Using a Unique and Long Forgotten Authentic Material in the EFL/ESL Classroom: Poetry

No doubt one of the everlasting concerns of EFL/ESL teachers is to re-examine and improve teaching tools and practices to meet the needs of their students. This article portrays how poetry, a very genuine and authentic text, can enrich students' language experience. My own teaching observations and students' positive feedback, all proved that poetry can enrich emotional response, imaginative power and creativity in the language learners. Thus it increases their engagement and involvement. There are over hundred original ideas for working with poetry in the language classroom. These highly motivational activities and exercises encourage students to express their inner thoughts and feelings in English. No doubt these classroom activities and practices can improve students' language skills. This article provides a sample of a poem worksheet that includes four skills activities: reading, writing, speaking and listening

The dynamic nature of DNA-strand breaks present in differentiating muscle cells and quiescent lymphocytes

AbstractCellular differentiation in a number of eukaryotic systems is associated with changes in the number of DNA-strand breaks and involves the activity of adenosine diphosphoribosyl transferase (ADPRT). DNA-strand breaks are essential for activation of nuclear ADPRT, the activity of which is required for efficient religation of DNA-strand breaks. In this study we demonstrate the dynamic nature of DNA-strand breaks formed in the genome of differentiating avian skeletal muscle cells and quiescent human lymphocytes. Inhibition of ADPRT activity blocks DNA-strand ligation in both cell types and leads to the accumulation of a higher number of strand breaks.DNA-strand breakADP-ribosylationDifferentiationMitogen activatio

Generation of functional CD8+ T Cells by human dendritic cells expressing glypican-3 epitopes

<p>Abstract</p> <p>Background</p> <p>Glypican 3 (GPC-3) is an oncofoetal protein that is expressed in most hepatocellular carcinomas (HCC). Since it is a potential target for T cell immunotherapy, we investigated the generation of functional, GPC-3 specific T cells from peripheral blood mononuclear cells (PBMC).</p> <p>Methods</p> <p>Dendritic cells (DC) were derived from adherent PBMC cultured at 37°C for 7 days in X-Vivo, 1% autologous plasma, and 800 u/ml GM-CSF plus 500 u/ml IL-4. Immature DC were transfected with 20 μg of <it>in vitro </it>synthesised GPC-3 mRNA by electroporation using the Easy-ject plus system (Equibio, UK) (300 V, 150 μF and 4 ms pulse time), or pulsed with peptide, and subsequently matured with lipopolysaccharide (LPS). Six predicted GPC-3 peptide epitopes were synthesized using standard f-moc technology and tested for their binding affinity to HLA-A2.1 molecules using the cell line T2.</p> <p>Results</p> <p>DC transfected with GPC-3 mRNA but not control DC demonstrated strong intracellular staining for GPC-3 and <it>in vitro </it>generated interferon-gamma expressing T cells from autologous PBMC harvested from normal subjects. One peptide, GPC-3_522-530FLAELAYDL, fulfilled our criteria as a naturally processed, HLA-A2-restricted cytotoxic T lymphocyte (CTL) epitope: i) it showed high affinity binding to HLA-A2, in T2 cell binding assay; ii) it was generated by the MHC class I processing pathway in DC transfected with GPC-3 mRNA, and iii) HLA-A2 positive DC loaded with the peptide stimulated proliferation in autologous T cells and generated CTL that lysed HLA-A2 and GPC-3 positive target cells.</p> <p>Conclusions</p> <p>These findings demonstrate that electroporation of GPC-3 mRNA is an efficient method to load human monocyte-derived DC with antigen because <it>in vitro </it>they generated GPC-3-reactive T cells that were functional, as shown by interferon-gamma production. Furthermore, this study identified a novel naturally processed, HLA-A2-restricted CTL epitope, GPC-3_522-530FLAELAYDL, which can be used to monitor HLA-A2-restricted CTL responses in patients with HCC. Further studies are required to investigate whether anti-GPC-3 immunotherapy has a role in the treatment of GPC-3 dependent tumours, such as HCC.</p

Comparative evaluation of the effect of Er:YAG laser and low level laser irradiation combined with CPP-ACPF cream on treatment of enamel caries

Objectives: This study investigated the effectiveness of low power red and infrared lasers and that of Er:YAG laser, in association with CPP-ACPF cream, on remineralization of white spot lesions. Study Design: Fifty intact premolars were immersed in a demineralization solution for 10 weeks to induce caries like lesions and then were divided into five groups. In group 1, the teeth were covered with a CPP-ACPF cream for 3 minutes and then irradiated with a low power red laser (660 nm, 200 mW) for 1 minute through the cream. In group 2, the treatment was the same as that in group 1, but an infrared laser (810 nm, 200 mW) was employed. The specimens in group 3 were irradiated with an Er:YAG laser (100 mJ, 10 Hz) combined with CPP-ACPF. In group 4, the CPP-ACPF cream was applied for 4 minutes and group 5 was submitted to neither laser nor CPP-ACPF. The micro Vickers hardness was compared at 20, 60 and 100 ì from the enamel surface among the groups. Results: The highest microhardness was observed in the low power red and Er:YAG laser groups and the lowest one belonged to the CPP-ACPF alone and control groups. However, no significant difference was found in microhardness of the experimental groups at any of the evaluation depths (p>0.05). Conclusion: With the laser parameters used in this study, neither the combined application of Er:YAG laser with CPP-ACPF nor the combination of low power lasers with CPP-ACPF provided a significant increase in remineralization of enamel caries

Triggering of Toll-like receptors in the elderly. A pilot study relevant for vaccination

The impaired ability of the elderly to mount an efficient immune response after exposure to microbes or vaccines represents a major challenge in protection against pathogens in ageing. Recently studies have shown that stimulation of Toll-like receptors (TLRs), using stimulatory ligands, can enhance vaccine efficacy by a number of mechanisms, including the activation of innate immune cells and the consequent production of inflammatory cytokines

COVID-19: Time for precision epidemiology

The global COVID-19 (SARS-CoV2, COVID-19) tsunami caused by SARSCoV2 is inundating and often-overwhelming health care systems in most countries and regions..

Analysis of T and NK cell subsets in Sicilian population from young to supercentenarian: the role of age and gender

Ageing dramatically affects number and function of both innate and adaptive arms of immune system, particularly T cell subsets, contributing to reduced vaccination efficacy, decreased resistance to infections and increased prevalence of cancer in the older people. In the present paper, we analysed the age-related changes in the absolute number of lymphocytes in 214 Sicilian subjects, and in the percentages of T and NK cells in a sub-cohort of donors. We compared these results with the immunophenotype of the oldest living Italian supercentenarian (111 years old). The results were also sorted by gender. The correlation between number/percentage of cells and age in all individuals and, separately, in males and females, was examined using a simple linear regression analysis. We did not record the increase in the rate of inversion of the CD4/CD8 ratio frequently reported as associated with ageing in literature. Our observation was the direct consequence of a flat average trend of CD4+ and CD8+ T cell percentages in ageing donors, even when gender differences were included. Our results also suggest that CD4+ and CD8+ subsets are not affected equally by age comparing females with males, and we speculated that gender may affect the response to CMV infection. The supercentenarian showed a unique immunophenotypic signature as regards the relative percentages of her T cell subsets, with CD4+ and CD8+ T cell percentages and CD4+ na\uefve T cell values in line with those recorded for the octogenarian subjects. This suggests that the supercentenarian has a na\uefve "younger" T cell profile comparable to that of a &gt;80 year old female

Lentiviral Engineered Fibroblasts Expressing Codon-Optimized COL7A1 Restore Anchoring Fibrils in RDEB

Cells therapies, engineered to secrete replacement proteins, are being developed to ameliorate otherwise debilitating diseases. Recessive dystrophic epidermolysis bullosa (RDEB) is caused by defects of type VII collagen, a protein essential for anchoring fibril formation at the dermal-epidermal junction. Whereas allogeneic fibroblasts injected directly into the dermis can mediate transient disease modulation, autologous gene-modified fibroblasts should evade immunological rejection and support sustained delivery of type VII collagen at the dermal-epidermal junction. We demonstrate the feasibility of such an approach using a therapeutic grade, self-inactivating-lentiviral vector, encoding codon-optimized COL7A1, to transduce RDEB fibroblasts under conditions suitable for clinical application. Expression and secretion of type VII collagen was confirmed with transduced cells exhibiting supranormal levels of protein expression, and ex vivo migration of fibroblasts was restored in functional assays. Gene-modified RDEB fibroblasts also deposited type VII collagen at the dermal-epidermal junction of human RDEB skin xenografts placed on NOD-scid IL2Rgammanull recipients, with reconstruction of human epidermal structure and regeneration of anchoring fibrils at the dermal-epidermal junction. Fibroblast-mediated restoration of protein and structural defects in this RDEB model strongly supports proposed therapeutic applications in man

A functional assay for microRNA target identification and validation

MicroRNAs (miRNA) are a class of small RNA molecules that regulate numerous critical cellular processes and bind to partially complementary sequences resulting in down-regulation of their target genes. Due to the incomplete homology of the miRNA to its target site identification of miRNA target genes is difficult and currently based on computational algorithms predicting large numbers of potential targets for a given miRNA. To enable the identification of biologically relevant miRNA targets, we describe a novel functional assay based on a 3′-UTR-enriched library and a positive/negative selection strategy. As proof of principle we have used mir-130a and its validated target MAFB to test this strategy. Identification of MAFB and five additional targets and their subsequent confirmation as mir-130a targets by western blot analysis and knockdown experiments validates this strategy for the functional identification of miRNA targets

A Phase 1 trial of human telomerase reverse transcriptase (hTERT) vaccination combined with therapeutic strategies to control immune-suppressor mechanisms

The presence of inhibitory immune cells and difficulty in generating activated effector T-cells remain obstacles to development of effective cancer vaccines. We designed a vaccine regimen combining human telomerase reverse transcriptase (hTERT) peptides with concomitant therapies targeting regulatory T-cells (Tregs) and cyclooxygenase-2 (COX2)-mediated immunosuppression. This Phase 1 trial combined an hTERT-derived 7-peptide library, selected to ensure presentation by both HLA class-I and class-II in 90% of patients, with oral low-dose cyclophosphamide (to modulate Tregs) and the COX2 inhibitor celecoxib. Adjuvants were Montanide and topical TLR-7 agonist, to optimise antigen presentation. The primary objective was determination of the safety and tolerability of this combination therapy, with anti-cancer activity, immune response and detection of antigen-specific T-cells as additional endpoints. Twenty-nine patients with advanced solid tumours were treated. All were multiply-pretreated, and the majority had either colorectal or prostate cancer. The most common adverse events were injection-site reactions, fatigue and nausea. Median progression-free survival was 9 weeks, with no complete or partial responses, but 24% remained progression-free for ≥6 months. Immunophenotyping showed post-vaccination expansion of CD4+ and CD8+ T-cells with effector phenotypes. The in vitro re-challenge of T-cells with hTERT peptides, TCR sequencing, and TCR similarity index analysis demonstrated the expansion following vaccination of oligoclonal T-cells with specificity for hTERT. However, a population of exhausted PD-1 + cytotoxic T-cells was also expanded in vaccinated patients. This vaccine combination regimen was safe and associated with antigen-specific immunological responses. Clinical activity could be improved in future by combination with anti-PD1 checkpoint inhibition to address the emergence of an exhausted T-cell population