A study was undertaken to evaluate the effect of artificial aging through steam and thermal treatment as influencing the shear bond strength between three different commercially available zirconia core materials, namely, Upcera, Ziecon, and Cercon, layered with VITA VM9 veneering ceramic using Universal Testing Machine. The mode of failure between zirconia and ceramic was further analyzed as adhesive, cohesive, or mixed using stereomicroscope. X-ray diffraction and SEM (scanning electron microscope) analysis were done to estimate the phase transformation (m-phase fraction) and surface grain size of zirconia particles, respectively. The purpose of this study was to simulate the clinical environment by artificial aging through steam and thermal treatment so as the clinical function and nature of the bond between zirconia and veneering material as in a clinical trial of 15 years could be evaluated.
A biomimetic minimalist model membrane was used to study the mechanism and kinetics of cell-free in vitro HIV-1 Gag budding from a giant unilamellar vesicle (GUV). Real-time interaction of Gag, RNA, and lipid, leading to the formation of mini-vesicles, was measured using confocal microscopy. Gag forms resolution-limited punctae on the GUV lipid membrane. Introduction of the Gag and urea to a GUV solution containing RNA led to the budding of mini-vesicles on the inside surface of the GUV. The GUV diameter showed a linear decrease in time due to bud formation. Both bud formation and decrease in GUV size were proportional to Gag concentration. In the absence of RNA, addition of urea to GUVs incubated with Gag also resulted in subvesicle formation. These observations suggest the possibility that clustering of GAG proteins leads to membrane invagination even in the absence of host cell proteins. The method presented here is promising, and allows for systematic study of the dynamics of assembly of immature HIV and help classify the hierarchy of factors that impact the Gag protein initiated assembly of retroviruses such as HIV.
Electronic supplementary material
The online version of this article (doi:10.1007/s10867-014-9370-z) contains supplementary material, which is available to authorized users.
HIV-1 Gag; Lipid Gag interaction; Gag multimerization; Vesicle budding; Vesicle formation rate
Nitroglycerin (NTG) markedly enhances nitric oxide (NO) bioavailability. However, its ability to mimic the anti-inflammatory properties of NO remains unknown. Here, we examined whether NTG can suppress endothelial cell (EC) activation during inflammation and developed NTG nanoformulation to simultaneously amplify its anti-inflammatory effects and ameliorate adverse effects associated with high-dose NTG administration. Our findings reveal that NTG significantly inhibits human U937 cell adhesion to NO-deficient human microvascular ECs in vitro through an increase in endothelial NO and decrease in endothelial ICAM-1 clustering, as determined by NO analyzer, microfluorimetry, and immunofluorescence staining. Nanoliposomal NTG (NTG-NL) was formulated by encapsulating NTG within unilamellar lipid vesicles (DPhPC, POPC, Cholesterol, DHPE-Texas Red at molar ratio of 6:2:2:0.2) that were ~155 nm in diameter and readily uptaken by ECs, as determined by dynamic light scattering and quantitative fluorescence microscopy, respectively. More importantly, NTG-NL produced a 70-fold increase in NTG therapeutic efficacy when compared with free NTG while preventing excessive mitochondrial superoxide production associated with high NTG doses. Thus, these findings, which are the first to reveal the superior therapeutic effects of an NTG nanoformulation, provide the rationale for their detailed investigation for potentially superior vascular normalization therapies.
Back pain is a common clinical problem and is the frequent complaint for referral of lumbar spine magnetic resonance imaging (MRI). Coronal short tau inversion recovery sequence (STIR) can provide diagnostically significant information in small percentage of patients.
Materials and Methods:
MRI examinations of a total of 350 patients were retrospectively included in the study. MR sequences were evaluated in two settings. One radiologist evaluated sagittal and axial images only, while another radiologist evaluated all sequences, including coronal STIR sequence. After recording the diagnoses, we compared the MRI findings in two subsets of patients to evaluate additional merit of coronal STIR imaging.
With addition of coronal STIR imaging, significant findings were observed in 24 subjects (6.8%). Twenty-one of these subjects were considered to be normal on other sequences and in three subjects diagnosis was changed with the addition of coronal STIR. Additional diagnoses on STIR included sacroiliitis, sacroiliac joint degenerative disease, sacral stress/insufficiency fracture/Looser's zones, muscular sprain and atypical appendicitis.
Coronal STIR imaging can provide additional diagnoses in a small percentage of patients presenting for lumbar spine MRI for back pain. Therefore, it should be included in the routine protocol for MR imaging of lumbar spine.
Back pain; MRI; spine; STIR
The purpose of this study was to evaluate the reliability of different anatomic reference planes in determination of Occlusal plane in dentulous and edentulous subjects with Angle’s class I and class II Maxillomandibular relationship. Eighty subjects were included in the study which was divided into four Groups based on dentition and skeletal relationship. The Group I (n = 20) and Group II (n = 20) includes young and completely dentulous subjects in Angle’s class I and class II relationship, respectively. The Group III (n = 20) and Group IV (n = 20) subjects were completely edentulous in Angle’s class I and class II relationship, respectively. For all subjects, right lateral cephalograms were taken and cephalometric analysis was done and data obtained from cephalometric tracings were then statistically analyzed. The results showed no significant difference of angle between Occlusal plane and Frankfort Horizontal plane, Camper’s plane and Palatal plane when Group I and Group III, and Group II and Group IV subjects were compared. The Frankfort Horizontal plane, Camper’s plane and the Palatal plane may be used as a reliable guide to establish Occlusal plane in edentulous subjects in both Angle’s class I and class II Maxillomandibular relationships.
Complete denture; Frankfort Horizontal plane; Occlusal plane; Reference plane
Fractures of metal-ceramic restoration pose an esthetic and functional dilemma both for patient and the dentist. Intraoral repair systems eliminate the remake and removal of restoration. Many intraoral repair materials and surface treatments are available to repair intraorally fractured metal-ceramic restoration. Bond strength data of various materials and specific technique used for repair are necessary for predicting the success of a given repair system. This study evaluated the shear bond strength of three different intraoral repair systems for metal-ceramic restorations applied on exposed metal and porcelain surface. One hundred and twenty metal discs (20 mm in diameter × 0.7 mm thick) were fabricated with nickel–chromium alloy (Mealloy, Dentsply, USA). Feldspathic porcelain (Duceram, Degudent, Germany) were applied over one test surface of the discs in the thickness of 1.8 mm followed by conventional firing. The defect, which simulates clinical failures were created in 1/4th area of the metal-ceramic discs. The metal-ceramic discs samples were divided into ceramic substrate (Group I, n = 60) and metal substrate (Group II, n = 60), according to the defect location. Then, samples of ceramic substrate (Group I) and metal substrate (Group II) were subdivided into A, B according to the surface treatments (A; roughening with diamond bur and B; abraded with 50 μ Al2O3) and repaired with one of the intraoral repair systems tested (a. Ceramic repair system, Ivoclar Vivadent; b. Clearfil repair system, Kurary, c; Porcelain repair system, 3 M ESPE). All the repaired samples were stored in distilled water at 37 °C for 24 h. After thermocycling at 6–60° C, all the samples were stored at 37 °C for additional 7 days. Shear bond strength of all the samples were calculated by using Universal testing machine. The mean shear bond strength values for the group I (A/B) were as follows: Ceramic repair system (9.47 ± 1.41/14.03 ± 2.54 MPa), Clearfil repair system (14.03 ± 2.32/14.64 ± 2.28 MPa), and Porcelain repair system (14.41 ± 3.96/14.86 ± 3.10 MPa). The mean shear bond strength values for the group II (A/B) were as follows: Ceramic repair system (9.42 ± 1.44/18.61 ± 2.60 MPa), Clearfil repair system (14.44 ± 3.23/14.98 ± 2.73 MPa), and Porcelain repair system (11.86 ± 2.24/13.24 ± 2.72 MPa). Air abrasion with 50 μm aluminum oxide particles is the preferred surface treatment. Porcelain repair system showed the highest shear bond with air abrasion for ceramic substrate and for metal substrate Ceramic repair system showed the highest bond strength with air abrasion as a surface treatment. This study suggest that the three repair systems tested are adequate for intraoral chairside repair of metal-ceramic restoration when air abrasion is used for surface treatment of the substrate (Ceramic repair system, Ivoclar Vivadent, Germany; Clearfil repair system, Kurary, Japan; Porcelain repair system, 3M ESPE, Germany).
Repair of metal-ceramic restoration; Intraoral repair; Chair side repair of metal-ceramic restoration
Changes in lifestyle lead to insulin resistance (IR) in females ultimately predisposing them towards infertility. In addition, cadmium (Cd), an environmental endocrine disruptor, is reported for detrimental effects on granulosa cells, thus leading to ovarian dysfunction. A combination of these factors, lifestyle and environment, seems to play a role in etiology of idiopathic infertility that accounts for 50% amongst the total infertility cases. To address this issue, we made an attempt to investigate the extent of Cd impact on insulin-resistant (IR) granulosa cells. We exposed adult female Charles Foster rats to dexamethasone and confirmed IR condition by fasting insulin resistance index (FIRI). On treatment of IR rats with Cd, the preliminary studies demonstrated prolonged estrous cyclicity, decrease in serum estradiol concentrations, abnormal histology of ovary, and increased granulosa cell death. Further gene and protein expression studies of steroidogenic acute regulatory (StAR) protein, 17β-hydroxysteroid dehydrogenase (17β-HSD), and cytochrome P450 aromatase (CYP19A1) were performed. Protein expression studies demonstrated significant decrease in treated groups when compared with control. Study revealed that, in spite of the molecular parameters being affected at varied level, overall ovarian physiology is maximally affected in IR and Cd coexposed group, thus mimicking the condition similar to those prevailing in infertile females.
Pluripotency and stemness is believed to be associated with high Oct-3/4, Nanog, and Sox-2 (ONS) expression. Similar to embryonic stem cells (ESCs), high ONS expression eventually became the measure of pluripotency in any cell. The threshold expression of ONS genes that underscores pluripotency, stemness, and differentiation potential is still unclear. Therefore, we raised a question as to whether pluripotency and stemness is a function of basal ONS gene expression. To prove this, we carried out a comparative study between basal ONS expressing NIH3T3 cells with pluripotent mouse bone marrow mesenchymal stem cells (mBMSC) and mouse ESC. Our studies on cellular, molecular, and immunological biomarkers between NIH3T3 and mBMSC demonstrated stemness property of undifferentiated NIH3T3 cells that was similar to mBMSC and somewhat close to ESC as well. In vivo teratoma formation with all three germ layer derivatives strengthen the fact that these cells in spite of basal ONS gene expression can differentiate into cells of multiple lineages without any genetic modification. Conclusively, our novel findings suggested that the phenomenon of pluripotency which imparts ability for multilineage cell differentiation is not necessarily a function of high ONS gene expression.
We report the first proof-of-principle demonstration of photoacoustic imaging using a contrast agent composed of a plant virus protein shell, which encapsulates indocyanine green (ICG), the only FDA-approved near infrared chromophore. These nano-constructs can provide higher photoacoustic signals than blood in tissue phantoms, and display superior photostability compared to non-encapsulated ICG. Our preliminary results suggest that the constructs do not elicit an acute immunogenic response in healthy mice.
The present study was undertaken to scientifically validate the antidiabetic activity of aqueous fruit extract of Trichosanthes dioica Roxb. (Family: Cucurbitaceae) which has been traditionally used for managing diabetes mellitus. This plant commonly known as “Sespadula” in English has not been explored scientifically so far for its glycemic potential except by our research group. The study was conducted with variable doses on normal, mild, and severe diabetics models, and several biochemical parameters including blood glucose level (BGL) were assessed. Maximum fall in BGL of 23.8% in normal rats and of 31.3% in mild diabetic rats was observed during their fasting blood glucose (FBG) and glucose tolerance test (GTT) with the dose of 1000 mg kg−1. In severely diabetic animals after 4 weeks treatment with FBG, postprandial glucose, total cholesterol, and triglyceride levels were reduced by 28.7, 30.7, 57.2, and 18.5%, whereas high density lipoprotein, total protein, hemoglobin, and body weight were increased by 33.0, 36.7, 15.7 and 16.7%, respectively. Moreover, urine sugar was reduced from +4 to +1. Thus, the study scientifically validates the traditional use of T. diocia in diabetes management and could be developed as an effective oral agent for treating diabetes mellitus and complications associated with it.
Benign Prostate hyperplasia (BPH) and prostate cancer (PCa) are the most common prostatic disorders affecting elderly men. Multiple factors including hormonal imbalance, disruption of cell proliferation, apoptosis, chronic inflammation, and aging are thought to be responsible for the pathophysiology of these diseases. Both BPH and PCa are considered to be arisen from aberrant proliferation of prostate stem cells. Recent studies on BPH and PCa have provided significant evidence for the origin of these diseases from stem cells that share characteristics with normal prostate stem cells. Aberrant changes in prostate stem cell regulatory factors may contribute to the development of BPH or PCa. Understanding these regulatory factors may provide insight into the mechanisms that convert quiescent adult prostate cells into proliferating compartments and lead to BPH or carcinoma. Ultimately, the knowledge of the unique prostate stem or stem-like cells in the pathogenesis and development of hyperplasia will facilitate the development of new therapeutic targets for BPH and PCa. In this review, we address recent progress towards understanding the putative role and complexities of stem cells in the development of BPH and PCa.
In this study, we investigated the effects of membrane cholesterol content on the mechanical properties of cell membranes by using optical tweezers. We pulled membrane tethers from human embryonic kidney cells using single and multi-speed protocols, and obtained time-resolved tether forces. We quantified various mechanical characteristics including the tether equilibrium force, bending modulus, effective membrane viscosity, and plasma membrane-cytoskeleton adhesion energy, and correlated them to the membrane cholesterol level. Decreases in cholesterol concentration were associated with increases in the tether equilibrium force, tether stiffness, and adhesion energy. Tether diameter and effective viscosity increased with increasing cholesterol levels. Disruption of cytoskeletal F-actin significantly changed the tether diameters in both non-cholesterol and cholesterol-manipulated cells, while the effective membrane viscosity was unaffected by F-actin disruption. The findings are relevant to inner ear function where cochlear amplification is altered by changes in membrane cholesterol content.
Near-infrared nanoconstructs present a potentially effective platform for site-specific and deep tissue optical imaging and phototherapy. We have engineered a polymeric nanocapsule composed of polyallylamine hydrochloride (PAH) chains cross-linked with sodium phosphate and doped with indocyanine green (ICG) toward such endeavors. The ICG-doped nanocapsules were coated covalently with polyethylene glycol (5000 daltons) through reductive amination. We administrated the constructs by tail vein injection to healthy mice. To characterize the biodistribution of the constructs, we performed in vivo quantitative fluorescence imaging and subsequently analyzed the various extracted organs. Our results suggest that encapsulation of ICG in these PEGylated constructs is an effective approach to prolong the circulation time of ICG and delay its hepatic accumulation. Increased bioavailability of ICG, due to encapsulation, offers the potential of extending the clinical applications of ICG, which are currently limited due to rapid elimination of ICG from the vasculature. Our results also indicate that PAH and ICG-doped nanocapsules (ICG-NCs) are not cytotoxic at the levels used in this study.
cancer; fluorescent imaging; nanoprobes; near infrared; pharmacokinetics; phototherapy; vascular imaging
The aim of this study was to compare color stability and surface topography of three different feldspathic porcelains both qualitatively and quantitatively after exposure to routinely consumed beverages over different time periods using a Spectrophotometer, Stereomicroscope and Surface roughness tester, respectively.
Materials and Methods:
A total of 90 plastic discs were casted to obtain metal dies for three different newer ceramic applications each on thirty samples. The color and surface roughness of these samples were measured using stereomicroscope and surface roughness tester following which they were kept in different test solutions for different durations and revaluated for color changes and surface roughness in the similar manner.
Results and Conclusion:
Among all the five test solutions, Coffee showed the maximum staining of the ceramic whereas maximum surface roughness was shown by the Duceram Kiss (1.48 μm) by Orange Juice which could be due to its high titratable acidity.
Color stability; feldspathic porcelains; spectrophotometer
The genome of retroviruses, including HIV-1, is packaged as two homologous (+) strand RNA molecules, noncovalently associated close to their 5′-end in a region called dimer linkage structure (DLS). Retroviral HIV-1 genomic RNAs dimerize through complex interactions between dimerization initiation sites (DIS) within the (5′-UTR). Dimer formation is prevented by so calledLong Distance Interaction (LDI) conformation, whereas Branched Multiple Hairpin (BMH) conformation leads to spontaneous dimerization.
Methods and Results
We evaluated the role of SL1 (DIS), PolyA Hairpin signal and a long distance U5-AUG interaction by in-vitro dimerization, conformer assay and coupled dimerization and template-switching assays using antisense PNAs. Our data suggests evidence that PNAs targeted against SL1 produced severe inhibitory effect on dimerization and template-switching processes while PNAs targeted against U5 region do not show significant effect on dimerization and template switching, while PNAs targeted against AUG region showed strong inhibition of dimerization and template switching processes.
Our results demonstrate that PNA can be used successfully as an antisense to inhibit dimerization and template switching process in HIV -1 and both of the processes are closely linked to each other. Different PNA oligomers have ability of switching between two thermodynamically stable forms. PNA targeted against DIS and SL1 switch, LDI conformer to more dimerization friendly BMH form. PNAs targeted against PolyA haipin configuration did not show a significant change in dimerization and template switching process. The PNA oligomer directed against the AUG strand of U5-AUG duplex structure also showed a significant reduction in RNA dimerization as well as template- switching efficiency.The antisense PNA oligomers can be used to regulate the shift in the LDI/BMH equilibrium.
Intradural lumbar disc herniation (ILDH) is uncommon pathology. In present report, authors present a case of ILDH associated with dorsal herniation of the cauda equina rootlets in a 30-year-old male laborer who had chronic backache since last two years. To the best of our knowledge we are reporting this for first time. Report demonstrates the natural course of ILDH.
Intradural disc herniation; Duroplasty; Herniated cauda equina rootlets
Bacterial endospores are some of the most resilient forms of life known to us, with their persistent survival capability resulting from a complex and effective structural organization. The outer membrane of endospores is surrounded by the densely packed endospore coat and exosporium, containing amyloid or amyloid-like proteins. In fact, it is the impenetrable composition of the endospore coat and the exosporium that makes staining methodologies for endospore detection complex and challenging. Therefore, a plausible strategy for facile and expedient staining would be to target components of the protective surface layers of the endospores. Instead of targeting endogenous markers encapsulated in the spores, here we demonstrated staining of these dormant life entities that targets the amyloid domains, i.e., the very surface components that make the coats of these species impenetrable. Using an amyloid staining dye, thioflavin T (ThT), we examined this strategy. A short incubation of bacillus endospore suspensions with ThT, under ambient conditions, resulted in (i) an enhancement of the fluorescence of ThT and (ii) the accumulation of ThT in the endospores, affording fluorescence images with excellent contrast ratios. Fluorescence images revealed that ThT tends to accumulate in the surface regions of the endospores. The observed fluorescence enhancement and dye accumulation, coupled with the sensitivity of emission techniques, provide an effective and rapid means of staining endospores without the inconvenience of pre- or posttreatment of samples.
The fundamental objective in restoring a congenital as well as acquired defect of eye with an ocular prosthesis is to enable the patient to cope better with the difficult process of rehabilitation after an enucleation or evisceration. A cosmetically acceptable prosthesis is that reproduces the color, form and orientation of iris and allows the patient to return to accustomed lifestyle. A sequence of steps for construction of custom-made ocular prostheses is outlined in this case report using the advantages of digital imaging technique.
Custom-made ocular prosthesis; Photographic iris; Digital imaging
Designing biomaterial scaffolds remains a major challenge in tissue engineering. Key to this challenge is improved understanding of the relationships between the scaffold properties and its degradation kinetics, as well as the cell interactions and the promotion of new matrix deposition. Here we present the use of non-linear spectroscopic imaging as a non-invasive method to characterize not only morphological, but also structural aspects of silkworm silk fibroin-based biomaterials, relying entirely on endogenous optical contrast. We demonstrate that two photon excited fluorescence and second harmonic generation are sensitive to the hydration, overall β sheet content and molecular orientation of the sample. Thus, the functional content and high resolution afforded by these non-invasive approaches offer promise for identifying important connections between biomaterial design and functional engineered tissue development. The strategies described also have broader implications for understanding and tracking the remodeling of degradable biomaterials under dynamic conditions both in vitro and in vivo.
Silk; Fibroin; Non-linear Microscopy; Two-photon excited fluorescence; Second Harmonic Generation; Spectral Analysis; Non-invasive characterization
Current approaches to study biomaterial mineralization are invasive and prevent dynamic characterization of this process within the same sample. Polarized light scattering spectroscopy (LSS) may offer a non-invasive alternative for assessing the levels of mineraliazation as well as some aspects of the organization of the mineral deposits. Specifically, we used LSS to characterize the formation of hydroxyapatite deposits on three types of silk films (water-annealed, methanol-treated and poly aspartic acid (PAA)-mixed) following 1, 3, 5 and 7 cycles of mineralization. We found that the total light scattering intensity provided a quantitative measure of the degree of mineralization as confirmed by thermal gravimetric analysis (TGA). The PAA-mixed silk films yielded the highest level of mineral deposition and the water-annealed ones the least, consistent with the β sheet content of the films prior to the onset of mineralization. The wavelength dependence of the singly backscattered light was consistent with a self-affine fractal morphology of the deposited films within scales in the range of 150 to 300 nm; this was confirmed by Fourier analysis of scanning electron microscopy (SEM) images of the corresponding films. The deposits of minerals in the water-annealed films were predominantly flake-like, with positively correlated density fluctuations (Hurst parameter, H>0.5), whereas methanol-treated and PAA-mixed silk films resulted in densely-packed, bulk mineral deposits with negatively correlated density fluctuations (H<0.5). Therefore, LSS could serve as a valuable tool for understanding the role of biomaterial properties in mineral formation, and, ultimately, for optimizing biomaterial designs that yield mineral deposits with the desired organization.
Silk; Fibroin; Tissue engineering; Polarization; Light scattering