Functional Analysis of a Unique Troponin C Mutation, GLY159ASP, that Causes Familial Dilated Cardiomyopathy, Studied in Explanted Heart Muscle

Background-Familial dilated cardiomyopathy can be caused by mutations in the proteins of the muscle thin filament. In vitro, these mutations decrease Ca2+ sensitivity and cross-bridge turnover rate, but the mutations have not been investigated in human tissue. We studied the Ca2+-regulatory properties of myocytes and troponin extracted from the explanted heart of a patient with inherited dilated cardiomyopathy due to the cTnC G159D mutation.Methods and Results-Mass spectroscopy showed that the mutant cTnC was expressed approximately equimolar with wild-type cTnC. Contraction was compared in skinned ventricular myocytes from the cTnC G159D patient and nonfailing donor heart. Maximal Ca2+-activated force was similar in cTnC G159D and donor myocytes, but the Ca2+ sensitivity of cTnC G159D myocytes was higher (EC50 G159D/donor=0.60). Thin filaments reconstituted with skeletal muscle actin and human cardiac tropomyosin and troponin were studied by in vitro motility assay. Thin filaments containing the mutation had a higher Ca2+ sensitivity (EC(50)G159D/donor=0.55 +/- 0.13), whereas the maximally activated sliding speed was unaltered. In addition, the cTnC G159D mutation blunted the change in Ca2+ sensitivity when TnI was dephosphorylated. With wild-type troponin, Ca2+ sensitivity was increased (EC50 P/unP=4.7 +/- 1.9) but not with cTnC G159D troponin (EC50 P/unP=1.2 +/- 0.1).Conclusions-We propose that uncoupling of the relationship between phosphorylation and Ca2+ sensitivity could be the cause of the dilated cardiomyopathy phenotype. The differences between these data and previous in vitro results show that native phosphorylation of troponin I and troponin T and other posttranslational modifications of sarcomeric proteins strongly influence the functional effects of a mutation. (Circ Heart Fail. 2009;2:456-464.

Interaction of maternal photoperiod history and food type on growth and reproductive development of laboratory meadow voles (Microtus pennsylvanicus)

The interaction of maternal photoperiod history and four diets were tested by measuring body growth, reproductive development, and pelage development in 9-week-old juvenile meadow voles. Meadow vole dams were housed in long daylengths (LD; 14 h light/day), short daylengths for 2 weeks (SD; 10 h light/day), or short daylengths for 26 weeks (PR; photorefractory) prior to mating. Immediately following parturition, one of four diets was available to dams and pups; (a) a control diet containing no 6-methoxy-2-benzoxazolinone (6-MBOA); (b) the control diet plus sprouted wheat (which contains 6-MBOA); (c) the control diet plus alfalfa harvested in spring (no 6-MBOA); and (d) the control diet plus alfalfa harvested in autumn (no 6-MBOA). By 9 weeks of age, juvenile meadow voles born to photorefractory dams and fed either spring or fall alfalfa or sprouted wheat were significantly larger and more had achieved puberty than juveniles fed only the control diet. Juveniles born to LD dams demonstrated a smaller increase in developmental rate than photorefractory juveniles when fed alfalfa and spring wheat, and juveniles of SD dams showed the smallest effect of alfalfa and sprouted wheat on development. Supplements of spring wheat and both forms of alfalfa had similar positive effects on growth and reproduction. The authors suggest that juvenile meadow voles rely on the interaction of maternal photoperiod history and the availability of nutrient-rich food such as sprouted wheat and alfalfa to time the onset of growth and puberty.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/60640/1/interaction_of_maternal.pd

Circulating Cell-Free DNA Captures the Intratumor Heterogeneity in Multinodular Hepatocellular Carcinoma.

PURPOSE Hepatocellular carcinoma (HCC) is a highly heterogeneous disease, with more than 40% of patients initially diagnosed with multinodular HCCs. Although circulating cell-free DNA (cfDNA) has been shown to effectively detect somatic mutations, little is known about its utility to capture intratumor heterogeneity in patients with multinodular HCC undergoing systemic treatment. MATERIALS AND METHODS Tumor biopsies and plasma were synchronously collected from seven prospectively recruited patients with HCC before and during systemic therapy. Plasma-derived cfDNA and matched germline were subjected to high-depth targeted sequencing with molecular barcoding. The mutational profile of the cfDNA was compared with whole-exome sequencing from matched tumor biopsies. RESULTS Genomic data revealed that out of the seven patients, five were considered intrahepatic metastasis and two multicentric HCCs. cfDNA captured the majority of mutations in the tumors and detected significantly more mutations than tumor biopsies. Driver mutations such as CTNNB1 S33C, NRAS Q61R, ARID1A R727fs, and NF1 E2368fs as well as standard-of-care biomarkers of response to targeted therapy were detected only in cfDNA. In the two patients with multicentric HCC, cfDNA detected mutations derived from the genetically independent and spatially distinct nodules. Moreover, cfDNA was not only able to capture clonal mutations but also the subclonal mutations detected in only one of the multiple biopsied nodules. Furthermore, serial cfDNA detected variants of tumor origin emerging during treatment. CONCLUSION This study revealed that the genetic analysis of cfDNA captures the intratumor heterogeneity in multinodular HCC highlighting the potential for cfDNA as a sensitive and noninvasive tool for precision medicine

Correction to: Tumour suppressor EP300, a modulator of paclitaxel resistance and stemness, is downregulated in metaplastic breast cancer

In the original publication, Fig. 1 depicting the blot for EP300 in CAL51 cells (Fig. 1c) was unintentionally duplicated with that from MDA-MB-231 cells (Fig. 1d). The new figure given in this erratum depicts the correct EP300 blot in Fig. 1c

Populationsgröße, Trichterdichte und Habitatpräferenz der Dünen-Ameisenjungfer Myrmeleon bore (Tjeder, 1941) im Gebiet der Dresdner Heide (Neuroptera) - 2021

Nach dem Erstnachweis von Myrmeleon-Bohrungen (Tjeder, 1941) in der Dresdner Heide im Jahr 2019 (KURTH 2020, Sächs. Entomol. Z. 10: 71-80) wurde die Populationsgröße und Dichte der Art bestimmt. M. bore wurde hauptsächlich in offenen, spärlich bewachsenen, sandigen Gebieten mit direkter Sonneneinstrahlung gefunden. Die flächengewichtete Dichte des gesamten Untersuchungsgebietes (4,05 Hektar) betrug 0,177 Larven/m2. Schätzungen der Populationsgröße basierend auf zufälligen Quadratzahlen führen zu einer Zahl von 4000-7000 Individuen - die größte bekannte Population dieser Art. Die für diese Art aus Laborversuchen bekannte positive Korrelation zwischen Larvengröße und Grubendurchmesser wurde an unserem Studienstandort bestätigt. Diese Korrelation könnte es Forschern ermöglichen, die Altersstruktur von Wildpopulationen abzuschätzen. Angesichts der besonderen Verantwortung Deutschlands für den Schutz dieser Art und der Größe der Population fordern wir den Schutz des Gebietes und eine Priorisierung gegenüber anderen geschützten Arten in diesem Gebiet.Following the first record of Myrmeleon bore (Tjeder, 1941) in the Dresden Heath area in 2019 (KURTH 2020, Sächs. Entomol. Z. 10: 71-80), the population size and density of the species was determined. M. bore mainly was found in open, sparsely vegetated, sandy areas with direct sunlight exposure. The area-weighted density of the entire study site (4.05 hectares) was 0.177 larvae/m2. Population size estimates based on random quadrat counts lead to a figure of 4000-7000 individuals - the largest known population of this species. The positive correlation between larval size and pit diameter known for this species from laboratory trials was confirmed at our study site. This correlation may allow researchers to estimate the age structure of wild populations. Given the special responsibility of Germany for the protection of this species and the size of the population, we urge the protection of the site and a prioritisation over other protected species found in the area

Lexicality and frequency in specific language impairment: accuracy and error data from two nonword repetition tests

Purpose: Deficits in phonological working memory and deficits in phonological processing have both been considered potential explanatory factors in Specific Language Impairment (SLI). Manipulations of the lexicality and phonotactic frequency of nonwords enable contrasting predictions to be derived from these hypotheses. Method: 18 typically developing (TD) children and 18 children with SLI completed an assessment battery that included tests of language ability, non-verbal intelligence, and two nonword repetition tests that varied in lexicality and frequency. Results: Repetition accuracy showed that children with SLI were unimpaired for short and simple high lexicality nonwords, whereas clear impairments were shown for all low lexicality nonwords. For low lexicality nonwords, greater repetition accuracy was seen for nonwords constructed from high over low frequency phoneme sequences. Children with SLI made the same proportion of errors that substituted a nonsense syllable for a lexical item as TD children, and this was stable across nonword length. Conclusions: The data show support for a phonological processing deficit in children with SLI, where long-term lexical and sub-lexical phonological knowledge mediate the interpretation of nonwords. However, the data also suggest that while phonological processing may provide a key explanation of SLI, a full account is likely to be multi-faceted

Caracterização e ocorrência do distúrbio do amolecimento precoce em mamões 'Golden'

The occurrence of green skin and soft pulp in 'Golden' papaya fruit during certain seasons has been reported by farmers in the northern of the state of Espirito Santo, Brazil. The objective of this study was to characterize and determine the occurrence of this disorder, which was referred as "early softening disorder". Fruits were harvested weekly for 11 months (from September to July). The fruits were stored at 10°C, and then fruit flesh firmness and skin color were analyzed. The results of the firmness test were submitted to regression analysis assuming a linear trendline. The slope of the curve was called the 'softening index' (SI). Fruits with early softening are characterized by a loss of firmness in less than 10 days, even when stored under refrigeration. Although softened, the skin of the fruit remains partially green. Fruits with the disorder occurred more frequently from mid-summer to mid-autumn (February to May). It is not possible to distinguish early softening disorder fruits from those without the disorder by skin color and flesh firmness analysis at the time of the harvest.Tem sido relatado por produtores da região norte do Espírito Santo a ocorrência de mamões 'Golden' com casca verde e polpa mole, em determinadas épocas do ano. O objetivo deste trabalho foi caracterizar e determinar a ocorrência deste distúrbio denominado de amolecimento precoce. Foram realizadas coletas semanais durante 11 meses (período de setembro a julho). Os frutos foram armazenados a 10°C e analisados quanto à firmeza da polpa e à cor da casca. Os resultados de firmeza da polpa foram submetidos à análise de regressão, assumindo-se que a equação é do tipo linear, e o ângulo de inclinação da curva foi chamado Índice de Amolecimento (IA). Frutos com o distúrbio caracterizaram-se pela perda da firmeza em menos de 10 dias, mesmo quando armazenados sob refrigeração. Embora amolecidos, a coloração da casca manteve-se parcialmente verde. A maior frequência de frutos com o distúrbio ocorreu de meados de verão a meados de outono (fevereiro a maio). Não é possível distinguir frutos com o distúrbio do amolecimento precoce daqueles normais pela análise da cor da casca e da firmeza da polpa, no momento da colheita

The molecular phenotype of human cardiac myosin associated with hypertrophic obstructive cardiomyopathy

AIM: The aim of the study was to compare the functional and structural properties of the motor protein, myosin, and isolated myocyte contractility in heart muscle excised from hypertrophic cardiomyopathy patients by surgical myectomy with explanted failing heart and non-failing donor heart muscle. METHODS: Myosin was isolated and studied using an in vitro motility assay. The distribution of myosin light chain-1 isoforms was measured by two-dimensional electrophoresis. Myosin light chain-2 phosphorylation was measured by sodium dodecyl sulphate-polyacrylamide gel electrophoresis using Pro-Q Diamond phosphoprotein stain. RESULTS: The fraction of actin filaments moving when powered by myectomy myosin was 21% less than with donor myosin (P = 0.006), whereas the sliding speed was not different (0.310 +/- 0.034 for myectomy myosin vs. 0.305 +/- 0.019 microm/s for donor myosin in six paired experiments). Failing heart myosin showed 18% reduced motility. One myectomy myosin sample produced a consistently higher sliding speed than donor heart myosin and was identified with a disease-causing heavy chain mutation (V606M). In myectomy myosin, the level of atrial light chain-1 relative to ventricular light chain-1 was 20 +/- 5% compared with 11 +/- 5% in donor heart myosin and the level of myosin light chain-2 phosphorylation was decreased by 30-45%. Isolated cardiomyocytes showed reduced contraction amplitude (1.61 +/- 0.25 vs. 3.58 +/- 0.40%) and reduced relaxation rates compared with donor myocytes (TT(50%) = 0.32 +/- 0.09 vs. 0.17 +/- 0.02 s). CONCLUSION: Contractility in myectomy samples resembles the hypocontractile phenotype found in end-stage failing heart muscle irrespective of the primary stimulus, and this phenotype is not a direct effect of the hypertrophy-inducing mutation. The presence of a myosin heavy chain mutation causing hypertrophic cardiomyopathy can be predicted from a simple functional assay

Maximum rates of N2 fixation and primary production are out of phase in a developing cyanobacterial bloom in the Baltic Sea

Although N2-fixing cyanobacteria contribute significantly to oceanic sequestration of atmospheric CO2, little is known about how N2 fixation and carbon fixation (primary production) interact in natural populations of marine cyanobacteria. In a developing cyanobacterial bloom in the Baltic Sea, rates of N2 fixation (acetylene reduction) showed both diurnal and longer-term fluctuations. The latter reflected fluctuations in the nitrogen status of the cyanobacterial population and could be correlated with variations in the ratio of acetylene reduced to 15N2 assimilated. The value of this ratio may provide useful information about the release of newly fixed nitrogen by a cyanobacterial population. However, although the diurnal fluctuations in N2 fixation broadly paralleled diurnal fluctuations in carbon fixation, the longer-term fluctuations in these two processes were out of phase