Fabrication of electro-microfluidic channel for single cell electroporation

The point of this paper is to demonstrate the use of a quick and cheap fabrication method to realize a laser-ablated microfluidic channel for single cell electroporation. Traditional lithography of microchannel with electrode in MEMS applications has always been complicated. Here, we introduce a new methodology of fabricating microchannel with electrical functionalities achieved through a fast and cheap process. In the present methodology, the microchannel pattern is cut out of polyimide, bonded to two ITO-coated substrates using Teflon as an adhesion layer. ITO as conductive material enables electric field in the channel and its optical transparency allows microscopy techniques to be utilized in characterizing the behavior of the microfluidic chip. The performance of the chip was tested on irreversible single-cell scale electroporation which requires relatively high voltages. CHO cells, as mammalian cells, were passed through the microchannel to experience electric field. Cells were loaded with a fluorogenic dye, Calcein AM, and the electroporation of each was individually recorded in real-time via fluorescent microscopy. The results show promising performance of the electric microchannel in electroporation. By customizing of ITO electrodes and the design of microchannel pattern, utilization and integration of the proposed electrical microchannel in variety of other MEMS-based devices are achievable.     

Child and adolescent psychiatry training and mental health care in Southeast Europe

European Child & Adolescent Psychiatry - There is very limited information available on child and adolescent psychiatry (CAP) training in the Southeast European (SEE) region. The objective of...     

Predictors of Increased Left Ventricular Filling Pressure in Dialysis Patients with Preserved Left Ventricular Ejection Fraction

Aim

To study the left and right ventricular function and to assess the predictors of increased left ventricular (LV) filling pressure in dialysis patients with preserved LV ejection fraction.

Methods

This study included 63 consecutive patients (age 57 ± 14 years, 57% women) with end-stage renal failure. Echocardiography, including tissue Doppler measurements, was performed in all patients. Based on the median value of the ratio of transmitral early diastolic velocity to early myocardial velocity (E/E’ ratio), patients were divided into 2 groups: the group with high filling pressure (E/E’>10.16) and the group with low filling pressure (E/E’≤10.16).

Results

Compared with patients with low filling pressure, the group of patients with high filling pressure included a higher proportion of diabetic patients (41% vs 13%, P = 0.022) and had greater LV mass index (211 ± 77 vs 172 ± 71 g/m³, P = 0.04), lower LV lateral long axis amplitude (1.4 ± 0.3 vs 1.6 ± 0.3 cm, P = 0.01), higher E wave (84 ± 19 vs 64 ± 18cm/s, P < 0.001), lower systolic myocardial velocity (S’:8.6 ± 1.5 vs 7.0 ± 1.3 cm/s, P < 0.001), and lower diastolic myocardial velocities (E’: 6.3 ± 1.9 vs 9.5 ± 2.9 cm/s, P < 0.001; A’: 8.4 ± 1.9 vs 9.7 ± 2.5 cm/s, P = 0.018). Multivariate analysis identified LV systolic myocardial velocity – S’ wave (adjusted odds ratio, 1.909; 95% confidence interval, 1.060-3.439; P = 0.031) and age (1.053; 1.001-1.108; P = 0.048) as the only independent predictors of high LV filling pressure in dialysis patients.

Conclusions

In dialysis patients with preserved left ventricular ejection fraction, reduced systolic myocardial velocity and elderly age are independent predictors of increased left ventricular filling pressure.Cardiovascular disorders are the main cause of mortality and morbidity in patients with end-stage renal failure who are in regular hemodialysis programs (1,2). The left ventricular (LV) hypertrophy is a common finding in these patients. It reflects a physiological response to pressure and volume overload (3) and positively correlates with cardiovascular mortality (4). LV hypertrophy is frequently associated with LV dilatation and reduced systolic function (5). An increased incidence of atherosclerotic cardiovascular events in these patients has also been reported (6). Systolic dysfunction and LV hypertrophy have been identified as the best predictors of outcome in dialysis patients (4,7,8). However, the conventional systolic dysfunction appears in the late stages of the chronic renal failure (9).In contrast to conventional echocardiography, tissue Doppler imaging of the myocardial velocities overcomes the load dependence of diastolic parameters (10). The ratio of transmitral early diastolic velocity (E) to early myocardial velocity (E’) (E/E’ ratio) has been shown to be an accurate method of the LV filling pressure estimation (8) and the best predictor of LV diastolic filling in various cardiac pathologies (11,12), thereby serving as one of the best predictors of outcome in heart failure patients (13-15) and patients with end-stage renal disease (16).The aims of this study were to investigate the left and right ventricular function in patients with end-stage renal disease and preserved LV ejection fraction and to assess the predictors of increased LV filling pressure in these patients.     

Phytochemical composition,anti-inflammatory activity and cytotoxic effects of essential oils from three Pinus spp.

Context: Inflammation and cell differentiation lead to a number of severe diseases. In the recent years, various studies focused on the anti-inflammatory and anticancer activity of essential oils (EOs) of numerous plants, including different Pinus species.

Objective: The phytochemical composition, anti-inflammatory and cytotoxic activity of EOs from needles and twigs of Pinus heldreichii Christ (Pinaceae) and P. peuce Griseb., and from needles, twigs and cones of P. mugo Turra were determined.

Materials and methods: For separation and identification of the EOs, gas chromatography/flame ion detector (GC/FID) and GC/mass spectrometry were performed. The amount of secreted IL-6 in a lipopolysaccharide (LPS)-stimulated macrophage model was quantified (concentration of oils: 0.0001–0.2%, 3?h incubation). Cytotoxicity on the cancer cell lines HeLa, CaCo-2 and MCF-7 were determined using a MTT (Thiazolyl Blue Tetrazolium Bromide) assay (concentration of oils: 0.001–0.1%, 24?h incubation).

Results: The most prominent members in the oils include: δ-3-carene, α-pinene and linalool-acetate (P. mugo); α-pinene, β-phellandrene and β-pinene (P. peuce); limonene, α-pinene and (E)-caryophyllene (P. heldreichii). EOs showed significant cytotoxic effects on cancer cell lines (IC₅₀ 0.007 to >0.1%), with a reduction in cell viability with up to 90% at a concentration of 0.1%, and anti-inflammatory activity (IC₅₀ 0.0008–0.02%) with a reduction of IL-6 secretion with up to 60% at a concentration of 0.01%.

Discussion and conclusion: The EOs of needles and twigs from P. peuce and P. heldreichii as well as of needles, twigs and cones of P. mugo can be considered as promising agents for anticancer and anti-inflammatory drugs.     

Atezolizumab plus bevacizumab versus sorafenib or atezolizumab alone for unresectable hepatocellular carcinoma: A systematic review

BACKGROUNDDespite the use of current standard therapy, the prognosis of patients with unresectable hepatocellular carcinoma (HCC) is poor, with median survival times of 40 mo for intermediate HCC (Barcelona Clinic Liver Cancer [BCLC] stage B) and 6–8 mo for advanced HCC (BCLC stage C). Although patients with early-stage HCC are usually suitable for therapies with curative intention, up to 70% of patients experience relapse within 5 years. In the past decade, the United States Food and Drug Administration has approved different immunogenic treatment options for advanced HCC, the most common type of liver cancer among adults. Nevertheless, no treatment is useful in the adjuvant setting. Since 2007, the multi-kinase inhibitor sorafenib has been used as a first-line targeted drug to address the increased mortality and incidence rates of HCC. However, in 2020, the IMbrave150 trial demonstrated that combination therapy of atezolizumab (anti-programmed death-ligand 1 [PD-L1]) and bevacizumab (anti-vascular endothelial growth factor [VEGF]) is superior to sorafenib, a single anti-programmed death 1/PD-L1 antibody inhibitor used as an anti-cancer monotherapy for HCC treatment.AIMTo conduct a systematic literature review to evaluate the evidence supporting the efficacy and safety of atezolizumab/bevacizumab as preferred first-line drug therapy over the conventional sorafenib or atezolizumab monotherapies, which are used to improve survival outcomes and reduce disease progression in patients with unresectable HCC and non-decompensated liver disease. METHODSA comprehensive literature review was conducted using the PubMed, Scopus, ScienceDirect, clinicaltrials.gov, PubMed Central, Embase, EuropePMC, and CINAHL databases to identify studies that met the inclusion criteria using relevant MeSH terms. This systematic review was conducted according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines and risk of bias (RoB) were assessed using the Cochrane RoB 2 tool and Sevis.RESULTSIn the atezolizumab/bevacizumab group, an improvement in overall tumor response, reduction of disease progression, and longer progression-free survival were observed compared to monotherapy with either sorafenib or atezolizumab. Hypertension and proteinuria were the most common adverse events, and the rates of adverse events were comparable to those with the monotherapy. Of the studies, there were two completed trials and two ongoing trials analyzed using high quality and low bias. A more thorough analysis was only performed on the completed trials.CONCLUSIONTreatment of HCC with atezolizumab/bevacizumab combination therapy was confirmed to be an effective first-line treatment to improve survival in patients with unresectable HCC and non-decompensated liver disease compared to monotherapy with either sorafenib or atezolizumab.     

Epidemiology of hepatitis B virus infection in Albania

AIM： To assess the prevalence and socio-demographic distribution of hepatitis B virus （HBV） infection in Albania. METHODS： Blood samples from 410 unselected schoolboys, 666 students, 500 military personnel, 1286 casual blood donors, 378 voluntary blood donors and 640 pregnant women （total 3880 non-vaccinated residents of rural and metropolitan areas from all over Albania; 2354 （60.7%） male and 1526 （39.3%） female; mean age of 26.3 years） were tested during 2004-2006 for hepatitis B surface antigen （HBsAg） and antibodies to hepatitis B virus （anti-HBs） by ELISA. RESULTS： The HBsAg and anti-HBs prevalence were 9.5% and 28.7%, respectively. The highest HI3sAg prevalence was evident in the younger age group, such as in schoolchildren （11.8%） and the military （10.6%）. Consequently, the anti-HBs prevalence increased with age, from 21.2% in schoolchildren （mean age： 15.7 years）, to 36.3% in pregnant women （mean age： 26.3 years） and 29.7% in voluntary blood donors （mean age： 40.1 years）. There were no significant differences between males and females. CONCLUSION： Despite the estimated two-fold reduction of HBsAg prevalence in the general population from about 18%-19% to 9.5%, Albania remains a highly endemic country （i.e. over 8% of HBsAg prevalence rate）.     

Left atrial systolic antegrade flow in patients with mitral regurgitation: a pulsed Doppler sign displayed by color flow imaging

Twenty-two patients with severe mitral regurgitation were observed to have turbulent systolic antegrade flow on pulsed Doppler mapping of the left atrium. All were studied by color flow imaging to delineate the mechanism of this peculiar flow. Pulsed Doppler findings of an eccentric regurgitant flow in one side, an antegrade systolic flow with slightly delayed onset in the other side, and a low velocity flow near the posterior wall, were consistant with the theory of a large eccentric regurgitant jet swirling in the left atrium. Color flow imaging confirmed this mechanism in all patients. Nineteen patients had flail mitral valve with a positive predictive value of 86%. The other three patients had deformed rheumatic mitral valve. The severity of mitral regurgitation was confirmed in all 16 patients studied by left ventricular cineangiography. We have shown that the antegrade systolic left atrial flow is the result of the swirling of a large regurgitant eccentric jet, is commonly observed with flail mitral valve, can occur in patients with deformed rheumatic mitral valve without flail leaflet, and most significantly indicates the presence of severe mitral regurgitation.     

Factors affecting the quality of bowel preparation for colonoscopy in hard-to-prepare patients: Evidence from the literature

Adequate bowel cleansing is critical for a high-quality colonoscopy because it affects diagnostic accuracy and adenoma detection. Nevertheless, almost a quarter of procedures are still carried out with suboptimal preparation, resulting in longer procedure times, higher risk of complications, and higher likelihood of missing lesions. Current guidelines recommend high-volume or low-volume polyethylene glycol(PEG)/non-PEG-based split-dose regimens. In patients who have had insufficient bowel cleans...