A Versatile Protocol for Efficient Transformation and Regeneration in Mega Indica Rice Cultivar MTU1010: Optimization through Hormonal Variables.

Rice is one of the apex food crops in terms of meeting the daily calorific and dietary requirement of the majority of the world population. However, rice productivity is severely limited by various biotic and abiotic attributes, causing a severe threat to global food security. In the use of functional genomics and genome editing for the generation of trait-enhanced genotypes, it is necessary to have an efficient genetic transformation and regeneration protocol. The recalcitrant nature and paucity of efficient and versatile genetic transformation and regeneration protocols for indica cultivars remains a constraint. In the present study, we have optimized a tissue culture method for MTU1010, a mega indica rice variety. We conducted a combinatorial analysis of different plant growth regulators on embryogenic callus induction efficiency, and it was observed that MSB5 medium supplemented with 2.5 mg/L 2-4D and 0.25 mg/L 6-BAP results in maximum embryogenic callus induction, i.e., 92%. The regeneration efficiency of a transformed callus can be enhanced by up to 50% with the supplementation of 1 mg/L kinetin alongside 2.5 mg/L BAP and 0.5 mg/L NAA in the shooting medium. Furthermore, our results unveiled that the pre-activation of Agrobacterium culture for 30 min with 150 µM acetosyringone significantly increased the transformation efficiency of calli. Additionally, descaling the salt concentration to half strength in resuspension and co-cultivation increased the efficiency of transformation up to 33%. Thus, the protocol developed in this study will be instrumental for the genome editing and genetic engineering of indica rice cultivars for functional genomics studies and crop improvement.

Optimization of Oral Posaconazole Step Down Therapy in Management of Rhino-Orbital-Cerebral Mucormycosis (ROCM): Outcome of an Institutional Protocol.

All cases of ROCM received antifungal for a prolong period in step down phase. There is no definite guidelines regarding duration. We have formulated and followed an institutional protocol with good results. To determine the outcome of an institutional protocol and required optimum duration of oral posaconazole therapy to cure ROCM in step down phase, 30 ROCM patients were followed up in step down phase. Oral posaconazole tablet was given for minimum 3, 4 & 1/2 and 6 months to stage-II, III & IV cases respectively, with a provision of extension for another 6 weeks in suspected active diseases. Nasal endoscopy, MRI, histopathology /culture of suspected tissue and hematologic investigations were done routinely to identify the residual active disease or recurrences at earliest. Outcome of this protocol proved excellent as all cases of ROCM were cured. Oral posaconazole step down therapy for a duration according to the stages of disease cured 18 cases (60%) of ROCM. Fourteen patients required extended duration of therapy as per provision of our treatment protocol. Only one patient needed re-debridement. The cases with deformities needed extended period of posaconazole therapy. No one needed posaconazole therapy for more than 7and1/2 months.

Rapid microwave hydrothermal processed spinel Co3O4nanospheres infused N-doped graphene nanosheets for high-performance battery.

Spinel Co3O4nanospheres have been synthesized by the microwave-assisted hydrothermal method. The N-doped graphene nanosheets (NGN) were synthesized using Hummer's method. The prepared spinel Co3O4and NGN were mixed under certain proportions using an ultrasonication process and treated with microwave radiation to prepare a novel spinel Co3O4nanospheres infused NGN. The synthesized samples were characterized by x-ray diffraction, Raman spectroscopy, Zetasizer, scanning electron microscope/transmission electron microscopy and x-ray photoelectron spectroscopy for identifying crystal structure and phase, particle size, and the morphology of the nanostructure and the elemental configuration, respectively. The prepared spinel Co3O4/NGN were used as anode material and lithium metal as a reference electrode to fabricate half cell using Swagelok cell components. The electrochemical properties were studied and found to exhibit a larger specific capacity of 575 mAh g-1compared to traditional graphite electrodes, after 100 cycles under 0.1 C rate with a coulombic efficiency of ≈100%. The good electrochemical properties ascribe to the distinctive surface morphological nanostructures of nanoporous nanospheres of spinel Co3O4nanospheres and nanosheets of N-doped graphene that reduce the lithium-ion diffusion pathway. The developed anode material would be a potential electrode for lithium ion battery applications.

Supercontinuum generation from zinc borate glasses: bandgap versus rare-earth doping.

We report an enhanced supercontinuum generation (SCG) from a rare-earth (Pr3+) doped low bandgap zinc borate glass when excited with 60 fs pulses from a Ti-sapphire laser. The emission associated with the absorption bands due to Pr3+ doping (around 420-500 and 580-600 nm) is observed to assist the enhanced SCG.

The abscisic acid receptor OsPYL6 confers drought tolerance to indica rice through dehydration avoidance and tolerance mechanisms.

Abscisic acid (ABA) is a key regulator of plant development and stress tolerance. Here we report functional validation of the ABA receptor OsPYL6 by constitutive and stress-inducible overexpression and RNAi silencing, in an indica rice cultivar 'Pusa Sugandh 2'. Overexpression of OsPYL6 conferred ABA hypersensitivity during germination and promoted total root length. Overexpression and RNAi silencing of OsPYL6 resulted in enhanced accumulation of ABA in seedlings under non-stress conditions, at least, in part through up-regulation of different 9-cis epoxycarotenoid dioxygenase (NCED )genes. This suggests that PYL6 expression is crucial for ABA homeostasis. Analysis of drought tolerance of OsPYL6 transgenic and wild type plants showed that OsPYL6 overexpression enhanced the expression of stress-responsive genes and dehydration tolerance. Transgenic rice plants overexpressing OsPYL6 with AtRD29A (Arabidopsis thaliana Responsive to Dehydration 29A) promoter also exhibited about 25% less whole plant transpiration, compared with wild type plants under drought, confirming its role in activation of dehydration avoidance mechanisms. However, overexpression of PYL6 reduced grain yield under non-stress conditions due to reduction in height, biomass, panicle branching and spikelet fertility. RNAi silencing of OsPYL6 also reduced grain yield under drought. These results showed that rice OsPYL6 is a key regulator of plant development and drought tolerance, and fine-tuning of its expression is critical for improving yield and stress tolerance.

CRISPR-Cas9 mediated genome editing of drought and salt tolerance (OsDST) gene in indica mega rice cultivar MTU1010.

Development of abiotic stress tolerant rice cultivars is necessary for sustainable rice production under the scenario of global climate change, dwindling fresh water resources and increase in salt affected areas. Several genes from rice have been functionally validated by using EMS mutants and transgenics. Often, many of these desirable alleles are not available indica rice which is mainly cultivated, and where available, introgression of these alleles into elite cultivars is a time and labour intensive process, in addition to the potential introgression of non-desirable genes due to linkage. CRISPR-Cas technology helps development of elite cultivars with desirable alleles by precision gene editing. Hence, this study was carried out to create mutant alleles of drought and salt tolerance (DST) gene by using CRISPR-Cas9 gene editing in indica rice cv. MTU1010. We used two different gRNAs to target regions of DST protein that might be involved in protein-protein interaction and successfully generated different mutant alleles of DST gene. We selected homozygous dst mutant with 366 bp deletion between the two gRNAs for phenotypic analysis. This 366 bp deletion led to the deletion of amino acid residues from 184 to 305 in frame, and hence the mutant was named as dst ∆184-305 . The dst ∆184-305 mutation induced by CRISPR-Cas9 method in DST gene in indica rice cv. MTU1010 phenocopied EMS-induced dst (N69D) mutation reported earlier in japonica cultivar. The dst ∆184-305 mutant produced leaves with broader width and reduced stomatal density, and thus enhanced leaf water retention under dehydration stress. Our study showed that the reduction in stomatal density in loss of function mutants of dst is, at least, in part due to downregulation of stomatal developmental genes SPCH1, MUTE and ICE1. The Cas9-free dst ∆184-305 mutant exhibited moderate level tolerance to osmotic stress and high level of salt stress in seedling stage. Thus, dst mutant alleles generated in this study will be useful for improving drought and salt tolerance and grain yield in indica rice cultivars.

Po uptake in microalgae at different seawater pH: An experimental study simulating ocean acidification.

Climate change effects such as ocean acidification (OA) are known to affect the trace metal distribution. This experimental study provides the first data on 209Po uptake rates and 210Po concentration in five microalgae species under different pH scenarios. The experiment was conducted in replicates at three pH conditions 8.2, 8.0, and 7.5, representing the current and future climate change scenario as per IPCC RCP8.5. The 209Po uptake in the phytoplankton was highest in Thalassiosira weissflogi, i.e. 83% of the 209Po tracer was taken up at 8.2 pH whereas the lowest uptake was observed in Dunaliella salina equivalent to 20% at 7.5 pH. Similar behavior was observed in 210Po concentrations in these microalgae, where 210Po ranged between 3.16 ± 0.03 and 11.6 ± 0.04 Bq kg-1 wet weight (ww), with the highest in the Thalassioria weissflogi at 8.2 pH, and the lowest in Dunaliella salina at 7.5 pH. The difference in 209Po uptake and 210Po concentration was statistically significant (p < 0.001) both among species and the pH treatments in the order: Thalassiosira weissflogi > Tetraselmis suecica > Chaetoceros muelleri > Isochrysis galbana > Dunaliella salina and 8.2 > 8.0 > 7.5. A higher concentration of 209Po in seawater was measured at low pH condition in all the experimental tanks. Though the data clearly show the difference in concentration and uptake of polonium at different pH conditions, it is not known if lower pH is affecting the adsorbed or absorbed fraction. A detailed investigation will be required to understand the process as it can have a significant effect on biomagnification and marine food chain transfer under changing climatic scenarios.

Osseous response on linear and cyclic RGD-peptides immobilized on titanium surfaces in vitro and in vivo.

Biomimetic surface modifications of titanium (Ti) implants using the Arg-Gly-Asp-sequence (RGD) are promising to accelerate bone healing in cases of medical implants. Therefore, we compared the impact of linear and cyclic RGD (l- and c-RGD) covalently coupled onto Ti surfaces on the osseous response in vitro and in vivo. In vitro, osteoblasts' behavior on different surfaces (unmodified, amino-silanized [APTES], l- and c-RGD) was analysed regarding adhesion (fluorescence microscopy), proliferation (resazurin stain) and differentiation (reverse transcription polymerase chain reaction on alkaline phosphatase and osteocalcin). In vivo, osteosynthesis screws (unmodified n = 8, l-RGD n = 8, c-RGD n = 8) were inserted into the proximal tibiae of 12 rabbits and evaluated for bone growth parameters (bone implant contact [%] and vertical bone apposition [VBA;%]) at 3 and 6 weeks. In vitro, c- as well as l-RGD surfaces stimulated osteoblasts' adherence, proliferation and differentiation in a similar manner, with only subtle evidence of superiority of the c-RGD modifications. In vivo, c-RGD-modifications led to a significantly increased VBA after 3 and 6 weeks. Thus, coating with c-RGD appears to play an important role influencing osteoblasts' behaviour in vitro but especially in vivo. These findings can be applied prospectively to implantable biomaterials with hypothetically improved survival and success rates. © 2017 Wiley Periodicals Inc. J Biomed Mater Res Part A: 106A: 419-427, 2018.

Recyclable cross-linked laccase aggregates coupled to magnetic silica microbeads for elimination of pharmaceuticals from municipal wastewater.

In the present work, the use of magnetic mesoporous silica microbeads (MMSMB) as supports was proposed to produce magnetically-separable cross-linked enzyme aggregates (MCLEAs). The effects of cross linking time, addition of bovine serum albumin as protein feeder, pH, glutaraldehyde concentration, and laccase:MMSMB mass ratio on the immobilization yield and enzyme load were investigated. The best conditions allowed the rapid preparation of MCLEAs with high enzyme load, i.e., 1.53 U laccase/mg MCLEAs. The stability of MCLEAs was improved with regard to low pH, presence of chemical denaturants, and real wastewater matrix, compared to free laccase. In addition, the novel biocatalyst exhibited good operational stability, maintaining up to 70 % of its initial activity after 10 successive batch reactions. Finally, MCLEAs demonstrated its catalytic potential to transform acetaminophen and various non-phenolic pharmaceutical active compounds as mefenamic acid, fenofibrate, and indomethacin from biologically treated wastewater effluent, with similar or even higher efficiency than free laccase.

Prospective study to evaluate safety and efficacy of Zotarolimus Eluting Stent (PSEZES) in patients with long coronary artery lesions.

BACKGROUND: Although the safety and effectiveness of Drug-Eluting Stents (DES) has been established extensively, reports on long term clinical outcome with angiographic findings in patients with long coronary artery lesions are not many. METHODS: In this single-center prospective registry of 100 patients, a total of 110 denovo long lesions (>20 mm) were treated with Resolute Zotarolimus Eluting Stent (R-ZES). The patients were followed up clinically at 3, 6 and 12 months and follow up coronary angiography was performed at 9-months. The primary end point was one year rate of target lesion failure (TLF) which is a composite of cardiac death, target lesion myocardial infarction or ischemia driven target lesion revascularization (TLR). The secondary end points included definite or probable stent thrombosis, 9-month angiographic restenosis and late lumen loss. RESULTS: The mean age of patients was 58.7 ± 9.50 years with prevalence of diabetes as high as 60%. The mean lesion length was 24.67 ± 4.87 mm with a mean reference vessel diameter of 2.85 ± 0.32 mm and 67.3% were Type C lesions (ACC/AHA classification). Two patients died during follow-up, of which one was non-cardiac death. One patient had target vessel myocardial infarction and five patients (4.5%) had ischemia driven TLR. The incidence of TLF was 6.36%. Binary restenosis was seen in 7 out of 93 lesions (7.5%). Median late lumen loss at 9 month was 0.22 mm. No stent thrombosis was noted in the study. CONCLUSION: Implantation of R-ZES in real-world patients with long coronary artery lesions is safe with comparable efficacy to what is observed in the treatment of less complex lesions.

Optical properties of Nd3+ doped bismuth zinc borate glasses.

Glasses with compositions of (100-x) (Bi2ZnOB2O6) -x Nd2O3 (where x=0.1, 0.3, 0.5, 1 and 2 mol%) were prepared by melt quenching method and characterized through optical absorption, emission and decay curve measurements. Optical absorption spectra have been analyzed using Judd-Ofelt theory. The emission spectra exhibit three peaks at 919, 1063 and 1337 nm corresponding to (4)F3/2 to (4)I9/2, (4)I11/2 and (4)I13/2 transitions in the near infrared region. The emission intensity of the (4)F3/2 to (4)I11/2 transition increases with increase of Nd(3+) concentration up to 1 mol% and then concentration quenching is observed for 2 mol% of Nd(3+) concentration. The lifetimes for the (4)F3/2 level are found to decrease with increase in Nd2O3 concentration in the glasses. The decay curves of the glass up to 0.3 mol% of Nd(3+) exhibit single exponential nature and thereafter the curves become nonexponential nature (0.5, 1 and 2 mol%). The nonexponential curve has been fitted to the Inokuti-Hirayama model to understand the nature of energy transfer process.

Associations between single-nucleotide polymorphisms of the VEGF gene and long-term prognosis of oral squamous cell carcinoma.

INTRODUCTION: Functional polymorphisms (SNPs) of the vascular endothelial growth factor (VEGF) are associated with the incidence of oral squamous cell carcinoma (OSCC). An impact of VEGF-SNPs on prognosis of OSCC patients seems possible. Therefore, correlations between prognostic parameters of OSCC patients and five VEGF-SNPs were determined. MATERIALS AND METHODS: In a retrospective long-term study, in 113 OSCC patients that underwent curative resections, five VEGF-SNPs (-1154 G/A, +405 G/C, +936 C/T, -2578 C/A, and -460 C/T) were analyzed. Associations between SNPs and prognosis (incidence of local recurrent disease, second cancer, metastases, death, total disease-free survival) were examined. RESULTS: After a mean follow-up time of 57.6 months, 32 patients had local recurrences; 15 patients had second cancer, 15 patients metastases, and 23 patients died. The mean disease-free survival was 43.1 months. A significant increased incidence of OSCC in smokers with the VEGF -2578 A/C and -460 C/T SNP was seen (each P < 0.0001). In univariate analysis, patients with advanced OSCCs (T > 2 or N > 0) together with the -1154 A/A allele had a significant worse survival and a worse disease-free survival (both P < 0.04). The same was seen for the +405 G/G SNP (both P = 0.002). In multivariate analysis, only the negative influence of the +405 G/G SNP on survival in advanced OSCCs (T > 2) could be confirmed (P = 0.002). DISCUSSION: Possible reciprocal interactions between smoking and VEGF-SNP function were observed. Multivariate analysis confirmed the VEGF +405 G/G genotype to be associated with poor survival in advanced OSCCs; a further use of this haplotype as biomarker has to be discussed.

Evaluation of subgingival microflora in diabetic and nondiabetic patients.

The present investigation was designed with the aim of studying the microbiota of diabetic patients-both insulin dependent and noninsulin dependent and nondiabetic individuals. Each of the three groups had 15 patients, coming under the age group of 35 to 55 years and all having periodontitis. Even though the microbial flora are almost the same, specific microorganisms may not be predisposing cause for the periodontal disease process in diabetics. The study is clinically significant by means of its implication in the treatment of bacterial infections related to periodontitis and in those patients who are having systemic diseases, like diabetes along with poor periodontal condition and infections.

Recording 'sublingual crescents' in lower complete dentures: a technique so effective but still esoteric and arcane.

AIM: To achieve retention in severely resorbed mandibular ridges during impression making and to maintain the retention and stability of these dentures during function. BACKGROUND: No good method with adequate clinical photographs that will empower the dentist with the necessary skill has been reported in the literature. TECHNIQUE: The technique involves making a preliminary impression in alginate to record the denture bearing area clearly, doing a border moulding procedure with excess width to fill the wide sulcus present; except in the area of anterior lingual flange from second premolar to second premolar area. The anterior lingual border of the impression is then extended sublingually without interfering with the tongue function or blocking the opening of the ducts of the sublingual gland; to maintain contact with the surrounding sublingual tissue in both protruded and retruded positions of the tongue, thereby creating good peripheral seal in the most vulnerable area in the lower denture, where the seal is easily broken during tongue movements. CONCLUSION: This impression technique, combined with arrangement of teeth on the center of the ridge in the posterior part, use of flattened cusp posterior teeth and contoured polish surface of denture base (Fisch concept) provide sufficient retention and stability to the dentures made on these severely resorbed ridges. CLINICAL SIGNIFICANCE: Patients with severely resorbed ridges cannot be rehabilitated with implants also and so normally suffer with loose lower denture fabricated with normal border moulding procedures. These patients can be rehabilitated to a great extent properly.

Orbital trauma: keep an eye for the details!

Orbital injuries are often missed out when they occur concomitantly with other facial bone fractures. Clinical examination and plain radiographic findings alone may prove inadequate in the detection of orbital floor fractures. Refined imaging techniques like CT scans and MRI are immensely helpful in the diagnosis of orbital blowout fractures. A case of 'impure' orbital blowout fracture which went undetected at the time of initial examination and its secondary surgical reconstruction is reported here. This case report emphasizes the importance of careful clinical and radiologic evaluation in orbital injuries.

Aneurysmal bone cyst of maxilla existing concurrently with a giant cell granuloma of mandible: a case report of an unusual presentation.

Aneurysmal bone cysts (ABCs) are relatively rare but well-described expansile, osteolytic psuedocysts are most commonly seen in the long bones, pelvis, and vertebra. Although a well-described lesion, much confusion exists in its pathogenesis especially of the associations of this lesion with other lesions. Approximately, one-third of patients with ABC present simultaneously with another bone pathology. The aim of this paper is to describe a rare case of simultaneously occurring ABC of the maxilla and mandible with the mandibular lesion occurring predominantly as a giant cell granuloma. The paper discusses the various histopathological variations of ABC as well as different lesions associated with ABC and tries to provide a review of different pathogenesis theories of this interesting lesion.

Cavernous lymphangioma of the lower lip--a rare case report.

Lymphangioma are rare, benign, and hamartomatous tumors of the lymphatic vessels, that show a marked predilection for the head and neck region. Very few cases of Cavernous lymphangioma have been reported to occur in the lower lip. We report a rare case of Cavernous Lymphangioma of the lower lip in a 13 years old child who was treated surgically by excision and primary closure. Postoperative wound healing was satisfactory and there was no recurrence found till the last follow up.

Le Fort I distraction osteogenesis of the edentulous maxilla.

Rehabilitation of the edentulous patient with atrophic ridges is a problem especially when compounded with a severe prognathic inter-arch relationship. It is difficult to rehabilitate these patients prosthetically without correction of the malrelation of the jaws. The established surgical techniques for correcting combined sagittal and vertical discrepancies of edentulous jaws are often prolonged and complex with attendant morbidity. This article presents a novel, simple method of correction of severe interarch sagittal discrepancy (more than 15 mm) by performing distraction osteogenesis at Le Fort I level using an internal maxillary distraction device. This method is a simple, predictable and stable option for the correction of a severe, unfavourable intermaxillary relation in edentulous patients.

Large osteochondroma of the mandibular condyle treated by condylectomy using a transzygomatic approach.

This article presents an interesting case of osteochondroma of the mandibular condyle with multiple projections in a 42-year-old man. Owing to the large size of the lesion, surgical condylectomy was performed using a transzygomatic approach.

Scaling of surface roughness in perfectly plastic disordered media.

This paper investigates surface roughness characteristics of localized plastic yield surface in a perfectly plastic disordered material. We model the plastic disordered material using perfectly plastic random spring model. Our results indicate that plasticity in a disordered material evolves in a diffusive manner until macroscopic yielding, which is in contrast to the localized failure observed in brittle fracture of disordered materials. On the other hand, the height-height fluctuations of the plastic yield surfaces generated by the spring model exhibit roughness exponents similar to those obtained in the brittle fracture of disordered materials, albeit anomalous scaling of plastic surface roughness is not observed. The local and global roughness exponents (ζ(loc) and ζ, respectively) are equal to each other, and the two-dimensional crack roughness exponent is estimated to be ζ(loc)=ζ=0.67±0.03. The probability density distribution p[Δh(ℓ)] of the height differences Δh(ℓ)=[h(x+ℓ)-h(x)] of the crack profile follows a Gaussian distribution.