Single-step purification and characterization of Pseudomonas aeruginosa azurin.

Azurin is a small periplasmic blue copper protein found in bacterial strains such as Pseudomonas and Alcaligenes where it facilitates denitrification. Azurin is extensively studied for its ability to mediate electron-transfer processes, but it has also sparked interest of the pharmaceutical community as a potential antimicrobial or anticancer agent. Here we offer a novel approach for expression and single-step purification of azurin in Escherichia coli with high yields and optimal metalation. A fusion tag strategy using an N-terminal GST tag was employed to obtain pure protein without requiring any additional purification steps. After the on-column cleavage by HRV 3C Protease, azurin is collected and additionally incubated with copper sulphate to ensure sufficient metalation. UV-VIS absorption, mass spectroscopy, and circular dichroism analysis all validated the effective production of azurin, appropriate protein folding and the development of an active site with an associated cofactor. MD simulations verified that incorporation of the N-terminal GPLGS segment does not affect azurin structure. In addition, the biological activity of azurin was tested in HeLa cells.

Use of EndoAnchors during index endovascular aortic aneurysm repair in patients with hostile proximal aortic neck anatomy.

PURPOSE: Standard endovascular aortic aneurysm repair (EVAR) is sometimes the only treatment option for patients with hostile aortic neck anatomy, but it carries an increased risk of both early and late procedure-related complications. The aim of this study was to report on single-center experience with the Heli-FX EndoAnchors (Medtronic, Santa Rosa, CA) as an adjunctive procedure to endovascular aneurysm repair (EVAR) for prevention and perioperative treatment of proximal neck complications in patients with hostile neck anatomy.  MATERIALS AND METHODS: A single-centre, retrospective study evaluating 24 consecutive patients treated with EndoAnchors during the index EVAR procedure between November 2018 and August 2021. EndoAnchor implantation was indicated for cases with hostile proximal aortic neck anatomy characterised by the presence of at least one of the following parameters: length of 28 mm, angle of >60°, circumferential thrombus/calcification involving ≥50%, and reverse taper. RESULTS:  Median follow-up period was 22.5 months (IQR 2-31.5 months) with no aneurysm-related death, rupture, or conversion to open surgical repair during the follow-up. The procedural success rate was 100%, with no type Ia endoleak at the completion angiography. A mean of 7 EndoAnchors was used per patient (range 4-12). There were no EndoAnchor fractures and dislocations or stent graft fabric damage due to anchor implants. Twenty-three patients (95.8%) remained free of type Ia endoleak and migration on follow-up imaging. Aneurysm sac regression was observed in 13 patients (54.1%), while in 8 patients (33.3%) the sac remained stable. Sac enlargement was present in 1 patient (4.2%) due to late type Ia endoleak. Two patients were lost to the follow-up immediately after the procedure. Between two groups of patients (sac regression versus failure to regress), the larger initial diameter of the proximal neck was the only significant independent factor associated with a lower possibility of sac regression (p= 0,021). CONCLUSIONS:  The use of EndoAnchors during the index EVAR procedure in cases with challenging aortic neck anatomy with or without perioperative type Ia endoleak was associated with good midterm results and led to sac regression in most of the patients (Tab. 4, Fig. 3, Ref. 31).

A Rare Early-Onset Fatal Complication after Transarterial Chemoembolization: A Case Report and Review of the Literature.

Transarterial chemoembolization (TACE) is a minimally invasive treatment for liver cancer, often employed as a bridging therapy or destination treatment for non-operable cases. This case report discusses an 82-year-old woman with a large hepatocellular carcinoma (HCC) who underwent elective TACE due to the high surgical risk associated with her tumor size. Unexpectedly, the patient experienced liver rupture 20 h post-procedure, leading to acute surgical intervention. Despite successful hemostasis during surgery, the patient succumbed to progressive multi-organ failure. We aimed to search the PubMed database for documented cases of ruptured HCC after TACE. This study highlights risk factors for spontaneous HCC rupture and specific factors associated with TACE-induced rupture. Transarterial embolization (TAE) is currently favored as the treatment method for spontaneous ruptures, while the optimal therapy for TACE-induced ruptures remains unclear. In conclusion, this case underscores the importance of recognizing the rare complication of HCC rupture post-TACE and the need for personalized risk assessment. While TAE emerges as a primary treatment choice, the lack of consensus necessitates further studies to establish evidence-based approaches for managing this uncommon yet life-threatening complication.

SLAM Family Receptors in B Cell Chronic Lymphoproliferative Disorders.

The signaling lymphocytic activation molecule (SLAM) receptor family (SLAMF) consists of nine glycoproteins that belong to the CD2 superfamily of immunoglobulin (Ig) domain-containing molecules. SLAMF receptors modulate the differentiation and activation of a wide range of immune cells. Individual SLAMF receptors are expressed on the surface of hematopoietic stem cells, hematopoietic progenitor cells, B cells, T cells, NK cells, NKT cells, monocytes, macrophages, dendritic cells, neutrophils, and platelets. The expression of SLAMF receptors was studied during normal B cell maturation. Several SLAMF receptors were also detected in cancer cell lines of B-lymphoid origin and in pathological B cells from patients with B cell chronic lymphoproliferative disorders (B-CLPD), the most frequent hematological malignancies in adults. This review summarizes current knowledge on the expression of SLAMF receptors and their adaptor proteins SAP and EAT-2 in B-CLPD. Several SLAMF receptors could be regarded as potential diagnostic and differential diagnostic markers, prognostic factors, and targets for the development of novel drugs for patients with B-CLPD.

Positive matrix factorization of seasonally resolved organic aerosol at three different central European background sites based on nuclear magnetic resonance Aerosolomics data.

Concentration data derived from 1H NMR analysis of the water-soluble organic compounds from fine aerosol (PM2.5) at three Central European background stations, Kosetice, Frýdlant (both in the Czech Republic), and Melpitz (Germany), were used for detailed source apportionment analysis. Two winter and two summer episodes (year 2021) with higher organic concentrations and similar wind directions were selected for NMR analyses. The concentration profiles of 61 water-soluble organic compounds were determined by NMR Aerosolomics and a principal component analysis (PCA) was performed on this dataset. Based on the PCA results, 23 compounds were selected for positive matrix factorization (PMF) analysis in order to identify dominant aerosol sources at rural background sites in Central Europe. Both the PCA and the subsequent PMF analyses clearly distinguished the characteristics of winter and summer aerosol particles. In summer, four factors were identified from PMF and were associated with biogenic aerosol (61-78 %), background aerosol (9-15 %), industrial biomass combustion (7-13 %), and residential heating (5-13 %). In winter, only 3 factors were identified - industrial biomass combustion (33-49 %), residential heating (37-45 %) and a background aerosol (8-30 %). The main difference was observed in the winter season with a stronger contribution of emissions from industrial biomass burning at the Czech stations Kosetice and Frýdlant (47-49 %) compared to the Melpitz station (33 %). However, in general, there were negligible differences in identified sources between stations in the given seasons, indicating a certain homogeneity in PM2.5 composition within Central Europe at least during the sampling periods.

Impact of autologous stem cell transplantation (ASCT) on progression free survival (PFS) in newly diagnosed multiple myeloma patients (NDMM) with high risk cytogenetic abnormalities.

OBJECTIVES: ASCT has been considered the standard of care for younger patients with NDMM, however, not all the studies published so far have uniformly demonstrated the complete superiority of ASCT over chemotherapy at standard doses. A systematic review and meta-analysis of randomized studies has shown a significant benefit with single ASCT in terms of prolonged progression-free survival (PFS), but not of overall survival (OS). In our retrospective analysis we investigated the impact of high dose (HD) chemotherapy followed by ASCT in special population of patients with high risk cytogenetic profile on the PFS and treatment outcome. METHODS: Retrospective analysis of NDMM patients eligible for HD chemotherapy followed by upfront ASCT in the era of novel agents, who underwent the ASCT in the Department of hematology and oncohematology LF UPJS and UNLP Kosice in the timeframe of 54 months (from 01/JAN/2019 to 30/JUN/2023). Patients were stratified according to their cytogenetic profile. PFS was defined by the time from ASCT to the disease progression. The OS was defined as the time from the the start of treatment to the death from disease progression. The high risk cytogenetic abnormalities (HRCA) were defined as t(4;14), del(17/17p), t(14;16), t(14;20), nonhyperploidy, gain (1q). RESULTS: Inclusion criteria were met by 65 patients with NDMM who received HD chemotherpy followed by ASCT. We identified 22 (33.8 %) patients with HRCA and 43 (66.2 %) patients with standard cytogenetic risk. During the monitored period we recorded 4 deaths due to disease progression, all of them in the HCRA subgroup. The response was enhanced by the ASCT in both subgroups. The very good partial response (VGPR) increased from 42 % to 46 % and complete remission (CR) increased from 23 % to 45 % after the ASCT. The number of patients achieving only partial response (PR) decreased from 35 % to 9 % after ASCT. In the subgroup of patients with HRCA the median PFS after ASCT was lower compared to the patients with standard cytogenetic risk (17 vs 38 months). The average PFS in both subgroups was 22.9 months. The median OS in both subgroups was not reached, however the only deaths due to disease progression were recorded in the HRCA subgroup. At the time of analysis, 100 % (43) of patients are alive in the standard cytogenetic subgroup versus 72 % (18) of patients in HRCA subgroup. CONCLUSION: HD chemotherapy followed by ASCT remains the standard of care for NDMM eligible for high dose chemotherapy. Our results confirm the benefit of ASCT even in the presence of HRCA. Lower PFS in the HRCA subgroup might indicate the need for more intensive treatment, which may be achieved by tandem ASCT defined as two ASCT performed within a period of no more than six months. Additionally, as three- and four-drug induction therapies are becoming increasingly available and effective, resulting in high minimal residual disease (MRD) negative rates, it is important to continue discussing and further personalizing upfront ASCT to avoid overtreatment and possible toxicities especially in the non-high-risk patient population (Tab. 5, Fig. 2, Ref. 9).