Evidence for the existence of pathogenicity determinants in the Long Terminal Repeats (LTRs) of the Bovine Leukemia Virus (BLV) genome

peer reviewedaudience: researcherEvidence for the existence of pathogenicity determinants in the Long Terminal Repeats (LTRs) of the Bovine Leukemia Virus (BLV) genome Sabrina M. Rodríguez1*, Karina Trono2, Leandro R. Jones3 1 Molecular and Cellular Epigenetics, Interdisciplinary Cluster for Applied Genoproteomics (GIGA) , University of Liège (ULg), Belgium. 2 Instituto de Virología, CICVyA, Instituto Nacional de Tecnología Agropecuaria INTA-Castelar, CC 25 (1712), Castelar. 3 División de Biología Molecular, Estación de Fotobiología Playa Unión, CC 15, Rawson, Chubut 9103, Argentina. *E-mail: [email protected] The majority of BLV-infected animals are asymptomatic carriers (AL) while about 30% develop a benign persistent lymphocytosis (PL). Fatal lymphosarcoma (LS) occurs in 5% of infected animals. The genetic basis of these diverse outcomes of BLV infection is still unknown. Viral LTRs constitute a genetic determinant of pathogenesis for other retroviruses. However, this possibility has never been tested for BLV. Analyses to test correlation between clinical and genotypic traits across species must be corrected by including the group phylogeny. Otherwise, shared evolutionary history can jeopardize statistical independence. Thus, the influence of BLV LTR genetic variation on the clinical manifestation of the disease was investigated by employing Cladistic and Probabilistic, phylogenetic comparative methods. With this purpose, the 5´LTR region of 40 BLV proviruses from bovines with different clinical presentations (AL, PL, LS) was sequenced. Seven polymorphic positions showing an apparent association with the clinical presentation were identified. A provirus phylogeny was obtained using env gene sequences from 28 of the 40 provirus studied in this work. Both Cladistic and Probabilistic comparative analyses based on the empirical sequence alignment and the provirus phylogeny suggested that positions 41 and 56 might be correlated to the clinical presentation. The probabilistic analysis further indicated an association with the viral pathogenesis for positions 373, 450, 494 and 505, though the corresponding statistical supports were lower in comparison to the supports obtained for positions 41 and 56. These observations indicate that the BLV LTRs might contain pathogenicity determinants

KAP1 targets actively transcribed genomic loci to exert pleomorphic effects on RNA polymerase II activity

KAP1 (KRAB-associated protein 1) is best known as a co-repressor responsible for inducing heterochromatin formation, notably at transposable elements. However, it has also been observed to bind the transcription start site of actively expressed genes. To address this paradox, we characterized the protein interactome of KAP1 in the human K562 erythro-leukaemia cell line. We found that the regulator can associate with a wide range of nucleic acid binding proteins, nucleosome remodellers, chromatin modifiers and other transcription modulators. We further determined that KAP1 is recruited at actively transcribed polymerase II promoters, where its depletion resulted in pleomorphic effects, whether expression of these genes was normally constitutive or inducible, consistent with the breadth of possible KAP1 interactors. This article is part of a discussion meeting issue 'Crossroads between transposons and gene regulation'

hMENA11a contributes to HER3-mediated resistance to PI3K inhibitors in HER2-overexpressing breast cancer cells.

Human Mena (hMENA), an actin regulatory protein of the ENA/VASP family, cooperates with ErbB receptor family signaling in breast cancer. It is overexpressed in high-risk preneoplastic lesions and in primary breast tumors where it correlates with HER2 overexpression and an activated status of AKT and MAPK. The concomitant overexpression of hMENA and HER2 in breast cancer patients is indicative of a worse prognosis. hMENA is expressed along with alternatively expressed isoforms, hMENA11a and hMENAΔv6 with opposite functions. A novel role for the epithelial-associated hMENA11a isoform in sustaining HER3 activation and pro-survival pathways in HER2-overexpressing breast cancer cells has been identified by reverse phase protein array and validated in vivo in a series of breast cancer tissues. As HER3 activation is crucial in mechanisms of cell resistance to PI3K inhibitors, we explored whether hMENA11a is involved in these resistance mechanisms. The specific hMENA11a depletion switched off the HER3-related pathway activated by PI3K inhibitors and impaired the nuclear accumulation of HER3 transcription factor FOXO3a induced by PI3K inhibitors, whereas PI3K inhibitors activated hMENA11a phosphorylation and affected its localization. At the functional level, we found that hMENA11a sustains cell proliferation and survival in response to PI3K inhibitor treatment, whereas hMENA11a silencing increases molecules involved in cancer cell apoptosis. As shown in three-dimensional cultures, hMENA11a contributes to resistance to PI3K inhibition because its depletion drastically reduced cell viability upon treatment with PI3K inhibitor BEZ235. Altogether, these results indicate that hMENA11a in HER2-overexpressing breast cancer cells sustains HER3/AKT axis activation and contributes to HER3-mediated resistance mechanisms to PI3K inhibitors. Thus, hMENA11a expression can be proposed as a marker of HER3 activation and resistance to PI3K inhibition therapies, to select patients who may benefit from these combined targeted treatments. hMENA11a activity could represent a new target for antiproliferative therapies in breast cancer

Developing fiber lasers with Bragg reflectors as deep sea hydrophones

The present paper will discuss the work in progress at the Department of Physics of the University of Pisa in collaboration with the IFAC laboratory of CNR in Florence to develop pressure sensors with outstanding sensitivity in the acoustic and ultrasonic ranges. These devices are based on optically-pumped fiber lasers, where the mirrors are Bragg gratings written into the fiber core

SOSIALISASI PELATIHAN PERPAJAKAN BAGI MASYARAKAT PELAKU UMKM KEC. KEDUNGADEM KAB. BOJONEGORO

Kesadaran pajak oleh masyarakat dan wajib pajak merupakan kunci dari masalah perpajakan. Regulasi pajak oleh masyarakat masih sulit dipahami, apalagi jika mengikuti standar pelaporan. Oleh karena itu, dialaksanakan kegiatan pengabdian kepada masyarakat dengan memberikan pemahaman mengenai regulasi perpajakan yang berlaku di Indonesia. Secara spesifik kegiatan pengabdian ini bertujuan melakukan sosialisasi pelatihan perpajakan atas kendala atau masalah pelaporan pajak oleh masyarakat pelaku UMKM serta meningkatkan kesadaran masyarakat akan pajak. Berdasarkan tujuan yang telah ditetapkan, selanjutnya kegiatan pengabdian kepada masyarakat dilaksanakan dengan metode sosialisasi (in- service training). Pengabdian Masyarakat ini dilaksanakan bulan Agustus 2023 bertempat di Kantor Kepala Desa, Krajan, Tumbrasanom, Kec. Kedungadem, Kabupaten Bojonegoro, Jawa Timur. Peserta pelatihan adalah masyarakat pelaku Usaha Mikro Kecil dan Menengah (UMKM). Hasil sosialisasi bisa memberikan pemahaman tentang pajak serta mendorong peserta agar taat pada peraturan dan ketentuan perpajakan di Indonesia dengan efektif. Hasil pelatihan membuktikan bahwa pengabdian ini meningkatkan pemahaman dalam regulasi perpajakan dan metode perhitungan pajak terutama beberapa Pasal PPh 21 dan 25/29 yang kerap diterapkan dalam pembayaran pajak

In-parallel polar monitoring of chemiluminescence emission anisotropy at the solid-liquid interface by an optical fiber radial array

Chemiluminescence (CL) detection is widely employed in biosensors and miniaturized analytical devices since it offers high detectability and flexible device design (there are no geometry requirements for the measurement cell, except the ability to collect the largest fraction of emitted photons). Although the emission anisotropy phenomenon for an emitting dipole bound to the interface between two media with different refractive index is well known for fluorescence, it is still poorly investigated for CL reactions, in which the excited-state reaction products can diffuse in solution before the photon emission event. In this paper, we propose a simple method for the realtime evaluation of the CL emission anisotropy based on a radial array of optical fibers, embedded in a poly(methyl methacrylate) semicylinder and coupled with a Charge-Coupled Device (CCD) camera through a suitable interface. The polar-time evolutions of the CL emission have been studied for catalyzing enzymes immobilized onto a solid surface (heterogeneous configuration) or free in solution (homogeneous configuration). Evidence of the anisotropy phenomenon is observed, indicating that the lifetime of the excited-state products of the enzyme-catalyzed reactions is shorter than the time required for their diffusion in solution at a distance at which the CL can be considered isotropic. These results open new perspectives in the development of CL-based miniaturized analytical devices

A life-attenuated BLV deletant as a candidate vaccine to inhibit viral transmission in bovine herds

A life-attenuated BLV deletant as a candidate vaccine to inhibit viral transmission in bovine herds Sabrina M. Rodríguez1*†, Gerónimo Gutiérrez2*, Arnaud Florins3, Lucas Vagnoni2, Irene Alvarez2, Nicolas Gillet1, Karina Trono2‡, Luc Willems1,3‡ *‡S.M. Rodríguez / G. Gutiérrez and K.Trono / L. Willems contributed equally to this work. 1 Molecular and Cellular Epigenetics, Interdisciplinary Cluster for Applied Genoproteomics (GIGA), University of Liège (ULg), Liège (4100), Belgium. 2 Instituto de Virología, CICVyA, Instituto Nacional de Tecnología Agropecuaria, (1712), Castelar, Argentina. 3 Molecular and Cellular Biology, Gembloux Agro-Bio Tech, University of Liège (ULg), Gembloux (5030), Belgium †E-mail: [email protected] Bovine Leukemia Virus (BLV) is a major sanitary concern in many countries where the virus is widely disseminated among dairy herds with obvious economic impact. Different control strategies have been implemented worldwide to control BLV infection or eradicate the disease with diverse success. Eradication by culling is not economically sustainable in highly infected regions such as Argentina, US or Japan. Segregation of BLV-infected cattle is expensive due to duplication of facilities. Finally, several candidate vaccines based on recombinant viral proteins were unsuccessful to protect from challenge. Therefore, here we propose a novel strategy aimed to decrease seroprevalence based on the employ of a life-attenuated BLV provirus as a candidate vaccine. The rationale behind this strategy is the deletion of genes required to induce pathogenesis leaving those involved in infectivity, resulting in an attenuated deletant with impaired transmissibility. Preliminary experiments showed that the deletant provirus is infectious and elicits an efficient immune response in sheep (n=3) and in the natural host, bovines (n=9). Lack of spread to sentinels further supports the safety of the vaccine. Based on these promissory results, an ongoing validation program is being performed to evaluate the capacity of the candidate vaccine to protect from wild-type BLV infection in herd conditions (n=105). Infection will be routinely monitored and proviral loads will be determined. The efficiency of the immune response will be evaluated by titration of specific antibodies, cytotoxic lysis efficiency and cytokine profile. Viral expression ex vivo and provirus clonality will be also evaluated. This data will be instrumental for understanding the basic mechanisms undergoing during BLV infection and for elaborating a novel vaccine. We do believe this practical and cost-effective vaccination strategy is the sole economically viable in countries with high prevalence

Fiber laser strain sensor device

We present a fiber laser strain sensor (FLSS) with noise-equivalent sensitivity equal to or better than 80?p?rms?(Hz)?1/2 at very low frequencies, from 100?mHz to several hundreds of hertz. The strain affects the fiber laser emission wavelength, and an imbalanced Mach?Zender interferometer (MZI) converts wavelength variations into phase-amplitude variations. The sensor has been also tested in the time domain by applying sinusoidal strain bursts: the device also shows a good signal-to-noise ratio at the lowest burst frequencies

Gene-specific inhibition of breast carcinoma in BALB-neuT mice by active immunization with rat Neu or human ErbB receptors

Employing the transgenic BALB-neuT mouse tumor model, we explored the in vivo biologic relevance of immunocompetent epitopes shared among the four ErbB receptors. The outcome of neu-mediated tumorigenesis was compared following vaccination with isogeneic normal rat ErbB2/Neu (LTR-Neu) or xenogeneic human ErbB receptors (LTR-EGFR, LTR-ErbB2, LTR-ErbB3 and LTR-ErbB4), each recombinantly expressed in an NIH3T3 murine cell background. Vaccination using rat LTR-Neu at the stage of atypical hyperplasia potently inhibited neu-mediated mammary tumorigenesis. Moreover, all human ErbB receptors specifically interfered with tumor development in BALB-neuT mice. Relative increase in tumor-free survival and reduction in tumor incidence corresponded to structural similarity shared with the etiologic neu oncogene, as rat orthologue LTR-Neu proved most effective followed by the human homologue LTR-ErbB2 and the other three human ErbB receptors. Vaccination resulted in high titer specific serum antibodies, whose tumor-inhibitory effect correlated with cross-reactivity to purified rat Neu extracellular domain in vitro. Furthermore, a T cell response specific for peptide epitopes of rat Neu was elicited in spleen cells of mice immunized with LTR-Neu and was remotely detectable for discrete peptides upon vaccination with LTR-ErbB2 and LTR-EGFR. The most pronounced tumor inhibition by LTR-Neu vaccination was associated with leukocyte infiltrate and tumor necrosis in vivo, while immune sera specifically induced cytotoxicity and apoptosis of BALB-neuT tumor cells in vitro. Our findings indicated that targeted inhibition of neu oncogene-mediated mammary carcinogenesis is conditional upon the immunization schedule and discrete immunogenic epitopes shared to a variable extent by different ErbB receptors