MicroRNA-29 family expression and its relation to antiviral immune response and viro-immunological markers in HIV-1-infected patients

Abstract BACKGROUND: Several in vitro studies suggested the microRNA-29 (miRNA-29) family is involved in regulating HIV-1 and modulating the expression of interleukin (IL)-32, an anti-HIV-1 cytokine. METHODS: To investigate the contribution of the miRNA-29 family to HIV-1 infection in vivo, we compared miRNA-29 expression in PBMC collected from 58 HIV-1-infected patients, naïve for antiretroviral therapy, and 21 gender- and age-matched HIV-1 seronegative healthy donors, using RT-Taqman assays. The relation between miRNA-29 levels and HIV-1 viro-immunological markers and the activation rate of antiviral immune response were also evaluated. In addition, we profiled miRNA-29 expression in CD4+ T lymphocytes and CD14+ monocytes collected from 5 antiretroviral treated HIV-1 infected patients. RESULTS: miRNA-29b levels were higher in HIV-1-infected patients than in the control group (p < 0.001). There were no correlations with either HIV-1 RNA levels or CD4+ T count, whereas a significant correlation was found between miRNA-29-a/c levels and integrated HIV-1 DNA (miRNA-29a: p = 0.009, r = -0.448; miRNA-29c: p = 0.029; r = -0.381). When the HIV-1-infected patients were grouped on the basis of their plasma HIV-1 RNA and CD4+ T cell count, we also found that patients expressing the lowest levels of miRNA-29c showed high viraemia, low CD4+ T cell count and high levels of integrated HIV-1 DNA. Moreover, miRNA-29b levels were correlated with those of IL-32nonα (p = 0.028; r = -0.298). Patients expressing higher levels of miRNA-29b showed lower levels of MxA, an interferon-stimulated gene, also induced by IL-32 (p = 0.006 r = -0.397). Lastly, we found that CD4+ T lymphocytes and CD14+ monocytes shared similar miRNA-29a/b/c expression patterns but the amount of miRNA-29a/b/c, IL-32 isoforms and MxA were highly variable in these two cellular subsets. CONCLUSIONS: The miRNA-29 family could influence the clinical progression of HIV-1 infection, the HIV-1 proviral load and the innate immune response against HIV-1

Biomechanics of anterior cruciate ligament reconstruction using twisted doubled hamstring tendons

We studied the biomechanical properties of a twisted doubled semitendinosus and gracilis graft. We applied an un-axial load in order to reproduce the kinematics of a reconstructed anterior cruciate ligament (ACL). A modified cryo-jaw clamp system was used to minimize soft tissue slippage. The lower grip, after fixation of the free ends of the tendons, was rotated 45degrees, translated 1 cm, and bent 40degrees, simulating a knee sprain. The graft was tested to failure using a servohydraulic machine. The specimen from one knee of seven unembalmed cadavers was assigned to the untwisted (parallel) bundles group, while its pair was assigned to the twisted group. For the parallel bundles group, the mean maximum load was 1709.3+/-581.9 N, for the twisted group 2428.3+/-475.4 N (P<0.05). The mean stiffness was respectively 213.6+/-72.4 N/mm and 310.3+/-97.3 N/mm (P=0.08). Although caution should be used in extrapolating the results to clinical estimates of the strength of hamstring grafts, the results of the present study could justify the use of twisted semitendinosus and gracilis bundles in ACL reconstruction

A modified Cryo-Jaw for in vitro biomechanical testing of tendons

The purpose of this study was to develop a new device, which represents a modification of the Cryo-Jaw described by Riemersa and Schamhardt and modified by Hamner et al., for in vitro biomechanical testing of tendons which allows the lower clamp to move in every direction and thus simulate a pathological dislocation of the knee. Tendons are fixed to the device by freezing the clamped part with dry ice. After fixation of their free ends. the lower clamp was rotated 45, translated 1 em, and angled 40 to simulate a knee sprain, Various configurations of bundles were tested: parallel, twisted, and braided. Tests were performed on 10 paired bovine bifurcated digital extensor tendons and 6 paired human hamstring tendons. Grafts were then tested to failure subjected to impulsive load, using a servohydraulic machine. The highest ultimate load recorded for parallel bundles was 4662 +/- 565.71 N for bovine bifurcated digital extensor tendons, and 3057 +/- 475.44 N for human hamstring tendons. In any case, the tendons ruptured midway, well clear of the frozen part; in no case was slippage of the tendons observed. Thus the device proposed allows one to test what happens to the graft of an ACL reconstructed knee during physiological and pathological movements because it can be easily displaced in every direction

Tissue Inhibitor of Metalloproteinase-3 Regulates Inflammation in Human and Mouse Intestine

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Controlled Swapping of Nanocomposite Surface Wettability by Multilayer Photopolymerization

Single-layered photopolymerized nanocomposite films of polystyrene and TiO2 nanorods change their wetting characteristics from hydrophobic to hydrophilic when deposited on substrates with decreasing hydrophilicity. Interestingly, the addition of a second photopolymerized layer causes a swapping in the wettability, so that the final samples result converted from hydrophobic to hydrophilic or vice versa. The wettability characteristics continue to be swapped as the number of photopolymerized layers increases. In fact, odd-layered samples show the same wetting behavior as single-layered ones, while even-layered samples have the same surface characteristics as double-layered ones. Analytical surface studies demonstrate that all samples, independently of the number of layers, have similar low roughness, and that the wettability swap is due to the different concentration of the nanocomposites constituents on the samples surface. Particularly, the different interactions between the hydrophilic TiO2 nanorods and the underlying layer lead to different amounts of nanorods exposed on the nanocomposites surface. Moreover, due to the unique property of TiO2 to reversibly increase its wettability upon UV irradiation and subsequent storage, the wetting characteristics of the multilayered nanocomposites can be tuned in a reversible manner. In this way, a combination of substrate, number of photopolymerized layers, and external UV light stimulus can be used in order to precisely control the surface wettability properties of nanocomposite films, opening the way to a vast number of potential applications in microfluidics, protein assays, and cell growth

Light-controlled directional liquid drop movement on TiO2 nanorods-based nanocomposite photopatterns.

Patterned polymeric coatings enriched with colloidal TiO(2) nanorods and prepared by photopolymerization are found to exhibit a remarkable increase in their water wettability when irradiated with UV laser light. The effect can be completely reversed using successive storage in vacuum and dark ambient environment. By exploiting the enhancement of the nanocomposites hydrophilicity upon UV irradiation, we prepare wettability gradients along the surfaces by irradiating adjacent surface areas with increasing time. The gradients are carefully designed to achieve directional movement of water drops along them, taking into account the hysteresis effect that opposes the movement as well as the change in the shape of the drop during its motion. The accomplishment of surface paths for liquid flow, along which the hydrophilicity gradually increases, opens the way to a vast number of potential applications in microfluidics

Nucleosomes effectively shield DNA from radiation damage in living cells

Abstract Eukaryotic DNA is organized in nucleosomes, which package DNA and regulate its accessibility to transcription, replication, recombination and repair. Here, we show that in living cells nucleosomes protect DNA from high-energy radiation and reactive oxygen species. We combined sequence-based methods (ATAC-seq and BLISS) to determine the position of both nucleosomes and double strand breaks (DSBs) in the genome of nucleosome-rich malignant mesothelioma cells, and of the same cells partially depleted of nucleosomes. The results were replicated in the human MCF-7 breast carcinoma cell line. We found that, for each genomic sequence, the probability of DSB formation is directly proportional to the fraction of time it is nucleosome-free; DSBs accumulate distal from the nucleosome dyad axis. Nucleosome free regions and promoters of actively transcribed genes are more sensitive to DSB formation, and consequently to mutation. We argue that this may be true for a variety of chemical and physical DNA damaging agents

Anti-Survival Effect of SI306 and Its Derivatives on Human Glioblastoma Cells

Glioblastoma (GBM) is the most common adult brain tumor and, although many efforts have been made to find valid therapies, the onset of resistance is the main cause of recurrence. Therefore, it is crucial to identify and target the molecular mediators responsible for GBM malignancy. In this context, the use of Src inhibitors such as SI306 (C1) and its prodrug (C2) showed promising results, suggesting that SI306 could be the lead compound useful to derivate new anti-GBM drugs. Therefore, a new prodrug of SI306 (C3) was synthesized and tested on CAS-1 and U87 human GBM cells by comparing its effect to that of C1 and C2. All compounds were more effective on CAS-1 than U87 cells, while C2 was the most active on both cell lines. Moreover, the anti-survival effect was associated with a reduction in the expression of epidermal growth factor receptor (EGFR)WT and EGFR-vIII in U87 and CAS-1 cells, respectively. Collectively, our findings demonstrate that all tested compounds are able to counteract GBM survival, further supporting the role of SI306 as progenitor of promising new drugs to treat malignant GBM

Pokušaji lociranja i uzorkovanja velike bijele psine, Carcharodon carcharias (Lamniformes: Lamnidae), uz talijansku obalu u Sredozemnom moru

Described in the present report are documented attempts to tag and sample the white shark, Carcharodon carcharias (Linnaeus, 1758), along Italian coasts in the Mediterranean Sea, which took place near the Lampedusa Island in the lower Tyrrhenian Channel of Sicily, off the coast of Rimini in the Adriatic Sea, and of Civitavecchia in the central Tyrrhenian Sea. The project, activated in 2015, was aimed at tagging and sampling specimens of the white shark in order to collect data useful to implement conservation strategies in the Mediterranean Sea. Despite four tagging attempts made in 2017, 2018, 2021, and 2022 with 288 total hours of baiting activity and the use of 1030 kg of chum, no white sharks or any other shark species were sighted.U radu su dokumentirani pokušaji označavanja i uzorkovanja velike bijele psine Carcharodon carcharias (Linnaeus, 1758) uz talijansku obalu u Sredozemnom moru. Istraživanje je obavljeno na području oko otoka Lampeduze, u sicilijanskom kanalu u južnom Tirenskom moru, ispred Riminija u Jadranskom moru te kod Civitavecchije u centralnom Tirenskom moru. Projekt, koji je započeo 2015. godine, bio je usmjeren na označavanje i uzorkovanje primjeraka velike bijele psine s ciljem prikupljanja podataka važnih za implementaciju mjera za očuvanje populacije ove vrste u Sredozemnom moru. Unatoč četiri pokušaja označavanja obavljena 2017., 2018., 2021. i 2022. godine, u ukupnom trajanju od 288 sati primamljivanja uz pomoć mamca te 1030 kg mamca, ni jedan primjerak velike bijele psine, ni drugih vrsta morskih pasa, nije primijećen