Teachers' emotional well-being during the SARS-CoV-2 pandemic with long school closures: a large-scale cross-sectional survey in Northern Italy.

OBJECTIVES: This study aimed to evaluate the magnitude of emotional burden on teaching staff during the SARS-CoV-2 pandemic in a significantly impacted region. In addition, the correlates of emotional burden were analysed to enable the design of targeted interventions. STUDY DESIGN: This study was a cross-sectional survey. METHODS: An electronic survey was administered to the teaching staff at public schools and kindergartens in a specific geographical area. Cross-sectional assessments of pandemic-specific variables were performed using the Pandemic Fatigue Scale, the Depression Anxiety Stress Scale (DASS)-21, the Satisfaction with Life Scale, and the Resilient Coping Scale. DASS-21 results were compared with results from a parallel survey that was representative of the local general population. RESULTS: In total, 3251 teaching staff members participated in the survey. Teachers showed a higher emotional burden for depression, anxiety and stress than the general population during the pandemic. According to a linear regression model, this burden is correlated with the language in which the questionnaires were answered, mistrust towards institutions, specific SARS-CoV-2 anxiety, past infection with SARS-CoV-2, avoidance of information about the pandemic and pandemic fatigue; emotional burden was negatively correlated with measures for life satisfaction, resilience and team atmosphere. Some independent variables were shown to contribute differentially to the variance of depression, anxiety or stress. CONCLUSIONS: Emotional distress during the pandemic among teachers is higher than in the general population and correlates with variables that could, at least in principle, be targeted for specific interventions.

Ethylcellulose nanoparticles as a new "in vitro" transfection tool for antisense oligonucleotide delivery.

Oil-in-water nano-emulsions have been obtained in the HEPES 20 mM buffer solution / [Alkylamidoammonium:Kolliphor EL = 1:1] / [6 wt% ethylcellulose in ethyl acetate] system over a wide oil-to-surfactant range and above 35 wt% aqueous component at 25 °C. The nano-emulsion with an oil-to-surfactant ratio of 70/30 and 95 wt% aqueous component was used for nanoparticles preparation. These nanoparticles (mean diameter around 90 nm and zeta potential of +22 mV) were non-toxic to HeLa cells up to a concentration of 3 mM of cationic species. Successful complexation with an antisense phosphorothioate oligonucleotide targeting Renilla luciferase mRNA was achieved at cationic/anionic charge ratios above 16, as confirmed by zeta potential measurements and an electrophoretic mobility shift assay, provided that no Fetal Bovine Serum is present in the cell culture medium. Importantly, Renilla luciferase gene inhibition shows an optimum efficiency (40%) for the cationic/anionic ratio 28, which makes these complexes promising for "in vitro" cell transfection.

Modulating size and surface charge of ethylcellulose nanoparticles through the use of cationic nano-emulsion templates.

Ethylcellulose nano-emulsions have been obtained by the low-energy phase inversion composition method in the Water / [Alkylamidoammonium: Cremophor WO7] / [6% ethylcellulose in ethyl acetate] system at 25 °C. It is shown that nano-emlulsions' composition variables (oil-to-surfactant ratio, cationic: nonionic surfactant ratio and polymer and water content) produce changes in their droplet diameter, surface charge and colloidal stability following defined trends. Nano-emulsions with good stability, droplet diameters between about 120 and 200 nm and surface charge from about 10 to 50 mV have been obtained. Nano-emulsions are further used as templates for nanoparticle dispersions preparation, which show sizes and surface charges typically smaller and similar respectively to their nano-emulsion templates. Cationic: nonionic surfactant ratio has the highest influence on both, size and surface charge, followed by oil-to-surfactant ratio and water content. Interestingly, the positive charge of the nanoparticles can be depleted under diluting conditions in a time-dependent manner.

Low-energy nano-emulsification approach as a simple strategy to prepare positively charged ethylcellulose nanoparticles.

Positively charged ethylcelulose nanoparticles have been obtained from alkylamidoammonium/Span 80 based nano-emulsion templates. Oil-in-water polymeric nano-emulsions form in a broad range of oil-to-surfactant ratios and water contents above 75 wt% by a low-energy method at 25 °C. Nano-emulsions with a water content of 90 wt% showed droplet sizes typically below 300 nm and high positive zeta potential values (∼55 mV). If oleylamine is added to the system, smaller droplet sizes and higher zeta potential values (∼66 mV) are obtained, but the stability of the nano-emulsions decreases. Although these nano-emulsions are destabilized by creaming, the period of stability is large enough to allow nanoparticle preparation by solvent evaporation. Polymeric nanoparticles obtained show a globular core-shell-like morphology, with mean diameters of around 250 nm. The surface charge of the nanoparticles is similar to that of the nano-emulsion template and remains positive after 24 h dialysis, suggesting slow desorption kinetics of the alkylamidoammonium from the nanoparticle surface. These results indicate that the proposed nano-emulsion approach is a good strategy for the preparation of positively charged nanoparticles from nonionic ethylcellulose polymers.

Influence of early-life nutritional stress on songbird memory formation.

In birds, vocal learning enables the production of sexually selected complex songs, dialects and song copy matching. But stressful conditions during development have been shown to affect song production and complexity, mediated by changes in neural development. However, to date, no studies have tested whether early-life stress affects the neural processes underlying vocal learning, in contrast to song production. Here, we hypothesized that developmental stress alters auditory memory formation and neural processing of song stimuli. We experimentally stressed male nestling zebra finches and, in two separate experiments, tested their neural responses to song playbacks as adults, using either immediate early gene (IEG) expression or electrophysiological response. Once adult, nutritionally stressed males exhibited a reduced response to tutor song playback, as demonstrated by reduced expressions of two IEGs (Arc and ZENK) and reduced neuronal response, in both the caudomedial nidopallium (NCM) and mesopallium (CMM). Furthermore, nutritionally stressed males also showed impaired neuronal memory for novel songs heard in adulthood. These findings demonstrate, for the first time, that developmental conditions affect auditory memories that subserve vocal learning. Although the fitness consequences of such memory impairments remain to be determined, this study highlights the lasting impact early-life experiences can have on cognitive abilities.

Suppressor of cytokine signaling-3 is affected in T-cells from tuberculosisTB patients.

T-cells and T-cell-derived cytokines are crucial mediators of protection against Mycobacterium tuberculosis infection, but these factors are insufficient as biomarkers for disease susceptibility. In order to define T-cell molecules involved in tuberculosis (TB), we compared gene expression profiles of T-cells from patients with active TB, healthy donors with latent M. tuberculosis infection (LTBIs) and non-infected healthy donors (NIDs) by microarray analysis. Pathway-focused analyses identified a prevalent subset of candidate genes involved in the Janus kinase (JAK)-signal transducer and activator of transcription signalling pathway, including those encoding suppressor of cytokine signalling (SOCS) molecules, in the subset of protection-associated genes. Differential expression was verified by quantitative PCR analysis for the cytokine-inducible SH2-containing protein (CISH), SOCS3, JAK3, interleukin-2 receptor α-chain (IL2RA), and the proto-oncogene serine/threonine protein kinase (PIM1). Classification analyses revealed that this set of molecules was able to discriminate efficiently between T-cells from TB patients and those from LTBIs, and, notably, to achieve optimal discrimination between LTBIs and NIDs. Further characterization by quantitative PCR revealed highly variable candidate gene expression in CD4(+) and CD8(+) T-cells from TB patients and only minor differences between CD4(+) and CD8(+) T-cell subpopulations. These results point to a role of cytokine receptor signalling regulation in T-cells in susceptibility to TB.

CD26/dipeptidyl peptidase 4-deficiency alters thymic emigration patterns and leukcocyte subsets in F344-rats age-dependently.

As CD26 (dipeptidyl peptidase 4/DPP4) rapidly truncates incretins N-terminally, including glucagon-like peptide-1, DPP4-inhibitors have been developed for treatment of diabetes type 2. To some extent this is surprising, as CD26/DPP4 is also deeply involved in immune regulation. Long-term pharmacological studies are hampered by off-target inhibition of DPP4-homologues. Therefore, we studied the effects of genetic CD26/DPP4-deficiency by investigating blood, spleen and thymus leucocyte subpopulations of wild-type and CD26-deficient F344-rats at different ages. In young animals at 1 and 3 months of age, there were no differences in leucocyte subsets, while in older animals the T cell composition was changed significantly. From the age of 6 months onwards, reduced numbers of recent thymic emigrants and memory T cells, and consequently an increased amount of naive T cells were observed in CD26-deficient rats. In addition, the architecture of the thymus was altered, as observed by a reduced density of lymphocytes in the medulla. Furthermore, the number of proliferating cells in the thymus was decreased in CD26-deficient rats at a higher age. Moreover, CD26-deficiency resulted in markedly reduced numbers of B cells in later life. Additionally, an age- but not CD26-dependent increase of regulatory T cells and a decrease of natural killer cell numbers were detected in the blood and spleen. Our findings indicate an important role of CD26 in maintaining lymphocyte composition, memory T cell generation and thymic emigration patterns during immunosenescence, with possible implications for using DPP4-inhibitors.

T-cell responses against tuberculin and sensitin in children with tuberculosis and non-tuberculosis mycobacterial lymphadenopathy.

Multi-colour flow cytometry was applied to determine T-cell-specific interferon-gamma, interleukin-2 and tumour necrosis factor-alpha expression in children with tuberculosis and non-tuberculosis mycobacterial lymphadenopathy (NTM-L). In vitro stimulation of peripheral blood mononuclear cells with purified protein derivative from Mycobacterium tuberculosis (tuberculin) and M. avium (sensitin) revealed differential recognition of tuberculin and sensitin in both study groups. Ratios of tuberculin-specific and sensitin-specific T-cell proportions in individual patients discriminated between children with tuberculosis or NTM-L. These findings have the potential to improve the differential diagnosis of mycobacterial infections.

Serotonin (5-HT) enhances the activity of amphotericin B against Aspergillus fumigatus in vitro.

We investigated the in vitro synergistic antifungal potential of combining serotonin (5-HT) and sertraline with amphotericin B and itraconazole against clinical isolates of Aspergillus spp. Synergy tests were performed using the chequerboard microdilution method. Activity was measured against Aspergillus fumigatus (n = 7), Aspergillus flavus (n = 3) and Aspergillus terreus (n = 2), and compared with that for Candida albicans and Candida parapsilosis. The fractional inhibitory concentration (FIC) indices ranged between 0.25 and 3 for the various isolates tested. 5-HT was shown to enhance the activity of amphotericin B against Aspergillus spp. Combination studies with 5-HT and itraconazole and with sertraline and itraconazole or amphothericin B showed different activities for the various strains, including synergism (FIC < 1.0), additivity (FIC = 1), and indifference (FIC between 1.0 and 2.0). 5-HT and sertraline showed antagonistic activity (FIC > 2) with amphotericin B and itraconazole against C. parapsilosis.

Seasonal activation and inactivation of song motor memories in wild canaries is not reflected in neuroanatomical changes of forebrain song areas.

Seasonal, testosterone-dependent changes in sexual behaviors are common in male vertebrates. In songbirds such seasonal changes occur in a learned behavior--singing. Domesticated male canaries (Serinus canaria) appear to lose song units (syllables) after the breeding season and learn new ones until the next breeding season. Here we demonstrate in a longitudinal field study of individual, free-living nondomesticated (wild) canaries (S. canaria) a different mode of seasonal behavioral plasticity, seasonal activation, and inactivation of auditory-motor memories. The song repertoire composition of wild canaries changes seasonally: about 25% of the syllables are sung seasonally; the remainder occur year-round, despite seasonal changes in the temporal patterns of song. In the breeding season, males sing an increased number of fast frequency-modulated syllables, which are sexually attractive for females, in correlation with seasonally increased testosterone levels. About 50% of the syllables that were lost after one breeding season reappear in the following breeding season. Furthermore, some identical syllable sequences are reactivated on an annual basis. The seasonal plasticity in vocal behavior occurred despite the gross anatomical and ultrastructural stability of the forebrain song control areas HVc and RA that are involved in syllable motor control.

Nitric oxide synthase is induced in sporulation of Physarum polycephalum.

The myxomycete Physarum polycephalum expresses a calcium-independent nitric oxide (NO) synthase (NOS) resembling the inducible NOS isoenzyme in mammals. We have now cloned and sequenced this, the first nonanimal NOS to be identified, showing that it shares < 39% amino acid identity with known NOSs but contains conserved binding motifs for all NOS cofactors. It lacks the sequence insert responsible for calcium dependence in the calcium-dependent NOS isoenzymes. NOS expression was strongly up-regulated in Physarum macroplasmodia during the 5-day starvation period needed to induce sporulation competence. Induction of both NOS and sporulation competence were inhibited by glucose, a growth signal and known repressor of sporulation, and by L-N6-(1-iminoethyl)-lysine (NIL), an inhibitor of inducible NOS. Sporulation, which is triggered after the starvation period by light exposure, was also prevented by 1H-[1,2,4]oxadiazolo-[4,3-a]quinoxalin-1-one (ODQ), an inhibitor of NO-sensitive guanylate cyclase. In addition, also expression of lig1, a sporulation-specific gene, was strongly attenuated by NIL or ODQ. 8-Bromo-cGMP, added 2 h before the light exposure, restored the capacity of NIL-treated macroplasmodia to express lig1 and to sporulate. This indicates that the second messenger used for NO signaling in sporulation of Physarum is cGMP and links this signaling pathway to expression of lig1.

[Not Available]. / Gewebsspezifische Veränderungen bei geschlechtsreifen Sauen nach Aufnahme von Zearalenon-hältigem Futter.

Four groups of 16 cyclic sows were fed with 50, 100, 500 or 1000 µg zearalenone per kg feed for 10 days. Afterwards the genital organs were examined routine histologically and lectinhistochemically. Alterations of the lectin binding pattern of the glandular and surface epithelial cells was seen in all four groups, while the routine histology often showed normal results by the two lower concentrations. Macroscopically detectable organ alterations were seen only in the two groups with the highest concentration steps, but not constantly. Clinical symptoms of hyperestrogenismen were very seldom seen.

Exacerbation of rheumatoid arthritis after termination of chemotherapy for breast carcinoma.

Three women with breast carcinoma were treated with combination chemotherapy, including cyclophosphamide, 5-fluorouracil, and methotrexate, after mastectomy. Within two months of termination of chemotherapy, all 3 patients developed florid synovitis. Two patients had prior clinical manifestations consistent with rheumatoid arthritis; one patient had no antecedent history of arthritis. We suggest that this presentation may represent exacerbation of mild or subclinical rheumatoid arthritis as a consequence of withdrawal of methotrexate therapy.

Predictors of recurrence for patients with small (one centimeter or less) localized breast cancer (T1a,b N0 M0).

BACKGROUND: The frequency of small (< or = 1 cm) axillary lymph node negative invasive breast cancers (T1a,b N0 M0) is increasing because of wider implementation of breast cancer screening. Identification of prognostic factors for these patients has been based largely on retrospective pathology review. The authors analyzed histologic factors recorded in the original pathology reports to determine predictors of recurrence for patients with T1a,b N0 M0 breast cancer. METHODS: Two hundred eighteen patients were studied. Potential prognostic factors including measured millimeter tumor size in three dimensions, histologic grade, nuclear grade, and presence or absence of lymphatic vessel invasion were documented prospectively in routine surgical pathology reports of a large community (nonuniversity based) hospital. Follow-up was performed annually by the tumor registry. RESULTS: With a median follow-up of 6.9 years (range, 3-15.8 years), overall recurrence free survival was 93%. Poor nuclear grade (hazard ratio, 5.8; 95% confidence interval, 1.70-19.82; P = 0.004) and lymphatic vessel invasion (hazard ratio, 4.6; 95% confidence interval, 1.34-15.61; P = 0.01) were independent predictors of recurrence. Only 10% of patients had cancers with both poor nuclear grade and lymphatic vessel invasion and their 67% 7-year recurrence free survival (RFS) rate was significantly lower than the 92% RFS rate observed for patients with one of these two factors (P = 0.007) and the 99% RFS for patients with neither poor risk factor (P = 0.0001). CONCLUSIONS: The combination of poor nuclear grade and lymphatic vessel invasion identifies a very small subset (10%) of patients with T1a,b N0 M0 breast cancer with a significant relapse risk that warrants consideration of adjuvant systemic therapy. However, the majority of patients with T1a,b N0 M0 breast cancer have an exceptionally good prognosis.

A randomized trial of etoposide + cisplatin versus vinblastine + bleomycin + cisplatin + cyclophosphamide + dactinomycin in patients with good-prognosis germ cell tumors.

Standard chemotherapy for disseminated germ cell tumors (GCT) cures most patients but causes considerable acute toxicity, including treatment-related death due to septicemia during neutropenia and pulmonary fibrosis. In addition, chronic and delayed toxicities, particularly Raynaud's phenomenon, have been reported in 6% to 37% of treated patients. In an attempt to minimize the acute and chronic effects of treatment which are related primarily to vinblastine and bleomycin, a randomized trial comparing the efficacy and toxicity of vinblastine + bleomycin + cisplatin + cyclophosphamide + dactinomycin (VAB-6) and etoposide + cisplatin (EP) was conducted on 164 eligible patients with good-prognosis GCT. Seventy-nine of 82 (96%) patients receiving VAB-6 and 76/82 (93%) receiving EP achieved a complete remission (CR) with or without adjunctive surgery. Similar proportions of patients in both arms were found at surgery to have necrosis/fibrosis or mature teratoma. With a median follow-up of 24.4 months in the VAB-6 arm and 25.9 months in the EP arm, the total, relapse-free, and event-free survival distributions were similar in the two arms. Patients receiving EP experienced less emesis (P = .05), higher nadir WBC (P = .06) and platelet counts (P = .01), less magnesium wasting (P = .0001), less mucositis (P = .09), and no pulmonary toxicity. No treatment-related mortality was observed. EP is an efficacious and less toxic regimen and is recommended for good-prognosis patients with disseminated GCT.

Abnormal colony formation and prostaglandin E responsiveness of myeloid progenitor cells in patients cured of germ cell neoplasms after combination chemotherapy.

Myelopoiesis was evaluated in 16 patients in complete remission for a minimum of 9 months after treatment with cisplatin-based combination chemotherapy for metastatic germ cell tumors. None had overt hematopoietic abnormalities. Bone marrow aspirates were obtained for routine morphologic evaluation in seven patients, and myeloid precursor cells were studied for both colony/cluster formation and sensitivity to prostaglandin E-(PGE) mediated growth inhibition in 13 patients. Routine stained marrow smears appeared normal. Six of 13 patients demonstrated abnormal colony/cluster formation. Four patients had no colony formation at all. Ten of 12 patients showed decreased sensitivity of granulocyte-macrophage colony-forming cells (CFU-GM) to PGE inhibition. The studies suggest that defects in myelopoiesis are detectable in patients treated for germ cell tumors with combination chemotherapy. The clinical significance of these findings requires long-term follow-up and surveillance for overt hematopoietic abnormalities in survivors of testicular cancer.

Altered renin and aldosterone excretion in patients treated for metastatic germ cell tumours.

Twenty-four patients with metastatic germ cell tumours were studied for abnormalities in the renin-aldosterone axis and for persistent abnormalities of renal function 9+ to 54+ months following completion of cisplatin based chemotherapy. Increased plasma renin activity and aldosterone were identified in fourteen of nineteen (79%) patients. The mean serum magnesium was subnormal. Statistically lower serum phosphorus levels, and higher urea and creatinine levels were also observed. No patients was hypertensive or on diuretics at the time of study. Since vascular toxicity has been reported after cisplatin based chemotherapy and hypomagnesaemia and increased plasma renin activity have been linked to cardiovascular events, these data imply that germ cell tumour patients treated with cisplatin based chemotherapy should be carefully observed for delayed cardiovascular toxicity.

Increased plasma renin and aldosterone in patients treated with cisplatin-based chemotherapy for metastatic germ-cell tumors.

Twenty-four normotensive males in complete remission (CR) for 9+ to 54+ months after cisplatin-based chemotherapy for metastatic germ-cell tumors were evaluated for evidence of alterations in the renin-aldosterone axis and renal function. Abnormally high ambulatory plasma renin activity was seen in 14 of 19 patients with 24-hour urine sodium excretion greater than 50 mEq. This was correlated with elevated ambulatory plasma aldosterone (P = .009) and 24-hour urinary aldosterone excretion (P = .01). The mean serum magnesium value (1.34 +/- .05 mEq/L) was subnormal. Therapy resulted in an increase in serum creatinine during treatment (P less than .0001), an increase in BUN (P less than .01), and decrease in serum phosphorus (P less than .001). The relationship between the alterations in the renin-aldosterone axis and abnormal renal tubular function remains to be determined. In view of reports of cardiovascular toxicity after treatment for germ-cell tumors, and evidence individually linking both magnesium deficiency and increased plasma renin activity (PRA) to cardiovascular consequences, these abnormalities in renin and magnesium metabolism suggest that patients treated with cisplatin-based chemotherapy should be carefully observed for the development of delayed cardiovascular toxicities.