Slow rewarming after hypothermia does not ameliorate white matter injury after hypoxia-ischemia in near-term fetal sheep.

BACKGROUND: The optimal rate to rewarm infants after therapeutic hypothermia is unclear. In this study we examined whether slow rewarming after 72 h of hypothermia would attenuate white matter injury. METHODS: Near-term fetal sheep received sham occlusion (n = 8) or cerebral ischemia for 30 min, followed by normothermia (n = 7) or hypothermia from 3-72 h, with either spontaneous fast rewarming (n = 8) within 1 h, or slow rewarming at ~0.5 °C/h (n = 8) over 10 h. Fetuses were euthanized 7 days later. RESULTS: Ischemia was associated with loss of total and mature oligodendrocytes, reduced expression of myelin proteins and induction of microglia and astrocytes, compared with sham controls (P < 0.05). Both hypothermia protocols were associated with a significant increase in numbers of total and mature oligodendrocytes, area fraction of myelin proteins and reduced numbers of microglia and astrocytes, compared with ischemia-normothermia (P < 0.05). There was no difference in the number of oligodendrocytes, microglia or astrocytes or expression of myelin proteins between fast and slow rewarming after hypothermia. CONCLUSION: The rate of rewarming after a clinically relevant duration of hypothermia had no apparent effect on white matter protection by hypothermia after cerebral ischemia in near-term fetal sheep. IMPACT: Persistent white matter injury is a major contributor to long-term disability after neonatal encephalopathy despite treatment with therapeutic hypothermia. The optimal rate to rewarm infants after therapeutic hypothermia is unclear; current protocols were developed on a precautionary basis. We now show that slow rewarming at 0.5 °C/h did not improve histological white matter injury compared with rapid spontaneous rewarming after a clinically established duration of hypothermia in near-term fetal sheep.

Do patients with central sensitivity syndromes have poor subjective outcomes despite anatomical cure from pelvic organ prolapse surgery?

INTRODUCTION: Pelvic organ prolapse(POP) has an adverse impact on quality of life with lifetime risk of surgery varying from 11 to 20%. Conditions such as fibromyalgia (FMS), chronic fatigue syndrome (CFS/ME) and irritable bowel syndrome (IBS), collectively known as central sensitivity syndromes (CSS), may affect the outcome of POP surgery. The aim of this article is to compare the outcomes of vaginal POP surgery between women with and without CSS. METHOD: This was a prospective cohort study. The validated Central Sensitisation Inventory (CSI) was used to identify women with CSS. Subjective and objective outcomes were compared between the two groups using POP-SS, Expectation and satisfaction/"EGGS", pain scores and the POP quantification system (POP-Q). A non-parametric test was used for analysis. RESULT: Seventy-eight women were recruited. Complete data were available in 62 patients; 23 patients had evidence of CSS and 39 did not. Women with CSS had significantly higher pre- and post-operative POP-SS scores than those without (p < 0.0005, p = 0.004). Seventeen (73.9%) women with CSS compared to 38 (97.4%) women without CSS demonstrated improvement of a minimum 6 points on the POP-SS scale; however, this was not stastically significant. McGill's pain scores were higher in women with CSS both pre- and post-surgery. Ninety-five per cent of women without CSS achieved their goals and were satisfied with the surgery compared to 69.5% of women with CSS (p < 005). CONCLUSION: There is a less favourable outcome of POP surgery in women with CSS compared to those without in terms of persistence of symptoms, pain and overall satisfaction.

Correction to: Vascular endothelial growth factor is an autocrine growth factor, signaling through neuropilin-1 in non-small cell lung cancer.

Since the publication of this work [1] and in response to a recent query that was brought to our attention in relation to the Western Blot in Figure 1(C) for NP2, protein lysates prepared around the same time as those presented in the manuscript in question, were run by SDS-PAGE under similar experimental conditions and probed using the same primary antibodies to NP1 and NP2 that were used originally.

The proportion of women with central sensitivity syndrome in gynecology outpatient clinics (GOPDs).

INTRODUCTION AND HYPOTHESIS: Patients in gynecology outpatient clinics (GOPDs) may present with symptoms that do not correlate well with the observed pathology and are usually labelled as having a functional disorder or medically unexplained symptoms (MUS). Underlying central sensitivity syndrome (CSS) with central sensitization (CS) as a potential mechanism may be responsible for some of their symptoms. The aim of this study is to identify the proportion of women with central sensitivity syndrome attending GOPDs. METHODS: This was a prospective study. All women attending a GOPD included in the study were asked to complete a validated Central Sensitization Inventory (CSI). The responses were graded on a Likert scale from 0 (never) to 4 (always). The total score ranges from 0 to 100. For screening purposes, a single CSI cutoff score of 40 was used to identify the group of women who may have central sensitization syndrome. RESULTS: Three hundred twenty-six women participated in the study. Overall, 123 (37%) women achieved a score above 40. This could be interpreted as these patients having increased risk of underlying central sensitization. Of these, 43 had a previously confirmed diagnosis of migraine, 55 (44%) depression, 39 (31.7%) anxiety, 11 fibromyalgia (FM), 34 irritable bowel syndrome (IBS) and 16 chronic fatigue syndrome (CFS/ME). CONCLUSIONS: Managing patients and their expectations in gynecological outpatient departments when symptoms are inconsistent with observable pathological findings is challenging. This is further complicated when patients have a concomitant central sensitivity syndrome, which can also influence the surgical outcome. Identifying these patients is a key factor for appropriate management.

Predicting Technical Success after Fistuloplasty: An Analysis of 176 Procedures.

BACKGROUND: Significant stenoses in arteriovenous fistulae (AVFs) or arteriovenous grafts (AVGs) with limitation of flow and dialysis inadequacy should prompt consideration for fistuloplasty. We sought to identify fistulae, lesions, and patient-specific variables, which predict for outcomes after fistuloplasty. METHODS: Data were extracted retrospectively from a renal access database from 2011 to 2016 of patients undergoing fistuloplasty. Demographics, comorbidities, outcomes of intervention, and flow rates documented on preintervention and postintervention duplex were collected. Secondary analysis of factors associated with postfistuloplasty flow rates of >600 mL/min, previously shown to be predictive of not requiring future intervention, was performed. RESULTS: Of 204 attempted fistuloplasties, 176 were completed. One hundred forty (79.5%) were native AVFs and 34 (19.3%), AVGs (no data for 2). Median stenosis treated was 75%, with a majority (43.8%) in the proximal outflow vein. Flow rate on duplex after fistuloplasty was significantly better in AVFs (mean improvement 189.2 mL/min) than that in AVGs (mean improvement 51.8 mL/min; P = 0.034). Greatest flow improvement occurred for needling site stenotic lesions compared with other locations (from anastomosis to central vein) but was not significant. Brachio-brachial or brachio-axillary AVGs did significantly (P < 0.05) worse than all other fistulae types. The presence of hypertension was predicted for postfistuloplasty flow rate of >600 mL/min. CONCLUSIONS: Flow rates after fistuloplasty vary depending on the type of fistula treated and the presence of hypertension. Knowledge of this can lead to better patient selection and counseling for fistuloplasty.

Tyrosine kinase inhibitors as modulators of trastuzumab-mediated antibody-dependent cell-mediated cytotoxicity in breast cancer cell lines.

BACKGROUND: Trastuzumab is an anti-HER2 monoclonal antibody (mAb) therapy capable of antibody-dependent cell-mediated cytotoxicity (ADCC) and used in the treatment of HER2+ breast cancer. Through interactions with FcÆ´R+ immune cell subsets, trastuzumab functions as a passive immunotherapy. The EGFR/HER2-targeting tyrosine kinase inhibitor (TKI) lapatinib and the next generation TKIs afatinib and neratinib, can alter HER2 levels, potentially modulating the ADCC response to trastuzumab. Using LDH-release assays, we investigated the impact of antigen modulation, assay duration and peripheral blood mononuclear cell (PBMC) activity on trastuzumab-mediated ADCC in breast cancer models of maximal (SKBR3) and minimal (MCF-7) target antigen expression to determine if modulating the ADCC response to trastuzumab using TKIs may be a viable approach for enhancing tumor immune reactivity. METHODS: HER2 levels were determined in lapatinib, afatinib and neratinib-treated SKBR3 and MCF-7 using high content analysis (HCA). Trastuzumab-mediated ADCC was assessed following treatment with TKIs utilising a colorimetric LDH release-based protocol at 4 and 12h timepoints. PBMC activity was assessed against non-MHC-restricted K562 cells. A flow cytometry-based method (CFSE/7-AAD) was also used to measure trastuzumab-mediated ADCC in medium-treated SKBR3 and MCF-7. RESULTS: HER2 antigen levels were significantly altered by the three TKIs in both cell line models. The TKIs significantly reduced LDH levels directly in SKBR3 cells but not MCF-7. Lapatinib and neratinib augment trastuzumab-related ADCC in SKBR3 but the effect was not consistent with antigen expression levels and was dependent on volunteer PBMC activity (vs. K562). A 12h assay timepoint produced more consistent results. Trastuzumab-mediated ADCC (PBMC:target cell ratio of 10:1) was measured at 7.6±4.7% (T12) by LDH assay and 19±3.2 % (T12) using the flow cytometry-based method in the antigen-low model MCF-7. CONCLUSIONS: In the presence of effector cells with high cytotoxic capacity, TKIs have the ability to augment the passive immunotherapeutic potential of trastuzumab in SKBR3, a model of HER2+ breast cancer. ADCC levels detected by LDH release assays are extremely low in MCF-7; the flow cytometry-based CFSE/7-AAD method is more sensitive and consistent for the determination of ADCC in HER2-low models.

Vascular endothelial growth factor is an autocrine growth factor, signaling through neuropilin-1 in non-small cell lung cancer.

BACKGROUND: The VEGF pathway has become an important therapeutic target in lung cancer, where VEGF has long been established as a potent pro-angiogenic growth factor expressed by many types of tumors. While Bevacizumab (Avastin) has proven successful in increasing the objective tumor response rate and in prolonging progression and overall survival in patients with NSCLC, the survival benefit is however relatively short and the majority of patients eventually relapse. The current use of tyrosine kinase inhibitors alone and in combination with chemotherapy has been underwhelming, highlighting an urgent need for new targeted therapies. In this study, we examined the mechanisms of VEGF-mediated survival in NSCLC cells and the role of the Neuropilin receptors in this process. METHODS: NSCLC cells were screened for expression of VEGF and its receptors. The effects of recombinant VEGF and its blockade on lung tumor cell proliferation and cell cycle were examined. Phosphorylation of Akt and Erk1/2 proteins was examined by high content analysis and confocal microscopy. The effects of silencing VEGF on cell proliferation and survival signaling were also assessed. A Neuropilin-1 stable-transfected cell line was generated. Cell growth characteristics in addition to pAkt and pErk1/2 signaling were studied in response to VEGF and its blockade. Tumor growth studies were carried out in nude mice following subcutaneous injection of NP1 over-expressing cells. RESULTS: Inhibition of the VEGF pathway with anti-VEGF and anti-VEGFR-2 antibodies or siRNA to VEGF, NP1 and NP2 resulted in growth inhibition of NP1 positive tumor cell lines associated with down-regulation of PI3K and MAPK kinase signaling. Stable transfection of NP1 negative cells with NP1 induced proliferation in vitro, which was further enhanced by exogenous VEGF. In vivo, NP1 over-expressing cells significantly increased tumor growth in xenografts compared to controls. CONCLUSIONS: Our data demonstrate that VEGF is an autocrine growth factor in NSCLC signaling, at least in part, through NP1. Targeting this VEGF receptor may offer potential as a novel therapeutic approach and also support the evaluation of the role of NP1 as a biomarker predicting sensitivity or resistance to VEGF and VEGFR-targeted therapies in the clinical arena.

L-plastin regulates polarization and migration in chemokine-stimulated human T lymphocytes.

Chemokines such as SDF-1α play a crucial role in orchestrating T lymphocyte polarity and migration via polymerization and reorganization of the F-actin cytoskeleton, but the role of actin-associated proteins in this process is not well characterized. In this study, we have investigated a role for L-plastin, a leukocyte-specific F-actin-bundling protein, in SDF-1α-stimulated human T lymphocyte polarization and migration. We found that L-plastin colocalized with F-actin at the leading edge of SDF-1α-stimulated T lymphocytes and was also phosphorylated at Ser(5), a site that when phosphorylated regulates the ability of L-plastin to bundle F-actin. L-plastin phosphorylation was sensitive to pharmacological inhibitors of protein kinase C (PKC), and several PKC isoforms colocalized with L-plastin at the leading edge of SDF-1α-stimulated lymphocytes. However, PKC ζ, an established regulator of cell polarity, was the only isoform that regulated L-plastin phosphorylation. Knockdown of L-plastin expression with small interfering RNAs demonstrated that this protein regulated the localization of F-actin at the leading edge of chemokine-stimulated cells and was also required for polarization, lamellipodia formation, and chemotaxis. Knockdown of L-plastin expression also impaired the Rac1 activation cycle and Akt phosphorylation in response to SDF-1α stimulation. Furthermore, L-plastin also regulated SDF-1α-mediated lymphocyte migration on the integrin ligand ICAM-1 by influencing velocity and persistence, but in a manner that was independent of LFA-1 integrin activation or adhesion. This study, therefore, demonstrates an important role for L-plastin and the signaling pathways that regulate its phosphorylation in response to chemokines and adds L-plastin to a growing list of proteins implicated in T lymphocyte polarity and migration.

On the nomenclatural status of type genera in Coleoptera (Insecta).

More than 4700 nominal family-group names (including names for fossils and ichnotaxa) are nomenclaturally available in the order Coleoptera. Since each family-group name is based on the concept of its type genus, we argue that the stability of names used for the classification of beetles depends on accurate nomenclatural data for each type genus. Following a review of taxonomic literature, with a focus on works that potentially contain type species designations, we provide a synthesis of nomenclatural data associated with the type genus of each nomenclaturally available family-group name in Coleoptera. For each type genus the author(s), year of publication, and page number are given as well as its current status (i.e., whether treated as valid or not) and current classification. Information about the type species of each type genus and the type species fixation (i.e., fixed originally or subsequently, and if subsequently, by whom) is also given. The original spelling of the family-group name that is based on each type genus is included, with its author(s), year, and stem. We append a list of nomenclaturally available family-group names presented in a classification scheme. Because of the importance of the Principle of Priority in zoological nomenclature, we provide information on the date of publication of the references cited in this work, when known. Several nomenclatural issues emerged during the course of this work. We therefore appeal to the community of coleopterists to submit applications to the International Commission on Zoological Nomenclature (henceforth "Commission") in order to permanently resolve some of the problems outlined here. The following changes of authorship for type genera are implemented here (these changes do not affect the concept of each type genus): CHRYSOMELIDAE: Fulcidax Crotch, 1870 (previously credited to "Clavareau, 1913"); CICINDELIDAE: Euprosopus W.S. MacLeay, 1825 (previously credited to "Dejean, 1825"); COCCINELLIDAE: Alesia Reiche, 1848 (previously credited to "Mulsant, 1850"); CURCULIONIDAE: Arachnopus Boisduval, 1835 (previously credited to "Guérin-Méneville, 1838"); ELATERIDAE: Thylacosternus Gemminger, 1869 (previously credited to "Bonvouloir, 1871"); EUCNEMIDAE: Arrhipis Gemminger, 1869 (previously credited to "Bonvouloir, 1871"), Mesogenus Gemminger, 1869 (previously credited to "Bonvouloir, 1871"); LUCANIDAE: Sinodendron Hellwig, 1791 (previously credited to "Hellwig, 1792"); PASSALIDAE: Neleides Harold, 1868 (previously credited to "Kaup, 1869"), Neleus Harold, 1868 (previously credited to "Kaup, 1869"), Pertinax Harold, 1868 (previously credited to "Kaup, 1869"), Petrejus Harold, 1868 (previously credited to "Kaup, 1869"), Undulifer Harold, 1868 (previously credited to "Kaup, 1869"), Vatinius Harold, 1868 (previously credited to "Kaup, 1869"); PTINIDAE: Mezium Leach, 1819 (previously credited to "Curtis, 1828"); PYROCHROIDAE: Agnathus Germar, 1818 (previously credited to "Germar, 1825"); SCARABAEIDAE: Eucranium Dejean, 1833 (previously "Brullé, 1838"). The following changes of type species were implemented following the discovery of older type species fixations (these changes do not pose a threat to nomenclatural stability): BOLBOCERATIDAE: Bolbocerusbocchus Erichson, 1841 for Bolbelasmus Boucomont, 1911 (previously Bolbocerasgallicum Mulsant, 1842); BUPRESTIDAE: Stigmoderaguerinii Hope, 1843 for Neocuris Saunders, 1868 (previously Anthaxiafortnumi Hope, 1846), Stigmoderaperoni Laporte & Gory, 1837 for Curis Laporte & Gory, 1837 (previously Buprestiscaloptera Boisduval, 1835); CARABIDAE: Carabuselatus Fabricius, 1801 for Molops Bonelli, 1810 (previously Carabusterricola Herbst, 1784 sensu Fabricius, 1792); CERAMBYCIDAE: Prionuspalmatus Fabricius, 1792 for Macrotoma Audinet-Serville, 1832 (previously Prionusserripes Fabricius, 1781); CHRYSOMELIDAE: Donaciaequiseti Fabricius, 1798 for Haemonia Dejean, 1821 (previously Donaciazosterae Fabricius, 1801), Eumolpusruber Latreille, 1807 for Euryope Dalman, 1824 (previously Cryptocephalusrubrifrons Fabricius, 1787), Galerucaaffinis Paykull, 1799 for Psylliodes Latreille, 1829 (previously Chrysomelachrysocephala Linnaeus, 1758); COCCINELLIDAE: Dermestesrufus Herbst, 1783 for Coccidula Kugelann, 1798 (previously Chrysomelascutellata Herbst, 1783); CRYPTOPHAGIDAE: Ipscaricis G.-A. Olivier, 1790 for Telmatophilus Heer, 1841 (previously Cryptophagustyphae Fallén, 1802), Silphaevanescens Marsham, 1802 for Atomaria Stephens, 1829 (previously Dermestesnigripennis Paykull, 1798); CURCULIONIDAE: Bostrichuscinereus Herbst, 1794 for Crypturgus Erichson, 1836 (previously Bostrichuspusillus Gyllenhal, 1813); DERMESTIDAE: Dermestestrifasciatus Fabricius, 1787 for Attagenus Latreille, 1802 (previously Dermestespellio Linnaeus, 1758); ELATERIDAE: Elatersulcatus Fabricius, 1777 for Chalcolepidius Eschscholtz, 1829 (previously Chalcolepidiuszonatus Eschscholtz, 1829); ENDOMYCHIDAE: Endomychusrufitarsis Chevrolat, 1835 for Epipocus Chevrolat, 1836 (previously Endomychustibialis Guérin-Méneville, 1834); EROTYLIDAE: Ipshumeralis Fabricius, 1787 for Dacne Latreille, 1797 (previously Dermestesbipustulatus Thunberg, 1781); EUCNEMIDAE: Fornaxaustrocaledonicus Perroud & Montrouzier, 1865 for Mesogenus Gemminger, 1869 (previously Mesogenusmellyi Bonvouloir, 1871); GLAPHYRIDAE: Melolonthaserratulae Fabricius, 1792 for Glaphyrus Latreille, 1802 (previously Scarabaeusmaurus Linnaeus, 1758); HISTERIDAE: Histerstriatus Forster, 1771 for Onthophilus Leach, 1817 (previously Histersulcatus Moll, 1784); LAMPYRIDAE: Ototretafornicata E. Olivier, 1900 for Ototreta E. Olivier, 1900 (previously Ototretaweyersi E. Olivier, 1900); LUCANIDAE: Lucanuscancroides Fabricius, 1787 for Lissotes Westwood, 1855 (previously Lissotesmenalcas Westwood, 1855); MELANDRYIDAE: Nothusclavipes G.-A. Olivier, 1812 for Nothus G.-A. Olivier, 1812 (previously Nothuspraeustus G.-A. Olivier, 1812); MELYRIDAE: Lagriaater Fabricius, 1787 for Enicopus Stephens, 1830 (previously Dermesteshirtus Linnaeus, 1767); NITIDULIDAE: Sphaeridiumluteum Fabricius, 1787 for Cychramus Kugelann, 1794 (previously Strongylusquadripunctatus Herbst, 1792); OEDEMERIDAE: Helopslaevis Fabricius, 1787 for Ditylus Fischer, 1817 (previously Ditylushelopioides Fischer, 1817 [sic]); PHALACRIDAE: Sphaeridiumaeneum Fabricius, 1792 for Olibrus Erichson, 1845 (previously Sphaeridiumbicolor Fabricius, 1792); RHIPICERIDAE: Sandalusniger Knoch, 1801 for Sandalus Knoch, 1801 (previously Sandaluspetrophya Knoch, 1801); SCARABAEIDAE: Cetoniaclathrata G.-A. Olivier, 1792 for Inca Lepeletier & Audinet-Serville, 1828 (previously Cetoniaynca Weber, 1801); Gnathoceravitticollis W. Kirby, 1825 for Gnathocera W. Kirby, 1825 (previously Gnathoceraimmaculata W. Kirby, 1825); Melolonthavillosula Illiger, 1803 for Chasmatopterus Dejean, 1821 (previously Melolonthahirtula Illiger, 1803); STAPHYLINIDAE: Staphylinuspolitus Linnaeus, 1758 for Philonthus Stephens, 1829 (previously Staphylinussplendens Fabricius, 1792); ZOPHERIDAE: Hispamutica Linnaeus, 1767 for Orthocerus Latreille, 1797 (previously Tenebriohirticornis DeGeer, 1775). The discovery of type species fixations that are older than those currently accepted pose a threat to nomenclatural stability (an application to the Commission is necessary to address each problem): CANTHARIDAE: Malthinus Latreille, 1805, Malthodes Kiesenwetter, 1852; CARABIDAE: Bradycellus Erichson, 1837, Chlaenius Bonelli, 1810, Harpalus Latreille, 1802, Lebia Latreille, 1802, Pheropsophus Solier, 1834, Trechus Clairville, 1806; CERAMBYCIDAE: Callichroma Latreille, 1816, Callidium Fabricius, 1775, Cerasphorus Audinet-Serville, 1834, Dorcadion Dalman, 1817, Leptura Linnaeus, 1758, Mesosa Latreille, 1829, Plectromerus Haldeman, 1847; CHRYSOMELIDAE: Amblycerus Thunberg, 1815, Chaetocnema Stephens, 1831, Chlamys Knoch, 1801, Monomacra Chevrolat, 1836, Phratora Chevrolat, 1836, Stylosomus Suffrian, 1847; COLONIDAE: Colon Herbst, 1797; CURCULIONIDAE: Cryphalus Erichson, 1836, Lepyrus Germar, 1817; ELATERIDAE: Adelocera Latreille, 1829, Beliophorus Eschscholtz, 1829; ENDOMYCHIDAE: Amphisternus Germar, 1843, Dapsa Latreille, 1829; GLAPHYRIDAE: Anthypna Eschscholtz, 1818; HISTERIDAE: Hololepta Paykull, 1811, Trypanaeus Eschscholtz, 1829; LEIODIDAE: Anisotoma Panzer, 1796, Camiarus Sharp, 1878, Choleva Latreille, 1797; LYCIDAE: Calopteron Laporte, 1838, Dictyoptera Latreille, 1829; MELOIDAE: Epicauta Dejean, 1834; NITIDULIDAE: Strongylus Herbst, 1792; SCARABAEIDAE: Anisoplia Schönherr, 1817, Anticheira Eschscholtz, 1818, Cyclocephala Dejean, 1821, Glycyphana Burmeister, 1842, Omaloplia Schönherr, 1817, Oniticellus Dejean, 1821, Parachilia Burmeister, 1842, Xylotrupes Hope, 1837; STAPHYLINIDAE: Batrisus Aubé, 1833, Phloeonomus Heer, 1840, Silpha Linnaeus, 1758; TENEBRIONIDAE: Bolitophagus Illiger, 1798, Mycetochara Guérin-Méneville, 1827. Type species are fixed for the following nominal genera: ANTHRIBIDAE: Decataphanesgracilis Labram & Imhoff, 1840 for Decataphanes Labram & Imhoff, 1840; CARABIDAE: Feroniaerratica Dejean, 1828 for Loxandrus J.L. LeConte, 1853; CERAMBYCIDAE: Tmesisternusoblongus Boisduval, 1835 for Icthyosoma Boisduval, 1835; CHRYSOMELIDAE: Brachydactylaannulipes Pic, 1913 for Pseudocrioceris Pic, 1916, Cassidaviridis Linnaeus, 1758 for Evaspistes Gistel, 1856, Ocnosceliscyanoptera Erichson, 1847 for Ocnoscelis Erichson, 1847, Promecothecapetelii Guérin-Méneville, 1840 for Promecotheca Guérin- Méneville, 1840; CLERIDAE: Attelabusmollis Linnaeus, 1758 for Dendroplanetes Gistel, 1856; CORYLOPHIDAE: Corylophusmarginicollis J.L. LeConte, 1852 for Corylophodes A. Matthews, 1885; CURCULIONIDAE: Hoplorhinusmelanocephalus Chevrolat, 1878 for Hoplorhinus Chevrolat, 1878; SonnetiusbinariusCasey, 1922 for Sonnetius Casey, 1922; ELATERIDAE: Pyrophorusmelanoxanthus Candèze, 1865 for Alampes Champion, 1896; PHYCOSECIDAE: Phycosecislitoralis Pascoe, 1875 for Phycosecis Pascoe, 1875; PTILODACTYLIDAE: Aploglossasallei Guérin-Méneville, 1849 for Aploglossa Guérin-Méneville, 1849, Coloboderaovata Klug, 1837 for Colobodera Klug, 1837; PTINIDAE: Dryophilusanobioides Chevrolat, 1832 for Dryobia Gistel, 1856; SCARABAEIDAE: Achloahelvola Erichson, 1840 for Achloa Erichson, 1840, Camentaobesa Burmeister, 1855 for Camenta Erichson, 1847, Pinotustalaus Erichson, 1847 for Pinotus Erichson, 1847, Psilonychusecklonii Burmeister, 1855 for Psilonychus Burmeister, 1855. New replacement name: CERAMBYCIDAE: Basorus Bouchard & Bousquet, nom. nov. for Sobarus Harold, 1879. New status: CARABIDAE: KRYZHANOVSKIANINI Deuve, 2020, stat. nov. is given the rank of tribe instead of subfamily since our classification uses the rank of subfamily for PAUSSINAE rather than family rank; CERAMBYCIDAE: Amymoma Pascoe, 1866, stat. nov. is used as valid over Neoamymoma Marinoni, 1977, Holopterus Blanchard, 1851, stat. nov. is used as valid over Proholopterus Monné, 2012; CURCULIONIDAE: Phytophilus Schönherr, 1835, stat. nov. is used as valid over the unnecessary new replacement name Synophthalmus Lacordaire, 1863; EUCNEMIDAE: Nematodinus Lea, 1919, stat. nov. is used as valid instead of Arrhipis Gemminger, 1869, which is a junior homonym. Details regarding additional nomenclatural issues that still need to be resolved are included in the entry for each of these type genera: BOSTRICHIDAE: Lyctus Fabricius, 1792; BRENTIDAE: Trachelizus Dejean, 1834; BUPRESTIDAE: Pristiptera Dejean, 1833; CANTHARIDAE: Chauliognathus Hentz, 1830, Telephorus Schäffer, 1766; CARABIDAE: Calathus Bonelli, 1810, Cosnania Dejean, 1821, Dicrochile Guérin-Méneville, 1847, Epactius D.H. Schneider, 1791, Merismoderus Westwood, 1847, Polyhirma Chaudoir, 1850, Solenogenys Westwood, 1860, Zabrus Clairville, 1806; CERAMBYCIDAE: Ancita J. Thomson, 1864, Compsocerus Audinet-Serville, 1834, Dorcadodium Gistel, 1856, Glenea Newman, 1842; Hesperophanes Dejean, 1835, Neoclytus J. Thomson, 1860, Phymasterna Laporte, 1840, Tetrops Stephens, 1829, Zygocera Erichson, 1842; CHRYSOMELIDAE: Acanthoscelides Schilsky, 1905, Corynodes Hope, 1841, Edusella Chapuis, 1874; Hemisphaerota Chevrolat, 1836; Physonota Boheman, 1854, Porphyraspis Hope, 1841; CLERIDAE: Dermestoides Schäffer, 1777; COCCINELLIDAE: Hippodamia Chevrolat, 1836, Myzia Mulsant, 1846, Platynaspis L. Redtenbacher, 1843; CURCULIONIDAE: Coeliodes Schönherr, 1837, Cryptoderma Ritsema, 1885, Deporaus Leach, 1819, Epistrophus Kirsch, 1869, Geonemus Schönherr, 1833, Hylastes Erichson, 1836; DYTISCIDAE: Deronectes Sharp, 1882, Platynectes Régimbart, 1879; EUCNEMIDAE: Dirhagus Latreille, 1834; HYBOSORIDAE: Ceratocanthus A. White, 1842; HYDROPHILIDAE: Cyclonotum Erichson, 1837; LAMPYRIDAE: Luciola Laporte, 1833; LEIODIDAE: Ptomaphagus Hellwig, 1795; LUCANIDAE: Leptinopterus Hope, 1838; LYCIDAE: Cladophorus Guérin-Méneville, 1830, Mimolibnetis Kazantsev, 2000; MELOIDAE: Mylabris Fabricius, 1775; NITIDULIDAE: Meligethes Stephens, 1829; PTILODACTYLIDAE: Daemon Laporte, 1838; SCARABAEIDAE: Allidiostoma Arrow, 1940, Heterochelus Burmeister, 1844, Liatongus Reitter, 1892, Lomaptera Gory & Percheron, 1833, Megaceras Hope, 1837, Stenotarsia Burmeister, 1842; STAPHYLINIDAE: Actocharis Fauvel, 1871, Aleochara Gravenhorst, 1802; STENOTRACHELIDAE: Stenotrachelus Berthold, 1827; TENEBRIONIDAE: Cryptochile Latreille, 1828, Heliopates Dejean, 1834, Helops Fabricius, 1775. First Reviser actions deciding the correct original spelling: CARABIDAE: Aristochroodes Marcilhac, 1993 (not Aritochroodes); CERAMBYCIDAE: Dorcadodium Gistel, 1856 (not Dorcadodion), EVODININI Zamoroka, 2022 (not EVODINIINI); CHRYSOMELIDAE: Caryopemon Jekel, 1855 (not Carpopemon), Decarthrocera Laboissière, 1937 (not Decarthrocerina); CICINDELIDAE: Odontocheila Laporte, 1834 (not Odontacheila); CLERIDAE: CORMODINA Bartlett, 2021 (not CORMODIINA), Orthopleura Spinola, 1845 (not Orthoplevra, not Orthopleuva); CURCULIONIDAE: Arachnobas Boisduval, 1835 (not Arachnopus), Palaeocryptorhynchus Poinar, 2009 (not Palaeocryptorhynus); DYTISCIDAE: Ambarticus Yang et al., 2019 and AMBARTICINI Yang et al., 2019 (not Ambraticus, not AMBRATICINI); LAMPYRIDAE: Megalophthalmus G.R. Gray, 1831 (not Megolophthalmus, not Megalopthalmus); SCARABAEIDAE: Mentophilus Laporte, 1840 (not Mintophilus, not Minthophilus), Pseudadoretusdilutellus Semenov, 1889 (not P.ditutellus). While the correct identification of the type species is assumed, in some cases evidence suggests that species were misidentified when they were fixed as the type of a particular nominal genus. Following the requirements of Article 70.3.2 of the International Code of Zoological Nomenclature we hereby fix the following type species (which in each case is the taxonomic species actually involved in the misidentification): ATTELABIDAE: Rhynchitescavifrons Gyllenhal, 1833 for Lasiorhynchites Jekel, 1860; BOSTRICHIDAE: Ligniperdaterebrans Pallas, 1772 for Apate Fabricius, 1775; BRENTIDAE: Ceocephalusappendiculatus Boheman, 1833 for Uroptera Berthold, 1827; BUPRESTIDAE: Buprestisundecimmaculata Herbst, 1784 for Ptosima Dejean, 1833; CARABIDAE: Amaralunicollis Schiødte, 1837 for Amara Bonelli, 1810, Buprestisconnexus Geoffroy, 1785 for Polistichus Bonelli, 1810, Carabusatrorufus Strøm, 1768 for Patrobus Dejean, 1821, Carabusgigas Creutzer, 1799 for Procerus Dejean, 1821, Carabusteutonus Schrank, 1781 for Stenolophus Dejean, 1821, Carenumbonellii Westwood, 1842 for Carenum Bonelli, 1813, Scaritespicipes G.-A. Olivier, 1795 for Acinopus Dejean, 1821, Trigonotomaindica Brullé, 1834 for Trigonotoma Dejean, 1828; CERAMBYCIDAE: Cerambyxlusitanus Linnaeus, 1767 for Exocentrus Dejean, 1835, Clytussupernotatus Say, 1824 for Psenocerus J.L. LeConte, 1852; CICINDELIDAE: Ctenostomajekelii Chevrolat, 1858 for Ctenostoma Klug, 1821; CURCULIONIDAE: Cnemogonuslecontei Dietz, 1896 for Cnemogonus J.L. LeConte, 1876; Phloeophagusturbatus Schönherr, 1845 for Phloeophagus Schönherr, 1838; GEOTRUPIDAE: Lucanusapterus Laxmann, 1770 for Lethrus Scopoli, 1777; HISTERIDAE: Histerrugiceps Duftschmid, 1805 for Hypocaccus C.G. Thomson, 1867; HYBOSORIDAE: Hybosorusilligeri Reiche, 1853 for Hybosorus W.S. MacLeay, 1819; HYDROPHILIDAE: Hydrophilusmelanocephalus G.-A. Olivier, 1793 for Enochrus C.G. Thomson, 1859; MYCETAEIDAE: Dermestessubterraneus Fabricius, 1801 for Mycetaea Stephens, 1829; SCARABAEIDAE: Aulaciumcarinatum Reiche, 1841 for Mentophilus Laporte, 1840, Phanaeusvindex W.S. MacLeay, 1819 for Phanaeus W.S. MacLeay, 1819, Ptinusgermanus Linnaeus, 1767 for Rhyssemus Mulsant, 1842, Scarabaeuslatipes Guérin-Méneville, 1838 for Cheiroplatys Hope, 1837; STAPHYLINIDAE: Scydmaenustarsatus P.W.J. Müller & Kunze, 1822 for Scydmaenus Latreille, 1802. New synonyms: CERAMBYCIDAE: CARILIINI Zamoroka, 2022, syn. nov. of ACMAEOPINI Della Beffa, 1915, DOLOCERINI Özdikmen, 2016, syn. nov. of BRACHYPTEROMINI Sama, 2008, PELOSSINI Tavakilian, 2013, syn. nov. of LYGRINI Sama, 2008, PROHOLOPTERINI Monné, 2012, syn. nov. of HOLOPTERINI Lacordaire, 1868.

Leukocyte function-associated antigen-1/intercellular adhesion molecule-1 interaction induces a novel genetic signature resulting in T-cells refractory to transforming growth factor-ß signaling.

The immunesuppressive cytokine TGF-ß plays crucial regulatory roles in the induction and maintenance of immunologic tolerance and prevention of immunopathologies. However, it remains unclear how circulating T-cells can escape from the quiescent state maintained by TGF-ß. Here, we report that the T-cell integrin leukocyte function-associated antigen-1 (LFA-1) interaction with its ligand intercellular adhesion molecule-1 (ICAM-1) induces a genetic signature associated with reduced TGF-ß responsiveness via up-regulation of SKI, E3 ubiquitin-protein ligase SMURF2, and SMAD7 (mothers against decapentaplegic homolog 7) genes and proteins. We confirmed that the expression of these TGF-ß inhibitory molecules was dependent on STAT3 and/or JNK activation. Increased expression of SMAD7 and SMURF2 in LFA-1/ICAM-1 cross-linked T-cells resulted in impaired TGF-ß-mediated phosphorylation of SMAD2 and suppression of IL-2 secretion. Expression of SKI caused resistance to TGF-ß-mediated suppression of IL-2, but SMAD2 phosphorylation was unaffected. Blocking LFA-1 by neutralizing antibody or specific knockdown of TGF-ß inhibitory molecules by siRNA substantially restored LFA-1/ICAM-1-mediated alteration in TGF-ß signaling. LFA-1/ICAM-1-stimulated human and mouse T-cells were refractory to TGF-ß-mediated induction of FOXP3(+) (forkhead box P3) and RORγt(+) (retinoic acid-related orphan nuclear receptor γt) Th17 differentiation. These mechanistic data suggest an important role for LFA-1/ICAM-1 interactions in immunoregulation concurrent with lymphocyte migration that may have implications at the level of local inflammatory response and for anti-LFA-1-based therapies.

The alpha2delta subunits of voltage-gated calcium channels form GPI-anchored proteins, a posttranslational modification essential for function.

Voltage-gated calcium channels are thought to exist in the plasma membrane as heteromeric proteins, in which the alpha1 subunit is associated with two auxiliary subunits, the intracellular beta subunit and the alpha(2)delta subunit; both of these subunits influence the trafficking and properties of Ca(V)1 and Ca(V)2 channels. The alpha(2)delta subunits have been described as type I transmembrane proteins, because they have an N-terminal signal peptide and a C-terminal hydrophobic and potentially transmembrane region. However, because they have very short C-terminal cytoplasmic domains, we hypothesized that the alpha(2)delta proteins might be associated with the plasma membrane through a glycosylphosphatidylinositol (GPI) anchor attached to delta rather than a transmembrane domain. Here, we provide biochemical, immunocytochemical, and mutational evidence to show that all of the alpha(2)delta subunits studied, alpha(2)delta-1, alpha(2)delta-2, and alpha(2)delta-3, show all of the properties expected of GPI-anchored proteins, both when heterologously expressed and in native tissues. They are substrates for prokaryotic phosphatidylinositol-phospholipase C (PI-PLC) and trypanosomal GPI-PLC, which release the alpha(2)delta proteins from membranes and intact cells and expose a cross-reacting determinant epitope. PI-PLC does not affect control transmembrane or membrane-associated proteins. Furthermore, mutation of the predicted GPI-anchor sites markedly reduced plasma membrane and detergent-resistant membrane localization of alpha(2)delta subunits. We also show that GPI anchoring of alpha(2)delta subunits is necessary for their function to enhance calcium currents, and PI-PLC treatment only reduces calcium current density when alpha(2)delta subunits are coexpressed. In conclusion, this study redefines our understanding of alpha(2)delta subunits, both in terms of their role in calcium-channel function and other roles in synaptogenesis.

Pre-Treatment of platinum resistant ovarian cancer cells with an MMP-9/MMP-2 inhibitor prior to cisplatin enhances cytotoxicity as determined by high content screening.

Platinum resistance is a major cause of treatment failure in ovarian cancer. We previously identified matrix metalloproteinase 9 (MMP-9) as a potential therapeutic target of chemoresistant disease. A2780cis (cisplatin-resistant) and A2780 (cisplatin-sensitive) ovarian carcinoma cell lines were used. The cytotoxic effect of MMP-9/MMP-2 inhibitor, (2R)-2-[(4-Biphenylsulfonyl) amino]-3 phenylpropionic acid (C21H19NO4S) alone or in combination with cisplatin was determined using high content screening. Protein expression was examined using immunohistochemistry and ELISA. Co-incubation of cisplatin and an MMP-9/MMP-2 inhibitor, (2R)-2-[(4-Biphenylsulfonyl) amino]-3 phenylpropionic acid (C21H19NO4S) resulted in significantly greater cytotoxicity as compared to either treatment alone in a cisplatin resistant MMP-9 overexpressing cell line; A2780cis. In addition, pre-incubating with MMP-9i prior to cisplatin further enhances the cytotoxic effect. No significant difference was observed in MMP-9 protein in tissue but a trend towards increased MMP-9 was observed in recurrent serum. We propose that MMP-9/MMP-2i may be utilized in the treatment of recurrent/chemoresistant ovarian cancers that overexpress MMP-9 mRNA but its role in vivo remains to be evaluated.

Aleocharine rove beetles of New Hampshire, USA: new taxa and new records (Coleoptera, Staphylinidae, Aleocharinae).

Based on the specimens housed primarily in the University of New Hampshire Insect Collection (UNH) and the Canadian National Collection (CNC), we present here a comprehensive faunal review of aleocharine beetles of the state and provide new distribution and natural history data. We report 252 species from New Hampshire belonging to some 74 genera in 15 tribes; 159 of these constitute new New Hampshire state records (NSR), of which 37 (excluding new species and including 1 New York record) constitute new country distribution records (NCR) for the USA. In addition, we provide 36 new state records for ME, with 5 of these species not yet known from NH, seven new state records for MA, two new state records for PA and VT, and one each for CT, DE, MI, NC, NY (also a NCR), OH, and OK. One new genus, Belladonna Klimaszewski and Chandler is erected, and nine species are described as new to science (alphabetical order): Agaricomorpha hampshirensis Klimaszewski and Chandler, sp. n., Atheta ellisi Klimaszewski and Chandler sp. n, Atheta monroe Klimaszewski and Chandler, sp. n., Atheta struyvei Klimaszewski and Chandler sp. n, Belladonna barryi Klimaszewski and Chandler, sp. n., Belladonna fortieri Klimaszewski and Chandler, sp. n., Colusa smetanai Klimaszewski and Chandler, sp. n., Philhygra pinkhami Klimaszewski and Chandler sp. n., and P. pseudomagniceps Klimaszewski and Chandler, sp. n. Undescribed females of Aleochara daviesi Klimaszewski and Brunke, and Silusa langori Klimaszewski, are described and illustrated. Illustrations of Atheta (Tetropla) tubericauda Bernhauer are provided for the first time, based on a male from New Hampshire. A new combination is proposed for Atheta (Dimetrota) mcalpinei Klimaszewski and Webster.

Body reprogramming for fibromyalgia and central sensitivity syndrome: A preliminary evaluation.

Objectives: Central sensitivity syndrome disorders such as fibromyalgia, provoke continued debate, highlighting diagnostic and therapeutic uncertainty. The Hyland model provides a way of understanding and treating the medically unexplained symptoms of central sensitivity syndromes using complexity theory and principles of adaption in network systems. The body reprogramming is a multi-modal intervention based on the Hyland model designed for patients living with medically unexplained symptoms. This preliminary, naturalistic and single-arm service evaluation set out to evaluate outcome after attending a body reprogramming course in patients living with fibromyalgia or central sensitivity syndrome. Methods: Patients diagnosed with fibromyalgia or central sensitivity syndrome were recruited. The body reprogramming courses consisting of eight sessions, each 2.5 h in length, were run at two study sites in England. Data were collected at baseline, post course and 3-months post course using questionnaires assessing symptomatology (FIQR/SIQR), Depression (PHQ9), Anxiety (GAD7) and quality of life (GQoL). Repeated measures t-tests were used, and all comparisons were conducted on an intention to treat basis. Results: In total, 198 patients with a mean age of 47.73 years were enrolled on the body reprogramming courses. Statistically and clinically significant improvement were observed in the FIQR from baseline to post course (mean change: 11.28) and baseline to follow-up (mean change: 15.09). PHQ9 scores also improved significantly from baseline to post course (mean reduction 3.72) and baseline to follow-up (mean reduction 5.59). Conclusions: Our study provides first evidence that the body reprogramming intervention is an effective approach for patients living with fibromyalgia or central sensitivity syndromes on a variety of clinical measures. Besides these promising results, important limitations of the study are discussed, and larger randomized controlled trials are clearly warranted.

A tumour-spheroid manufacturing and cryopreservation process that yields a highly reproducible product ready for direct use in drug screening assays.

If it were possible to purchase tumour-spheroids as a standardised product, ready for direct use in assays, this may contribute to greater research reproducibility, potentially reducing costs and accelerating outcomes. Herein, we describe a workflow where uniformly sized cancer tumour-spheroids are mass-produced using microwell culture, cryopreserved with high viability, and then cultured in neutral buoyancy media for drug testing. C4-2B prostate cancer or MCF-7 breast cancer cells amalgamated into uniform tumour-spheroids after 48 h of culture. Tumour-spheroids formed from 100 cells each tolerated the cryopreservation process marginally better than tumour-spheroids formed from 200 or 400 cells. Post-thaw, tumour-spheroid metabolic activity was significantly reduced, suggesting mitochondrial damage. Metabolic function was rescued by thawing the tumour-spheroids into medium supplemented with 10 µM N-Acetyl-l-cysteine (NAC). Following thaw, the neutral buoyancy media, Happy Cell ASM, was used to maintain tumour-spheroids as discrete tissues during drug testing. Fresh and cryopreserved C4-2B or MCF-7 tumour-spheroids responded similarly to titrations of Docetaxel. This protocol will contribute to a future where tumour-spheroids may be available for purchase as reliable and reproducible products, allowing laboratories to efficiently replicate and build on published research, in many cases, making tumour-spheroids simply another cell culture reagent.

Synthesis and biological evaluation of a library of glycoporphyrin compounds.

A library of glycosylated porphyrins (glycoporphyrins) was prepared and the compounds were evaluated for their photodynamic therapy (PDT) activity against the oesophageal squamous-cell carcinoma cell line OE21 in vitro. A synthetic methodology was developed to allow incorporation of biologically active carbohydrates, including the histo-blood-group antigen trisaccharide Lewis(X), onto the porphyrin backbone. The effect of the carbohydrate group and substitution pattern on the PDT activity, cell uptake and subcellular localisation of the glycoporphyrin compounds is reported.

Cytoskeletal re-arrangement in TGF-ß1-induced alveolar epithelial-mesenchymal transition studied by atomic force microscopy and high-content analysis.

Epithelial-mesenchymal transition (EMT) is closely implicated in the pathogenesis of idiopathic pulmonary fibrosis. Associated with this phenotypic transition is the acquisition of an elongated cell morphology and establishment of stress fibers. The extent to which these EMT-associated changes influence cellular mechanics is unclear. We assessed the biomechanical properties of alveolar epithelial cells (A549) following exposure to TGF-ß1. Using atomic force microscopy, changes in cell stiffness and surface membrane features were determined. Stimulation with TGF-ß1 gave rise to a significant increase in stiffness, which was augmented by a collagen I matrix. Additionally, TGF-ß1-treated cells exhibited a rougher surface profile with notable protrusions. Simultaneous quantitative examination of the morphological attributes of stimulated cells using an image-based high-content analysis system revealed dramatic alterations in cell shape, F-actin content and distribution. Together, these investigations point to a strong correlation between the cytoskeletal-associated cellular architecture and the mechanical dynamics of alveolar epithelial cells undergoing EMT. From the Clinical Editor: Epithelial-mesenchymal transition is implicated in the pathogenesis of pulmonary fibrosis. Using atomic force microscopy, the authors demonstrate a strong correlation between the cytoskeletal-associated cellular architecture and the mechanical dynamics of alveolar epithelial cells undergoing mesenchymal transition.

N terminus is key to the dominant negative suppression of Ca(V)2 calcium channels: implications for episodic ataxia type 2.

Expression of the calcium channels Ca(V)2.1 and Ca(V)2.2 is markedly suppressed by co-expression with truncated constructs containing Domain I. This is the basis for the phenomenon of dominant negative suppression observed for many of the episodic ataxia type 2 mutations in Ca(V)2.1 that predict truncated channels. The process of dominant negative suppression has been shown previously to stem from interaction between the full-length and truncated channels and to result in downstream consequences of the unfolded protein response and endoplasmic reticulum-associated protein degradation. We have now identified the specific domain that triggers this effect. For both Ca(V)2.1 and Ca(V)2.2, the minimum construct producing suppression was the cytoplasmic N terminus. Suppression was enhanced by tethering the N terminus to the membrane with a CAAX motif. The 11-amino acid motif (including Arg(52) and Arg(54)) within the N terminus, which we have previously shown to be required for G protein modulation, is also essential for dominant negative suppression. Suppression is prevented by addition of an N-terminal tag (XFP) to the full-length and truncated constructs. We further show that suppression of Ca(V)2.2 currents by the N terminus-CAAX construct is accompanied by a reduction in Ca(V)2.2 protein level, and this is also prevented by mutation of Arg(52) and Arg(54) to Ala in the truncated construct. Taken together, our evidence indicates that both the extreme N terminus and the Arg(52), Arg(54) motif are involved in the processes underlying dominant negative suppression.

Simple but powerful: phenanthroline-based small molecules for cellular imaging and cancer screening.

A two-step synthetic procedure gives highly fluorescent phenanthroline molecular probes. The compounds localize in the endoplasmic reticulum and their potential as bioactive probes was evaluated. The materials are quickly taken up by living cells within 5 min. Preliminary in vitro studies have shown that these compounds are selective to esophageal cancer cells and can be used as selective markers in intracellular cancer diagnostics. The materials show a remarkable cytotoxicity towards cancer cells vs normal as 7-1.

Activation of stress-related signalling pathway in human cells upon SiO2 nanoparticles exposure as an early indicator of cytotoxicity.

BACKGROUND: Nanomaterials such as SiO2 nanoparticles (SiO2NP) are finding increasing applications in the biomedical and biotechnological fields such as disease diagnostics, imaging, drug delivery, food, cosmetics and biosensors development. Thus, a mechanistic and systematic evaluation of the potential biological and toxic effects of SiO2NP becomes crucial in order to assess their complete safe applicability limits. RESULTS: In this study, human monocytic leukemia cell line THP-1 and human alveolar epithelial cell line A549 were exposed to a range of amorphous SiO2NP of various sizes and concentrations (0.01, 0.1 and 0.5 mg/ml). Key biological indicators of cellular functions including cell population density, cellular morphology, membrane permeability, lysosomal mass/pH and activation of transcription factor-2 (ATF-2) were evaluated utilizing quantitative high content screening (HCS) approach and biochemical techniques. Despite the use of extremely high nanoparticle concentrations, our findings showed a low degree of cytotoxicity within the panel of SiO2NP investigated. However, at these concentrations, we observed the onset of stress-related cellular response induced by SiO2NP. Interestingly, cells exposed to alumina-coated SiO2NP showed low level, and in some cases complete absence, of stress response and this was consistent up to the highest dose of 0.5 mg/ml. CONCLUSIONS: The present study demonstrates and highlights the importance of subtle biological changes downstream of primary membrane and endocytosis-associated phenomena resulting from high dose SiO2NP exposure. Increased activation of transcription factors, such as ATF-2, was quantitatively assessed as a function of i) human cell line specific stress-response, ii) SiO2NP size and iii) concentration. Despite the low level of cytotoxicity detected for the amorphous SiO2NP investigated, these findings prompt an in-depth focus for future SiO2NP-cell/tissue investigations based on the combined analysis of more subtle signalling pathways associated with accumulation mechanisms, which is essential for establishing the bio-safety of existing and new nanomaterials.