OsJAZ11 regulates spikelet and seed development in rice.

Seed size is one of the major determinants of seed weight and eventually, crop yield. As the global population is increasing beyond the capacity of current food production, enhancing seed size is a key target for crop breeders. Despite the identification of several genes and QTLs, current understanding about the molecular regulation of seed size/weight remains fragmentary. In the present study, we report novel role of a jasmonic acid (JA) signaling repressor, OsJAZ11 controlling rice seed width and weight. Transgenic rice lines overexpressing OsJAZ11 exhibited up to a 14% increase in seed width and ~30% increase in seed weight compared to wild type (WT). Constitutive expression of OsJAZ11 dramatically influenced spikelet morphogenesis leading to extra glume-like structures, open hull, and abnormal numbers of floral organs. Furthermore, overexpression lines accumulated higher JA levels in spikelets and developing seeds. Expression studies uncovered altered expression of JA biosynthesis/signaling and MADS box genes in overexpression lines compared to WT. Yeast two-hybrid and pull-down assays revealed that OsJAZ11 interacts with OsMADS29 and OsMADS68. Remarkably, expression of OsGW7, a key negative regulator of grain size, was significantly reduced in overexpression lines. We propose that OsJAZ11 participates in the regulation of seed size and spikelet development by coordinating the expression of JA-related, OsGW7 and MADS genes.

OsJAZ11 regulates phosphate starvation responses in rice.

MAIN CONCLUSION: OsJAZ11 regulates phosphate homeostasis by suppressing jasmonic acid signaling and biosynthesis in rice roots. Jasmonic Acid (JA) is a key plant signaling molecule which negatively regulates growth processes including root elongation. JAZ (JASMONATE ZIM-DOMAIN) proteins function as transcriptional repressors of JA signaling. Therefore, targeting JA signaling by deploying JAZ repressors may enhance root length in crops. In this study, we overexpressed JAZ repressor OsJAZ11 in rice to alleviate the root growth inhibitory action of JA. OsJAZ11 is a low phosphate (Pi) responsive gene which is transcriptionally regulated by OsPHR2. We report that OsJAZ11 overexpression promoted primary and seminal root elongation which enhanced Pi foraging. Expression studies revealed that overexpression of OsJAZ11 also reduced Pi starvation response (PSR) under Pi limiting conditions. Moreover, OsJAZ11 overexpression also suppressed JA signaling and biosynthesis as compared to wild type (WT). We further demonstrated that the C-terminal region of OsJAZ11 was crucial for stimulating root elongation in overexpression lines. Rice transgenics overexpressing truncated OsJAZ11ΔC transgene (i.e., missing C-terminal region) exhibited reduced root length and Pi uptake. Interestingly, OsJAZ11 also regulates Pi homeostasis via physical interaction with a key Pi sensing protein, OsSPX1. Our study highlights the functional connections between JA and Pi signaling and reveals JAZ repressors as a promising candidate for improving low Pi tolerance of elite rice genotypes.

OsJAZ9 is involved in water-deficit stress tolerance by regulating leaf width and stomatal density in rice.

Drought stress poses a severe threat to grain yield in rice. Our previous report demonstrated the role of OsJAZ9 in potassium homeostasis by modulating Jasmonic Acid (JA) signalling. While both potassium (K) and JA are known to have an important role in drought stress response, JA's repressor, i.e., JAZs' role in drought stress, remains elusive. Here we report that OsJAZ9 plays a critical role in rice water-deficit stress tolerance via influencing JA and ABA signalling. Overexpression of OsJAZ9 led to the enhanced ABA and JA levels. Our data further revealed that exogenous JA application antagonises the ABA-mediated inhibition of seed germination. Further, OsJAZ9 overexpression reduces leaf width and stomata density, leading to lower leaf transpiration rates than WT. This reduced transpiration and higher K content as osmoticum improved the water-deficit stress tolerance in OsJAZ9 overexpression lines. On the contrary, OsJAZ9 RNAi lines displayed enhanced sensitivity towards water-deficit stress. Our data provide new insights on the role of JA signalling repressors in rice response to water-deficit stress.

OsJAZ9 overexpression modulates jasmonic acid biosynthesis and potassium deficiency responses in rice.

KEY MESSAGE: Enhanced bioactive JA (JA-Ile) accumulation in OsJAZ9 overexpressing rice helps plants tolerate K deficiency. Potassium (K) represents up to 10% of the plant's total dry biomass, and its deficiency makes plants highly susceptible to both abiotic and biotic stresses. K shortage results in the inhibition of root and shoots growth, but the underlying mechanism of this response is unclear. Our RNA-Seq and qPCR analysis suggested leading roles for JA pathway genes under K deficiency in rice. Notably, K deficiency and JA application produced similar phenotypic and transcriptional responses. Here, we integrated molecular, physiological and morphological studies to analyze the role of OsJAZ9 in JA homeostasis and K deficiency responses. We raised OsJAZ9 over-expression, knockdown, transcriptional reporter, translational reporter and C-terminal deleted translational reporter lines in rice to establish the role of JA signaling in K ion homeostasis. JA profiling revealed significantly increased JA-Ile levels in OsJAZ9 OE lines under K deficiency. Furthermore, we established that OsJAZ9 overexpression and knockdown result in K deficiency tolerance and sensitivity, respectively, by modulating various K transporters and root system architecture. Our data provide evidence on the crucial roles of OsJAZ9 for improving K deficiency tolerance in rice by altering JA levels and JA responses.

Heat Treatment, Impact Properties, and Fracture Behaviour of Ti-6Al-4V Alloy Produced by Powder Compact Extrusion.

The mechanical properties of titanium and titanium alloys are very sensitive to processing, microstructure, and impurity levels. In this paper, a blended powder mixture of Ti-6Al-4V alloy was consolidated by powder compact extrusion that involved warm compaction, vacuum sintering, and hot extrusion. The as-processed material with an oxygen content of 0.34 wt.% was subjected to various annealing treatments. The impact toughness of heat-treated material was determined using Charpy V-notch impact testing at room temperature. An emphasis was placed on establishing a relationship among fracture behaviour, microstructure, and the resulting properties of tested material. From the results, it is apparent that the highest impact toughness value of 19.3 J was achieved after α/ß annealing and is comparable with typical values given in the literature for wrought Ti-6Al-4V. In terms of fracture behaviour, it is quite apparent that the crack propagation behaviour of powder-produced material is rather complex compared with the limited amount of data reported for ingot counterparts.

Safety and immunogenicity of a killed bivalent (O1 and O139) whole-cell oral cholera vaccine in adults and children in Vellore, South India.

This open-label study assessed the safety and immunogenicity of two doses (14 days apart) of an indigenously manufactured, killed, bivalent (Vibrio cholerae O1 and O139), whole-cell oral cholera vaccine (SHANCHOL; Shantha Biotechnics) in healthy adults (n = 100) and children (n = 100) in a cholera endemic area (Vellore, South India) to fulfill post-licensure regulatory requirements and post-World Health Organization (WHO) prequalification commitments. Safety and reactogenicity were assessed, and seroconversion rates (i.e. proportion of participants with a ≥ 4-fold rise from baseline in serum vibriocidal antibody titers against V. cholerae O1 Inaba, O1 Ogawa and O139, respectively) were determined 14 days after each vaccine dose. No serious adverse events were reported during the study. Commonly reported solicited adverse events were headache and general ill feeling. Seroconversion rates after the first and second dose in adults were 67.7% and 55.2%, respectively, against O1 Inaba; 47.9% and 45.8% against O1 Ogawa; and 19.8% and 20.8% against O139. In children, seroconversion rates after the first and second dose were 80.2% and 68.8%, respectively, against O1 Inaba; 72.9% and 67.7% against O1 Ogawa; and 26.0% and 18.8% against O139. The geometric mean titers against O1 Inaba, O1 Ogawa, and O139 in both adults and children were significantly higher after each vaccine dose compared to baseline titers (P < 0.001; for both age groups after each dose versus baseline). The seroconversion rates for O1 Inaba, O1 Ogawa, and O139 in both age groups were similar to those in previous studies with the vaccine. In conclusion, the killed, bivalent, whole-cell oral cholera vaccine has a good safety and reactogenicity profile, and is immunogenic in healthy adults and children. Trial Registration: ClinicalTrials.gov NCT00760825; CTRI/2012/01/002354.

Differential expression of salt-responsive genes to salinity stress in salt-tolerant and salt-sensitive rice (Oryza sativa L.) at seedling stage.

The understanding of physio-biochemical and molecular attributes along with morphological traits contributing to the salinity tolerance is important for developing salt-tolerant rice (Oryza sativa L.) varieties. To explore these facts, rice genotypes CSR10 and MI48 with contrasting salt tolerance were characterized under salt stress (control, 75 and 150 mM NaCl) conditions. CSR10 expressed higher rate of physio-biochemical parameters, maintained lower Na/K ratio in shoots, and restricted Na translocation from roots to shoots than MI48. The higher expression of genes related to the osmotic module (DREB2A and LEA3) and ionic module (HKT2;1 and SOS1) in roots of CSR10 suppresses the stress, enhances electrolyte leakage, promotes the higher compatible solute accumulation, and maintains cellular ionic homeostasis leading to better salt stress tolerance than MI48. This study further adds on the importance of these genes in salt tolerance by comparing their behaviour in contrasting rice genotypes and utilizing specific marker to identify salinity-tolerant accessions/donors among germplasm; overexpression of these genes which accelerate the selection procedure precisely has been shown.

Identification of Purple Acid Phosphatases in Chickpea and Potential Roles of CaPAP7 in Seed Phytate Accumulation.

Purple acid phosphatases (PAPs) play important roles in phosphate (Pi) acquisition and utilization. These PAPs hydrolyze organic Phosphorus (P) containing compounds in rhizosphere as well as inside the plant cell. However, roles of PAPs in one of the most widely cultivated legumes, chickpea (Cicer arietnum L.), have not been unraveled so far. In the present study, we identified 25 putative PAPs in chickpea (CaPAPs) which possess functional PAP motifs and domains. Differential regulation of CaPAPs under different nutrient deficiencies revealed their roles under multiple nutrient stresses including Pi deficiency. Interestingly, most of the CaPAPs were prominently expressed in flowers and young pods indicating their roles in flower and seed development. Association mapping of SNPs underlying CaPAPs with seed traits revealed significant association of low Pi inducible CaPAP7 with seed weight and phytate content. Biochemical characterization of recombinant CaPAP7 established it to be a functional acid phosphatase with highest activity on most abundant organic-P substrate, phytate. Exogenous application of recombinant CaPAP7 enhanced biomass and Pi content of Arabidopsis seedlings supplemented with phytate as sole P source. Taken together, our results uncover the PAPs in chickpea and potential roles of CaPAP7 in seed phytate accumulation.

DNA Ploidy and S-phase Fraction Analysis in Paediatric B-cell Acute Lymphoblastic Leukemia Cases: a Tertiary Care Centre Experience.

DNA ploidy is an important prognostic parameter in paediatric B-ALL, but the significance of the S-phase fraction is unclear. In present study, DNA ploidy was assessed in 40 pediatric B-ALL cases by flow cytometry. The DI (DNA index) and percentage of cells in S-phase were calculated using Modfit software. Aneuploidy was noted in 26/40 (65%) cases. A DI of 1.10-1.6 (hyperdiploidy B) was noted in 20/40 (50%) and 6/40 (15%) had a DI>1.60 (triploid and tetraploid range). Some 14/40 (35%) cases had a diploid DI between 0.90-1.05. None of the cases had a DI <0.90 (hypodiploid) or in the 1.06-1.09 (hyperdiploid A) range. The mean S-phase fraction was 2.6%, with 24/40 (60%) having low and 16/40 (40%) high S-phase fractions. No correlation was noted with standard ALL risk and treatment response factors with DI values or S-phase data, except for a positive correlation of low S-phase with high NCI risk category (p=0.032). Overall frequency of hyperdiploidy in our cohort of B-ALL patients was very high (65%). No correlation between hyperdiploidy B and low TLC or common B-phenotype was observed in our study as 42% cases with DI 1.10-1.6 had TLC> 50 x 109 and 57.1% CD 10 negativity. The study also highlighted that S-phase fraction analysis does not add any prognostic information and is not a useful parameter for assessment in ALL cases. However, larger studies with long term outcome analysis are needed to derive definitive conclusions.

A randomized, non-inferiority trial comparing two bivalent killed, whole cell, oral cholera vaccines (Euvichol vs Shanchol) in the Philippines.

BACKGROUND: Currently, there are two oral cholera vaccines (OCV) that are prequalified by the World Health Organization. Both (Dukoral and Shanchol) have been proven to be safe, immunogenic, and effective. As the global supply of OCV remains limited, we assessed the safety and immunogenicity of a new low cost, killed, bivalent OCV (Euvichol) in the Philippines. METHODS: The randomized controlled trial was carried out in healthy Filipino adults and children. Two doses of either the current WHO prequalified OCV (Shanchol) or the same composition OCV being considered for WHO prequalification (Euvichol) were administered to participants. RESULTS: The pivotal study was conducted in total of 1263 healthy participants (777 adults and 486 children). No serious adverse reactions were elicited in either vaccine groups. Vibriocidal antibody responses to V. cholerae O1 Inaba following administration of two doses of Euvichol were non-inferior to those of Shanchol in adults (82% vs 76%) and children (87% vs 89%). Similar findings were observed for O1 Ogawa in adults (80% vs 74%) and children (91% vs 88%). CONCLUSION: A two dose schedule with Euvichol induces a strong vibriocidal response comparable to those elicited by the currently WHO prequalified OCV, Shanchol. Euvichol will be an oral cholera vaccine suitable for use in lower income countries, where cholera still has a significant economic and public health impact.

A Randomized, Placebo-Controlled Trial Evaluating Safety and Immunogenicity of the Killed, Bivalent, Whole-Cell Oral Cholera Vaccine in Ethiopia.

Killed whole-cell oral cholera vaccine (OCV) has been a key component of a comprehensive package including water and sanitation measures for recent cholera epidemics. The vaccine, given in a two-dose regimen, has been evaluated in a large number of human volunteers in India, Vietnam, and Bangladesh, where it has demonstrated safety, immunogenicity, and clinical efficacy. We conducted a double-blind randomized placebo-controlled trial in Ethiopia, where we evaluated the safety and immunogenicity of the vaccine in 216 healthy adults and children. OCV was found to be safe and elicited a robust immunological response against Vibrio cholerae O1, with 81% adults and 77% children demonstrating seroconversion 14 days after the second dose of vaccine. This is the first study to evaluate safety and immunogenicity of the vaccine in a population outside Asia using a placebo-controlled, double-blind, randomized study design.

Angiographic profile and endovascular interventions in Takayasu's arteritis.

BACKGROUND: Takayasu's arteritis (TA) is a rare inflammatory, granulomatous vasculitis primarily involving the aorta and its major branches. Clinical and angiographic manifestations of the disease differ in various parts of the world. OBJECTIVES: To analyze the clinical features, angiographic profile, and endovascular interventions in TA patients (pts). METHODS: From January 2009 to March 2013, a total of 62 consecutive pts who were diagnosed with Takayasu's arteritis at our institute based on American College of Rheumatology Criteria were included in the study. Thirty-four pts underwent conventional angiography, while 28 were evaluated using computed tomographic angiography. Endovascular interventions were attempted for 24 lesions in 23 pts who were symptomatic and had significant stenotic lesions. RESULTS: The mean age of presentation was 28.4 ± 9.7 years, with 69.4% female. Limb claudication (52%) was the most common presenting symptom, followed by vertigo (35%) and constitutional symptoms. The most common angiography class was type V (37.1%), followed by type I (32.2%), type IV (17.7%), type IIa (8.1%), type III (3.2%), and type IIb (1.6%). The most common artery involved was the subclavian (64.5%), with the left subclavian more commonly involved than the right, followed by the abdominal aorta (51.6%) and renal artery (32.2%). Stenotic lesions were present in 59 pts (95%), while aneurysmal involvement was seen in 17 (27.4%), with isolated aneurysmal involvement in 3 of these. CONCLUSIONS: Our cohort of TA pts had more severe and widespread involvement. Despite a more advanced presentation, percutaneous balloon angioplasty with stent implantation is feasible, with good immediate results but slightly higher complication rates.

Idiopathic dilatation of the pulmonary artery-a case report.

Idiopathic dilatation of the pulmonary artery (IDPA) is a rare anomaly presenting with enlargement of the pulmonary artery in the absence of an identifiable cause. The natural history of this silent but potentially life-threatening disease can be unpredictable. We report a case of IDPA in an 80-year-old female with a massively dilated pulmonary artery.

Marfan syndrome with spontaneous rupture of aneurysm of common iliac artery.

Marfan syndrome rarely presents with peripheral artery and pulmonary artery aneurysms. We are presenting a case of a fifteen-year-old boy who presented to us with progressive lower abdominal pulsatile swelling with bruit in the right inguinal region for one month. He had typical marfanoid habitus, though there was no history of similar illness in family. CT angiogram revealed the presence having giant right common iliac aneurysm along with interlobar pulmonary artery aneurysm. He had spontaneous rupture of aneurysm in right common iliac artery.

Transjugular approach for successful recanalization and stenting for inferior vena cava stenosis.

Budd-Chiari syndrome is a rare disease characterized by obstruction of outflow in the hepatic vein and/or the inferior vena cava (IVC). Percutaneous transluminal angioplasty and stent placement is nowadays considered to be the first-line treatment for central venous disease because of its minimal-invasive approach. IVC reconstruction by surgical approach is not preferred due to increased morbidity and disappointing patency rates. We describe a case of a long-segment, thrombotic, chronic total occlusion of the IVC that was dilated and stented using a recanalization technique involving the use of Brokenborough septal puncture needle, Mullin dilator and Accura balloon from the jugular approach.