nlrc4 mutation upregulated IEC MHCII via classical inflammasome machinery independently of immunodeficiency or dysbiosis. Immunodeficient dysbiosis increased Il18 transcription, which synergized with NLRC4 inflammasome activity to drive elevations in serum IL-18. This IL-18-MHCII axis was confirmed in several other models of intestinal and systemic inflammation. Elevated IL-18 reliably preceded MHCII upregulation, suggesting an indirect effect on IECs, and mice with IL-18 overproduction showed activation or expansion of type 1 lymphocytes. Interferon Gamma (IFNg) was uniquely able to upregulate IEC MHCII in enteroid cultures and was required for MHCII upregulation in several in vivo systems. Thus, we have linked intestinal dysbiosis, systemic inflammation, and inflammasome activity to IEC MHCII upregulation via an intestinal IL-18-IFNg axis. Understanding this process may be crucial for determining the contribution of IEC MHCII to intestinal homeostasis, host defense, and tolerance., Graphical Abstract

Supplementary Data Contina, A., A. K. Pierce, S. W. Yanco, E. S. Bridge, J. F. Kelly, and M. B. Wunder. 2022. Application of stable isotopes to study movement ecology and diet variation in a migratory songbird. Journal of Field Ornithology. Accepted. https://journal.afonet.org/volXX/issYY/artZZ/ ABSTRACT Whether and how migratory organisms exhibit interindividual behavioral and/or physiological variation across movement strategies remains an open question. The Dark-eyed Junco (Junco hyemalis) is a migratory songbird known for its intraspecies variation displayed in relation to morphology, song repertoires, and migration. Thus, studies focusing on juncos can reveal how migratory strategy may covary with other individual-scale factors and, therefore, identify the selective forces driving intraspecies variations throughout its distribution. We used Dark-eyed Junco hydrogen stable isotope feather values (δ²H) and implemented a Bayesian framework to infer the breeding and molting origin of migratory juncos captured on their winter grounds in Oklahoma, USA. We modeled the distribution of feather hydrogen stable isotope values as a function of five morphological variables including body mass and fat deposition measured in Oklahoma during the winter. We then investigated the trade-off between longer and more energetically costly migration strategies, in relation to diet preferences through carbon (δ13C) and nitrogen (δ15N) stable isotope analysis, from feather values. Dark-eyed Juncos wintering in south central U.S. likely originate from multiple breeding populations in northern U.S. and Canada. Body condition at the wintering ground (e.g., mass) had no effect on feather hydrogen stable isotope abundance. However, we found a positive correlation between nitrogen and hydrogen stable isotopes, suggesting that a trophic level shift toward insect consumption might occur in individuals migrating from southern latitudes. Increased insect-derived protein consumption might be explained by reduced fatty-acid reserves necessary to complete a shorter migratory journey. Key words: avian migration; hydrogen; linear models; posterior probabilities; stable isotope

Herbicide-resistant (HR) kochia is a growing problem in the Great Plains region of Canada and the United States. Resistance to up to four herbicide sites of action, including photosystem II inhibitors, acetolactate synthase inhibitors, synthetic auxins, and the 5-enolpyruvylshikimate-3-phosphate synthase inhibitor glyphosate have been reported in many areas of this region. Despite being present in the United States since 1993/1994, auxinic-HR kochia is a recent and growing phenomenon in Canada. This study was designed to characterize 1) the level of resistance and 2) patterns of cross-resistance to dicamba and fluroxypyr in 12 putative auxinic-HR kochia populations from western Canada. The incidence of dicamba-resistant individuals ranged among populations from 0% to 85%, while fluroxypyr-resistant individuals ranged from 0% to 45%. In whole-plant dose-response bioassays, the populations exhibited up to 6.5-fold resistance to dicamba and up to 51.5-fold resistance to fluroxypyr based on visible injury 28 d after application. Based on plant survival estimates, the populations exhibited up to 3.7-fold resistance to dicamba and up to 72.5-fold resistance to fluroxypyr. Multiple patterns of synthetic auxin resistance were observed, in which one population from Cypress County, Alberta, was resistant to dicamba but not fluroxypyr, whereas another from Rocky View County, Alberta, was resistant to fluroxypyr but not dicamba based on single-dose population screening and dose-response bioassays. These results suggest that multiple mechanisms may confer resistance to dicamba and/or fluroxypyr in Canadian kochia populations. Further research is warranted to determine these mechanisms. Farmers are urged to adopt proactive nonchemical weed management practices in an effort to preserve efficacy of the remaining herbicide options available for control of HR kochia.

The UAB Nathan Shock Center focuses on comparative energetics and aging. Energetics, as defined for this purpose, encompasses the causes, mechanisms, and consequences of the acquisition, storage, and use of metabolizable energy. Comparative energetics is the study of metabolic processes at multiple scales and across multiple species as it relates to health and aging. The link between energetics and aging is increasingly understood in terms of dysregulated mitochondrial function, altered metabolic signaling, and aberrant nutrient responsiveness with increasing age. The center offers world-class expertise in comprehensive, integrated energetic assessment and analysis from the level of the organelle to the organism and across species from the size of worms to rats as well as state-of-the-art data analytics. The range of services offered by our three research cores, (1) The Organismal Energetics Core, (2) Mitometabolism Core, and (3) Data Analytics Core, is described herein.

The khapra beetle, Trogoderma granarium, is an important quarantine pest of stored-products. While it is not established in the North America, frequent interceptions highlight the need for treatments to eradicate isolated populations if they occur. Methyl bromide has long been the standard fumigant used for this purpose; however, increasing restrictions on its use necessitate the development of new treatment options. Here we evaluate a treatment for khapra beetle using a combination of sulfuryl fluoride (SF) and propylene oxide (PPO). Experiments were conducted to determine the most tolerant stages and the effective rates for each compound. Combination treatments using both products were then evaluated at several temperatures to determine effective doses for quarantine level control. The egg stage was by far the most tolerant to SF, requiring concentration × time (CT) products roughly 10-fold greater than the next most tolerant stage, diapausing larvae. Diapausing larvae were significantly more tolerant to SF than non-diapausing larvae in a recently collected ‘field’ strain, but not in a more than 30-yr-old ‘laboratory’ culture. PPO treatments were shown to completely control the egg stage at CTs greater than 155 hr-mgL-1 at both 10 and 20°C. Resulting combination treatments with initial dose rates of 96 and 40 mgL-1 at 10°C and 80 and 40 mgL-1 at 20°C are proposed for SF and PPO, respectively. These combination treatments utilize SF’s efficacy against the larval stage along with PPO’s superior ovicidal properties to provide enhanced treatment efficacy over each product alone against all life stages of khapra beetle.

In grasslands worldwide, modified fire cycles are accelerating herbaceous species extinctions. Fire may avert population declines by increasing survival, reproduction, or both. Survival and growth after fires may be promoted by removal of competitors or biomass and increasing resource availability. Fire-stimulated reproduction may also contribute to population growth through bolstered recruitment. We quantified these influences of fire on population dynamics in Echinacea angustifolia, a perennial forb in North American tallgrass prairie. We first used four datasets, 7–21 years long, to estimate fire’s influences on survival, flowering, and recruitment. We then used matrix projection models to estimate growth rates across several burn frequencies in five populations, each with one to four burns over 15 years. Finally, we estimated the contribution of fire-induced changes in each vital rate to changes in population growth. Population growth rates generally increased with burning. The demographic process underpinning these increases depended on juvenile survival. In populations with high juvenile survival, fire-induced increases in seedling recruitment and juvenile survival enhanced population growth. However, in populations with low juvenile survival, small changes in adult survival drove growth rate changes. Regardless of burn frequencies, our models suggest populations are declining and that recruitment and juvenile survival critically influence population response to fire. However, crucially, increased seedling recruitment only increases population growth rates when enough new recruits reach reproductive maturity. The importance of recruitment and juvenile survival is especially relevant for small populations in fragmented habitats subject to mate-limiting Allee effects and inbreeding depression, which reduce recruitment and survival, respectively.

Constitutive MAPK activation silences genes required for iodide uptake and thyroid hormone biosynthesis in thyroid follicular cells. Accordingly, most BRAFV600E papillary thyroid cancers (PTC) are refractory to radioiodide (RAI) therapy. MAPK pathway inhibitors rescue thyroid-differentiated properties and RAI responsiveness in mice and patient subsets with BRAFV600E-mutant PTC. TGFB1 also impairs thyroid differentiation and has been proposed to mediate the effects of mutant BRAF. We generated a mouse model of BRAFV600E-PTC with thyroid-specific knockout of the Tgfbr1 gene to investigate the role of TGFB1 on thyroid-differentiated gene expression and RAI uptake in vivo. Despite appropriate loss of Tgfbr1, pSMAD levels remained high, indicating that ligands other than TGFB1 were engaging in this pathway. The activin ligand subunits Inhba and Inhbb were found to be overexpressed in BRAFV600E-mutant thyroid cancers. Treatment with follistatin, a potent inhibitor of activin, or vactosertib, which inhibits both TGFBR1 and the activin type I receptor ALK4, induced a profound inhibition of pSMAD in BRAFV600E-PTCs. Blocking SMAD signaling alone was insufficient to enhance iodide uptake in the setting of constitutive MAPK activation. However, combination treatment with either follistatin or vactosertib and the MEK inhibitor CKI increased 124I uptake compared to CKI alone. In summary, activin family ligands converge to induce pSMAD in Braf-mutant PTCs. Dedifferentiation of BRAFV600E-PTCs cannot be ascribed primarily to activation of SMAD. However, targeting TGFβ/activin-induced pSMAD augmented MAPK inhibitor effects on iodine incorporation into BRAF tumor cells, indicating that these two pathways exert interdependent effects on the differentiation state of thyroid cancer cells.

Transaminase elevations can occur from liver injury or in the setting of rhabdomyolysis. The goal of this study is to evaluate indices that could differentiate acetaminophen toxicity from muscle injury in the setting of transaminase elevations.A retrospective chart review of consecutive cases reported to our regional poison center. Patients with transaminase (AST and ALT) elevation were grouped as those with acetaminophen exposure (AT) and those with elevated creatine phosphokinase (CPK) without evidence of acetaminophen exposure (RHB).Of the 345 patients included in the study, elevated AST/ALT levels were attributed to rhabdomyolysis in 168 patients and attributed to acetaminophen toxicity in 177 patients. The median AST: ALT values also differed between groups, with patients in the RHB group had higher median ratios (p 0.001). Using an AST: ALT value of 2.02 as a diagnostic cutoff produced a specificity of 0.52 (95% CI: 0.37, 0.64) and sensitivity of 0.84 (95% CI: 0.73, 0.94) for acetaminophen detection in the test dataset (N = 104).Elevated transaminases due to liver injury from acetaminophen ingestion had a different pattern than elevated transaminases due to rhabdomyolysis. Lower AST:ALT ratios were found in acetaminophen cases, however, the specificity using a ratio threshold of ≤1 would be 83%.

DNA methylation is an epigenetic mechanism controlling gene expression, and reduced methylation is associated with increased gene expression. We hypothesized that IL-17 cytokines are regulated by DNA methylation, are elevated in the circulation of preeclamptic women, and stimulate vascular neutrophil chemokine expression, which could account for vascular infiltration of neutrophils in preeclampsia. We found significantly reduced DNA methylation of IL17A, IL17E and IL17F genes in omental arteries of preeclamptic women, significantly reduced methylation of IL2, which regulates IL-17- producing T-lymphocytes, and significantly reduced methylation of genes encoding neutrophil chemokines and TNFα receptors related to lymphocyte function. Maternal plasma levels of IL-17A were significantly elevated in the second trimester of preeclamptic pregnancy as compared to normal pregnancy. To test if methylation regulates IL-17 cytokines, a lymphocyte cell line (Jurkat) was cultured with a hypomethylating agent. Hypomethylation increased expression of IL17E (aka IL25), IL17F and IL2. IL17A was not expressed by Jurkat cells. To test the potential role of IL-17 cytokines in vascular neutrophil infiltration associated with preeclampsia, human vascular smooth muscle cells were cultured with IL-17 cytokines. IL-17A, but not IL-17E or IL-17F, increased gene expression of neutrophil chemokines (IL-8, CXCL5 and CXCL6) that are increased in vascular smooth muscle of preeclamptic women. The monocyte chemokine, CCL-2, was not increased. TNFα also increased neutrophil chemokines. CONCLUSIONS: IL-17 cytokines are regulated by DNA methylation, IL-17A is elevated in preeclampsia and stimulates expression of neutrophil chemokines in vascular smooth muscle. SPECULATION: IL-17A could be responsible for vascular infiltration of neutrophils in preeclampsia.

Background A hallmark of Alzheimer’s disease is the accumulation of amyloid-β (Aβ) peptide. Donanemab, an antibody that targets a modified form of deposited Aβ, is being investigated for t...

Antibiotic resistance in bacterial pathogens is a growing problem for both human and veterinary medicine. Mobile genetic elements (MGEs) such as plasmids, transposons, and integrons enable the spread of antibiotic resistance genes (ARGs) among bacteria, and the overuse of antibiotics drives this process by providing the selection pressure for resistance genes to establish and persist in bacterial populations. Because bacteria, MGEs, and resistance genes can readily spread between different ecological compartments (e.g. soil, plants, animals, humans, wastewater), a “One Health” approach is needed to combat this problem. The equine hindgut is an understudied but potentially significant reservoir of ARGs and MGEs, since horses have close contact with humans, their manure is used in agriculture, they have a dense microbiome of both bacteria and fungi, and many antimicrobials used for equine treatment are also used in human medicine. Here, we collate information to date about resistance genes, plasmids, and class 1 integrons from equine-derived bacteria, we discuss why the equine hindgut deserves increased attention as a potential reservoir of ARGs, and we suggest ways to minimize the selection for ARGs in horses, in order to prevent their spread to the wider community.

Mitochondrial glycerol phosphate dehydrogenase (mGPD) is the rate-limiting enzyme of the glycerol phosphate redox shuttle. It was recently claimed that metformin, a first-line drug used for the treatment of type 2 diabetes, inhibits liver mGPD 30-50%, suppressing gluconeogenesis through a redox mechanism. Various factors cast doubt on this idea. Total-body knockout of mGPD in mice has adverse effects in several tissues where the mGPD level is high but has little or no effect in liver, where the mGPD level is the lowest of 10 tissues. Metformin has beneficial effects in humans in tissues with high levels of mGPD, such as pancreatic β-cells, where the mGPD level is much higher than that in liver. Insulin secretion in mGPD knockout mouse β-cells is normal because, like liver, β-cells possess the malate aspartate redox shuttle whose redox action is redundant to the glycerol phosphate shuttle. For these and other reasons, we used four different enzyme assays to reassess whether metformin inhibited mGPD. Metformin did not inhibit mGPD in homogenates or mitochondria from insulin cells or liver cells. If metformin actually inhibited mGPD, adverse effects in tissues where the level of mGPD is much higher than that in the liver could prevent the use of metformin as a diabetes medicine.

This work reports on the development of biomimetic polymeric cellulosomes—enzyme–polymer conjugates (EPCs) generated by covalent binding of cellulase enzymes onto a molecular brush polymer scaffold...

Familial hemophagocytic lymphohistiocytosis (HLH) is a life-threatening hyperinflammatory syndrome resulting from defective cytotoxicity. A previously healthy 3-month-old female presented with fever, irritability, abdominal distention, and tachypnea. She ultimately met all eight HLH-2004 diagnostic criteria, accompanied by elevated CXCL9. Initial empiric anti-inflammatory treatment included anakinra and IVIg, which stabilized ferritin and cytopenias. She had molecular and genetic confirmation of perforin deficiency and was started on dexamethasone and etoposide per HLH-94. She clinically improved, though CXCL9 and sIL-2Ra remained elevated. She was readmitted at week 8 for relapsed HLH without clear trigger and HLH-94 induction therapy was reinitiated. Her systemic HLH symptoms failed to respond and she soon developed symptomatic CNS HLH. She was incidentally found to have multifocal lung and kidney nodules, which were sterile and consisted largely of histiocytes and activated, oligoclonal CD8 T cells. The patient had a laboratory response to salvage therapy with alemtuzumab and emapalumab, but progressive neurologic decline led to withdrawal of care. This report highlights HLH foci manifest as pulmonary/renal nodules, demonstrates the utility of monitoring an array of HLH biomarkers, and suggests possible benefit of earlier salvage therapy.

Objectives To examine the correlation between spheno-occipital synchondrosis fusion stages and the hand-wrist skeletal maturity index. Materials and Methods Digital records of 164 individuals (77 males, 87 females) aged 10 to 18 years old were examined. Three-dimensional CBCT scans and hand-wrist two-dimensional radiographs were scored for the spheno-occipital synchondrosis fusion stages and hand-wrist skeletal maturity index, respectively. Statistical analyses were performed for associations using R software with a significance threshold of P< .01. Results A significant positive relationship was demonstrated between spheno-occipital synchondrosis fusion stages and hand-wrist skeletal maturity in both sexes. The Kendall's rank correlation τ between hand-wrist skeletal maturity index and spheno-occipital synchondrosis fusion percentage were high and positive in males and females (r = .74 and r = .71, respectively). Conclusions The significant, positive relationship between the hand-wrist skeletal maturity index and spheno-occipital synchondrosis fusion stages support the idea of using spheno-occipital synchondrosis fusion as a biological indicator for craniofacial and mandibular growth spurt prediction.

Objective To determine whether animals with Japanese macaque encephalomyelitis (JME), a spontaneous demyelinating disease similar to multiple sclerosis (MS), harbor myelin‐specific T cells in their central nervous system (CNS) and periphery. Methods Mononuclear cells (MNCs) from CNS lesions, cervical lymph nodes (LNs) and peripheral blood of Japanese macaques (JMs) with JME, and cervical LN and blood MNCs from healthy controls or animals with non‐JME conditions were analyzed for the presence of myelin‐specific T cells and changes in interleukin 17 (IL‐17) and interferon gamma (IFNγ) expression. Results Demyelinating JME lesions contained CD4+ T cells and CD8+ T cells specific to myelin oligodendrocyte glycoprotein (MOG), myelin basic protein (MBP), and/or proteolipid protein (PLP). CD8+ T‐cell responses were absent in JME peripheral blood, and in age‐ and sex‐matched controls. However, CD4+ Th1 and Th17 responses were detected in JME peripheral blood versus controls. Cervical LN MNCs from eight of nine JME animals had CD3+ T cells specific for MOG, MBP, and PLP that were not detected in controls. Mapping myelin epitopes revealed a heterogeneity in responses among JME animals. Comparison of myelin antigen sequences with those of JM rhadinovirus (JMRV), which is found in JME lesions, identified six viral open reading frames (ORFs) with similarities to myelin antigen sequences. Overlapping peptides to these JMRV ORFs did not induce IFNγ responses. Interpretations JME possesses an immune‐mediated component that involves both CD4+ and CD8+ T cells specific for myelin antigens. JME may shed new light on inflammatory demyelinating disease pathogenesis linked to gamma‐herpesvirus infection.

The University of Missouri (MU) has established a colony of dystrophin‐deficient dogs with a mixed breed background to mirror the variable pathologic effects of dystrophinopathies between persons of a given kindred to further the understanding of the genetic and molecular basis of the variable phenotype; thus to facilitate discovery of an effective therapeutic strategy. Herein we report the phenotype and genotype of a normal‐appearing 10‐month‐old colony female that died suddenly. At necropsy examination, there were reduced skeletal and laryngeal muscle volume and mild dilatation of the oesophagus. Microscopic findings consisted of extensive degeneration and regeneration of the axial skeletal, tongue, oesophageal, and laryngeal muscles that were characterized by considerable central nucleation, individual fibre mineralization and interstitial fibrosis. The myocardial findings were limited to infiltration of adipose cells in the interstitium. The female dog was a compound heterozygote with one X chromosome carrying a point mutation in intron 6 of the dystrophin gene and the other X chromosome carrying a repetitive element insertion in intron 13 of the dystrophin gene. Although the direct cause of death was uncertain, it might likely be due to sudden cardiac death as has been seen in Duchenne muscular dystrophy patients. This case demonstrated dystrophinopathy in female dogs that have no ameliorating normal X chromosome., A 10‐month‐old mixed breed female dog in a muscular dystrophy research colony at the University of Missouri died suddenly without premonitory symptoms. At necropsy examination, there were reduced skeletal and laryngeal muscle volume and mild dilatation of the esophagus. Microscopic findings consisted of extensive degeneration and regeneration of the axial skeletal, tongue, esophageal, and laryngeal muscles that were characterized by considerable central nucleation, individual fiber mineralization and interstitial fibrosis.

Despite optimal surgical resection, meningiomas may recur, with increasing grade and the degree of resection being predictive of risk. We hypothesize that an increasing Ki67 correlates with a higher risk of recurrence of resected WHO grade I meningiomas. The study population consisted of patients with resected WHO grade 1 meningiomas in locations outside of the base of skull. Digitally scanned slides stained for Ki67 were analyzed using automatic image analysis software in a standardized fashion. Recurrence was observed in 53 (17.7%) of cases with a median follow up time of 25.8 months. Ki67 ranged from 0 to 30%. Median Ki67 was 5.1% for patients with recurrence and 3.5% for patients without recurrence. In unadjusted analyses, high Ki-67 (≥ 5 vs. < 5) vs. ≥ 5) was associated with over a twofold increased risk of recurrence (13.1% vs. 27% respectively; HR 2.1731; 95% CI [1.2534, 3.764]; p = 0.006). After Adjusting for patient or tumor characteristics, elevated Ki-67 remained significantly correlated with recurrence. Grade 4 Simpson resection was noted in 71 (23.7%) of patients and it was associated with a significantly increased risk of recurrence (HR 2.56; 95% CI [1.41, 4.6364]; p = 0.002). WHO grade 1 meningiomas exhibit a significant rate of recurrence following resection. While Ki-67 is not part of the WHO grading criteria of meningiomas, a value greater than 5% is an independent predictor for increased risk of local recurrence following surgical resection.

Blood pressure is controlled by endocrine, autonomic, and behavioral responses that maintain blood volume and perfusion pressure at levels optimal for survival. Although it is clear that central angiotensin type 1a receptors (AT1aR; encoded by theAgtr1agene) influence these processes, the neuronal circuits mediating these effects are incompletely understood. The present studies characterize the structure and function of AT1aR neurons in the lamina terminalis (containing the median preoptic nucleus and organum vasculosum of the lamina terminalis), thereby evaluating their roles in blood pressure control. Using maleAgtr1a-Cre mice, neuroanatomical studies reveal that AT1aR neurons in the area are largely glutamatergic and send projections to the paraventricular nucleus of the hypothalamus (PVN) that appear to synapse onto vasopressin-synthesizing neurons. To evaluate the functionality of these lamina terminalis AT1aR neurons, we virally delivered light-sensitive opsins and then optogenetically excited or inhibited the neurons while evaluating cardiovascular parameters or fluid intake. Optogenetic excitation robustly elevated blood pressure, water intake, and sodium intake, while optogenetic inhibition produced the opposite effects. Intriguingly, optogenetic excitation of these AT1aR neurons of the lamina terminalis also resulted in Fos induction in vasopressin neurons within the PVN and supraoptic nucleus. Further, within the PVN, selective optogenetic stimulation of afferents that arise from these lamina terminalis AT1aR neurons induced glutamate release onto magnocellular neurons and was sufficient to increase blood pressure. These cardiovascular effects were attenuated by systemic pretreatment with a vasopressin-1a-receptor antagonist. Collectively, these data indicate that excitation of lamina terminalis AT1aR neurons induces neuroendocrine and behavioral responses that increase blood pressure.SIGNIFICANCE STATEMENTHypertension is a widespread health problem and risk factor for cardiovascular disease. Although treatments exist, a substantial percentage of patients suffer from “drug-resistant” hypertension, a condition associated with increased activation of brain angiotensin receptors, enhanced sympathetic nervous system activity, and elevated vasopressin levels. The present study highlights a role for angiotensin Type 1a receptor expressing neurons located within the lamina terminalis in regulating endocrine and behavioral responses that are involved in maintaining cardiovascular homeostasis. More specifically, data presented here reveal functional excitatory connections between angiotensin-sensitive neurons in the lamina terminals and vasopressin neurons in the paraventricular nucleus of the hypothalamus, and further indicate that activation of this circuit raises blood pressure. These neurons may be a promising target for antihypertensive therapeutics.

Quantifying predator–prey interactions and gaining insights into predator behavior are crucial for optimizing restoration strategies. However, such knowledge is often lacking for marine invertebrates. We examined potential impacts of predation by channeled Busycotypus canaliculatus and knobbed whelks Busycon carica on natural and planted populations of bay scallops in the Peconic Bays, New York, through laboratory and field investigations. In lab experiments, mean predation rates exhibited by small channeled whelks were low: 0.06 and 0.005 scallops d−1 for adult and juvenile scallops, respectively. Predation rates of small knobbed whelks on juvenile scallops were 22 × higher. Eighty-six percent (86%) of scallops consumed by channeled whelks had undamaged shells, while 73% eaten by knobbed whelks had notched ventral margins. In field plots where scallop densities were manipulated via removals/plantings, whelks consumed ~ 2% of ~ 19,100 planted juveniles, whereas crabs and presumably finfish consumed > 40% overall. Acoustic telemetry revealed that tagged channeled whelks moved shorter distances and spent more time in plots planted with scallops versus those without scallops. Whelks spent more time in low versus high-density plots, but consumed far more scallops in the latter. In trials without scallops, whelk movement rates were 5 × higher, presumably due to increased exploratory behavior. Overall, whelks were most active during crepuscular hours and during periods of increasing wind speeds. Our results, combined with population abundance data, suggest that whelks (especially B. carica) are drawn to planted bay scallop aggregations, but probably contribute to relatively low overall mortality in the context of restoration efforts.

Chemotherapy is used as a standard-of-care against cancers that display high levels of inherent genome instability. Chemotherapy induces DNA damage and intensifies pressure on the DNA repair pathways that can lead to deregulation. There is an urgent clinical need to be able to track the emergence of DNA repair driven chemotherapy resistance and tailor patient staging appropriately. There have been numerous studies into chemoresistance but to date no study has elucidated in detail the roles of the key DNA repair components in resistance associated with the frontline clinical combination of anthracyclines and taxanes together. In this study, we hypothesized that the emergence of chemotherapy resistance in triple negative breast cancer was driven by changes in functional signaling in the DNA repair pathways. We identified that consistent pressure on the non-homologous end joining pathway in the presence of genome instability causes failure of the key kinase DNA-PK, loss of p53 and compensation by p73. In-turn a switch to reliance on the homologous recombination pathway and RAD51 recombinase occurred to repair residual double strand DNA breaks. Further we demonstrate that RAD51 is an actionable target for resensitization to chemotherapy in resistant cells with a matched gene expression profile of resistance highlighted by homologous recombination in clinical samples.

Aquatic insects exhibit complex life cycles that include egg, larval, adult, and, in some instances, pupal stages. Disturbances at any of these life stages can affect overall population dyn...

Preclinical and clinical data suggest that acetyl-CoA carboxylase (ACC) inhibitors have the potential to rebalance disordered lipid metabolism, leading to improvements in nonalcoholic steatohepatitis (NASH). Consistent with these observations, first-in-human clinical trials with our ACC inhibitor PF-05175157 led to robust reduction of de novo lipogenesis (DNL), albeit with concomitant reductions in platelet count, which were attributed to the inhibition of fatty acid synthesis within bone marrow. Herein, we describe the design, synthesis, and evaluation of carboxylic acid-based ACC inhibitors with organic anion transporting polypeptide (OATP) substrate properties, which facilitated selective distribution of the compounds at the therapeutic site of action (liver) relative to the periphery. These efforts led to the discovery of clinical candidate PF-05221304 (12), which selectively inhibits liver DNL in animals, while demonstrating considerable safety margins against platelet reduction in a nonhuman primate model.

Abstract This study aims to compare the potential of Virginia mallow to other high yielding perennial grasses and hardwoods by characterising and comparing fast pyrolysis product yields. Feedstocks selected for this study include miscanthus (Miscanthus x giganteus), Virginia mallow (Sida hermaphrodita), willow short rotation coppice (SRC) (Salix viminalis) and oak (Quercus robur). The experimental work was split into two sections: analytical (Py–GC–MS) and laboratory-scale processing using a 300 g h−1 continuous bubbling fluidised bed reactor. Pyrolysis–gas chromatography–mass spectrometry (Py–GC–MS) has been used to quantify pyrolysis products from these feedstocks by simulating fast pyrolysis heating rates using a CDS 5200 pyrolyser closed coupled to a PerkinElmer Clarus 680 GC–MS. High bio-oil yields were achieved for Virginia mallow, willow SRC and oak (65.36, 62.55 and 66.43 wt% respectively), but miscanthus only produced a yield of 53.46 wt% due to increased feedstock ash content. The water content in the bio-oil is highest from miscanthus (17.64 wt%) and relatively low in the Virginia mallow and hardwoods willow SRC and oak (12.49, 13.88 and 14.53 wt%). Similar high yields of bio-oil and low yields of char and non-condensable gas compared to willow SRC make Virginia mallow an attractive feedstock for fast pyrolysis processing. Graphic Abstract

Understanding the dispersal ability of invasive insects provides useful insights for developing effective management strategies. Historically, methods for marking insects for dispersal studies have been expensive, time-consuming, and labor-intensive, especially for woodboring beetles. In addition, capturing or rearing insects requires human handling, which can alter behavior. Immunomarking is a well-established technique for studying the dispersal of insects, however, it has not been broadly applied to woodborers. This study evaluates the potential for using immunoproteins applied directly to woodborer-infested trees to mark emerging beetles. Specifically, in the first experiment I sprayed varying concentrations of ovalbumin (egg white) solution directly onto logs infested with emerald ash borer (EAB, Agrilus planipennis Fairmaire) (Buprestidae: Agrilini) and ELISA was used to detect the presence of protein on emerged beetles. To test the persistence of the mark, I applied varying concentrations of albumin to freeze-killed beetles, mounted them on pins, and placed them in an exposed location outdoors. Adult EAB self-marked as they emerged from protein-treated trees, with higher protein concentrations persisting for longer on the cuticle when exposed to sun and rain. This technique offers a convenient, inexpensive and durable means of marking woodborers and circumvents the need for human handling, allowing for more natural behavior and more realistic estimates of dispersal. Protein self-marking may find application in studies of woodborer dispersal within natural forest environments.

Detailed anatomical maps of individual organs and entire animals have served as invaluable entry points for ensuing dissection of their evolution, development, and function. The pharynx of the nematodeCaenorhabditis elegansis a simple neuromuscular organ with a self-contained, autonomously acting nervous system, composed of 20 neurons that fall into 14 anatomically distinct types. Using serial EM reconstruction, we re-evaluate here the connectome of the pharyngeal nervous system, providing a novel and more detailed view of its structure and predicted function. Contrasting the previous classification of pharyngeal neurons into distinct inter- and motorneuron classes, we provide evidence that most pharyngeal neurons are also likely sensory neurons and most, if not all, pharyngeal neurons also classify as motorneurons. Together with the extensive cross-connectivity among pharyngeal neurons, which is more widespread than previously realized, the sensory-motor characteristics of most neurons define a shallow network architecture of the pharyngeal connectome. Network analysis reveals that the patterns of neuronal connections are organized into putative computational modules that reflect the known functional domains of the pharynx. Compared to the somatic nervous system, pharyngeal neurons both physically associate with a larger fraction of their neighbors and create synapses with a greater proportion of their neighbors. We speculate that the overall architecture of the pharyngeal nervous system may be reminiscent of the architecture of ancestral, primitive nervous systems.

Epithelial plasticity, reversible modulation of a cell's epithelial and mesenchymal features, is associated with tumor metastasis and chemoresistance, leading causes of cancer mortality. Although different master transcription factors and epigenetic modifiers have been implicated in this process in various contexts, the extent to which a unifying, generalized mechanism of transcriptional regulation underlies epithelial plasticity remains largely unknown. Here, through targeted CRISPR/Cas9 screening, we discovered two histone-modifying enzymes involved in the writing and erasing of H3K36me2 that act reciprocally to regulate epithelial-to-mesenchymal identity, tumor differentiation, and metastasis. Using a lysine-to-methionine histone mutant to directly inhibit H3K36me2, we found that global modulation of the mark is a conserved mechanism underlying the mesenchymal state in various contexts. Mechanistically, regulation of H3K36me2 reprograms enhancers associated with master regulators of epithelial-to-mesenchymal state. Our results thus outline a unifying epigenome-scale mechanism by which a specific histone modification regulates cellular plasticity and metastasis in cancer. Significance: Although epithelial plasticity contributes to cancer metastasis and chemoresistance, no strategies exist for pharmacologically inhibiting the process. Here, we show that global regulation of a specific histone mark, H3K36me2, is a universal epigenome-wide mechanism that underlies epithelial-to-mesenchymal transition and mesenchymal-to-epithelial transition in carcinoma cells. These results offer a new strategy for targeting epithelial plasticity in cancer. This article is highlighted in the In This Issue feature, p. 747

Cancers develop as a result of driver mutations1,2 that lead to clonal outgrowth and the evolution of disease3,4. The discovery and functional characterization of individual driver mutations are central aims of cancer research, and have elucidated myriad phenotypes5 and therapeutic vulnerabilities6. However, the serial genetic evolution of mutant cancer genes7,8 and the allelic context in which they arise is poorly understood in both common and rare cancer genes and tumour types. Here we find that nearly one in four human tumours contains a composite mutation of a cancer-associated gene, defined as two or more nonsynonymous somatic mutations in the same gene and tumour. Composite mutations are enriched in specific genes, have an elevated rate of use of less-common hotspot mutations acquired in a chronology driven in part by oncogenic fitness, and arise in an allelic configuration that reflects context-specific selective pressures. cis-acting composite mutations are hypermorphic in some genes in which dosage effects predominate (such as TERT), whereas they lead to selection of function in other genes (such as TP53). Collectively, composite mutations are driver alterations that arise from context- and allele-specific selective pressures that are dependent in part on gene and mutation function, and which lead to complex-often neomorphic-functions of biological and therapeutic importance.

Bacteriophage T4 encodes orthologs of the proteins Rad50 (gp46) and Mre11 (gp47), which form a heterotetrameric complex (MR) that participates in the processing of DNA ends for recombination-dependent DNA repair. Crystal and high-resolution cryo-EM structures of Rad50 have revealed DNA binding sites near the dimer interface of Rad50 opposite of Mre11, and near the base of the coiled-coils that extend out from the globular head domain. An analysis of T4-Rad50 using sequenced-based algorithms to identify DNA binding residues predicts that a conserved region of positively charged residues near the C-terminus, distal to the observed binding sites, interacts with DNA. Mutant proteins were generated to test this prediction and their enzymatic and DNA binding activities were evaluated. Consistent with the predictions, the Rad50 C-terminal mutants had reduced affinity for DNA as measured by Rad50 equilibrium DNA binding assays and an increased Km-DNA as determined in MR complex nuclease assays. Moreover, the allosteric activation of ATP hydrolysis activity due to DNA binding was substantially reduced, suggesting that these residues may be involved in the communication between the DNA and ATP binding sites.

User-friendly computational tools for 16S ribosomal RNA (rRNA) sequencing analysis enable researchers who are not bioinformaticians to analyze and interpret sequencing data from microbial communities. These tools' easy-to-use interfaces belie the sophisticated and rapidly-evolving science of their underlying algorithms. When analyzing 16S data from a simple microbiome experiment, we found that superficially unimportant decisions about the bioinformatic pipeline led to results with radically different biological interpretations. We share these results as a cautionary tale whose moral is that, in 16S analysis, the devil is in the details. Wet bench researchers should therefore strongly consider partnering with bioinformaticians or computational biologists when analyzing 16S data.

Accumulating evidence in the literature suggests that oleic acid functions as a necromone across widely divergent insect taxa. The prevelance of this phenomenon has not been fully explored, and its application to pest management remains underdeveloped. Khapra beetle (KB), Trogoderma granarium, is a pest of stored grains, with larvae that can enter facultative diapause and remain cryptic in warehouses. Here, we examine how death affects oleic acid content of Trogoderma spp. cuticular extracts, and whether the compound causes a behavioral response. To assess the generalizability of patterns, many experiments were repeated with warehouse beetle (WB), Trogoderma variabile, and larger cabinet beetle (LCB), Trogoderma inclusum. Extracts of larvae that were first killed by being frozen had greater oleic acid content than those derived from live insects. Two-choice behavioral assays compared responses of solvent controls to these extracts, at both low (~ 2 µg) and high (68–131 µg) oleic acid content. The natural extracts also contained cuticular hydrocarbons and other unidentified chemicals. High oleic acid in the extracts repelled the larvae of all three species. Lower levels of oleic acid did not affect KB and LCB movement, but were attractive to WB. We also performed the assay using a large range of doses of oleic acid alone. At the lower doses, oleic acid had no effect on movement, but it became strongly repellant at higher doses, beginning at 100 µg. These results indicate that necromones may be an overlooked aspect of stored product insect biology, which if further researched could improve pest management.

Human immunodeficiency virus (HIV)-1 transactivator of transcription protein (Tat) is a viral protein that promotes transcription of the HIV genome and possesses cell-signaling properties. Long-term exposure of central nervous system (CNS) tissue to HIV-1 Tat is theorized to contribute to HIV-associated neurodegenerative disorder (HAND). In the current study, we sought to directly evaluate the effect of HIV-1 Tat expression on the intrinsic electrophysiological properties of pyramidal neurons located in layer 2/3 of the medial prefrontal cortex and in area CA1 of the hippocampus. Toward that end, we drove Tat expression with doxycycline (100 mg·kg−1·day−1 ip) in inducible Tat (iTat) transgenic mice for 7 days and then performed single-cell electrophysiological studies in acute tissue slices made through the prefrontal cortex and hippocampus. Control experiments were performed in doxycycline-treated G-tg mice, which retain the tetracycline-sensitive promoter but do not express Tat. Our results indicated that the predominant effects of HIV-1 Tat expression are excitatory in medial prefrontal cortical pyramidal neurons yet inhibitory in hippocampal pyramidal neurons. Notably, in these two populations, HIV-1 Tat expression produced differential effects on neuronal gain, membrane time constant, resting membrane potential, and rheobase. Similarly, we also observed distinct effects on action potential kinetics and afterhyperpolarization, as well as on the current-voltage relationship in subthreshold voltage ranges. Collectively, these data provide mechanistic evidence of complex and region-specific changes in neuronal physiology by which HIV-1 Tat protein may promote cognitive deficits associated with HAND. NEW & NOTEWORTHY We drove expression of human immunodeficiency virus (HIV)-1 transactivator of transcription protein (Tat) protein in inducible Tat (iTat) transgenic mice for 7 days and then examined the effects on the intrinsic electrophysiological properties of pyramidal neurons located in the medial prefrontal cortex (mPFC) and in the hippocampus. Our results reveal a variety of specific changes that promote increased intrinsic excitability of layer II/III mPFC pyramidal neurons and decreased intrinsic excitability of hippocampal CA1 pyramidal neurons, highlighting both cell type and region-specific effects.

Metabolic remodeling contributes to the development and progression of some cancers and exposes them to vulnerabilities, including specific nutrient dependencies that can be targeted therapeutically. Arginine is a semiessential amino acid, and several cancers are unable to endogenously synthesize sufficient levels of arginine for survival and proliferation, most commonly due to reduced expression of argininosuccinate synthase (ASS1). Such cancers are dependent on arginine and they can be targeted via enzyme-mediated depletion of systemic arginine. We report the preclinical safety, antitumor efficacy, and immune-potentiating effects of pegzilarginase, a highly potent human arginine-degrading enzyme. Toxicology studies showed that pegzilarginase-mediated arginine depletion is well tolerated at therapeutic levels that elicit an antitumor growth effect. To determine which tumor types are best suited for clinical development, we profiled clinical tumor samples for ASS1 expression, which correlated with pegzilarginase sensitivity in vivo in patient-derived xenograft (PDx) models. Among the histologies tested, malignant melanoma, small cell lung cancer and Merkel cell carcinoma had the highest prevalence of low ASS1 expression, the highest proportion of PDx models responding to pegzilarginase, and the strongest correlation between low or no ASS1 expression and sensitivity to pegzilarginase. In an immune-competent syngeneic mouse model, pegzilarginase slowed tumor growth and promoted the recruitment of CD8+ tumor infiltrating lymphocytes. This is consistent with the known autophagy-inducing effects of arginine depletion, and the link between autophagy and major histocompatibility complex antigen presentation to T cells. Our work supports the ongoing clinical investigations of pegzilarginase in solid tumors and clinical combination of pegzilarginase with immune checkpoint inhibitors.

The Anastrepha fraterculus (Wiedemann) complex is currently comprised of at least eight morphotypes, including several that are likely to be described as new species. It is critical to evaluate whether the morphotypes differ in tolerance to phytosanitary treatments. Temperatures from 0 to 3°C are used as a phytosanitary treatment for some commodities exported from the region and at risk of infestation by the A. fraterculus complex. Description of A. fraterculus morphotypes as new species could result in the annulation of phytosanitary treatment schedules for the new species. This study compared the relative cold tolerance of five populations from three morphotypes of the A. fraterculus complex: Andean, Peruvian, and Brazilian-1. Both a laboratory and wild strain of the Brazilian-1 morphotype were studied. Differences in mortality of third instars of the five A. fraterculus populations reared on nectarines were observed only with short treatment durations at temperatures ranging from 1.38 ± 0.04°C to 1.51 ± 0.08°C (mean ± SEM). Estimated times to achieve the LT99.99682 (probit 9) showed that Brazilian-1 wild, Brazilian-1 laboratory, and Cusco population were the most cold tolerant, followed by Andean and Peruvian, the least cold tolerant morphotype (i.e., Brazilian-1 wild = Brazilian-1 laboratory = Cusco population > Andean > Peruvian). These findings suggest that the current cold treatment schedules of 15 d at ≤ 1.11°C and 17 d at ≤ 1.67°C can be applied as cold treatments to any potential new species that may arise from the A. fraterculus complex.

Cancer genomics studies have identified thousands of putative cancer driver genes1. Development of high-throughput and accurate models to define the functions of these genes is a major challenge. Here we devised a scalable cancer-spheroid model and performed genome-wide CRISPR screens in 2D monolayers and 3D lung-cancer spheroids. CRISPR phenotypes in 3D more accurately recapitulated those of in vivo tumours, and genes with differential sensitivities between 2D and 3D conditions were highly enriched for genes that are mutated in lung cancers. These analyses also revealed drivers that are essential for cancer growth in 3D and in vivo, but not in 2D. Notably, we found that carboxypeptidase D is responsible for removal of a C-terminal RKRR motif2 from the α-chain of the insulin-like growth factor 1 receptor that is critical for receptor activity. Carboxypeptidase D expression correlates with patient outcomes in patients with lung cancer, and loss of carboxypeptidase D reduced tumour growth. Our results reveal key differences between 2D and 3D cancer models, and establish a generalizable strategy for performing CRISPR screens in spheroids to reveal cancer vulnerabilities. CRISPR screens in a 3D spheroid cancer model system more accurately recapitulate cancer phenotypes than existing 2D models and were used to identify carboxypeptidase D, acting via the IGF1R, as a 3D-specific driver of cancer growth.

Solid wood packaging material (WPM) is widely recognized as a high-risk pathway for transport and potential introduction of wood-boring insects, including longhorned beetles in the family Cerambycidae. These beetles also are occasionally imported in finished wood products, such as furniture and decorative items. A targeted effort to identify wood borers intercepted as larvae in WPM at U.S. ports between 2012 and 2018 revealed that one of the most frequently intercepted species was Trichoferus campestris (Faldermann), a cerambycid native to Asia. Trichoferus campestris is a pest of quarantine concern in the United States, Canada, and Europe. The establishment risk of this beetle in the United States is high because of its frequent introduction through multiple pathways and its potential to inhabit natural and urban forests as well as agricultural systems. In this study, we compiled port interception and detection data to examine risk based on historical introductions and pathways. We tested whether the intended destination of cargo intercepted with T. campestris-infested WPM can be used as a predictor of inland introductions, assuming that individuals of T. campestris are likely to be moved through established trade routes between export–import partners. We also developed maps to predict likely areas of introduction and establishment in the United States based on pathway analysis and climate suitability data. The maps will enable informed prioritization of resources in pest surveillance, and may serve as models for other wood borers identified in the WPM and wood products pathway.

Gene expression is extensively and dynamically modulated at the level of translation. How cancer cells prioritize the translation of certain mRNAs over others from a pool of competing mRNAs remains an open question. Here, we analyze translation in cell line models of breast cancer and normal mammary tissue by ribosome profiling. We identify key recurrent themes of oncogenic translation: higher ribosome occupancy, greater variance of translational efficiencies, and preferential translation of transcriptional regulators and signaling proteins in malignant cells as compared with their nonmalignant counterpart. We survey for candidate RNA interacting proteins that could associate with the 5′untranslated regions of the transcripts preferentially translated in breast tumour cells. We identify SRSF1, a prototypic splicing factor, to have a pervasive direct and indirect impact on translation. In a representative estrogen receptor–positive and estrogen receptor–negative cell line, we find that protein synthesis relies heavily on SRSF1. SRSF1 is predominantly intranuclear. Under certain conditions, SRSF1 translocates from the nucleus to the cytoplasm where it associates with MYC and CDK1 mRNAs and upregulates their internal ribosome entry site–mediated translation. Our results point to a synergy between splicing and translation and unveil how certain RNA-binding proteins modulate the translational landscape in breast cancer.

Metastasis-initiating cells with stem-like properties drive cancer lethality, yet their origins and relationship to primary-tumor-initiating stem cells are not known. We show that L1CAM+ cells in human colorectal cancer (CRC) have metastasis-initiating capacity, and we define their relationship to tissue regeneration. L1CAM is not expressed in the homeostatic intestinal epithelium, but is induced and required for epithelial regeneration following colitis and in CRC organoid growth. By using human tissues and mouse models, we show that L1CAM is dispensable for adenoma initiation but required for orthotopic carcinoma propagation, liver metastatic colonization and chemoresistance. L1CAMhigh cells partially overlap with LGR5high stem-like cells in human CRC organoids. Disruption of intercellular epithelial contacts causes E-cadherin–REST transcriptional derepression of L1CAM, switching chemoresistant CRC progenitors from an L1CAMlow to an L1CAMhigh state. Thus, L1CAM dependency emerges in regenerative intestinal cells when epithelial integrity is lost, a phenotype of wound healing deployed in metastasis-initiating cells. The authors identify a population of L1CAM-positive cells that, following loss of epithelial integrity, promote intestinal tissue regeneration and mediate metastasis initiation and chemoresistance in colorectal cancer.

Genetic variability of invasive pests informs us of fundamental aspects of the invasion, which help leverage limited resources for more efficient pest management. This is particularly important for managing species with invasive populations fueled by recurrent introductions. We present the first phylogeographic study of the velvet longhorned beetle (Trichoferus campestris [Faldermann]), a highly polyphagous wood-boring insect native to Asia. It attacks many ecologically and economically important tree species and has repeatedly invaded Europe and North America in recent decades. Despite the apparent threat posed by this pest, little work has been attempted to characterize genetic variability of T. campestris. One hurdle is the lack of genetic data beyond the “universal” COI barcode. Here we designed primer sequences to amplify five additional mitochondrial protein-coding genes for T. campestris (COII, COIII, Cytb, ND2, and ND4). We included specimens collected from domestic locations in the Unites States (U.S.) as well as samples intercepted in solid wood packaging material (WPM) at U.S. ports of entry. Phylogenetic analyses recovered three mitochondrial lineages, all including both domestic collections and port-interceptions. Our results demonstrated high levels of haplotype diversity among invasive populations of T. campestris. Specimens collected from the same geographical vicinity can be placed into different lineages. Species delimitation analyses further suggest presence of multiple molecular operational taxonomic units, which could complicate regulatory policies. We evaluated mechanisms that underlie the increasing number of T. campestris individuals and populations detected in the U.S. Results support the hypothesis of independent introductions through WPM, but are inconsistent with secondary invasions through the bridgehead effect.