Stage L1 Research Articles

N9: Pilot study of novel shortened induction for high-risk neuroblastoma.

10052 Background: N9 induction for high-risk neuroblastoma was designed to limit non-hematological toxicity (especially ototoxicity) and to prevent relapse in the central nervous system (CNS). N9 builds on 2 novel regimens: high-dose cyclophosphamide-topotecan-vincristine (CTV) and ifosfamide-carboplatin-etoposide (ICE), each with good penetration across the blood-brain barrier. Methods: N9 is a pilot study (Clinicaltrials.gov.NCT04947501) to assess feasibility and safety. Secondary/exploratory objectives include response, collection of peripheral blood stem cells (PBSCs; ≥5x106/kg CD34(+) cells sufficient for ≥2 rescues), tumor resection, and assessment of central nervous system relapse. Early stopping rules centered on excessive delay in timing of chemotherapy. Eligibility criteria included age >1-to-<13 years; 1 prior chemotherapy cycle was allowed. CTCAE Version 5.0 and International Neuroblastoma Response Criteria are used.N9 comprises 4 cycles of chemotherapy. Cycles start after absolute neutrophil count is >500/μL, platelets >100,000/μL, and non-hematologic toxicities grade ≤2. Intervals of 21-28 days between start of cycles are foreseen. PBSC collection and surgery follow >3 cycles. Cycles #1 and #4 (CTV): cyclophosphamide 70mg/kg/day, days 1-2, topotecan 2mg/m2/day, days 1-4, and vincristine 0.067mg/kg, day 1. Cycle #2 (ICE): ifosfamide 1500mg/m2/day, days 1-5, carboplatin 400mg/m2/day, days 1-2, and etoposide 100mg/m2/day, days 1-5. Cycle #3: cyclophosphamide (as in CTV) and 72-hr infusions of doxorubicin 75mg/m2 and vincristine 0.067mg/kg. Results: The target number of 15 patients were enrolled: 10/2021-9/2023; age 1.5–9.8 (median 3.3) years; 14 stage M, 1 stage L2; and 10 post-1 cycle of other chemotherapy. They completed N9 without undue toxicity: all cycles started on time, organ functions remained intact, 11/11 patients tested had no ototoxicity, and acute toxicities were typical for myelosuppression (including uncomplicated fever/neutropenia). Responses were complete (n=6), partial (n=5), and stable disease (n=4). The target number of PBSCs was collected in 12 patients (9-75, median 14 x106/kg CD34(+) cells), 4x106/kg in 2 patients, and pending in 1. All patients had gross total resections of the primary tumor. Post-N9, patients did not undergo transplant, but proceeded to immunotherapy (naxitamab) or chemoimmunotherapy (naxitamab+irinotecan-temozolomide). Of 9 with residual disease post-N9, 7 achieved CR (median of 5.6 months from study entry) and 1 achieved metabolic CR (16 months), though 3 subsequently relapsed (no CNS). Conclusions: N9 shows promise for reducing chemotherapy and long-term toxicity. Less chemotherapy without compromising survival is a realistic goal by virtue of the advances with anti-GD2 monoclonal antibodies which, in combination with GM-CSF+low-dose chemotherapy, are highly effective against chemo-resistant disease in bone/bone marrow. Clinical trial information: NCT04947501 .

APPLICATION OF THE INTERNATIONAL NEUROBLASTOMA RISK GROUP STAGING SYSTEM 2009 (INRGSS 2009) IN DIAGNOSIS AND PROGNOSIS OF CHILDHOOD NEUROBLASTOMA

Background: Neuroblastoma is the most common extracranial solid tumor of childhood. INRGSS 2009 divides patients into stages L1, L2, M, Ms, from which, in combination with histopathology, molecular biology, each patient will be divided into risk groups from which to guide prognosis and suitable treatment. Objectives: Describe the clinical and laboratory features of neuroblastoma in children; distribution of pediatric neuroblastoma stages according to the INRGSS 2009 classification and its value in predicting survival outcomes. Methods: This was a cross-sectional study combined with retrospective and prospective. Patients were diagnosed with neuroblastoma and classified according to INRGSS 2009 at the Pediatric Center, Hue Central Hospital. Results: The age at the time of diagnosis ranged from 1 day old to 9 years old, with a mean of 35.4 ± 25.5 months. Regarding primary tumor, 73.5% of cases were found in the abdomen, the mediastinum region (22.4%), an undetermined region (4.1%). Systemic symptoms, local symptoms and symptoms caused by metastasis were frequently reported. At the time of diagnosis, 73.5% of the patients had distant metastases (stage M or MS). The EFS of the whole group at 12 and 24 months old were 86.3 % and 64.7% respectively. The OS was 77.9% at 24 months old. Most of these patients had a progressive disease: 6.1% were in stage L1; 20.4% were in stage L2; 69.4% were in stage M; and 4.1% were in stage MS. Estimated mean of event-free survival for L1 was 38.7 ± 3.5 months, L2 was 40.5 ± 4.2 months and M was 28.0 ± 2.5 months (p < 0.05). Conclusions: Most neuroblastoma tumors are abdominal and have distant metastases. LDH and ferritin elevations are prevalent. Neuroblastoma outcomes can be predicted using the INRGSS 2009 classification.

Abstract 3662: Cell-free DNA dynamics following neuroblastoma resection

Abstract Introduction: Prior studies have demonstrated the utility of circulating tumor DNA (ctDNA) as a biomarker in the postoperative period for oncologic risk stratification. However, its utility may be limited in the early postoperative period by surgical trauma causing an increase in total cell-free DNA (cfDNA) due to tissue destruction, inflammation, and wound healing. No studies have identified the optimal timing of postoperative sample collection in pediatric patients undergoing resection of high-risk neuroblastoma. The purpose of this study is to assess the postoperative dynamics of cfDNA following neuroblastoma resection to determine optimal timing for ctDNA investigations. Methods: An institutional database of banked cell-free DNA samples was retrospectively queried for patients with a diagnosis of neuroblastoma who had blood samples collected prior to resection and postoperative samples collected within 2 months of the operation. Plasma was collected from whole blood samples. cfDNA was extracted from plasma samples using the QIAGEN QIAsymphony SP system, and samples were quantified using the Advanced Analytical Fragment Analyzer Automated CE System with the High Sensitivity Genomic DNA Analysis Kit. Results: Seventeen patients were identified undergoing 18 total operations and 37 total blood draws yielding cfDNA. There were 12 male and 5 female patients. Median age at operation was 5.5 years (range 1.4y to 20.5y). Two patients were INRG stage L2, and 15 patients were stage M. Operative indications were primary tumor resection in 10 patients and resection of recurrence/progression in 8 patients. Three postoperative samples were drawn within 10 days of the operation, and 16 samples were collected between postoperative weeks 4 and 6. There was a statistically significant difference (p=0.029) in cfDNA yield between each timepoint sampled: preoperative (median 6ng/mL, range 2-30ng/mL), 0-10 days postop (median 34ng/mL, range 15 - 85 ng/mL), and 4-6 weeks postop (median 6ng/mL, range 2 - 17ng/mL). Pairing the pre- and post-operative samples for each patient, those drawn within 10 days had a median increased yield of 180% (range +67% to +652%) while the samples collected at 4-6 weeks had a median decreased yield of 23% (range -55% to +491%; p = 0.047). Of note, there were three outliers in the 4-6 week group who had increases in cfDNA yield of >200%. On further review of these charts, each patient was being treated for obstructive uropathy at the time the postoperative sample was collected. Conclusion: Plasma cfDNA yield increases in the early postoperative period before dropping by postoperative week 6. Tumor resection may be associated with an overall decrease in total cfDNA. Postoperative renal pathology may cause a persistent elevation in cfDNA. Further studies are necessary to clarify the role of ctDNA in the management of patients with neuroblastoma. Citation Format: Joshua N. Honeyman, Andrew Chi, Shakeel Modak, Fiorella Iglesias Cardenas, Michael P. La Quaglia, Neerav N. Shukla, J. Ted Gerstle. Cell-free DNA dynamics following neuroblastoma resection [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3662.

Toxicity of Three Optical Brighteners: Potential Pharmacological Targets and Effects on Caenorhabditis elegans.

Optical brighteners (OBs) have become an integral part of our daily lives and culture, with a growing number of applications in various fields. Most industrially produced OBs are derived from stilbene, which has been found in environmental matrices. The main objectives for this work are as follows: first, to identify protein targets for DAST, FB-28, and FB-71, and second, to assess their effects in some behaviors physiologic of Caenorhabditis elegans. To achieve the first objective, each OB was tested against a total of 844 human proteins through molecular docking using AutoDock Vina, and affinities were employed as the main criteria to identify potential target proteins for the OB. Molecular dynamics simulations took and validated the best 25 docking results from two protein databases. The highest affinity was obtained for the Hsp70-1/DAST, CD40 ligand/FB-71, and CD40 ligand/FB-28 complexes. The possible toxic effects that OBs could cause were evaluated using the nematode C. elegans. The lethality, body length, locomotion, and reproduction were investigated in larval stage L1 or L4 of the wild-type strain N2. In addition, transgenic green fluorescent protein (GFP) strains were employed to estimate changes in relative gene expression. The effects on the inhibition of growth, locomotion, and reproduction of C. elegans nematodes exposed to DAST, FB-71, and FB-28 OBs were more noticeable with respect to lethality. Moreover, an interesting aspect in OB was increased the expression of gpx-4 and sod-4 genes associated with oxidative stress indicating a toxic response related to the generation of reactive oxygen species (ROS). In all cases, a clear concentration-response relationship was observed. It is of special attention that the use of OBs is increasing, and their different sources, such as detergents, textiles, plastics, and paper products, must also be investigated to characterize the primary emissions of OBs to the environment and to develop an adequate regulatory framework.

Contribution of differential alteration in oxidative stress and anti-oxidation related molecular signals to toxicity difference between atrazine and its main metabolites in nematodes

As a widely used herbicide, atrazine and its two main metabolites of deethylatrazine (DEA) and deisopropylatrazine (DIA) pose an exposure risk for both human beings and animals in the environment. In this study, Caenorhabditis elegans was selected as an in vivo model to compare the toxicity between atrazine and its main metabolites. Upon exposure from the larval stage L1 to adult day 3, both DEA and DIA showed less toxicity on locomotion and reproduction compared with atrazine at concentration of 0.001, 0.01 0.1 and 1 mg/L for parental generation. In addition, exposure to DEA and DIA at concentration of 0.1 mg/L also induced less transgenerational toxicity on locomotion than exposure to atrazine for both parental generation and offspring of F1–F4. Accordingly, exposure to DEA and DIA caused less ROS production and alteration in the expression of some genes (mev-1, gas-1, and clk-1) governing oxidative stress compared to atrazine. Meanwhile, DEA and DIA lead to less increase in expression of superoxide dismutase genes (sod-2 and sod-3) and SOD-3::GFP than atrazine. Moreover, atrazine and its two main metabolites differentially activated the daf-16 encoding FOXO transcriptional factor in insulin signaling pathway during the control of downstream target of SOD-3. Overall, our results highlighted the important role of oxidative stress and anti-oxidation related molecular signals in mediating toxicity of atrazine, DEA and DIA, which provided a novel explanation for the different toxicity between atrazine and its main metabolites.

Ganglioneuroblastoma intermixed: Clinicopathological implications of diagnosis at presentation and genomic correlations.

Ganglioneuroblastoma intermixed (GNBI) is classified as "favorable" histology by International Neuroblastoma Pathology Classification system. However, the International Neuroblastoma Risk Group (INRG) stratifies patients using wider clinicopathological and cytogenetic/molecular parameters. While the diagnosis of GNBI is typically made on resected tumor, it may sometimes be rendered on initial biopsy. We studied GNBI noted at diagnosis to evaluate its correlation with INRG staging and other clinicopathological and molecular features. In this retrospective study, clinical, radiological, pathological, cytogenetic, and molecular information from patients with GNBI at diagnosis seen between 1995 and 2021 was analyzed. INRG staging was performed. Of the 15,827 neuroblastoma specimens, GNBI was noted in 237 patients. Of these, 53 had the initial pathological diagnosis of GNBI; median follow-up 3.5 (range: 0.2-14) years. Disease was locoregional in 41 (77%, 16 stage L1 and 25 L2); none relapsed. Twelve (23%) had metastatic disease at presentation; six (50%) relapsed, and two died of disease. MYCN was amplified in two metastatic tumors. Six of 31 (19%) tumors tested had recurrent cytogenetic abnormalities and nonrecurrent somatic gene mutations in 10/23 (43%). The presence of any adverse molecular/cytogenetic findings was associated with metastatic disease (p<.05). For patients with localized GNBI undergoing both biopsy and resection, GNBI was diagnosed in both in 17/19 (90%). Localized GNBI at diagnosis has excellent long-term clinical outcome even without cytotoxic therapy. For localized GNBI, a biopsy sample is adequate to make the diagnosis. When associated with metastasis at diagnosis, prognosis is poorer, possibly due to associated adverse biological features.

Association of image-defined risk factors with clinical features, tumor biology, and outcomes in neuroblastoma: a single-center retrospective study.

Image-defined risk factors (IDRF) in neuroblastoma have been developed to predict tumor resectability and surgical complications; however, the potential prognostic value of IDRF in neuroblastoma has been variably reported. Previous studies did not report the IDRF status separately from the International Neuroblastoma Risk Group (INRG) stage. Moreover, the association between IDRF and clinical and pathological factors has not been discussed further. In this retrospective study, we investigated the clinical and biological features of neuroblastoma at different INRG stages based on IDRF. Event-free survival (EFS) and overall survival (OS) related to the INRG stage were analyzed using log-rank tests, and the prognostic value of the IDRF number and type was also evaluated. Among 72 patients, 182 IDRF at diagnosis were found in 79.2%. The distribution of the INRG stages was 10 L1 (13.9.0%), 25 L2 (34.7%), and 37M/MS (51.4%). Patients with stage M/Ms had a larger tumor volume, a higher percentage of age ≥ 18months, elevated lactate dehydrogenase (LDH) level, elevated ferritin level, and a higher percentage of COG high-risk compared with stage L1 and L2 patients. EFS and OS were similar for stage L1 and L2 tumors but were significantly poorer for metastatic disease. However, EFS (P = 0.06) and OS (P = 0.07) were similar for IDRF-negative and positive neuroblastomas. Patients with stage M/Ms with IDRF-positive had poorer EFS (P = 0.001) and OS (P < 0.001) compared with patients in stage L2. An IDRF ≥ 4, vascular IDRF, and infiltrative IDRF of the tumor were significant indicators of poor prognosis. Conclusion: Our study indicates that increasing the INRG stages based on IDRF is associated with various unfavorable clinical features of neuroblastoma. The principal determinant of survival in neuroblastoma is the presence of metastatic disease more than IDRF alone at diagnosis. Both the number and type of IDRF have important clinical significance in the protocol planning of neuroblastoma, rather than just considering the absence or presence of IDRF. What is Known: • The International Neuroblastoma Risk Group Staging System (INRGSS) now employs image-defined risk factors (IDRFs) to stratify and stage disease. • The presence of IDRF at diagnosis are associated with higher rates of operative complications and incomplete surgical resection. What is New: • The principal determinant of survival from neuroblastoma is the presence of metastatic disease at diagnosis, more than IDRF alone. • IDRF number and type should also be considered during the diagnosis and treatment planning of neuroblastoma, rather than just considering the absence or presence of IDRF.

7,10,13,16-Docosatetraenoic acid impairs neurobehavioral development by increasing reactive oxidative species production in Caenorhabditis elegans

AimsTo investigate human breast milk (HBM) lipids that may adversely affect infant neurodevelopment. Main methodsWe performed multivariate analyses that combined lipidomics and psychologic Bayley-III scales to identify which HBM lipids are involved in regulating infant neurodevelopment. We observed a significant moderate negative correlation between 7,10,13,16-docosatetraenoic acid (omega-6, C22H36O2, the common name adrenic acid, AdA) and adaptive behavioral development. We further studied the effects of AdA on neurodevelopment by using Caenorhabditis elegans (C. elegans) as a model. Worms from larval stages L1 to L4 were supplemented with AdA at 5 nominal concentrations (0 μM [control], 0.1 μM, 1 μM, 10 μM, and 100 μM) and subjected to behavioral and mechanistic analyses. Key findingsSupplementation with AdA from larval stages L1 to L4 impaired neurobehavioral development, such as locomotive behaviors, foraging ability, chemotaxis behavior, and aggregation behavior. Furthermore, AdA upregulated the production of intracellular reactive oxygen species. AdA-induced oxidative stress blocked serotonin synthesis and serotoninergic neuron activity and inhibited expression of daf-16 and the daf-16–regulated genes mtl-1, mtl-2, sod-1, and sod-3, resulting in attenuation of the lifespan in C. elegans. SignificanceOur study reveals that AdA is a harmful HBM lipid that may have adverse effects on infant adaptive behavioral development. We believe this information may be critical for AdA administration guidance in children's health care.

Neuroblastic tumours in children: Outcome from a referral centre in India.

There is a paucity of uniform literature on the outcome of children with neuroblastic tumours from developing countries. This study aims to present the outcome in children having neuroblastic tumours. We retrospectively reviewed patients with neuroblastic tumours from January 2014 to March 2020. Data analysed were pertaining to the age, sex, presentation, site, stage, diagnostic evaluation performed, management and follow-up results, etc., The final outcomes were assessed as complete response; partial response (PR); no response (NR) and progressive disease. International Neuroblastoma Risk Group staging was used and patients were categorised on the basis of age, site and stage of tumour. Overall survival (OS) was calculated from the date of diagnosis to the date of last follow-up and event for OS was death. Forty-seven patients were included with median age of 60 months (ranges of 2-180; mean = 61.87 ± 47.56). About 72.3% (n = 34) patients were males. Out of total, 6.4% (n = 3), 53.2% (n = 25) and 38.3% (n = 18) were in stage L1, L2 and M, respectively, whereas 2.1% (n = 1) patients were in stage MS. Surgery was performed in 25 (53.19%) patients, but gross total excision was achieved in 48.90% (n = 23) patients. Onlu 10.60% (n = 5) patients were referred, whereas 14.89% (n = 7) patients reported recurrences. Mean and median follow-up time of the patients was 24.64 ± 16.04 and 18 months (range: 3-60 months). Out of total, 53.2% (n = 25) and 29.8% (n = 14) patients had complete and PRs, respectively, whereas 17% (n = 8) patients had NR. Out of the total 47 patients, 8 (17%) achieved events (deaths), whereas the rest, 39 (83%) patients, were censored. Mean event-free OS time was 50.04 months. There was a significant difference in patient deaths in recurrence and non-recurred patients (4/7, 57.1% vs. 4/40/, 10%, P = 0.011). Survival time was significantly higher in patients with stages L1-L2 as compared to Stage 4. Stage and age were found predictors of survival.

Biochemical analysis in congenital neuroblastoma.

The incidence of congenital neuroblastoma has increased in the recent years. The purpose of this study was to describe the clinical and biochemical characteristics of cases of congenital neuroblastoma diagnosed in our center. We report three cases of congenital neuroblastoma diagnosed in our hospital. In two, diagnosis was made prenatally, whereas the other case was detected in the immediate neonatal period. In the three cases, neuroblastoma was located in the abdominal region and exhibited elevated concentrations of catecholamines or their metabolites in single voided urine samples. Two tumors were classified as stage M, and one as stage L2. The N-MYC oncogen was not amplified in any of the cases studied. Histopathological analysis was favorable in the three cases. The tumor was resected in two patients. The three received chemotherapy. The measurement of catecholamines and their metabolites is essential in the diagnosis of neuroblastoma. When 24h urine cannot be collected, single voided urine can be used to calculate the index based on creatinine concentrations.

Cortisol Level and Na+/K+-ATPase Activity during Growth and Development of Juvenile Daubed Shanny Leptoclinus maculatus (Fries, 1838) in the Arctic

The dynamics of cortisol level and Na+/K+-ATPase activity that are associated with the maintenance of cell ion homeostasis was studied in skeletal muscles of juvenile circumpolar fish daubed shanny Leptoclinus maculatus (Fries, 1838), an ecologically significant representative of the ichthyofauna of the Arctic Svalbard Archipelago. It was established that the level of cortisol and Na+/K+-ATPase activity decrease during the development of daubed shanny juveniles from stage L2 pelagic juveniles to stage L5 juveniles, which are mainly demersal. The results obtained suggest that cortisol may be involved in the regulation of the activity of one of the main osmoregulatory factors, Na+/K+-ATPase. This may be important for the growth and adaptation of pelagic juveniles of daubed shanny to the demersal habitat during postembryonic development in the Arctic.

Neuroblastoma: Application of International Neuroblastoma Pathology Classification on fine needle aspiration cytology smears

Neuroblastoma (NB) is the fourth most common tumor of childhood. There is a paucity of literature on its subtyping of cytology and prognostic utility. We aimed to study the cytopathological features of NB on the aspirated material, subtype it, and assess the role of International Neuroblastoma Pathology Classification (INPC) classification on cytology smears in the preoperative prognosis of NB. Fifteen cases of NB reported on fine-needle aspiration cytology (FNAC) in the past 3 years were included. Detailed clinical, radiological, and cytological features were noted. Smears were assessed for characteristics such as cellularity, neuroblasts (cytoplasmic, nuclear details), rosettes, neuropil, Schwann cells, fibroblasts, calcification, and necrosis. Afterward, cases were categorized as undifferentiated (UD), poorly differentiated (PD), and differentiating (D) subtypes. Mitotic-karyorrhectic index (MKI) was calculated and correlated with histopathology. Follow-up was done to date. The age ranged from 19 days to 10 years with an M: F ratio of 3:1. Twelve cases were retroperitoneal, two cervical, and one mediastinal. Metastatic disease was seen in six cases, one to the cervical, four to the bone marrow, and two to the scalp. The International Neuroblastoma Risk Group (INRG) staging system was available in all cases, out of which three were in stage L1, six in stage L2, four in stage M, and two in stage Ms. On cytology, four cases were differentiating NB, five PD NB, and six UD NB. The MKI was high (>4%) in 80% of UD, intermediate (2-4%) in 100% of PD, and low (<2%) in 75% of D cases. MKI corroborated in both histology and cytology, except in one case. NB can be subtyped on cytology on the basis of characteristics of neuroblasts, presence of neutrophils, rosettes, and necrosis. UD NB has a high MKI and is associated with a poor prognosis. A preoperative comprehensive reporting of NB on cytology can be very useful in guiding appropriate chemotherapy with some increment in survival. However, larger studies are needed to validate the calculation of MKI on FNA smears.

COMPARATIVE ANALYSIS OF HYPOCHLORITE METHOD ON TRANSGENIC STRAIN BY250

Caenorhabditis elegans is an ideal model organism that has been applied to a wide range of studies into toxicology, neurobiology, molecular, and developmental biology. C.elegans has gain popularity as a model system worldwide because of its availability of genome database and similarity of the genome to human have enlarged its applications in the different areas of research and to study the molecular mechanism of various human diseases such as neurodegenerative, metabolic, and so on. The researchers were synchronized C.elegans to larval stage L1 before they experimented. These studies were propagated and describe as a powerful tool to obtain a large population of worms through a synchronization process at a specific stage, which can be used for a variety of experiments.

Safety and Feasibility of Laparoscopic Resection of Neuroblastoma Without Image-Defined Risk Factors Performed by Pediatric Surgical Trainees: A Multicenter Comparison Study.

Introduction: Laparoscopic resection has gradually been adopted for neuroblastoma patients; however, some authors reported that, due to its technically demanding procedures, this operation should be performed only by highly experienced surgeons. The aim of this study was to evaluate the safety and feasibility of laparoscopic resection of abdominal neuroblastoma by pediatric surgical trainees. Subjects and Methods: In this multicenter, retrospective study, including 18 children with abdominal neuroblastoma indicated for 19 laparoscopic resections (1 with bilateral neuroblastomas) from 1999 to 2018, the clinical data were retrospectively reviewed and compared between trainee and attending surgeons. Results: None of the cases had image-defined risk factors (IDRFs) at surgery. All patients successfully underwent complete laparoscopic resection without blood transfusion, open conversion, or intraoperative or postoperative complications. Of the 19 cases, 6 were performed by pediatric surgical trainees under the guidance of attending surgeons, and 13 were performed by attending surgeons. With comparable tumor, largest diameter, operative time, and bleeding amount were not significantly different between the two groups. In the trainee surgeon group, one local recurrence occurred at a secondary resection site in the bilateral neuroblastoma patient with Stage L2. Conclusions: Laparoscopic resection of neuroblastoma could be safe and feasible when limited to absent IDRFs at surgery by pediatric surgical trainees under the guidance of experienced attending surgeons, as well as by attending surgeons.

The use of different sublethal endpoints to monitor atrazine toxicity in nematode Caenorhabditis elegans

In this work, Caenorhabditis elegans was employed as an in vivo model to determine the toxic effects of atrazine at different concentrations. After the exposure period from the larval stage L1 to adulthood day 1, atrazine (10 mg/L) significantly decreased the body length and lifespan of nematodes. In addition, exposure to ≥0.01 mg/L atrazine remarkably increased the intestinal reactive oxygen species (ROS) levels and reduced locomotion behavior of nematodes, while exposure to ≥ 1 mg/L atrazine decreased the brood size of nematodes. Moreover, atrazine (0.001–0.1 mg/L) upregulated the expression levels of hsp-6::GFP and hsp-6/60 in nematodes, indicating the activation of mitochondrial unfolded protein response (mtUPR). On the contrary, atrazine (1–10 mg/L) downregulated the expression levels of hsp-6::GFP and hsp-6/60 in nematodes. Furthermore, mtUPR induction governed by the RNAi knockdown of atfs-1 could increase the vulnerability of nematodes against atrazine toxicity. Overall, our findings highlighted the dynamic responses of nematodes toward different concentrations of atrazine, which could be monitored using different sublethal endpoints as bioindicators.

Comparison of the effectiveness of the most used chemicals against Tuta absoluta in Tunisia

Control failures with several chemicals have prompted research into the resistance status of Tuta absoluta and the effectiveness of certain active ingredients against this pest. In this context, a comparison was made of the effectiveness of the most widely used chemicals against T. absoluta in Tunisia. The results show that the L1 larval stage of Tuta absoluta is very sensitive to all of the chemicals tested. Stage L2 is sensitive to Spinosad, indoxacarb and emactin, while Bacillus thuringiensis, martine do not cause 50% mortality. L3 stage mortality fluctuates between 20% and 80% mortality. The L4 stage is more affected by the treatment with Bacillus thuringiensis which can go to 100%. Based on these results, chemical treatments should be directed according to the dominant larval stage in the plants.

Implication of Image-Defined Risk Factors for the Extent of Surgical Resection and Clinical Outcome in Patients with Pelvic Neuroblastoma.

Pelvic neuroblastoma (NB) is a rare entity and occurs in 2 to 5% of all NBs. Surgery in the pelvic area is-even for the experienced oncological surgeon-technically challenging, as injuries of bladder and/or rectal innervation may carry lifelong consequences for the patient. Several studies have proven the impact of image-defined risk factors (IDRFs) for outcome, complications and extent of resection in NB; however, the specific role of IDRF in pelvic NB has not been investigated yet. Patient charts were retrospectively evaluated for International Staging System stage, IDRF status, MYCN amplification, and outcome parameters. Between 2003 and 2019, 277 NBs were surgically resected in the department of pediatric surgery of Dr. von Hauner Children's Hospital. Out of these, 11 patients (3.9%) had pelvic NB. Evaluation of the preoperative imaging showed two patients without IDRF (stage L1) and eight patients in stage L2. One patient had stage M according to distant metastasis. Patients without IDRF underwent complete macroscopical resections, whereas complete tumor removal was not possible without mutilation in patients with IDRF. At time point of diagnosis, only patients with IDRF had functional neurological problems. Three patients developed perioperative complications; all of them had at least one IDRF. Three patients developed local recurrence during the course of the disease, all of them had at least one IDRF. Our results indicate on a preliminary level the importance of IDRF as a prognostic tool for surgical removal of pelvic NB.

Role of image-defined risk factors in predicting surgical complications of localized neuroblastoma.

PurposeTo explore the relationship between image-defined risk factors and surgical complications of localized neuroblastoma.MethodsWe retrospectively evaluated 84 patients who met the inclusion criteria at our hospitals between June 2014 and June 2019. Patients’ clinic data were collected and the common terminology criteria for adverse events were used to categorize complications as major (grade 3–4) or minor (grade 1–2).ResultsFour (11.8%) out of 34 stage L1 patients and 15 (30.0%) out of 50 stage L2 patients had surgical complications (P < 0.05). Seventy patents underwent primary surgery, including all stage L1 patients and 36 stage L2 patients. There were no significant differences between the two groups regarding surgical complications or major surgical complications. Among stage L2 patients, 2 (5.6%) out of 36 who underwent primary surgery and 2 (14.3%) out of 14 who underwent secondary surgery had major surgical complications (P < 0.05). Complete tumor resection was achieved in 18 (50%) and 7 (50%) patients in each group (P > 0.05). The mean numbers of IDRFs were 2.06 and 4.29, respectively (P < 0.05).ConclusionsLocalized neuroblastoma patients with IDRFs have a greater surgical risk. And the number of IDRFs is not ignorable, especially in predicting major surgical complications.

Pathogistological changes in the lungs of Felis silvestris catus when infected with Aelurostrongylus abstrusus

Aelurostrongylus abstrusus is a parasite that belongs to Nematoda type, Metastrongyloidea superfamily, Angiostrongylidae family, Aelurostrongylus genus and is known as feline helminth, whose intermediate hosts are slugs and snails and reservoir hosts are mice and birds. In cats, Aelurostrongylus abstrusus parasitizes in bronchioles and alveoli, causing specific inflammatory changes in the lower respiratory tract and enables the development of such clinical signs as cough, shortness of breath as well as mucous-purulent discharge from the nasal cavity, when complicated by bacterial infections. Females of parasite lay eggs, which the larvae of L1 hatch from. Subsequently, these larvae cough with bronchial mucus and are swallowed by an animal. They pass through the digestive channel without changes and together with the faeces are released into the environment. The first stage larvae actively penetrate into the body of the slugs, fade twice and are becoming invasive. Duration of larvae development from the stage L1 to L3 depends on the biological cycle of intermediate hosts – slugs and snails. Cats are becoming infected by eating slugs or snails with invasive larvae of the third stage (L3), which are transmitted from the digestive channel of the definitive host (cat) with blood flow to the lungs. Usually, the pathogen has low pathogenicity and invasions are primarily detected after animal’s death, during pathoanatomical dissection. Acute clinical signs of the disease have not been described much up to date. The article presents the results of pathomorphological examination of the cat's lungs infected by Aelurostrongylus abstrusus. The severity of pathomorphological changes of the cat’s pulmonary tissue depends on the intensity of the invasion. When dissected, the lungs appear unevenly coloured, dark red with light and slightly denser areas, where focal parasitic nodules are clearly distinct, surrounded by pink halo and different in size but mostly round, dense consistency. Main morphological characteristic of Aelurostrongylus abstrusus infection is the presence of numerous eggs and L1 larvae in the alveoli but also in the bronchi. Helminth eggs accumulated in the alveolar lumps, surrounded by a thin shell; a severe cellular reaction from lymphocytes, macrophages and histiocytes occurred around this shell. An inflammatory reaction resulted in the thickening of the interstitial tissue with a strong vascular response, characterized by fraying and edema of the vessel walls. The results of pathomorphological researches have shown that the main changes were localized in the lungs and bronchial tubes with diffuse damage of the pulmonary tissue parenchyma and development of croupous pneumonia.

Larvicidal activity of Bacillus Thuringiensis Kurstaki against Tuta Absoluta (Lepidoptera: Gelechiidae)

A commercial formulation of Bacillus thurengiensis var Kurstaki (BTK) (IAB Bt) has been shown to be effective in controlling Tutaabsoluta Zeller larvae under laboratory conditions. Five concentrations are tested ((200, .350;500;650et800ppm)by ingestion, on the various larval stages (L1, L2, L3, L4 and L5). The larval mortality caused by BTK swelledwith increasing concentrations. The high concentrations of 650 and 800 ppm give 100% mortality rates for the early stages L1, L2, L3 and L4. Theacute toxicity (LC50) of BTK calculated for all larvae is 517 ppm. The lethal efficacy of BTK is noted 02 days, after the ingestion treatment, for the young stages (L1 and L2) and 04 at 6 days for the older larvae (L3 and L4) with a LT50, all larval stages combined equal to 2.6 days.