Assembling DNA Plasmids with the Multi-Kingdom (MK) Cloning System.

The Golden Gate cloning technique is used to assemble DNA parts into higher-order assemblies. Individual parts containing compatible overhangs generated by type IIS restriction enzymes are joined together using DNA ligase. The technique enables users to assemble custom transcription units (TUs) for a wide array of experimental assays.

Arabidopsis CML13 and CML14 Have Essential and Overlapping Roles in Plant Development.

Calmodulin (CaM)-like proteins (CMLs) are the largest family of calcium-binding proteins in plants, yet the functions of most CMLs are unknown. Arabidopsis CML13 and CML14 are closely related paralogs that interact with the isoleucine-glutamine (IQ) domains of myosins, IQ-domain proteins and CaM-binding transcription activators (CAMTAs). Here, we explored the physiological roles of CML13 and CML14 during development by using dexamethasone (Dex)-inducible RNA silencing to suppress either CML13 or CML14 transcript levels.

Arbuscular-Mycorrhizal Symbiosis in Regulated by the Transcription Factor MtbHLHm1;1 and the Ammonium Facilitator Protein MtAMF1;3.

Root systems of most land plants are colonised by arbuscular mycorrhiza fungi. The symbiosis supports nutrient acquisition strategies predominantly associated with plant access to inorganic phosphate. The nutrient acquisition is enhanced through an extensive network of external fungal hyphae that extends out into the soil, together with the development of fungal structures forming specialised interfaces with root cortical cells.

Decreased left heart flow in fetal lambs causes left heart hypoplasia and pro-fibrotic tissue remodeling.

Low blood flow through the fetal left heart is often conjectured as an etiology for hypoplastic left heart syndrome (HLHS). To investigate if a decrease in left heart flow results in growth failure, we generate left ventricular inflow obstruction (LVIO) in mid-gestation fetal lambs by implanting coils in their left atrium using an ultrasound-guided percutaneous technique. Significant LVIO recapitulates important clinical features of HLHS: decreased antegrade aortic valve flow, compensatory retrograde perfusion of the brain and ascending aorta (AAo) from the arterial duct, severe left heart hypoplasia, a non-apex forming LV, and a thickened endocardial layer.

RPG interacts with E3-ligase CERBERUS to mediate rhizobial infection in Lotus japonicus.

Symbiotic interactions between rhizobia and legumes result in the formation of root nodules, which fix nitrogen that can be used for plant growth. Rhizobia usually invade legume roots through a plant-made tunnel-like structure called an infection thread (IT). RPG (Rhizobium-directed polar growth) encodes a coiled-coil protein that has been identified in Medicago truncatula as required for root nodule infection, but the function of RPG remains poorly understood.

Fatal Pediatric Streptococcal Infection: A Clinico-Pathological Study.

Purpose And Context: Streptococcal Infection (SI) is an important cause of pediatric death in children, yet limited reports exist on autopsy findings in fatal SI cases.

Method: Case records (1997-2019) of SI with no pre-existing risk factors were reviewed and selected. Their clinical and pathological findings in the autopsy reports were analyzed.

Serious and Fatal Complications after Neonatal Circumcision.

Neonatal circumcision (NNC) is the most frequently performed surgical procedure worldwide and is generally considered safe in Western societies. Deaths attributed to NNC are seldom reported and are mostly explained by lack of adherence to medical standards. We reviewed our emergency department database for circumcision-related emergency admissions.

Paediatric dilated cardiomyopathy with and without endocardial fibroelastosis - a pathological analysis of 89 explants.

Heart failure due to dilated cardiomyopathy is a major indication for paediatric cardiac transplantation. Endocardial fibroelastosis is a recognised pathological finding of unknown prognostic significance in paediatric dilated cardiomyopathy. To evaluate the nature of the association between left ventricular endocardial fibroelastosis and paediatric dilated cardiomyopathy, we reviewed surgical pathology reports of dilated cardiomyopathy explants (1986-2016) in order to characterise the pathological findings and to compare and contrast their frequency among four age groups: less than 1 year; 1-5 years; 6-10 years; and greater than 11 years.

Lossless Compression of Human Movement IMU Signals.

Real-time human movement inertial measurement unit (IMU) signals are central to many emerging medical and technological applications, yet few techniques have been proposed to process and represent this information modality in an efficient manner. In this paper, we explore methods for the lossless compression of human movement IMU data and compute compression ratios as compared with traditional representation formats on a public corpus of human movement IMU signals for walking, running, sitting, standing, and biking human movement activities. Delta coding was the highest performing compression method which compressed walking, running, and biking data by a factor of 10 and compressed sitting and standing data by a factor of 18 relative to the original CSV formats.

Genetic Diagnosis and the Severity of Cardiovascular Phenotype in Patients With Elastin Arteriopathy.

Background: Elastin insufficiency causes recurrent vascular stenoses. Hemizygous deletion of the elastin gene () causes Williams-Beuren syndrome (WBS), while single nucleotide variants in cause nonsyndromic supravalvar aortic stenosis (SVAS). Our objective was to compare cardiovascular disease outcomes in patients with WBS and nonsyndromic SVAS.

Feasibility of Wearable Haptic Biofeedback Training for Reducing the Knee Abduction Moment During Overground Walking.

Gait modifications are effective in reducing the first peak knee abduction moment (PKAM), a surrogate for knee loading. Reliance on 3D motion capture currently restricts these modifications to the laboratory. Therefore, our purpose was to test the feasibility of a novel wearable biofeedback system to train (1) toe-in and trunk lean modifications and (2) combined toe-in and trunk lean modifications to reduce PKAM during overground walking outside of the laboratory.

Mixed-reality view of cardiac specimens: a new approach to understanding complex intracardiac congenital lesions.

Digital reality is an emerging platform for three-dimensional representation of medical imaging data. In this technical innovation paper, the authors evaluated the accuracy and utility of mixed-reality technology in the morphological evaluation of complex congenital heart disease. The authors converted CT datasets of 12 heart specimens with different subtypes of double-outlet right ventricle to stereoscopic images and interrogated them using a mixed-reality system.

Flow-targeted pediatric ex vivo heart perfusion in donation after circulatory death: A porcine model.

Background: The optimal blood flow and pressure to perfuse pediatric hearts from donation after circulatory death (DCD) on the ex vivo perfusion system has not been elucidated. This study sought to investigate the optimal perfusion strategy for pediatric DCD hearts by using a juvenile porcine model comparing pressure- vs flow-targeted strategy.

Methods: The hearts of the juvenile DCD pigs were explanted, and the coronary arteries were perfused for 2 hours by the ex vivo heart perfusion system with 2 different perfusion strategies; pressure-targeted perfusion (target coronary perfusion pressure: 40 mm Hg, group A) and flow-targeted perfusion (target coronary perfusion flow: 10 ml/kg/min, group B).

Influence of IMU position and orientation placement errors on ground reaction force estimation.

Unlabelled: Wearable inertial measurement units (IMU) have been proposed to estimate GRF outside of specialized laboratories, however the precise influence of sensor placement error on accuracy is unknown. We investigated the influence of IMU position and orientation placement errors on GRF estimation accuracy.

Methods: Kinematic data from twelve healthy subjects based on marker trajectories were used to simulate 1848 combinations of sensor position placement errors (range ± 100 mm) and orientation placement errors (range ± 25°) across eight body segments (trunk, pelvis, left/right thighs, left/right shanks, and left/right feet) during normal walking trials for baseline cases when a single sensor was misplaced and for the extreme cases when all sensors were simultaneously misplaced.

Rebuilding core abscisic acid signaling pathways of Arabidopsis in yeast.

The phytohormone abscisic acid (ABA) regulates plant responses to abiotic stress, such as drought and high osmotic conditions. The multitude of functionally redundant components involved in ABA signaling poses a major challenge for elucidating individual contributions to the response selectivity and sensitivity of the pathway. Here, we reconstructed single ABA signaling pathways in yeast for combinatorial analysis of ABA receptors and coreceptors, downstream-acting SnRK2 protein kinases, and transcription factors.

A unified multi-kingdom Golden Gate cloning platform.

Assembling composite DNA modules from custom DNA parts has become routine due to recent technological breakthroughs such as Golden Gate modular cloning. Using Golden Gate, one can efficiently assemble custom transcription units and piece units together to generate higher-order assemblies. Although Golden Gate cloning systems have been developed to assemble DNA plasmids required for experimental work in model species, they are not typically applicable to organisms from other kingdoms.

Ankyrin-B dysfunction predisposes to arrhythmogenic cardiomyopathy and is amenable to therapy.

Arrhythmogenic cardiomyopathy (ACM) is an inherited arrhythmia syndrome characterized by severe structural and electrical cardiac phenotypes, including myocardial fibrofatty replacement and sudden cardiac death. Clinical management of ACM is largely palliative, owing to an absence of therapies that target its underlying pathophysiology, which stems partially from our limited insight into the condition. Following identification of deceased ACM probands possessing ANK2 rare variants and evidence of ankyrin-B loss of function on cardiac tissue analysis, an ANK2 mouse model was found to develop dramatic structural abnormalities reflective of human ACM, including biventricular dilation, reduced ejection fraction, cardiac fibrosis, and premature death.

Histopathologic Analysis in Sudden Infant and Child Deaths: A Practical Approach.

The forensic pathologist responsible for sudden unexpected death (SUD) investigation in the pediatric setting faces many challenges. It usually takes many years to obtain reasonable experience and exposure to the wide variety of diseases that may present as SUD in a pediatric context, and to appreciate the differences in the etiology and clinical context between the pediatric and adult SUD setting. In pediatric SUD, it is necessary to conduct a systematic, pediatric-focused autopsy investigation including extensive histopathological assessment and ancillary testing.

Alveolar capillary dysplasia with misalignment of the pulmonary veins and hypoplastic left heart sequence caused by an in frame deletion within FOXF1.

Alveolar capillary dysplasia with misalignment of the pulmonary veins (ACDMPV) is a rare, autosomal dominant disorder of interstitial lung development, leading to pulmonary hypertension, and death in infancy. Associated features include malformations of the heart, gastrointestinal tract, and genitourinary system. ACDMPV is caused by heterozygous variants in the FOXF1 gene or microdeletions involving FOXF1.

Influence of skill level on predicting the success of one's own basketball free throws.

Basketball players sometimes claim to know when their shot is good, even before it goes in. This is likely because shooter proprioception can help determine shot outcome, even before their eyes confirm it. This phenomenon, however, has not been systematically explored for collegiate and recreational shooters.

Investigations After Death in Children.

Objectives: To discuss the role of investigations after death in children as part of a supplement on "Death and Dying in the PICU."

Data Sources: Literature review, personal experience, and expert opinion.

Data Selection: Not applicable.

Sudden Unexpected Death in Neonates: A Clinico-pathological Study.

Clinico-pathological studies that focus on sudden unexpected death (SUD) in the neonatal period are rare. The objective of this study was to elucidate the frequency and pathological spectrum of anatomical causes of death (CODs), found in the setting of sudden unexpected death in neonates (SUD-N), and to correlate the COD with premortem circumstantial information. We conducted a detailed review of all autopsy reports on SUD-N cases at our institution from 1997 to 2015.

Attentional dysfunction and recovery in concussion: effects on the P300m and contingent magnetic variation.

Primary Objective: To examine the effect of concussion on indices of attention using magnetoencephalography.

Methods And Procedures: Thirteen patients were recruited from the emergency department and scanned within 3-6 days of injury. Five returned for follow-up scans one and three months post-injury.

A quantitative hypermorphic allele confers ectopic calcium flux and impairs cellular development.

The coordinated control of Ca signaling is essential for development in eukaryotes. Cyclic nucleotide-gated channel (CNGC) family members mediate Ca influx from cellular stores in plants (Charpentier et al., 2016; Gao et al.