Novel insights on GTPBP3-associated hypertrophic cardiomyopathy.

About 100 genes have been associated with cardiomyopathies with genotype-phenotype correlations often hard to establish. Genetic testing may help to confirm the genetic diagnosis and assess the risk of inheritance in the family. A 25-year old male with hypertrophic cardiomyopathy and fasciculoventricular accessory pathway was referred for genetic testing by his cardiologist. Targeted PRKAG2 screening and whole-exome sequencing were performed, followed by Sanger sequencing segregation analysis in the family. The PRKAG2 gene screening was negative. Whole-exome sequencing revealed the following four variants in the patient: c.181G>C (p.Ala61Pro) and c.1199C>T (p.Thr400Met) in the GTPBP3 gene, as well as c.752C>T (p.Thr251Ile) and c.1760C>T (p.Pro587Leu) in the POLG gene. Family segregation analysis showed that the patient's mother is a carrier of variant c.181G>C and the patient's paternal grandmother is a carrier of variant c.1199C>T in the GTPBP3 gene, which is in accordance with an autosomal recessive model of inheritance of the disease. Both variants in the POLG are found paternally inherited in the patient's healthy half-brother, thus are not considered disease-causing. GTPBP3 variants have been reported in patients with hypertrophic cardiomyopathy, associated with combined oxidative phosphorylation deficiency 23. These novel variants represent the probable cause of the observed clinical symptoms in the patient.

Impact of manual thrombectomy on microvascular obstruction in STEMI patients.

OBJECTIVE: To assess the effect of manual thrombectomy (MT) on microvascular obstruction (MVO) using cardiac magnetic resonance (CMR) in ST-segment elevation myocardial infarction (STEMI) patients undergoing primary percutaneous coronary intervention (PCI). METHODS AND RESULTS: Three hundred and eighty-three patients admitted for STEMI and undergoing CMR fulfilled the inclusion criteria and were categorized into two groups (did or did not undergo MT). The two primary endpoints were the occurrence and extent of MVO, analyzed as a categorical variable and as a semicontinuous variable. Among the 383 patients, 49.1% exhibited MVO. Both the incidence of MVO and the median number of segments presenting with MVO were significantly higher in the MT group than in the no-MT group, (59.5 vs. 38.9%, p < .001) and (1.5 [0;4] vs. 0 [0;2], p < .001). Analysis stratified on coronary thrombus grade showed similar results, only in patients with a high thrombus burden (60.7 vs. 43.5%, p = .004, and 2 [0;4] vs. 0 [0;3], p = .001. When adjusting for baseline differences, MT remained a determinant of MVO occurrence and extent (odds ratio, OR 1.802 [95% confidence interval, CI 1.080-3.009], p = .024) and ß = .137, p = .024) in patients with a high thrombus grade. CONCLUSION: In STEMI patients, MT was associated with the occurrence and extent of MVO, on CMR, especially in patients with a high thrombus burden.

Local Deuteration Enables NMR Observation of Methyl Groups in Proteins from Eukaryotic and Cell-Free Expression Systems.

Therapeutically relevant proteins such as GPCRs, antibodies and kinases face clear limitations in NMR studies due to the challenges in site-specific isotope labeling and deuteration in eukaryotic expression systems. Here we describe an efficient and simple method to observe the methyl groups of leucine residues in proteins expressed in bacterial, eukaryotic or cell-free expression systems without modification of the expression protocol. The method relies on simple stereo-selective 13 C-labeling and deuteration of leucine that alleviates the need for additional deuteration of the protein. The spectroscopic benefits of "local" deuteration are examined in detail through Forbidden Coherence Transfer (FCT) experiments and simulations. The utility of this labeling method is demonstrated in the cell-free synthesis of bacteriorhodopsin and in the insect-cell expression of the RRM2 domain of human RBM39.

NPM1 Gene Type A Mutation in Bulgarian Adults with Acute Myeloid Leukemia: A Single-Institution Study.

OBJECTIVE: Mutations of the nucleophosmin (NPM1) gene are considered as the most frequent acute myeloid leukemia (AML)-associated genetic lesion, reported with various incidences in different studies, and type A (NPM1-A) is the most frequent type. However, since most series in the literature report on the features of all patients regardless of the type of mutation, NPM1-A(+) cases have not been well characterized yet. Therefore, we evaluated the prevalence of NPM1-A in Bulgarian AML patients and searched for an association with clinical and laboratory features. MATERIALS AND METHODS: One hundred and four adults (51 men, 53 women) were included in the study. NPM1-A status was determined using allele-specific reverse-transcription polymerase chain reaction with co-amplification of NPM1-A and ß-actin and real-time quantitative TaqMan-based polymerase chain reaction. Patients received conventional induction chemotherapy and were followed for 13.2±16.4 months. RESULTS: NPM1-A was detected in 26 (24.8%) patients. NPM1-A mutation was detected in all AML categories, including in one patient with RUNX1-RUNX1T1. There were no differences associated with the NPM1-A status with respect to age, sex, hemoglobin, platelet counts, percentage of bone marrow blasts, splenomegaly, complete remission rates, and overall survival. NPM1-A(+) patients, compared to NPM1-A(-) patients, were characterized by higher leukocyte counts [(75.4±81.9)x109/L vs. (42.5±65.9)x109/L; p=0.049], higher frequency of normal karyotype [14/18 (77.8%) vs. 26/62 (41.9%); p=0.014], higher frequency of FLT3-ITD [11/26 (42.3%) vs. 8/77 (10.4%); p=0.001], and lower incidence of CD34(+) [6/21 (28.8%) vs. 28/45 (62.2%); p=0.017]. Within the FLT3-ITD(-) group, the median overall survival of NPM1-A(-) patients was 14 months, while NPM1-A(+) patients did not reach the median (p=0.10). CONCLUSION: The prevalence of NPM1-A mutation in adult Bulgarian AML patients was similar to that reported in other studies. NPM1-A(+) patients were characterized by higher leukocyte counts, higher frequency of normal karyotypes and FLT3-ITD, and lower incidence of CD34(+), supporting the idea that the specific features of type A mutations might contribute to the general clinical and laboratory profile of NPM1(+) AML patients.

And don't forget your anti in-stent stenosis pill! Commentary on 'Factors associated with in-stent stenosis after cerebral aneurysm embolization using a Pipeline embolization device' by Flores-Milan et al.

In their INR study, Flores-Milan et al. present a retrospective single-centre study that aimed to investigate and determine some of the factors associated with in-stent stenosis (ISS) after intracranial aneurysm (IA) embolization using a commercially available flow diverter stent (FD). The retrospective analyses included ruptured and unruptured intracranial aneurysms treated with standalone flow diverter stent implantation or initial coil obliteration with the FD device placed subsequently two weeks after initial treatment. The article's methodology was carefully tailored to demystify the unknown pathophysiological mechanism behind the entity of interest called in-stent stenosis. Study outcomes also included angiographic evaluation of aneurysm occlusion thrombotic and hemorrhagic events. The authors reported excellent technical and clinical results altogether. The achieved angiographic occlusion rates resonate with the current obliteration results reported in the literature. Mortality and morbidity are congruent with previously published results and were 5.3% and 1.1%, respectively.

Endovascular iliac vein recanalization for permanent pacemaker implantation in a patient who has long-term haemodialysis: a case report.

BACKGROUND: In chronic haemodialysis patients central veins occlusion occur very often. In such patients, permanent pacemaker placement implantation can be challenging and alternative approaches should be used. CASE SUMMARY: This is a case of 66-year-old male patient with complete atrioventricular block after a mitral valve (MV) surgery for endocarditis. The patient has a permanent surgically inserted haemodialysis catheter in right heart atrium after several unsuccessful attempts of endovascular recanalization of superior vena cava. A lead was implanted in the right ventricle after successful endovascular revascularization of the right iliac vein. The pacemaker was placed in a pouch on the right lower abdominal wall. DISCUSSION: To our knowledge, this is the first reported case where a permanent single-chamber pacemaker was implanted through the right iliac vein after successful endovascular recanalization in chronic haemodialysis patient post-MV replacement.

Comparison of coronary angiography and intracoronary imaging with fractional flow reserve for coronary artery disease evaluation: An anatomical-functional mismatch.

Myocardial ischemia is a leading cause of death worldwide, and it corresponds to the imbalance between blood supply and myocardial demand. Epicardial coronary artery disease (CAD) is detected on the basis of coronary angiogram, whereas invasive detection of myocardial ischemia induced by coronary stenosis is commonly based on fractional flow reserve (FFR). The use of FFR for revascularization decision-making demonstrated clinical benefit and cost-effectiveness compared with that of angiographic indices. Discrepancies between anatomical metrics and physiological assessment of CAD are frequent, which lead to change in revascularization decision from angiography compared to functional evaluation of CAD. Despite several clinical studies and guidelines recommending with high level of evidence demonstrating that FFR should be adopted in stable CAD, revascularization decision-making is still based on coronary angiogram in current practice. Because of the unique coronary anatomy, coronary stenosis characteristics, risk factors profile, and microcirculation quality, the unique evaluation based on epicardial coronary stenosis threshold failed to be a landmark of ischemia compared with FFR. Furthermore, coronary angiogram can detect only epicardial vessels, which represent only 10% of the entire coronary vasculature; therefore, microcirculation is not seen and is poorly assessed in clinical practice. Thus, the role of microcirculation is of importance in myocardial ischemia and might impact these discrepancies between angiography and FFR evaluation of CAD. In this review, we aimed to describe the poor correlation between anatomical evaluation compared with physiological evaluation to detect myocardial ischemia induced by coronary stenosis as well as the clinical implications of this visual-functional mismatch.

Direct Visualization of a Polariton Resonator in the THz Regime.

We report fabrication of a THz phonon-polariton resonator in a single crystal of LiNbO3 using femtosecond laser machining with high energy pulses. Fundamental and overtone resonator modes are excited selectively and monitored through spatiotemporal imaging. The resonator is integrated into a single solid-state platform that can include THz generation, manipulation, readout and other functionalities.

Polyphosphoester conjugates of dinuclear platinum complex: synthesis and evaluation of cytotoxic and the proapoptotic activity.

Macromolecular conjugates of a dinuclear platinum complex with a spermidine bridge were synthesized using poly(oxyethylene H-phosphonate)s as precursor polymer. The complex species were attached to the polymer chain via a phosphoramide bond resulting from the reaction between the H-phosphonate groups and the middle amino group of the spermidine moiety. (1)H and (31)P{H} DOSY NMR spectral data were used to prove the conjugation reaction and to characterize the new species. The conjugates exhibited profound cytotoxicity in a panel of five chemosensitive human tumor cell lines and one cisplatin-resistant model (HL-60/CDDP), and were found to induce apoptotic cell death. A flow cytometric analysis encountered a cisplatin-dissimilar modulation of the cell cycle progression in KG-1 leukemic cells, following exposure to the dinuclear agents. Moreover, the novel compounds displayed less pronounced inhibitory activity against cultured murine renal epithelial cells, as compared to cisplatin.

Apoptotic mechanisms of the biotechnologically produced arylnaphtalene lignan justicidin B in the acute myeloid leukemia-derived cell line HL-60.

BACKGROUND: The present study aimed at optimization of the biotechnological production of the lignan justicidin B by genetically transformed cultures of Linum leonii and the pharmacological evaluation of the pro-apoptotic effects of the compound in HL-60 cells. METHODS: A rapidly growing selected root line of L. leonii was grown in 2-L bioreactor for period of 40 days and the protocols for obtaining of the compound have been optimized. The pharmacological study included evaluation of the cytotoxicity of the compound in HL-60 cells (MTT-assay), its apoptogenic effects and its effects on caspase 3,8 and 9 activation. RESULTS: After 40 days of sterile run scale up of hairy root culture in bioreactor, 27.2g/L dry weight of root biomass was harvested from the bioreactor culture vessel, recording about nine times increase over initial inoculum (3.0g), with 1.55%±0.07 Justicidin B, greater than yields from 300ml flasks. Our findings are the first work toward the scale up of L. leonii hairy roots-based biotechnological production of Justicidin B, employing bioreactors for high biomass production to meet the industrial requirement. The results from the pharmacological evaluation have shown that the tested arylnaphtalene lignan is a potent cytotoxic and proapoptotic agent against HL-60. The induction of apoptosis proceeds via activation of the intrinsic mitochondrial cell-death signaling pathways. CONCLUSION: The potent activity at low micromolar concentration and the feasibility of biotechnological production of justicidin B implies that there is enormous scope in its further evaluation as possible antineoplastic drug candidate.

A particular expression pattern of CD13 epitope 7H5 in chronic lymphocytic leukaemia--a possible new therapeutic target.

A total of 50 patients with chronic lymphocytic leukaemia (CLL), as well as the B-cell leukaemia cell lines MEC-1, JVM-3, and BV-173 were studied in order to assess the incidence of CD13/aminopeptidase N (APN) immunolabelling with a monoclonal antibody 7H5 compared to LeuM7 and to CD13 mRNA levels, and to correlate these data with the cytotoxic and apoptosis-induction activity of the natural phenolic APN inhibitor curcumin. CD13/APN was detected in a significant proportion of B-CLL patients (42/50, 84%), immunolabelled by 7H5 (42/50) ± LeuM7 (10/50). Molecular analysis for CD13 transcripts confirmed these data, resulting in a specific RT-PCR product in CD13 positive cases. Curcumin showed concentration-dependent cytoreductive efficacy and apoptosis-induction activity in all tested cell lines and primary cultures from CLL mononuclear cells. There was a clear tendency for a better response in CD13 positive cases. The incidence of CD13/APN in CLL suggests that the inhibition of APN/CD13 by curcumin may be an effective new molecular target for a more efficient therapy for these patients and warrants further investigations.

Chronic eosinophilic leukemia with FIP1L1-PDGFRA transcripts after occupational and therapeutic exposure to radiation.

We present for the first time a 40-year-old male patient with a 20 year history of occupational exposure to radiation as a nuclear power plant worker, who developed FIP1L1-PDGFRA-positive chronic eosinophilic leukemia 27 months after radiotherapy for testicular seminoma. After an one-year history of dry cough, itching and night sweats, the patient presented with an elevated leukocyte count with absolute eosinophilia of 14.2×10(9)/L, bone marrow and lymph node involvement. Treatment with Imatinib was initiated, resulting in complete hematological remission at the sixth month and complete molecular response by nested primers reverse transcription polymerase chain reaction - at the end of the first year. This case contributes to the clinical heterogeneity of a rare entity such as FIP1L1-PDGFA-positive myeloproliferative neoplasms, and for the possible role of occupational and therapeutic radiation, raising the question if one or both of them might be the causative factor.

Terahertz polariton propagation in patterned materials.

Generation and control of pulsed terahertz-frequency radiation have received extensive attention, with applications in terahertz spectroscopy, imaging and ultrahigh-bandwidth electro-optic signal processing. Terahertz 'polaritonics', in which terahertz lattice waves called phonon-polaritons are generated, manipulated and visualized with femtosecond optical pulses, offers prospects for an integrated solid-state platform for terahertz signal generation and guidance. Here, we extend terahertz polaritonics methods to patterned structures. We demonstrate femtosecond laser fabrication of polaritonic waveguide structures in lithium tantalate and lithium niobate crystals, and illustrate polariton focusing into, and propagation within, the fabricated waveguide structures. We also demonstrate a 90 degrees turn within a structure consisting of two waveguides and a reflecting face, as well as a structure consisting of splitting and recombining elements that can be used as a terahertz Mach-Zehnder interferometer. The structures permit integrated terahertz signal generation, propagation through waveguide-based devices, and readout within a single solid-state platform.

Direct visualization of the Gouy phase by focusing phonon polaritons.

We report the generation of aberration-free cylindrical phonon-polariton wave packets in uniaxial LiTaO3 crystals by nonresonant impulsive stimulated Raman scattering. The unique properties of phonon polaritons with a typical carrier frequency in the THz regime allow direct measurement of the spatiotemporal amplitude and phase distributions. We demonstrate that under these conditions the phase anomaly (Gouy phase) may be visualized directly through spatiotemporal imaging as the cylindrical wave propagates through its focus.

Coherent control of phonon-polaritons in a terahertz resonator fabricated with femtosecond laser machining.

Using femtosecond laser machining, we fabricated a terahertz resonant cavity in LiNbO3. Optical pulse sequences with variable repetition rates, generated through a novel pulse-shaping method, are used for characterization of the cavity resonances and for amplification of terahertz phonon-polaritons in the cavity.