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1.
Funct Integr Genomics ; 24(4): 120, 2024 Jul 04.
Artigo em Inglês | MEDLINE | ID: mdl-38960936

RESUMO

The Drosophila egg chamber (EC) starts as a spherical tissue at the beginning. With maturation, the outer follicle cells of EC collectively migrate in a direction perpendicular to the anterior-posterior axis, to shape EC from spherical to ellipsoidal. Filamentous actin (F-actin) plays a significant role in shaping individual migratory cells to the overall EC shape, like in every cell migration. The primary focus of this article is to unveil the function of different Actin Binding Proteins (ABPs) in regulating mature Drosophila egg shape. We have screened 66 ABPs, and the genetic screening data revealed that individual knockdown of Arp2/3 complex genes and the "capping protein ß" (cpb) gene have severely altered the egg phenotype. Arpc1 and cpb RNAi mediated knockdown resulted in the formation of spherical eggs which are devoid of dorsal appendages. Studies also showed the role of Arpc1 and cpb on the number of laid eggs and follicle cell morphology. Furthermore, the depletion of Arpc1 and cpb resulted in a change in F-actin quantity. Together, the data indicate that Arpc1 and cpb regulate Drosophila egg shape, F-actin management, egg-laying characteristics and dorsal appendages formation.


Assuntos
Actinas , Proteínas de Drosophila , Morfogênese , Animais , Proteínas de Drosophila/metabolismo , Proteínas de Drosophila/genética , Actinas/metabolismo , Actinas/genética , Feminino , Morfogênese/genética , Drosophila melanogaster/crescimento & desenvolvimento , Drosophila melanogaster/genética , Drosophila melanogaster/metabolismo , Proteínas dos Microfilamentos/metabolismo , Proteínas dos Microfilamentos/genética , Complexo 2-3 de Proteínas Relacionadas à Actina/metabolismo , Complexo 2-3 de Proteínas Relacionadas à Actina/genética , Proteínas de Capeamento de Actina/metabolismo , Proteínas de Capeamento de Actina/genética , Óvulo/metabolismo , Óvulo/crescimento & desenvolvimento
2.
Dev Dyn ; 252(7): 986-1008, 2023 07.
Artigo em Inglês | MEDLINE | ID: mdl-36912821

RESUMO

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.


Assuntos
Citoesqueleto de Actina , Actinas , Animais , Citoesqueleto de Actina/metabolismo , Actinas/metabolismo , Movimento Celular , Drosophila/metabolismo , Vinculina/metabolismo
3.
Environ Monit Assess ; 195(7): 827, 2023 Jun 09.
Artigo em Inglês | MEDLINE | ID: mdl-37294356

RESUMO

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.


Assuntos
Pinus , Quercus , Ecossistema , Quercus/fisiologia , Estações do Ano , Carbono , Árvores/fisiologia , Monitoramento Ambiental , Florestas
4.
J Cell Physiol ; 233(5): 4345-4359, 2018 05.
Artigo em Inglês | MEDLINE | ID: mdl-29150942

RESUMO

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.


Assuntos
Citoplasma/genética , Epididimo/metabolismo , Motilidade dos Espermatozoides/genética , Espermatozoides/metabolismo , Animais , Citoplasma/metabolismo , Metabolismo Energético/genética , Epididimo/crescimento & desenvolvimento , Cabras , Masculino , Cauda do Espermatozoide/metabolismo , Espermatozoides/crescimento & desenvolvimento
5.
J Cell Physiol ; 233(2): 1041-1050, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-28409819

RESUMO

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.


Assuntos
Peptídeos Catiônicos Antimicrobianos/farmacologia , Apoptose/efeitos dos fármacos , Membrana Celular/efeitos dos fármacos , Peptídeos/farmacologia , Espermicidas/farmacologia , Espermatozoides/efeitos dos fármacos , Acrossomo/efeitos dos fármacos , Acrossomo/metabolismo , Acrossomo/ultraestrutura , Membrana Celular/metabolismo , Membrana Celular/ultraestrutura , Relação Dose-Resposta a Droga , Humanos , Lactobacillus/efeitos dos fármacos , Masculino , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Viabilidade Microbiana/efeitos dos fármacos , Proteínas de Transporte da Membrana Mitocondrial/efeitos dos fármacos , Proteínas de Transporte da Membrana Mitocondrial/metabolismo , Membranas Mitocondriais/efeitos dos fármacos , Membranas Mitocondriais/metabolismo , Membranas Mitocondriais/ultraestrutura , Poro de Transição de Permeabilidade Mitocondrial , Permeabilidade , Peça Intermédia do Espermatozoide/efeitos dos fármacos , Peça Intermédia do Espermatozoide/metabolismo , Peça Intermédia do Espermatozoide/ultraestrutura , Motilidade dos Espermatozoides/efeitos dos fármacos , Espermatozoides/metabolismo , Espermatozoides/ultraestrutura , Fatores de Tempo
6.
Chembiochem ; 19(13): 1400-1408, 2018 07 04.
Artigo em Inglês | MEDLINE | ID: mdl-29645328

RESUMO

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.


Assuntos
Proteínas de Bactérias/antagonistas & inibidores , Inibidores Enzimáticos/metabolismo , Nuclease do Micrococo/antagonistas & inibidores , Piridinas/metabolismo , Tensoativos/metabolismo , Animais , Proteínas de Bactérias/química , Proteínas de Bactérias/metabolismo , Domínio Catalítico , Bovinos , DNA/química , DNA/metabolismo , Desoxirribonuclease I/antagonistas & inibidores , Desoxirribonuclease I/química , Desoxirribonuclease I/metabolismo , Inibidores Enzimáticos/química , Corantes Fluorescentes/química , Corantes Fluorescentes/metabolismo , Nuclease do Micrococo/química , Nuclease do Micrococo/metabolismo , Estrutura Molecular , Ligação Proteica , Conformação Proteica , Pirenos/química , Pirenos/metabolismo , Piridinas/química , Staphylococcus aureus/enzimologia , Tensoativos/química
7.
Microbiology (Reading) ; 163(12): 1822-1834, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-29091578

RESUMO

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.


Assuntos
Aderência Bacteriana/efeitos dos fármacos , Enterococcus faecalis/fisiologia , Citometria de Fluxo/métodos , Lactobacillus plantarum/fisiologia , Listeria monocytogenes/fisiologia , Probióticos/farmacologia , Staphylococcus aureus/fisiologia , Enterococcus faecalis/efeitos dos fármacos , Infecções por Bactérias Gram-Positivas/microbiologia , Células HT29 , Humanos , Listeria monocytogenes/efeitos dos fármacos , Listeriose/microbiologia , Infecções Estafilocócicas/microbiologia , Staphylococcus aureus/efeitos dos fármacos
8.
Chembiochem ; 18(15): 1502-1509, 2017 08 04.
Artigo em Inglês | MEDLINE | ID: mdl-28440961

RESUMO

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.


Assuntos
Benzotiazóis/farmacologia , Biofilmes/efeitos dos fármacos , Quelantes/farmacologia , Semicarbazonas/farmacologia , Staphylococcus aureus/efeitos dos fármacos , Zinco/química , Benzotiazóis/síntese química , Benzotiazóis/toxicidade , Quelantes/síntese química , Quelantes/toxicidade , Ácido Edético/farmacologia , Ácido Edético/toxicidade , Células HeLa , Humanos , Microscopia de Fluorescência , Semicarbazonas/síntese química , Semicarbazonas/toxicidade
9.
Anal Chem ; 87(13): 6974-9, 2015 Jul 07.
Artigo em Inglês | MEDLINE | ID: mdl-26059015

RESUMO

We report the synthesis of a metal-free chemosensor for highly selective sensing of pyrophosphate (PPi) anion in physiological medium. The novel phenylbenzimidazole functionalized imine containing chemosensor (L; [2,6-bis(((4-(1H-benzo[d]imidazol-2-yl)phenyl)imino) methyl)-4 methyl phenol]) could sense PPi anion through "turn-on" colorimetric and fluorimetric responses in a very competitive environment. The overall sensing mechanism is based on the aggregation-induced emission (AIE) phenomenon. Moreover, a real time in-field device application was demonstrated by sensing PPi in paper strips coated with L. Interestingly, detection of intracellular PPi ions in model human cells could also be possible by fluorescence microscopic studies without any toxicity to these cells.


Assuntos
Difosfatos/análise , Metais/química , Ânions , Células HeLa , Humanos , Microscopia de Força Atômica
10.
Int J Biometeorol ; 59(3): 325-38, 2015 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24899395

RESUMO

Using micrometeorological techniques to measure greenhouse gas emissions from differently treated adjacent plots is a promising avenue to verify the effect of mitigation strategies at the field scale. In pursuing such an approach, it is crucial to accurately characterize the source area of the fluxes measured at each sampling point. Hence, a comprehensive footprint analysis method is required so that emission rates can be obtained for a specific field within a biochemically heterogeneous area. In this study, a footprint analysis method is developed to estimate the emission for an experiment where the flux of N2O is measured from several control and treated plots. The emission rate of an individual plot is estimated using an inverse footprint fraction approach where the footprint fractions are obtained from an analytical footprint model. A numerical solution for obtaining the background flux for such a multiplot measurement system is also provided. Results of the footprint analysis method are assessed, first, by comparing footprint fractions obtained from both an analytical footprint model and a "forward" simulation of a backward Lagrangian stochastic (bLs) model; and second, by comparing the emission rates of a control plot obtained from the footprint analysis method and from the "backward" simulation of the bLs model. It is found that the analytical footprint fractions compare well with the values obtained from the bLs model (correlation coefficient of 0.58 and 0.66 within p value <0.001). An average of 4.3 % of the measured fluxes is found to be contributed by sources outside the measured area and, excluding this outside area contribution to the measured flux, footprint corrected emission rates within the defined domain are found to increase by 2.1 to 5.8 % of the measured flux. Also, the proposed method of emission rate estimation is found to work well under a wide range of atmospheric stability.


Assuntos
Poluentes Atmosféricos/análise , Monitoramento Ambiental/métodos , Modelos Teóricos , Óxido Nitroso/análise , Indústria de Laticínios , Nova Zelândia , Processos Estocásticos
11.
Anal Chem ; 85(17): 8369-75, 2013 Sep 03.
Artigo em Inglês | MEDLINE | ID: mdl-23905654

RESUMO

A diformyl-quinoline based receptor (L1) exhibits selective colorimetric and fluorometric sensing of Zn(2+) in aqueous medium at pH 7.4 based on the intraligand charge transfer (ICT) process. The in situ formed phenoxo-bridged complex, L1·2Zn can selectively and specifically sense PPi among all the other biologically important anions including ATP through reversible binding. The detection limit for Zn(2+) and PPi were found to be approximately 56 and 2 ppb, respectively. The unique selectivity of the PPi by the L1-Zn ensemble could be used as an analytical tool to probe PPi generation in a prototype polymerase chain reaction (PCR) setup and track DNA amplification with higher sensitivity as compared to conventional agarose gel electrophoresis. Interestingly, the principle of PPi estimation in PCR rendered rapid estimation of bacterial cell numbers with a limit of detection of 10 CFU of Escherichia coli MTCC 433 in as early as 10 PCR cycles. The proposed method of PPi sensing offers interesting application potential in PCR-based rapid diagnostics for pathogenic agents and microbiological quality control.


Assuntos
DNA Bacteriano/análise , Difosfatos/química , Escherichia coli/química , Escherichia coli/citologia , Reação em Cadeia da Polimerase/métodos , Zinco/química , Contagem de Células/métodos
12.
ACS Omega ; 8(3): 3221-3235, 2023 Jan 24.
Artigo em Inglês | MEDLINE | ID: mdl-36713695

RESUMO

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.

13.
Int J Dent ; 2023: 8851098, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37885810

RESUMO

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.

14.
F1000Res ; 12: 1342, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-38826571

RESUMO

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.


Assuntos
Cimentos Dentários , Implantes Dentários , Fibroblastos , Gengiva , Humanos , Cimentos Dentários/toxicidade , Fibroblastos/efeitos dos fármacos , Gengiva/efeitos dos fármacos , Gengiva/citologia , Implantes Dentários/efeitos adversos , Fatores de Tempo , Proliferação de Células/efeitos dos fármacos , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Teste de Materiais
15.
Int J Biol Macromol ; 204: 19-33, 2022 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-35120943

RESUMO

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.


Assuntos
Doença de Alzheimer , Naftoquinonas , Tauopatias , Doença de Alzheimer/metabolismo , Humanos , Naftoquinonas/farmacologia , Neurônios/metabolismo , Proteínas tau/metabolismo
16.
Int J Dent ; 2022: 5972259, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35251181

RESUMO

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.

17.
Int J Dent ; 2022: 1546802, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35464102

RESUMO

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.

18.
Int J Dent ; 2022: 2557127, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35677808

RESUMO

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.

19.
Int J Dent ; 2022: 4916464, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35096067

RESUMO

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.

20.
Sci Total Environ ; 789: 147754, 2021 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-34051505

RESUMO

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.


Assuntos
Clima , Chuva , Humanos , Dinâmica não Linear , Recidiva , Estações do Ano
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