Chiral NHC Ligands for Enantioselective Gold(I) Catalysis Under Aerobic Conditions: the Importance of Conformational Flexibility and Steric Hindrance of NHC Ligand on Reactivity.

Gold(I) catalysis has been recognized as a valuable tool for the unique transformation of multiple carbon-carbon bonds. Enantioselective π-catalysis based on gold(I) complexes is, however, still underdeveloped due to lack of privileged ligands. Herein, we present an accessible method to a new family of stable yet catalytically active chiral NHC-Au(I)-Cl complexes. The key to preserving a simultaneous fine balance between reactivity and stability in this newly developed family appears to be sterically hindered, but conformationally flexible NHC ligands. These could be easily accessed on a multigram scale by merging sterically hindered anilines with commercially available amino alcohols and amines via a four-steps synthetic sequence without the need for chromatographic purification. Further investigations of the catalytic activity of NHC-Au-Cl complexes identified the OH functionality incorporated into the NHC core as crucial for the level of enantioselectivity as well as the TsO- anion responsible for the activation of NHC-Au(I)-Cl. Finally, NMR studies and X-ray investigations revealed for the first time that the widely accepted ion metathesis (NHC-Au-Cl to NHC-Au-OSO2 R) responsible for the activation of NHC-Au-Cl complexes does not take place (or it is very slow) in commonly used MeNO2 in contrast to DCM.

The effect of shift work on body mass index: A systematic review and meta-analysis of observational studies.

CONTEXT: Shift work involves working outside the standard working hours of 9 am to 5 pm Monday to Friday and may include working evening, night, weekend, or rotating shift patterns. Since shift workers sleep less and their circadian rhythms are disrupted, they are reported to have higher body weight than day workers. OBJECTIVE: This meta-analysis aims to determine whether shift workers (SW) are more prone to higher body mass index (BMI) than their day workers (DW) counterparts. It also addresses the question of whether the duration of shift work exposure, sex, or occupational type affect BMI value. METHODS: Four databases that is PubMed, EBSCO Host, Scopus, and Web of Science were searched for reports published up to October 2023. RESULTS: Sixty-three studies involving a total of 693 449 participants met our inclusion criteria. Meta-analyses showed a significant effect of shift work on BMI value (standard mean difference; SMD: 0.10 kg/m2 [95% confidence interval; 95% CI: 0.07; 0.13; p < .001]) as compared with non-shift counterparts. Subgroup analysis revealed that shift work significantly increased BMI for studies where male working on this job schedule (SMD: 0.10 kg/m2 [95% CI: 0.04; 0.17; p = .0018]) for studies where shift workers worked ≥13 years (calculated as the median of shift work experience; SMD: 0.14 kg/m2 [95% CI: 0.10; 0.18; p < .001]) as well as for studies where industrial (SMD: 0.12 kg/m2 [95% CI: 0.05; 0.19; p = .0012]) and other type of occupations were dominated (0.12 kg/m2 [95% CI: 0.07; 0.16; p < .001]). CONCLUSIONS: We found that in general working on a shift schedule increases BMI as compared with day workers, especially in case of male employed in this job schedule, for shift workers working for long periods of time (≥13 years), and for shift workers employed in industry and other type of occupations(e.g., airline workers, firefighters, police officers, blue collar, security personnel, bus drivers, garbage collectors, railway workers, postal, oil and gas workers).

Inhaler use technique course: an effective postgraduate training solution for pharmacists to enhance therapeutic outcomes as part of patient education.

BACKGROUND: Patients with asthma and chronic obstructive pulmonary disease could benefit from education on using inhalers provided by pharmacists. However, pharmacists may have limited competencies, indicating the necessity to implement appropriate postgraduate courses. The study aimed to evaluate an inhaler use course for pharmacists, including its impact on participants' knowledge and satisfaction. METHODS: The study involved 261 pharmacists from community pharmacies and was conducted between September 2019 and March 2021. A pre-post analysis of their knowledge of the topic was applied. Additionally, at the beginning of the course, participants were asked about their educational needs, and at the end, they completed a satisfaction survey. The preferred learning formats indicated by participants were interactive workshops and lectures. RESULTS: As a result of the course, both their actual and self-assessed level of knowledge significantly increased. The percentage of correct answers in the test before the training was 24.4%, while after, it was 84.3% (p < 0.0001). Before the course, their average self-assessed level of knowledge was 52.0%, and after the training, it increased to 90.0% (p < 0.0001). Almost all respondents stated that the course met their expectations. They estimated their satisfaction at 94.0% and the usefulness of the provided information at 98.0%. CONCLUSIONS: Improved preparation of pharmacists resulting from their participation in the course can contribute to providing more professional advice to patients, thereby positively influencing the pharmaceutical care process in community pharmacies.

Vitamin D supplementation increases objective response rate and prolongs progression-free time in patients with advanced melanoma undergoing anti-PD-1 therapy.

BACKGROUND: Vitamin D3 is a prohormone with pleiotropic effects, including modulating the functions of the immune system and may affect the effectiveness of anti-PD-1 treatment in patients with cancer. According to the literature, the potential mechanism of vitamin D's influence on the effectiveness of therapy is most likely related to the amount and activity of tumor-infiltrating lymphocytes. There are data showing the effect of vitamin D on cells regulating the activity of CD8 lymphocytes. METHODS: A total of 200 patients with advanced melanoma were included in the study. All patients received anti-PD-1 immunotherapy (nivolumab or pembrolizumab) as first-line treatment. Serum vitamin D levels were measured in patients both before and every 12 weeks during treatment. Part of the group had vitamin D measured retrospectively from the preserved serum. The other part of the supplementation group was tested prospectively. RESULTS: The response rate in the group with low vitamin D levels and not supplemented was 36.2%, whereas in the group with normal baseline levels or a normal level obtained with supplementation was 56.0% (p = .01). Moreover, progression-free survival in these groups was 5.75 and 11.25 months, respectively (p = .03). In terms of overall survival, there was also a difference in favor of the group with normal vitamin D levels (27 vs. 31.5 months, respectively; p = .39). CONCLUSIONS: In our opinion, maintaining the vitamin D level within the normal range during anti-PD-1 immunotherapy in advanced melanoma patients should be a standard procedure allowing the improvement of treatment outcomes.

Physical activity, quality of diet and bone mineral density in patients with inflammatory bowel disease.

BACKGROUND: The aim of this study was to find an association between moderate, vigorous and total physical activity (PA); diet quality; and bone mineral density (BMD) among patients suffering from inflammatory bowel disease (IBD). METHODS: We enrolled 54 IBD patients, including those with Crohn's disease (CD) and ulcerative colitis (UC), and 24 healthy adults. All subjects completed the Questionnaire of Eating Behaviour based on which prohealthy and nonhealthy diet indexes were calculated, and the questionnaire included questions from the International Physical Activity Questionnaire. Prohealthy and nonhealthy diet indexes were divided into low-, medium- and high scores. BMD and T- and Z-scores of the lumbar spine (L1-L4) and femoral neck (FN) were assessed using dual-energy X-ray absorptiometry method. RESULTS: BMD, T- and Z-scores of the FN and the Z-score of L1-L4 were significantly lower among patients with CD and UC than healthy controls. We did not find any differences in the time of PA among CD, UC and control groups (CG). The prohealthy diet index was higher among healthy subjects than the CD and UC groups. The nonhealthy diet index was lower among UC patients compared with the CG or CD patients. Prohealthy diet index positively correlated with BMD and T- and Z-scores of L1-L4 and FN in IBD. The prohealthy diet index correlated negatively with C-reactive protein and positively with body mass index. The prohealthy diet index correlated only with total PA in the CD group. CONCLUSION: A well-balanced diet and proper PA may decrease the risk of osteoporosis in IBD, so education of patients referring to nutrition and PA is needed.

Impact of the COVID-19 pandemic and lockdown on mental health among the general population in Poland and on the availability of psychological support.

The objective of this paper was to assess the impact of the COVID-19 pandemic on the mental health of Poles and the availability of psychiatric and psychological support during the pandemic. A prospective, cross-sectional web-based survey design was adopted. It was carried out on a sample of 1080 Polish citizens above 18 years old. The majority of Poles (70%) experienced symptoms of mental disorders during the COVID-19 pandemic. Sleep disturbance (63.79%), anxiety (48.79%), low mood for most of the day (44.63%), sadness, tearfulness (41.29%), and concentration disorders (35.93%) were the mental disorder symptoms reported most often by the survey subjects. To reduce the potential acute and long-lasting effects of the COVID-19 pandemic on mental health, decision-makers of the Polish health system should implement stepped multilevel and multi-target psychological support services.

NHC-BIAN-Cu(I)-Catalyzed Friedländer-Type Annulation of 2-Amino-3-(per)fluoroacetylpyridines with Alkynes on Water.

The direct catalytic alkynylation/dehydrative cyclization of 2-amino-3-trifluoroacetyl-pyridines on water was developed for the efficient synthesis of a broad range of fluorinated 1,8-naphthyridines from terminal alkynes. A novel N-heterocyclic carbene (NHC) ligand system that combines a π-extended acenaphthylene backbone with sterically bulky pentiptycene pendant groups was successfully utilized in a copper- or silver-mediated cyclization. Computational analysis of the reaction pathway supports our explanation of the different experimental conversions and yields for the set of copper and silver catalysts. The impact of steric hindrance at the metal center and the flexibility of substituents on the imidazole ring of the NHC on catalytic performance are also discussed.

Artificial miRNAs targeting CAG repeat expansion in ORFs cause rapid deadenylation and translation inhibition of mutant transcripts.

Polyglutamine (polyQ) diseases are incurable neurological disorders caused by CAG repeat expansion in the open reading frames (ORFs) of specific genes. This type of mutation in the HTT gene is responsible for Huntington's disease (HD). CAG repeat-targeting artificial miRNAs (art-miRNAs) were shown as attractive therapeutic approach for polyQ disorders as they caused allele-selective decrease in the level of mutant proteins. Here, using polyQ disease models, we aimed to demonstrate how miRNA-based gene expression regulation is dependent on target sequence features. We show that the silencing efficiency and selectivity of art-miRNAs is influenced by the localization of the CAG repeat tract within transcript and the specific sequence context. Furthermore, we aimed to reveal the events leading to downregulation of mutant polyQ proteins and found very rapid activation of translational repression and HTT transcript deadenylation. Slicer-activity of AGO2 was dispensable in this process, as determined in AGO2 knockout cells generated with CRISPR-Cas9 technology. We also showed highly allele-selective downregulation of huntingtin in human HD neural progenitors (NPs). Taken together, art-miRNA activity may serve as a model of the cooperative activity and targeting of ORF regions by endogenous miRNAs.

Neutrophil Count as Atrioventricular Block (AVB) Predictor following Pediatric Heart Surgery.

Neutrophils play a significant role in immune and inflammatory reactions. The preoperative inflammatory activation may have a detrimental effect on postoperative outcomes. The aim of the study was to investigate the relation between preoperative hematological indices on postoperative complications' risk in pediatric cardiac congenital surgery. The retrospective single center analysis included 93 pediatric patients (48 (65%) males and 45 (35%) females), mean age of 7 (3−30) months referred for cardiac surgery in cardiopulmonary bypass due to functional single ventricle disease (26 procedures), shunts lesions (40 procedures) and cyanotic disease (27 procedures). Among simple hematological indices, the receiver-operating-characteristic curve showed that a neutrophil count below 2.59 K/uL was found as an optimal cut-off point for predicting postoperative atrioventricular block following pediatric cardiac surgery (AUC = 0.845, p < 0.0001) yielding a sensitivity of 100% and a specificity of 65.62%. Preoperative values of neutrophil count below 2.59 K/uL in whole blood analysis can be regarded as a predictive factor (AUC = 0.845, p < 0.0001) for postoperative atrioventricular block in pediatric cardiac surgery.

Base-Catalyzed, Solvent-Free Synthesis of Rigid V-Shaped Epoxydibenzo[b,f][1,5]diazocines.

A novel method for the synthesis of epoxydibenzo[b,f][1,5]diazocines exhibiting a V-shaped molecular architecture is reported. The unique approach is based on unprecedented base-catalyzed, solvent-free autocondensation and cross-condensation of fluorinated o-aminophenones. The structure of the newly synthesized diazocines was confirmed independently by X-ray analysis and chiroptical methods. The rigidity of the diazocine scaffold allowed for the separation of the racemate into single enantiomers that proved to be thermally stable up to 140 °C. Furthermore, the inertness of the diazocine scaffold was demonstrated by performing a series of typical transformations, including transition metal-catalyzed reactions, proceeding without affecting the bis-hemiaminal subunit.

Impact of patient knowledge on hypertension treatment adherence and efficacy: A single-centre study in Poland.

Introduction: Recent studies show that treatment of arterial hypertension is unsuccessful. This is due to the patients' insufficient knowledge of about the therapeutic methods and the consequences of not treating arterial hypertension. Objectives: The aim of the study was to evaluate the patients' knowledge concerning therapeutic options, prophylaxis, and complications of arterial hypertension. The study also assessed the effect of such knowledge on hypertension treatment adherence and efficacy. Patients and Methods: The survey included 488 patients (250 female and 238 male), aged over 18 years, diagnosed with and treated in outpatient and inpatient settings at selected healthcare institutions in Poland. A custom-made questionnaire, based on references on this subject, was the key tool in the present study. Information about the course of the disease and evaluation of hypertension treatment efficacy was based on the patients' medical records. Results: The study found that 54.7% of the subjects had good knowledge about arterial hypertension, 40.0% had average knowledge, and 5.3% had poor knowledge. The extent of knowledge about the disease was significantly dependent on the level of education and the place of receiving medical care (p< 0.05). Good knowledge was significantly associated with controlled blood pressure, number of antihypertensive drugs used, frequency of hospitalization, as well as with medication adherence, and healthy lifestyle behaviours (p< 0.05). Conclusions: More than half of the patients presented good knowledge but a large group still had poor knowledge, especially patients with a low level of education and with hypertension treated at a general practitioner's clinic. The results of our study clearly show that knowledge about arterial hypertension affects medication adherence and healthy lifestyle behaviours and improves hypertension treatment efficacy.

Vitamin D, Vitamin D Receptor (VDR) Gene Polymorphisms (ApaI and FokI), and Bone Mineral Density in Patients With Inflammatory Bowel Disease.

In the etiology of inflammatory bowel disease (IBD) and osteoporosis, the connecting element is the involvement of environmental and genetic factors. Vitamin D receptor (VDR) gene polymorphisms may be associated with the pathogenesis of IBD and bone mineral density (BMD). The study aimed to analyze the relationship between ApaI and FokI polymorphisms of the VDR gene, serum vitamin D concentration, and BMD in patients with IBD. The studied group consisted of 172 patients (85 with Crohn's disease [CD], 87 with ulcerative colitis [UC], and 39 healthy subjects - control group [CG]) were examined. Lumbar spine densitometry (L1-L4) and the femoral neck (FN) measurements were performed using dual-energy X-ray absorptiometry (DXA). Serum concentrations of 25-hydroxyvitamin D were determined using electrochemiluminescence binding assay (ECLIA). Polymorphisms were determined with polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP). . We found no statistically significant differences in vitamin D concentration between the 3 studied groups. CD patients who were FF homozygotes had significantly lower FN BMD than FF homozygous from CG (p-value < 0.05). CD patients who were Aa heterozygotes had significantly lower lumbar spine (L2-L4) BMD than Aa heterozygotes from CG (p-value < 0.05). Among patients with the same polymorphic variants, but belonging to different studied groups, statistically significant differences in bone mineral density in the lumbar spine and the closer end of the femoral neck were observed. We consider that it is the disease entity, not the polymorphism variant, may have a decisive impact on BMD.

Neutrophile-to-Lymphocyte Ratio as a Predictor of Mortality and Response to Treatment in Invasive Aspergillosis among Heart Transplant Recipients-Exploratory Study.

Background and objective: Aspergillus pulmonary infections are potentially life-threatening complications that can occur after heart transplantation. The aim of the study was to find an easily available mortality predictor during Aspergillosis infection therapy following heart transplantation. Materials and methods: This study involved 15 heart recipients with the mean age of 55 ± 6 years who were diagnosed with invasive aspergillosis (IA) in a mean time of 80 ± 53 (19-209) days after orthotropic heart transplantation. Results: Out of fifteen patients diagnosed with IA, five died. The mean time from diagnosis to death in the deceased group was 28 ± 18 days. They were diagnosed with IA in a mean time of 80 ± 53 (19-209) days after orthotropic heart transplantation. During the initial seven days of therapy, the neutrophil to lymphocyte ratio (NLR) significantly differed between the two groups on day three and day seven, with median values of 10.8 [4.3-17.0] vs. 20.2 [17.4-116.8] (p = 0.0373) and 5.2 [3.2-8.1] vs. 32.2 [13.5-49.9] (p = 0.0101) in the survivor and the deceased group, respectively. The NLR was a significant predictor of death both on day three (cut-off point 17.2) and day seven (cut-off point 12.08) of therapy. Conclusions: Findings in our study indicate that NLR may be of predictive value in the estimation of mortality risk or response to treatment among patients with invasive aspergillosis following heart transplantation.

Cardioprotective Effect of Low Level of LDL Cholesterol on Perioperative Myocardial Injury in Off-Pump Coronary Artery Bypass Grafting.

Background and Objectives: Coronary artery disease is still a major cause of death in developed countries. Low-density lipoprotein cholesterol (LDL-C) lowering with statin therapy is a key strategy in major acute coronary events' prevention. The aim of the study was to establish if there is a cardioprotective effect of pre-operative LDL lowering therapy on perioperative myocaridal injury in patients undergoing off-pump coronary artery bypass grafting (CABG). Moreover, the impact of pre-operative LDL level on long term outcome was analysed. Materials and Methods: The retrospective single center analysis included 662 consecutive patients (431 (65%) males and 231 (35%) female, mean age of 65 ± 8) referred for cardiac surgery due to stable chronic coronary syndrome between 2012-2018. The follow up was 9 years. Results: A statistically significant difference was found in postoperative serum Troponin-I for LDL thresholds of 1.8 mmol/L (p = 0.009), 2.6 mmol/L (p = 0.03) and 3.0 mmol/L (p = 0.001). The results indicate that cardioprotective role of LDL is achieved within LDL concentration rate below 1.8 mmol/L (<70 mg/dL). Five patients died perioperatively, whereas 1-year and 9-year overall mortality rates were 4% (n = 28) and 18.6% (n = 123), respectively. Comparing the survival group with diseased, Mann-Whitney U test showed a statistically significant difference in HDL-C (p = 0.007), Troponin (p = 0.009), Castelli index (p = 0.001) and atherogenic index (p = 0.004). Preoperative levels of total cholesterol, LDL-C and HDL-C did not significantly differ between survivors and diseased. The 9-year mortality risk did not differ significantly between subgroups divided according to LDL-C thresholds of 1.4 mmol/L (55 mg/dL), 1.8 mmol/L (70 mg/dL), 2.6 mmol/L (100 mg/dL) and 3.0 mmol/L (116 mg/dL). Conclusions: Preoperative low level of LDL-C cholesterol (below 1.83 mmol/L, 70 mg/dL) has a cardioprotective effect on perioperative myocardial injury in off-pump coronary artery bypass grafting.

Monocyte-to-Lymphocyte Ratio as a Predictor of Worse Long-Term Survival after Off-Pump Surgical Revascularization-Initial Report.

Background and objective: Coronary artery disease is one of the leading causes of deaths nowadays and the trends in diagnosis and revascularization are still in plateau despite well-known factors. Simple whole blood count parameters may be used to measure inflammatory reactions that are involved in processes of atherosclerosis progression. The aim of our study was to analyse the association between simply available hematologic indices and long-term mortality following off-pump coronary artery bypass grafting (OPCAB). Material and Methods: The study group comprised 129 consecutive patients (16 females and 113 males, mean age 66 ± 6 years) who underwent surgical revascularization with off-pump technique between January 2014 and September 2019. The mean follow-up was 4.7 +/-1.9 years. A receiver operating characteristics curve was applied to estimate demographical and perioperative parameters including MLR for mortality. Results: Cox regression analysis revealed chronic pulmonary obstructive disease (HR = 2.86, 95%CI 1.05-7.78), MLR (HR = 3.81, 95%CI 1.45-10.06) and right coronary artery blood flow (HR = 1.06, 95%CI 1.00-1.10) as significant factors predicting increased mortality risk. In the presented model, the MLR > 1.44 on 1st postoperative day was a significant predictor of late mortality after the OPCAB procedure (HR = 3.82, 95%CI 1.45-10.06). Conclusions: Pronounced inflammatory reaction after off-pump surgery measured by MLR > 1.44 can be regarded as a worse long-term prognostic factor.

Exposure to hepatitis E virus in hemodialysis patients from west-central Poland.

Hepatitis E virus (HEV) causes travel-related but also locally acquired infections in industrialized parts of the world, including European countries. Food and blood transfusions are possible sources of transmission. Infections caused by zoonotic variants of the virus (particularly HEV-3) may progress to chronic liver disease in a nonnegligible proportion of immunocompromised people. The aim of this study was to assess the prevalence of serological markers of HEV infection in 189 patients on renal replacement therapy (RRT, currently on hemodialysis, HD) living in west-central Poland and to determine the factors related to HEV exposure in this group. Testing was carried out using commonly used commercial assays (Wantai Biological Pharmacy Enterprise Co, Beijing, China). Anti-HEV IgG was detected in 94 patients (49.7%); none of the participants had anti-HEV IgM or HEV Ag. Patients on RRT (HD) for less than 6 months were significantly more likely to be anti-HEV IgG-positive than dependent of RRT (HD) for more than half a year (80% vs 47%; P = .014). Exposure to HEV in patients from west-central Poland is frequent, but no clear sources of this infection have been identified. There were no serological features of ongoing liver disease caused by HEV in the study subjects.

Active 5' splice sites regulate the biogenesis efficiency of Arabidopsis microRNAs derived from intron-containing genes.

Arabidopsis, miR402 that is encoded within the first intron of a protein-coding gene At1g77230, is induced by heat stress. Its upregulation correlates with splicing inhibition and intronic proximal polyA site selection. It suggests that miR402 is not processed from an intron, but rather from a shorter transcript after selection of the proximal polyA site within this intron. Recently, introns and active 5' splice sites (5'ss') have been shown to stimulate the accumulation of miRNAs encoded within the first exons of intron-containing MIR genes. In contrast, we have observed the opposite effect of splicing inhibition on intronic miR402 production. Transient expression experiments performed in tobacco leaves revealed a significant accumulation of the intronic mature miR402 when the 5'ss of the miR402-hosting intron was inactivated. In contrast, when the miR402 stem-loop structure was moved into the first exon, mutation of the first-intron 5'ss resulted in a decrease in the miRNA level. Thus, the 5'ss controls the efficiency of miRNA biogenesis. We also show that the SERRATE protein (a key component of the plant microprocessor) colocalizes and interacts with several U1 snRNP auxiliary proteins. We postulate that SERRATE-spliceosome connections have a direct effect on miRNA maturation.

As the twig is bent, so is the tree inclined: a survey of student attitudes toward interprofessional collaboration supported with the curricula analysis.

Pharmaceutical care was legally introduced as an interprofessional service in Poland in 2009. However, a collaboration between physicians and pharmacists remains incidental. Proper education at the undergraduate level is necessary to shape the attitudes of students toward establishing interprofessional relations. The aim of the study was to assess the perception of physician-pharmacist collaboration among final-year medical and pharmacy students through questionnaires with both closed-ended and open-ended questions. The study also includes an analysis of medicine and pharmacy curricula in terms of promoting interprofessional collaboration between the two fields. The statistical analysis of data obtained from 502 respondents revealed significant differences between the perceived areas for such collaboration. Moreover, the division of roles and responsibilities during the pharmacotherapy process between both professions seems to be unclear. Importantly, only 10.14% of the respondents evaluated these professional relations as 'good' or 'very good'. Also, 66.87% of the students emphasized the importance of educational interventions to improve interprofessional collaboration between pharmacists and physicians. Although 70% of medical and 87% of pharmacy students wish to establish such collaboration in the future, only 15% and 35%, respectively feel adequately prepared for the task. Understanding similarities and differences in this field appears to be the key to designing effective educational solutions for promoting interprofessional attitudes among healthcare undergraduates.

Double atrial potentials in left-sided accessory pathways are associated with paroxysmal atrial fibrillation.

INTRODUCTION: Muscular connections between the coronary sinus (CS) and left atrium probably impact distribution of electrical activity. Double atrial potentials (DP) may be their presentation. The aim was to investigate the presence of DP in CS recordings during atrioventricular reentrant tachycardia (AVRT) and its contribution to the occurrence of paroxysmal atrial fibrillation (AF). METHODS: A group of 247 patients with accessory pathways (AP) were screened for DP. The patients with DP during AVRT were compared to those without DP. RESULTS: DP during AVRT were found only among the left-sided AP (AP-L). Patients with AP-L were divided into Group 1 (n  =  17) with DP during AVRT and Group 2 (n  =  108) without DP. Patients in Group 1 had higher incidence of AF in history (47.1% vs. 23.1%; P  =  0.0376), AF induced during electrophysiological (EP) study (70.6% vs. 25%; P  =  0.0002). Group 1 had higher heart rate (HR) during AVRT in the EP study (197.2 ± 27 vs. 175.1 ± 26.3 bpm; P  =  0.0019), but HR of clinical AVRT (208.5 ± 30.8 vs. 191.6 ± 27.8 bpm) was not significant different (P  =  ns). Additionally, electrical alternans of QRS amplitude during AVRT in the EP study was more frequent in Group 1 (52.9 vs. 20.4 %; P  =  0.0048). CONCLUSION: Patients with DP and AP-L were more prone to develop AF. The presence of DP was associated with faster AVRT rate. The direction of atrium depolarization during AVRT may be different in the presence of DP and probably plays a role in development of AF in this group of patients.

2D and 3D FISH of expanded repeat RNAs in human lymphoblasts.

The first methods for visualizing RNAs within cells were designed for simple imaging of specific transcripts in cells or tissues and since then significant technical advances have been made in this field. Today, high-resolution images can be obtained, enabling visualization of single transcript molecules, quantitative analyses of images, and precise localization of RNAs within cells as well as co-localization of transcripts with specific proteins or other molecules. In addition, tracking of RNA dynamics within single cell has become possible. RNA imaging techniques have been utilized for investigating the role of mutant RNAs in a number of human disorders caused by simple microsatellite expansions. These diseases include myotonic dystrophy type 1 and 2, amyotrophic lateral sclerosis/frontotemporal dementia, fragile X-associated tremor/ataxia syndrome, and Huntington's disease. Mutant RNAs with expanded repeats tend to aggregate predominantly within cell nuclei, forming structures called RNA foci. In this study, we demonstrate methods for fluorescent visualization of RNAs in both fixed and living cells using the example of RNAs containing various expanded repeat tracts (CUG, CCUG, GGGGCC, CGG, and CAG) from experiment design to image analysis. We describe in detail 2D and 3D fluorescence in situ hybridization (FISH) protocols for imaging expanded repeats RNAs, and we review briefly live imaging techniques used to characterize RNA foci formed by mutant RNAs. These methods could be used to image the entire cellular pathway of RNAs, from transcription to degradation.