Siderophore of plant growth promoting rhizobacterium origin reduces reactive oxygen species mediated injury in Solanum spp. caused by fungal pathogens.

AIMS: The study aims to explore antifungal properties of bacillibactin siderophore produced by the plant growth-promoting rhizobacterium (PGPR) Bacillus subtilis against fungal phytopathogens Alternaria porri and Fusarium equiseti isolated from Solanum lycopersicum and Solanum melongena plants. METHODS AND RESULTS: Alternaria porri and F. equiseti were isolated from infected plants of eggplant and tomato, respectively. A plate assay was employed to assess the effect of bacillibactin against the phytopathogens. The antifungal potential of the PGPR was evaluated by estimation of dry fungal biomass, visualization of cellular deformity using compound and scanning electron microscopy, antioxidative enzyme assay and analysis of membrane damage via using lipid peroxidation. Inductively coupled plasma atomic emission spectroscopy (ICP-AES) analysis was employed to investigate changes in intracellular iron content. The impact of bacillibactin on pathogenesis was evaluated by infecting detached leaves of S. lycopersicum and S. melongena plants with both the pathogens and treating the infected leaves with bacillibactin. Leaves were further investigated for ROS accumulation, extent of necrosis and cell death. Our findings revealed significant damage to the hyphal structure of A. porri and F. equiseti following treatment with bacillibactin. Biomass reduction, elevated antioxidative enzyme levels, and membrane damage further substantiated the inhibitory effects of the siderophore on fungal growth. ICP-AES analysis indicates an increase in intracellular iron content suggesting enhanced iron uptake facilitated by bacillibactin. Moreover, application of 1500 µg ml-1 bacillibactin on infected leaves demonstrated a substantial inhibition of ROS accumulation, necrosis, and cell death upon bacillibactin treatment. CONCLUSIONS: This study confirms the potent antagonistic activity of bacillibactin against both the phytopathogens A. porri and F. equiseti growth, supporting its potential as a promising biological control agent for fungal plant diseases. Bacillibactin-induced morphological, physiological, and biochemical alterations in the isolated fungi and pathogen-infected leaves highlight the prospects of bacillibactin as an effective and sustainable solution to mitigate economic losses associated with fungal infections in vegetable crops.

Dissection of QTLs conferring drought tolerance in B. carinata derived B. juncea introgression lines.

BACKGROUND: Drought is one of the important abiotic stresses that can significantly reduce crop yields. In India, about 24% of Brassica juncea (Indian mustard) cultivation is taken up under rainfed conditions, leading to low yields due to moisture deficit stress. Hence, there is an urgent need to improve the productivity of mustard under drought conditions. In the present study, a set of 87 B. carinata-derived B. juncea introgression lines (ILs) was developed with the goal of creating drought-tolerant genotypes. METHOD: The experiment followed the augmented randomized complete block design with four blocks and three checks. ILs were evaluated for seed yield and its contributing traits under both rainfed and irrigated conditions in three different environments created by manipulating locations and years. To identify novel genes and alleles imparting drought tolerance, Quantitative Trait Loci (QTL) analysis was carried out. Genotyping-by-Sequencing (GBS) approach was used to construct the linkage map. RESULTS: The linkage map consisted of 5,165 SNP markers distributed across 18 chromosomes and spanning a distance of 1,671.87 cM. On average, there was a 3.09 cM gap between adjoining markers. A total of 29 additive QTLs were identified for drought tolerance; among these, 17 (58.6% of total QTLs detected) were contributed by B. carinata (BC 4), suggesting a greater contribution of B. carinata towards improving drought tolerance in the ILs. Out of 17 QTLs, 11 (64.7%) were located on the B genome, indicating more introgression segments on the B genome of B. juncea. Eight QTL hotspots, containing two or more QTLs, governing seed yield contributing traits, water use efficiency, and drought tolerance under moisture deficit stress conditions were identified. Seventeen candidate genes related to biotic and abiotic stresses, viz., SOS2, SOS2 like, NPR1, FAE1-KCS, HOT5, DNAJA1, NIA1, BRI1, RF21, ycf2, WRKY33, PAL, SAMS2, orf147, MAPK3, WRR1 and SUS, were reported in the genomic regions of identified QTLs. CONCLUSIONS: The significance of B. carinata in improving drought tolerance and WUE by introducing genomic segments in Indian mustard is well demonstrated. The findings also provide valuable insights into the genetic basis of drought tolerance in mustard and pave the way for the development of drought-tolerant varieties.

Immunometabolic impact of pancreastatin inhibitor PSTi8 in MCD induced mouse model of oxidative stress and steatohepatitis.

AIM: Pancreastatin, a dysglycemic hormone that encourages inflammation and steatosis in a variety of metabolic disorder animal models. The purpose of this study is to determine the effect of the pancreastatin inhibitor PSTi8 on immunometabolic changes in the liver of MCD-induced NASH mice. MAIN METHODS: Methionine and choline-deficient (MCD) diet was used for the development of NASH. Liver enzymes like SGOT, SGPT, and ALP and lipid profiles were also performed in the serum. Further, immunophenotyping study was performed in the liver through flowcytometer. Subsequently, Hematoxylin and Eosin, Picro Sirius Red and Masson's Trichrome staining were done to check the liver morphology and collagen staining, respectively. Inflammatory cytokines were measured through ELISA and gene expression through RT-PCR. The expression of α-SMA was examined using immunohistochemistry and immunofluorescence staining. KEY FINDINGS: PSTi8 inhibited the expression of lipogenic genes in the liver and attenuated bad cholesterol, SGOT, SGPT, and ALP in the serum. PSTi8 improved the liver morphology and attenuated collagen deposition. Subsequently, PSTi8 attenuated inflammatory M1-macrophages, CD8+T, CD4+T cells and increased anti-inflammatory M2 macrophages, T-reg and eosinophil populations in the liver. It also attenuated the expression of pro-inflammatory genes like Mcp1, Tnfα, and Il6. Apart from this, PSTi8 attenuated the oxidative stress marker, like ROS, and MDA and fibrosis marker α-SMA in the liver. It also decreased the apoptosis and ROS and MDA level in the liver. SIGNIFICANCE: Overall, these compressive studies revealed that PSTi8 exhibited beneficial effect on the liver of MCD-induced NASH mice by attenuating inflammation and oxidative stress.

Pancreastatin inhibitor PSTi8 ameliorates insulin resistance by decreasing fat accumulation and oxidative stress in high-fat diet-fed mice.

Abnormal fat accumulation, enhanced free fatty acids (FFA) release, and their metabolites cause insulin resistance (IR) in major glucose-lipid metabolic organs such as skeletal muscle and adipose tissue. However, excessive lipolysis and FFA release from adipose tissue elevate plasma FFA levels leading to oxidative stress and skeletal muscle IR. Indeed, in obese individuals, there is enhanced pro-inflammatory secretion from adipose tissue influencing insulin signaling in skeletal muscles. Here, we investigated the effect of PSTi8 on FFA-induced IR in both in vitro and in vivo models. Palmitate (Pal)-treated 3T3-L1 cells increased lipid accumulation as well as lipolysis, which reduced the insulin-stimulated glucose uptake. PSTi8 treatment significantly prevented Pal-induced lipid accumulation, and release and enhanced insulin-stimulated glucose uptake. It further reduced the release of pro-inflammatory cytokines from Pal-treated 3T3-L1 cells as well as from adipose tissue explants. In addition, PSTi8 treatment decreases M1 surface markers in Pal-treated bone marrow-derived monocytes (BMDM). PSTi8 treatment also significantly enhanced the Pal-mediated reduced skeletal muscle glucose disposal and reduced intracellular oxidative stress. In vitro effect of PSTi8 was consistent with in vivo HFD-fed mice IR model. PSTi8 treatment in HFD-fed mice significantly improved glucose metabolism and enhanced skeletal muscle insulin sensitivity with reduced adiposity and pro-inflammatory cytokines. Taken together, our results support that PSTi8 treatment can protect both adipose and skeletal muscles from FFA-induced IR.

Temporal changes in sleep quality and knee function following primary total knee arthroplasty: a prospective study.

PURPOSE: Several patient-reported outcome measures (PROMs) have been used to assess improvement in the quality of life following total knee arthroplasty (TKA). However, there is paucity of studies evaluating the sleep quality and knee function following TKA. The primary aim of our study was to evaluate the sleep quality and knee function in primary TKA patients using the Pittsburgh Sleep Quality Index (PSQI) and Knee Society Score (KSS), respectively. The secondary aim was to assess the correlation between the two outcome measures over the course of first post-operative year following TKA. METHODS: One hundred sixty-eight patients (female-140/male-28) with mean age of 64.63 years (± 7.50) who underwent 168 primary unilateral TKA using a cemented posterior-stabilised implant without patella resurfacing between June 2018 and October 2018 were included in the study. Global PSQI and KSS were recorded pre-operatively and post-operatively weekly up to six weeks and at one year. Body mass index (BMI) and Charlson comorbidity index (CCI) were recorded during pre-operative assessment. RESULTS: Mean(± SD) BMI and CCI were 28.45(± 4.64) and 2.48(± 0.93), respectively. Pre-operative global PSQI of 1.98(± 0.97) increased to 13.48(± 3.36) in the first post-operative week (p < 0.001) and remained high during all the six weeks following TKA (p < 0.001), whereas at the first post-operative year, it reduced to 2.10(± 1.15) (p = 0.15). Pre-operative KSS of 52.00(± 9.98) increased to 71.67(± 6.58) and 85.49(± 4.67) at 6 weeks and the first post-operative year respectively (p < 0.001). Pre-operative global PSQI had moderate correlation with pre-operative KSS (r = 0.39) (p < 0.001). Strong correlation was noted between pre-operative global PSQI and six week post-operative KSS (r = 0.47) (p < 0.001). Low correlation was noted between pre-operative global PSQI and KSS at the first post-operative year (r = 0.10, p = 0.19) following TKA. Multiple regression analysis revealed age, CCI, and pre-operative range of motion as independent predictors of global PSQI. CONCLUSIONS: Patients undergoing TKA experience changes in sleep quality but report an overall improvement in knee function during the first post-operative year. Sleep quality has moderate to strong correlation with knee function in the early post-operative period beyond which there is a low correlation with knee function thereby suggesting a transient phenomenon. Hence patients undergoing TKA can be appropriately counselled regarding the variation in sleep quality in the post-operative period and reassured accordingly.

Biofilm Inhibition and Antiviral Response of Cold Sprayed and Shot Peened Copper Surfaces: Effect of Surface Morphology and Microstructure.

Antibacterial properties of copper against planktonic bacteria population are affected by surface microstructure and topography. However, copper interactions with bacteria in a biofilm state are less studied. This work aims at better understanding the difference in biofilm inhibition of bulk, cold-sprayed, and shot-peened copper surfaces and gaining further insights on the underlying mechanisms using optical and scanning electron microscopy to investigate the topography and microstructure of the surfaces. The biofilm inhibition ability is reported for all surfaces. Results show that the biofilm inhibition performance of cold sprayed copper, while initially better, decreases with time and results in an almost identical performance than as-received copper after 18h incubation time. The shot-peened samples with a rough and ultrafine microstructure demonstrated an enhanced biofilm control, especially at 18 hr. The biofilm control mechanisms were explained by the diffusion rates and concentration of copper ions and the interaction between these ions and the biofilm, while surface topography plays a role in the bacteria attachment at the early planktonic state. Furthermore, the data suggest that surface topography plays a key role in antiviral activity of the materials tested, with a smooth surface being the most efficient.

Anomalous left hepatic vein to coronary sinus in a patient with atrial septal defect: Minimally invasive approach; technical challenges.

Left hepatic vein draining into coronary sinus is a rare systemic vascular anomaly. Its presence is significant when it is associated with other cardiac lesions requiring surgery. We report technical challenges in a case of persistent left superior vena cava and left hepatic vein draining into coronary sinus in an adult with ostium secundum atrial septal defect, which was repaired through minimally invasive approach. The main technical challenge in this case was to achieve adequate venous drainage, which was achieved by vacuum assistance and by manipulating the position of femoral venous cannula. We approached through a right anterolateral thoracotomy and adequate venous drainage was achieved without cannulating left hepatic vein or left superior vena cava.

Utility and relevance of modified lateral trapezius myocutaneous flap as a locoregional reconstructive option for medium-sized ablative defects in head and neck cancer-our experience.

PURPOSE: Lateral trapezius myocutaneous (LTMC) flap has been used less frequently for defects pertaining to head and neck cancer ablative surgical defects. This study is to assess the utility of LTMC flap with a new modification of the flap design. METHODS: This is a retrospective study using LTMC flap with modifications as a reconstructive option for intraoral & extraoral skin defects in head and neck cancer patients following ablative surgery, from August 2014 to October 2019 in a tertiary care hospital. The original technique of LTMC flap was modified for better results and outcomes. RESULTS: Thirty-five patients underwent lateral trapezius flap reconstruction for ablative defects of oral cavity, laryngopharynx, neck and parotid. Twenty-three patients (65.71%) underwent adjuvant radiation/radiation-chemotherapy while one patient defaulted. Two patients (5.71%) had major flap failure. After the loss of flap in these patients, the operative technique was further modified for better results. However, the disadvantage of using this flap is shoulder disability. Nonetheless in the present study, the disability was reduced after modifying the operating technique by preserving the spinal accessory nerve. CONCLUSION: The present study demonstrated minimal functional loss, low morbidity with satisfactory results, thus also reducing treatment cost. Thus, LTMC is a reliable reconstructive option in head and neck cancer patients for medium-sized defects.

Chemometric approach based characterization and selection of mid-early cauliflower for bioactive compounds and antioxidant activity.

The present study was aimed to analyse bioactive compounds (total phenolics, ascorbic acid and sinigrin) and antioxidant activity in 14 mid-early cauliflower genotypes. Significant differences (pb 0.05) were observed among the genotypes for all bioactive compounds and antioxidant activity. Total phenolics content of curd were ranged from 20.36 to 48.93 mg gallic acid equivalent (GAE) 100 g-1 fresh weight (FW) which showed 2.5 times variation. The ascorbic acid content was maximum in DC522 (88.53 mg 100 g-1 FW) followed by Pusa Sharad (65.64 mg 100 g-1 FW) while minimum in DC310 (39.62 65.64 mg 100 g-1 FW). Wide variation was observed for cupric reducing antioxidant capacity and ferric reducing antioxidant power ranging from 9.04 to 20.83 mg GAE 100 g-1 FW and 13.11 to 26.31 mg GAE 100 g-1 FW, respectively. Sinigrin was found to be highest in DC306 (39.50 µmol 100 g-1 FW) for leaf and in DC326 (36.93 µmol 100 g-1 FW) for curd sample. The cauliflower genotypes were classified based on chemometric approaches namely principal component analysis (PCA) and agglomerative hierarchical clustering (AHC). The first two principal components (PC1 and PC2) explained 50.62% and 23.28% of total variance, respectively. The AHC as revealed by heat map classified cauliflower genotypes into four main groups based on measured traits. The information is useful for developing varieties and/or hybrids rich in bioactive compounds and antioxidant activity.

Dye Coupled Aptamer-Captured Enzyme Catalyzed Reaction for Detection of Pan Malaria and P. falciparum Species in Laboratory Settings and Instrument-Free Paper-Based Platform.

Malaria diagnosis methods offering species-specific information on the causative parasites, along with their flexibility to use in different resource settings, have great demand for precise treatment and management of the disease. Herein, we report the detection of pan malaria and P. falciparum species using a dye-based reaction catalyzed by the biomarker enzymes Plasmodium lactate dehydrogenase ( PLDH) and Plasmodium falciparum glutamate dehydrogenase ( PfGDH), respectively, through instrument-based and instrument-free approaches. For the detection, two ssDNA aptamers specific to the corresponding PLDH and PfGDH were used. The aptamer-captured enzymes were detected through a substrate-dependent reaction coupled with the conversion of resazurin (blue, ∼λ605nm) to resorufin (pink, ∼λ570nm) dye. The reaction was monitored by measuring the fluorescence intensity at λ660nm for resorufin, absorbance ratio (λ570nm/λ605nm), and change in color (blue to pink). The detection approach could be customized to a spectrophotometer-based method and an instrument-free device. For both the approaches, the biomarkers were captured from the serum samples with the help of aptamer-coated magnetic beads prior to the analysis to exclude potential interferences from the serum. In the instrument-free device, a medical syringe (5 mL) prefabricated with a magnet was used for in situ separation of the enzyme-captured beads from the reaction supernatant. The converted dye in the supernatant was then efficiently adsorbed over a DEAE cellulose-treated paper wick assembled in the syringe hose. The biomarkers could be detected by both qualitative and quantitative format following the color and pixel intensity, respectively, developed on the paper surface. The developed method and technique offered detection of the biomarkers within a clinically relevant dynamic range, with the limit of detection values in the picomolar level. Flexible detection capability, low cost, interference-free detections, and portable nature (for instrument-free devices) are the major advantages offered by the developed approaches.

A proteomic study of cysteine protease induced cell death in anthers of male sterile tobacco transgenic plants.

Manifestation of male sterility in plants is an important requirement for hybrid seed production. Tapetum cell layer of anther is a primary target for genetic manipulation for male sterility. In our previous report, the targeted expression of Arachis cysteine protease in tapetum led to premature degeneration of tapetal layer that resulted in complete male sterility in transgenic tobacco plants. To correlate cysteine protease mediated cell death of tapetum, transmission electron microscopy (TEM) and proteomic pattern of anthers of cysteine protease induced male sterile plant were compared with the untransformed control plant. TEM study revealed the abnormal growth of tapetal cells exhibiting excessive vacuolization that synchronized with irregular exine wall formation of the microspores. In anther proteome, a total 250 protein spots were detected that were reproducible and exhibited similar distribution pattern. Further, anther proteome of male sterile plant showed the significant upregulation (≥ 1.5) of 56 protein spots. Using Mass spectroscopy (MALDI TOF/TOF), we have identified 14 protein spots that were involved in several processes such as energy metabolism, protein synthesis, plastid protein, lipid metabolism, and cell wall assembly. Upregulation of patatin-like protein-2 homolog, carboxylesterase 17 and dicer like protein-4 in male sterile anthers that have been demonstrated to induce cell death, suggesting that cysteine protease mediated premature tapetal cell death might involve the lipid peroxidation pathway in coordination with gene silencing mechanism.

The NQO1 bioactivatable drug, ß-lapachone, alters the redox state of NQO1+ pancreatic cancer cells, causing perturbation in central carbon metabolism.

Many cancer treatments, such as those for managing recalcitrant tumors like pancreatic ductal adenocarcinoma, cause off-target toxicities in normal, healthy tissue, highlighting the need for more tumor-selective chemotherapies. ß-Lapachone is bioactivated by NAD(P)H:quinone oxidoreductase 1 (NQO1). This enzyme exhibits elevated expression in most solid cancers and therefore is a potential cancer-specific target. ß-Lapachone's therapeutic efficacy partially stems from the drug's induction of a futile NQO1-mediated redox cycle that causes high levels of superoxide and then peroxide formation, which damages DNA and causes hyperactivation of poly(ADP-ribose) polymerase, resulting in extensive NAD+/ATP depletion. However, the effects of this drug on energy metabolism due to NAD+ depletion were never described. The futile redox cycle rapidly consumes O2, rendering standard assays of Krebs cycle turnover unusable. In this study, a multimodal analysis, including metabolic imaging using hyperpolarized pyruvate, points to reduced oxidative flux due to NAD+ depletion after ß-lapachone treatment of NQO1+ human pancreatic cancer cells. NAD+-sensitive pathways, such as glycolysis, flux through lactate dehydrogenase, and the citric acid cycle (as inferred by flux through pyruvate dehydrogenase), were down-regulated by ß-lapachone treatment. Changes in flux through these pathways should generate biomarkers useful for in vivo dose responses of ß-lapachone treatment in humans, avoiding toxic side effects. Targeting the enzymes in these pathways for therapeutic treatment may have the potential to synergize with ß-lapachone treatment, creating unique NQO1-selective combinatorial therapies for specific cancers. These findings warrant future studies of intermediary metabolism in patients treated with ß-lapachone.

Transesophageal Echocardiographic Assessment of Pulmonary Artery-to-Ascending Aorta Ratio for the Detection of Pulmonary Hypertension in Cardiac Surgical Patients.

OBJECTIVE: The objective of the study was to investigate if the main pulmonary artery (mPA)-to-ascending aorta (AscAo), (mPA:AscAo) ratio could serve as a screening tool in identifying pulmonary artery hypertension (PAH). DESIGN: A prospective observational study. SETTING: Tertiary care center, university hospital. PARTICIPANTS: Fifty-four adult patients undergoing off-pump coronary artery bypass grafting surgery (OPCAB). INTERVENTIONS: mPA and AscAo transverse diameters were measured by transesophageal echocardiography (TEE) and the mean pulmonary arterial pressures (mPAP) were recorded simultaneously using a pulmonary artery catheter. MEASUREMENTS AND MAIN RESULTS: mPA:AscAo ratio demonstrated significant linear correlation with mPAP measured by pulmonary artery catheterization (ie, r = 0.61, confidence interval [CI] = 0.5352-0.6736, p < 0.0001). Receiver operating characteristic curves were performed to evaluate sensitivity and specificity of mPA:AscAo ratio ≥1 for diagnosing PAH (mPAP ≥25 mmHg). Area under the curve for mPA:AscAo ratio was 0.91 (95% CI, 0.869-0.936, p < 0.0001), with a sensitivity of 84.27%, specificity of 83.92%, positive-predictive value of 87.6% and negative-predictive value of 81.1% for a mPAP ≥25 mmHg. CONCLUSIONS: The ratio of mPA:AscAo is a simple, reliable, and reproducible method that can be obtained through TEE, which guides the clinician to screen patients with PAH.

Monophasic synovial sarcoma of posterior pharyngeal wall: a rare case report with unique reconstruction using lateral trapezius flap.

Synovial sarcoma is a rare entity to be encountered in the head and neck region and is always a challenge in terms of diagnosis, treatment planning and reconstruction of the surgical defect. In our case, we faced a similar challenge for diagnosis and also have ventured for lateral trapezius flap as a new reconstructive option for such bulky tumour defects. We hereby present a 25-year old male patient with monophasic synovial sarcoma of posterior pharyngeal wall. The radiological and clinicopathological features along with various diagnostic tests and treatment options are discussed.

Development of an Indicator Displacement Based Detection of Malaria Targeting HRP-II as Biomarker for Application in Point-of-Care Settings.

A novel label free spectrophotometric detection of malarial biomarker HRP-II following an indicator displacement assay has been developed. The assay is based on competitive displacement of murexide dye from its complex with Ni2+ by HRP-II present in serum samples. The binding constant (Kd) discerned for the dye and HRP-II to Ni2+ were 1.4 × 10-6 M-1 and 6.8 × 10-9 M-1, respectively. The progress of the reaction could be monitored from the change of color from orange (∼λ482 nm) to pink (∼λ515 nm) with the concomitant increase in HRP-II concentration in the mixture. A linear response (R2 = 0.995) curve was generated by plotting the ratio of absorbance (λ515 nm/λ482 nm) against the HRP-II concentrations. The method offers to detect HRP-II as low as 1 pM without any interference from some common salts and the major protein, HSA, present in the blood serum. The detection method was reproduced in a microfluidic paper based analytical device (µPAD), fabricated by printing hydrophobic alkyl ketene dimer on a chromatographic paper to create hydrophilic microchannels, test zone, and sample application zone. The device offers to use a maximum sample volume of 20 ± 0.06 µL and detects HRP-II within 5 min with LOD of 30 ± 9.6 nM in a dynamic range of 10 to 100 nM. The method has thus immense potential to develop as rapid, selective, simple, portable, and inexpensive malarial diagnostic device for point-of-care and low resource setting applications.

Development and validation of functional CAPS markers for the FAE genes in Brassica juncea and their use in marker-assisted selection.

Low erucic acid is a major breeding target to improve the edible oil quality in Brassica juncea. The single nucleotide polymorphism (SNP) in fatty acid elongase 1 (FAE1.1 and FAE1.2) gene was exploited to expedite the breeding program. The paralogs of FAE1 gene were sequenced from low erucic acid genotype Pusa Mustard 30 and SNPs were identified through homologous alignment with sequence downloaded from NCBI GenBank. Two SNPs in FAE1.1 at position 591 and 1265 and one in FAE1.2 at 237 were found polymorphic among low and high erucic acid genotypes. These SNPs either create or change the recognition site of restriction enzymes. Transition of a single nucleotide at position 591 and 1265 in FAE1.1, and at position 237 in FAE1.2, leads to a change in the recognition site of Hpy99I, BglII and MnlI restriction enzymes, respectively. Two CAPS markers for FAE1.1 and one for FAE1.2 were developed to differentiate low and high erucic acid genotypes. The efficiency of these CAPS markers was found 100 per cent when validated in Brassica juncea, and B. nigra genotypes and used in back-cross breeding. These CAPS markers will facilitate in marker-assisted selection for improvement of oil quality in Brassica juncea.

Molecular identification of Aedes aegypti mosquitoes from Pilani region of Rajasthan, India.

BACKGROUND & OBJECTIVES: Aedes aegypti is the most important vector of dengue virus infection in humans worldwide. Accurate identification and colonization are the essential requirements to understand vector biology as well as its diseases transmission potential. In this study, we have used molecular approaches for the identification of Ae. aegypti mosquitoes that were collected from the Pilani region of Rajasthan, India Methods: Field collected mosquito larvae were colonized under laboratory conditions. Conserved genetic markers, ITS-2 and mtCOI were used for amplification through species-specific primers to identify the mosquito species/ strain. Sequencing result of this strain was phylogenetically compared with other global strains through MEGA software. RESULTS: A comprehensive multiple sequence alignment and phylogenetic analysis revealed that COI gene of Ae. aegypti has extremely low genetic variability with one of the Indian isolate from Thirumala, Andhra Pradesh region (GenBank: HM807262.1). However, in context of different geographical locations, it indicated close similarity with Thailand's strain and high variability when compared with Madagascar strain. On the other hand, ITS-2 illustrated highest identity with Ae. aegypti of Saudi Arabia (GenBank: JX423807.1) whereas, high divergence was observed from Mayotte, France strain (GenBank: KF135506). INTERPRETATION & CONCLUSION: The findings suggest that this isolate from Rajasthan is similar to other Asian continent strains possibly due to the same origin. Understanding the vectorial capacity of these geographically distributed mosquito strains will enhance our knowledge to improve existing vector surveillance and control programme.

Characterization of a vacuolar processing enzyme expressed in Arachis diogoi in resistance responses against late leaf spot pathogen, Phaeoisariopsis personata.

Vacuolar processing enzymes are cysteine proteases responsible for maturation of vacuolar proteins. They have been shown to possess caspase-1-like activity, mediate cell death and display increased activity during pathogen infections. A transcript derived fragment corresponding to VPE was found to be up-regulated in a cDNA-AFLP analysis of host responses of a wild peanut, Arachis diogoi upon challenge from the late leaf spot pathogen Phaeoisariopsis personata, which was subsequently validated by q-PCR in a time course analysis, where susceptible peanut did not show its upregulation. In transient conditional and constitutive expression studies in tobacco leaves using agroinfiltration, we have observed that expression of AdVPE was associated with hypersensitive response (HR) like cell death. AdVPE expression was found to be high at 24 h post estradiol application and this was associated with the enhanced co-expression of molecular markers of HR cell death genes and genes for pathogenesis related proteins indicating that AdVPE positively regulates defense responses and its estradiol induced expression is sufficient for HR-like cell death in tobacco. We found that AdVPE expression was very strongly induced in response to sodium nitroprusside, which indicates its involvement in stress signaling. Induced expression of AdVPE in response to jasmonic acid and ethylene also indicates its involvement in an interconnected network of signaling. Transgenic tobacco plants ectopically expressing AdVPE exhibited enhanced resistance against Phytophthora parasitica var. nicotianae, Alternaria alternata var.  nicotianae and Rhizoctonia solani. To our knowledge, this is the first report on the heterologous expression of a pathogen induced VPE enhancing resistance to fungal pathogens with cell death phenomenon under transient expression.

Comparative study of the results of compound tibial shaft fractures treated by Ilizarov ring fixators and limb reconstruction system fixators.

PURPOSE: Ilizarov ring fixator and limb reconstruction system (LRS) fixators have been used in the management of complex tibial fractures with severe soft tissue injuries, compound tibial fractures, and infected tibial nonunion for which conventional internal fixation cannot be contemplated. Fracture union and distraction osteogenesis can be done simultaneously with these external fixators, allowing early weight bearing. Several previous studies have shown almost equal results of rail and ring fixators for the compound tibial shaft fractures. Thus we performed a prospective study to evaluate the union rate, functional outcome and amount of limb lengthening after the treatment of compound tibial shaft fractures with or without infected nonunion by ring or LRS fixators. METHODS: This prospective study was done at Sarojini Naidu Medical College and Hospital, Agra, India and included 32 patients of compound tibial shaft fractures with or without infected nonunion. There were 26 males and 6 females and the average age was 40 years. Patients were randomly divided into two groups (n=16 for each): one underwent Ilizarov fixation and the other received LRS fixation. Cases were followed up for 3-24 months, 6 months on average from September 2012 to October 2014. Functional and radiological outcomes were assessed using the Association for the Study and Application of Methods of Ilizarov (ASAMI) criteria for both rail and ring fixators. RESULTS: Union was achieved in all cases. Radiological outcome was found excellent in 68.75%, good in 18.75% and fair in 12.50% of cases treated by rail fixators whereas it was excellent in 56.25%, good in 18.75%, fair in 12.50% and poor in 12.50% of cases treated by ring fixators. Functional result was satis- factory in 75.00% of cases treated by rail fixator and 68.75% of cases treated by ring fixators whereas the corresponding rate of unsatisfactory was 25.00% vs. 31.25%. CONCLUSION: In our short-term assessment, LRS fixators show a better result than Ilizarov fixators in terms of fracture union and functional outcome with soft tissue care and limb length.