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A Pd (II)-catalyzed direct C3-(sp2)-H alkenylation of heteroarenes using benzothiazole as a directing group was successfully achieved. A wide range of 2-N-alkylpyrroles undergo an oxidative coupling with a variety of acrylates to furnish highly regio- and chemoselective E-alkenylation products at the C3 position. An important intermediate complex has been isolated and characterized so as to have an insight into the mechanism. This convenient protocol proved to be practical to access thermally activated delayed fluorescence materials (TADF). These molecules proved to be blue-emitting TADF materials (∼ms lifetime). A detailed and systematic investigation has been carried out to study the photophysical properties, and this has been further validated by the time-dependent density functional theory calculations.

The novel reactivity of a less selective and more reactive acceptor-acceptor kind of diazo pyrazolone (DIPOL) has been explored under visible light for the first time. We have successfully demonstrated the reaction of DIPOL and different allyl thioethers under blue light to construct a wide variety of products including a pesticidal analogue exclusively in excellent chemoselectivity in good to excellent yields. Moreover, possible side products emanating from ketene were not observed. This protocol works smoothly in an environmentally benign solvent under inert free conditions. The practicality of the protocol has been extended to a photoflow reaction, and also, the reaction works smoothly under the direct exposure of sunlight.

Melanins are a ubiquitous group of pigments widely acclaimed as potent free-radical scavengers. The present study proposed to harness this property of melanins for anti-inflammatory and anti-cancer applications. Pyomelanin, a potent form of melanin analogous to alkaptomelanin in humans, was derived from Pseudoalteromonasspecies and fabricated into ultra-small pyomelanin nanogranules (PNGs) by facile routes. These melanin nanogranules were characterized for various physicochemical attributes using DLS, TEM, FTIR, EPR, XRD, and TGDA. Additionally, elemental analysis and long-term particle stability study was also conducted. The ultra-small particles were ~ 5–7 nm in size through TEM with a very strict size distribution. The free-radical scavenging activity of PNG assessed through the DPPH assay was comparable with that of ascorbic acid. Significant inflammatory markers viz., cyclooxygenase, lipoxygenase, myeloperoxidase, and cellular nitrate levels estimated in lipopolysaccharide-triggered RAW264.7 cells were reduced upon PNG treatment. The cancer cell kill effect of PNG was estimated in lung carcinoma cells in comparison with normal fibroblasts, wherein percentage inhibition in cancer cells was ~ 2-fold higher than that observed in normal fibroblasts. Overall, our results demonstrate the proof-of-principle of using pyomelanin nanogranules for therapeutic applications.

ABSTRACTEnergy can be extracted from waste by chemical and biological means. There are many pathways involved in the conversion of complex carbon sources to industrially valuable products. Exploring valorization with the concept of green chemistry, focusing on transforming waste with a more environmentally compassionate product has become the new ideology. Especially efficient utilization of industrial discard as renewable feedstocks in the production of eco-friendly products are upcoming technology. The production of one such eco-friendly material, bioplastics polyhydroxyalkanoates (PHA) is discussed in this review. PHAs are naturally occurring polymers amongst various organisms and are produced intracellularly by bacterial fermentation. PHAs belong to a polyester family and, hence potentially replace hydrocarbon-based conventional polymers. Bacteria for PHA production can be grown on various renewable waste feedstocks. This process not only eliminates the waste it adds value to it. This review provides insight into the production of PHA from the feedstocks. With the possibility of synthesis of other value-added products. This can be waste-to-energy technology (WTE).

Clinical and biochemical vitamin B12(B12) deficiency is lower than anticipated in vegetarians. Extraileal absorption, such as from the colon, as well as reduced daily excretion, may be adaptive mechanisms to maintain B12homeostasis with marginal intakes.

There are limited data on in-hospital cardiac arrest (IHCA) complicating non-ST-segment-elevation myocardial infarction (NSTEMI) based on management strategy.

Quinoxalinones are a privileged class of compounds, and their structural framework is found in many bioactive compounds, natural compounds, and pharmaceuticals. Quinoxalinone is a promising scaffold for different types of functionalization, and the slight modification of the quinoxalinone skeleton is known to offer a wide range of compounds for drug discovery. Owing to the importance of the quinoxalinone scaffold, we have developed a base-mediated protocol for the C3-alkylation of quinoxalinone followed by tandem cyclization to access novel types of strenuous and fused dihalo-aziridino-quinoxalinone heterocycles via the construction of C–C and C–N bonds. The protocol proved to be simple and practical to access desired fused quinoxalinone heterocycles in excellent yields (up to 98% yield). As an application, the highly functionalized fused dihalo-aziridino-quinoxalinone molecule has been further utilized for mono-dehalogenation under visible light irradiation and selective amide reduction. Moreover, the protocol has also been demonstrated on a gram scale.

Proper functioning of any cell depends on gene regulation. Lac operon plays a major role in regulating the cell metabolism in bacterial genes. The lac genetic switch exhibits significant stochasticity. In this study, two-state model of lac-operon is considered and a mathematical model is constructed in terms of gene regulation using the Next Reaction Method. The results show that the Next Reaction Method performs much better in simulation accuracy than other methods like first reaction method, and direct method. The software operates in Python environment with other dependencies and uses the Next Reaction Method algorithm to solve stochastic equations. GPU computing was utilized to speed up the simulation performance. The number of mRNA and protein groups of consecutive packets is formed as a function of time which corresponds to when a gene is ‘on’ (no repressor is bound). The mRNA and protein data were recorded for all the simulations over a period of time and most of the simulations took six minutes to complete using a heterogeneous platform. Finally, this model includes the processes of transcription of mRNA and protein for which majority of high-throughput data is accessible.

•Residents gaining more autonomy in robotic surgery at this single institution study.•Female residents report less console time and complete fewer key steps in each case.•Differences were more prominent in higher-complexity cases.

The rapid rise of multidrug-resistant (MDR) organisms has created a critical need for alternative treatment options. Phage therapy is gaining attention as an effective way to fight bacterial infections by using lytic bacteriophages to specifically target and kill harmful bacteria. This review discusses several phage therapeutic options and emphasizes new developments in phage biology. Phage treatment has proven to be successful against MDR bacteria, as evidenced by multiple human clinical trials that indicate favorable results in treating a range of diseases caused by these pathogens. Despite these promising results, challenges such as phage resistance, regulatory hurdles, and the need for standardized treatment protocols remain. To effectively combat MDR bacterial infections, future research must focus on enhancing phage effectiveness, guaranteeing safety for human usage and incorporating phage therapy into clinical practice.

The proposed research explores Multi-Walled Carbon Nanotube’s (MWCNT’s) effect on the mechanical buckling behavior of glass fiber-enhanced thermosetting composites using UTM and the load vs displacement curve is plotted. Using the inflection point method, the critical buckling load is obtained from the load vs displacement curve for beams with three different volume fractions of MWCNT. The nonlinear finite element method is used to numerically obtain the load vs deflection curve and the numerical results are compared with the experimental results, and a close match is found with the experimental results. It is observed that the nonlinearity associated with the structure can significantly reduce the critical buckling load. The critical buckling load is found to increase and reported a 27.4% increase in buckling load with 0.3 wt.% of MWCNT which could be accounted for the increase in flexural modulus of the material.

In recent years packaging materials made of biodegradable polymers received considerable attention. The packaging system containing antimicrobial agents was one of the new methods that have an advantage over the conventional system. This work aims to fabricate and characterize antibacterial films based on chitosan (CS) and poly(vinyl alcohol) (PVA) incorporated with white turmeric powder (WT) for food packaging applications. The active films of chitosan/cetrimide/white turmeric (CCW), poly(vinyl alcohol)/cetrimide/white turmeric (PCW) and PVA/CS/CT/WT (PCCW) with varied concentrations of WT were prepared by solvent casting method. The influence of WT on the physical and structural properties of the prepared films were studied using different analytical techniques. The results of SEM analysis indicated interactions of polymer matrix with WT contributed to homogeneous, compact and smooth surface morphology. The Fourier Transform Infrared (FTIR) Spectroscopy and X-ray diffraction (XRD) analysis revealed hydrogen bond formation with a polymer matrix that improved the physical and thermal properties. The research findings revealed that there was an improvement in the tensile strength, moisture adsorption, solubility and hydrophobicity of CCW active films with an increase in WT content. At the same time, there was an improvement in inhibiting bacterial growth with increasing concentration of WT for all the fabricated films. The results showed better mechanical and antibacterial properties for PCCW films. All the films exhibited biodegradation under environmental conditions. All these results suggested WT incorporated CS and PVA based films can be used in the food packaging industry to enhance the safety and quality of the packaging material.

Background: Linagliptin (LNG) is an oral hypoglycemic agent that acts by inhibiting the enzyme dipeptidyl peptidase - 4 (DPP-4) and reduces blood sugar levels in type-II diabetic patients. To date, the literature presents few analytical methods for the determination of LNG. However, no reversed phase-high performance liquid chromatography (RP-HPLC) method has been reported for the determination of LNG in nanotransfersomes and in vitroskin permeation samples. Objective: The present study involves the development and validation of RP-HPLC method to quantify LNG in both nanotransfersomes and in vitroskin permeation and deposition samples. Methods: The chromatographic analysis was performed on Luna C18 (2) column (250 x 4.6 mm, 5μm particle size) with a mobile phase consisting of a mixture of methanol: 0.2% orthophosphoric acid (50:50, v/v) at a flow rate of 1.0 mL/min, detection wavelength of 227 nm, and column temperature of 40 °C. Results: The method was found to be specific, linear (r2≥ 0.999; 2-12 μg/mL), precise at both intra and inter-day levels (percentage relative standard deviation; % RSD < 2.00), accurate (percentage recovery 100.21-103.83%), and robust. The detection and quantification limits were 0.27 and 0.82 μg/mL, respectively. The mean % entrapment efficiency and the cumulative amount of LNG permeated across the rat skin from different transfersomal formulations ranged between 40.78 ± 2.54 % to 52.26 ± 2.15 % and 79.54 ± 16.67 to 200.74 ± 35.13 μg/cm2respectively. Conclusion: The method was successfully applied to determine the entrapment efficiency, in vitroskin permeation and deposition behavior of LNG-nanotransfersomes.

In this work, the lower extremity physiological parameters are recorded during normal walking gait, and the dynamical systems theory is applied towards its stability analysis. The human walking gait pattern of kinematic and dynamical data is approximated to periodic behavior. The embedding dimension analysis of the kinematic variable's time trace and use of Taken's theorem allows us to compute a reduced-order time series that retains the essential dynamics. In conjunction with Floquet Theory, this approach can help study the system's stability characteristics. The Lyapunov-Floquet (L-F) Transformation application results in constructing an invariant manifold resembling the form of a simple oscillator system. It is also demonstrated that the simple oscillator system, when re-mapped back to the original domain, reproduces the original system's time evolution (hip angle or knee angle, for example). A re-initialization procedure is suggested that improves the accuracy between the processed data and actual data. The theoretical framework proposed in this work is validated with the experiments using a motion capture system.

Aim: The cancer stem cells (CSCs) are known to be responsible for drug resistance and cancer relapse in the treatment of cancer. Identification and isolation of CSCs and study of their properties will play a crucial role in developing an effective drug against these targets. The aim of the study was to isolate CSCs from primary cancer by the tumorspheres enrichment method, to confirm by indirect immunofluorescence and gene expression of stem cell markers by using real-time polymerase chain reaction (RT-PCR) technique.Materials and Methods: In this in vitro study, we enriched oral CSCs through tumorsphere formation assay from seven primary cultures of OSCC patients with defined serum media. The expression and localization of the cell surface markers of CD133 and CD44 were tested by indirect immunofluorescence. Gene expression of stem cell markers such as CD44, CD133, Oct4, Sox2, and Nanog were quantified by RT-PCR technique. One-way analysis of variance was applied to analyze gene expression.Results: Tumorsphere formation has been used to isolate the CSCs from the OSCC tissue culture. Both CD133 and CD44 antibody confirmed the presence of CSCs through indirect immunofluorescence. In comparison to parental cell lines, the expression levels of CD133, CD44, Oct4, Sox2, and Nanog stem cell were significantly higher in CSC-enriched subpopulations.Conclusions: The cost-effective spheroid enrichment and the indirect immunofluorescence methods are useful for the isolation of CSCs from the primary tumor.

An efficient and straightforward method has been developed for the synthesis of β-benzyl-substituted 5-membered heterocyclic carbaldehydes via transient directing-group-enabled direct γ-C(sp3)–H arylation of 3-methylheteroarene-2-carbaldehydes. A wide range of 3-methylheteroarene carbaldehydes undergo coupling with a variety of aryl iodides, including less reactive iodo pyridine derivatives to provide a library of highly selective functionalized products in good to excellent yields. Some of these products have been successfully utilized in synthesizing useful synthetic intermediates.

A silver(I)‐catalyzed method to access spiro‐pyrrolidinedioneoxiranes has been developed. Silver hexafluoroantimonate was found to be an efficient catalyst for the epoxidation of aldehydes using donor‐/acceptor‐substituted vinyl diazosuccinimides as carbenoid precursors. The protocol has been found to be highly regio‐ and chemoselective, and works well with aromatic aldehydes containing electron‐withdrawing and ‐donating groups to afford spiro‐pyrrolidinedioneoxiranes with a broad substrate scope. Silver spiral: Silver hexafluoroantimonate was found to be an efficient catalyst for the epoxidation of aldehydes using donor‐/acceptor‐substituted vinyl diazosuccinimides as carbenoid precursors, leading to spiro‐pyrrolidinedioneoxiranes. The protocol has been found to be highly regio‐ and chemoselective, and tolerates aromatic aldehydes containing electron‐withdrawing and ‐donating groups.

Background and Aims: Porphyromonas gingivalis(P. gingivalis) is considered as an important pathogen responsible for periodontal disease which is characterized by inflammation of gingiva and destruction of periodontal ligament and alveolar bone leading to loss of tooth. Along with clinical investigations, suitable microbiological analysis needs to be performed which could provide more insight into the disease severity. We aim to quantify P. gingivalisby real-time PCR (RT-PCR) and analyze its association with demographic data including clinical parameters.Materials and Methods: The study consisted of chronic periodontitis patients (CP group) and healthy subjects (H group) with 120 samples in each group. RT-PCR was carried out by the SYBR Green assay to target 16S ribosomal ribonucleic acid species-specific region of P. gingivalis. Standard strain of P. gingivalisATCC 33277 was used as a control.Results: In the CP group, 79.16% samples were found positive for P. gingivalis, whereas 29.17% samples were positive in the H group. A significant difference was found when the prevalence was compared within males and females (P< .001 for both). In the older age groups, we found a higher rate of detection of P. gingivalis. As analyzed by Spearman’s correlation test, the number of cells of P. gingivaliswas significantly associated with probing depth (P= .02) and clinical attachment level (P = .01) in the CP group. The mean cell number of P. gingivaliswas found to be increasing with increasing levels of probing depth and clinical attachment level (P< .001 and P= .01, respectively).Conclusion: The present study reaffirms that the P. gingivalismicrobe is significantly associated with the chronic periodontitis and that its level varies with the severity of the disease. Colonization of the bacterium is significantly associated with severe forms of the disease.

Skin substitutes have shown success in complex wound reconstruction. We evaluate the use of a human acellular dermal matrix (ADM) as a viable alternative to autologous skin grafting for defects secondary to skin cancer excision. An institutional review board–approved, retrospective review of ADM-reconstructed defects secondary to skin cancer excision between 2012 and 2018 was conducted. ADM was indicated in patients with preclusive factors for general anesthesia, protracted procedure time, reluctance for additional donor site wound, and personal choice. We reviewed defect characteristics, healing time, postoperative outcomes, and patient demographics. The 228 participants (151 males, 77 females) had a median age of 72 years (range, 29–95 years), with melanoma diagnosed in 113 (49.6%), squamous cell carcinoma in 61 (26.8%), and basal cell carcinoma in 28 (12.2%) patients. The median interval to complete epidermal coverage was 42 days, with graft failure evident in six patients (2.6%). ADM is a viable, low-morbid alternative for reconstruction of defects secondary to skin cancer excision, with no donor site morbidity. With exception to complete healing time, outcomes are similar to those of autologous grafting.

This report presents with one of the rare experimental studies on the magnetic structure of hybrid framework materials, [C(ND2)3]Me2+(DCOO)3(Me = Cu, Mn, and Co), by using neutron powder diffraction. Copper guanidinium formate (CuGF) having a polar structure is a promised multiferroic member of the hybrid perovskite metal guanidinium formates and its Mn/Co analogues possessing a centrosymmetric structure. Previous investigations based on ab initiocalculations have suggested that CuGF is a multiferroic whose magnetic space groups are assignable either to Pn′a′21or Pna′21′. Our neutron experiments concurrently reject the Pna′21′ possibility and suggest the magnetic structure of CuGF as either Pna21or Pn′a′21, with both exhibiting “Type-A” magnetic ordering, free from ferromagnetic ordering along the polar-axis, with spins lying in the abplane. In contrast to CuGF, its centrosymmetric analogues Mn/CoGF are found to display “Type-G” magnetic ordering, with their spins aligned along the band caxis, respectively. The in-depth evaluation of magnetic structure of the metal guanidinium formate frameworks could be helpful in the understanding and designing of the magnetic functionalities of multiferroic hybrid perovskites and could provide an encouraging platform for the improvement of ab initiostudies.

In bone tissue engineering, the development of advanced biomaterials is crucial to address the challenges associated with the treatment and regeneration of bone defects. This study presents a novel polymeric nanocomposite consisting of Polycaprolactone (PCL) integrated with graphene oxide (GO) and cerium oxide (CeO2) nanoparticles, aiming to comprehensively explore its multifaceted potential for bone tissue engineering applications. The physicochemical properties of the composite films were analyzed. Biodegradation studies showed superior degradability to pure PCL due to added hydrophilic features and increased nanofiller-induced surface porosity. The remarkable improvement in mechanical stability is attributed to the uniform dispersion of nanofillers. Assessments of surface wettability, demonstrate a noteworthy shift towards heightened hydrophilicity, rendering a more conducive environment for biomolecules. The hybrid polymer nanocomposite exhibits remarkable hemocompatibility, antibacterial properties, antioxidant activity, and excels in facilitating apatite deposition and promoting osteogenic differentiation. Biomineralization investigations and ARS assay underscore the nanocomposite's exceptional ability to promote apatite deposition, a pivotal mineral in bone formation, owing to the altered surface properties introduced by the nanofillers. The ALP findings imply that the PCL-GO-CeO2nanocomposite actively supports and accelerates the process of osteogenic differentiation in MSCs, indicating its potential utility in bone tissue engineering. Additionally, cell culture studies conducted on HOS and rBMSC cell lines validate the nanocomposite's excellent cell viability, attachment, and proliferation, affirming its biocompatibility and potential for bone tissue engineering. Collectively, these findings establish PCL-GO-CeO2nanocomposite as a favourable option for enhancing bone regeneration.

This paper aims to apply an analytical approach to propagate parameter uncertainty through Life-Cycle Assessment (LCA) outcomes, and identify equivalence intervals that are applicable during material procurement decision-making. LCA outputs are usually presented as point estimates without accompanying confidence intervals, or associated margins of error, due to methodological ambiguity in accounting for uncertainties, be they epistemic (due to inherent randomness) or aleatory (due to lack of knowledge). This reduces the reliability of using LCA for design selection, and of decision-making during material procurement. This paper aims to fill this gap using a methodology based on Taylor’s first-order approximation to examine LCA outcomes for asphalt mixtures. The method is applied to assess the impact of variation in input flows on the estimated output emissions by propagating the parameter uncertainties through the LCA. It allows for the reporting of LCA outputs using intervals rather than point estimates. For competing designs, the overlap in intervals is used to construct equivalence intervals within which the effective impact of the designs can be considered equivalent due to expected variability in input parameters. The method is applied to construct equivalence intervals for the Global Warming Potential (GWP) of three asphalt mixtures, each having different levels of Reclaimed Asphalt Pavement (RAP). Their role in communicating LCA outcomes for decision-making is illustrated. Foreground data collected from 40 North American asphalt plants were used to estimate uncertainty in the input parameters of electricity and energy use in asphalt production. The paper adds to existing literature on uncertainties in pavement LCAs by establishing the need to incorporate uncertainty when using LCA to support decision-making during material procurement.

Complications like impaired glucose tolerance and diabetes mellitus due to iron overload need early identification in thalassemia. We studied the proportion of insulin resistance in thalassemia major patients on chronic transfusion, identified insulin resistance using homeostasis model assessment of insulin resistance (HOMA-IR) and triglyceride glucose (TYG) index, compared them and validated TYG index. In total, 73 thalassemia patients on regular transfusion for 3 years with serum ferritin >1500 ng/mL were studied. Serum ferritin, fasting blood glucose, triglycerides, and insulin levels were measured, HOMA-IR, and TYG index calculated and analyzed. Mean fasting glucose, triglyceride, and serum insulin values were 104 mg/dL, 164.18 mg/dL, and 19.6 m IU/mL, respectively. Mean serum ferritin was 5156 ng/mL. Insulin resistance was prevalent in one third of thalassemia patients and showed increase with age and serum ferritin. Insulin resistance by HOMA-IR was 32% as against 16% by TYG index with a cut-off value of 4.3. Using receiver operating charecteristic curve analysis, it was found that, by lowering the value of TYG index to 4.0215, sensitivity improved to 78.3% (from 39.13%) with specificity of 70%. Hence, we recommend a newer lower cut-off value of 4.0215 for TYG index for better sensitivity and specificity in identifying insulin resistance.

Aim:Research has demonstrated that there are multiple strains of Porphyromonas gingivaliswith varying potency to cause periodontal disease. The current study aims at using heteroduplex polymerase chain reaction (PCR) to detect the strain diversity of P. gingivalisin periodontitis lesions of varying severity in a sample of the Indian population. Materials and Methods:Subgingival plaque samples were collected from 60 individuals with varying severity of chronic periodontitis and 30 individuals with a clinically healthy periodontium. The samples were subjected to PCR analysis to identify P. gingivalis, followed by heteroduplex analysis to identify the strain diversity in a given sample. Bacterial culture was carried out as a comparative standard. Results:Of the 56 samples that were positive for P. gingivalisby PCR, 54 samples yielded eight different heteroduplex patterns. Analysis of these patterns indicated that two strains of P. gingivaliswere present in 41 individuals (45.6%) and three strains were present in 13 individuals (14.4%). Detection of P. gingivalisby PCR was significantly more in the periodontitis group as compared to the healthy group. Conclusions:Species-specific PCR and heteroduplex analysis provide a simple and accurate method to analyse the strain diversity of P. gingivalis. P. gingivaliswas detected in both healthy periodontal sites as well as sites with periodontitis. The presence of two or three P. gingivalisstrains was seen in 60% of the samples.

Marine ecosystems are highly productive as they harbor different marine microorganisms. Analysis of microbial diversity from these ecosystems will help identify new microbes having specificity for unique applications. This study deals with the culture-independent assessment of bacterial community structure and diversity of Arabian Sea sediments based on sequence analysis of 16S rDNA clone library. The sequences classified into seven phyla of bacterial domain including members of phylum Proteobacteria, Bacteriodetes, Actinobacteria, Firmicutes, Chloroflexi, Deferribacterand Acidobacterium. Most sequences were identified as phylum Proteobacteria(62.80%), with a dominance of classes Alphaproteobacteria(58.46%) and Gammaproteobacteria(38.46%). 10.47% of the sequences did not fall into any taxonomic hierarchy suggesting the possibility of unexplored/unidentified ones in marine sediments. This study adds to the limited information about the Viable But Not Culturables (VBNCs) in the sediments of Arabian Sea. Further studies are essential to understand the diversity and function in detail.

In this paper, we introduce the new concept of color Signless Laplacian energy LEc+(G). It depends on the underlying graph Gand the colors of the vertices. Moreover, we compute color signless Laplacian spectrum and the color signless Laplacian energy of families of graph with the minimum number of colors. The color signless Laplacian energy for the complement of some colored graphs is also obtained.

AbstractIn this paper, we introduce the new concept of color Signless Laplacian energy . It depends on the underlying graph and the colors of the vertices. Moreover, we compute color signless Laplacian spectrum and the color signless Laplacian energy of families of graph with the minimum number of colors. The color signless Laplacian energy for the complement of some colored graphs is also obtained.

Aims and Objectives: The oral microbial flora is highly complex and diverse with obligate anaerobic bacteria as the predominant component. Most of these are not yet cultivated/difficult to cultivate due to technical limitations. In this study, we aim to detect two novel oral bacterial species Dialister invisusand Olsenella uliby simplified and economical procedure of polymerase chain reaction (PCR) and study their association with primary and persistent endodontic infections.Material and Methods: The study involved 60 patients that included 30 patients of primary endodontic infections and 30 with persistent endodontic infections. The sample collection from the root canal was performed by universally accepted protocol by using sterile paper points. The deoxyribonucleic acid (DNA) extraction was done, followed by PCR with species specific primers. We made several changes to the protocol mentioned by original authors. We adopted a one-step protocol for amplification of bacterial DNA, omitting the 16SrDNA amplification step with universal primers.Results: It was seen that 7 (23.3 %) samples in primary endodontic infection group and 24 (80 %) samples in persistent endodontic infection group were positive for D. invisus. On the other hand, 11 (36.6 %) patients of primary endodontic infection showed positivity for O. uliin comparison to 9 (30 %) of persistent endodontic infection.Conclusion: The results from the present study showed efficient amplification of both O. uliand D. invisusin a single-step PCR. Hence, we conclude that the modified protocol used here with taq polymerase enzyme offers a faster and cheaper alternative to nested PCR without compromising the quality of amplification process.

Abstract: The increased global population has concurrently increased waste disposal, whose majority is conventional plastic. In this study, polyhydroxyalkanoates (PHA), an alternative biopolymer to conventional plastics, were extracted from bacteria Bacillus sp., using response surface methodology (RSM), a statistical approach. To design, optimize and study the relationship between the parameters (glass beads weight, incubation time, water volume, incubation temperature, and shaker speed) Box-Behnken Design of response surface methodology was applied in Design Expert 10.0 software package. The solvent method is known in PHA extraction; however, this approach is environmentally hazardous on a large scale. The current study used a physical extraction method using glass beads for bacterial cell lysis. As a characterization, FTIR, 1HNMR, and DSC confirmed the recovered polymer as Polyhydroxy butyrate (PHB). 31.53% (w/v) of PHB was recovered for 1 g/L biomass. PHB is known to be widely applied in various fields, specifically in medical applications. Genetically modified isolate, low-cost substrate, and recovery without solvent assure a cost-effective and increased PHA production. Glass beads can be reused in extraction, reducing overall production cost. Therefore, this work used a reduced amount of chemicals during extraction to recover the PHB. Thus, sustainability assures a better scope for the future promotion of PHA production in academia and industries. Highlights:

Sustainable PHA extraction alternatives, reducing environmental impact, were studied.

Glass beads are used to lyse the cell to release the PHA from the bacteria Bacillussp.

Box-Behnken design optimized extraction parameters for maximum recovery of PHA.

The cellular lysis approach yields higher recovery than solvent-method recovery and is eco-friendly. Graphical abstract: