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1.
PLoS Genet ; 19(2): e1010621, 2023 02.
Article in English | MEDLINE | ID: mdl-36735729

ABSTRACT

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. In this study, we identified and characterized RPG in Lotus japonicus and determined that it is required for IT formation. RPG was induced by Mesorhizobium loti or purified Nodulation factor and displayed an infection-specific expression pattern. Nodule inception (NIN) bound to the RPG promoter and induced its expression. We showed that RPG displayed punctate subcellular localization in L. japonicus root protoplasts and in root hairs infected by M. loti. The N-terminal predicted C2 lipid-binding domain of RPG was not required for this subcellular localization or for function. CERBERUS, a U-box E3 ligase which is also required for rhizobial infection, was found to be localized similarly in puncta. RPG co-localized and directly interacted with CERBERUS in the early endosome (TGN/EE) compartment and near the nuclei in root hairs after rhizobial inoculation. Our study sheds light on an RPG-CERBERUS protein complex that is involved in an exocytotic pathway mediating IT elongation.


Subject(s)
Lotus , Rhizobium , Rhizobium/genetics , Lotus/genetics , Lotus/metabolism , Ubiquitin-Protein Ligases/genetics , Ubiquitin-Protein Ligases/metabolism , Plant Proteins/genetics , Plant Proteins/metabolism , Symbiosis/genetics , Gene Expression Regulation, Plant , Root Nodules, Plant/genetics , Plant Roots
2.
Plant Cell Physiol ; 65(2): 228-242, 2024 Feb 15.
Article in English | MEDLINE | ID: mdl-37946525

ABSTRACT

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. In the absence of inducible suppression, CML13- and CML14-RNA-interference lines were indistinguishable from wild-type (WT) plants throughout development. In contrast, induction of silencing treatment led to rapid increases in RNA-hairpin production that correlated with a targeted reduction in CML13 or CML14 transcript levels and a range of developmental and morphological effects. RNA-suppression treatment did not impair the germination of CML13- or 14-RNA-interference lines, but these seedlings were chlorotic, displayed high mortality and failed to achieve seedling establishment. Under Dex treatment, seeds of CML13- and CML14-RNA-interference lines exhibited differential sensitivity to exogenous ABA compared to WT seeds. Induced RNA suppression of mature plants led to reduced silique length, shorter roots and rapid leaf senescence in CML13- and 14-RNA-interference plants, which correlated with increased gene expression of the senescence marker Senescence-Associated Gene13 (SAG13). Plants induced for RNA suppression at 2 weeks post-germination exhibited a much stronger phenotype than treatment of 3-, 4- or 5-week-old plants. Collectively, our data indicate that both CML13 and CML14 are essential for normal development and function across a broad range of tissues and developmental stages.


Subject(s)
Arabidopsis Proteins , Arabidopsis , Arabidopsis/metabolism , Arabidopsis Proteins/metabolism , Abscisic Acid/metabolism , Germination/genetics , Seedlings/metabolism , Seeds , RNA/metabolism , Gene Expression Regulation, Plant , Plants, Genetically Modified/genetics
3.
EMBO J ; 38(17): e101859, 2019 09 02.
Article in English | MEDLINE | ID: mdl-31368592

ABSTRACT

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. The analysis shows that some ABA receptors stimulate the pathway even in the absence of ABA and that SnRK2s are major determinants of ABA responsiveness by differing in the ligand-dependent control. Five SnRK2s, including SnRK2.4 known to be active under osmotic stress in plants, activated ABA-responsive transcription factors and were regulated by ABA receptor complexes in yeast. In the plant tissue, SnRK2.4 and ABA receptors competed for coreceptor interaction in an ABA-dependent manner consistent with a tight integration of SnRK2.4 into the ABA signaling pathway. The study establishes the suitability of the yeast system for the dissection of core signaling cascades and opens up future avenues of research on ligand-receptor regulation.


Subject(s)
Abscisic Acid/metabolism , Arabidopsis Proteins/genetics , Arabidopsis/metabolism , Yeasts/growth & development , Arabidopsis/genetics , Arabidopsis Proteins/metabolism , Biosynthetic Pathways , Gene Expression Regulation, Plant , Osmotic Pressure , Phosphorylation , Protein Engineering , Protein Serine-Threonine Kinases/genetics , Yeasts/genetics
4.
Int J Mol Sci ; 24(18)2023 Sep 19.
Article in English | MEDLINE | ID: mdl-37762569

ABSTRACT

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. Orthologs of the bHLHm1;1 transcription factor, previously described in soybean nodules (GmbHLHm1) and linked to the ammonium facilitator protein GmAMF1;3, have been identified in Medicago (Medicago truncatula) roots colonised by AM fungi. Expression studies indicate that transcripts of both genes are also present in arbuscular containing root cortical cells and that the MtbHLHm1;1 shows affinity to the promoter of MtAMF1;3. Both genes are induced by AM colonisation. Loss of Mtbhlhm1;1 expression disrupts AM arbuscule abundance and the expression of the ammonium transporter MtAMF1;3. Disruption of Mtamf1;3 expression reduces both AM colonisation and arbuscule development. The respective activities of MtbHLHm1;1 and MtAMF1;3 highlight the conservation of putative ammonium regulators supporting both the rhizobial and AM fungal symbiosis in legumes.


Subject(s)
Medicago truncatula , Transcription Factors , Transcription Factors/genetics , Symbiosis/genetics , Gene Expression Regulation , Medicago truncatula/genetics , Nutrients
5.
Pediatr Dev Pathol ; 25(4): 409-418, 2022.
Article in English | MEDLINE | ID: mdl-35227107

ABSTRACT

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. RESULTS: In our cohort of 38 cases based on bacterial culture results, SI was most commonly caused by Streptococcus pneumoniae (SPn; 45%) and Streptococcus pyogenes (SPy; 37%). 92% of decedents had some prodromal symptoms prior to terminal presentation. The clinical course was often rapid, with 89% found unresponsive, suddenly collapsing, or dying within 24 hours of hospital admission. 64% of deaths were attributed to sepsis, more frequently diagnosed in the SPy group than in the SPn group (71% vs 48%). Pneumonia was found in both SPn and SPy groups, whereas meningitis was exclusively associated with SPn. CONCLUSION: Our study shows fatal SI is most commonly caused by either SPn or SPy, both of which are frequently associated with prodromal symptoms, rapid terminal clinical course, and evidence of sepsis. Postmortem diagnosis of sepsis is challenging and should be correlated with clinical features, bacterial culture results, and autopsy findings.


Subject(s)
Streptococcal Infections , Autopsy , Cause of Death , Child , Humans , Prodromal Symptoms , Sepsis/diagnosis , Sepsis/microbiology , Sepsis/mortality , Streptococcal Infections/diagnosis , Streptococcal Infections/microbiology , Streptococcal Infections/mortality , Streptococcus pneumoniae/isolation & purification , Streptococcus pyogenes/isolation & purification
6.
Cardiol Young ; 32(7): 1041-1047, 2022 Jul.
Article in English | MEDLINE | ID: mdl-34486505

ABSTRACT

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. The 89 explants (47 males and 42 females) were all characterised by increased weight and left ventricular chamber dilatation without increased wall thickness. Ninety-five per cent of the specimens in the two youngest subsets had left ventricular endocardial fibroelastosis. Compared to the oldest age group, recipients aged 1-5 years had a 6-fold increase and those younger than 1 year a 19-fold increase in the odds of observing left ventricular endocardial fibroelastosis. Explants with and without endocardial fibroelastosis were otherwise phenotypically similar. In paediatric dilated cardiomyopathy endocardial fibroelastosis is a very common pathological finding, especially in infants and young children. We propose that the descriptive, clinico-pathological designation "Dilated Cardiomyopathy with Endocardial Fibroelastosis" should be adopted to facilitate future investigation into the potential prognostic/therapeutic significance of left ventricular endocardial fibroelastosis.


Subject(s)
Cardiomyopathy, Dilated , Endocardial Fibroelastosis , Heart Transplantation , Cardiomegaly , Child , Child, Preschool , Endocardial Fibroelastosis/complications , Endocardium/pathology , Female , Heart Ventricles , Humans , Infant , Male
7.
J Biomech Eng ; 143(4)2021 04 01.
Article in English | MEDLINE | ID: mdl-32793949

ABSTRACT

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. Twelve healthy participants practiced modifications in a university hallway directly after performing five normal walking trials. The wearable feedback system provided real-time haptic biofeedback during training trials to inform participants if they were within the prescribed modification range (7-12 deg greater than baseline). Participants were instructed to move to the next modification only once they felt comfortable and could perform it with minimal errors. Following training, five trials of each modification were immediately performed in the gait laboratory without feedback. All participants successfully modified their foot progression and trunk angle using the wearable system. At post-test, PKAM decreased from baseline by 62%, 55%, and 28% during combined, trunk leanand toe-in gait, respectively. The wearable feedback system was effective to modify participants' foot and trunk angle by the prescribed amount, resulting in reduced PKAM during all modifications at post-test. Participants were also able to perform a combined modification, although it took longer to report feeling comfortable doing so. This study demonstrates that a wearable feedback system is feasible to modify kinematic parameters and train gait modifications outside the laboratory.


Subject(s)
Haptic Technology
8.
Pediatr Radiol ; 50(11): 1610-1616, 2020 10.
Article in English | MEDLINE | ID: mdl-32613358

ABSTRACT

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. The morphological features identified on the stereoscopic models were compared with findings at macroscopic examination of the actual heart specimens. The results showed that the mixed-reality system provided highly accurate stereoscopic display of spatially complex congenital cardiac lesions, with interactive features that might enhance 3-D understanding of morphology. Additionally, the authors found that high-resolution digital reproduction of cardiac specimens using clinical CT scanners is feasible for preservation and educational purposes.


Subject(s)
Heart Defects, Congenital/diagnostic imaging , Imaging, Three-Dimensional , Radiographic Image Interpretation, Computer-Assisted/methods , Tomography, X-Ray Computed , Artifacts , Humans , In Vitro Techniques , Proof of Concept Study
9.
Sensors (Basel) ; 20(20)2020 Oct 20.
Article in English | MEDLINE | ID: mdl-33092285

ABSTRACT

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. Furthermore, delta encoding was shown to approach the a posteriori optimal linear compression level. All methods were implemented and released as open source C code using fixed point computation which can be integrated into a variety of computational platforms. These results could serve to inform and enable human movement data compression in a variety of emerging medical and technological applications.


Subject(s)
Data Compression , Movement , Running , Humans , Software , Walking
10.
Am J Med Genet A ; 179(7): 1325-1329, 2019 07.
Article in English | MEDLINE | ID: mdl-31074124

ABSTRACT

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. We present a male infant with ACDMPV, hypoplastic left heart sequence (HLHS), duodenal atresia, and imperforate anus due to a de novo, in frame deletion in FOXF1: c.209_214del (p.Thr70_Leu71del). Previous reports have suggested that microdeletions involving FOXF1 are associated with ACDMPV with congenital heart defects, including HLHS, gastrointestinal atresias, and other anomalies; whereas likely pathogenic variants within FOXF1 have not been reported with ACDMPV and HLHS. This is the first patient reported with ACDMPV, HLHS, imperforate anus, and duodenal atresia associated with a likely pathogenic variant in the FOXF1 gene.


Subject(s)
Forkhead Transcription Factors/genetics , Hypoplastic Left Heart Syndrome/genetics , Persistent Fetal Circulation Syndrome/genetics , Sequence Deletion , Humans , Infant, Newborn , Male
11.
Pediatr Dev Pathol ; 21(6): 528-536, 2018.
Article in English | MEDLINE | ID: mdl-29642737

ABSTRACT

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. Analyzed clinical data included obstetrical history, postpartum/neonatal medical course, and circumstances surrounding death. Evaluated autopsy data included growth parameters, pathological findings, ancillary test results, and COD. Data from decedents in which a COD was established (COD-E) were statistically compared with that from decedents in which the COD was undetermined (COD-U). Of 104 neonates (M: 49; F: 55) who fulfilled our inclusion criteria, a COD was established at autopsy in 46 cases (44%). Infections, congenital abnormalities, and inborn errors of metabolism were the most common CODs. Single variables statistically more likely to be found in COD-E neonates were clinical history of prodromal illness, witnessed loss of vital signs, and evidence of physiological stress in the thymus or the liver. A prodrome was statistically more common in the COD-E group, but the absence of a prodrome does not reliably exclude COD-E cases, since over 50% of these patients were asymptomatic prior to their demise. In COD-U neonates, the statistically significant factors were death during sleep, death during sleep while "bed"-sharing, "heavy" lungs, and petechial hemorrhages on the epicardium or pleura. Given the frequency and wide spectrum of underlying pathologies in COD-E neonates, referral of SUD-N cases to pathologists with specialized pediatric autopsy expertise is recommended.


Subject(s)
Cause of Death , Sudden Infant Death/pathology , Autopsy , Female , Humans , Infant, Newborn , Male , Sudden Infant Death/etiology
12.
Pediatr Crit Care Med ; 19(8S Suppl 2): S69-S71, 2018 08.
Article in English | MEDLINE | ID: mdl-30080813

ABSTRACT

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. DATA EXTRACTION: Moderated by three experts on investigations after death in children. DATA SYNTHESIS: Not relevant. CONCLUSIONS: A multidisciplinary cliniciopathologic conference is important after the death of a child in order to help bring closure to the family and to attempt to address any concerns they may have about the care. It is also an important part of the quality of care process for a tertiary care institution and provides an unique opportunity for ongoing medical education. The model of a multidisciplinary cliniciopathologic conference used by the Ontario Coroner's Office to investigate sudden and unexpected deaths in children under 5 years old, which has been functioning for over 30 years, is described. Reports from this Pediatric Death Review Committee have been influential in improving the care of children in the province of Ontario.


Subject(s)
Autopsy/standards , Cause of Death , Death , Child , Humans , Intensive Care Units, Pediatric/organization & administration , Ontario
13.
New Phytol ; 215(1): 77-84, 2017 Jul.
Article in English | MEDLINE | ID: mdl-28513846

ABSTRACT

Plants sense microbial signatures via activation of pattern recognition receptors (PPRs), which trigger a range of cellular defences. One response is the closure of plasmodesmata, which reduces symplastic connectivity and the capacity for direct molecular exchange between host cells. Plasmodesmal flux is regulated by a variety of environmental cues but the downstream signalling pathways are poorly defined, especially the way in which calcium regulates plasmodesmal closure. Here, we identify that closure of plasmodesmata in response to bacterial flagellin, but not fungal chitin, is mediated by a plasmodesmal-localized Ca2+ -binding protein Calmodulin-like 41 (CML41). CML41 is transcriptionally upregulated by flg22 and facilitates rapid callose deposition at plasmodesmata following flg22 treatment. CML41 acts independently of other defence responses triggered by flg22 perception and reduces bacterial infection. We propose that CML41 enables Ca2+ -signalling specificity during bacterial pathogen attack and is required for a complete defence response against Pseudomonas syringae.


Subject(s)
Arabidopsis Proteins/physiology , Arabidopsis/immunology , Calcium-Binding Proteins/physiology , Calmodulin/physiology , Plasmodesmata/physiology , Arabidopsis/metabolism , Arabidopsis/microbiology , Arabidopsis Proteins/metabolism , Bacterial Proteins/immunology , Calcium Signaling , Calcium-Binding Proteins/metabolism , Calmodulin/metabolism , Cloning, Molecular , Flagellin/immunology , Plasmodesmata/metabolism , Receptors, Pattern Recognition/metabolism , Receptors, Pattern Recognition/physiology
14.
J Cardiovasc Magn Reson ; 19(1): 10, 2017 Feb 01.
Article in English | MEDLINE | ID: mdl-28143545

ABSTRACT

BACKGROUND: Adverse fibrotic remodeling is detrimental to myocardial health and a reliable method for monitoring the development of fibrotic remodeling may be desirable during the follow-up of patients after heart transplantation (HTx). Quantification of diffuse myocardial fibrosis with cardiovascular magnetic resonance (CMR) has been increasingly applied and validated histologically in adult patients with heart disease. However, comparisons of CMR findings with histological fibrosis burden in children are lacking. This study aimed to compare native T1 times and extracellular volume fraction (ECV) derived from CMR with the degree of collagen on endomyocardial biopsy (EmBx), and to investigate the association between myocardial fibrosis and clinical as well as functional markers in children after HTx. METHODS: EmBx and CMR were performed on the same day. All specimens were stained with picrosirius red. The collagen volume fraction (CVF) was calculated as ratio of stained collagen area to total myocardial area on EmBx. Native T1 values and ECV were measured by CMR on a mid-ventricular short axis slice, using a modified look-locker inversion recovery approach. RESULTS: Twenty patients (9.9 ± 6.2 years of age; 9 girls) after HTx were prospectively enrolled, at a median of 1.3 years (0.02-12.6 years) post HTx, and compared to 24 controls (13.9 ± 2.6 years of age; 12 girls). The mean histological CVF was 10.0 ± 3.4%. Septal native T1 times and ECV were higher in HTx patients compared to controls (1008 ± 32 ms vs 979 ± 24 ms, p < 0.005 and 0.30 ± 0.03 vs 0.22 ± 0.03, p < 0.0001, respectively). CVF showed a moderate correlation with native T1 (r = 0.53, p < 0.05) as well as ECV (r = 0.46, p < 0.05). Native T1 time, but not ECV and CVF, correlated with ischemia time (r = 0.46, p < 0.05). CONCLUSIONS: CMR-derived fibrosis markers correlate with histological degree of fibrosis on EmBx in children after HTx. Further, native T1 times are associated with longer ischemia times.


Subject(s)
Cardiomyopathies/diagnostic imaging , Heart Transplantation/adverse effects , Magnetic Resonance Imaging, Cine , Myocardium/pathology , Transplant Recipients , Ventricular Remodeling , Adolescent , Age Factors , Biomarkers/analysis , Biopsy , Cardiomyopathies/etiology , Cardiomyopathies/pathology , Cardiomyopathies/physiopathology , Case-Control Studies , Child , Child, Preschool , Collagen/analysis , Contrast Media/administration & dosage , Female , Fibrosis , Gadolinium DTPA/administration & dosage , Humans , Infant , Male , Myocardium/chemistry , Observer Variation , Pilot Projects , Predictive Value of Tests , Prospective Studies , Reproducibility of Results , Severity of Illness Index , Treatment Outcome
15.
Proc Natl Acad Sci U S A ; 111(13): 4814-9, 2014 Apr 01.
Article in English | MEDLINE | ID: mdl-24707045

ABSTRACT

Glycine max symbiotic ammonium transporter 1 was first documented as a putative ammonium (NH4(+)) channel localized to the symbiosome membrane of soybean root nodules. We show that Glycine max symbiotic ammonium transporter 1 is actually a membrane-localized basic helix-loop-helix (bHLH) DNA-binding transcription factor now renamed Glycine max bHLH membrane 1 (GmbHLHm1). In yeast, GmbHLHm1 enters the nucleus and transcriptionally activates a unique plasma membrane NH4(+) channel Saccharomyces cerevisiae ammonium facilitator 1. Ammonium facilitator 1 homologs are present in soybean and other plant species, where they often share chromosomal microsynteny with bHLHm1 loci. GmbHLHm1 is important to the soybean rhizobium symbiosis because loss of activity results in a reduction of nodule fitness and growth. Transcriptional changes in nodules highlight downstream signaling pathways involving circadian clock regulation, nutrient transport, hormone signaling, and cell wall modification. Collectively, these results show that GmbHLHm1 influences nodule development and activity and is linked to a novel mechanism for NH4(+) transport common to both yeast and plants.


Subject(s)
Ammonium Compounds/metabolism , Basic Helix-Loop-Helix Leucine Zipper Transcription Factors/metabolism , Cation Transport Proteins/metabolism , Glycine max/growth & development , Glycine max/metabolism , Root Nodules, Plant/growth & development , Root Nodules, Plant/metabolism , Soybean Proteins/metabolism , Biological Transport , Cell Membrane/metabolism , DNA, Plant/metabolism , Gene Expression Regulation, Plant , Protein Binding , Root Nodules, Plant/cytology , Root Nodules, Plant/ultrastructure , Saccharomyces cerevisiae/metabolism , Glycine max/genetics , Glycine max/ultrastructure
16.
New Phytol ; 210(3): 1011-21, 2016 May.
Article in English | MEDLINE | ID: mdl-26790563

ABSTRACT

In root nodules rhizobia enter host cells via infection threads. The release of bacteria to a host cell is possible from cell wall-free regions of the infection thread. We hypothesized that the VAMP721d and VAMP721e exocytotic pathway, identified before in Medicago truncatula, has a role in the local modification of cell wall during the release of rhizobia. To clarify the role of VAMP721d and VAMP721e we used Glycine max, a plant with a determinate type of nodule. The localization of the main polysaccharide compounds of primary cell walls was analysed in control vs nodules with partially silenced GmVAMP721d. The silencing of GmVAMP721d blocked the release of rhizobia. Instead of rhizobia-containing membrane compartments - symbiosomes - the infected cells contained big clusters of bacteria embedded in a matrix of methyl-esterified and de-methyl-esterified pectin. These clusters were surrounded by a membrane. We found that GmVAMP721d-positive vesicles were not transporting methyl-esterified pectin. We hypothesized that they may deliver the enzymes involved in pectin turnover. Subsequently, we found that GmVAMP721d is partly co-localized with pectate lyase. Therefore, the biological role of VAMP721d may be explained by its action in delivering pectin-modifying enzymes to the site of release.


Subject(s)
Glycine max/metabolism , Glycine max/microbiology , Pectins/metabolism , Plant Proteins/metabolism , Rhizobium/physiology , Root Nodules, Plant/microbiology , Cellulose/metabolism , Esterification , Gene Silencing , Polysaccharide-Lyases/metabolism , Protein Transport , Root Nodules, Plant/metabolism , Root Nodules, Plant/ultrastructure , Symbiosis
17.
Biochem J ; 457(1): 127-36, 2014 Jan 01.
Article in English | MEDLINE | ID: mdl-24102643

ABSTRACT

Many signalling pathways in plants are regulated by the second messenger calcium (Ca(2+)). In the standard model, Ca(2+)-sensor proteins, such as CaM (calmodulin), detect Ca(2+) signals and subsequently regulate downstream targets to advance the signal transduction cascade. In addition to CaM, plants possess many CMLs (CaM-like proteins) that are predicted to function as Ca(2+) sensors, but which remain largely uncharacterized. In the present study, we examined the biochemical properties, subcellular localization and tissue-specific distribution of Arabidopsis CML43. Our data indicate that CML43 displays characteristics typical of Ca(2+) sensors, including high-affinity Ca(2+) binding, conformational changes upon Ca(2+) binding that expose hydrophobic regions and stabilization of structure in the presence of Mg(2+) or Ca(2+). In vivo localization analysis demonstrates that CML43 resides in cytosolic and nuclear compartments. Transgenic plants expressing a CML43:GUS (ß-glucoronidase) promoter reporter gene revealed that CML43 promoter activity is restricted almost exclusively to root tips under normal growth conditions. GUS reporter activity in these transgenic plants was strongly increased when exposed to the defence compound SA (salicylic acid). Furthermore, immunoblot analysis revealed that the CML43 protein accumulates following treatment with SA. Collectively, our findings suggest that CML43 functions as a Ca(2+) sensor in root tips during both normal growth and plant immune response.


Subject(s)
Arabidopsis Proteins/physiology , Arabidopsis , Calcium-Binding Proteins/physiology , Calcium/metabolism , Gene Expression Regulation, Plant/drug effects , Salicylic Acid/pharmacology , Arabidopsis/drug effects , Arabidopsis/physiology , Calcium-Binding Proteins/chemistry , Calmodulin/genetics , Cells, Cultured , Immune System/metabolism , Organ Specificity/drug effects , Organ Specificity/genetics , Plant Roots/drug effects , Plant Roots/genetics , Plant Roots/metabolism , Plants, Genetically Modified , Sequence Homology , Signal Transduction/drug effects , Signal Transduction/genetics , Nicotiana
18.
Commun Biol ; 6(1): 770, 2023 07 22.
Article in English | MEDLINE | ID: mdl-37481629

ABSTRACT

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. The hypoplastic AAo have miRNA-gene pairs annotating to cell proliferation that are inversely differentially expressed by bulk RNA-seq. Single-nucleus RNA-seq of the hypoplastic LV myocardium shows an increase in fibroblasts with a reciprocal decrease in cardiomyocyte nuclei proportions. Fibroblasts, cardiomyocytes and endothelial cells from hypoplastic myocardium have increased expression of extracellular matrix component or fibrosis genes with dysregulated fibroblast growth factor signaling. Hence, a severe sustained ( ~ 1/3 gestation) reduction in fetal left heart flow is sufficient to cause left heart hypoplasia. This is accompanied by changes in cellular composition and gene expression consistent with a pro-fibrotic environment and aberrant induction of mesenchymal programs.


Subject(s)
Endothelial Cells , Sheep, Domestic , Sheep , Animals , Fetus , Myocardium , Heart Ventricles
19.
Eur Urol Focus ; 8(5): 1560-1563, 2022 09.
Article in English | MEDLINE | ID: mdl-34973956

ABSTRACT

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. During 2000-2013, 19 previously healthy neonates were admitted for acute complications after circumcision. Four were admitted for bleeding, with hemophilia identified in two cases and von Willebrand disease in one. Eight boys required emergency surgery, three for severe bleeding. Four boys with amputation of the glans underwent immediate surgical reconstruction. One infant was taken to the operating room to remove an obstructing Plastibell ring. Seven boys were admitted to the intensive care unit with severe bleeding or sepsis, three of whom ultimately progressed to hemorrhagic or septic shock. Two of these children died of their complications. We estimate that the annual incidence of severe complications requiring hospitalization after NNC in the Greater Toronto Area was approximately 0.01%, and the incidence of fatalities over the 14-yr review period was approximately 0.0012%. Our results indicate that the risk of serious complications and death as a result of NNC is greater than generally assumed.


Subject(s)
Circumcision, Male , Child , Humans , Infant , Infant, Newborn , Male , Circumcision, Male/adverse effects , Databases, Factual , Hemorrhage , Incidence
20.
Pediatr Radiol ; 41(6): 736-48, 2011 Jun.
Article in English | MEDLINE | ID: mdl-21264464

ABSTRACT

BACKGROUND: Studies have reported that thoracic CT may provide greater sensitivity compared with radiography in detection of pediatric rib fractures and fracture healing. The additional sensitivity afforded by thoracic CT may have medicolegal implications where abuse is suspected. OBJECTIVE: To determine the additional value of postmortem thoracic CT compared with radiography in detecting pediatric rib fractures, and fracture healing, using autopsy findings as a gold standard. MATERIALS AND METHODS: We retrospectively reviewed 56 coroner's cases with postmortem radiography and CT thoracic survey. All studies underwent primary interpretation by one or two radiologists. The study radiologist independently reviewed all images from 13 patients with positive findings on radiography, CT or autopsy. Sensitivity and specificity between observers and imaging modalities were compared. RESULTS: Primary interpretation: Fractures were recognized on radiography in 5/12 patients who had fractures found at autopsy, and on CT in 8/12 patients. In total, 29% (24/83) of fractures were reported on radiography, and 51% (52/101) of fractures were reported on CT. Study radiologist: Fractures were recognized on radiography in 7/12 patients who had fractures found at autopsy, and on CT in 11/12 patients. In total, 46% (38/83) of fractures were reported on radiography, and 85% (86/101) of fractures were reported on CT. CONCLUSION: Postmortem thoracic CT provides greater sensitivity than radiography in detecting pediatric rib fractures, most notably in anterior and posterior fractures. However, the degree of improvement in sensitivity provided by CT might depend on observer experience.


Subject(s)
Battered Child Syndrome/diagnostic imaging , Child Abuse/diagnosis , Radiography, Thoracic/methods , Rib Fractures/diagnostic imaging , Tomography, X-Ray Computed/methods , X-Ray Film , Child , Child, Preschool , Female , Humans , Infant, Newborn , Male , Reproducibility of Results , Sensitivity and Specificity
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