Bulk segregant mapping and transcriptome analyses reveal the molecular mechanisms of spinetoram resistance in Spodoptera frugiperda.

The evolution of resistance to insecticides poses a significant threat to pest management programs. Understanding the molecular mechanisms underlying insecticide resistance is essential to design sustainable pest control and resistance management programs. The fall armyworm, Spodoptera frugiperda, is an important insect pest of many crops and has a remarkable ability to evolve resistance to insecticides. In this study, we employed bulk segregant analysis (BSA) combined with DNA and RNA sequencing to characterize the molecular basis of spinetoram resistance in S. frugiperda. Analysis of genomic data derived from spinetoram selected and unselected bulks and the spinetoram-resistant and susceptible parental strains led to the identification of a three-nucleotide deletion in the gene encoding the nicotinic acetylcholine receptor α6 subunit (nAChR α6). Transcriptome profiling identified the upregulation of few genes encoding detoxification enzymes associated with spinetoram resistance. Thus, spinetoram resistance in S. frugiperda appears to be mediated mainly by target site insensitivity with a minor role of detoxification enzymes. Our findings provide insight into the mechanisms underpinning resistance to spinetoram in S. frugiperda and will inform the development of strategies to control this highly damaging, globally distributed crop pest.

Responses of two Anastrepha species' immature stages infesting preferential hosts to different temperature exposures.

Anastrepha fraterculus (Wiedemann) and A. obliqua (Macquart) are important pests of fruit crops. In Brazil, these species cause damage to fruit growing in the South (annual average temperature of 20.9 °C) and Northeast (average yearly temperature of 24 °C). We evaluated the effect of temperature on the viability and development time of A. fraterculus and A. obliqua immature stages in their respective preferred hosts, guava (Psidium guajava L., Myrtaceae) and mango (Mangifera indica L., Anacardiaceae). The duration of egg and pupal stages, egg to pre-pupa, and viability of egg and pupal stages under different temperatures (15, 20, 25, 30, and 35 °C) were assessed. For both species, development time decreased with increasing temperature. Viability in the evaluated stages was only observed between 15 and 30 °C. However, the species responded differently to the exposure temperatures (15 and 30 °C), especially in the pupal stage and from egg to pre-pupa. Anastrepha fraterculus showed a lower tolerance to high temperatures, especially in the pupal stage and from egg to pre-pupa, which may explain its lower importance and economic impact in warmer Brazilian regions. Anastrepha obliqua had a lower tolerance at 15 °C, indicating greater adequacy for temperatures above 20 °C, characteristic of Northeast Brazil, suggesting the capacity to spread to cooler areas with rising temperatures.

Gingival metastasis of angiosarcoma of the breast as a first manifestation of spreading disease: Case report and review of the literature.

BACKGROUND: Angiosarcoma is an aggressive malignant neoplasm of vascular origin. Oral metastases of angiosarcoma are rare and have a non-specific clinical presentation, thus the diagnosis may be challenging. CASE REPORT: Herein we report a case of a 34-year-old female patient after treatment of a high-grade angiosarcoma of the breast, who presented an asymptomatic bleeding purplish nodule in the maxillary interdental papilla between the first and second premolar. A biopsy was performed, and the histological examination revealed infiltration by malignant neoplasm of epithelioid and fusocellular pattern. Immunohistochemical analysis demonstrated that neoplastic cells were positive for ERG and CD31, and negative for cytokeratins AE1/AE3, confirming the diagnosis of metastatic angiosarcoma. After investigation, multiple metastases were discovered. The patient is under management with chemotherapy and palliative radiotherapy for the bone lesions. CONCLUSION: Metastases should be considered in the differential diagnosis of oral lesions in patients with a previous history of cancer. Due to the morphology of angiosarcomas, the metastatic lesions may resemble benign vascular lesions, therefore, biopsy is mandatory to exclude malignancy.

Uterine Sarcoma With EML4::NTRK3 Fusion: A Spectrum of Mesenchymal Neoplasms Harboring Actionable Gene Fusions.

NTRK gene fusions are part of a paradigm shift in oncology, arising as one of the main genomic alterations with actionability in the so-called "agnostic setting." In gynecologic pathology, the recent description of uterine sarcoma resembling fibrosarcoma and with NTRK rearrangements ( NTRK -rearranged uterine sarcoma) highlights the importance of recognizing clinicopathological cues that can lead to genomic profiling. Herein, we report the case of a 43-year-old woman presenting with vaginal bleeding and pelvic mass. Histopathology of the tumor showed moderately atypical spindle cells arranged in long fascicles reminiscent of fibrosarcoma, along with immunohistochemical positivity for S100, CD34, and pan-tropomyosin receptor kinase. This prompted RNA-sequencing and the finding of a rare EML4::NTRK3 fusion. Clinical, histologic, and molecular findings are described, in addition to discussions regarding differential diagnoses and possible implications of the findings in clinical practice.

Transcriptomic investigation of the molecular mechanisms underlying resistance to the neonicotinoid thiamethoxam and the pyrethroid lambda-cyhalothrin in Euschistus heros (Hemiptera: Pentatomidae).

BACKGROUND: Laboratory-selected resistant strains of Euschistus heros to thiamethoxam (NEO) and lambda-cyhalothrin (PYR) were recently reported in Brazil. However, the mechanisms conferring resistance to these insecticides in E. heros remain unresolved. We utilized comparative transcriptome profiling and single nucleotide polymorphism (SNP) calling of susceptible and resistant strains of E. heros to investigate the molecular mechanism(s) underlying resistance. RESULTS: The E. heros transcriptome was assembled, generating 91 673 transcripts with a mean length of 720 bp and N50 of 1795 bp. Comparative gene expression analysis between the susceptible (SUS) and NEO strains identified 215 significantly differentially expressed (DE) transcripts. DE transcripts associated with the xenobiotic metabolism were all up-regulated in the NEO strain. The comparative analysis of the SUS and PYR strains identified 204 DE transcripts, including an esterase (esterase FE4), a glutathione-S-transferase, an ABC transporter (ABCC1) and aquaporins that were up-regulated in the PYR strain. We identified 9588 and 15 043 nonsynonymous SNPs in the PYR and NEO strains. One of the SNPs (D70N) detected in the NEO strain occurs in a subunit (α5) of the nAChRs, the target site of neonicotinoid insecticides. Nevertheless, this residue position in α5 is not conserved among insects. CONCLUSIONS: Neonicotinoid and pyrethroid resistance in laboratory-selected E. heros is associated with a potential metabolic resistance mechanism by the overexpression of proteins commonly involved in the three phases of xenobiotic metabolism. Together these findings provide insight into the potential basis of resistance in E. heros and will inform the development and implementation of resistance management strategies against this important pest. © 2023 Society of Chemical Industry.

Myositis Ossificans of the Breast Mimicking Breast Cancer: A Case Report.

Myositis ossificans (MO) is an uncommon tumor characterized by a rapidly growing mass following a history of local trauma. Few cases of MO affecting the breast have been reported, and some were misdiagnosed as primary osteosarcoma of the breast or metaplastic breast carcinoma. The following case report presents a patient with a growing breast lump whose core biopsy result was suspicious for breast cancer. MO was diagnosed after analysis of the mastectomy specimen. This case highlights the importance of MO as a differential diagnosis of a growing soft-tissue mass after trauma to avoid unnecessary overtreatment. Keywords: Myositis Ossificans, Osteosarcoma, Breast Cancer, Mastectomy, Heterotopic Ossification © RSNA, 2023.

Urea as a source of nitrogen and carbon leads to increased photosynthesis rates in Chlamydomonas reinhardtii under mixotrophy.

Microalgae is a potential source of bioproducts, including feedstock to biofuels. Urea has been pointed as potential N source for microalgae growth. Considering that urea metabolism releases HCO3- to the medium, we tested the hypothesis that this carbon source could improve photosynthesis and consequently growth rates of Chlamydomonas reinhardtii. In this sense, the metabolic responses of C. reinhardtii grown with ammonium and urea as nitrogen sources under mixotrophic and autotrophic conditions were investigated. Overall, the mixotrophy led to increased cell growth as well as to a higher accumulation of lipids independent of N source, followed by a decrease in photosynthesis over the growth phases. In mixotrophy, urea stimulates growth in terms of cell number and dry weight. Furthermore, higher photosynthesis was verified in late logarithmic phase compared to ammonium. Under autotrophy conditions, although cell number and biomass were reduced, there was higher production of starch independent of N source. Nonetheless, urea-based autotrophic treatments stimulated biomass production compared to ammonium-based treatment. Under mixotrophy higher input of carbon into the cell from acetate and urea optimized photosynthesis and consequently promoted cell growth. Together, these results suggest urea as alternative source of carbon, improving photosynthesis and cell growth in C. reinhardtii.

Susceptibility monitoring and comparative gene expression of susceptible and resistant strains of Spodoptera frugiperda to lambda-cyhalothrin and chlorpyrifos.

BACKGROUND: Spodoptera frugiperda (J. E. Smith) is a widespread agricultural pest with several records of resistance to different insecticides and Bt proteins, including the neurotoxic insecticides chlorpyrifos (organophosphate) and lambda-cyhalothrin (pyrethroid). Here, we (i) characterized and monitored the susceptibility of field populations of S. frugiperda to chlorpyrifos (194 populations) and lambda-cyhalothrin (197 populations) collected from major maize-growing regions of Brazil from 2003 to 2016, and (ii) compared gene expression levels of laboratory-selected, chlorpyrifos- and lambda-cyhalothrin-resistant strains to a susceptible reference strain (Sf-ss) of S. frugiperda. RESULTS: The susceptibility monitoring detected average survival ranging from 29.3% to 36.0% for chlorpyrifos, and 23.1% to 68.0% for lambda-cyhalothrin. The resistance ratio of the chlorpyrifos-resistant strain (Clo-rr) was 25.4-fold and of the lambda-cyhalothrin-resistant strain (Lam-rr) was 21.5-fold. We identified 1098 differentially expressed genes (DEGs) between Clo-rr and Sf-ss, and 303 DEGs between Lam-rr and Sf-ss. Functional analyses of the DEGs revealed the up-regulation of several detoxification enzymes, mainly cytochrome P450 belonging to CYP3 and CYP6 clans. Genes associated with regulatory processes, such as the forkhead box class O (FoxO) transcription factor were also up-regulated. Variant analysis of target-site mutations for both pesticides identified the A201S and F290V mutations in acetylcholinesterase-1, both occurring in heterozigosis in the Clo-rr S. frugiperda strain. CONCLUSION: Our data show that the overexpression of the enzymatic detoxification machinery is the main difference to explain the resistance of Clo-rr and Lam-rr strains of S. frugiperda to chlorpyrifos and lambda-cyhalothrin, although a target-site mutation also contributes to the Clo-rr resistance to chlorpyrifos. © 2023 Society of Chemical Industry.

Trends towards Lower Susceptibility of Spodoptera frugiperda (Lepidoptera: Noctuidae) to Teflubenzuron in Brazil: An Evidence for Field-Evolved Resistance.

Susceptibility monitoring to insecticides is a key component to implementing insecticide resistance management (IRM) programs. In this research, the susceptibility to teflubenzuron in Spodoptera frugiperda (J.E Smith) was monitored in more than 200 field-collected populations from major corn-growing regions of Brazil, from 2004 to 2020. Initially, we defined a diagnostic concentration of 10 µg mL-1 of teflubenzuron using a diet-overlay bioassay for monitoring the susceptibility. A variation in the susceptibility to teflubenzuron in S. frugiperda was detected among populations from different locations. We also detected a significant reduction in the susceptibility to teflubenzuron throughout time in all the populations of S. frugiperda evaluated, with larval survival at diagnostic concentration varying from values of <5% in 2004 to up 80% in 2020. Thus, this research provides evidence of field-evolved resistance of S. frugiperda to teflubenzuron and reinforces that IRM practices are urgently needed to be implemented in Brazil.

NUT carcinoma, an under-recognized malignancy: a clinicopathologic and molecular series of 6 cases showing a subset of patients with better prognosis and a rare ZNF532::NUTM1 fusion.

NUT carcinoma (NC) is a rare malignancy with aggressive clinical behavior, defined by rearrangements involving the NUTM1 gene locus. This entity is often under-recognized and its diagnosis may be challenging. In this study, we describe a subset of patients that, despite the molecularly proven diagnosis of NC, show improved outcomes. In addition, we describe one case with the novel ZNF532::NUTM1 fusion. All cases of NC diagnosed from 2013 to 2022 in our department were retrieved. FISH using dual color bring-together probes and next-generation sequencing assay were performed to characterize the fusions involving NUTM1. Among 6 patients identified, 5 were men with a median age of 35.6 years. Four patients had primary tumors in the head and neck region (2 ethmoid sinus, 1 parotid gland, and 1 lacrimal gland); 1 in the mediastinum, and another presented with a femoral bone tumor. In all cases, the initial diagnoses were not NC. The cases showed different morphological patterns, including monomorphic, rhabdoid, and pleomorphic appearances. One case showed a pseudopapillary pattern. By immunohistochemistry, all tumors showed squamous differentiation and ≥50% of neoplastic cells with nuclear positivity for NUT antibody. One case expressed WT1 (C-terminus) and other showed chromogranin positivity. Genetic study revealed a BRD4::NUTM1 fusion in all head and neck cases, BRD3::NUTM1 in mediastinum case, and ZNF532::NUTM1 fusion in the femur bone case. They were treated with surgical resection plus chemotherapy and radiotherapy. The median overall survival was 23.11 months (1.6-83.3 months) and the median disease-free survival was 14.86 months (0-54.4 months). The patients with longer overall survival were one with a lacrimal gland primary (83.3 months) and other with a parotid lesion (31.9 months). Both patients were primarily treated with complete surgical resection. Anatomic location may be directly related to the overall survival in NC cases. Resectability of the lesion is also an important factor related to survival. Pathologists should include NC in the differential diagnosis of any poorly differentiated and undifferentiated monomorphic malignancy, regardless of its anatomic location.

Pleomorphic hyalinizing angiectatic tumor of the vulva: literature review based on a rare presentation.

Pleomorphic hyalinizing angiectatic tumor (PHAT) of soft tissues is a rare, non-metastatic tumor of unknown etiology and uncertain behavior, which may recur locally. There are few reports on this condition, and due to the rarity of the disease, its lineage has not yet been fully elucidated. The present study aims to report the case of an unusual entity observed for the first time in vulval topography. A female patient, 83 years old, presented with a tumor in the vulvar region that had evolved for approximately 4 months. Magnetic resonance imaging showed an expansive perineal formation of 8.5 × 3.5 cm, and a hemivulvectomy with a flap rotation was performed. The review of the slides revealed a mesenchymal lesion without significant atypia, which was richly vascularized. In the areas of interest, the immunohistochemical (IHC) study demonstrated positivity for CD34, estrogen, and progesterone receptors; it was negative for the other tested markers. Morphological findings associated with the IHC staining panel supported the diagnosis of PHAT. The main morphological features of PHAT are clusters of ectatic vessels of different sizes that show deposits of subendothelial and intraluminal fibrin. Fusiform and pleomorphic cells randomly arranged in leaves or long fascicles intermingle these vessels. It is essential to recognize this entity and consider it among the differential diagnoses of a mesenchymal lesion, given the wide variety of entities that comprise this group of lesions.

Biomechanical responses of Nordic walking in people with Parkinson's disease.

In healthy adults, Nordic walking (NW) is known to increase the external mechanical energy fluctuations, though the external work is unaltered due to an improved pendulum-like recovery in comparison with free walking (FW). We aimed to compare mechanical, pendulum-like, and spatiotemporal parameters of gait at different speeds with and without NW poles in people with Parkinson's disease and healthy controls. The study included 11 people (aged 65.6 ± 7.0 years) with idiopathic Parkinson's disease, scoring between 1 and 1.5 on the Hoehn and Yahr scale (H&Y), and nine healthy controls (aged 70.0 ± 5.6 years). All the people were experienced Nordic walkers. Walking tests were performed at 1.8 km h-1 and 4.7 km h-1 , on eight 3D force platforms on a walkway. We found greater pendulum-like energy recovery (p < 0.05) in the Parkinson group during NW than in FW, while external mechanical work remained similar (p > 0.05). People with Parkinson's disease showed a major increase in vertical and forward energy fluctuations using poles than in healthy controls. In addition, the Parkinson group showed increased stride frequency and reduced stride length compared to controls in the NW and FW conditions. Our findings partly justify the lower walking economy in Parkinson's disease due to reduced pendulum-like mechanism at commonly used speeds. NW alters gait mechanics similarly in Parkinson group and healthy control, increasing the total mechanical work. Therefore, NW can be a compelling strategy for rehabilitation because of its potential for improving functional mobility, increasing pendulum-like mechanism in Parkinson's disease.

Pleomorphic hyalinizing angiectatic tumor of the vulva: literature review based on a rare presentation

Pleomorphic hyalinizing angiectatic tumor (PHAT) of soft tissues is a rare, non-metastatic tumor of unknown etiology and uncertain behavior, which may recur locally. There are few reports on this condition, and due to the rarity of the disease, its lineage has not yet been fully elucidated. The present study aims to report the case of an unusual entity observed for the first time in vulval topography. A female patient, 83 years old, presented with a tumor in the vulvar region that had evolved for approximately 4 months. Magnetic resonance imaging showed an expansive perineal formation of 8.5 × 3.5 cm, and a hemivulvectomy with a flap rotation was performed. The review of the slides revealed a mesenchymal lesion without significant atypia, which was richly vascularized. In the areas of interest, the immunohistochemical (IHC) study demonstrated positivity for CD34, estrogen, and progesterone receptors; it was negative for the other tested markers. Morphological findings associated with the IHC staining panel supported the diagnosis of PHAT. The main morphological features of PHAT are clusters of ectatic vessels of different sizes that show deposits of subendothelial and intraluminal fibrin. Fusiform and pleomorphic cells randomly arranged in leaves or long fascicles intermingle these vessels. It is essential to recognize this entity and consider it among the differential diagnoses of a mesenchymal lesion, given the wide variety of entities that comprise this group of lesions.

Neoadjuvant hypofractionated radiotherapy and chemotherapy for extremity soft tissue sarcomas: Safety, feasibility, and early oncologic outcomes of a phase 2 trial.

BACKGROUND AND PURPOSE: Optimal treatment of extremity soft tissue sarcomas (ESTS) is controversial. The aim of this study was to evaluate neoadjuvant chemotherapy (ChT) plus concomitant hypofractionated RT (hypo-RT) in local and distant disease relapse. Here we report safety, feasibility and early outcomes. MATERIALS AND METHODS: This was a prospective, single arm study with a goal accrual of 70 patients. Between 2015 and 2018, 18 patients with histologically confirmed nonmetastatic ESTS were assigned to receive doxorubicin and ifosfamide for three neoadjuvant cycles, concomitant with hypo-RT (25 Gy in 5 fractions) followed by surgery. The primary endpoint was disease-free survival (DFS). Secondary outcomes were pathologic response, wound complications (WC), and morbidity rates. RESULTS: Median follow-up was 29 months. At last follow-up, 13/18 patients were alive without evidence of local or systemic disease (DFS 72%), 1 had died due to metastatic disease, and 3 were alive with distant metastasis. One patient presented with local relapse within the irradiated field. Mean DFS time was 48.6 months (95% CI: 37.3-59.9). Six patients (33%) had no residual viable tumor detected in pathologic specimens (3 of these myxoid liposarcomas). There was a significant difference in WC among patients with acute RT skin toxicity. Six patients (33%) developed major WC. No grade 3 or 4 ChT adverse events were reported. CONCLUSION: Despite the limited sample size, these early outcomes demonstrate that this treatment regimen is feasible and well tolerated with high rates of limb preservation, local control, and pathologic complete response, supporting further investigation in a multi-institutional setting. TRIAL REGISTRATION: ClinicalTrials.gov NCT02812654; https://clinicaltrials.gov/ct2/show/NCT02812654.

Third-degree atrioventricular block and pacemaker implantation in a man with severe hemophilia A: A case report.

Hemophilia A (HA) is a rare bleeding disorder characterized by reduced factor VIII (FVIII) activity and consequently spontaneous bleeding. Since the introduction of prophylaxis with safer FVIII concentrates, people with HA are ageing. Interestingly, they are developing cardiovascular diseases as their non-hemophilia counterparts. We describe a 48-year-old patient with severe HA who presented a third-degree atrioventricular block. A DDDR pacemaker was implanted under supervision of the Hematology Clinics. There were no adverse events during the procedure. The procedure was safe, and it should be performed under the supervision of a hemophilia expert.

Adaptive responses of Kluyveromyces marxianus CCT 7735 to 2-phenylethanol stress: Alterations in membrane fatty-acid composition, ergosterol content, exopolysaccharide production and reduction in reactive oxygen species.

2-phenylethanol (2-PE) is a higher aromatic alcohol with a rose-like aroma used in the cosmetic and food industries as a flavoring and displays potential for application as an antifungal. Biotechnological production of 2-PE from yeast is an interesting alternative due to the non-use of toxic compounds and the generation of few by-products. Kluyveromyces marxianus CCT 7735 is a thermotolerant strain capable of producing high 2-PE titers from L-Phenylalanine; however, like other yeast species, its growth has been strongly inhibited by this alcohol. Herein, we aimed to evaluate the effect of 2-PE on cell growth, cell viability, membrane permeability, glucose uptake, metabolism, and morphology in K. marxianus CCT 7735, as well as its adaptive responses. The stress condition was imposed after 4 h of cultivation by adding 3.0 g.L-1 of 2-PE in exponential growing cells. 2-PE stress impaired yeast growth, glucose uptake, fermentative metabolism, membrane permeability, and cell viability. Moreover, the stress condition provoked changes in both morphology and surface roughness. The reactive oxygen species (ROS) increased immediately on exposure to 2-PE. Changes in membrane fatty-acid composition, ergosterol content, exopolysaccharides production, and reduction of the ROS levels appear to be the result of adaptive responses in K. marxianus. Our results provided insights into a better understanding of the effects of 2-PE on K. marxianus and its adaptive responses.

Desmoplastic Small Round Cell Tumor: A Review of Main Molecular Abnormalities and Emerging Therapy.

Desmoplastic small round cell tumor (DSRCT) is an extremely rare, aggressive sarcoma affecting adolescents and young adults with male predominance. Generally, it originates from the serosal surface of the abdominal cavity. The hallmark characteristic of DSRCT is the EWSR1-WT1 gene fusion. This translocation up-regulates the expression of PDGFRα, VEGF and other proteins related to tumor and vascular cell proliferation. Current management of DSRCT includes a combination of chemotherapy, radiation and aggressive cytoreductive surgery plus intra-peritoneal hyperthermic chemotherapy (HIPEC). Despite advances in multimodal therapy, outcomes remain poor since the majority of patients present disease recurrence and die within three years. The dismal survival makes DSRCT an orphan disease with an urgent need for new drugs. The treatment of advanced and recurrent disease with tyrosine kinase inhibitors, such as pazopanib, sunitinib, and mTOR inhibitors was evaluated by small trials. Recent studies using comprehensive molecular profiling of DSRCT identified potential therapeutic targets. In this review, we aim to describe the current studies conducted to better understand DSRCT biology and to explore the new therapeutic strategies under investigation in preclinical models and in early phase clinical trials.

Leaf application of 24-epibrassinolide mitigates cadmium toxicity in young Eucalyptus urophylla plants by modulating leaf anatomy and gas exchange.

Cadmium (Cd2+) soil pollution is a global environmental problem caused by the high toxicity of Cd. 24-Epibrassinolide (EBR) is a biodegradable plant steroid involved in response modulation to biotic and abiotic stresses. The aim of this study was to evaluate if the leaf-application of EBR improves the gas exchange and possible repercussions on leaf anatomy in young Eucalyptus urophylla plants exposed to Cd toxicity. The experiment involved six treatments, which included three Cd concentrations (0, 450, and 900 µM) and two EBR concentrations (0 and 100 nM, described as - EBR and + EBR, respectively). Plants exposed to Cd toxicity suffered decreases in leaf anatomical and gas exchange parameters. However, the plants treated with EBR + 900 µM Cd showed an increase of 46%, 40%, and 54% in the net photosynthetic rate, water-use efficiency, and instantaneous carboxylation efficiency, respectively. The EBR application-induced improvements in gas exchange parameters, causing beneficial effects on the photosynthetic apparatus, mainly the effective quantum yield of photosystem II (PSII) photochemistry and electron transport rate. Furthermore, this steroid mitigated the effect of Cd toxicity on leaf anatomical variables, more specifically palisade and spongy parenchyma, which are intrinsically related to stomatal density, and stimulated the net photosynthetic rate of plants.