Percutaneous Intramedullary Screw Fixation of Distal Fibula Fractures: A Case Series and Systematic Review

The current reference standard for unstable ankle fractures is open reduction and internal fixation using a plate and lag screws. This approach requires extensive dissection and wound complications are not uncommon. The use of intramedullary screw fixation might overcome these issues. The aim of our study was to provide an overview of the published data regarding intramedullary screw fixation of fibula fractures combined with a small consecutive case series. We performed a search of published studies to identify the studies in which fibula fractures were treated with percutaneous intramedullary screw fixation. Additionally, all consecutive patients treated for an unstable ankle fracture in a level 1 trauma center using an intramedullary screw were retrospectively included. The literature search identified 6 studies with a total of 180 patients. Wound infection was seen in 1 patient (0.6%), anatomic reduction was achieved in 168 patients (93.3%), and a loss of reduction was seen in 2 patients (1.1%). Implant removal was deemed necessary in 3 patients (1.7%) and nonunion was seen is 2 patients (1.1%). A total of 11 patients, in whom no wound complications occurred, were included in our study. The follow-up duration was a minimum of 12 months. A secondary dislocation was seen in 1 patient, and delayed union was observed after 7.5 months in 1 other patient. In conclusion, intramedullary screw fixation is a safe and adequate method to use for fibula fractures, with a low risk of wound complications. Additional research regarding functional outcome is warranted. (C) 2017 by the American College of Foot and Ankle Surgeons. All rights reserve

DYRK1A Controls HIV-1 Replication at a Transcriptional Level in an NFAT Dependent Manner

Transcription of the HIV-1 provirus is regulated by both viral and host proteins and is very important in the context of viral latency. In latently infected cells, viral gene expression is inhibited as a result of the sequestration of host transcription factors and epigenetic modifications. In our present study we analyzed the effect of host factor dual specificity tyrosine-phosphorylation-regulated kinase 1A (DYRK1A) on HIV-1 replication. We show that DYRK1A controls HIV-1 replication by regulating provirus transcription. Downregulation or inhibition of DYRK1A increased LTR-driven transcription and viral replication in cell lines and primary PBMC. Furthermore, inhibition of DYRK1A resulted in reactivation of latent HIV-1 provirus to a similar extent as two commonly used broad-spectrum HDAC inhibitors. We observed that DYRK1A regulates HIV-1 transcription via the Nuclear Factor of Activated T-cells (NFAT) by promoting its translocation from the nucleus to the cytoplasm. Therefore, inhibition of DYRK1A results in increased nuclear levels of NFAT and increased NFAT binding to the viral LTR and thus increasing viral transcription. Our data indicate that host factor DYRK1A plays a role in the regulation of viral transcription and latency. Therefore, DYRK1A might be an attractive candidate for therapeutic strategies targeting the viral reservoi

The effect of DYRK1A inhibition and downregulation is mediated via NFAT.

<p>(A) To analyze the effect of DYRK1A inhibition on the amount of NFAT or NF-kB bound to the viral LTR, a ChIP-qPCR analysis was performed in TZM-bl cells treated with either 240 μM of INDY, the DMSO control, or 12.5 ng/ml TNFα. Sheared DNA was immunoprecipitated with either control IgG, anti-NFAT, or anti-NF-κB antibodies and levels of bound LTR DNA were analyzed by qPCR. (B) The effect of DYRK1A downregulation on LTR driven transcription in the presence of 10μM NF-kB inhibitor BAY or 300 ng/ml NFAT inhibitor FK506 (C) was analyzed by co-transfection of HEK293T cells in 96-wells plates with 5 ng of LTR-luciferase reporter construct and 12.5 ng, 25 ng or 50 ng of shDYRK1A or the shControl vector. Luciferase activity was analyzed 48-hours post-transfection as a measure for LTR activity and expressed relative to the shControl. Data is shown as mean and SD of three independent experiments. (D) Nuclear localization of NFAT was studied in TZM-bl cells cultured for 24 hours in the absence or presence of 240 μM INDY. Subsequently, cells were stained with Hoechst and anti-NFAT and analysed by confocal fluorescent microscopy. Results are representative of at least two independent experiments. Significance was determined with an unpaired student’s T test. *p<0.05, **p<0.01, ***p<0.001.</p

The effect of DYRK1A inhibition on reactivation of HIV-1 LTRs.

<p>(A) The effect of DYRK1A inhibition on reactivation of silent HIV-1 provirus was studied in TZM-bl cells. TZM-bl were treated with either 48 μM, 120 μM or 240 μM of INDY or the DMSO control and 24-hours later LTR-driven luciferase activity was determined as a measure for viral reactivation. Results are expressed relative to the DMSO control. Data is shown as mean and SD of three independent experiments. (B-E) J-Lat cells were treated for twenty four hours with 24, μM, 48 μM, 120 μM or 240 μM of INDY or 12.5 ng/ml TNFα as a positive control. Subsequently the percentage of GFP expressing cells and the mean fluorescent intensity was determined by FACS. Results are expressed relative to the control). Data is shown as mean and SD of two independent experiments. (F) The effect of DYRK1A inhibition on the reactivation of silent HIV-1 provirus was compared to reactivation by two HDAC inhibitors sodium butyrate and TSA. TZM-bl were treated with either 120 μM or 240 μM of INDY, 5 mM or 10 mM of Sodium butyrate, or 9 nM or 18 nM of TSA or the appropriate vehicle control. Twenty-four hours later, LTR-driven luciferase activity was determined and expressed relative to vehicle control. Data is shown as mean and SD of three independent experiments. Significance was determined with an unpaired student’s T test. *p<0.05, **p<0.01, ***p<0.001.</p

Identification of Liver and Plasma microRNAs in Chronic Hepatitis B Virus infection

Background and Aims: With current standard of care a functional cure for Chronic Hepatitis B (CHB) is only achieved in 1-3% of patients and therefore novel therapies are needed. Disease activity during CHB can be determined by a broad range of virological biomarkers, however these biomarkers are also targets for novel treatment strategies. The aim of this study was to identify novel miRNAs that are differentially expressed in plasma and liver in CHB, and determine whether these miRNAs may serve as biomarkers of disease stage or treatment outcome. Methods: miRNA Next-Generation-Sequencing of plasma and liver samples from CHB patient and controls was performed to identify differentially expressed miRNAs. The identified candidate miRNAs were validated by qPCR in additional plasma and liver samples from two CHB cohorts. Results: Several miRNAs in plasma and liver were found to be differentially expressed between CHB patients and controls. Of the identified miRNAs expression levels of miR-122-5p in plasma were associated with plasma HBsAg, and plasma and liver HBV-DNA levels. Expression levels of miR-223-3p, miR-144-5p and miR-133a-3p in liver were associated with plasma alanine aminotransferase levels. No correlation was observed between miRNA expression levels at baseline and treatment outcome. Conclusions: Limited overlap between plasma and liver miRNAs was found, indicating that plasma miRNAs could be useful as biomarkers for treatment outcome or viral activity during treatment. Whereas liver miRNAs are more likely to be regulated by HBV and could be potential therapeutic targets to control viral activity in liver

A Prospective Five-Year Follow-up After peg-Interferon Plus Nucleotide Analogue Treatment or no Treatment in HBeAg Negative Chronic Hepatitis B Patients

Background: Currently available treatment options for chronic hepatitis B (CHB) are not recommended for HBeAg-negative patients with a low viral load. These patients may however benefit from treatment by achieving a functional cure, defined by HBsAg-loss and undetectable HBV DNA. This study evaluated the long-term effect of combination treatment with peg-interferon-alpha-2a (peg-IFN) and adefovir or tenofovir compared to no treatment in these patients. Methods: HBeAg-negative CHB patients with HBV-DNA levels < 20,000 IU/mL (n = 151) were previously randomised 1:1:1 for peg-IFN 180 μg/week plus either adefovir 10 mg/day or tenofovir 245 mg/day, or no treatment and treated for 48 weeks in an open-label study. In this prospective long-term follow-up study, patients were monitored yearly up to five years after end of treatment (week 308). The primary outcome was sustained HBsAg-loss and secondary outcome the dynamics of HBsAg and HBV-DNA levels over time. Results: Of the 131 followed patients, the HBsAg-status was known for 118 patients after five-year follow-up. HBsAg-loss occurred similarly (P = 0.703) in all arms: 8/43 (18.6%) peg-IFN + adefovir, 4/34 (11.7%) peg-IFN + tenofovir, and 6/41 (14.6%) among the untreated patients. The time to HBsAg-loss did not differ between groups (P = 0.641). Low baseline HBsAg levels and genotype A were independently associated with HBsAg-loss irrespective of allocation. HBsAg and HBV-DNA levels declined similarly during follow-up in all patient groups. Conclusions: This prospective randomised controlled study showed that HBsAg-loss overtime was not influenced by treatment with a combination of nucleotide analogue and Peg-IFN. Low baseline HBsAg levels can predict HBsAg-loss irrespective of treatment allocation