Gradual Shortening Research Articles

Individually Rate Corrected QTc Intervals in Children and Adolescents

Accurate evaluation of the appearance of QTc sex differences during childhood and adolescence is intricate. Inter-subject differences of individual QT/RR patterns make generic heart rate corrections inaccurate because of fast resting heart rates in children. The study investigated 527 healthy children and adolescents aged 4–19 years (268 females, 50.9%). All underwent continuous ECG 12-lead monitoring while performing postural changes during a 70-min investigative protocol to obtain QT interval measurements at different heart rates. On average, more than 1200 ECG measurements (QT interval and its 5-min history of preceding RR intervals) were made in each subject. Curvilinear QT/RR regression involving intra-individual correction for QT/RR hysteresis were calculated in each subject. The projection of the QT/RR regressions to the heart rate of 60 beats per minute defined individually corrected QTc intervals. In males, gradual QTc shortening by about 15 ms appeared during the ages of 13–19 years synchronously with the incidence of secondary sex signs (p = 0.016). On the contrary, whilst gradual QTc prolongation by about 10 ms appeared in females, it occurred only during ages 16–19 years and was not related to the incidence of secondary sex signs (p = 0.18). The study also showed that in children and adolescents, linear QT/RR models fit the intra-subject data significantly more closely than the log-linear models (p < 0.001). The study speculates that hormonal shifts during puberty might be directly responsible for the QTc shortening in males but that QTc prolongation in females is likely more complex since it was noted to follow the appearance of secondary sex signs only after a considerable delay.

Occupational inhalation poisoning with the veterinary antibiotic tiamulin

Introduction: Tiamulin is a semisynthetic pleuromutilin diterpene veterinary antibiotic, widely used in farms. We present a case of prolonged QT-interval and ventricular tachyarrhythmia after tiamulin inhalation.Case presentation: A 43-year-old veterinarian without previous medical history was dividing granulated powder of antibiotic gravimetrically without wearing personal protective equipment. Half an hour after exposure, nausea occurred; four hours later he started to vomit and soon after that he experienced syncope. He was unconscious three minutes; afterwards he became somnolent, dizzy and nauseated with sweating and salivation. On admission to hospital five hours after exposure, he was conscious and had heart rate 70 beats/min and blood pressure 140/80 mmHg. Initial laboratory results were normal. Electrocardiography showed a prolonged QTc-interval of 730 ms with numerous polymorphic ventricular extrasystoles and episodes of non-sustained polymorphic ventricular tachycardia that resolved after treatment with lidocaine and magnesium. Subsequent electrocardiography revealed gradual shortening of QTc-interval with QTc-interval normalization (430 ms) between 24 and 32 hours after tiamulin exposure. Laboratory tests, morphologic heart diagnostics and genetic testing excluded other potential causes of QTc-interval prolongation. Subsequent toxicology analysis by LC-MS/MS confirmed tiamulin in his serum samples on admittance (500 ng/mL).Conclusion: Tiamulin inhalation can be associated with prolonged QT-interval and ventricular tachyarrhythmia. QT-interval prolongation could be expected in overdoses of emerging human pleuromutilins.

The Evolution of the Maxillary Canal in Probainognathia (Cynodontia, Synapsida): Reassessment of the Homology of the Infraorbital Foramen in Mammalian Ancestors

In mammals, the infraorbital canal provides a passage for the infraorbital ramus of the maxillary branch of the trigeminal nerve. The infraorbital nerve ensures tactile sensitivity of the upper teeth and face between the eye and upper lip and, more significantly, the innervation of mystacial vibrissae (whiskers). In contrast, most non-mammalian synapsids display a more “reptilian-like” ancestral condition in which a long and ramified maxillary canal completely enclosed the infraorbital nerve along with other branches of the trigeminal nerve. The phylogenetic transition from the ancestral “reptilian-like” to the derived “mammal-like” condition has been hypothesized to occur at the base of the Probainognathia clade. Using μCT and synchrotron scanning, this study aims to document this transition in detail by focusing on a sample of non-mammalian probainognathian cynodonts and early mammaliaforms. We find that the mammalian condition is the result of a gradual shortening of the maxillary canal, which enabled the infraorbital nerve to ramify within the soft tissues of the face. Mobile whiskers became possible only after the mammalian infraorbital nerve had evolved, which suggest that these structures appeared in Probainognathus and more derived cynodonts. Finally a foramen located on the ventral margin of the lacrimal bone, which has been often homologized with the infraorbital foramen of derived Probainognathia and early Mammaliaformes, is most probably homologous to the mammalian zygomaticofacial foramen.

Abstract 1407: Telomere length and melanoma survival

Abstract Telomeres at chromosomal ends are comprised of multiple short repeat sequences. In humans TTAGGG repeats account for 10-15 kb of double-stranded telomeres that end in single stranded G-rich tails of about 150-200 nucleotides. Telomeres in somatic cells undergo gradual shortening due to inherent limitations of DNA replication and limited levels of specialized enzyme telomerase. We previously reported an association between increased telomere length and risk of melanoma. In this study based on 1019 stage I and II cutaneous melanoma patients, we determined the effect of telomere length on patient survival. The telomere length was measured in DNA from peripheral blood cells using monochrome multiplex quantitative real-time PCR where telomere repeat copy number (T) was compared to a single-copy gene, albumin (S) and results presented as relative telomere length based on T/S ratio. A univariate Cox regression showed increased patient survival with increased telomere length (HR 0.65, 95% CI 0.42-1.00, P 0.05). With every decrease in 1 unit of telomere length the estimated odds of survival decreased by an approximate factor of 1.54 (95%CI 1.00-2.38). A model based on median distribution showed short rather than long telomere associated with poor survival with hazard ratio (HR) of 2.05 (95% confidence interval (CI) 1.33-3.16; log rank P 0.001); the model after adjustment with confounding factors including age at diagnosis showed a HR of 1.50 (95%CI 0.96-2.34). Stratification of patients based on median age showed statistically significant effect of decreased telomere length on poor survival in patients below the median age (HR 2.27 95%CI 1.09-4.27) but not in patients above the median age (HR 1.42 95%CI 0.82-2.46). Distribution of the patients into quantiles based on age showed the largest effect of decreased telomere length in patients younger than 30 years (HR 3.90 95%CI 1.28-11.93); the effect decreased with increasing age. Three single nucleotide polymorphisms, rs131178082, rs7726159 and rs6060627, previously shown to be associated with telomere length, were used to construct a weighted genetic score and paradoxically, the patients carrying the alleles associated with increased telomere length showed poor survival with shorter telomeres (HR 2.94 95%CI 1.54-5.62). Multivariate analysis that included, age, sex, outdoor life style, presence of actinic keratosis, number of nevi, tumor location, tumor stage, Breslow thickness and tumor ulceration showed a statistically significant association between composite genetic score and poor survival (HR 2.68 95%CI 1.24-5.80). Thus, the results from the study show that in contrast to association between melanoma risk and increased telomere length, the decreased telomere length associates with poor patient survival. The adverse effect on survival due to decreased telomere length was more pronounced in young rather than old patients and on the patients carrying genotypes for longer telomeres. Citation Format: Sivaramakrishna Rachakonda, Nalini Srinivas, Zaida Garcia-Casado, Celia Requena, Eduardo Nagore, Rajiv Kumar. Telomere length and melanoma survival [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 1407.

Effects of antiarrhythmics and hypokalemia on the rate adaptation of cardiac repolarization

Objectives. In normal conditions, sudden heart rate acceleration provokes a rapid reduction in ventricular action potential duration (APD). The protracted APD rate adaptation favors early afterdepolarizations and precipitates arrhythmia. Nevertheless, it is uncertain as to whether the rate-dependent changes of ventricular repolarization can be adversely modified by arrhythmogenic drugs (quinidine and procainamide) and hypokalemia, in comparison to the agents with safe therapeutic profile, such as lidocaine. Design. The rate adaptation of QT interval and monophasic APD obtained from the left ventricular (LV) and the right ventricular (RV) epicardium was examined during rapid cardiac pacing applied in isolated, perfused guinea-pig heart preparations. Results. At baseline, an abrupt increase in cardiac activation rate was associated with a substantial reduction of the QT interval and ventricular APD in the first two cardiac cycles, which was followed by a gradual shortening of repolarization over subsequent pacing intervals. The time constants of the fast (τfast) and slow (τslow) components of the APD dynamics determined from a double exponential fit were longer in RV compared to LV chamber. Quinidine, procainamide, and hypokalemia prolonged ventricular repolarization and delayed the rate adaptation of the QT interval and APD in LV and RV, as evidenced by increased τfast and τslow values. In contrast, lidocaine had no effect on the dynamic changes of ventricular repolarization upon heart rate acceleration. Conclusions. The rate adaptation of ventricular repolarization is delayed by arrhythmogenic interventions, such as quinidine, procainamide, and hypokalemia, but not changed by lidocaine, a clinically safe antiarrhythmic agent.

Linagliptin prevents atrial electrical and structural remodeling in a canine model of atrial fibrillation.

Dipeptidyl peptidase 4 (DPP-4) inhibitors have recently been reported to exhibit additional cardioprotective effects; however, their effect in atrial remodeling, such as in atrial fibrillation (AF), remains unclear. In this study, the effect of linagliptin on atrial electrical and structural remodeling was evaluated in a canine AF model. Sixteen beagle dogs with 3-week atrial rapid stimulation were divided into the linagliptin group (9mg/kg/day, n = 8) and pacing control group (n = 8). Three additional dogs without rapid pacing were assigned into non-pacing group, which was used as sham in this study. In the dogs with rapid pacing, the atrial effective refractory period (AERP), conduction velocity (CV), and AF inducibility were evaluated and blood was sampled every week. After the entire protocol, atrial tissue was sampled for histological examinations using HE, Azan, and dihydroethidium (DHE) staining to evaluate any tissue damage or oxidative stress. The pacing control group exhibited a gradual AERP shortening and CV decrease along the time course as previously reported. In the linagliptin group, the AERP shortening was not affected, but the CV decrease was suppressed in comparison to the control group (p < 0.05). The AF inducibility was increased in the control group and suppressed in the linagliptin group (p < 0.05). The control group exhibited tissue fibrosis, the degree of which was suppressed in the linagliptin group. DHE staining exhibited suppression of the reactive oxygen species expression in the linagliptin group in comparison to the pacing control group. Linagliptin, a DPP-4-inhibitor, suppressed the AF inducibility, CV decrease, and overexpression of oxidative stress in the canine AF model. Such suppressive effects of linagliptin on AF in the canine model may possibly be related to the anti-oxidative effect.

Glucose Responsive Rheological Change and Drug Release from a Novel Worm-like Micelle Gel Formed in Cetyltrimethylammonium Bromide/Phenylboronic Acid/Water System.

A novel glucose (Glc)-responsive gel formed by worm-like micelles (WLMs) has the potential to provide a self-regulating insulin delivery system. We have prepared a WLM gel system using 75 mM cetyltrimethylammonium bromide, 75 mM phenylboronic acid, and water. At pH 9.4, this gel-like system was highly viscous and supported its own weight, and dynamic viscoelasticity measurement indicated that it contained long and entangled WLMs. The visual observation of gels prepared to include >6 mM Glc revealed that these adopted a sol-like appearance, whereas those prepared to include a control compound (2-10 mM diethylene glycol) retained their gel-like appearance. The storage modulus ( G') of this system decreased as the Glc concentration increased (2-10 mM), indicating a gradual shortening of the WLMs. In vitro release was evaluated using a test compound (fluorescein isothiocyanate dextran) in a microsized flow system. By 120 min, the release of this compound from the WLM gel was around 27-fold greater in the presence of 100 mM Glc than without Glc or with 100 mM diethylene glycol. This demonstrated the successful preparation of a WLM gel that showed an altered drug release rate, depending on Glc concentration.

Use of U-STELA for Accurate Measurement of Extremely Short Telomeres.

Telomeres are repetitive genetic materials that protect the chromosomes by capping the ends of chromosomes. Each time a cell divides, telomeres get shorter. Telomere length is mainly maintained by telomerase. This enzyme is present in the embryonic stem cells in high concentrations and declines with age. It is still unclear to what extend there is telomerase in adult stem cells, but considering these are the founder cells to the cells of the all tissues in a body, understanding the telomere dynamics and expression of telomerase in adult stem cells is very important.Telomere length has been implicated as one of the markers for neoplastic transformation in both in vivo and in vitro studies. During cancerogenesis, telomeres shorten due to high cell turnover and repeats are added by active telomerase or alternative lengthening of telomeres (ALT). This gradual shortening is replication driven and does not necessarily explain the presence of ultrashort telomeres. Ultrashort telomeres are observed when there is a sudden shortening in telomeres not related with cell division and may arise from breaks in telomeres due to oxidative damage and replication slippage.Universal STELA is an accurate method for evaluation of ultrashort telomeres in hMSC-telo1 cells. Compared to TRF assay, U-STELA is developed to overcome several problems in detecting abrupt telomere shortening in a single chromosome.

Abstract 3408: Telomere length and TERT promoter mutations in cutaneous melanoma

Abstract Telomeres at chromosomal ends are comprised of multiple short repeat sequences. In humans TTAGGG repeats account for telomere length ranging 10-15 kb. Telomere sequences are mainly double stranded that end in a single stranded G-rich tail of 150-200 nucleotides. Telomeres in somatic cells undergo gradual shortening due to inherent limitations of DNA replication and limited levels of specialized enzyme telomerase that adds the repeats at chromosomal ends to maintain homeostasis. We previously reported somatic mutations in the core promoter of the telomerase reverse transcriptase (TERT) gene that lead to increased transcription of catalytic subunit and tumor specific telomerase reactivation. Telomere length per se is associated with risk in different cancers. In this study, we measured leukocyte telomere length using real-time PCR in 1469 melanoma patients and compared with that in 1158 matched healthy controls. The melanoma patients had statistically significantly longer telomeres than matched controls (t-test; P 6X10^-10). Mendelian randomization, carried out using two polymorphisms represented by rs1317082 and rs7726159 that associated with telomere length in genome wide association studies, showed association between increased telomere length and melanoma risk with an odds ratio of 2.3 (95% confidence interval 1.8-2.8). When measured in blood tissues from a melanoma family with the germline -57A&amp;gt;C TERT promoter mutation, the carriers had longer telomeres (median 1.12) than the non-carriers (median 0.87). The TERT promoter mutations create binding motifs for E-twenty six (ETS) transcription factors and in stem cell the presence of the promoter mutations resulted in continued TERT expression and telomerase activity following differentiation into adult cells. Individuals with the germline TERT promoter mutations develop melanoma with an early age of onset and rapid progression to metastases; two individuals in the family who lived past median age of onset developed several other malignancies, besides melanoma. Thus, dysregulated telomerase leads to a severe phenotype. In an analogy, we hypothesize that association of longer rather than shorter telomeres with an increased risk of melanoma reflects stochastic increased telomerase levels due to common genetic variation. In contrast, the telomere length was shorter in tumors from unrelated melanoma patients with (121) than without (170) somatic TERT promoter mutations (P 1X10^-5), which reflects the selection of the mutations at telomere crisis. Thus, a dynamic but controlled system evolved around telomere homeostasis when dysregulated leads to an increased cancer risk and affects tumor progression. Citation Format: Sivaramakirishna Rachakonda, Barbara Heidenreich, Eduardo Nagore, Rajiv Kumar. Telomere length and TERT promoter mutations in cutaneous melanoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 3408. doi:10.1158/1538-7445.AM2017-3408

TZAP-ing telomeres down to size.

The phenomenon of gradual telomere shortening has become a paradigm for how we understand the biology of aging and cancer. Cell proliferation is accompanied by cumulative telomere loss, and the aged cell either senesces, dies or transforms toward cancer. This transformation requires the activation of telomere elongation mechanisms in order to restore telomere length such that cell death or senescence programs are not induced. Most of the time, this occurs through telomerase reactivation. In other rare cases, the Alternative lengthening of telomeres (ALT) pathway hijacks DNA recombination-associated mechanisms to hyperextend telomeres, often to more than 50 kb. Why telomere length is restricted and what sets their maximal length has been a long-standing puzzle in cell biology. Two recent studies published in this issue of EMBO Reports [1] and recently in Science [2] sought to address this important question. Both built on omics approaches that identified ZBTB48 as a potential telomere-associated protein and reveal it to be a critical regulator of telomere length homeostasis by the telomere trimming mechanism. These discoveries provide fundamental insights for our understanding of telomere trimming and how it impacts telomere integrity in stem and cancer cells.

CTC1-mediated C-strand fill-in is an essential step in telomere length maintenance.

To prevent progressive telomere shortening as a result of conventional DNA replication, new telomeric DNA must be added onto the chromosome end. The de novo DNA synthesis involves elongation of the G-rich strand of the telomere by telomerase. In human cells, the CST complex (CTC1-STN1-TEN1) also functions in telomere replication. CST first aids in duplication of the telomeric dsDNA. Then after telomerase has extended the G-rich strand, CST facilitates fill-in synthesis of the complementary C-strand. Here, we analyze telomere structure after disruption of human CTC1 and demonstrate that functional CST is essential for telomere length maintenance due to its role in mediating C-strand fill-in. Removal of CTC1 results in elongation of the 3΄ overhang on the G-rich strand. This leads to accumulation of RPA and telomeric DNA damage signaling. G-overhang length increases with time after CTC1 disruption and at early times net G-strand growth is apparent, indicating telomerase-mediated G-strand extension. In contrast, C-strand length decreases continuously, indicating a deficiency in C-strand fill-in synthesis. The lack of C-strand maintenance leads to gradual shortening of the telomeric dsDNA, similar to that observed in cells lacking telomerase. Thus, telomerase-mediated G-strand extension and CST-mediated C-strand fill-in are equally important for telomere length maintenance.

How reversible is development? Contrast between developmentally plastic gain and loss of segments in barnacle feeding legs.

Segmented organisms and structures have fascinated biologists since William Bateson first described homeotic transformation and recognized the fundamental evolutionary significance of segmental organization. On evolutionary time scales, segments may be lost or gained during major morphological transitions. But how segment loss compares to gain on developmental time scales remains mysterious. Here, we examine the ease of reverse development (opposite to normal growth) by comparing developmentally plastic leg segment loss versus gain in individual barnacles transplanted between different water flow conditions. Plastic segment addition occurred rapidly (one to two molts) and exclusively near the limb base. In contrast, developmentally plastic segment loss-the first observation in any arthropod-took much longer (>10 molts) and, remarkably, occurred throughout the leg (23% of losses occurred mid-limb). Segment loss was not a simple reversal of segment addition. Intersegmental membranes fused first, followed by elimination of duplicate tendons and gradual shortening (but not loss) of duplicate setae. Setal loss, in particular, may impose a severe developmental constraint on arthropod segment fusion. This asymmetric developmental potential (time lag of phenotypic response)-plastic segment addition (amplified normal development) is faster and more orderly than segment loss (reverse development)-adds a new dimension to models of developmental plasticity because the cost of making a developmental mistake in one direction will be greater than in the other.

Solar-driven advanced oxidation processes for full mineralisation of azo dyes in wastewater

Environmental contextFull mineralisation of synthetic azo dyes in industrial wastewater is a tough job for traditional wastewater treatment technologies. There is an urgent need for the development of both sustainable and environmentally friendly technology capable of fully mineralising these azo compounds. We show that solar-driven advanced oxidation processes are capable of completely mineralising azo compounds with high utilisation of solar energy. AbstractMineralisation of synthetic azo dyes in industrial wastewater is an energy-intensive process in treatment technology. The Solar Thermal Electrochemical Process for advanced oxidation processes (STEP-AOPs) utilises solar energy and electricity for the activation and electrooxidation of organic pollutants to harmless, small and non-toxic molecules with no other energy consumption. Based on molecular structure and chemistry, the STEP-AOPs for the treatment of azo dyes in wastewater, as exemplified with a typical azo dye, methyl orange, is reported for the first time. Thermodynamic calculations of the temperature-dependent potentials of methyl orange demonstrate that Gibbs free energy decreased by 161kJmol–1 and the potential decreased by 0.019V with an increase of temperature from 20 to 80°C, which indicates that the drop in both energy and potential specifically fits the STEP-AOPs technique. Experimental results showed that the STEP-AOPs achieved a total organic carbon (TOC) removal of 95.6% for methyl orange. The TOC removal rate improved by 39.8% and the unit TOC electricity consumption decreased by 53.8% at 80°C compared with conventional methods (20°C). The mineralisation mechanism for methyl orange was a gradual shortening of the molecular chain through cleavage of the azo bond, breakdown of the benzene ring and formation of inorganic small molecules susceptible to be oxidised to non-toxic small molecules, and carbon dioxide via STEP-AOPs. The evidence shows that the STEP-AOPs is capable of mineralising azo compounds completely.

Fission yeast Stn1 is crucial for semi-conservative replication at telomeres and subtelomeres.

The CST complex is a phylogenetically conserved protein complex consisting of CTC1/Cdc13, Stn1 and Ten1 that protects telomeres on linear chromosomes. Deletion of the fission yeast homologs stn1 and ten1 results in complete telomere loss; however, the precise function of Stn1 is still largely unknown. Here, we have isolated a high-temperature sensitive stn1 allele (termed stn1-1). stn1-1 cells abruptly lost telomeric sequence almost completely at the restrictive temperature. The loss of chromosomal DNA happened without gradual telomere shortening, and extended to 30 kb from the ends of chromosomes. We found transient and modest single-stranded G-strand exposure, but did not find any evidence of checkpoint activation in stn1-1 at the restrictive temperature. When we probed neutral-neutral 2D gels for subtelomere regions, we found no Y-arc-shaped replication intermediates in cycling cells. We conclude that the loss of telomere and subtelomere DNAs in stn1-1 cells at the restrictive temperature is caused by very frequent replication fork collapses specifically in subtelomere regions. Furthermore, we identified two independent suppressor mutants of the high-temperature sensitivity of stn1-1: a multi-copy form of pmt3 and a deletion of rif1. Collectively, we propose that fission yeast Stn1 primarily safeguards the semi-conservative DNA replication at telomeres and subtelomeres.

TERT promoter mutations in telomere biology.

Telomere repeats at chromosomal ends, critical to genome integrity, are maintained through an elaborate network of proteins and pathways. Shelterin complex proteins shield telomeres from induction of DNA damage response to overcome end protection problem. A specialized ribonucleic protein, telomerase, maintains telomere homeostasis through repeat addition to counter intrinsic shortcomings of DNA replication that leads to gradual sequence shortening in successive mitoses. The biogenesis and recruitment of telomerase composed of telomerase reverse transcriptase (TERT) subunit and an RNA component, takes place through the intricate machinery that involves an elaborate number of molecules. The synthesis of telomeres remains a controlled and limited process. Inherited mutations in the molecules involved in the process directly or indirectly cause telomeropathies. Telomerase, while present in stem cells, is deactivated due to epigenetic silencing of the rate-limiting TERT upon differentiation in most of somatic cells with a few exceptions. However, in most of the cancer cells telomerase reactivation remains a ubiquitous process and constitutes one of the major hallmarks. Discovery of mutations within the core promoter of the TERT gene that create de novo binding sites for E-twenty-six (ETS) transcription factors provided a mechanism for cancer-specific telomerase reactivation. The TERT promoter mutations occur mainly in tumors from tissues with low rates of self-renewal. In melanoma, glioma, hepatocellular carcinoma, urothelial carcinoma and others, the promoter mutations have been shown to define subsets of patients with adverse disease outcomes, associate with increased transcription of TERT, telomerase reactivation and affect telomere length; in stem cells the mutations inhibit TERT silencing following differentiation into adult cells. The TERT promoter mutations cause an epigenetic switch on the mutant allele along with recruitment of pol II following the binding of GABPA/B1 complex that leads to mono-allelic expression. Thus, the TERT promoter mutations hold potential as biomarkers as well as future therapeutic targets.

Monitoring of selected non-ionic surfactants in river water by liquid chromatography–tandem mass spectrometry

Alcohol ethoxylates (AEs) are a significant component of the non-ionic surfactant (NS) flux discharged into surface water. Due to the polydispersity of the majority of NS, they are easily recognizable by their ‘fingerprints’, i.e. a series of mass peaks which differ by m/z = 44, namely the m/z of a single oxyethylene subunit. Dodecanol ethoxylates (C12EOx) represent AEs from both renewable and petrochemical sources. Therefore, C12Ex are suitable fingerprints of NS in the aquatic environment.The aim of this work was to develop an LC–MS/MS method suitable for AE monitoring in river water.River water samples taken from the River Warta in Poznan (Poland) were extracted with ethyl acetate, evaporated, reconstituted in the mobile phase and processed by the LC – Multistage MS procedure (LC–MS/MS) using optimum multiple reaction monitoring (MRM). The method of multiple standard additions was used for the evaluation of each AE fingerprint concentration. The concentration of C12EOx having 2–9 oxyethylene subunits was determined. Standards for higher C12EOx are not yet available. The developed method offers an LOD of between 1 and 9 ng L−1, and is suitable for the monitoring of NS fingerprints in river water.The range of C12EO2–9 concentrations determined in the River Warta varied within two orders of magnitude in all cases. The lowest determined concentration was 17 ± 1 ng L−1, while the highest was 2.6 ± 0.14 μg L−1. The total concentration of C12EO2–C12EO9 homologues varied between 1.4 and 11.2 μg L−1.A relatively high concentration of short-chained homologues (2–5 oxyethylene subunits) was observed in the investigated river water. This provides evidence of a biodegradation pathway involving the gradual shortening of the AE oxyethylene chain. Distinct evidence was also obtained of unregulated NS discharges into the river.

Clinical Application and Exploration on Steps Progressively Corrections and Gradual Shortening Combined with Total Spinal Osteotomy for Angular Kyphosis—Basic Research and Clinical Application

Introduction: This study is designed to explore the biomechanical principles of posterior bilateral pedicle screw on steps progressively and tight closure on every step for severe angular kyphosis and the indication conditions and clinical pathways of above new technique. Materials and Methods: A total of 90 patients have severe angular kyphosis, 37 males and 53 females, with an average age of 47 years. All patients were treated with posterior bilateral pedicle screw and step tight closure for gradual shortening and orthopedic technology. Results: Average intraoperative blood loss was 2089 ml. Average operation time was 326 minutes. Kyphotic angle changed from 90.1° averagely preoperatively to 41.6° averagely postoperatively with an improvement rate of 65%. The distance from C7 plumb line to posterior upper edge of the S1 vertebral body was averagely 5.2 mm postoperatively, with a correction rate of 73%. Neurological complications occurred in 2 cases, accounting for 6%. Non-neurological complications occurred in 2 cases, accounting for 6%. Anatomical parameters were significantly improved postoperatively (P < 0.05). All patients were averagely followed up for 28 months. Segments undergoing osteotomy achieved bone fusion, without loss of orthopedic angle. Conclusions: The orthopedic technology of bilateral pedicle screw and step tight closure designed by human physiology and biomechanical principles can protect spinal cord cells from injury to the utmost. To choose an appropriate osteotomy plane during pre-surgical planning can make the repair results closer to the normal function of the spine. Intraoperative nerve root protection, full release and bone graft fusion are effective safeguards to ensure kyphosis correction and to avoid spine lateral offset.

The Interaction Between Heart Systole and Cerebral Circulation During Lower Body Negative Pressure Test.

The time constant (τ[s]) estimates how fast the arterial part of the cerebrovascular bed fills with blood volume during the cardiac cycle, whereas a product of τ and heart rate (HR) (τ*HR[%]) assesses how this period of arterial filling is related to an entire heart cycle. In this study we aimed to investigate cerebral hemodynamics using τ and τ*HR during a progressive lower body negative pressure (LBNP) test.Transcranial Doppler cerebral blood flow velocity (CBFV), Finapres arterial blood pressure (ABP), and HR, along with end-tidal CO2, were simultaneously recorded in 38 healthy volunteers during an LBNP test. The τ was estimated using mathematical transformation of ABP and CBFV pulse waveforms. After a gradual shortening of τ from baseline (0.20 ± 0.06 s) to maximal LBNP before the onset of presyncope (0.15 ± 0.05 s), we observed a significant increase in τ at presyncope (0.24 ± 0.15 s; p = 0.0001). In the course of LBNP, the τ*HR did not significantly change from baseline (25.6 ± 5.7 % vs 26.6 ± 8.9 %, p = n.s.) except for presyncope, when it increased to 40.4 ± 21.1 % (p < 0.000001). Because the time needed to fill the arterial part of the cerebrovascular bed with blood is prolonged during presyncope, an increased part of the heart cycle seems to be spent on the cerebral blood supply.

Rod-to-Globule Transition of pDNA/PEG-Poly(l-Lysine) Polyplex Micelles Induced by a Collapsed Balance Between DNA Rigidity and PEG Crowdedness.

The role of poly(ethylene-glycol) (PEG) in rod-shaped polyplex micelle structures, having a characteristic core of folded plasmid DNA (pDNA) and a shell of tethered PEG chains, is investigated using PEG-detachable polyplex micelles. Rod shapes undergo change to compacted globule shapes by removal of PEG from polyplex micelles prepared from block copolymer with acid-labile linkage between PEG and poly(l-lysine) (PLys) through exposure to acidic milieu. This structural change supports the previous investigation on the rod shapes that PEG shell prevents the DNA structure from being globule shaped as the most favored structure in minimizing surface area. Noteworthy, despite the PEG is continuously depleted, the structural change does not occur in gradual shortening manner but the rod shapes keep their length unchanged and abruptly transform into globule shapes. Analysis of PEG density reveals the transition occurred when tethered PEG of rod shapes has decreased to a critical crowdedness, i.e., discontacted with neighboring PEG, which eventually illuminates another contribution, rigidity of DNA packaged as bundle in the rod shapes, in addition to the steric repulsion of PEG, in sustaining rod shapes. This investigation affirms significant role of PEG and also DNA rigidity as bundle in the formation of rod-shaped structures enduring the quest of compaction of charge-neutralized DNA in the polyplex micelles.

Shortening a loop can increase protein native state entropy.

Protein loops are essential structural elements that influence not only function but also protein stability and folding rates. It was recently reported that shortening a loop in the AcP protein may increase its native state conformational entropy. This effect on the entropy of the folded state can be much larger than the lower entropic penalty of ordering a shorter loop upon folding, and can therefore result in a more pronounced stabilization than predicted by polymer model for loop closure entropy. In this study, which aims at generalizing the effect of loop length shortening on native state dynamics, we use all-atom molecular dynamics simulations to study how gradual shortening a very long or solvent-exposed loop region in four different proteins can affect their stability. For two proteins, AcP and Ubc7, we show an increase in native state entropy in addition to the known effect of the loop length on the unfolded state entropy. However, for two permutants of SH3 domain, shortening a loop results only with the expected change in the entropy of the unfolded state, which nicely reproduces the observed experimental stabilization. Here, we show that an increase in the native state entropy following loop shortening is not unique to the AcP protein, yet nor is it a general rule that applies to all proteins following the truncation of any loop. This modification of the loop length on the folded state and on the unfolded state may result with a greater effect on protein stability.