SAMHD1 acts at stalled replication forks to prevent interferon induction.

SAMHD1 was previously characterized as a dNTPase that protects cells from viral infections. Mutations in SAMHD1 are implicated in cancer development and in a severe congenital inflammatory disease known as Aicardi-Goutières syndrome. The mechanism by which SAMHD1 protects against cancer and chronic inflammation is unknown. Here we show that SAMHD1 promotes degradation of nascent DNA at stalled replication forks in human cell lines by stimulating the exonuclease activity of MRE11. This function activates the ATR-CHK1 checkpoint and allows the forks to restart replication. In SAMHD1-depleted cells, single-stranded DNA fragments are released from stalled forks and accumulate in the cytosol, where they activate the cGAS-STING pathway to induce expression of pro-inflammatory type I interferons. SAMHD1 is thus an important player in the replication stress response, which prevents chronic inflammation by limiting the release of single-stranded DNA from stalled replication forks.

Telemedicine Beyond Borders: A Mixed-Methods Approach to Understanding Cross-National Usage Intention.

Introduction: Drawing on the Unified Theory of Acceptance and Use of Technology 2 and the Diffusion of Innovation Theory, this article investigates the adoption of telemedicine services from a patient perspective in Germany, Spain, and the United States using a mixed-methods approach. Digital health technologies have the potential to improve access to care and to alleviate the burden on traditional health care systems and are becoming more integrated into everyday medicine. Therefore, understanding the factors that impact patients' intentions to use telemedicine is crucial to ensure successful development. Methods: Based on 1,200 surveys collected in Germany, Spain, and the United States, structural equation modeling (IBM SPSS Amos 24) is employed to test the hypotheses. The article also explores how age and gender moderate the proposed relationships. Results: Seven out of the 10 hypotheses (performance expectancy, hedonic motivation, habit, relative advantage, and perceived security) are found to be positive, direct, and statistically significant. Furthermore, findings suggest stronger effects for telemedicine usage intention for younger female users than their male counterparts. Discussion: With digital health technologies becoming more prevalent, the outcomes of this study can endorse the development of effective strategies to promote the adoption of telemedicine, ultimately improving access to care and contributing to the advancement of and modern health care.

High Sensitivity Near-Infrared Imaging of Fluorescent Nanosensors.

Biochemical processes are fast and occur on small-length scales, which makes them difficult to measure. Optical nanosensors based on single-wall carbon nanotubes (SWCNTs) are able to capture such dynamics. They fluoresce in the near-infrared (NIR, 850-1700 nm) tissue transparency window and the emission wavelength depends on their chirality. However, NIR imaging requires specialized indium gallium arsenide (InGaAs) cameras with a typically low resolution because the quantum yield of normal Si-based cameras rapidly decreases in the NIR. Here, an efficient one-step phase separation approach to isolate monochiral (6,4)-SWCNTs (880 nm emission) from mixed SWCNT samples is developed. It enables imaging them in the NIR with high-resolution standard Si-based cameras (>50× more pixels). (6,4)-SWCNTs modified with (GT)10 -ssDNA become highly sensitive to the important neurotransmitter dopamine. These sensors are 1.7× brighter and 7.5× more sensitive and allow fast imaging (<50 ms). They enable high-resolution imaging of dopamine release from cells. Thus, the assembly of biosensors from (6,4)-SWCNTs combines the advantages of nanosensors working in the NIR with the sensitivity of (Si-based) cameras and enables broad usage of these nanomaterials.

Interdependencies of cellular and humoral immune responses in heterologous and homologous SARS-CoV-2 vaccination.

BACKGROUND: Homologous and heterologous SARS-CoV-2 vaccinations yield different spike protein-directed humoral and cellular immune responses. This study aimed to explore their currently unknown interdependencies. METHODS: COV-ADAPT is a prospective, observational cohort study of 417 healthcare workers who received vaccination with homologous ChAdOx1 nCoV-19, homologous BNT162b2 or with heterologous ChAdOx1 nCoV-19/BNT162b2. We assessed humoral (anti-spike-RBD-IgG, neutralizing antibodies, and avidity) and cellular (spike-induced T-cell interferon-γ release) immune responses in blood samples up to 2 weeks before (T1) and 2-12 weeks following secondary immunization (T2). RESULTS: Initial vaccination with ChAdOx1 nCoV-19 resulted in lower anti-spike-RBD-IgG compared with BNT162b2 (70 ± 114 vs. 226 ± 279 BAU/ml, p < .01) at T1. Booster vaccination with BNT162b2 proved superior to ChAdOx1 nCoV-19 at T2 (anti-spike-RBD-IgG: ChAdOx1 nCoV-19/BNT162b2 2387 ± 1627 and homologous BNT162b2 3202 ± 2184 vs. homologous ChAdOx1 nCoV-19 413 ± 461 BAU/ml, both p < .001; spike-induced T-cell interferon-γ release: ChAdOx1 nCoV-19/BNT162b2 5069 ± 6733 and homologous BNT162b2 4880 ± 7570 vs. homologous ChAdOx1 nCoV-19 1152 ± 2243 mIU/ml, both p < .001). No significant differences were detected between BNT162b2-boostered groups at T2. For ChAdOx1 nCoV-19, no booster effect on T-cell activation could be observed. We found associations between anti-spike-RBD-IgG levels (ChAdOx1 nCoV-19/BNT162b2 and homologous BNT162b2) and T-cell responses (homologous ChAdOx1 nCoV-19 and ChAdOx1 nCoV-19/BNT162b2) from T1 to T2. Additionally, anti-spike-RBD-IgG and T-cell response were linked at both time points (all groups combined). All regimes yielded neutralizing antibodies and increased antibody avidity at T2. CONCLUSIONS: Interdependencies between humoral and cellular immune responses differ between common SARS-CoV-2 vaccination regimes. T-cell activation is unlikely to compensate for poor humoral responses.

Human-Robot Collaboration in Industrial Automation: Sensors and Algorithms.

Technology is changing the manufacturing world [...].

Transvascular therapy of Hepatocellular Carcinoma (HCC), status and developments.

Hepatocellular carcinoma (HCC) is the fifth most common cancer worldwide. Only 30-40% of patients diagnosed with HCC are candidates for curative treatment options. The remaining majority of patients undergo local, regional or systemic palliative therapies. Transvascular therapy of HCC takes advantage of the fact that hypervascularized HCCs receive their main perfusion from the hepatic artery. In this context transvascular therapy describes different therapies: bland embolization (transarterial embolization, TAE), cTACE (conventional transarterial chemoembolization), DEB-TACE (TACE with drug-eluting beads, DEB) and SIRT (selective internal radiation therapy, radioembolization). cTACE is the most common type of transvascular treatment and represents a combination of the intra-arterial use of a chemotherapeutic agent and embolization. There is no standardized regimen for cTACE. It remains unclear whether the intra-arterial application of a chemotherapeutic agent is definitely required, because bland embolization alone using very small spherical particles shows tumor necrosis comparable to cTACE. For DEB-TACE microparticles loaded with a chemotherapeutic drug combine the advantages of cTACE and bland embolization.

Evaluation of the plasmatic and parenchymal elution kinetics of two different irinotecan-loaded drug-eluting embolics in a pig model.

PURPOSE: To evaluate and compare irinotecan elution kinetics of two drug-eluting embolic agents in a porcine model. MATERIALS AND METHODS: Embolization of the left liver lobe was performed in 16 domestic pigs, with groups of two receiving 1 mL of DC Bead M1 (70-150 µm) or Embozene TANDEM (75 µm) loaded with 50 mg irinotecan. Irinotecan plasma levels were measured at 0, 10, 20, 30, 60, 120, 180, and 240 minutes after completed embolization and at the time of euthanasia (24 h, 48 h, 72 h, or 7 d). Liver tissue samples were taken to measure irinotecan tissue concentrations. RESULTS: The highest irinotecan plasma concentrations of both embolic agents were measured 10 and 20 minutes after embolization, and concentrations were significantly higher for DC Bead M1 versus Embozene TANDEM (P = .0019 and P = .0379, respectively). At 48 hours and later follow-up, no irinotecan was measurable in the plasma. For both embolic agents, the highest irinotecan tissue concentration was found after 24 hours and decreased in a time-dependent manner at later follow-up intervals. Additionally, SN-38 tissue levels for both agents were therapeutic at 24 hours, with therapeutic levels of SN-38 at 48 hours in one liver embolized with TANDEM particles. Histopathologic analysis revealed ischemic, inflammatory, and fibrotic tissue reactions. CONCLUSIONS: Irinotecan is measurable in plasma and hepatic tissue after liver embolization with both types of irinotecan-eluting embolic agents. DC Bead M1 shows early burst elution kinetics, whereas Embozene TANDEM has a lower and slower release profile. The initial burst is significantly greater after embolization with DC Bead M1 than with Embozene TANDEM.

Specific CT 3D rendering of the treatment zone after Irreversible Electroporation (IRE) in a pig liver model: the "Chebyshev Center Concept" to define the maximum treatable tumor size.

BACKGROUND: Size and shape of the treatment zone after Irreversible electroporation (IRE) can be difficult to depict due to the use of multiple applicators with complex spatial configuration. Exact geometrical definition of the treatment zone, however, is mandatory for acute treatment control since incomplete tumor coverage results in limited oncological outcome. In this study, the "Chebyshev Center Concept" was introduced for CT 3d rendering to assess size and position of the maximum treatable tumor at a specific safety margin. METHODS: In seven pig livers, three different IRE protocols were applied to create treatment zones of different size and shape: Protocol 1 (n = 5 IREs), Protocol 2 (n = 5 IREs), and Protocol 3 (n = 5 IREs). Contrast-enhanced CT was used to assess the treatment zones. Technique A consisted of a semi-automated software prototype for CT 3d rendering with the "Chebyshev Center Concept" implemented (the "Chebyshev Center" is the center of the largest inscribed sphere within the treatment zone) with automated definition of parameters for size, shape and position. Technique B consisted of standard CT 3d analysis with manual definition of the same parameters but position. RESULTS: For Protocol 1 and 2, short diameter of the treatment zone and diameter of the largest inscribed sphere within the treatment zone were not significantly different between Technique A and B. For Protocol 3, short diameter of the treatment zone and diameter of the largest inscribed sphere within the treatment zone were significantly smaller for Technique A compared with Technique B (41.1 ± 13.1 mm versus 53.8 ± 1.1 mm and 39.0 ± 8.4 mm versus 53.8 ± 1.1 mm; p < 0.05 and p < 0.01). For Protocol 1, 2 and 3, sphericity of the treatment zone was significantly larger for Technique A compared with B. CONCLUSIONS: Regarding size and shape of the treatment zone after IRE, CT 3d rendering with the "Chebyshev Center Concept" implemented provides significantly different results compared with standard CT 3d analysis. Since the latter overestimates the size of the treatment zone, the "Chebyshev Center Concept" could be used for a more objective acute treatment control.

Co-simulation of neuromuscular dynamics and knee mechanics during human walking.

This study introduces a framework for co-simulating neuromuscular dynamics and knee joint mechanics during gait. A knee model was developed that included 17 ligament bundles and a representation of the distributed contact between a femoral component and tibial insert surface. The knee was incorporated into a forward dynamics musculoskeletal model of the lower extremity. A computed muscle control algorithm was then used to modulate the muscle excitations to drive the model to closely track measured hip, knee, and ankle angle trajectories of a subject walking overground with an instrumented knee replacement. The resulting simulations predicted the muscle forces, ligament forces, secondary knee kinematics, and tibiofemoral contact loads. Model-predicted tibiofemoral contact forces were of comparable magnitudes to experimental measurements, with peak medial (1.95 body weight (BW)) and total (2.76 BW) contact forces within 4-17% of measured values. Average root-mean-square errors over a gait cycle were 0.26, 0.42, and 0.51 BW for the medial, lateral, and total contact forces, respectively. The model was subsequently used to predict variations in joint contact pressure that could arise by altering the frontal plane joint alignment. Small variations (±2 deg) in the alignment of the femoral component and tibial insert did not substantially affect the location of contact pressure, but did alter the medio-lateral distribution of load and internal tibia rotation in swing. Thus, the computational framework can be used to virtually assess the coupled influence of both physiological and design factors on in vivo joint mechanics and performance.

Emergency embolization for the treatment of acute hemorrhage from intercostal arteries.

To evaluate embolotherapy for the emergency management of acute bleeding from intercostal arteries. Between October 2003 and August 2012, 19 consecutive patients with hemorrhage from intercostal arteries were scheduled for emergency embolization. The primary study endpoints were technical and clinical success, which were defined as angiographic cessation of bleeding, and cessation of clinical signs of hemorrhage. The secondary study endpoints were periprocedural complications and 30-day mortality rate. In most patients (74 %), hemorrhage was caused by iatrogenic procedures with subsequent intercostal artery laceration. One of the patients was treated twice for recurrent hemothorax caused by a new intercostal artery pseudoaneurysm 7.5 years after the initial procedure. Thus, 20 procedures were performed in these 19 patients. Overall technical success was 85 %. In six patients, no embolization of the "backdoor" was feasible, and in two of these patients additional embolization of other intercostal arteries was necessary to prevent hemorrhage via collateral vessels. Clinical signs of hemorrhage ceased after embolotherapy in 16 of 20 procedures (clinical success 80 %). The mean follow-up was 358.7 ± 637.1 days. One minor procedure-related complication occurred. The 30-day mortality rate was 21 %, however, this was unrelated to intercostal artery hemorrhage. Embolotherapy is an effective emergency therapy for patients with acute hemorrhage from intercostal arteries. Especially if embolization of the backdoor is not feasible, collateral supply via other intercostal arteries should be either ruled out or embolized to prevent ongoing hemorrhage. Despite successful embolotherapy, a majority of patients underwent surgery during follow-up to remove the symptomatic hematoma.

Novel variants in CSF1R associated with adult-onset leukoencephalopathy with axonal spheroids and pigmented glia (ALSP).

The CSF1R gene, located on chromosome 5, encodes a 108 kDa protein and plays a critical role in regulating myeloid cell function. Mutations in CSF1R have been identified as a cause of a rare white matter disease called adult-onset leukoencephalopathy with axonal spheroids and pigmented glia (ALSP, also known as CSF1R-related leukoencephalopathy), characterized by progressive neurological dysfunction. This study aimed to broaden the genetic basis of ALSP by identifying novel CSF1R variants in patients with characteristic clinical and imaging features of ALSP. Genetic analysis was performed through whole-exome sequencing or panel analysis for leukodystrophy genes. Variant annotation and classification were conducted using computational tools, and the identified variants were categorized following the recommendations of the American College of Medical Genetics and Genomics (ACMG). To assess the evolutionary conservation of the novel variants within the CSF1R protein, amino acid sequences were compared across different species. The study identified six previously unreported CSF1R variants (c.2384G>T, c.2133_2919del, c.1837G>A, c.2304C>A, c.2517G>T, c.2642C>T) in seven patients with ALSP, contributing to the expanding knowledge of the genetic diversity underlying this rare disease. The analysis revealed considerable genetic and clinical heterogeneity among these patients. The findings emphasize the need for a comprehensive understanding of the genetic basis of rare diseases like ALSP and underscored the importance of genetic testing, even in cases with no family history of the disease. The study's contribution to the growing spectrum of ALSP genetics and phenotypes enhances our knowledge of this condition, which can be crucial for both diagnosis and potential future treatments.

Quantification of tissue shrinkage and dehydration caused by microwave ablation: experimental study in kidneys for the estimation of effective coagulation volume.

PURPOSE: To quantify the extent of tissue shrinkage and dehydration caused by microwave (MW) ablation in kidneys for estimation of effective coagulation volume. MATERIALS AND METHODS: MW ablations were carried out in ex vivo porcine kidneys. Six study groups were defined: groups 1A, 2A, and 3A for MW ablation (90 W for 5 min, 7.5 min, or 10 min), and groups 1B, 2B, and 3B for control (without MW ablation). Pre- and postinterventional volume analyses were performed. Effective coagulation volumes (original tissue included in coagulation) were determined. Postinterventional dehydration analyses were performed with calculation of mean mass fractions of water. RESULTS: Mean deployed energies were 21.6 kJ ± 1.1 for group 1A, 29.9 kJ ± 1.0 for group 2A, and 42.1 kJ ± 0.5 kJ for group 3A, and were significantly different (P < .0001). Differences between pre- and postinterventional volumes were -3.8% ± 0.6 for group 1A, -5.6% ± 0.9 for group 2A, and -7.2% ± 0.4 for group 3A, and -1.1% ± 0.3 for group 1B, -1.8% ± 0.4 for group 2B, and -1.1% ± 0.4 for group 3B. Postinterventional volumes were significantly smaller than preinterventional volumes for all groups (P < .01). Underestimations of effective coagulation volume from visualized coagulation volume were 26.1% ± 3.5 for group 1A, 35.2% ± 11.2 for group 2A, and 42.1% ± 4.9 for group 3A, which were significantly different (P < .01). Mean mass fractions of water were 64.2% ± 1.4 for group 1A, 63.2% ± 1.7 for group 2A, and 62.6% ± 1.8% for group 3A, with significant differences versus corresponding control groups (P < .01). CONCLUSIONS: For MW ablation in kidneys, underestimation of effective coagulation volume based on visualized coagulation volume is significantly greater with greater deployed energy. Therefore, local dehydration with tissue shrinkage is a potential contributor.

Irreversible electroporation of the pig kidney with involvement of the renal pelvis: technical aspects, clinical outcome, and three-dimensional CT rendering for assessment of the treatment zone.

PURPOSE: To analyze irreversible electroporation (IRE) of the pig kidney with involvement of the renal pelvis. MATERIALS AND METHODS: IRE of renal tissue including the pelvis was performed in 10 kidneys in five pigs. Three study groups were defined: group I (two applicators with parallel configuration; n = 11), group II (three applicators with triangular configuration; n = 2), and group III (six applicators with complex configuration; n = 3). After IRE and before euthanasia, pigs underwent contrast-enhanced computed tomography (CT). Technical aspects (radial distance of applicators, resulting mean current), clinical outcome (complications, blood samples), and three-dimensional CT rendering for assessment of the treatment zone (short axis, circularity) were assessed. RESULTS: Radial distances of applicators were 14.3 mm ± 2.8 in group I, 12.3 mm ± 1.9 in group II, and 16.4 mm ± 3.5 in group III. Resulting mean currents were 25.7 A ± 6.5 in group I, 27.0 A ± 7.1 in group II, and 39.4 A ± 8.9 in group III. In group III, two perirenal hematomas were identified. There was no damage to the renal pelvis. During IRE, clinical blood parameters and cardiovascular markers did not change significantly. Short axis measurements were 20.6 mm ± 3.6 in group I, 31.9 mm ± 8.2 in group II, and 39.3 mm ± 2.4 in group III (P < .01 between groups). Circularity scores were 0.8 ± 0.2 in group I, 0.7 ± 0.1 in group II, and 0.7 ± 0.1 in group III, with a score of 1 indicating perfect roundness (P value not significant). CONCLUSIONS: IRE of the pig kidney with involvement of the renal pelvis is feasible and safe. Size but not shape of the treatment zone is significantly affected by applicator configuration.

Sorafenib treatment is save and may affect survival of recurrent hepatocellular carcinoma after liver transplantation.

INTRODUCTION: Recurrent hepatocellular carcinoma (HCC) after liver transplantation (LT) is a rare but challenging condition. In most cases, the recurrent tumor is presented with extrahepatic spread. Therefore, systemic treatment with sorafenib has to be assessed. Because of a plethora of possible drug interactions, e.g., with immunosuppressant or anti-infective therapy, safety and feasibility of sorafenib treatment requires special attention. MATERIALS AND METHODS: We retrospectively analyzed 18 patients who suffered from recurrent advanced HCC after LT between January 2002 and December 2010 at the University Hospital Heidelberg regarding safety of sorafenib treatment and survival. RESULTS: Results showed that 8 patients were eligible for treatment with sorafenib showing a median time to progression (TTP) of 4.5 months and an overall survival of 9 months. Most common side effects were grades I and II diarrhea and hand-foot syndrome (HFS) which could be managed by sorafenib dose reduction. No grade III or IV adverse events (AEs) were noticed. No patient had to discontinue treatment due to AEs. The ten patients not amenable for sorafenib treatment, due to initial poor performance status or its deterioration after first line treatment, were treated with surgical resection (n = 3), locoregional therapies (n = 1), or palliative radiation therapy (n = 1). They showed a median overall survival of 2.3 months. CONCLUSION: Sorafenib may represent a therapeutic option for recurrent HCC after LT with manageable side effects. The clinical benefit of sorafenib in this setting is promising but needs to be confirmed in a prospective randomized trial.

Generation of a heterozygous and a homozygous CSF1R knockout line from iPSC using CRISPR/Cas9.

Mutations in Colony-stimulating factor 1 receptor (CSF1R) lead to CSF1R-related leukoencephalopathy, also known as Adult-onset leukoencephalopathy with axonal spheroids and pigmented glia (ALSP), a rapidly progressing neurodegenerative disease with severe cognitive and motor impairment. In this study, a homozygous and a heterozygous CSF1R knockout induced pluripotent stem cell (iPSC) line were generated by CRISPR/Cas9-based gene editing. These in vitro models will provide a helpful tool for investigating the still largely unknown pathophysiology of CSF1R-related leukoencephalopathy.

Modifying UTAUT2 for a cross-country comparison of telemedicine adoption.

The ongoing COVID19 pandemic has put digital health technologies in the spotlight. To gain a deeper understanding of patients' usage intentions of virtual doctor appointments, the present research adapts the Unified Theory of Acceptance and Use of Technology 2 (UTAUT2) by integrating perceived security and perceived product advantage, two known barriers to successful telemedicine adoption. Applying age-stratified sampling, an online survey was distributed to 800 citizens in Germany and the United States of America. 710 completed and valid questionnaires were subsequently analyzed using SPSS and AMOS (versions 24). Significant, direct, and positive effects of performance expectancy, hedonic motivation, perceived security, and perceived product advantage on the behavioral intention to use virtual doctor appointments were found. The analysis of the moderating variables, age and gender, showed significant differences in user's performance expectancy and effort expectancy, and perceived product advantage, respectively. With virtual health care models on the rise, these results are important for stakeholders such as policymakers, governments, employers, but also physicians, and insurance companies as they offer clear recommendations to design telemedicine adoption strategies to ensure successful patient engagement.

Lysosomal acid lipase deficiency in pediatric patients: a scoping review.

OBJECTIVE: Lysosomal acid lipase deficiency (LAL-D) is an underdiagnosed autosomal recessive disease with onset between the first years of life and adulthood. Early diagnosis is crucial for effective therapy and long-term survival. The objective of this article is to recognize warning signs among the clinical and laboratory characteristics of LAL-D in pediatric patients through a scope review. SOURCES: Electronic searches in the Embase, PubMed, Livivo, LILACS, Web of Science, Scopus, Google Scholar, Open Gray, and ProQuest Dissertations and Theses databases. The dataset included observational studies with clinical and laboratory characteristics of infants, children and adolescents diagnosed with lysosomal acid lipase deficiency by enzyme activity testing or analysis of mutations in the lysosomal acid lipase gene (LIPA). The reference selection process was performed in two stages. The references were selected by two authors, and the data were extracted in June 2020. SUMMARY OF THE FINDINGS: The initial search returned 1593 studies, and the final selection included 108 studies from 30 countries encompassing 206 patients, including individuals with Wolman disease and cholesteryl ester storage disease (CESD). The most prevalent manifestations in both spectra of the disease were hepatomegaly, splenomegaly, anemia, dyslipidemia, and elevated transaminases. CONCLUSIONS: Vomiting, diarrhea, jaundice, and splenomegaly may be correlated, and may serve as a starting point for investigating LAL-D. Familial lymphohistiocytosis should be part of the differential diagnosis with LAL-D, and all patients undergoing upper gastrointestinal endoscopy should be submitted to intestinal biopsy.

Developmental Neurotoxicity of Fipronil and Rotenone on a Human Neuronal In Vitro Test System.

Pesticide exposure during in utero and early postnatal development can cause a wide range of neurological defects. However, relatively few insecticides have been recognized as developmental neurotoxicants, so far. Recently, discovery of the insecticide, fipronil, in chicken eggs has raised public concern. The status of fipronil as a potential developmental neurotoxicant is still under debate. Whereas several in vivo and in vitro studies suggest specific toxicity, other in vitro studies could not confirm this concern. Here, we tested fipronil and its main metabolic product, fipronil sulfone both at concentrations between 1.98 and 62.5 µM, alongside with the established developmental neurotoxicant, rotenone (0.004-10 µM) in vitro on the human neuronal precursor cell line NT2. We found that rotenone impaired all three tested DNT endpoints, neurite outgrowth, neuronal differentiation, and precursor cell migration in a dose-dependent manner and clearly separable from general cytotoxicity in the nanomolar range. Fipronil and fipronil sulfone specifically inhibited cell migration and neuronal differentiation, but not neurite outgrowth in the micromolar range. The rho-kinase inhibitor Y-27632 counteracted inhibition of migration for all three compounds (EC50 between 12 and 50 µM). The antioxidant, n-acetyl cysteine, could ameliorate the inhibitory effects of fipronil on all three tested endpoints (EC 50 between 84 and 164 µM), indicating the involvement of oxidative stress. Fipronil sulfone had a stronger effect than fipronil, confirming the importance to test metabolic products alongside original pesticides. We conclude that in vitro fipronil and fipronil sulfone display specific developmental neurotoxicity on developing human model neurons.

NO Synthesis in Immune-Challenged Locust Hemocytes and Potential Signaling to the CNS.

Similar to vertebrates, insects are exposed to a broad variety of pathogens. The innate insect immune system provides several response mechanisms such as phagocytosis, melanization, and the synthesis of antimicrobial or cytotoxic compounds. The cytotoxic nitric oxide (NO), which is also a neurotransmitter, is involved in the response to bacterial infections in various insects but has rarely been shown to be actually produced in hemocytes. We quantified the NO production in hemocytes of Locusta migratoria challenged with diverse immune stimuli by immunolabeling the by-product of NO synthesis, citrulline. Whereas in untreated adult locusts less than 5% of circulating hemocytes were citrulline-positive, the proportion rose to over 40% after 24 hours post injection of heat-inactivated bacteria. Hemocytes surrounded and melanized bacteria in locust nymphs by forming capsules. Such sessile hemocytes also produced NO. As in other insect species, activated hemocytes were found dorsally, close to the heart. In addition, we frequently observed citrulline-positive hemocytes and capsules near the ventral nerve cord. Neurites in the CNS of sterile locust embryos responded with elevation of the second messenger cGMP after contact with purified adult NO-producing hemocytes as revealed by immunofluorescence. We suggest that hemocytes can mediate a response in the CNS of an infected animal via the NO/cGMP signaling pathway.