Stroke incidence, mortality, subtypes in rural and urban populations in five geographic areas of India (2018-2019): results from the National Stroke Registry Programme.

Background: Increasing stroke burden in India demands a long-term stroke surveillance framework. Earlier studies in India were urban-based, short term and provided limited data on stroke incidence and its outcomes. This gap is addressed by the establishment of five population-based stroke registries (PBSRs) of the National Stroke Registry Programme, India. This paper describes stroke incidence, mortality and age, sex, and subtypes distribution in the five PBSRs with urban and rural populations. Methods: First-ever incident stroke patients in age group ≥18 years, resident for at least one year in the defined geographic area, identified from health facilities were registered. Death records with stroke as the cause of death from the Civil Registration System (CRS) were included. Transient ischemic attack (TIA) was excluded. Three PBSRs (Cuttack, Tirunelveli, Cachar) included urban and rural populations. PBSRs in Kota and Varanasi were urban areas. The crude and age-standardized incidence rate (ASR) by age, sex, and residence (urban and rural), rate ratios of ASR, case fatality proportions and rates at day 28 after onset of stroke were calculated for years 2018-2019. Findings: A total of 13,820 registered first-ever stroke cases that included 985 death certificate-only cases (DCOs) were analysed. The pooled crude incidence rate was 138.1 per 100,000 population with an age-standardized incidence rate (ASR) of 103.4 (both sexes), 125.7 (males) and 80.8 (females). The risk of stroke among rural residents was one in seven (Cuttack), one in nine (Tirunelveli), and one in 15 (Cachar). Ischemic stroke was the most common type in all PBSRs. Age-standardized case fatality rates (ASCFR) per 100,000 population for pooled PBSRs was 30.0 (males) and 18.8 (females), and the rate ratio (M/F) ranged from 1.2 (Cuttack) to 2.0 (Cachar). Interpretation: Population-based registries have provided a comprehensive stroke surveillance platform to measure stroke burden and outcomes by age, sex, residence and subtype across India. The rural-urban pattern of stroke incidence and mortality shall guide health policy and programme planning to strengthen stroke prevention and treatment measures in India. Funding: The National Stroke Registry Programme is funded through the intramural funding of the Indian Council of Medical Research, Department of Health Research, Ministry of Health and Family Welfare, India.

External jasmonic acid isoleucine mediates amplification of plant elicitor peptide receptor (PEPR) and jasmonate-based immune signalling.

Jasmonic acid-isoleucine (JA-Ile) is a plant defence hormone whose cellular levels are elevated upon herbivory and regulate defence signalling. Despite their pivotal role, our understanding of the rapid cellular perception of bioactive JA-Ile is limited. This study identifies cell type-specific JA-Ile-induced Ca2+ signal and its role in self-amplification and plant elicitor peptide receptor (PEPR)-mediated signalling. Using the Ca2+ reporter, R-GECO1 in Arabidopsis, we have characterized a monophasic and sustained JA-Ile-dependent Ca2+ signature in leaf epidermal cells. The rapid Ca2+ signal is independent of positive feedback by the JA-Ile receptor, COI1 and the transporter, JAT1. Microarray analysis identified up-regulation of receptors, PEPR1 and PEPR2 upon JA-Ile treatment. The pepr1 pepr2 double mutant in R-GECO1 background exhibits impaired external JA-Ile induced Ca2+ cyt elevation and impacts the canonical JA-Ile responsive genes. JA responsive transcription factor, MYC2 binds to the G-Box motif of PEPR1 and PEPR2 promoter and activates their expression upon JA-Ile treatment and in myc2 mutant, this is reduced. External JA-Ile amplifies AtPep-PEPR pathway by increasing the AtPep precursor, PROPEP expression. Our work shows a previously unknown non-canonical PEPR-JA-Ile-Ca2+ -MYC2 signalling module through which plants sense JA-Ile rapidly to amplify both AtPep-PEPR and jasmonate signalling in undamaged cells.

Effects of Oral Leak Size on Valsalva Responses.

Introduction: We hypothesized that oral leak size, a hitherto unstudied technical variable, would influence hemodynamic responses and difficulty level in executing the Valsalva maneuver (VM). Methods: Based on power analysis, 38 healthy participants were included. Oral leak size in random order was 0.35, 0.71, 1.01,1.40 mm. Level of difficulty was rated 1 to 10, with ≥7-10 being severe. VM was performed at 40 mmHg expiratory strain for 15 s. Three trials were averaged for each leak size. Data were analyzed by repeated measures ANOVA and pairwise comparisons with Tukey adjustment, a mixed effect model, and a generalized linear model. Results: Of the 38 participants, 4 were excluded from analysis for protocol deviation. Phase II L mean BP (MBP) was significantly higher with the largest leak versus all others (P < 0.001). TL was significantly lower with the largest leak versus all others (P = 0.0029). Difficulty performing the VM increased significantly with every increase in leak size (P < 0.001), and a significantly higher percentage of participants reported severe difficulty with leak 4 compared with leak 1 (P < 0.001), 2 (P = 0.0068), and 3 (P = 0.0068). There was no significant effect of phase II E SBP decline on phase II L MBP increase (P = 0.0752). Difficulty increased significantly with one unit increase in phase II L MBP (P = 0.0002). Conclusions: Oral leak size affected VM hemodynamic parameters and level of difficulty. Oral leak size exceeding 1.01 mm significantly affected phase II L MBP. Level of difficulty rose with increasing leak size and was significantly correlated with increase in phase II L MBP.

Forward Genetic Screen Using Transgenic Calcium Reporter Aequorin to Identify Novel Targets in Calcium Signaling.

Forward genetic screens have been important tools in the unbiased identification of genetic components involved in several biological pathways. The basis of the screen is to generate a mutant population that can be screened with a phenotype of interest. EMS (ethyl methane sulfonate) is a commonly used alkylating agent for inducing random mutation in a classical forward genetic screen to identify multiple genes involved in any given process. Cytosolic calcium (Ca2+) elevation is a key early signaling pathway that is activated upon stress perception. However the identity of receptors, channels, pumps and transporters of Ca2+ is still elusive in many study systems. Aequorin is a cellular calcium reporter protein isolated from Aequorea victoria and stably expressed in Arabidopsis. Exploiting this, we designed a forward genetic screen in which we EMS-mutagenized the aequorin transgenic. The seeds from the mutant plants were collected (M1) and screening for the phenotype of interest was carried out in the segregating (M2) population. Using a 96-well high-throughput Ca2+ measurement protocol, several novel mutants can be identified that have a varying calcium response and are measured in real time. The mutants with the phenotype of interest are rescued and propagated till a homozygous mutant plant population is obtained. This protocol provides a method for forward genetic screens in Ca2+ reporter background and identify novel Ca2+ regulated targets.

Determining the scale at which variation in a single gene changes population yields.

Plant trait diversity is known to influence population yield, but the scale at which this happens remains unknown: divergent individuals might change yields of immediate neighbors (neighbor scale) or of plants across a population (population scale). We use Nicotiana attenuata plants silenced in mitogen-activated protein kinase 4 (irMPK4) - with low water-use efficiency (WUE) - to study the scale at which water-use traits alter intraspecific population yields. In the field and glasshouse, we observed overyielding in populations with low percentages of irMPK4 plants, unrelated to water-use phenotypes. Paired-plant experiments excluded the occurrence of overyielding effects at the neighbor scale. Experimentally altering field arbuscular mycorrhizal fungal associations by silencing the Sym-pathway gene NaCCaMK did not affect reproductive overyielding, implicating an effect independent of belowground AMF interactions. Additionally, micro-grafting experiments revealed dependence on shoot-expressed MPK4 for N. attenuata to vary its yield per neighbor presence. We find that variation in a single gene, MPK4, is responsible for population overyielding through a mechanism, independent of irMPK4's WUE phenotype, at the aboveground, population scale.

Whether on farmland or in a forest, plants do not grow in isolation. Plants compete with their neighbors over limited space and resources, and individual plants respond to this competition in different ways by changing how much they grow and how they use resources. The efficiency with which crop plants use water, for example, is one trait that is dramatically influenced by neighboring plants and is of increasing concern given the warming climate. Understanding the effects of interactions between individual plants in a population as a whole is complicated, especially in natural plant communities where neighbors are often from different species. For this reason, McGale et al. took a different approach and looked at neighbors that were all from the same species and differed only in the activity of a single gene. The species in question was coyote tobacco, a plant that is native to western North America. McGale et al. used genetic engineering to silence a gene called MPK4, which was known from previous studies to have the effect of reducing water-use efficiency. Some of these 'water-inefficient' plants were then grown in mixed populations with plants that had normal levels of MPK4. In experiments conducted both in a glasshouse and at a field station in the Utah desert, McGale et al. found that populations with a low percentage of the MPK4-silenced plants were actually more productive than 'monocultures' that were all one type or the other. Further analysis showed that the increase in productivity did not depend on the different soil nutrient or water use of the different populations, or even the density of the plants in the populations. Pairs of plants grown in single pots essentially ruled out any interactions between immediate neighbors being responsible for the increased productivity, suggesting that that effect must instead emerge at the level of the population. Perhaps unexpectedly, McGale et al. also found that the MPK4-silenced plants and control plants did not actually differ in how they used water when grown in the field (previous studies had all been conducted in glasshouses), indicating that this trait also could not explain the observed population-level effect. Finally, experiments that involved grafting the shoots of one plant onto the roots of another suggested that the effect most likely comes from the aboveground parts of the plant. Ecologists have previously noted that more diverse populations typically have higher productivity. This new finding that a small percentage of slightly different plants in an otherwise uniform population can increase overall productivity will likely to be of special interest to researchers looking to boost the efficiency of agricultural ecosystems. Also, since MPK4 is highly conserved, and thus likely to be found in many plant species, this could be an interesting trait with which to study the interactions of natural plant communities.

Detection of pulmonary embolism on computed tomography: improvement using a model-based iterative reconstruction algorithm compared with filtered back projection and iterative reconstruction algorithms.

PURPOSE: The purpose of the study was to determine whether a model-based iterative reconstruction (MBIR) technique improves diagnostic confidence and detection of pulmonary embolism (PE) compared with hybrid iterative reconstruction (HIR) and filtered back projection (FBP) reconstructions in patients undergoing computed tomography pulmonary angiography. MATERIALS AND METHODS: The study was approved by our institutional review board. Fifty patients underwent computed tomography pulmonary angiography at 100 kV using standard departmental protocols. Twenty-two of 50 patients had studies positive for PE. All 50 studies were reconstructed using FBP, HIR, and MBIR. After image randomization, 5 thoracic radiologists and 2 thoracic radiology fellows graded each study on a scale of 1 (very poor) to 5 (ideal) in 4 subjective categories: diagnostic confidence, noise, pulmonary artery enhancement, and plastic appearance. Readers assessed each study for the presence of PE. Parametric and nonparametric data were analyzed with repeated measures and Friedman analysis of variance, respectively. RESULTS: For the 154 positive studies (7 readers × 22 positive studies), pooled sensitivity for detection of PE was 76% (117/154), 78.6% (121/154), and 82.5% (127/154) using FBP, HIR, and MBIR, respectively. PE detection was significantly higher using MBIR compared with FBP (P = 0.016) and HIR (P = 0.046). Because of nonsignificant increase in FP studies using HIR and MBIR, accuracy with MBIR (88.6%), HIR (87.1%), and FBP (87.7%) was similar. Compared with FBP, MBIR led to a significant subjective increase in diagnostic confidence, noise, and enhancement in 6/7, 6/7, and 7/7 readers, respectively. Compared with HIR, MBIR led to significant subjective increase in diagnostic confidence, noise, and enhancement in 5/7, 5/7, and 7/7 readers, respectively. MBIR led to a subjective increase in plastic appearance in all 7 readers compared with both FBP and HIR. CONCLUSIONS: MBIR led to significant increase in PE detection compared with FBP and HIR. MBIR led to qualitative improvements in diagnostic confidence, perceived noise, and perceived enhancement compared with FBP and HIR.

Valsalva maneuver: shortest optimal expiratory strain duration.

PURPOSE: To quantitate the level of difficulty and determine consistency of hemodynamic responses with various expiratory strain (ES) durations. METHODS: Thirty-four healthy subjects performed the Valsalva maneuver (VM) with an ES duration of 10, 12, and 15 seconds in random order. Level of difficulty after each trial was rated 1 to 10, with 10 being the most difficult. Blood pressure and heart rate (HR) were recorded continuously and non-invasively. Parameters studied were Valsalva ratio (VR), early phase II (IIE), late phase II (IIL), tachycardia latency (TL), bradycardia latency (BL), and overshoot latency (OV-L). Consistency of responses was calculated. RESULTS: DIFFICULTY INCREASED SIGNIFICANTLY WITH INCREASED ES DURATION: 5.1±0.1 (mean±SEM) at 10 seconds, 5.9±0.1 at 12 seconds, and 6.8±0.1 at 15 seconds (p<0.001). Phase IIE, TL, BL, OV-L, and VR response did not differ statistically with increasing ES durations, and there were no differences in variability. Phase IIL response increased significantly with increasing ES duration. Phase IIL was poorly delineated in 14 of 102 trials with 10 seconds ES duration. CONCLUSIONS: ES duration of 10 seconds created a low level of difficulty in healthy individuals. This strain duration produced consistent hemodynamic response for all parameters tested except IIL phase. The absence of IIL phase with 10 seconds ES should not be interpreted as an indicator of sympathetic vasoconstrictor failure.