High-throughput screening to evaluate optimum coagulation conditions via colloidal stability analysis.

Current methods of optimizing the coagulant dosage in wastewater treatment processes typically rely on the use of labor- and material-intensive jar testers, which are inadequate when coagulation processes require frequent adjustments due to variations in properties of the incoming feed. Analytical centrifuges (ACs) employ an integrated optics system that simultaneously monitors the position of the boundary between two separating phases in multiple samples of fairly low volumes (∼2 mL) - thus it was expected that ACs would be ideally suited to study the stability and settling kinetics of coagulation treatment processes. In this study, wastewater samples from a biogas generation facility (known as centrate) were collected in February 2022 (Batch A) and July 2022 (Batch B). A comprehensive screening of the treatment performance for Batch B was conducted at three pHs (5, 6, and 7) and nine concentrations of ferric chloride (0-500 mg-Fe3+/L) - it was found that the front-tracking profiles measured by the integrated optics system could be used to identify the minimal coagulation conditions needed to transition from slow to rapid settling. While the settling velocity was found to be well correlated with the instability index, a dimensionless number between 0 and 1 (where values closer to 1 indicate better separation), it was determined that the percentage of COD removal from the centrate samples increased up to an instability index of approximately 0.5 and then plateaued. Finally, it was found that the front-tracking profiles could be used to estimate the volume of sludge produced at various coagulation conditions. Thus, the results from this study establish ACs as an important screening tool for rapid evaluation of treatment performance while consuming minimal material and time - in this study, a total of 132 screening experiments were conducted using approximately ∼11 L of centrate and ∼6 hours of operator time.

Strain and crystallite size controlled ordering of Heusler nanoparticles having high heating rate for magneto-thermal application.

Heusler compound nanoparticles with good structural ordering need to be investigated as a potential material class for magneto-thermal applications requiring heat generation in presence of an oscillating magnetic field. Here, we report an important finding of a structural parameter related to the product of the strain and the coherent crystallite size, that can be used to efficiently control the structural ordering and the magnetic property of the Heulser compound nanoparticles. The optimization of this product parameter is found to enhance both the structural ordering and magnetic transition temperature in Co2FeSn Heusler nanoparticles. Furthermore, using magnetic hyperthermia measurements we demonstrate the possibility of heat generation using Heusler compound nanoparticles comparable to that of conventional magnetic nanoparticles. This shall lead to the development of Heulser compounds for similar applications.

Comprehensive Lipidomic and Metabolomic Analysis for Studying Metabolic Changes in Lung Tissue Induced by a Vaccine against Respiratory Syncytial Virus.

Respiratory syncytial virus (RSV) is the leading cause of acute lower respiratory infections in young children. Although the disease may be severe in immunocompromised, young, and elderly people, there is currently no approved vaccine. We previously reported the development and immunological assessment of a novel intranasal vaccine formulation consisting of a truncated version of the RSV fusion protein (ΔF) combined with a three-component adjuvant (TriAdj). Now, we aim to investigate the mechanism of action of the ΔF/TriAdj formulation by searching for metabolic alterations caused by intranasal immunization and the RSV challenge. We carried out untargeted lipidomics and submetabolome profiling (carboxylic acids and amine/phenol-containing metabolites) of lung tissue from ΔF/TriAdj-immunized and nonimmunized, RSV-challenged mice. We observed significant changes of lipids involved in the lung surfactant layer for the nonimmunized animals compared to healthy controls but not for the immunized mice. Metabolic pathways involving the synthesis and regulation of amino acids and unsaturated fatty acids were also modulated by immunization and the RSV challenge. This study illustrates that lipidomic and metabolomic profiling could provide a more comprehensive understanding of the immunological and metabolic alterations caused by RSV and the modulation effected by the ΔF/TriAdj formulation.

Selection of adjuvants for vaccines targeting specific pathogens.

INTRODUCTION: Adjuvants form an integral component in most of the inactivated and subunit vaccine formulations. Careful and proper selection of adjuvants helps in promoting appropriate immune responses against target pathogens at both innate and adaptive levels such that protective immunity can be elicited. Areas covered: Herein, we describe the recent progress in our understanding of the mode of action of adjuvants that are licensed for use in human vaccines or in clinical or pre-clinical stages at both innate and adaptive levels. Different pathogens have distinct characteristics, which require the host to mount an appropriate immune response against them. Adjuvants can be selected to elicit a tailor-made immune response to specific pathogens based on their unique properties. Identification of biomarkers of adjuvanticity for several candidate vaccines using omics-based technologies can unravel the mechanism of action of modern and experimental adjuvants. Expert opinion: Adjuvant technology has been revolutionized over the last two decades. In-depth understanding of the role of adjuvants in activating the innate immune system, combined with systems vaccinology approaches, have led to the development of next-generation, novel adjuvants that can be used in vaccines against challenging pathogens and in specific target populations.

Metabolomic and Immunological Profiling of Respiratory Syncytial Virus Infection after Intranasal Immunization with a Subunit Vaccine Candidate.

Respiratory syncytial virus (RSV) is a significant cause of mortality and morbidity in infants, the elderly, immunocompromised individuals, and patients with congenital heart diseases. Despite extensive efforts, a vaccine against RSV is still not available. We have previously reported the development of a subunit vaccine (ΔF/TriAdj) composed of a truncated version of the fusion protein (ΔF) and a polymer-based combination adjuvant (TriAdj). We compared inflammatory responses of ΔF/TriAdj-vaccinated and unvaccinated mice following intranasal challenge with RSV. Rapid and early inflammatory responses were observed in lung samples from both groups but modulated in the vaccinated group 7 days after the viral challenge. The underlying mechanism of action of ΔF/TriAdj was further studied through LC-MS-based metabolomic profiling by using 12C- or 13C-dansyl labeling for the amine/phenol submetabolome. RSV infection predominantly affected the amino acid biosynthesis pathways and urea cycle, whereas ΔF/TriAdj modulated the concentrations of almost all of the altered metabolites. Tryptophan metabolites were significantly affected, including indole, l-kynurenine, xanthurenic acid, serotonin, 5-hydroxyindoleacetic acid, and 6-hydroxymelatonin. The results from the present study provide further mechanistic insights into the mode of action of this RSV vaccine candidate and have important implications in the design of metabolic therapeutic interventions.

Pathway driven self-assembly and living supramolecular polymerization in an amyloid-inspired peptide amphiphile.

Peptide 1 with an Aß42 amyloid nucleating core demonstrates step-wise self-assembly in water. Variation of temperature or solvent composition arrests the self-assembly to give metastable nanoparticles, which undergo self-assembly on gradual increase in temperature and eventually produce kinetically controlled nanofibers and thermodynamically stable twisted helical bundles. Mechanical agitation of the fibers provided access to short seeds with narrow polydispersity index, which by mediation of seeded supramolecular polymerization establishes perfect control over the length of the nanofibers. Such pathway dependence and the length control of the supramolecular peptide nanofibers is exploited to tune the mechanical strength of the resulting hydrogel materials.

The respiratory syncytial virus fusion protein formulated with a polymer-based adjuvant induces multiple signaling pathways in macrophages.

Respiratory syncytial virus (RSV) causes acute respiratory tract infections in infants, the elderly and immunocompromised individuals. No licensed vaccine is available against RSV. We previously reported that intranasal immunization of rodents and lambs with a RSV vaccine candidate (ΔF/TriAdj) induces protective immunity with a good safety profile. ΔF/TriAdj promoted innate immune responses in respiratory mucosal tissues in vivo, by local chemokine and cytokine production, as well as infiltration and activation of immune cells including macrophages. The macrophage is an important cell type in context of both innate and adaptive immune responses against RSV. Therefore, we characterized the effects of ΔF/TriAdj on a murine macrophage cell line, RAW264.7, and bone marrow-derived macrophages (BMMs). A gene expression study of pattern recognition receptors (PRRs) revealed induction of endosomal and cytosolic receptors in RAW264.7 cells and BMMs by ΔF/TriAdj, but no up-regulation by ΔF in PBS. As a secondary response to the PRR gene expression, induction of several chemokines and pro-inflammatory cytokines, as well as up-regulation of MHC-II and co-stimulatory immune markers, was observed. To further investigate the mechanisms involved in ΔF/TriAdj-mediated secondary responses, we used relevant signal transduction pathway inhibitors. Based on inhibition studies at both transcript and protein levels, JNK, ERK1/2, CaMKII, PI3K and JAK pathways were clearly responsible for ΔF/TriAdj-mediated chemokine and pro-inflammatory cytokine responses, while the p38 and NF-κB pathways appeared to be not or minimally involved. ΔF/TriAdj induced IFN-ß, which may participate in the JAK-STAT pathway to further amplify CXCL-10 production, which was strongly up-regulated. Blocking this pathway by a JAK inhibitor almost completely abrogated CXCL-10 production and caused a significant reduction in the cell surface expression of MHC-II and co-stimulatory immune markers. These data demonstrate that ΔF/TriAdj induces multiple signaling pathways in macrophages.

Formulation of the respiratory syncytial virus fusion protein with a polymer-based combination adjuvant promotes transient and local innate immune responses and leads to improved adaptive immunity.

Respiratory syncytial virus (RSV) causes serious upper and lower respiratory tract infections in newborns and infants. Presently, there is no licensed vaccine against RSV. We previously reported the safety and efficacy of a novel vaccine candidate (ΔF/TriAdj) in rodent and lamb models following intranasal immunization. However, the effects of the vaccine on the innate immune system in the upper and lower respiratory tracts, when delivered intranasally, have not been characterized. In the present study, we found that ΔF/TriAdj triggered transient production of chemokines, cytokines and interferons in the nasal tissues and lungs of BALB/c mice. The types of chemokines produced were consistent with the populations of immune cells recruited, i.e. dendritic cells, macrophages and neutrophils, in the nose-associated lymphoid tissue (NALT), lung and their draining lymph nodes of the ΔF/TriAdj-immunized group. In addition, ΔF/TriAdj stimulated cellular activation with generation of mucosal and systemic antibody responses, and conferred complete protection from viral infection in the lungs upon RSV challenge. The effect of ΔF/TriAdj was short-lived in the nasal tissues and more prolonged in the lungs. In addition, both innate and adaptive immune responses were lower when mice were immunized with ΔF alone. These results suggest that ΔF/TriAdj modulates the innate mucosal environment in both upper and lower respiratory tracts, which contributes to robust adaptive immune responses and long-term protective efficacy of this novel vaccine formulation.

IL-12p40 gene-deficient BALB/c mice exhibit lower weight loss, reduced lung pathology and decreased sensitization to allergen in response to infection with pneumonia virus of mice.

Respiratory syncytial virus (RSV) is a major cause of bronchiolitis and pneumonia in infants and pneumonia virus of mice (PVM) causes similar disease. BALB/c mice are highly susceptible, while C57BL/6 mice are more resistant to PVM. IL-12 was significantly more up-regulated in response to PVM infection in BALB/c than in C57BL/6 mice. IL-12p40-deficient neonatal and adult BALB/c mice showed significantly less weight loss than wild-type mice after PVM challenge. The percentage of regulatory T cells, as well as IFN-ß and IL-18 expression, was higher in the lungs of both neonatal and adult IL-12p40-/- mice. Adult IL-12p40-/- mice also showed enhanced TGF-ß and IL-10 expression and reduced inflammatory responses. Furthermore, IL-12p40-/- mice showed decreased sensitization to inhaled cockroach antigen after PVM infection when compared to wild-type mice. In conclusion, these data suggest that a depressed regulatory capacity in BALB/c mice to PVM infection results in enhanced immunopathology and sensitization to allergen.

Efficacy of Photodynamic Therapy and Lasers as an Adjunct to Scaling and Root Planing in the Treatment of Aggressive Periodontitis - A Clinical and Microbiologic Short Term Study.

INTRODUCTION: Aggressive periodontitis comprises a group of rare, severe, rapidly progressive form of periodontitis. Conventional treatment includes mechanical debridement augmented with adjunctive antimicrobial therapy. Development of antibiotic resistance has led to use of lasers. Photodynamic therapy (PDT) is a novel non-invasive therapeutic approach with increased site and pathogen specificity. This study compares PDT and Lasers as an adjunct to conventional Scaling in the treatment of patients with aggressive periodontitis. MATERIALS AND METHODS: Fifteen untreated aggressive periodo-ntitis patients were randomly assigned in a split mouth design for one of the following treatment modalities: 1) SRP alone; (2) SRP + Diode Laser irradiation with 810 nm at 1W, continuous mode for 30 sec per tooth; (3) SRP + PDT on "0" day; (4) SRP + PDT on "0", 7(th) and 21(st) day. The clinical parameters included PI, BOP, PPD, CAL recorded at the baseline & 3(rd) month. The site with greatest probing pocket depth (PPD) was selected from each quadrant for bacterial sampling and cultured for Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis & Prevotella intermedia. RESULTS: Statistically significant reduction in clinical & microbial parameters was seen. Sites 4 showed a greater reduction compared to other groups. CONCLUSION: Photodynamic therapy is a valuable treatment modality adjunctive to conventional scaling and root planing.

Successful Therapy of Murine Visceral Leishmaniasis with Astrakurkurone, a Triterpene Isolated from the Mushroom Astraeus hygrometricus, Involves the Induction of Protective Cell-Mediated Immunity and TLR9.

In our previous report, we showed that astrakurkurone, a triterpene isolated from the Indian mushroom Astraeus hygrometricus (Pers.) Morgan, induced reactive oxygen species, leading to apoptosis in Leishmania donovani promastigotes, and also was effective in inhibiting intracellular amastigotes at the 50% inhibitory concentration of 2.5 µg/ml. The aim of the present study is to characterize the associated immunomodulatory potentials and cellular activation provided by astrakurkurone, leading to effective antileishmanial activity in vitro and in vivo Astrakurkurone-mediated antileishmanial activity was evaluated by real-time PCR and flow cytometry. The involvement of Toll-like receptor 9 (TLR9) was studied by in vitro assay in the presence of a TLR9 agonist and antagonist and by in silico modeling of a three-dimensional structure of the ectodomain of TLR9 and its interaction with astrakurkurone. Astrakurkurone caused a significant increase in TLR9 expression of L. donovani-infected macrophages along with the activation of proinflammatory responses. The involvement of TLR9 in astrakurkurone-mediated amastigote killing has been evidenced from the fact that a TLR9 agonist (CpG, ODN 1826) in combination with astrakurkurone enhanced the amastigote killing, while a TLR9 antagonist (bafilomycin A1) alone or in combination with astrakurkurone curbed the amastigote killing, which could be further justified by in silico evidence of docking between mouse TLR9 and astrakurkurone. Astrakurkurone was found to reduce the parasite burden in vivo by inducing protective cytokines, gamma interferon and interleukin 17. Moreover, astrakurkurone was nontoxic toward peripheral blood mononuclear cells of immunocompromised patients with visceral leishmaniasis. Astrakurkurone, a nontoxic antileishmanial, enhances the immune efficiency of host cells, leading to parasite clearance in vitro and in vivo.

Comparative evaluation of the efficacy of curcumin gel with and without photo activation as an adjunct to scaling and root planing in the treatment of chronic periodontitis: A split mouth clinical and microbiological study.

AIMS AND OBJECTIVES: Harnessing Mother Nature's bountiful remedies for rejuvenation has been in vogue since time immemorial. Turmeric contains the polyphenol Curcumin in its rhizome. It produces reactive oxygen species (ROS) with visible light irradiation as photodynamic therapy (PDT) - which validates its use in the treatment of periodontitis. This study compares Curcumin and Curcumin PDT as an adjunct to conventional Scaling and Root Planing (SRP) with SRP alone in the treatment of patients with chronic periodontitis. MATERIALS AND METHODS: Sixty sites in fifteen untreated chronic periodontitis patients were randomly assigned in a split mouth design for one of the treatment modalities; 1) Scaling and root planing (SRP) alone, (2) SRP + Curcumin application for 5 min, (3) SRP + Curcumin application for 5 min + irradiation with blue light emitting diode of wavelength 470 nm for 5 min. (Curcumin PDT) on 0 day.(4) SRP + Curcumin PDT on "0", 7(th) and 21(st) day. The clinical parameters included plaque index (PI), bleeding on probing (BOP) measured by sulcus bleeding index (SBI), probing pocket depth (PPD), clinical attachment level (CAL) recorded at the baseline & 3(rd) month. The site with greatest probing pocket depth (PPD) was selected from each quadrant for bacterial sampling and culturing for Aggregatibacter actinomycetemcomitans (Aa) and other black pigment producing microorganisms (BPB) like Porphyromonas gingivalis & Prevotella intermedia. CONCLUSION: The present study showed that Curcumin photodynamic therapy is a valuable treatment modality adjunctive to conventional scaling and root planing over Curcumin application. Moreover, multiple adjunctive applications of photodynamic therapy are more beneficial than single application in reducing clinical & microbiological parameters.

Blunted inflammatory and mucosal IgA responses to pneumonia virus of mice in C57BL/6 neonates are correlated to reduced protective immunity upon re-infection as elderly mice.

Respiratory syncytial virus is a major cause of bronchiolitis in infants and pneumonia virus of mice (PVM) causes similar disease in mice. The impact of PVM infection in BALB/c and C57BL/6 neonates, and upon re-infection as elderly mice, was compared. As previously shown for adult mice, PVM caused more disease in BALB/c than in C57BL/6 neonates. After PVM-15 infection BALB/c neonates showed higher production of inflammatory mediators, more influx of plasmacytoid dendritic cells and higher IFN-α expression, and more IgA in the lungs than C57BL/6 neonates. After re-infection as elderly, BALB/c mice developed virus neutralizing antibodies in serum and lung, and were protected from clinical disease, whereas C57BL/6 mice did not develop an anamnestic response and were not protected. These results suggest that an effective local innate response, as well as priming of mucosal adaptive responses in neonates after PVM-15 infection is correlated to decreased susceptibility and protection upon re-infection.

Vascular endothelial growth factor levels in gingival crevicular fluid before and after periodontal therapy.

INTRODUCTION: Of the various cytokines and growth factors regulating angiogenesis, the most potent agent acting on vascular endothelium is vascular endothelial growth factor(VEGF). The present study aims to access VEGF concentration in periodontal diseases and the effect of periodontal therapy on its concentration in gingival crevicular fluid (GCF). MATERIALS AND METHODS: Ninety five subjects (47 females and 48 males) 25- 47 y of age were divided into healthy (group-1), gingivitis (group-2), and periodontitis (group-3). GCF samples were collected using micro-capillary pipettes & were transferred immediately to plastic vials and stored at -70˚C until the time of the assay. The concentration of VEGF was determined using commercially available ELISA kit. RESULTS: The mean VEGF concentration was highest in periodontitis patients (Group 3) (88.08±8.04pg/ml), with lowest in healthy patients (Group 1). VEGF levels reduced significantly after therapy in Groups 2 and 3. CONCLUSION: VEGF levels in GCF had a significant correlation with both periodontal disease progression and healing after therapy.

A wavelet-based multiresolution approach to solve the stereo correspondence problem using mutual information.

In this correspondence, we propose a wavelet-based hierarchical approach using mutual information (MI) to solve the correspondence problem in stereo vision. The correspondence problem involves identifying corresponding pixels between images of a given stereo pair. This results in a disparity map, which is required to extract depth information of the relevant scene. Until recently, mostly correlation-based methods have been used to solve the correspondence problem. However, the performance of correlation-based methods degrades significantly when there is a change in illumination between the two images of the stereo pair. Recent studies indicate MI to be a more robust stereo matching metric for images affected by such radiometric distortions. In this short correspondence paper, we compare the performances of MI and correlation-based metrics for different types of illumination changes between stereo images. MI, as a statistical metric, is computationally more expensive. We propose a wavelet-based hierarchical technique to counter the increase in computational cost and show its effectiveness in stereo matching.

Study of oxidative stress and trace element levels in patients with alcoholic and non-alcoholic coronary artery disease.

Alcohol consumption induces oxidative stress, and leads to lipid peroxidation. These effects have been linked to alcohol-related toxicity and diseases are considered relevant to alcohol-atherosclerosis interrelationship. Deficiency of many antioxidants and trace elements may impair the antioxidant defense leading to ethanol induced oxidative stress. In the present study, our aim was to investigate the lipid peroxidation, lipid profile, antioxidant enzymes and trace elements in patients with and without alcoholic coronary artery disease (CAD). Our study included 61 patients suffering from CAD, 124 patients suffering from alcoholic CAD with high to moderate alcohol intake, 75 controls were randomly selected for our study. Increased serum lipid peroxidation, total cholesterol, triglycerides, LDL cholesterol and copper levels were high while levels of HDL cholesterol, glutathione peroxdiase, superoxide dismutase, trace elements like Selenium and Zinc were low in high alcoholic CAD patients compared with moderate and non alcoholic CAD patients. The results obtained from present study indicate that high alcohol intake predicts low antioxidant enzyme and that trace element may contribute to the increased susceptibility for the development of CAD.

Extracellular biosynthesis of magnetite using fungi.

The development of synthetic processes for oxide nanomaterials is an issue of considerable topical interest. While a number of chemical methods are available and are extensively used, the collaborations are often energy intensive and employ toxic chemicals. On the other hand, the synthesis of inorganic materials by biological systems is characterized by processes that occur at close to ambient temperatures and pressures, and at neutral pH (examples include magnetotactic bacteria, diatoms, and S-layer bacteria). Here we show that nanoparticulate magnetite may be produced at room temperature extracellularly by challenging the fungi, Fusarium oxysporum and Verticillium sp., with mixtures of ferric and ferrous salts. Extracellular hydrolysis of the anionic iron complexes by cationic proteins secreted by the fungi results in the room-temperature synthesis of crystalline magnetite particles that exhibit a signature of a ferrimagnetic transition with a negligible amount of spontaneous magnetization at low temperature.