Novel report of Acinetobacter johnsonii as an indole-producing seed endophyte in Tamarindus indica L.

Plant-microbe associations have been regarded as an exciting topic of research due to their potential as environment friendly alternatives for stimulating crop growth and development. Seeds of Tamarindus indica L. have been chosen for the present study as seed endophytes prefer larger or nutritive cotyledon and hard seed coats for their colonization. The main objectives of our study were to isolate and identify the seed endophytes, their bioefficacy, and responsible chemical compounds. In a dose-dependent experiment, tamarind seed exudates (TSE) showed plant growth-promoting properties on Oryza sativa (53-81%), Daucus carota (10-31%), and Raphanus sativa (21-42%). Identification of the bacterial load in TSE through 16S rRNA sequencing revealed the existence of two bacterial species, Acinetobacter johnsonii and Niallia nealsonii. This is the first report of these two bacteria as seed endophytes of Tamarindus indica L. HRLC-MS analysis of TSE confirmed the presence of indole derivatives, primarily indole-3-lactic acid (ILA). The quantitative phytochemical estimation of bacterial culture filtrates revealed that indole-like substances were present in the extracts only in A. johnsonii at a concentration of 0.005 mg/ml of indole acetic acid equivalent. Experimental results suggested that the stimulatory activity of TSE was caused by the presence of A. johnsonii, a potential plant growth-promoting bacteria that produced indole-like compounds. This study suggests tamarind seed exudates with its endophytic microbiota as a potent plant growth-promoting agent that may find use as a cheap and sustainable source of metabolites useful in the agro-industries.

Synthesis, antimicrobial, anticancer evaluation and molecular docking with Bax and MDM2 of dibromosterculic acid.

Dibromosterculic acid [8-(1,2-dibromo-2-octylcyclopropyl)-octanoic acid], a new synthetic derivative was prepared by bromination of sterculic acid. This synthetic derivative showed strong fungicidal activity against two pathogenic fungal species namely Penicillium chrysogenum and Aspergillus niger with minimum inhibitory concentration (MIC) value of 0.007 mg/ml and good bactericidal activity against Bacillus subtilis and Xanthomonas sp. with MIC value of 0.015 mg/ml. Cytotoxic activity on both normal (MCF-10A) and cancerous (MDA-MB-468) cell lines revealed that the survivability percentage of normal cells was unaffected, whereas cancerous cells were decreased greatly by dibromosterculic acid with 50% survivability at 9 µg/ml concentration. Molecular-docking using AutoDock 4.2 with Bax exhibited strong pi-sigma interaction with PHE-93, pi-alkyl and alkyl interaction with TRP-139, ARG-89 and PHE-92 whereas MDM2 revealed strong hydrogen bond interaction with GLN-59 and pi-alkyl interaction with PHE-55. All experimental parameters suggested that this synthetic derivative would be valuable for target-specific drug development with nominal side effects.

Evolution of Organic Solvent-Resistant DNA Polymerases.

The introduction of thermostable polymerases revolutionized the polymerase chain reaction (PCR) and biotechnology. However, many GC-rich genes cannot be PCR-amplified with high efficiency in water, irrespective of temperature. Although polar organic cosolvents can enhance nucleic acid polymerization and amplification by destabilizing duplex DNA and secondary structures, nature has not selected for the evolution of solvent-tolerant polymerase enzymes. Here, we used ultrahigh-throughput droplet-based selection and deep sequencing along with computational free-energy and binding affinity calculations to evolve Taq polymerase to generate enzymes that are both stable and highly active in the presence of organic cosolvents, resulting in up to 10% solvent resistance and over 100-fold increase in stability at 97.5 °C in the presence of 1,4-butanediol, as well as tolerance to up to 10 times higher concentrations of the potent cosolvents sulfolane and 2-pyrrolidone. Using these polymerases, we successfully amplified a broad spectrum of GC-rich templates containing regions with over 90% GC content, including templates recalcitrant to amplification with existing polymerases, even in the presence of cosolvents. We also demonstrated dramatically reduced GC bias in the amplification of genes with widely varying GC content in quantitative polymerase chain reaction (qPCR). By expanding the scope of solvent systems compatible with nucleic acid polymerization, these organic solvent-resistant polymerases enable a dramatic reduction of sequence bias not achievable through thermal resistance alone, with significant implications for a wide range of applications including sequencing and synthetic biology in mixed aqueous-organic media.

Characterization and metabolomic profiling of two pigment producing fungi from infected fruits of Indian Gooseberry.

Two pigment producing fungi, Talaromyces atroroseus and Penicillium choerospondiatis, were isolated and identified from infected fruits of Phyllanthus emblica L. based on amplification and sequencing of internal transcribed spacer region and beta-tubulin gene. This is the first occurrence report of these two fungi from fruits of P. emblica. Culture extract containing metabolites of T. atroroseus and P. choerospondiatis contained phenolics of 26.35 mg and 30.89 mg GAE/g dry extract respectively; whereas no significant amount of flavonoids and tannins were detected. P. choerospondiatis metabolites extract showed higher DPPH and ABTS activity with IC50 values of 21.94 mg/ml and 27.03 mg/ml respectively than T. atroroseus. LC-HRMS analysis of metabolites extract of T. atroroseus revealed presence of trimethyl-isopropyl-butanamide, perlolyrine, N-hexadecanoylpyrrolidine etc. whereas P. choerospondiatis displayed presence of tangeraxanthin, ugaxanthone, daphniphylline, etc. Therefore, fungal metabolites are rich natural sources of diversified compounds that can be utilized in dyeing industries, cosmetics and novel drug development.

Photonic Nano-/Microstructured Diatom Based Biosilica in Metal Modification and Removal-A Review.

The siliceous exoskeletal shells of diatoms, commonly known as frustules, have drawn attention because of their photoluminescence property and high volume to surface area. Photonic biosilica can also enhance the plasmonic sensitivity of nanoparticles. Because of this, researchers have studied the effectiveness of various metal particles after combining with biosilica. Additionally, naturally occurring diatom-based biosilica has excellent adsorption and absorption capabilities, which have already been exploited for wastewater treatment. Moreover, the nanoporous, ultra-hydrophilic frustules can easily accumulate more molecules on their surfaces. As a consequence, it becomes easier to conjugate noble metals with silica, making them more stable and effective. The main focus of this review is to agglomerate the utility of biocompatible diatom frustules, which is a no-cost natural resource of biosilica, in metal modification and removal.

The rolB-transgenic Nicotiana tabacum plants exhibit upregulated ARF7 and ARF19 gene expression.

Agrobacterium rhizogenes root oncogenic locus B (rolB) is known to induce hairy roots along with triggering several physiological and morphological changes when present as a transgene. However, it is still unknown how this gene triggers these changes within the plant system. In this study, the effect of rolB in-planta, when present as a transgene, was assessed on the gene expression levels of auxin response factors (ARFs)-transcription factors which are key players in auxin-mediated responses. The goal was to uncover Auxin/ARF-driven transcriptional networks potentially active and working selectively, if any, in rolB transgenic background, which might potentially be associated with hairy root development. Hence, the approach involved establishing rolB-transgenic Nicotiana tabacum plants, selecting ARFs (NtARFs) for context-relevance using bioinformatics followed by gene expression profiling. It was observed that out of the chosen NtARFs, NtARF7 and NtARF19 exhibited a consistent pattern of gene upregulation across organ types. In order to understand the significance of these selective gene upregulation, ontology-based transcriptional network maps of the differentially and nondifferentially expressed ARFs were constructed, guided by co-expression databases. The network maps suggested that NtARF7-NtARF19 might have major deterministic, underappreciated roles to play in root development in a rolB-transgenic background-as observed by higher number of "root-related" biological processes present as nodes compared to network maps for similarly constructed other non-differentially expressed ARFs. Based on the inferences drawn, it is hypothesized that rolB, when present as a transgene, might drive hairy root development by selective induction of NtARF7 and NtARF19, suggesting a functional link between the two, leading to the specialized and characteristic rolB-associated traits.

Present status and future directions: vertical root fractures in root filled teeth.

Vertical root fracture (VRF) is a common reason for the extraction of root filled teeth. The accurate diagnosis of VRF may be challenging due to the absence of clinical signs, whilst conventional radiographic assessment is often inconclusive. However, an understanding of the aetiology of VRFs, and more importantly, the key predisposing factors, is crucial in identifying teeth that may be susceptible. Thorough clinical examination with magnification and co-axial lighting is essential in identifying VRFs, and although CBCT is unable to reliably detect VRFs per se, the pattern of bone loss typically associated with VRF can be fully appreciated, and therefore, increases the probability of correct diagnosis and management. The prevalence of VRFs in root filled teeth is significantly greater than in teeth with vital pulps, demonstrating that the combination of loss of structural integrity, presence of pre-existing fractures and biochemical effects of loss of vitality is highly relevant. Careful assessment of the occlusal scheme, presence of deflective contacts and identification of parafunctional habits are imperative in both preventing and managing VRFs. Furthermore, anatomical factors such as root canal morphology may predispose certain teeth to VRF. The influence of access cavity design and root canal instrumentation protocols should be considered although the impact of these on the fracture resistance of root filled teeth is not clearly validated. The post-endodontic restoration of root filled teeth should be expedient and considerate to the residual tooth structure. Posts should be placed 'passively' and excessive 'post-space' preparation should be avoided. This narrative review aims to present the aetiology, potential predisposing factors, histopathology, diagnosis and management of VRF and present perspectives for future research. Currently, there are limited options other than extraction for the management of VRF, although root resection may be considered in multi-rooted teeth. Innovative techniques to 'repair' VRFs using both orthograde and surgical approaches require further research and validation. The prevention of VRFs is critical; identifying susceptible teeth, utilizing conservative endodontic procedures, together with expedient and appropriate post-endodontic restorative procedures is paramount to reducing the incidence of terminal VRFs.

Spatiotemporal variations in the composition of algal mats in wastewater treatment ponds of tannery industry.

Wastewater Treatment Pond (WTP) is an effective remediation technology for economically developing nations. Although it's excessive organic and nutrient loads with higher water logging time triggers mixed and unprofitable microalgal mats. This may serve as a seeding source for Cyanobacterial bloom in receiving waterbodies. Since, to maintain the growth of desirable algal species in WTPs, understanding towards environmental regulation and algal mat composition is important, especially for tropical countries, like India. In this study, biological treatment pond (BTP) and outlet pond (OP), of a tannery effluent treatment plant in eastern coast of India, were chosen for surveying the algal community composition concerning ecological parameters. Nearly, both the ponds were polluted, but the diversity was lower in BTP due to its elevated nutrient content (Ammonia 173 mg L-1) and higher persistent organic matters (COD 301.7 mg L-1) than OP. Using canonical correspondence analysis, seasonal variations showed higher species abundance during early summer compared to other seasons. A total of 37 taxa forming thick algal mats were recorded. The matrix of mats was mainly composed of Cyanobacterial members such as Phormidium, Leptolyngbya, Spirulina, and Pseudanabaena, followed by diatoms, especially Amphora and Nitzschia. Diatoms commonly occurred as embedded component in the entangled matrix of blue-green algal filaments. Hierarchical cluster analysis was employed to group all these taxa based on their seasonal appearance and abundance. This year-long intensive study revealing seasonal algal mat composition patterns in these WTPs will ultimately safeguard the livelihood and security of adjoining localities through proper site-specific pollution control.

In-situ green synthesis of fluorescent silica-silver conjugate nanodendrites using nanoporous frustules of diatoms: an unprecedented approach.

Generally, nanodendrite synthesis is chemical mediated and expensive. The biogenesis of such hierarchical structures is still in its nascent stage. The present study aimed at exploiting the nanoporous frustules of Halamphora subturgida, as a source of biosilica for the biosynthesis and stabilization of conjugate nanodendrites of silica and silver. These minute diatom frustules when exposed to 9 mM of silver nitrate solution, a highly crystalline nanohybride dendrites were synthesized. The nanohybrid dendrite synthesis was initially confirmed by the formation of greyish-brown frustules after 72 h of exposure. The composite dendrites were thoroughly characterized by standard techniques. Electron microscopic images illustrated that the process began with the formation of isotropic hybrid nanospheres with an internal diameter of 20 nm and continued to develop anisotropic nanocrystals with time. The nanodendrites externally formed on the siliceous frustules, acting as a template for the former. They were characterized by distinct 100 nm wide and 1-2 µm long trunks and 70-100 nm wide and 220-220 nm long branches on either side of the trunk. The optical measurement revealed the fluorescence property of the nanostructures owing to the photoluminescent efficiency of the frustules. Both the externally derived hybrid nanodendrites and internally synthesized nanospheres possessed superior stability in the suspension with a zeta potential value of - 35.7 mV and - 24.8 mV, respectively. Thus, this method is eco-friendly and provides a new dimension for nanodendrite synthesis with minimal cost and maximal yield compared to its non-biologically synthesized counterparts that involve several other drawbacks like chemical hazards and high energy consumption.

Antimicrobial Effectiveness of Calcium Silicate Sealers against a Nutrient-Stressed Multispecies Biofilm.

PURPOSE: This study compared the antimicrobial efficacy of calcium silicate sealers (BioRoot RCS and Total Fill BC) and conventional sealers (AH Plus and Tubli-seal) against planktonic bacteria and a nutrient-stressed multispecies biofilm. METHODS: Antimicrobial properties of freshly mixed sealers were investigated using the direct contact test (DCT) and a nutrient-stressed multispecies biofilm comprised of five endodontic strains. Antimicrobial activity was determined using quantitative viable counts and confocal laser scanning microscopy (CLSM) analysis with live/dead staining. The pH of the sealers was analysed over a period of 28 days in Hanks Balanced Salt Solution (HBSS). Analysis of variance (ANOVA) with Tukey tests and the Kruskal-Wallis test were used for data analysis with a significance of 5%. RESULTS: All endodontic sealers exhibited significant antimicrobial activity against planktonic bacteria (p < 0.05). BioRoot RCS caused a significant reduction in viable counts of the biofilms compared to AH Plus and the control (p < 0.05), while no significant difference could be observed compared to TotalFill BC and Tubli-seal (p > 0.05). CLSM analysis showed that BioRoot RCS and TotalFill BC exhibited significant biofilm inhibition compared to Tubli-seal, AH Plus and the control (p < 0.05). BioRoot RCS presented with the highest microbial killing, followed by TotalFill BC and Tubli-seal. Alkalizing activity was seen from the onset by BioRoot RCS, TotalFill BC and AH Plus. After 28 days, BioRoot RCS demonstrated the highest pH in HBSS (pH > 12). CONCLUSIONS: Calcium silicate sealers exhibited effective antimicrobial properties. This was demonstrated by superior biofilm inhibition capacity and microbial killing, with strong alkalizing activity compared to epoxy-based and zinc oxide-eugenol-based sealers.

Enhancement of ABA Sensitivity Through Conditional Expression of the ARF10 Gene in Brassica juncea Reveals Fertile Plants with Tolerance Against Alternaria brassicicola.

Concomitant increase of auxin-responsive factors ARF16 and ARF17, along with enhanced expression of ARF10 in resistant Sinapis alba compared with that in susceptible Brassica juncea upon challenge with Alternaria brassicicola, revealed that abscisic acid (ABA)-auxin crosstalk is a critical factor for resistance response. Here, we induced the ABA response through conditional expression of ARF10 in B. juncea using the A. brassicicola-inducible GH3.3 promoter. Induced ABA sensitivity caused by conditional expression of ARF10 in transgenic B. juncea resulted in tolerance against A. brassicicola and led to enhanced expression of several ABA-responsive genes without affecting the auxin biosynthetic gene expression. Compared with ABI3 and ABI4, ABI5 showed maximum upregulation in the most tolerant transgenic lines upon pathogen challenge. Moreover, elevated expression of ARF10 by different means revealed a direct correlation between ARF10 expression and the induction of ABI5 protein in B. juncea. Through in vitro DNA-protein experiments and chromosome immunoprecipitation using the ARF10 antibody, we demonstrated that ARF10 interacts with the auxin-responsive elements of the ABI5 promoter. This suggests that ARF10 may function as a modulator of ABI5 to induce ABA sensitivity and mediate the resistance response against A. brassicicola.

A critical review on use of Agrobacterium rhizogenes and their associated binary vectors for plant transformation.

Agrobacterium rhizogenes, along with A. tumefaciens, has been used to affect genetic transformation in plants for many years. Detailed studies conducted in the past have uncovered the basic mechanism of foreign gene transfer and the implication of Ri/Ti plasmids in this process. A number of reviews exist describing the usage of binary vectors with A. tumefaciens, but no comprehensive account of the numerous binary vectors employed with A. rhizogenes and their successful applications has been published till date. In this review, we recollect a brief history of development of Ri-plasmid/Ri-T-DNA based binary vectors systems and their successful implementation with A. rhizogenes for different applications. The modification of native Ri plasmid to introduce foreign genes followed by development of binary vector using Ri plasmid and how it facilitated rapid and feasible genetic manipulation, earlier impossible with native Ri plasmid, have been discussed. An important milestone was the development of inducible plant expressing promoter systems which made expression of toxic genes in plant systems possible. The successful application of binary vectors in conjunction with A. rhizogenes in gene silencing and genome editing studies which are relatively newer developments, demonstrating the amenability and adaptability of hairy roots systems to make possible studying previously intractable research areas have been summarized in the present review.

Isolated congenitally corrected transposition of the great arteries with dextroversion discovered incidentally in a patient with cocaine-induced acute myocardial infarction.

Complex cardiac congenital anomalies can occasionally be found in adult patients who have no knowledge of their condition. Here we present the case of a 27-year-old man with cocaine-induced acute myocardial infarction in whom an isolated congenitally corrected transposition of the great arteries with dextroversion was discovered incidentally.

Genetic and biochemical consequences of adenosine deaminase deficiency in humans.

Adenosine deaminase deficiency accounts for approximately 15-20% of severe combined immunodeficiency in humans. The gene for adenosine deaminase is located on chromosome 20q12-q13.11 and codes for an aminohydrolase that catalyzes the deamination of adenosine and deoxyadenosine to inosine and deoxyinosine, respectively. Absence of the enzyme causes a build-up of the substrates in addition to excess deoxyadenosine triphosphate, thereby compromising the regenerative capacity of the immune system. Due to underlying allelic heterogeneity, the disorder manifests as a spectrum, ranging from neonatal onset severe combined immunodeficiency to apparently normal partial adenosine deaminase deficiency. Tandem mass spectrometry coupled with high efficiency separation systems enables postnatal diagnosis of the disorder, while prenatal diagnosis relies on assaying enzyme activity in cultured amniotic fibroblasts or chorionic villi sampling. Screening of adenosine deaminase deficiency for relatives-at-risk may reduce costs of treatment and ensure timely medical intervention as applicable. This article reviews the genetic, biochemical and clinical aspects of adenosine deaminase deficiency.

Use of Angio-Seal closure device when the arteriotomy is above or below the common femoral artery.

BACKGROUND: The majority of vascular complications during diagnostic coronary angiography occur while accessing the femoral artery. Angio-Seal vascular closure devices are indicated for closure of arteriotomies of the femoral artery, but not when the puncture site is above or below the femoral artery. HYPOTHESIS: Under certain circumstances, use of the Angio-Seal closure device above or below the common femoral artery may be safe and effective. METHODS: Consecutive patients at a single high-volume medical center having documented high or low arteriotomies were closed using an Angio-Seal closure device. Patients were chosen when they received a single arterial puncture, the arteriotomy was away from branch vessels, and the diameter of the artery at the site of sheath entry was of large enough caliber to suggest placement would be successful. Patients were then followed longitudinally for complications. RESULTS: Sixty-two patients had non-femoral artery sheath placement. Twenty-two patients had "high" puncture sites, and 40 patients had "low" puncture sites. All were successfully closed using the Angio-Seal vascular closure device and had no complications. CONCLUSIONS: Although not approved for placement in areas above or below the femoral artery, off-label use of the Angio-Seal arteriotomy closure may be safely performed when patient characteristics are favorable.

The AmBulatory Closure Device Percutaneous Intervention (ABCD-PCI) study: a single-center experience.

The AmBulatory Closure Device Percutaneous Intervention (ABCD-PCI) study is a multicenter randomized prospective controlled trial evaluating the safety of and patient satisfaction with same-day discharge following ambulatory percutaneous intervention with a closure device. This article reviews the findings from a single center, Baylor Jack and Jane Hamilton Heart and Vascular Hospital, from October 2008 through April 2010, with 23 patients in the same-day discharge group and 21 patients in the next-day discharge group. There were no differences between the groups in demographic or procedure characteristics. Outcomes were measured by a questionnaire and 7-day and 30-day follow-up phone calls. Results showed that same-day discharge after percutaneous intervention with a closure device is as safe as next-day discharge. However, there was a trend for a higher comfort level among patients in the next-day discharge group.

Successful percutaneous retrieval of methyl methacrylate orthopedic cement embolism from the pulmonary artery.

Vertebroplasty cement embolization into the venous system has long been recognized as a potential complication, but the true incidence of systemic embolization is unknown. Clinical presentations range from patients who are asymptomatic or have incidental findings on imaging to massive pulmonary embolism resulting in death. Optimal treatment is controversial and the natural history is unknown. We present the case of an 85-year-old female undergoing combined laminectomy and vertebroplasty with subsequent pulmonary embolism of the cement which was successfully retrieved from a percutaneous approach.

Identifying cardiovascular syphilis at operation.

To describe the morphologic features so the process can be easily identified during surgery, we studied 34 patients with cardiovascular syphilis, 32 of whom underwent excision and replacement of the ascending aorta or aortic valve or both. Of the 34 patients, 22 were treated at Baylor University Medical Center from 1998 to 2008 and 12 at non-Baylor University Medical Center hospitals from 1958 to 1987. In all 34 patients, the tubular portion of the aorta was diffusely thickened and the sinus portion of the aorta was apparently uninvolved. The process involved all 3 layers of the aorta, with thickening of the adventitia, mainly by fibrous tissue. Within the fibrous tissue were collections of plasma cells and lymphocytes, focal destruction of the media without thickening, and marked thickening of the intima by atherosclerotic-appearing lesions. Serologic tests for syphilis were done in only 14 patients (41%) and were positive (reactive) in 6 (43%) of them. The ascending aorta, however, was similar in all 34 patients. In conclusion, cardiovascular syphilis has not disappeared. Its identification during surgery can prompt appropriate antibiotic therapy postoperatively. Although the serologic test results for syphilis might be negative, antibiotic therapy is recommended for patients with panaortitis requiring resection of the ascending aorta with or without aortic regurgitation.

Luminescent Solar Concentrators--a review of recent results.

Luminescent solar concentrators (LSCs) generally consist of transparent polymer sheets doped with luminescent species. Incident sunlight is absorbed by the luminescent species and emitted with high quantum efficiency, such that emitted light is trapped in the sheet and travels to the edges where it can be collected by solar cells. LSCs offer potentially lower cost per Wp. This paper reviews results mainly obtained within the framework of the Full-spectrum project. Two modeling approaches are presented, i.e., a thermodynamic and a ray-trace one, as well as experimental results, with a focus on LSC stability.

Safety of drug-eluting stents in the coronary artery in ST-elevation myocardial infarction at a single high-volume medical center.

Several trials have shown the effectiveness of drug-eluting stents (DES) in reducing restenosis. Acute ST-elevation myocardial infarction (STEMI) has been an exclusion criterion in most trials evaluating the safety and efficacy of DES. There is recent randomized trial data evaluating the use and safety of DES for acute myocardial infarction. However, there is a need for "real world" data on the efficacy and safety of DES in STEMI. A single-center retrospective analysis was performed on 188 consecutive patients with STEMI treated with primary or rescue coronary angioplasty between March 2004 and July 2005. The study consisted of 3 groups: 115 patients treated with paclitaxel-eluting stents, 55 with sirolimus-eluting stents, and 18 with bare metal stents. Outcomes were assessed from 12 to 28 months (mean 20, median 19) for major adverse cardiac events (MACEs) including myocardial infarction, in-stent thrombosis, clinical restenosis, and death. There were 4 in-stent thromboses in the paclitaxel group (3.4%) and 2 in-stent thromboses in the sirolimus group (3.6%). The thromboses ranged from acute (within 24 hours) to as late as 8 months. Clinical restenosis occurred in 4 patients (3.4%) in the paclitaxel group and in 2 patients (3.6%) in the sirolimus. None of the 18 patients with bare metal stents had thrombosis or clinical restenosis. There were 7 total deaths, all related to complications from the index STEMI: 1 in the bare metal group, 1 in the sirolimus group, and 5 in the paclitaxel group. The postdischarge MACE rate was 7% with no deaths. In conclusion, the use of DES in acute STEMI is associated with a low postdischarge MACE rate and a 3.5% in-stent thrombosis rate, which is similar to reported rates in earlier randomized trials.