COVID-19-Related Treatment Cancellations and Oncology Patients' Psychological Health in Nigeria.

Objective: To explore the association between COVID-19-related cancer treatment cancellations and the psychological health of cancer patients in Nigeria.Methods: We analyzed data collected from 15 outpatient cancer clinics, comprising 1,097 patients between April to July 2020. Study outcome was ten psychological impacts, including feeling down, stressed, and unable to access treatment due to COVID-19 (used as continuous and categorical variable (0-3,4-7,8+ events). The independent variable was treatment cancellations due to COVID-19 categorized as 0, 1, and 2+ cancellations. Confounders included religion, ethnicity, income, cancer diagnosis/type, and treatment received. Stata/SE.v.17 was used to perform all analyses. P values of ≤0.05 were deemed statistically significant.Results: Of the 1,097 cancer patients, 65.7% were female, with a mean age (SD) of 49.4 (13.8) years. Most patients (50.3%) reported four to seven psychological health events. Cancer patients who reported two/more treatment cancellations made up only 12.8% of the study sample but accounted for a greater proportion of psychological impacts (23.5%; P<0.001). In the adjusted model, cancer patients with one treatment cancellation (Coef: 0.195, 95%CI: 0.089-0.302) and those with two/more cancellations (Coef: 0.379, 95%CI: 0.255-0.504) had a significantly higher risk of psychological health impacts than those with no treatment cancellations.Conclusion: More than half of our sample of primarily adult female cancer patients reported major psychological health effects due to COVID-19. Cancer patients who experienced at least one treatment cancellation had a higher risk of psychological health consequences than those who did not. The implications of our findings and how to mitigate the impact of COVID-19 on oncology service disruptions are discussed.

Salicylic Acid Treatment Increases the Levels of Triterpene Glycosides in Black Cohosh (Actaea Racemosa) Rhizomes.

Black cohosh (Actaea racemosa) serves as the host plant for the Appalachian azure butterfly, Celastrina neglectamajor. Overharvesting of Black cohosh for the dietary supplement industry may result in its extirpation, and may also cause the elimination of the dependent butterfly. One way to increase or maintain the number of host plants in forested environments would be to reduce the number harvested, for example by increasing the levels of the desired metabolites in Black cohosh rhizomes. The secondary metabolites actein and deoxyactein are triterpene glycosides and are among the compounds associated with the putative activity of Black cohosh extracts. Acetein and deoxyacetein are used to standardize Black cohosh supplements. To gain an understanding of mechanisms that may control actein and deoxyactein accumulation, Black cohosh rhizomes were treated with exogenous salicylic acid, jasmonic acid, or ethylene, or were mechanically wounded. Salicylic acid treatment significantly increased the levels of actein and deoxyactein in the rhizome of Black cohosh, suggesting that the synthesis of triterpene glycosides is controlled in part by salicylic acid. Using salicylic acid or related chemicals to increase the levels of actein and deoxyactein in rhizomes may help supply the supplement industry and, simultaneously, help conserve Black cohosh and species dependent upon it.

The Potential of AI and ChatGPT in Improving Agricultural Injury and Illness Surveillance Programming and Dissemination.

Generative Artificial Intelligence (AI) provides unprecedented opportunities to improve injury surveillance systems in many ways, including the curation and publication of information related to agricultural injuries and illnesses. This editorial explores the feasibility and implication of ChatGPT integration in an international sentinel agricultural injury surveillance system, AgInjuryNews, highlighting that AI integration may enhance workflows by reducing human and financial resources and increasing outputs. In the coming years, text intensive natural language reports in AgInjuryNews and similar systems could be a rich source for data for ChatGPT or other more customized and fine-tuned LLMs. By harnessing the capabilities of AI and NLP, teams could potentially streamline the process of data analysis, report generation, and public dissemination, ultimately contributing to improved agricultural injury prevention efforts, well beyond any manually driven efforts.

The Landscape of Pediatric Radiation Oncology in Nigeria.

Radiation therapy (RT) is an essential part of the multidisciplinary treatment of pediatric cancer. Over the past five decades, significant advances have been made in the delivery of RT, with better dose delivery to disease targets while minimizing exposure to nearby organs at risk. These advances have led to improved treatment outcomes, increased survival, and reduced treatment-related toxicities. Advanced treatment techniques, however, require significant investment in infrastructural and personnel resources. This review documents what is currently available regarding expertise and infrastructure for pediatric radiation oncology practice in Nigeria. It was performed to serve as a foundation for the creation and design of tailored solutions (initiatives and policies) to increase pediatric radiation availability, accessibility, and equity in Nigeria and ultimately improve pediatric cancer treatment outcomes in the region.

Feasibility and Safety of Implementing Volumetric Arc Therapy (VMAT) for Pediatric Craniospinal Irradiation in a Low-Middle-Income Region: The Nigerian Experience.

Purpose: Volumetric modulated arc therapy (VMAT) is a relatively new treatment technique in sub-Saharan Africa. Although craniospinal irradiation (CSI) in the pediatric population has been practiced in Nigeria for many years, the use of VMAT to deliver this treatment is previously undocumented. We reviewed the first set of patients to undergo CSI at a cancer center in Nigeria, detailing the treatment technique, the progress experienced, dose statistics achieved, treatment toxicities, and cancer outcomes to date. Methods and Materials: This was a prospective case series of 5 children with histologically diagnosed cancers requiring CSI whose parents consented to the study. They were recruited at evaluation and followed through the process of their therapy. Toxicity was monitored at weekly review appointments using the Common Terminology Criteria for Adverse Events version 5.0. Follow-up of the children will continue in the long-term effects clinic. Results: Five patients with a median age of 6 were recruited. Diagnoses were intracranial germ cell tumor (n = 2), medulloblastoma (n = 1), pineoblastoma (n = 1), and ependymoma (n = 1). For all patients, a dose of 36.0 Gy in 1.8 Gy daily fractions was prescribed to the entire neuraxis. A subsequent boost of 18 Gy (n = 4) to 19.8 Gy (n = 1) in 10 daily fractions to the primary tumor bed (n = 2) and posterior fossa (n = 2) was delivered. Four patients had chemotherapy before, during, or after radiation therapy. No patient experienced grade 3 or greater toxicity. Conclusions: Our results indicate great progress has been made in the delivery of CSI in Nigeria, demonstrating tolerable acute side effects using VMAT. This series suggests the feasibility of implementing VMAT technology in low- or middle-income countries. Additional follow-up will be needed to determine whether survival rates and chronic toxicity rates are similar to those reported in the literature.

Evaluating the adoption of voice recognition technology for real-time dictation in a rural healthcare system: A retrospective analysis of dragon medical one.

BACKGROUND: In 2013, Marshfield Clinic Health System (MCHS) implemented the Dragon Medical One (DMO) system provided by Nuance Management Center (NMC) for Real-Time Dictation (RTD), embracing the idea of streamlined clinic workflow, reduced dictation hours, and improved documentation legibility. Since then, MCHS has observed a trend of reduced time in documentation, however, the target goal of 100% adoption of voice recognition (VR)-based RTD has not been met. OBJECTIVE: To evaluate the uptake/adoption of VR technology for RTD in MCHS, between 2018-2020. METHODS: DMO data for 1,373 MCHS providers from 2018-2020 were analyzed. The study outcome was VR uptake, defined as the median number of hours each provider used VR technology to dictate patient information, and classified as no/yes. Covariates included sex, age, US-trained/international medical graduates, trend, specialty, and facility. Descriptive statistics and unadjusted and adjusted logistic regression analyses were performed. Stata/SE.version.17 was used for analyses. P-values less than/equal to 0.05 were considered statistically significant. RESULTS: Of the 1,373 MCHS providers, the mean (SD) age was 48.3 (12.4) years. VR uptake was higher than no uptake (72.0% vs. 28.0%). In both unadjusted and adjusted analyses, VR uptake was 4.3 times and 7.7 times higher in 2019-2020 compared to 2018, respectively (OR:4.30,95%CI:2.44-7.46 and AOR:7.74,95%CI:2.51-23.86). VR uptake was 0.5 and 0.6 times lower among US-trained physicians compared to internationally-trained physicians (OR:0.53,95%CI:0.37-0.76 and AOR:0.58,95%CI:0.35-0.97). Uptake was 0.2 times lower among physicians aged 60/above than physicians aged 29/less (OR:0.20,95%CI:0.10-0.59, and AOR:0.17,95%CI:0.27-1.06). CONCLUSION: Since 2018, VR adoption has increased significantly across MCHS. However, it was lower among US-trained physicians than among internationally-trained physicians (although internationally physicians were in minority) and lower among more senior physicians than among younger physicians. These findings provide critical information about VR trends, physician factors, and which providers could benefit from additional training to increase VR adoption in healthcare systems.

Your neighborhood matters: an ecological social determinant study of the relationship between residential racial segregation and the risk of firearm fatalities.

BACKGROUND: Firearm fatalities are a major public health concern, claiming the lives of 40,000 Americans each year. While firearm fatalities have pervasive effects, it is unclear how social determinants of health (SDOH) such as residential racial segregation, income inequality, and community resilience impact firearm fatalities. This study investigates the relationships between these SDOH and the likelihood of firearm fatalities. METHODS: County-level SDOH data from the Agency for Health Care Research and Quality for 2019 were analyzed, covering 72 Wisconsin counties. The dependent variable was the number of firearm fatalities in each county, used as a continuous variable. The independent variable was residential racial segregation (Dissimilarity Index), defined as the degree to which non-White and White residents were distributed across counties, ranging from 0 (complete integration) to 100 (complete segregation), and higher values indicate greater residential segregation (categorized as low, moderate, and high). Covariates were income inequality ranging from zero (perfect equality) to one (perfect inequality) categorized as low, moderate, and high, community resilience risk factors (low, moderate, and high risks), and rural-urban classifications. Descriptive/summary statistics, unadjusted and adjusted negative binomial regression adjusting for population weight, were performed using STATA/MPv.17.0; P-values ≤ 0.05 were considered statistically significant. ArcMap was used for Geographic Information System analysis. RESULTS: In 2019, there were 802 firearm fatalities. The adjusted model demonstrates that the risk of firearm fatalities was higher in areas with high residential racial segregation compared to low-segregated areas (IRR.:1.26, 95% CI:1.04-1.52) and higher in areas with high-income inequality compared to areas with low-income inequality (IRR.:1.18, 95% CI:1.00-1.40). Compared to areas with low-risk community resilience, the risk of firearm fatalities was higher in areas with moderate (IRR.:0.61, 95% CI:0.48-0.78), and in areas with high risk (IRR.:0.53, 95% CI:0.41-0.68). GIS analysis demonstrated that areas with high racial segregation also have high rates of firearm fatalities. CONCLUSION: Areas with high residential racial segregation have a high rate of firearm fatalities. With high income inequality and low community resilience, the likelihood of firearm fatalities increases.

Examining Cancer Patients' Perceptions of the Impact of COVID-19 on Teleoncology: Findings From 15 Nigerian Outpatient Cancer Clinics.

PURPOSE: To examine cancer patients' perspectives on the impact of COVID-19 on teleoncology in Nigeria. METHODS: Data from a multicenter survey conducted at 15 outpatient clinics to 1,097 patients with cancer from April and July 2020 were analyzed. The study outcome was telemedicine, defined as patients who reported their routine follow-up visits were converted to virtual visits because of COVID-19 (coded yes/no). Covariates included patient age, ethnicity, marital status, income, cancer treatment, service disruption, and cancer diagnosis/type. Stata/SE.v.17 (StataCorp, College Station, TX) was used to perform chi-square and logistic regression analyses. P values ≤ .05 were considered statistically significant. RESULTS: The majority of the 1,097 patients with cancer were female (65.7%) and age 55 years and older (35.0%). Because of COVID-19, 12.6% of patients' routine follow-ups were converted to virtual visits. More patients who canceled/postponed surgery (17.7% v 7.5%; P ≤ .001), radiotherapy (16.9% v 5.3%; P ≤ .001), and chemotherapy (22.8% v 8.5%; P ≤ .001), injection chemotherapy (20.6% v 8.7%; P ≤ .001) and those who reported being seen less by their doctor/nurse (60.3% v 11.4%; P ≤ .001) reported more follow-up conversions to virtual visits. In multivariate analyses, patients seen less by their doctors/nurses were 14.3 times more likely to have their routine follow-ups converted to virtual visits than those who did not (odds ratio, 14.33; 95% CI, 8.36 to 24.58). CONCLUSION: COVID-19 caused many patients with cancer in Nigeria to convert visits to a virtual format. These conversions were more common in patients whose surgery, radiotherapy, chemotherapy, and injection chemotherapy treatments were canceled or postponed. Our findings suggest how COVID-19 affects cancer treatment services and the importance of collecting teleoncological care data in Nigeria.

Fly-CURE, a Multi-institutional CURE using Drosophila, Increases Students' Confidence, Sense of Belonging, and Persistence in Research.

The Fly-CURE is a genetics-focused multi-institutional Course-Based Undergraduate Research Experience (CURE) that provides undergraduate students with hands-on research experiences within a course. Through the Fly-CURE, undergraduate students at diverse types of higher education institutions across the United States map and characterize novel mutants isolated from a genetic screen in Drosophila melanogaster. To evaluate the impact of the Fly-CURE experience on students, we developed and validated assessment tools to identify students' perceived research self-efficacy, sense of belonging in science, and intent to pursue additional research opportunities. Our data show gains in these metrics after completion of the Fly-CURE across all student subgroups analyzed, including comparisons of gender, academic status, racial and ethnic groups, and parents' educational background. Importantly, our data also show differential gains in the areas of self-efficacy and interest in seeking additional research opportunities between Fly-CURE students with and without prior research experience, illustrating the positive impact of research exposure (dosage) on student outcomes. Altogether, our data indicate that the Fly-CURE experience has a significant impact on students' efficacy with research methods, sense of belonging to the scientific community, and interest in pursuing additional research experiences.

Fly-CURE, a multi-institutional CURE using Drosophila, increases students' confidence, sense of belonging, and persistence in research.

The Fly-CURE is a genetics-focused multi-institutional Course-Based Undergraduate Research Experience (CURE) that provides undergraduate students with hands-on research experiences within a course. Through the Fly-CURE, undergraduate students at diverse types of higher education institutions across the United States map and characterize novel mutants isolated from a genetic screen in Drosophila melanogaster. To date, more than 20 mutants have been studied across 20 institutions, and our scientific data have led to eleven publications with more than 500 students as authors. To evaluate the impact of the Fly-CURE experience on students, we developed and validated assessment tools to identify students' perceived research self-efficacy, sense of belonging in science, and intent to pursue additional research opportunities. Our data, collected over three academic years and involving 14 institutions and 480 students, show gains in these metrics after completion of the Fly-CURE across all student subgroups analyzed, including comparisons of gender, academic status, racial and ethnic groups, and parents' educational background. Importantly, our data also show differential gains in the areas of self-efficacy and interest in seeking additional research opportunities between Fly-CURE students with and without prior research experience, illustrating the positive impact of research exposure (dosage) on student outcomes. Altogether, our data indicate that the Fly-CURE experience has a significant impact on students' efficacy with research methods, sense of belonging to the scientific research community, and interest in pursuing additional research experiences.

Contents of Cannabinoids in Hemp Varieties Grown in Maryland.

Coincident with the cannabis legalization and the increased interest in the medicinal use of the plant, the cannabis marketplace and farming have seen tremendous growth. It is reported that there are more than 2000 cannabis varieties available to customers. However, the data that is available to the growers and breeders regarding the cannabinoid contents of various varieties remains low. Here, a high-performance liquid chromatography (HPLC) method was developed and validated for the simultaneous separation and determination of 11 cannabinoids. A total of 104 hemp bud materials belonging to 20 varieties were collected from farms in the state of Maryland and analyzed with the HPLC method. The contents of the 11 cannabinoids in various varieties were compared and discussed, highlighting the varieties that showed a high yield of cannabinoids and good consistency that are more appropriate for cannabinoid production.

Tomato pathogenesis-related protein genes are expressed in response to Trialeurodes vaporariorum and Bemisia tabaci biotype B feeding.

The temporal and spatial expression of tomato wound- and defense-response genes to Bemisia tabaci biotype B (the silverleaf whitefly) and Trialeurodes vaporariorum (the greenhouse whitefly) feeding were characterized. Both species of whiteflies evoked similar changes in tomato gene expression. The levels of RNAs for the methyl jasmonic acid (MeJA)- or ethylene-regulated genes that encode the basic ß-1,3-glucanase (GluB), basic chitinase (Chi9), and Pathogenesis-related protein-1 (PR-1) were monitored. GluB and Chi9 RNAs were abundant in infested leaves from the time nymphs initiated feeding (day 5). In addition, GluB RNAs accumulated in apical non-infested leaves. PR-1 RNAs also accumulated after whitefly feeding. In contrast, the ethylene- and salicylic acid (SA)-regulated Chi3 and PR-4 genes had RNAs that accumulated at low levels and GluAC RNAs that were undetectable in whitefly-infested tomato leaves. The changes in Phenylalanine ammonia lyase5 (PAL5) were variable; in some, but not all infestations, PAL5 RNAs increased in response to whitefly feeding. PAL5 RNA levels increased in response to MeJA, ethylene, and abscisic acid, and declined in response to SA. Transcripts from the wound-response genes, leucine aminopeptidase (LapA1) and proteinase inhibitor 2 (pin2), were not detected following whitefly feeding. Furthermore, whitefly infestation of transgenic LapA1:GUS tomato plants showed that whitefly feeding did not activate the LapA1 promoter, although crushing of the leaf lamina increased GUS activity up to 40 fold. These studies indicate that tomato plants perceive B. tabaci and T. vaporariorum in a manner similar to baterical pathogens and distinct from tissue-damaging insects.

GmEREBP1 is a transcription factor activating defense genes in soybean and Arabidopsis.

Ethylene-responsive element-binding proteins (EREBPs) are plant-specific transcription factors, many of which have been linked to plant defense responses. Conserved EREBP domains bind to the GCC box, a promoter element found in pathogenesis-related (PR) genes. We previously identified an EREBP gene from soybean (GmEREBP1) whose transcript abundance decreased in soybean cyst-nematode-infected roots of a susceptible cultivar, whereas it increased in abundance in infected roots of a resistant cultivar. Here, we report further characterization of this gene. Transient expression analyses showed that GmEREBP1 is localized to the plant nucleus and functions as a transcriptional activator in soybean leaves. Transgenic soybean plants expressing GmEREBP1 activated the expression of the ethylene (ET)-responsive gene PR2 and the ET- and jasmonic acid (JA)-responsive gene PR3, and the salicylic acid (SA)-responsive gene PR1 but not the SA-responsive PR5. Similarly, transgenic Arabidopsis plants expressing GmEREBP1 showed elevated mRNA abundance of the ET-regulated gene PR3 and the ET- and JA-regulated defense-related gene PDF1.2 but not the ET-regulated GST2, and the SA-regulated gene PR1 but not the SA-regulated PR2 and PR5. Transgenic soybean and Arabidopsis plants inoculated with cyst nematodes did not display a significantly altered susceptibility to nematode infection. These results collectively show that GmEREBP1 functions as a transacting inducer of defense gene expression in both soybean and Arabidopsis and mediates the expression of both ET- and JA- and SA-regulated defense-related genes in these plant species.

Gene-for-gene defense of wheat against the Hessian fly lacks a classical oxidative burst.

Genetic similarities between plant interactions with microbial pathogens and wheat interactions with Hessian fly larvae prompted us to investigate defense and counterdefense mechanisms. Plant oxidative burst, a rapid increase in the levels of active oxygen species (AOS) within the initial 24 h of an interaction with pathogens, commonly is associated with defenses that are triggered by gene-for-gene recognition events similar to those involving wheat and Hessian fly larvae. RNAs encoded by Hessian fly superoxide dismutase (SOD) and catalase (CAT) genes, involved in detoxification of AOS, increased in first-instar larvae during both compatible and incompatible interactions. However, mRNA levels of a wheat NADPH oxidase (NOX) gene that generates superoxide (O2-) did not increase. In addition, inhibiting wheat NOX enzyme with diphenyleneiodonium did not result in increased survival of avirulent larvae. However, nitro blue tetrazolium staining indicated that basal levels of O2- are present in both uninfested and infested wheat tissue. mRNA encoded by wheat genes involved in detoxification of the cellular environment, SOD, CAT, and glutathione-S-transferase did not increase in abundance. Histochemical staining with 3,3-diaminobenzidine revealed no increases in wheat hydrogen peroxide (H2O2) during infestation that were correlated with the changes in larval SOD and CAT mRNA. However, treatment with 2',7'-dichlorofluorescin demonstrated the presence of basal levels of H2O2 in the elongation zone of both infested and uninfested plants. The accumulation of a wheat flavanone 3-hydroxylase mRNA did show some parallels with larval gene mRNA profiles. These results suggested that larvae encounter stresses imposed by mechanisms other than an oxidative burst in wheat seedlings.

Identification and characterization of a soybean ethylene-responsive element-binding protein gene whose mRNA expression changes during soybean cyst nematode infection.

Ethylene-responsive element-binding proteins (EREBPs) are members of a family of plant transcription factors. Conserved EREBP domains of these proteins bind to the GCC box, an ethylene-responsive promoter element found in many pathogenesis-related (PR) genes. Using degenerate primers to the EREBP domain from diverse plant species, an EREBP homolog was isolated from a soybean cDNA library. Gel mobility-shift assays revealed that the translation product of this cDNA bound specifically to GCC box sequences. We, therefore, named this gene Glycine max ethylene-responsive element-binding protein 1 (GmEREBP1), i.e., a gene coding for the first confirmed GCC box-binding protein of soybean. GmEREBP1 mRNA abundance was analyzed by RNA blot hybridizations in soybean roots and shoots of cultivars Corsoy 79 and Hartwig, which are susceptible and resistant, respectively, to the soybean cyst nematode (Heterodera glycines). These analyses revealed that GmEREBP1 is expressed in a root-preferential manner and that GmEREBP1 mRNA abundance is changed after H. glycines infection. GmEREBP1 mRNA abundance decreased in infected (susceptible) 'Corsoy 79' roots, whereas it increased in abundance in infected (resistant) 'Hartwig' roots. Furthermore, ethephon treatment repressed GmEREBP1 mRNA accumulation in both cultivars, whereas wounding increased expression in both cultivars. These changes in mRNA steady-state levels suggest that GmEREBP1 plays a role in soybean-H. glycines interactions.

Insect feeding-induced differential expression of Beta vulgaris root genes and their regulation by defense-associated signals.

Root responses to insect pests are an area of plant defense research that lacks much information. We have identified more than 150 sugar beet root ESTs enriched for genes responding to sugar beet root maggot feeding from both moderately resistant, F1016, and susceptible, F1010, genotypes using suppressive subtractive hybridization. The largest number of identified F1016 genes grouped into the defense/stress response (28%) and secondary metabolism (10%) categories with a polyphenol oxidase gene, from F1016, identified most often from the subtractive libraries. The differential expression of the root ESTs was confirmed with RT-PCR. The ESTs were further characterized using macroarray-generated expression profiles from F1016 sugar beet roots following mechanical wounding and treatment of roots with the signaling molecules methyl jasmonate, salicylic acid and ethylene. Of the examined root ESTs, 20, 17 and 11% were regulated by methyl jasmonate, salicylic acid and ethylene, respectively, suggesting these signaling pathways are involved in sugar beet root defense responses to insects. Identification of these sugar beet root ESTs provides knowledge in the field of plant root defense and will lead to the development of novel control strategies for control of the sugar beet root maggot.

GeneChip profiling of transcriptional responses to soybean cyst nematode, Heterodera glycines, colonization of soybean roots.

Soybean cyst nematode (SCN) is currently the most devastating pathogen of soybean. SCN penetrates the root and migrates toward the central vascular bundle where it establishes a complex multinucleated feeding structure that provides plant-derived nutrients to support the development and growth of the nematode. To identify host genes that play significant roles in SCN development in susceptible roots, RNA from SCN-inoculated and non-inoculated root pieces were hybridized to the Affymetrix soybean genome GeneChips. RNA was collected at 8, 12, and 16 d post-inoculation from root pieces that displayed multiple swollen female SCN and similar root pieces from non-inoculated roots. Branch roots and root tips were trimmed from the root pieces to minimize the amount of RNA contributed by these organs. Of the 35 593 transcripts represented on the GeneChip, approximately 26,500 were expressed in the SCN-colonized root pieces. ANOVA followed by False Discovery Rate analysis indicated that the expression levels of 4616 transcripts changed significantly (Q-value < or =0.05) in response to SCN. In this set of 4616 transcripts, 1404 transcripts increased >2-fold and 739 decreased >2-fold. Of the transcripts to which a function could be assigned, a large proportion was associated with cell wall structure. Other functional categories that included a large number of up-regulated transcripts were defence, metabolism, and histones, and a smaller group of transcripts associated with signal transduction and transcription.

A novel wheat gene encoding a putative chitin-binding lectin is associated with resistance against Hessian fly.

SUMMARY The gene-for-gene interaction triggering resistance of wheat against first-instar Hessian fly larvae utilizes specialized defence response genes not previously identified in other interactions with pests or pathogens. We characterized the expression of Hfr-3, a novel gene encoding a lectin-like protein with 68-70% identity to the wheat germ agglutinins. Within each of the four predicted chitin-binding hevein domains, the HFR-3 translated protein sequence contained five conserved saccharide-binding amino acids. Quantification of Hfr-3 mRNA levels confirmed a rapid response and gradual increase, up to 3000-fold above the uninfested control in the incompatible interaction 3 days after egg hatch. Hfr-3 mRNA abundance was influenced by the number of larvae per plant, suggesting that resistance is localized rather than systemic. In addition, Hfr-3 was responsive to another sucking insect, the bird cherry-oat aphid, but not to fall armyworm attack, wounding or exogenous application of methyl jasmonate, salicylic acid or abscisic acid. Western blot analysis demonstrated that HFR-3 protein increased in parallel to mRNA levels in crown tissues during incompatible interactions. HFR-3 protein was detected in both virulent and avirulent larvae, indicating ingestion. Anti-nutritional proteins, such as lectins, may be responsible for the apparent starvation of avirulent first-instar Hessian fly larvae during the initial few days of incompatible interactions with resistant wheat plants.

Hfr-2, a wheat cytolytic toxin-like gene, is up-regulated by virulent Hessian fly larval feedingdouble dagger.

SUMMARY Both yield and grain-quality are dramatically decreased when susceptible wheat (Triticum aestivum) plants are infested by Hessian fly (Mayetiola destructor) larvae. Examination of the changes in wheat gene expression during infestation by virulent Hessian fly larvae has identified the up-regulation of a gene, Hessian fly responsive-2 (Hfr-2), which contains regions similar to genes encoding seed-specific agglutinin proteins from Amaranthus. Hfr-2, however, did not accumulate in developing seeds, as do other wheat seed storage proteins. Additionally, a separate region of the HFR-2 predicted amino acid sequence is similar to haemolytic proteins, from both mushroom and bacteria, that are able to form pores in cell membranes of mammalian red blood cells. The involvement of Hfr-2 in interactions with insects was supported by experiments demonstrating its up-regulation by both fall armyworm (Spodoptera frugiperda) and bird cherry-oat aphid (Rhopalosiphum padi) infestations but not by virus infection. Examination of wheat defence response pathways showed Hfr-2 up-regulation following methyl jasmonate treatment and only slight up-regulation in response to salicylic acid, abscisic acid and wounding treatments. Like related proteins, HFR-2 may normally function in defence against certain insects or pathogens. However, we propose that as virulent Hessian fly larvae manipulate the physiology of the susceptible host, the HFR-2 protein inserts in plant cell membranes at the feeding sites and by forming pores provides water, ions and other small nutritive molecules to the developing larvae.

ATP-dependent hexameric assembly of the heat shock protein Hsp101 involves multiple interaction domains and a functional C-proximal nucleotide-binding domain.

Members of the Hsp100 family of heat stress proteins are present in species throughout the bacterial, plant, and fungal kingdoms. Most Hsp100 proteins are composed of five domains that include two nucleotide-binding domains required for their ATP-dependent oligomerization. Mutations within the first but not the second nucleotide-binding site disrupt self-assembly of bacterial Hsp100, whereas the reverse is true for yeast Hsp104. We have examined the functional requirements for oligomerization of plant Hsp101 and have found that Hsp101 resembles Hsp104 in that it assembles into a hexameric complex in an ATP-dependent manner. Self-assembly of Hsp101 involves at least three distinct interaction domains located in the N-proximal domain and in the first and second nucleotide-binding domains. The interaction domain in the second nucleotide-binding domain included the Walker A motif, and mutations within this element disrupted self-assembly of Hsp101. In contrast, mutations affecting conserved residues of the Walker A motif within the first nucleotide-binding site did not affect self-assembly. No interaction between Hsp101 and Hsp104 was observed. These results suggest that plant Hsp101 self-assembly involves multiple evolutionarily diverged interaction domains as well as an evolutionarily conserved requirement for a functional C-proximal nucleotide-binding site.