20 years of treating ischemic cardiomyopathy with mesenchymal stromal cells: a meta-analysis and systematic review.

This meta-analysis and systematic review compiles comparative data from 2004 to 2024, investigating the safety and efficacy of mesenchymal stem/stromal cells (MSCs) derived from various tissues for the treatment of ischemic cardiomyopathy (ICM) and associated heart failure. In addition, this review highlights the limitations of these interventions and provides valuable insights for future therapeutic approaches. Relevant articles were retrieved from the PubMed® database using targeted keywords. Our inclusion criteria included clinical trials with patients over 18 years of age, case reports and pilot studies. Animal experiments, in vitro studies, correlational and longitudinal studies, and study designs and protocols were excluded. Forty-nine original articles resulted in follow-up reports of 45 trials. MSCs from bone marrow, umbilical cord and adipose tissue were moderately well tolerated. Of the 1408 participants who received MSCs, 33 trials (67.3%) reported the occurrence of death or serious adverse events. These events resulted in 80 deaths (52% of reported cases) following MSC administration. Importantly, 41.3% of these deaths (n = 33) were not considered to be related to the intervention itself, while 40% of these deaths had no reported cause. As the primary outcome, the mean increase in left ventricular ejection fraction (LVEF) from baseline was 5.75% (95% CI: 3.38% -8.11%, p < 0.0001, I2 = 90,9%) in the randomized controlled trials only (n = 24) within the treatment groups and 3.19% (95% CI: 1.63% to 4.75%, p < 0.0001, I2 = 74,17%) in the control groups after the intervention. When the above results were compared using the standardized mean difference (SDM), a significance in favor of the treatment group was also found (SDM = 0.41; 95% CI: 0.19-0.64, p < 0.001, I2 = 71%). Although improvements were also seen in the control groups, 33.3% (n = 15) of the studies showed no significant difference between the control and treatment groups. The 6-minute walking test (6MWT) and New York Heart Association (NYHA) class scores, used for assessing exercise tolerance and quality of life (QoL), respectively, further supported the improvements in the treatment group. These improvements were noted as 62.5% (n = 10) for the 6MWT and 54.5% (n = 12) for the NYHA class scores. According to the risk of bias analysis, 4 trials were of good quality (11.8%), 15 were of fair quality (44.1%), and 15 were of poor quality (44.1%). Major limitations of these studies included small sample size, diagnostic challenges/lack, uncertain cell dosage and potential bias in patient selection. Despite the ongoing debate surrounding cell administration for ICM, there are supporting signs of improved clinical and laboratory outcomes, as well as improved QoL in the MSC-treated groups. However, it is important to recognize the limitations of each study, highlighting the need for larger, controlled trials to validate these findings.

Vaccine hesitancy among healthcare professionals and the general population: Second important step in the fight against COVID-19.

INTRODUCTION: This study aims at finding valuable information for predicting vaccination intentions against COVID-19 to guide future interventions to address hesitation. METHODOLOGY: This observational study consists of 1010 volunteer health workers from the state hospitals in Bursa, and 1111 volunteers from the non-healthcare group, unvaccinated against COVID-19. In the study, the participants were asked about their sociodemographic information and reasons for refusing the COVID-19 vaccine by face-to-face interview. RESULTS: We classified the unvaccinated healthcare worker group as group 1, and the unvaccinated non-health workers group as group 2. Between groups 1 and 2, vaccination refusal, education level, income level, and pregnancy status were statistically significant (p < 0.001). The groups differed in the reasons for vaccine refusal and recommending vaccination to the relatives of those who refused vaccination (p < 0.001). CONCLUSIONS: Healthcare workers have priority among high-risk groups considered candidates for early vaccination. Therefore, it is important to consider health professionals' attitudes towards COVID-19 vaccination to better address barriers to widespread vaccination. The role of healthcare professionals is also important, as it encourages the entire community to be vaccinated with role-modeling behavior and advises patients and communities.

Can ferritin/lymphocyte percentage ratio, a new indicator, predict the clinical course of COVID-19 cases?

OBJECTIVES: COVID-19 maintains its seriousness as a global emergency with its rapid distribution worldwide. Ferritin / lymphocyte percentage ratio (FLPR) may appear as a prognostic value at the initial evaluation stage and thus can be used as a simple, effective, and reliable parameter in critical patient identification with COVID-19. METHODS: In this retrospective cohort study, we evaluated patients over 18 years old, who were hospitalized after being evaluated as COVID-19 and whose PCR results were positive. We calculated FLPRs from complete blood counts taken during emergency department admissions and classified disease severity due to emergency initial evaluation. The relationship between the severity of the thoracic tomography findings, hospitalization, and intensive care needs, and 28-day mortality with the FLPR were evaluated. RESULTS: The difference between the groups classified according to COVID-19 severity and the FLPR means was statistically significant (x2=148.284; SD=3; p=0.000). FLPR levels were found to be high in critical and serious groups. In the ROC analysis for the FLPR level, the area under the curve (AUC) value was found to be 0.909 (95% CI 0.857-0.961). When the cut off value of FLPR was 9.80, the sensitivity was found to be 97.6 %, and the specificity was 65.2 %, whereas, when the cut off value for FLPR was found to be 21.11, the sensitivity was 82.9 % and the specificity was 82.8 %. CONCLUSION: The FLPR, a new parameter, can be used as a significant marker to predict the 28-day mortality in patients (Tab. 5, Fig. 1, Ref. 25).

The role of emergency department triage early warning score (TREWS) and modified early warning score (MEWS) to predict in-hospital mortality in COVID-19 patients.

BACKGROUND: It is necessary to identify critical patients requiring hospitalization early due to the rapid increase in the number of COVID-19 cases. AIM: This study aims to evaluate the effectiveness of scoring systems such as emergency department triage early warning score (TREWS) and modified early warning score (MEWS) in predicting mortality in COVID-19 patients. METHODS: In this retrospective cohort study, PCR positive patients evaluated for COVID-19 and decided to be hospitalized were evaluated. During the first evaluation, MEWS and TREWS scores of the patients were calculated. Intensive care needs as well as 24-h and 28-day mortality rates were evaluated. RESULTS: A total of 339 patients were included in the study. While 30 (8.8%) patients were hospitalized in the intensive care unit, 4 (1.2%) died in the emergency. The number of patients who died within 28 days was found to be 57 (16.8%). In 24-h mortality, the median MEWS value was found to be 7 (IQR 25-75) while the TREWS value was 11.5 (IQR 25-75). In the ROC analysis made for the diagnostic value of 28-day mortality of MEWS and TREWS scores, the area under the curve (AUC) for the MEWS score was found to be 0.833 (95% CI 0.777-0.888, p < 0.001) while it was identified as 0.823 (95% CI 0.764-0.882, p < 0.001) for the TREWS. CONCLUSION: MEWS and TREWS calculated at emergency services are effective in predicting 28-day mortality in patients requiring hospitalization due to COVID-19.

Predictive Value of Modified Early Warning Scoring System for Identifying Critical Patients with Malignancy in Emergency Department.

BACKGROUND: Identification of critically ill patient is particularly important in the emergency department (ED). The prolonged duration from hospital admission to delivering intensive care service is related to increased mortality. The aim of this study is to evaluate the effectiveness of Modified Early Warning Score (MEWS) for identifying critical patients with malignancy in ED settings. METHODS: We evaluated patients with malignancy who were admitted to our ED of a tertiary university hospital in Turkey over a three-month period. We evaluated MEWS on admission as MEWS 1. After the initial treatment depending on the patients' health status in ED, at 2 hours after admission, we evaluated MEWS again and recorded as MEWS 2. All patients were followed up for 30 days after the initial admission. RESULTS: Mean age (SD) was 59.2 (13.5) and male/female ratio was 295/206. MEWS1 was higher than MEWS2, (MEWS1: 3.05 ± 3.31, MEWS2: 2.35 ± 3.17, P < 0.001). A total of 362 patients (72.3%) survived and 139 (27.7%) died within 30 days of initial admission. MEWS1/MEWS2 values for alive and dead patients were 1.66/0.87, and 6.67/6.21, respectively, and the difference was significant (P < 0.001). ROC analysis was performed for MEWS 1; the area under curve (AUC) for hospitalization was 0.768 (95% CI 0.729 to 0.804) and for mortality was 0.900 (95% CI 0.870 to 0.924). ROC analysis revealed a cut-off value of 2 for predicting both hospitalization and mortality in these patients. The sensitivity of the presented cut-off was 77.32% (72.1%-82.0%) for hospitalization and 76.24% (95% CI 71.5-80.5) for mortality; the specificity was 69.52 (95% CI 62.8-75.7) for hospitalization and 90.65 (95% CI 84.65-94.9) for mortality. CONCLUSION: We found in our study that MEWS evaluation for patients with malignancy on admission to ED is predictive of mortality in the subsequent 30 days, and it is a valuable tool for identifying the critical group. Also, AVPU scores alone can predict mortality in patients admitted to ED.

Hyperbaric oxygen therapy decreases QTc dispersion that increased in CO poisoning.

Myocardial injury is a frequent consequence of moderate to severe CO (carbon monoxide) poisoning and a significant predictor of mortality in CO injury. Electrocardiography (ECG) is an easily accessible diagnostic tool for evaluating myocardial damage. Increased QT interval and QT dispersion are related to heterogeneity of regional ventricular repolarization and can develop into arrhythmias. It has been reported that QT interval and QT dispersion increase in patients with CO poisoning. Hyperbaric oxygen (HBO2) therapy has been used successfully in treating patients with CO poisoning. The aim of this study was to investigate change of corrected QT (QTc) interval and QTc dispersion after HBO2 therapy. This study included 31 patients with CO poisoning. QTc dispersion increased in patients with CO poisoning. The mean QTc dispersion was 54.94 milliseconds (ms) on admission. The mean QTc dispersion decreased to 35.74 ms after HBO2 therapy (P=0.003). There was also a correlation between carboxyhemoglobin level and QTc dispersion (P=0.029). HBO2 therapy, which decreases QTc dispersion, may improve the myocardial electrical homogeneity and reduce the risk of ventricular arrhythmia and cardiac death. Physicians should be aware of the effect of HBO2 therapy on myocardial damage when treating patients with CO poisoning. The ECGs should be examined carefully before referring or excluding HBO2 therapy.

Oligonucleotides suppress PKB/Akt and act as superinductors of apoptosis in human keratinocytes.

DNA oligonucleotides (ODN) applied to an organism are known to modulate the innate and adaptive immune system. Previous studies showed that a CpG-containing ODN (CpG-1-PTO) and interestingly, also a non-CpG-containing ODN (nCpG-5-PTO) suppress inflammatory markers in skin. In the present study it was investigated whether these molecules also influence cell apoptosis. Here we show that CpG-1-PTO, nCpG-5-PTO, and also natural DNA suppress the phosphorylation of PKB/Akt in a cell-type-specific manner. Interestingly, only epithelial cells of the skin (normal human keratinocytes, HaCaT and A-431) show a suppression of PKB/Akt. This suppressive effect depends from ODN lengths, sequence and backbone. Moreover, it was found that TGF alpha-induced levels of PKB/Akt and EGFR were suppressed by the ODN tested. We hypothesize that this suppression might facilitate programmed cell death. By testing this hypothesis we found an increase of apoptosis markers (caspase 3/7, 8, 9, cytosolic cytochrome c, histone associated DNA fragments, apoptotic bodies) when cells were treated with ODN in combination with low doses of staurosporin, a well-known pro-apoptotic stimulus. In summary the present data demonstrate DNA as a modulator of apoptosis which specifically targets skin epithelial cells.

Sequencing of single and double stranded RNA oligonucleotides by acid hydrolysis and MALDI mass spectrometry.

Treatment of RNA oligonucleotides with strong acids at pH 1-2 rapidly leads to hydrolysis of the phosphodiester bonds at the 5'-position of ribose. Analysis of the resulting degradation products by MALDI coupled to an Orbitrap high resolution mass spectrometer shows almost complete mass ladders from both sides of the nucleotides without interfering fragments from base losses or internal fragments. From the mass differences between adjacent peaks of a mass ladder, the sequence can be determined. Low cleavage efficiency at the termini leads to 2mers and 3mers which can be identified by MS/MS. In this way the complete sequences of different siRNA 21mer single and double strands could be verified. This simple and fast method can be applied for controlling sequences of synthetic oligomers, as well as for de-novo sequencing. Moreover, the method is applicable for localization and identification of RNA modifications as demonstrated using the examples of an oligonucleotide with phosphorothioate backbone and of one containing 2'-methoxy-ribose modifications.

Detection and relative quantification of siRNA double strands by MALDI mass spectrometry.

While MALDI-MS is widely accepted for quality control of synthetic oligonucleotides, this method has been regarded as not applicable for a control of the purity and correct annealing of double strands. The results presented here show that the double-strand intensities measured by MALDI-MS maintain and reflect the solution conditions. Using a single-stranded RNA as internal standard, the double-strand intensity can be determined by measuring the intensity ratio of the single strands to the standard under "native" conditions and after denaturation with formic acid. For siRNAs with fully matched 20-21 base pairs, relative intensities of the double strands are between 94 and 97.2%. The stability determined by MALDI-MS for different RNA duplexes correlates well with calculated T m values and the content of G-C pairs. Furthermore, the quantification method enables one to determine an excess of one single strand and the contribution of duplex formation by truncated strands. The results show that MALDI-MS is a fast and reliable method for quality control of synthetic siRNA.

Effect of proline to alanine mutation on the thermal stability of the all-beta-sheet protein tendamistat.

The temperature dependent denaturation of wild-type tendamistat and of the proline-free triple mutant P7A/P9A/P50A was investigated using Fourier-transform infrared (FTIR) spectroscopy. Whereas the temperature-induced unfolding is reversible in the wild type, aggregation was observed for the proline-free tendamistat when studied under the same conditions. The midpoint unfolding temperature T(m) was found as 82.3+/-0.5 degrees C in (2)H2O. The thermal stability of the proline-free mutant is reduced by 15 degrees C as compared to the wild type. Changes in the strength of hydrogen bonding of tyrosine O-H groups upon unfolding and aggregation are reflected in small shifts of the C-C stretching mode of the aromatic ring near 1515 cm(-1). Evaluation of data from different infrared (IR) bands sensitive to changes in secondary structure as well as to changes in tertiary structure strongly supports a two-state model for the unfolding process of wild-type tendamistat.

Structural variations and stabilising modifications of synthetic siRNAs in mammalian cells.

Double-stranded short interfering RNAs (siRNA) induce post-transcriptional silencing in a variety of biological systems. In the present study we have investigated the structural requirements of chemically synthesised siRNAs to mediate efficient gene silencing in mammalian cells. In contrast to studies with Drosophila extracts, we found that synthetic, double-stranded siRNAs without specific nucleotide overhangs are highly efficient in gene silencing. Blocking of the 5'-hydroxyl terminus of the antisense strand leads to a dramatic loss of RNA interference activity, whereas blocking of the 3' terminus or blocking of the termini of the sense strand had no negative effect. We further demonstrate that synthetic siRNA molecules with internal 2'-O-methyl modification, but not molecules with terminal modifications, are protected against serum-derived nucleases. Finally, we analysed different sets of siRNA molecules with various 2'-O-methyl modifications for stability and activity. We demonstrate that 2'-O-methyl modifications at specific positions in the molecule improve stability of siRNAs in serum and are tolerated without significant loss of RNA interference activity. These second generation siRNAs will be better suited for potential therapeutic application of synthetic siRNAs in vivo.

Functional studies of the PI(3)-kinase signalling pathway employing synthetic and expressed siRNA.

RNA interference (RNAi) is a RNA-mediated sequence-specific gene silencing mechanism. Recently, this mechanism has been used to down-regulate protein expression in mammalian cells by applying synthetic- or vector-generated small interfering RNAs (siRNAs). However, for the evaluation of this new knockdown technology, it is crucial to demonstrate biological consequences beyond protein level reduction. Here, we demonstrate that this new siRNA-based technology is suitable to analyse protein functions using the phosphatidylinositol (PI) 3-kinase signal transduction pathway as a model system. We demonstrate stable and transient siRNA-mediated knockdown of one of the PI 3-kinase catalytic subunits, p110beta, which leads to inhibition of invasive cell growth in vitro as well as in a tumour model system. Importantly, this result is consistent with loss-of-function phenotypes induced by conventional RNase H-dependent antisense molecules or treatment with the PI 3-kinase inhibitor LY294002. RNAi knockdown of the downstream kinases Akt1 and Akt2 does not reduce cell growth on extracellular matrix. Our data show that synthetic siRNAs, as well as vector-based expression of siRNAs, are a powerful new tool to interfere with signal transduction processes for the elucidation of gene function in mammalian cells.

Kinetic mechanism and catalysis of a native-state prolyl isomerization reaction.

Folding of tendamistat is a rapid two-state process for the majority of the unfolded molecules. In fluorescence-monitored refolding kinetics about 8% of the unfolded molecules fold slowly (lambda=0.083s(-1)), limited by peptidyl-prolyl cis-trans isomerization. This is significantly less than expected from the presence of three trans prolyl-peptide bonds in the native state. In interrupted refolding experiments we detected an additional very slow folding reaction (lambda=0.008s(-1) at pH 2) with an amplitude of about 12%. This reaction is caused by the interconversion of a highly structured intermediate to native tendamistat. The intermediate has essentially native spectroscopic properties and about 2% of it remain populated in equilibrium after folding is complete. Catalysis by human cyclophilin 18 identifies this very slow reaction as a prolyl isomerization reaction. This shows that prolyl-isomerases are able to efficiently catalyze native state isomerization reactions, which allows them to influence biologically important regulatory conformational transitions. Folding kinetics of the proline variants P7A, P9A, P50A and P7A/P9A show that the very slow reaction is due to isomerization of the Glu6-Pro7 and Ala8-Pro9 peptide bonds, which are located in a region that makes strong backbone and side-chain interactions to both beta-sheets. In the P50A variant the very slow isomerization reaction is still present but native state heterogeneity is not observed any more, indicating a long-range destabilizing effect on the alternative native state relative to N. These results enable us to include all prolyl and non-prolyl peptide bond isomerization reactions in the folding mechanism of tendamistat and to characterize the kinetic mechanism and the energetics of a native-state prolyl isomerization reaction.