Raman microscopy reveals how cell inflammation activates glucose and lipid metabolism.

Metabolism of endothelial cells (ECs) depends on the availability of the energy substrates. Since the endothelium is the first line of defence against inflammation in the cardiovascular system and its dysfunction can lead to the development of cardiovascular diseases, it is important to understand how glucose metabolism changes during inflammation. In this work, glucose uptake was studied in human microvascular endothelial cells (HMEC-1) in high glucose (HG), and additionally in an inflammatory state, using Raman imaging. HG state was induced by incubation of ECs with a deuterated glucose analogue, while the EC inflammation was caused by TNF-α pre-treatment. Spontaneous and stimulated Raman scattering spectroscopy provided comprehensive information on biochemical changes, including lipids and the extent of unsaturation induced by excess glucose in ECs., induced by excess glucose in ECs. In this work, we indicated spectroscopic markers of metabolic changes in ECs as a strong increase in the ratio of the intensity of lipids / (proteins + lipids) bands and an increase in the level of lipid unsaturation and mitochondrial changes. Inflamed ECs treated with HG, revealed enhanced glucose uptake, and intensified lipid production i.a. of unsaturated lipids. Additionally, increased cytochrome c signal in the mitochondrial region indicated higher mitochondrial activity and biogenesis. Raman spectroscopy is a powerful method for determining the metabolic markers of ED which will better inform understanding of disease onset, development, and treatment.

What is the ability of inflamed endothelium to uptake exogenous saturated fatty acids? A proof-of-concept study using spontaneous Raman, SRS and CARS microscopy.

Endothelial cells (EC) in vivo buffer and regulate the transfer of plasma fatty acid (FA) to the underlying tissues. We hypothesize that inflammation could alter the functionality of the EC, i.e., their capacity and uptake of different FA. The aim of this work is to verify the functionality of inflamed cells by analyzing their ability to uptake and accumulate exogenous saturated FA. Control and inflammatory human microvascular endothelial cells stimulated in vitro with two deuterium-labeled saturated FA (D-FA), i.e., palmitic (D31-PA) and myristic (D27-MA) acids. Cells were measured both by spontaneous and stimulated Raman imaging to extract detailed information about uptaken FA, whereas coherent anti-Stokes Raman scattering and fluorescence imaging showed the global content of FA in cells. Additionally, we employed atomic force microscopy to obtain a morphological image of the cells. The results indicate that the uptake of D-FA in inflamed cells is dependent on their concentration and type. Cells accumulated D-FA when treated with a low concentration, and the effect was more pronounced for D27-MA, in normal cells, but even more so, in inflamed cells. In the case of D31-PA, a slightly increased uptake was observed for inflamed cells when administered at higher concentration. The results provide a better understanding of the EC inflammation and indicate the impact of the pathological state of the EC on their capacity to buffer fat. All the microscopic methods used showed complementarity in the analysis of FA uptake by EC, but each method recognized this process from a different perspective.

EdU sensing: The Raman way of following endothelial cell proliferation in vitro and ex vivo.

Endothelial cells line the lumen of all vessels in the body and maintain vascular homeostasis. In particular, endothelial cell regeneration in response to insult sustain functional endothelial layer. EdU (5-ethynyl-2'-deoxyuridine) is an alkyne-tagged proliferation probe that incorporates into newly synthesized DNA and is used for fluorescence imaging of cell proliferation with the use of "click chemistry" reaction with a fluorescent azide. Here, we utilized EdU as a click-free Raman probe for tracking endothelial cell proliferation. Raman imaging of EdU was performed in live endothelial cells, showing an advantage over fluorescence imaging of EdU, as this technique did not require sample fixation and permeabilization. To validate Raman-based imaging of EdU to study endothelial cell proliferation, we showed that when endothelial cells were treated with cycloheximide or doxorubicin to impair the proliferation of endothelial cells, the Raman-based signal of EdU was diminished. Furthermore, endothelial cells proliferation detected using EdU-labelled Raman imaging was compared with fluorescence imaging. Finally, the method of Raman-based EdU imaging was used in the isolated murine aorta ex vivo. Altogether, our results show that Raman-based imaging of EdU provides a novel alternative for fluorescence-based assay to assess endothelial proliferation and regeneration.

NMR and MD Analysis of the Bonding Interaction of Vancomycin with Muramyl Pentapeptide.

The article describes an NMR spectroscopy study of interactions between vancomycin and a muramyl pentapeptide in two complexes: vancomycin and a native muramyl pentapeptide ended with D-alanine (MPP-D-Ala), and vancomycin and a modified muramyl pentapeptide ended with D-serine (MPP-D-Ser). The measurements were made in a 9:1 mixture of H2O and D2O. The obtained results confirmed the presence of hydrogen bonds previously described in the literature. At the same time, thanks to the pentapeptide model used, we were able to prove the presence of two more hydrogen bonds formed by the side chain amino group of L-lysine and oxygen atoms from the vancomycin carboxyl and amide groups. This type of interaction has not been described before. The existence of these hydrogen bonds was confirmed by the 1H NMR and molecular modeling. The formation of these bonds incurs additional through-space interactions, visible in the NOESY spectrum, between the protons of the L-lysine amino group and a vancomycin-facing hydrogen atom in the benzylic position. The presence of such interactions was also confirmed by molecular dynamics trajectory analysis.

Influence of Trajectory and Dynamics of Vehicle Motion on Signal Patterns in the WIM System.

This paper presents the analyses of the signals recorded by the main sensors of a WIM test station in the cases of abnormal runs (i.e., runs with the changes of trajectory or the dynamics of vehicle motion). The research involved strain gauges which are used for measuring the weight of vehicles, inductive loops, as well as piezoelectric sensors used, inter alia, to detect twin wheels and to determine where a vehicle passes through a station. Since the designers intend the station to be able to implement the direct enforcement function, the selection of runs deviating from the normative ones constitutes an important issue for the assessment of the measurement reliability. The study considered the location of the trajectory of the runs, the dynamics (acceleration/braking) and the trajectory changes. The change in the amplitude and the value of the signal recorded by the strain gauges as a function of the location (position) of the contact between sensor and tires is a noteworthy observation which indicates the need to monitor this parameter in automatic WIM systems. Other tests also demonstrated the influence of the analysed driving parameters on the recorded results. However, by equipping the WIM station with a set of duplicate strain gauges, the measurement errors of the gross weight and axle loads are normally within the accuracy limits of class A(5) stations. Only in the case of accelerating/decelerating, does the error in measuring the load of a single axle reach several per cent.

A new in vitro model applied 90Y microspheres to study the effects of low dose beta radiation on colorectal cancer cell line in various oxygenation conditions.

We propose a new in vitro model to assess the impact of 90Y-microspheres derived low-dose beta radiation on colorectal cancer cell line under various oxygenation conditions that mimic the tumor environment. Cancer cells (HCT116) proliferation was assessed using Alamar Blue (AB) assay after 48, 72, and 96 h. FLUKA code assessed changes in cancer cell populations relative to the absorbed dose. In normoxia, mitochondrial activity measured by Alamar Blue after 48-72 h was significantly correlated with the number of microspheres (48 h: r = 0.87 and 72 h: r = 0.89, p < 0.05) and absorbed dose (48 h: r = 0.87 and 72 h: r = 0.7, p < 0.05). In hypoxia, the coefficients were r = 0.43 for both the number of spheres and absorbed dose and r = 0.45, r = 0.47, respectively. Impediment of cancer cell proliferation depended on the absorbed dose. Doses below 70 Gy could reduce colorectal cancer cell proliferation in vitro. Hypoxia induced a higher resistance to radiation than that observed under normoxic conditions. Hypoxia and radiation induced senescence in cultured cells. The new in vitro model is useful for the assessment of 90Y radioembolization effects at the micro-scale.

Intra-aneurysmal pressure changes during stent-assisted coiling.

We aimed to examine aneurysm hemodynamics with intra-saccular pressure measurement, and compare the effects of coiling, stenting and stent-assisted coiling in proximal segments of intracranial circulation. A cohort of 45 patients underwent elective endovascular coil embolization (with or without stent) for intracranial aneurysm at our department. Arterial pressure transducer was used for all measurements. It was attached to proximal end of the microcatheter. Measurements were taken in the parent artery before and after embolization, at the aneurysm dome before embolization, after stent implantation, and after embolization. Stent-assisted coiling was performed with 4 different stents: LVIS and LVIS Jr (Microvention, Tustin, CA, USA), Leo (Balt, Montmorency, France), Barrel VRD (Medtronic/ Covidien, Irvine, CA, USA). Presence of the stent showed significant reverse correlation with intra-aneurysmal pressure-both systolic and diastolic-after its implantation (r = -0.70 and r = -0.75, respectively), which was further supported by correlations with stent cell size-r = 0.72 and r = 0.71, respectively (P<0.05). Stent implantation resulted in significant decrease in diastolic intra-aneurysmal pressure (p = 0.046). Systolic or mean intra-aneurysmal pressure did not differ significantly. Embolization did not significantly change the intra-aneurysmal pressure in matched pairs, regardless of the use of stent (p>0.05). In conclusion, low-profile braided stents show a potential to divert blood flow, there was significant decrease in diastolic pressure after stent placement. Flow-diverting properties were related to stent porosity. Coiling does not significantly change the intra-aneurysmal pressure, regardless of packing density.

Monitoring road traffic participants' exposure to PM10 using a low-cost system.

The paper presents the application of a low-cost system for monitoring the current level of road traffic participants' exposure to PM10 air pollution. The research was carried out from the end of August 2017 to the beginning of October 2017 on the central section of one of the main roads in Bielsko-Biala, Poland. In the analysed period, significant changes in the daily distribution of road traffic both into and out of the city centre were observed. The average travel time depended on the direction of traffic, and the difference between directions being almost 50%. The PM10 urban background concentration was also subject to daily changes, and in the fifth week of observation, it reached a value more than twice as high as in the first week of observation. The maximum level of road traffic participants' exposure was observed at a relatively low urban background PM10 concentration. It was observed that a significant slowdown in traffic in conditions of acceptable urban air quality led to a comparable level of exposure to that of standard traffic conditions and poor urban air quality. It was also found that the slowdown in traffic increased the exposure time of traffic participants travelling towards the city centre by an average of 24% and, for those travelling in the opposite direction, by as much as 50%. In an extreme case of traffic delay, exposure to PM10 concentration in the vicinity of the road was two and a half times as long.

Gallbladder Radiation Protection in SIRT-Quantitative Anatomical Study of Hepatic Vasculature.

Introduction: This study was designed to assess quantitatively a safe position of the microcatheter during the SIRT (Selective Internal Radiation Therapy) procedure, in order to minimize the risk of non-target spheres leaking. Materials and Methods: Retrospective analysis of the distance of the tip of the microcatheter from coiled or patent non-target arteries was measured during 99mTc-MAA work-up procedure. Frequency of extrahepatic leaking during work-up and SIRT procedures was evaluated. Results: There were 85 patients who underwent 98 work-up procedures. There were 64 radioembolizations. There were 44 gastroduodenal, 51 right gastric, and 54 cystic artery embolizations performed. Extrahepatic 99mTc-MAA leaking was observed in 33 cases: 16 to gallbladder, four to a gastric wall, nine to the duodenum, one to the intestinal wall, and three to the abdominal wall. Leak in 99mTc-MAA was also related to the presence of additional arteries (p = 0.009). There were 34 proximal and 31 distal to cystic artery 99mTc-MAA injections resulting in 12 vs. four leaks, respectively (p = 0.039, RR-2.5). Mean distance of the tip of the microcatheter from the origin of the cystic artery was 20 mm (minimum of 2.1 mm and maximum of 53 mm) proximally and 10 mm (minimum 1 mm and maximum 51 mm) distally (ns). Conclusions: Leaking in 99mTc-MAA (99mTc - labelled macroaggregated albumin) was related to the presence of additional arteries. Regardless of cystic artery embolization, it is 2.5 times safer to inject microspheres distal to its origin, compared to proximal injection. Cystic artery origin relative to the right hepatic artery division usually necessitates embolization of the former.

Are type I dural arteriovenous fistulas safe? Single-centre experience of endovascular treatment of dural arteriovenous fistulas.

INTRODUCTION: There are mixed reports on the incidence of intracranial haemorrhage in patients with dural arteriovenous fistulas. We assessed new proposed risk factors (i.a. number of outflows and outflow diameter) of intracerebral haemorrhage due to intracranial dural arteriovenous fistula and presented our personal experience in endovascular treatment of dural arteriovenous fistulas. MATERIAL AND METHODS: The patient database from January 2006 and December 2016 was reviewed, and 25 patients with 28 dural arteriovenous fistulas were identified. RESULTS: 50% of patients presented with intracerebral haemorrhage. Multiple dural fistulas occurred in 12% of patients. Spearman's rank correlation coefficient revealed that there was a strong association between Cognard classification type and time needed to treat (r = 0.59, p < 0.05), as well as the volume of contrast used (r = 0.77, p < 0.05). Infratentorial (r = 0.53, p < 0.05) and right-sided (r = 0.66, p < 0.05) localisation were more challenging to treat. Bleeding was associated with poorer clinical outcome (r = 0.48, p < 0.05). No significant differences were found between the non-haemorrhagic group and the haemorrhagic group regarding the number of outflows (p = 0.459) and largest outflow diameter (p = 0.298). Clinical evaluation at follow-up was as follows: 56% of patients were asymptomatic, 24% had non-significant disability, maintaining independency, 16% had moderate disability, and 8% died - one in the course of intracerebral haemorrhage and one due to other sustained injuries. There were no reported embolisation-related complications. CONCLUSIONS: To conclude, regardless of presentation, both symptomatic and asymptomatic dural arteriovenous fistulas deserve clinical attention, structured evaluation, and follow-up. Type I fistulas were associated with haemorrhage in 1/3 of all cases. Overall our results indicate that the risk of haemorrhage and dire consequences is multifactorial.

Superior ophthalmic vein and ophthalmic artery in immediate evaluation after endovascular treatment of carotid-cavernous fistulas.

PURPOSE: To validate superior ophthalmic vein (SOV) and ophthalmic artery (OA) usefulness in immediate evaluation of new endovascular approaches to treat carotid-cavernous fistulas (CCFs). MATERIAL AND METHODS: A retrospective review of 597 intracerebral malformation embolisations yielded 40 embolisations of CCF in the treatment of 18 patients. Two interventional radiologists performed detailed radiological angiographic assessments. RESULTS: Mean age at initial admission was 58.9 years (SD 18.5 years, range 24-85 years). Patients presented with: chemosis (50%), ocular bruit (50%), exophthalmos (61%), diminished visual acuity (77.8%), headache (16.7%), and intracerebral haemorrhage (5.55%), and 5.55% were asymptomatic. Unilateral fistulas (10-55.5%) showed more diversified venous drainage pattern than bilateral ones (8-44.4%). There were statistically significant differences in post-traumatic and spontaneous CCF regarding age (p = 0.036), type of fistula (p = 0.0008), and presence of pseudoaneurysm (p = 0.036). 77.8% of patients had increased ipsilateral SOV diameter. SOV enlargement was not associated with type of fistula, history of trauma, or degree of exophthalmos. Ipsilateral ophthalmic artery was visible in all patients on both pre- and postprocedural angiography on lateral projection. Pre- and post-procedural SOV diameter was significantly different. Internal carotid artery patency was 100%, while the overall final angiographic or clinical success was 85.7%. We had three cases of peri-procedural complications. CONCLUSIONS: We reported changeable dynamics of SOV and OA after endovascular treatment of CCFs and proved the feasibility of coils and Onyx-18 in the treatment thereof.

Short arginine-rich lipopeptides: From self-assembly to antimicrobial activity.

In this paper, we examine antimicrobial and cytotoxic activities, self-assembly and interactions with anionic and zwitterionic membranes of short arginine-rich lipopeptides: C16-RRRR-NH2, C14-RRRR-NH2, C12-RRRR-NH2, and C16-PRRR-NH2. They show a tendency to self-assembly into micelles, but it is not required for antimicrobial activity. The membrane binding of the lipopeptides can be accompanied by other factors such as: peptide aggregation, pore formation or micellization of phospholipid bilayer. The shortening of the acyl chain results in compounds with a lower haemolytic activity and a slightly improved antimicrobial activity against Gram-positive bacteria, what indicates enhanced cell specificity. Results of coarse-grained molecular dynamics simulations indicate different organization of membrane lipids upon binding of arginine-based lipopeptides and the previously studied lysine-based ones.

The Predictive Value of SPECT/CT imaging in colorectal liver metastases response after 90Y-radioembolization.

CONCLUSION: The mT/N1 ratio, PAD, and AAD can be used as predictors of tumor response to SIRT treatment, and SPECT/CT imaging can be used for dosimetric assessment of radioembolization.

Interventional radiology treatments for iatrogenic severe bleeding during percutaneous coronary interventions.

PURPOSE: Interventional cardiology and interventional radiology are separate medical disciplines in which intra-arterial contrast media are used. Interventional cardiology has resigned from many types of treatment techniques that are now used and developed in the field of interventional radiology. In the event of iatrogenic bleeding during coronary interventions, there is an urgent need to use safe and efficient rescue procedures that are as efficient as cardiosurgery but use simpler treatment options. Serious perforations require immediate endovascular interventions. Medical history may reveal risk factors for artery perforation. Medicines, location of artery perforation, and extent of bleeding are directly associated with the prognosis. Most often, arterial perforations are due to inappropriate wire manipulation or use of oversized balloons or cutting balloons. Prolonged, artery-occluding balloon inflation, covered stent implantation, and embolisation with different agents are among the available treatment options for artery ruptures. MATERIAL AND METHODS: A retrospective analysis was carried out among selected patients with iatrogenic vascular complications during procedures involving either coronary or non-coronary arteries. RESULTS: Only representative cases were selected and presented in the patient subsection. CONCLUSIONS: Artery perforation during cardiac catheterisation can lead to dire consequences. To manage this complication, clinicians need pre-established procedures, adequate resources, and knowledge. Interventional radiology can be used as a salvage therapy in such cases.

[Treatment of ischemic stroke with mechanical trombectomy systems]. / Leczenie udaru niedokrwiennego mózgu z zastosowaniem systemów do trombektomii mechanicznej.

The implementation of ischemic stroke therapy has created new opportunities for clinical improvement and the reversal of adverse prognosis in patients with ischemic stroke. Mechanical thrombectomy has become the recommended treatment for acute stroke in a select group of patients and in highly specialized centres with experience in endovascular therapy. AIM: The aim of the study was to evaluate of the efficacy and safety of mechanical thrombectomy in patients with acute ischemic stroke within the first six hours of illness was reported. MATERIALS AND METHODS: 34 patients previously hospitalized in the Department of Neurology due to ischemic stroke were included in the study during the first six hours of illness. Short-term efficacy and safety (1 month after surgery) and long-term (3 months) were evaluated based on the assessment of early mortality, functional status and neurological status. Factors that increase the risk of death were also analyzed. RESULTS: In the study group, the recanalization of the vessel was obtained in 52% of patients, which was associated with a significant improvement of functional status. Improvements in functional and neurological status were obtained in most of the patients (63%), including very good functional status (mRS 0-1) in 7 (20%) patients upon discharge from the hospital. CONCLUSIONS: In the study group, the recanalization of the vessel was obtained in 52% of patients, which was associated with a significant improvement of functional status. Improvements in functional and neurological status were obtained in most of the patients (63%), including very good functional status (mRS 0-1) in 7 (20%) patients upon discharge from the hospital.

PEGylated substrates of NSP4 protease: A tool to study protease specificity.

Herein we present the synthesis of a novel type of peptidomimetics composed of repeating diaminopropionic acid residues modified with structurally diverse heterobifunctional polyethylene glycol chains (abbreviated as DAPEG). Based on the developed compounds, a library of fluorogenic substrates was synthesized. Further library deconvolution towards human neutrophil serine protease 4 (NSP4) yielded highly sensitive and selective internally quenched peptidomimetic substrates. In silico analysis of the obtained peptidomimetics revealed the presence of an interaction network with distant subsites located on the enzyme surface.

Investigation of Serine-Proteinase-Catalyzed Peptide Splicing in Analogues of Sunflower Trypsin Inhibitor†1 (SFTI-1).

Serine-proteinase-catalyzed peptide splicing was demonstrated in analogues of the trypsin inhibitor SFTI-1: both single peptides and two-peptide chains (C- and N-terminal peptide chains linked by a disulfide bridge). In the second series, peptide splicing with catalytic amount of proteinase was observed only when formation of acyl-enzyme intermediate was preceded by hydrolysis of the substrate Lys-Ser peptide bond. Here we demonstrate that with an equimolar amount of the proteinase, splicing occurs in all the two-peptide-chain analogues. This conclusion was supported by high resolution crystal structures of selected analogues in complex with trypsin. We showed that the process followed a direct transpeptidation mechanism. Thus, the acyl-enzyme intermediate was formed and was immediately used for a new peptide bond formation; products associated with the hydrolysis of the acyl-enzyme were not observed. The peptide splicing was sequence- not structure-specific.

Superselective renal artery embolization in the treatment of iatrogenic bleeding into the urinary tract.

BACKGROUND: The aim of this study was to evaluate the efficacy of superselective renal artery embolization in patients with bleeding into the urinary system. MATERIAL/METHODS: From 2007 to 2012, 20 patients were treated with superselective renal artery embolization for bleeding after percutaneous nephrolithotomy (PCNL), nephron-sparing surgery (NSS), including 1 patient with AVF after PCNL. During the procedure, embolization material was injected through a microcatheter to stop the bleeding. Embolization materials included a mixture of cyanoacrylate and lipiodol, embolization coils, and Spongostan. Clinical evaluation included remission of hematuria and normalization of blood morphotic elements. RESULTS: The cause of bleeding into the urinary tract was damage to vessels (all cases): with coexisting false aneurism (8 cases); with coexisting arterio-venus fistula (1 case); and with coexisting intrarenal hematoma (3 cases). The bleeding occurred 2-5 days after PCNL and NSS, and 10 days after PCNL with AVF. The mean hematocrit level was 22%-24%. Technical success was achieved in 20 cases. Clinical success was achieved in 19 cases. One patient with hematuria after PCNL with AVF needed a second endovascular treatment to stop bleeding. CONCLUSIONS: Superselective renal artery embolization is an effective procedure in the treatment of iatrogenic bleeding into the urinary tract after PCNL and NSS.

Combined thrombolysis in posterior circulation stroke caused by bilateral vertebral artery dissection in squash player.

BACKGROUND AND PURPOSE: Growing number of vertebral artery dissection has been detected due to higher awareness and improved imaging techniques, even after seemingly minor head turning in young- or middle-aged adults without predisposing risk factors for cerebrovascular disease. Treatment options for this condition at this time are limited and there is a troubling shortage of controlled studies. SUMMARY OF CASE: Our patient suffered a bilateral vertebral artery dissection complicated by posterior circulation stroke. We decided to treat acute stroke with intravenous thrombolytic therapy. Patient's condition worsened despite the treatment so emergency angiography was performed to assess the arterial patency. Additional dose of recombinant tissue plasminogen activator together with mechanical thrombectomy was administered using intraarterial route. The patient recovered well and at 12-month follow-up visit he had only right marginal incomplete hemianopia. CONCLUSIONS: Vertebral artery dissection should be taken into consideration in differential diagnosis of posterior circulation stroke or TIA in young patients with a history of even as subtle precipitating events as forceful head movements. Combined thrombolytic therapy may provide safe and effective treatment of stroke-complicated cases. This case report shows that expanded diagnostic protocol for acute ischemic stroke, including computed tomography perfusion study and angiography of cervical and cranial vessels, assures rapid and correct diagnosis.