Variants in the ATM‐CHEK2‐BRCA1 axis determine genetic predisposition and clinical presentation of papillary thyroid carcinoma

The risk of developing papillary thyroid carcinoma (PTC), the most frequent form of thyroid malignancy, is elevated up to 8.6‐fold in first‐degree relatives of PTC patients. The familial risk could be explained by high‐penetrance mutations in yet unidentified genes, or polygenic action of low‐penetrance alleles. Since the DNA‐damaging exposure to ionizing radiation is a known risk factor for thyroid cancer, polymorphisms in DNA repair genes are likely to affect this risk. In a search for low‐penetrance susceptibility alleles we employed Sequenom technology to genotype deleterious polymorphisms in ATM, CHEK2, and BRCA1 in 1,781 PTC patients and 2,081 healthy controls. As a result of the study, we identified CHEK2 rs17879961 (OR = 2.2, P = 2.37e‐10) and BRCA1 rs16941 (odds ratio [OR] = 1.16, P = 0.005) as risk alleles for PTC. The ATM rs1801516 variant modifies the risk associated with the BRCA1 variant by 0.78 (P = 0.02). Both the ATM and BRCA1 variants modify the impact of male gender on clinical variables: T status (P = 0.007), N status (P = 0.05), and stage (P = 0.035). Our findings implicate an important role of variants in the ATM‐ CHEK2‐ BRCA1 axis in modification of the genetic predisposition to PTC and its clinical manifestations. © 2014 Wiley Periodicals, Inc.     

Deep Sequencing Analysis Reveals Temporal Microbiota Changes Associated with Development of Bovine Digital Dermatitis

Bovine digital dermatitis (DD) is a leading cause of lameness in dairy cattle throughout the world. Despite 35 years of research, the definitive etiologic agent associated with the disease process is still unknown. Previous studies have demonstrated that multiple bacterial species are associated with lesions, with spirochetes being the most reliably identified organism. This study details the deep sequencing-based metagenomic evaluation of 48 staged DD biopsy specimens collected during a 3-year longitudinal study of disease progression. Over 175 million sequences were evaluated by utilizing both shotgun and 16S metagenomic techniques. Based on the shotgun sequencing results, there was no evidence of a fungal or DNA viral etiology. The bacterial microbiota of biopsy specimens progresses through a systematic series of changes that correlate with the novel morphological lesion scoring system developed as part of this project. This scoring system was validated, as the microbiota of each stage was statistically significantly different from those of other stages (P < 0.001). The microbiota of control biopsy specimens were the most diverse and became less diverse as lesions developed. Although Treponema spp. predominated in the advanced lesions, they were in relatively low abundance in the newly described early lesions that are associated with the initiation of the disease process. The consortium of Treponema spp. identified at the onset of disease changes considerably as the lesions progress through the morphological stages identified. The results of this study support the hypothesis that DD is a polybacterial disease process and provide unique insights into the temporal changes in bacterial populations throughout lesion development.     

Influence of ezetimibe on selected parameters of oxidative stress in rat liver subjected to ischemia/reperfusion

Introduction

Ischemia/reperfusion (I/R) is considered to be one of the main causes of liver damage after transplantation. The authors evaluated the effect of ezetimibe on selected oxidative stress parameters in ischemic/reperfused (I/R) rat liver.

Material and methods

Rats were administered ezetimibe (5 mg/kg) (groups E and E-I/R) or saline solution (groups C and C-I/R) intragastrically for 21 days. Livers of animals in groups C-I/R and E-I/R were subjected to 60 min of partial ischemia (left lateral and median lobes) followed by 4 h of reperfusion. Alanine and asparagine aminotransferase (ALT, AST) activity was determined in blood before I/R and during reperfusion (at 15 and 240 min). After the reperfusion period, malondialdehyde (MDA), superoxide dismutase (SOD), glutathione (GSH) and glutathione peroxidase (GPx) were determined in liver homogenates using colorimetric methods.

Results

Ezetimibe caused a significant increase in GSH level in groups subjected to I/R (E-I/R (99.91 ±9.01) vs. C-I/R (90.51 ±8.87), p < 0.05). Additionally, under I/R the decrease of GPx activity in the drug-treated group was lower compared to the non-treated group (E-I/R (3.88 ±1.11) vs. E (5.31 ±1.83), p = 0.076). Neither ezetimibe nor I/R affected SOD or MDA levels. I/R produced a significant increase in aminotransferase levels (ALT240-0: C-I/R (42.23 ±43.56) vs. C (9.75 ±11.09), and E-I/R (39.85 ±26.53) vs. E (4.38 ±1.36), p < 0.05 in both cases; AST 240-0: E-I/R (53.87 ±17.23) vs. E (24.10 ±9.66), p < 0.05) but no effect of ezetimibe on those enzymes was found.

Conclusions

Ezetimibe demonstrates antioxidant properties in rat livers subjected to I/R. However, neither a hepatoprotective nor a hepatotoxic effect of ezetimibe was demonstrated, regardless of I/R.     

Crystal structure of a Fanconi anemia-associated nuclease homolog bound to 5′ flap DNA: basis of interstrand cross-link repair by FAN1

Fanconi anemia (FA) is an autosomal recessive genetic disorder caused by defects in any of 15 FA genes responsible for processing DNA interstrand cross-links (ICLs). The ultimate outcome of the FA pathway is resolution of cross-links, which requires structure-selective nucleases. FA-associated nuclease 1 (FAN1) is believed to be recruited to lesions by a monoubiquitinated FANCI–FANCD2 (ID) complex and participates in ICL repair. Here, we determined the crystal structure of Pseudomonas aeruginosa FAN1 (PaFAN1) lacking the UBZ (ubiquitin-binding zinc) domain in complex with 5′ flap DNA. All four domains of the right-hand-shaped PaFAN1 are involved in DNA recognition, with each domain playing a specific role in bending DNA at the nick. The six-helix bundle that binds the junction connects to the catalytic viral replication and repair (VRR) nuclease (VRR nuc) domain, enabling FAN1 to incise the scissile phosphate a few bases distant from the junction. The six-helix bundle also inhibits the cleavage of intact Holliday junctions. PaFAN1 shares several conserved features with other flap structure-selective nucleases despite structural differences. A clamping motion of the domains around the wedge helix, which acts as a pivot, facilitates nucleolytic cleavage. The PaFAN1 structure provides insights into how archaeal Holliday junction resolvases evolved to incise 5′ flap substrates and how FAN1 integrates with the FA complex to participate in ICL repair.     

Exercise volume and intensity: a dose–response relationship with health benefits

Introduction

The health benefits of exercise are well established. However, the relationship between exercise volume and intensity and health benefits remains unclear, particularly the benefits of low-volume and intensity exercise.

Purpose

The primary purpose of this investigation was, therefore, to examine the dose–response relationship between exercise volume and intensity with derived health benefits including volumes and intensity of activity well below international recommendations.

Methods

Generally healthy, active participants (n = 72; age = 44 ± 13 years) were assigned randomly to control (n = 10) or one of five 13-week exercise programs: (1) 10-min brisk walking 1×/week (n = 10), (2) 10-min brisk walking 3×/week (n = 10), (3) 30-min brisk walking 3×/week (n = 18), (4) 60-min brisk walking 3×/week (n = 10), and (5) 30-min running 3×/week (n = 14), in addition to their regular physical activity. Health measures evaluated pre- and post-training including blood pressure, body composition, fasting lipids and glucose, and maximal aerobic power (VO₂max).

Results

Health improvements were observed among programs at least 30 min in duration, including body composition and VO₂max: 30-min walking 28.8–34.5 mL kg^?1 min^?1, 60-min walking 25.1–28.9 mL kg^?1 min^?1, and 30-min running 32.4–36.4 mL kg^?1 min^?1. The greater intensity running program also demonstrated improvements in triglycerides.

Conclusion

In healthy active individuals, a physical activity program of at least 30 min in duration for three sessions/per week is associated with consistent improvements in health status.     

Thymoma with loss of keratin expression (and giant cells): a potential diagnostic pitfall

Due to its profound therapeutic consequences, the distinction between thymoma and T-lymphoblastic lymphoma in needle biopsies is one of the most challenging in mediastinal pathology. One essential diagnostic criterion favouring thymoma is the demonstration of increased numbers of keratin-positive epithelial cells by immunohistochemistry. Loss of keratin expression in neoplastic epithelial cells could lead to detrimental misdiagnoses. We here describe a series of 14 thymic epithelial tumours (11 type B2 and B3 thymomas, 3 thymic carcinomas) with loss of expression of one or more keratins. Cases were analysed for expression of various keratins and desmosomal proteins by immunohistochemistry and immunofluorescence and compared with 45 unselected type B thymomas and 24 thymic carcinomas arranged in a multitissue histological array. All 14 cases showed highly reduced expression of at least one keratin, three cases were completely negative for all keratins studied. Of the 14 cases, 13 showed strong nuclear expression of p63. Expression of desmosomal proteins was preserved, suggesting intact cell contact structures. Loss of expression of broad-spectrum-keratins and K19 was observed in 3 and 5 % of unselected thymomas and in 30 and 60 % of thymic carcinomas. A proportion of keratin-depleted thymomas contained giant cells, reminiscent of thymic nurse cells. Loss of keratin expression in type B2 and B3 thymomas is an important diagnostic pitfall in the differential diagnosis with T-lymphoblastic lymphoma and can be expected in 5 % of cases. A panel of epithelial markers including p63 is warranted to ensure correct diagnosis of keratin-negative mediastinal tumours.     

Parietal Cell Carcinoma of Gastric Cardia: Immunophenotype and Ultrastructure

The case is reported of a clinically aggressive parietal cell carcinoma of the gastric cardia in a 67-year-old man. Histologically, the tumor was a poorly differentiated adenocarcinoma with a predominantly solid growth pattern, though with areas exhibiting glandular morphology and with extensive lymphatic invasion. The tumor cells had eosinophilic, finely granular cytoplasm, with focal Alcian blue-positive mucin in the gland lumens. Ultrastructural examination of the pleural metastasis and gastrectomy specimen demonstrated many mitochondria, tubulovesicular profiles of endoplasmic reticulum, and intracytoplasmic lumens, which resembled intracellular canaliculi of parietal cells. Immunohistochemically, there was positive staining of tumor cells for the parietal cell specific antibodies to H/K-ATPase and human milk fat globule-2 (HMFG-2).     

The influence of dance training on growth and maturation of young females: A mixed longitudinal study

Background: The effects of systematic sports training during childhood and adolescence on subsequent growth and sexual maturation remains in dispute.

Aim: The study aimed to determine whether moderate–high volumes of dance training adversely influence linear growth and sexual maturation of young girls progressing through puberty.

Subjects and methods: This 3-year mixed longitudinal study comprised 82 novice dancers and 61 controls, aged 8–11 years at baseline, who were assessed bi-annually for 3 consecutive years. A biological maturational age was determined by estimating attainment of age at peak height velocity (PHV). Body dimensions were measured by anthropometry, and exercise levels, nutritional intake and age at menarche by questionnaires.

Results: Controls had significantly greater unadjusted height velocity than dancers 1 year before PHV, however there was no difference between groups in age of attainment of PHV. When controlling for maturation, lean mass, fat mass and extracurricular sport (excluding dancing), there were no group differences in absolute growth or velocity of growth in height, sitting height or leg length. Within the dancers there were no effects of years of dancing (>6.5 years) or weekly dance hours (>7?h per week) on growth velocities. No association was found between age at menarche and years or hours of dance training.

Conclusion: Results suggest that moderate–high levels of dance training do not affect linear growth and maturation. Thus, girls should not be discouraged from dance participation on the basis of potential growth delays.

Résumé. Arrière plan: On reste partagé sur les effets que l’entraînement sportif systématique pendant l’enfance et l’adolescence, peut avoir sur la croissance et sur la maturation sexuelle.

Objectif: L’étude cherche à savoir si l’entraînement moyen ou intensif à la danse, influence négativement la croissance linéaire et la maturation sexuelle des filles en cours de puberté.

Sujets et méthodes: Cette étude semi longitunale de trois ans impliquait 82 danseuses novices et 61 contrôles, âgées de 8 à 11 ans au départ, qui ont été examinées deux fois par an pendant trois années consécutives. Un âge de maturation biologique a été déterminé en estimant à quel âge survenait la croissance staturale de pic (CSP). Les dimensions corporelles ont été mesurées par anthropométrie et les niveaux d’activité physique, ainsi que l’alimentation et l’âge aux premières règles, ont été renseignés par questionnaires.

Résultats: Les contrôles présentent une croissance staturale non ajustée significativement plus élevée que celle des danseuses, un an avant la CSP, mais il n’y a cependant pas de différence d’âge d’atteinte de la CSP entre les groupes. A maturation, masse maigre, masse grasse et sport non programmé constants (à l’exclusion de la danse), il n’y a pas de différences entre les groupes, ni en croissance absolue, ni en vélocité de croissance staturale, taille assis ou longueur de la jambe. Parmi les danseuses, on n’observe aucun effet du nombre d’années de danse (>6,5 ans) ou du nombre d’heures de danse hebdomadaire (>7h par semaine) sur les vitesses de croissance. On n’a pas trouvé d’association entre le nombre d’heures d’entraînement à la danse et l’âge aux premières règles.

Conclusion: Les résultats suggèrent que des niveaux d’entraînement à la danse de modérés à élevés n’affectent pas la croissance linéaire ni la maturation. Les filles ne doivent donc pas être dissuadées de pratiquer la danse sur la crainte d’un retard de croissance potentiel.

Zusammenfassung. Hintergrund: Die Wirkung systematischen sportlichen Trainings während Kindheit und Adoleszenz auf das folgende Wachstum und die sexuelle Reifung wird noch kontrovers diskutiert.

Ziel: Das Ziel der Untersuchung war die Bestimmung, ob mäßig bis sehr intensives Tanztraining einen nachteiligen Einfluss auf das lineare Wachstum und die sexuelle Reifung junger Mädchen während ihrer pubertären Entwicklung hat.

Probanden und Methoden: Diese dreijährige, gemischt longitudinale Studie umfasste 82 neu anfangende Tänzerinnen und 61 Kontrollen, die zu Beginn der Studie 8 bis 11 Jahre alt waren und hernach in 6-montigen Abständen für 3 weitere Jahre untersucht wurden. Ein biologisches Reifealter wurde durch Schätzung des Alters zum Zeitpunkt der maximalen Wachstumsgeschwindigkeit (peak height velocity, PHV) bestimmt. Körpermaße wurden mittels Anthropometrie bestimmt, sowie sportliche Leistung, Ernährung und Menarchealter mittels Fragebogen.

Ergebnisse: Unkorrigiert hatten Kontrollen eine signifikant größere Wachstumsgeschwindigkeit als Tänzerinnen ein Jahr vor PHV, allerdings gab es keinen Unterschied zwischen den Gruppen hinsichtlich des Alters bei Erreichen der PHV. Wenn für Reifestatus, fettfreie Körpermasse, Fettmasse und zusätzliche sportliche Aktivität (neben dem Tanzen) korrigiert wurde, gab es keine Gruppenunterschiede mehr hinsichtlich der absoluten Zunahme oder der Wachstumsgeschwindigkeit von Körperhöhe, Sitzhöhe oder Beinlänge. Unter den Tänzerinnen gab es keine Unterschiede hinsichtlich der Anzahl der Tanzjahre (>6,5?Jahre) oder der wöchentlichen Tanzstunden (>7?h?pro Woche) auf die Wachstumsgeschwindigkeit. Es wurden keine Beziehungen zwischen Menarchealter und der Anzahl der Tanzjahre oder der wöchentlichen Tanztrainingsstunden gefunden.

Zusammenfassung: Die Ergebnisse sprechen dafür, dass ein mäßig bis sehr intensives Tanztraining keinen Einfluss auf lineares Wachstum und körperliche Reifung hat. Aus diesem Grunde sollten Mädchen nicht von der Teilnahme am Tanzen abgeschreckt werden, nur weil eine mögliche Beeinträchtigung des Wachstums befürchtet wird.

Resumen. Antecedentes: Los efectos del entrenamiento deportivo sistemático durante la infancia y adolescencia sobre el crecimiento y la maduración sexual siguen siendo objeto de debate.

Objetivo: El estudio intenta determinar si los niveles moderados y/o altos de entrenamiento en danza influyen adversamente sobre el crecimiento lineal y la maduración sexual de las chicas jóvenes que están progresando hacia la pubertad.

Sujetos y métodos: Este estudio semi-longitudinal de 3 años de duración incluía a 82 bailarinas principiantes y 61 controles, de entre 8 y 11 años de edad como punto de comparación, que fueron estudiadas bianualmente durante tres años consecutivos. La edad biológica de maduración se determinó estimando la edad a la que se alcanzaba el pico de velocidad en el crecimiento estatural (PHV). Las dimensiones corporales fueron medidas por antropometría, y los niveles de ejercicio, la ingesta nutricional y la edad de menarquia mediante cuestionarios.

Resultados: Las chicas control tenían velocidades estaturales no ajustadas significativamente mayores que las bailarinas 1 año antes del PHV, aunque no había diferencias significativas entre los grupos en cuanto a la edad a la que se alcanzaba el PHV. Cuando se controlaba para la maduración, la masa magra, la masa grasa y el deporte extra-curricular (excluyendo la danza), no había diferencias entre los grupos en el crecimiento absoluto o en la velocidad de crecimiento en estatura, talla sentado o longitud de la pierna. Entre las bailarinas, no había efectos debidos a los años que llevaban bailando (> 6,5 años) o las horas de baile semanales (> 7 horas/semana ) sobre las velocidades de crecimiento. No se encontró ninguna asociación entre la edad de menarquia y los años u horas de entrenamiento en danza.

Conclusión: Los resultados sugieren que los niveles moderados y/o altos de entrenamiento en danza no afectan ni al crecimiento lineal ni a la maduración. Por tanto, las chicas no deberían renunciar a practicar danza basándose en supuestos retrasos de su potencial de crecimiento.     

Optimization of the Cardiovascular Therapeutic Properties of Mesenchymal Stromal/Stem Cells–Taking the Next Step

Despite current treatment options, cardiac failure is associated with significant morbidity and mortality highlighting a compelling clinical need for novel therapeutic approaches. Based on promising pre-clinical data, stem cell therapy has been suggested as a possible therapeutic strategy. Of the candidate cell types evaluated, mesenchymal stromal/stem cells (MSCs) have been widely evaluated due to their ease of isolation and ex vivo expansion, potential allogeneic utility and capacity to promote neo-angiogenesis and endogenous cardiac repair. However, the clinical application of MSCs for mainstream cardiovascular use is currently hindered by several important limitations, including suboptimal retention and engraftment and restricted capacity for bona fide cardiomyocyte regeneration. Consequently, this has prompted intense efforts to advance the therapeutic properties of MSCs for cardiovascular disease. In this review, we consider the scope of benefit from traditional plastic adherence-isolated MSCs and the lessons learned from their conventional use in preclinical and clinical studies. Focus is then given to the evolving strategies aimed at optimizing MSC therapy, including discussion of cell-targeted techniques that encompass the preparation, pre-conditioning and manipulation of these cells ex vivo, methods to improve their delivery to the heart and innovative substrate-directed strategies to support their interaction with the host myocardium.     

Exercise-induced endocannabinoid signaling is modulated by intensity

Endocannabinoids (eCB) are endogenous ligands for cannabinoid receptors that are densely expressed in brain networks responsible for reward. Recent work shows that exercise activates the eCB system in humans and other mammals, suggesting eCBs are partly responsible for the reported improvements in mood and affect following aerobic exercise in humans. However, exercise-induced psychological changes reported by runners are known to be dependent on exercise intensity, suggesting that any underlying molecular mechanism should also change with varying levels of exercise intensity. Here, we examine circulating levels of eCBs following aerobic exercise (treadmill running) in recreationally fit human runners at four different intensities. We show that eCB signaling is indeed intensity dependent, with significant changes in circulating eCBs observed following moderate intensities only (very high and very low intensity exercises do not significantly alter circulating eCB levels). Our results are consistent with intensity-dependent psychological state changes with exercise and therefore support the hypothesis that eCB activity is related to neurobiological effects of exercise. Thus, future studies examining the role of exercise-induced eCB signaling on neurobiology or physiology must take exercise intensity into account.