A Nonlinear Three-Dimensional Finite Element Analysis of Stress Distribution and Microstrain Evaluation in Short Dental Implants with Three Different Implant-Abutment Connections in Single and Splinted Conditions in the Posterior Mandible.

Background: Stress distribution plays a vital role in the longevity and success of implant-supported prosthesis. This study evaluated the von Mises stress and microstrain in the peri-implant bone and the implant-abutment junction of short dental implants with three different implant-abutment connections in splinted and unsplinted conditions using finite element analysis (FEA). Materials and Methods: In this experimental study, nine transversely isotropic finite element models were developed, and randomly divided into three equal groups (n = 3): control, (Group AC) single-standard 4.3 × 10 mm bone level implant-supported restorations with external hexagonal (EH) connection, internal conical (IC) and internal trichannel (ITC) connection, single short implant-supported restorations (Group AT), and splinted short implant-supported restorations (Group B) for each of the three implant-abutment connections, respectively. A 200 N load was applied along the long axis of the implants and a 100 N (45°) oblique load was applied and von Mises stress and microstrain values were evaluated. Results: Single standard implants demonstrated the highest von Mises stress and microstrain values followed by single short implants and splinted short implants, respectively. Among the implant-abutment connections, the IC connection showed the highest values and the ITC connection showed the least values. Conclusion: Within the limitations of this study, it was concluded that splinting of short dental implants demonstrated lesser and more homogeneous stress and microstrain, especially on oblique loading. The microstrain values for all connections evaluated were within the physiological loading limit (200-2,500 N) and were hence considered safe for clinical use.

Impact of monsoon season rainfall spells on the ecosystem carbon exchanges of Himalayan Chir-Pine and Banj-Oak-dominated forests: a comparative assessment.

The Chir-Pine (Pinus roxburghii) and Banj-Oak (Quercus leucotrichophora)-dominated ecosystems of central Himalaya provide significant green services. However, responses of these ecosystems, with respect to ecosystem carbon flux variability, to changing microclimate are not yet studied. Since quantification of ecosystem responses to fluctuation in the microclimate, particularly rainfall, is expected to be beneficial for management of these ecosystems, this study aims (i) to quantify and compare amplitude of rainfall-induced change in the carbon fluxes of Chir-Pine and Banj-Oak-dominated ecosystems using wavelet methods, and (ii) to quantify and compare dissimilarities in the ecosystem exchanges due to varying rainfall spell and amount. Eddy covariance-based continuous daily micrometeorological and flux data, during the 2016-2017 monsoon seasons (total 244 days, 122 days of June-September), from two sites in Uttarakhand, India, are used for this purpose. We find that both Chir-Pine and Banj-Oak-dominated ecosystems are the sinks of carbon, and Chir-Pine-dominated ecosystem sequesters around 1.8 times higher carbon than the Banj-Oak. A systematic enhancement in the carbon assimilation of the Chir-Pine-dominated ecosystem is noted with increasing rainfall spell following a statistically significant power-law relationship. We have also identified a rainfall amount threshold for Chir-Pine and Banj-Oak-dominated ecosystems (10 ± 0.7 and 17 ± 1.2 mm, respectively) that resulted in highest ecosystem carbon assimilation in monsoon. The general inference of this study accentuates that Banj-Oak-dominated ecosystem is more sensitive to maximum rain within a spell whereas the Chir-Pine-dominated ecosystem is more responsive to increasing rainfall spell duration.

Singed and vinculin play redundant roles in cell migration by regulating F-actin.

INTRODUCTION: Drosophila Singed (mammalian Fascin) is an actin-binding protein that is known mainly for bundling parallel actin filaments. Among many functions of Singed, it is required for cell motility for both Drosophila and mammalian systems. Increased Fascin-1 levels positively correlate with greater metastasis and poor prognosis in human cancer. Border cell cluster, forms and migrates during Drosophila egg chamber development, shows higher expression of Singed compared with other follicle cells. Interestingly, loss of singed in border cells does not lead to any effect other than delay. RESULT: In this work, we have screened many actin-binding proteins in search of functional redundancy with Singed for border cell migration. We have found that Vinculin works with Singed to regulate border cell migration, albeit mildly. Although Vinculin is known for anchoring F-actin to the membrane, knockdown of both singed and vinculin leads to a reduced level of F-actin and changes in protrusion characteristics in border cells. We have also observed that they may act together to control microvilli length of brush border membrane vesicles and the shape of egg chambers in Drosophila. CONCLUSIONS: We may conclude that singed and vinculin work together to control F-actin and these interactions are consistent across multiple platforms.

Copper-Plumbagin Complex Produces Potent Anticancer Effects by Depolymerizing Microtubules and Inducing Reactive Oxygen Species and DNA Damage.

Here, we have synthesized a copper complex of plumbagin (Cu-PLN) and investigated its antiproliferative activities in different cancer cells. The crystal structure of Cu-PLN showed that the complex was square planar with a binding stoichiometry of 1:2 (Cu/Plumbagin). Cu-PLN inhibited the proliferation of human cervical carcinoma (HeLa), human breast cancer (MCF-7), and murine melanoma (B16F10) cells with half-maximal inhibitory concentrations (IC50) of 0.85 ± 0.05, 2.3 ± 0.1, and 1.1 ± 0.1 µM, respectively. Plumbagin inhibited the proliferation of HeLa, MCF-7, and B16F10 cells with IC50 of 7 ± 0.1, 8.2 ± 0.2, and 6.2 ± 0.4 µM, respectively, showing that Cu-PLN is a stronger antiproliferative agent than plumbagin. Interestingly, Cu-PLN showed much stronger toxicity against breast carcinoma and skin melanoma cells than noncancerous breast epithelial and skin fibroblast cells, indicating its specific cytotoxicity toward cancer cells. A short exposure of Cu-PLN triggered microtubule disassembly in cultured cancer cells, and the complex also inhibited the polymerization of purified tubulin much more strongly than plumbagin. Furthermore, Cu-PLN inhibited the binding of colchicine to tubulin. In addition to microtubule depolymerization, the antiproliferative mechanism of Cu-PLN involved induction of reactive oxygen species, reduction of the mitochondrial membrane potential, and DNA damage. Moreover, the cytotoxic effects of Cu-PLN reduced significantly in cells pre-treated with N-acetyl cysteine, suggesting that reactive oxygen species generation is crucial in Cu-PLN's mode of action. Thus, the complexation of plumbagin with copper yields a promising antitumor agent having a stronger antiproliferative activity than cisplatin, a widely used anticancer drug.

Cytotoxicity of dental cement on soft tissue associated with dental implants at different time intervals.

Background: To investigate and compare the effect of four commercially used dental cement at 24 hours, 48 hours,72 hours (h) and 6 days on the cellular response of human gingival fibroblast (HGF). Methods: 3 cement pellet samples were made for each 4-test cement (n=12). The cement used for this study were zinc phosphate (ZP), zinc oxide non-eugenol (ZOE), RelyX U200 (RU200), and glass ionomer cement (GIC). The cytotoxicity of peri-implant tissues was investigated using one commercial cell line. All processing was done following International Organization for Standardization (ISO) methods 10993-5 and 10993-12 (MTT assay Test). Cell cultures without dental cement were considered as control. Standard laboratory procedures were followed to permit cell growth and confluence over 48 hrs after sub-cultivation. Before being subjected to analysis, the cells were kept in direct contact with the cement samples for the suggested time period. To validate the results the specimens were tested three times each. Cell death and inhibition of cell growth were measured quantitatively. Results were analyzed using 1-way ANOVA (a=0.05) followed by Tukey B post hoc test. Results: The study showed the dental cement test material was cytotoxic. ZOE, ZP, GIC, and RU200 were cytotoxic in decreasing order, respectively, significantly reducing cell viability after exposure to HGF (p <0.001). Conclusions: Within the limitations of this in-vitro cellular study, results indicated that HGF were vulnerable to the test the dental cement. The highest cytotoxicity was observed in ZOE, followed by ZP, GIC, and RU200.

An In Vitro Study on the Shear Bond Strength of Feldspathic Porcelain to Nickel Chromium Alloy and Cobalt Chromium Alloy after Various Surface Treatments.

Background: To evaluate and compare the shear bond strength of feldspathic porcelain to four distinctively surface-treated Ni-Cr and Co-Cr alloys and to assess the impact of oxidation-heat treatment on porcelain to base metal alloy bond strength. Methods: 40 specimens each of nickel-chromium alloy and cobalt-chromium alloy were cast. A total of four groups of specimens were created. Group I was surface-treated by sandblasting with 50 µm alumina particles, Group II was surface-treated by sandblasting with 110 µm alumina particles, Group III and Group IV were surface-treated with 250 µm alumina particles. In Group IV, after sandblasting initially with 250 µm alumina particles, the alloys were subjected to oxidation and resandblasting with 250 µm alumina particles. Each of the specimen was coated with opaque and body porcelain and fired to a total thickness of 2 mm porcelain. A universal measuring machine was used to assess shear bond strength at a crosshead speed of 0.5 mm/min. Results: Two-way ANOVA followed by Tukey's post hoc test was used to assess the significant difference within the groups. Unpaired t-test was used for the intergroup comparison of the obtained data. The study showed that the size of the air abrasion particles used for sandblasting significantly influenced the porcelain to metal surface bond strength, with p value <0.001. The bond strength values of the two alloys tested showed no major variations. Result also showed that oxidation influences the metal-ceramic bond strength. Conclusions: The bond strength of the metal-ceramic interface is influenced by the alloy's surface treatment. The oxidation process impacts the bond strength of the metal-ceramic system.

Effect of Different Surface Treatments on the Micro-Shear Bond Strength and Surface Characteristics of Zirconia: An In Vitro Study.

Purpose: To study the effect of different surface treatments on the micro-shear bond strength and surface characteristics of zirconia. Methods: Two types of zirconia ceramics were tested: opaque (O) and translucent (T). Each type of zirconia was further allotted into four groups based on the type of surface treatment method. The four groups were: control (C), air abrasion with 110 µm Al2O3 particles (A), etching with Zircos-E Etching solution for 2 hours (E), and a combination of air abrasion and etching (AE). After the surface treatment, all specimens were ultrasonically cleaned and 10 resin cement cylinders were attached to the zirconia discs in each group. A micro-shear bond strength test was performed in a universal testing machine at a crosshead speed of 0.5 mm/min. The fracture surfaces were assessed under a compound microscope. SEM, EDAX, and AFM analyses were done for the zirconia specimens after being subjected to surface treatment. Statistical analysis for the bond strength test was done using the Shapiro-Wilk test, one-way analysis of variance (ANOVA), and Post hoc Tukey test. Results: The micro-shear bond strength values for the groups were as follows in megapascals (MPa): OC 18.96 (5.54), OA 22.66 (2.51), OE 28.48 (4.50), OAE 28.63 (4.53), TC 22.82 (5.46), TA 25.36 (5.17), TE 28.12 (4.76), and TAE 32.00 (3.47). The one-way analysis of variance (ANOVA) and post hoc Tukey HSD tests were done which showed significant results in the groups. In opaque zirconia, significant differences were seen in the etching and air abrasion with etching groups when compared with the control and air abrasion groups. There was no difference between the etching and air abrasion with etching groups. For translucent zirconia, the only significant difference was seen in the air abrasion with etching group in comparison with the control and air abrasion groups. The mode of failure was majorly adhesive. The surface topography and surface roughness showed significant differences between the groups. The EDAX results showed material loss that occurred due to sandblasting in the air abrasion groups. Conclusions: Etching with Zircos-E Etching solution significantly increased the bond strength of zirconia to resin cement when compared with other surface treatment methods. In translucent zirconia, the best results can be achieved by combining etching with air abrasion.

Biomechanical Stress Analysis of Platform Switch Implants of Varying Diameters on Different Densities of Bone.

PURPOSE: The purpose of this study was to evaluate and compare the strain developed in D2 and D3 types of bones on vertical loading by platform switch implants of different diameters. MATERIALS AND METHODS: Implants of diameters 3.25 mm, 4.2 mm, and 5.0 mm and of length 11.5 mm were taken and placed each on D2 and D3 bone models. Strain gauges were attached on the buccal and the lingual sides on each of these samples, and a vertical load of 190N was placed on the samples. The strain was recorded using a data logger. The data obtained was analysed using one-way ANOVA and post hoc Tukey test. RESULTS: In D2 and D3 bone models, 3.25 mm significantly showed greater bone strain values. The buccal side strain was higher irrespective of the implant diameter and density of bone. CONCLUSION: Within the limitations of the study, it may be concluded that the narrow diameter implant produces greater strain than 4.2 and 5.0 mm diameter implants, respectively. The buccal side consistently produced higher bone strain values.

Shikonin impedes phase separation and aggregation of tau and protects SH-SY5Y cells from the toxic effects of tau oligomers.

Tauopathies such as Alzheimer's and Parkinson's diseases involve the abnormal deposition of tau aggregates in the brain and neuronal tissues. We report that a natural naphthoquinone, shikonin, impeded the oligomerization and fibrillization of tau. The compound strongly inhibited heparin, arachidonic acid, and RNA-induced tau aggregation. Atomic force microscopy, dynamic light scattering, SDS-PAGE, and dot blot assays revealed that shikonin diminished tau oligomerization and decreased the mean size of tau oligomers. Transmission electron microscopy and atomic force microscopy analysis further showed that shikonin could suppress tau fibrillization and shorten the tau filaments. Shikonin inhibited tau droplet formation. The compound significantly reduced the aggregation rate of a tryptophan mutant (Y310W-tau) of tau. In addition, shikonin disaggregated preformed tau filaments with a half-maximal disaggregation concentration (DC50) of 6.3 ± 0.4 µM. Pre-treatment of neuroblastoma cells (SH-SY5Y) with shikonin protected the cells from the toxicity induced by tau oligomers and increased their viability. The findings imply that shikonin inhibited several steps in the tau aggregation pathways, especially the early stages, such as liquid-liquid phase separation. Therefore, shikonin is an attractive candidate for developing a therapy against tauopathy.

Cytotoxicity of Dental Cements on Soft Tissue Associated with Dental Implants.

PURPOSE: To investigate and compare the cellular host response of human gingival fibroblasts (HGF) on four currently used cements. Methods and Material. 5 cement pellet samples were made for each of the 4 test cements (n = 20). The cements used for this study were zinc phosphate, zinc oxide noneugenol (ZOE), RelyX U200, and glass ionomer cement (GIC). One commercially available cell line was used to investigate the cytotoxicity of peri-implant tissues. Direct contact cell culture testing was conducted following International Organization for Standardization (ISO) methods 10993-5 and 10993-12 (MTT assay test). Cell cultures without dental cement were considered as control. Cells were allowed to grow and confluence over 48 hours after subcultivation according to standard laboratory procedures. The cells were kept in direct contact with the cement samples for 24 hours before being subjected to analysis. All specimens were tested in triplicate to validate the results. Quantitative evaluation of cytotoxicity was done to measure cell death and inhibition of cell growth. Results were analyzed using 1-way ANOVA (a = 0.05) followed by Tukey B post hoc test. RESULTS: The results of the study showed that HGF was vulnerable to the dental cement test material. GIC, zinc phosphate, ZOE, and resin cement were cytotoxic in decreasing order, respectively, and significantly reduced the cell viability after exposure to HGF (p < 0.001). CONCLUSIONS: Within the limitations of this in vitro cellular study, results indicated that the test dental cements were cytotoxic to HGF. The highest cytotoxicity was observed in GIC followed by zinc phosphate, ZOE, and resin cement.

Contrasting Effects of Ferric and Ferrous Ions on Oligomerization and Droplet Formation of Tau: Implications in Tauopathies and Neurodegeneration.

The dysregulation of metal homeostasis is reported to enhance the aggregation of tau, a key neuronal microtubule-associated protein. Herein, we found that ferric (Fe3+) ions enhanced tau aggregation. Fe3+ and Al3+ induced tau aggregation while several trivalent metal ions such as Cr3+, La3+, and V3+ had no discernable effect on tau aggregation. Fe3+ reduced the critical concentration of tau required for the liquid-liquid phase separation (LLPS); however, Cr3+, La3+, and V3+ did not affect tau droplet formation. Dynamic light scattering, atomic force microscopic, and transmission electron microscopic analysis suggested that Fe3+ significantly increased the formation of tau oligomers and fibrils. In contrast, Fe2+ neither enhanced tau droplet formation nor increased the heparin-induced aggregation of tau. Using a tryptophan mutant (Y310W-tau) of tau, Fe3+ was found to bind to tau with four times higher affinity than Fe2+. Acrylamide quenching of the tryptophan fluorescence of Y310W-tau, 1-anilino-8-naphthalene sulfonate (ANS) fluorescence experiment, and far-UV circular dichroism analysis indicated that Fe3+ decreased the solvent exposure of the tryptophan residue, perturbed the hydrophobic surface arrangement, and disrupted the secondary structure of tau, respectively. The increase in the ß-sheet content and a subsequent decrease in the disordered content of tau due to the binding of Fe3+ may favor tau aggregation. Fe3+ may enhance and stabilize the non-covalent interactions between disordered domains of tau molecules leading to tau aggregation. The data highlighted the relationship between the dysregulation of ferric ions and neurodegenerative disorders.

Nonlinear recurrence quantification of the monsoon-season heavy rainy-days over northwest Himalaya for the baseline and future periods.

Indian summer monsoon has the characteristics of nonlinear dynamical systems. This study verifies the hypothesis that monsoon-season heavy rainy-day climatology over northwest Himalaya would exhibit certain degree of determinism, and expected to modify in its future state due to warming. Hence, recurrence quantification analysis (RQA) leading to quantification of recurrence rate (RR) and determinism (DET) are used. The monsoon-season heavy rainy-day climatologies are computed by area averaging heavy rainy-day (i.e. any day having rainfall ≥35.5 mm) of northwestern Indian Himalaya of Uttarakhand (UK), Himachal Pradesh (HP), and Union Territory of Jammu, Kashmir and Ladakh (JKL). Nonlinear characteristics are identified for a baseline period of 1970-2005 using APHRODITE data, and a bias corrected ensemble data for the future period of 2041-2099 produced using five CORDEX experiments under two warming scenarios, RCP 4.5 and 8.5. The heavy rainy-day climatology during 1970-2005 is having a correlation dimension of 1.5 indicating fractal geometry of the system in phase space. Similarly, occurrences of diagonal lines in the recurrence plots of baseline period over JKL, HP, and UK indicated the system is governed by a nonlinear chaotic attractor. A higher recurrence rate during 1970-2005 over HP (RR = 0.123, DET = 0.83) indicated greater determinism than JKL (RR = 0.119, DET = 0.78) and UK (RR = 0.121, DET = 0.75). Mean prediction time of the nonlinear dynamical trajectories controlling heavy rainy-day climatology of 1970-2005 is noted to be higher over UK. Furthermore, the RQA patterns under warmer climates of RCP 4.5 and 8.5 during 2041-2099 over UK and JKL indicate gradual reduction in the deterministic structures in the phase space. Therefore, it can be inferred that the nonlinear dynamical system governing the monsoon-season heavy rainy-day climatology is expected to lose determinism over certain regions of northwestern Himalaya under warmer climates of RCP 4.5 and 8.5.

Biocompatible Nanocomposite Tailored to Endure the Gastric Niche Renders Effective in Vitro Elimination of Intestinal Pathogenic Bacteria and Supports Adhesion by Beneficial Bacteria.

Bacteriocins produced by lactic acid bacteria (LAB) are potent therapeutic arsenals for targeting gastrointestinal pathogens and a promising alternative to antibiotics, because of their selective activity and reduced propensity to trigger collateral damage to the beneficial gut microbes. However, proteolytic inactivation in the gastric niche renders bacteriocins ineffective. The present study addresses this challenge and demonstrates that a biocompatible milk protein fraction can be leveraged to generate a robust nanocargo, which renders protection from proteolysis in the gastric milieu and facilitates delivery of the encapsulated bacteriocin pediocin. In a simulated gastric transit experiment, pediocin-loaded milk protein nanocomposite (Ped-MNC) could render a 3.0 log reduction in the viability of model gastrointestinal pathogens. Ped-MNC is nontoxic to cultured human intestinal cells (HT-29 cells) and effectively abrogates pathogenic bacteria adhering onto intestinal cells. In a combinatorial regimen, Ped-MNC and the beneficial LAB Lactobacillus plantarum DF9 could substantially reduce the levels of the pathogen Enterococcus faecalis MTCC 439 adhering onto HT-29 cells and interestingly the nanocomposite does not hinder adhesion of intestinal cells by the beneficial LAB. The developed nanocomposite holds promise as a niche specific therapeutic for selective mitigation of intestinal pathogens.

Generation of a Hydroxyapatite Nanocarrier through Biomineralization Using Cell-Free Extract of Lactic Acid Bacteria for Antibiofilm Application.

Nanoscale materials hold considerable promise in the mitigation of bacterial infections. In order to exploit nanomaterials as delivery systems in an antibacterial therapeutic paradigm, it is critical to ensure that the generated material is nontoxic. Based on the fundamental principle of biomineralization, we herein report the generation of biocompatible hydroxyapatite nanoparticles (HANPs) in the presence of proteins secreted by the lactic acid bacteria (LAB) Lactobacillus plantarum MTCC 1325, Lactobacillus plantarum CRA52, and Pediococcus pentosaceus CRA51. The biogenic HANPs were characterized by AFM, FETEM, powder XRD, DLS, and FTIR analysis. Interestingly, HANPs could also be synthesized using an ∼20 kDa protein purified from the secreted protein extract obtained from L. plantarum MTCC 1325, which suggested that this lower molecular weight protein fraction was perhaps significantly involved in biomineralization-based generation of HANPs. In order to develop a therapeutic bactericidal nanocomposite, HANPs were loaded with the antibiotic polymyxin B (PB). A Langmuir isotherm model was evident in the studies that measured adsorption of PB onto HANPs. A sustained release profile of PB from the nanocomposite was observed in buffers having varying pH and in simulated body fluid. The nanocomposite (PB-HNC) exhibited bactericidal as well as antibiofilm activity against Pseudomonas aeruginosa MTCC 2488 and was nontoxic to cultured human embryonic kidney cells.

Potential of Pyridine Amphiphiles as Staphylococcal Nuclease Inhibitor.

The present study explores the potential of pyridine-based synthetic amphiphiles C1 and C2 having 4-carbon and 12-carbon hydrophobic tails, respectively, as staphylococcal nuclease inhibitors. UV-visible titration with calf-thymus DNA (CT-DNA) revealed a hypochromic shift in the absorbance bands of C1 and C2, whereas fluorescence titration indicated a reduction in the emission intensity of the monomer bands of the amphiphiles. Interaction of deoxyribonuclease I (DNase 1) and micrococcal nuclease (MNase) with C1 or C2 led to a decrease in the emission intensity of tryptophan at λ=345 nm along with an increase in the monomer emission intensity of C1 and C2 at λ=375 nm for DNase I and excimer emission intensity at λ=470 nm for both DNase I and MNase. Scatchard's analysis indicated superior interaction of C2 with DNase I. Circular dichroism spectroscopy revealed major changes in the secondary structures of both DNase I and MNase upon interaction with the amphiphiles. A solution-based nuclease assay in conjunction with gel electrophoresis indicated amphiphile-mediated protection against nuclease-directed DNA cleavage. Interestingly, C2 could render inhibition of nuclease present in the culture supernatant of Staphylococcus aureus MTCC 96, which highlights the therapeutic prospect of the amphiphile against S. aureus.

Spermicidal efficacy of VRP, a synthetic cationic antimicrobial peptide, inducing apoptosis and membrane disruption.

Presently available contraceptives are mostly hormonal or detergent in nature with numerous side effects like irritation, lesion, inflammation in vagina, alteration of body homeostasis, etc. Antimicrobial peptides with spermicidal activity but without adverse effects may be suitable alternatives. In the present study, spermicidal activity of a cationic antimicrobial peptide VRP on human spermatozoa has been elucidated. Progressive forward motility of human spermatozoa was instantly stopped after 100 µM VRP treatment and at 350 µM, all kinds of sperm motility ceased within 20 s as assessed by the Sander-Cramer assay. The spermicidal effect was confirmed by eosin-nigrosin assay and HOS test. VRP treatment (100 µM) in human spermatozoa induced both the intrinsic and extrinsic pathways of apoptosis. TUNEL assay showed VRP treatment significantly disrupted the DNA integrity and changed the mitochondrial membrane permeability as evident from MPTP assay. AFM and SEM results depicted ultra structural changes including disruption of the acrosomal cap and plasma membrane of the head and midpiece region after treatment with 350 µM VRP. MTT assay showed after treatments with 100 and 350 µM of VRP for 24 hr, a substantial amount of Lactobacillus acidophilus (about 90% and 75%, respectively) remained viable. Hence, VRP being a small synthetic peptide with antimicrobial and spermicidal activity but tolerable to normal vaginal microflora, may be a suitable target for elucidating its contraceptive potentiality.

Micellar chemotherapeutic platform based on a bifunctional salicaldehyde amphiphile delivers a "combo-effect" for heightened killing of MRSA.

The devastating infections caused by methicillin-resistant Staphylococcus aureus (MRSA) coupled with its high resistance towards antibiotics underscores the need for an effective anti-MRSA therapeutic. The present study illustrates the use of a salicylaldehyde based bactericidal amphiphile (C1) in generating a micellar carrier that renders delivery of therapeutic antibiotics. The inherent membrane-targeting activity of C1 present in the micelle could be leveraged to counter the resistance of MRSA and enhance cellular uptake of the released antibiotics, resulting in effective elimination of the pathogen. The inherent bactericidal and antibiofilm activity of C1 was captured in FESEM analysis, solution-based assays and fluorescence microscopy. ANS-based fluorescence spectroscopy indicated that the critical micelle concentration (CMC) for C1 was 18.5 µM in water. DLS studies and FESEM analysis indicated that the average particle size for micelles based on C1 (C1M) and rifampicin-loaded C1M (C1M- R) was smaller than vancomycin-loaded C1M (C1M- V). C1M- R and C1M- V rendered sustained release of the antibiotics in physiologically relevant fluids. Notably, following interaction with MRSA for 3 h, the relative anti-MRSA activity of C1M- R and C1M- V was nearly 12-fold and 8-fold higher, respectively, as compared to the free antibiotics at equivalent concentration, highlighting the merit of leveraging the activity of C1 and the antibiotic concurrently in the micellar system. The relative cell-free antibiotic was also manifold lower in the case of C1M- R and C1M- V treated MRSA as against treatment with free antibiotics, suggesting that the amphiphilic warhead breached the membrane barrier and enhanced cellular uptake of the released antibiotics. Interestingly, C1M- R and C1M- V exhibited a high therapeutic index, being non-toxic to HEK 293 cells at concentrations higher than their minimum inhibitory concentration (MIC) against MRSA and they could be employed as an antibacterial coating to prevent MRSA biofilm formation on surgical silk sutures. The antibiotic-replete biocompatible micelles based on a self-assembling membrane-targeting amphiphile described herein represent a promising framework to integrate multiple warheads and generate a potent anti-MRSA therapeutic material.

A novel epididymal quiescence factor inhibits sperm motility by modulating NOS activity and intracellular NO-cGMP pathway.

Mature and potentially motile spermatozoa stored in cauda epididymis in an inactive state for approximately 30 days; however, during ejaculation they regain motility. To understand the actual molecular mechanism of the sperm quiescence during caudal stay, a proteinaceous quiescence factor (QF) has been purified from caprine epididymal plasma to apparent homogeneity. In the present study complete purification, detailed characterization as well as mechanistic pathway of QF has been described. QF is purified to 215-fold with 45% activity recovery. It is a 59 kDa monomeric protein with isoelectric point 5.8 and optimally active at pH 7.5. Circular dichroism spectroscopy and atomic force microscopy study confirm its α-helical secondary structure and globular tertiary conformation. QF is a thermo-stable protein as higher temperature does not alter its helical structure. N-terminal amino acid sequencing and MALDI analysis of QF did not find 100% similarity with any available protein of the database, proved its novelty. QF at 2 µM dose inhibits sperm progressive forward motility within 10 min. This motility inhibitory activity of QF is mediated by reducing NOS enzyme activity and subsequently decreasing the intracellular NO and cGMP concentration. It does not modulate intracellular Ca++ and cAMP concentration. QF has no adverse effect on DNA integrity and morphology of spermatozoa. Motility inhibitory action of QF is reversible. Thus, the role of QF in maintaining energy saving quiescence state of mature cauda spermatozoa and its reactive nitrogen species reducing activity may lead to a new direction for storage of spermatozoa and idiopathic male infertility.

Dual-label flow cytometry-based host cell adhesion assay to ascertain the prospect of probiotic Lactobacillus plantarum in niche-specific antibacterial therapy.

Host cell adhesion assays that provide quantitative insight on the potential of lactic acid bacteria (LAB) to inhibit adhesion of intestinal pathogens can be leveraged for the development of niche-specific anti-adhesion therapy. Herein, we report a dual-colour flow cytometry (FCM) analysis to assess the ability of probiotic Lactobacillus plantarum strains to impede adhesion of Enterococcus faecalis, Listeria monocytogenes and Staphylococcus aureus onto HT-29 cells. FCM in conjunction with a hierarchical cluster analysis could discern the anti-adhesion potential of L. plantarum strains, wherein the efficacy of L. plantarum DF9 was on a par with the probiotic L. rhamnosus GG. Combination of FCM with principal component analysis illustrated the relative influence of LAB strains on adhesion parameters kd and em of the pathogen and identified probiotic LAB suitable for anti-adhesion intervention. The analytical merit of the FCM analysis was captured in host cell adhesion assays that measured relative elimination of adhered LAB vis-à-vis pathogens, on exposure to either LAB bacteriocins or therapeutic antibiotics. It is envisaged that the dual-colour FCM-based adhesion assay described herein would enable a fundamental understanding of the host cell adhesion process and stimulate interest in probiotic LAB as safe anti-adhesion therapeutic agents against gastrointestinal pathogens.

A Nonbactericidal Zinc-Complexing Ligand as a Biofilm Inhibitor: Structure-Guided Contrasting Effects on Staphylococcus aureus Biofilm.

Zinc-complexing ligands are prospective anti-biofilm agents because of the pivotal role of zinc in the formation of Staphylococcus aureus biofilm. Accordingly, the potential of a thiosemicarbazone (compound C1) and a benzothiazole-based ligand (compound C4) in the prevention of S. aureus biofilm formation was assessed. Compound C1 displayed a bimodal activity, hindering biofilm formation only at low concentrations and promoting biofilm growth at higher concentrations. In the case of C4, a dose-dependent inhibition of S. aureus biofilm growth was observed. Atomic force microscopy analysis suggested that at higher concentrations C1 formed globular aggregates, which perhaps formed a substratum that favored adhesion of cells and biofilm formation. In the case of C4, zinc supplementation experiments validated zinc complexation as a plausible mechanism of inhibition of S. aureus biofilm. Interestingly, C4 was nontoxic to cultured HeLa cells and thus has promise as a therapeutic anti-biofilm agent. The essential understanding of the structure-driven implications of zinc-complexing ligands acquired in this study might assist future screening regimes for identification of potent anti-biofilm agents.