Development and assessment of an ex vivo imaging protocol for dental-dedicated MRI systems: A pilot study

In synovial fluid, hyaluronic acid (HA) is an essential component for the lubrication of joints, thus preventing friction. The relationship between HA and joint friction is not unambiguously established yet. In the present study, the effect of the application of HA on the frictional coefficient in the temporomandibular joint was evaluated. After measuring the frictional coefficient in intact porcine joints (n = 10), the subsequent effect of phosphate-buffered saline (PBS) washing and gauze scouring and the application of HA was examined. Compared with the intact joint, the frictional coefficient was significantly larger after PBS washing and gauze scouring. Subsequent application of HA resulted in a significant decrease (50-75%) of the frictional coefficient. However, it did not recover to the same value as in the intact joints. Observations by scanning electron microscopy showed that after PBS washing, the amorphous layer of the articular cartilage was still intact, whereas it was partially collapsed after gauze scouring. In conclusion, the addition of HA did reduce the coefficient of friction under the experimental conditions in this study; the relevance to the clinical condition and the duration of the treatment effect in vivo require further investigation.

Aim: This study aims to analyze the Bio efficacy of Calcium hydroxide incorporated silver nanoparticles synthesized from Acacia nilotica as intracanal medicaments. Background: The primary objective of any endodontic treatment is the removal of the infectious tissue from the root canal which also involves the removal of debris formed during the process of removal and to prevent the reinfection. This majorly involves killing if microbes, the source of infection. This study utilizes the properties of Calcium hydroxide incorporated silver nanoparticles synthesized from Acacia nilotica as intracanal medicament in endodontic treatment for the complete removal of microbes and prevention of recurrences. Materials and method: 1% Acacia nilotica is made and kept in stirring condition for complete dissolution. Filtration was done using whatman no:1 filter paper and stored at 4°C.1 milli molar of silver nitrate was prepared and added to the filtered extract drop by drop under color change observed and the time period is noted. Centrifuge was done at 6000 rpm for 10 mins at room temperature at 4°C.Crystal violet assay was done.96 well plate was taken.100 ml of BHI medium was poured in 7 wells .1% CaOH in serial dilution was poured upto 6 wells.50 ml of enterococcus faecalis of OD600=0.1 incubated at 37 °C for 48 hours. Phosphate buffer saline (PBS) wash was done.100μL of 0.2% of crystal violet irrespective of control was added and incubated for 20 to 30 mins. Again PBS wash was done to remove excess and unbound stains.125 μL of 30% glacial acetic acid is mixed in all wells and ELISA plate reader was used to measure the observents at 570 nm. Results: AgNps synthesized from Acacia nilotica incorporated Ca(OH)₂ has a high level of biofilm inhibition against the bacteria of about more than 50% at a concentration of 0.25% whereas the normal Ca(OH)₂ showed more than 50% biofilm inhibition at 1% and showed only inhibition of 40.86% at 0.25% concentration. Conclusion: the study showed silver nanoparticles synthesized from Acacia nilotica incorporated Ca(OH)₂ had better action as intracanal medicament than Calcium hydroxide

Comparison of phytochemical profiles, antioxidant and cellular antioxidant activities of different varieties of blueberry (Vaccinium spp.)

Objective To evaluate the effects of different reconstruction settings of knowledge-based iterative reconstruction technique (IMR) on noise reduction and image quality in hepatic contrast-enhanced CT(CECT) at different radiation dose levels. Methods Patients who underwent hepatic CECT because of their diseases were enrolled in this prospective study. According to random number table, patients were randomly assigned to two groups (standard dose group, SD-group, n=29; low dose group, LD-group, n=25).All patients underwent both non-enhanced CT and triphasic CECT scan including hepatic arterial phase (HAP), portal-venous phase (PVP) and delayed phase. PVP images of SD-group were acquired with tube voltage of 120 kVp and tube current-time products of 250 mAs, and 80 kVp and 500 mAs for LD-group. PVP images were reconstructed with both filtered back projection (FBP) and IMR techniques. Settings applied in IMR reconstructions consisted of body routine (BR) and body soft tissue (BST) with three levels (L1 to L3), and image series included: L1BR, L2BR, L3BR, L1BST, L2BST and L3BST. Subjective assessment of image quality including low contrast detectability (LCD), image distortion (ID) and diagnostic confidence (DC) as well as objective image quality including image noise, signal to noise ratio (SNR) and contrast to noise ratio (CNR) were compared between groups. Effective radiation dose was recorded. Objective image quality and subjective image quality were compared by using Friedman H test and ANOVA, respectively. In addition, Student t test was used to compare effective radiation doses between groups. Results In SD-group, IMR settings of L2BST, L3BST scored highest in LCD with median score of 4; L3BST and L2BST scored lowest in ID with median score of 3, while FBP and L1BR scored highest with median score of 4; L1BR, L2BR and L1BST scored highest in DC with median score of 4, while L3BST scored lowest with median score of 3. In LD-group, the distribution of all reconstruction settings scores in LCD and ID were similar to those in SD-group; however, L1BST scored highest with median score of 4, and FBP scored lowest with median score of 2 in DC. There were statistical differences in LCD, ID and DC among all the reconstruction settings for both groups (P<0.05). In terms of FBP technique, the image noise, SNR and CNR were (9.8±2.0)HU, 13.3±3.3 and 6.0±1.9 in SD-group, and (16.2±4.1)HU, 12.9±3.3 and 6.6± 2.3 in LD-group, respectively. In SD-group, image noise exhibited a trend of decrease from L1BR to L3BST[from (4.5±0.9) HU to (3.2±0.2)HU], while SNR, CNR trended to increase (SNR: from 21.4±1.4 to 24.6±4.6; CNR: from 9.4±2.2 to 10.9±2.9, respectively). The image noise, SNR and CNR in LD-group showed the same trends as SD-group [image noise: from (5.8±0.2)HU to (3.9±0.6)HU; SNR: from 26.3±4.5 to 33.1±6.9; CNR: from 13.6±4.0 to 17.4±5.1, respectively]. In both groups, there were statistically differences in image noise, SNR and CNR among all IMR settings and FBP (F values were 15.50 to 131.39, P<0.01). The effective radiation dose of the LD-group was significantly lower than that of the SD-group [(4.19±0.22) versus (7.32±0.58) mSv, t=15.27, P<0.01). Conclusions IMR can reduce image noise and improve image quality in hepatic CECT at both standard and low dose levels. L1BR, L2BR and L1BST are the most optimized reconstruction settings for the standard dose protocol, while L1BST performs best for the low dose protocol. Key words: Liver; Tomography, X-ray computed; Radiation dosage

Antimicrobial wound lavages containing chlorhexidine gluconate (CHG) or povidone-iodine (PI) are commonly used for infection prophylaxis in arthroplasty surgery, including shoulder arthroplasty. However, the potential toxicity of these solutions to the surrounding native tissue, including rotator cuff tendons, is not well understood. The purpose of this study was to investigate the ex vivo effects of a 1-minute 0.05% CHG exposure and a 3-minute 0.35% PI exposure on the viability and biochemical content of rotator cuff tendon explants. Infraspinatus tendons (n = 6 per group) were isolated from the shoulders of Nubian goats. Tendons were submerged in 0.05% CHG in sterile water for 1 minute or 0.35% PI for 3 minutes per manufacturer guidelines, followed by phosphate-buffered saline (PBS) wash and culture in tissue medium. Control tendon specimens were bathed in PBS solution for 1 minute, followed by PBS wash and culture in tissue medium. Seven days after exposure, isolates from both the tendinous and enthesis portions were analyzed for tenocyte viability, biochemical content (collagen and glycosaminoglycan [GAG]), and matrix histology. Within the tendinous portion of the tendon, CHG and PI exposure led to reduced mean tenocyte viability compared to controls (4.35% ± 4.64% viability in the CHG group, P = .045; 14.4% ± 20.3% viability for the PI group, P = .097; vs. 35.4% ± 28.4% viability in the control group). Within the enthesis, CHG and PI exposure led to smaller and nonsignificant decreased mean tenocyte viability compared to controls (29.5% ± 22.9%, P = .336 for CHG; 37.5% ± 29.4%, P = .327 for PI). There were no significant differences in mean collagen content/wet weight or GAG/wet weight among groups within the tendinous and enthesis isolates, nor were there any appreciable differences on histology in collagen and GAG distribution among groups. Brief 0.05% CHG and 0.35% PI exposures to rotator cuff tendon may lead to reduced tenocyte viability, with the tendinous portion seemingly more susceptible than the enthesis. Additionally, at these concentrations tested, CHG appears to be more toxic than PI. These findings raise concern for the potential cytotoxic effects of low-dose CHG and PI on native tendon tissue, even after brief exposures commonly used for antimicrobial prophylaxis in shoulder arthroplasty.

Optimization of recombinant adeno-associated virus (rAAV) production has important clinical implications, as manufacturing is one of the major challenges for rAAV gene therapy. In this study, we optimized upstream and downstream processing of the rAAV production platform created by an earlier design-of-experiment approach. Our results showed that adding peptones (yeastolate, Trypton N1 or both) increased production yield by 2.8- to 3.4-folds. For downstream processing, a variety of wash buffers for an affinity resin, POROS™ CaptureSelect™ (PCS)-AAVX, were tested for their effects on rAAV8 purity, including NaCl, MgCl2, arginine, Triton X-100, CHAPS, Tween 20, octyl β-d-1-thioglucopyranoside (OTG), and low pH. The results showed that the OTG wash significantly improved the rAAV purity to 97% and reduced endotoxins to an undetectable level (<0.5 EU/mL), while retaining the yield at 92.3% of the phosphate-buffered saline (PBS) wash. The OTG wash was successfully applied to purifications of rAAV1, rAAV2, and rAAV5 using PCS-AAVX, and rAAV9 using PCS-AAV9. rAAV8 purified with OTG wash showed comparable transduction efficiency in HEK 293T cells to the rAAV8 purified with PBS wash. The optimized rAAV production process yielded 5.5-6.0 × 1014 and 7.6 × 1014 vector genome per liter of HEK 293T cells for purified rAAV8- and rAAV5-EF1α-EGFP (enhanced green fluorescent protein), respectively. The platform described in this study is simple with high yields and purity, which will be beneficial to both research and clinical gene therapy.

Multiple sclerosis (MS) and control brain slices were repeatedly washed with phosphate buffered saline (PBS) then treated for 90 seconds with an acetic acid solution pH 2.5 with citrate-phosphate-borate buffers ranging in pH from 2.2 to 11.6. The protein concentrations of PBS washes and neutralized eluates were determined, and the eluates were assayed for Ig by competition microradioimmunoassay using rabbit anti-human F(ab’)2. Successive PBS washes reduced extracellular protein to a very low level. Equivalent quantities of protein were recovered from seven MS and six non-MS brain samples after both PBS washing and acetic acid elution. However, the amount of protein needed for 50% inhibition of 125I-IgG binding to anti-human F(ab’)2 was less than 25 μg/ml in 7 of 9 samples of MS brain tissue in contrast to 3 of 12 samples of non-MS brain. Myelin basic protein did not inhibit the competition of unlabelled IgG with 125I-IgG for anti-hiunan F(ab’)2; IgG was a more effective competitor than IgA or IgM. It was possible to elute Ig from human brain at various pHs ranging from 2.2 to 11.6. At pHs 5.2 and 8.5, the distinction between MS and normal brain as determined by elution of Ig was greatest.

Doppel Induces Degeneration of Cerebellar Purkinje Cells Independently of Bax

ABSTRACTPurposes: To identify the deposition of fine (≤2.5 μm diameter) particulate matter (PM) particles (PM2.5) on contact lens surfaces and to investigate the effects of such deposition on the oxygen permeability (OP) and refractive index (RI) of contact lenses.Methods: A total of 36 contact lenses, including rigid gas permeable (RGP) lens and soft contact lens (SCL), were investigated. RGP lens (n=12) and SCL (n=12) (experimental group) were incubated in a PM2.5 solution for 24 h, after which PM2.5-treated RGP lens (n=6) and SCL (n=6) were further washed for 1 h in phosphate-buffered saline (PBS). All lenses were examined by field emission scanning electron microscopy. OP and RI of all lenses were measured.Results: Average-sized PM2.5 particles deposited on RGP contact lens and SCL surfaces after immersion in the PM2.5 solution were 3.192 ± 1.637 and 2.158 ± 1.187/100 μm2, respectively. On RGP lens surfaces, we observed both large (≥2.5 µm diameter) and small (PM2.5) particles. PM2.5 particles were deposited in diffuse patterns, primarily along the honeycomb structural border of SCL, while no PM2.5 particles were found in the honeycomb hole of SCL surfaces. Washing in PBS removed the larger PM particles from RGP lens surfaces, but left copious amounts of PM2.5 particles. In contrast, nearly all PM particles were removed from SCL surfaces after PBS washing. OP values of RGP lens and SCL appeared to be unchanged by PM2.5 deposition. RI values increased in both RGP lens and SCL groups after PM2.5 deposition. However, these increases were not statistically significant, suggesting that PM2.5 deposition itself does not cause fluctuations in contact lens RI.Conclusions: Deposition of PM2.5 particles on contact lens surfaces varies according to lens material. PM2.5 particles deposited on SCL, but only large particles on RGP surfaces were able to be removed by washing in PBS and did not appear to alter OP and RI of either lens type.

Membrane skeletons are cytoskeletal elements that have important roles in cell development, shape, and structural integrity. Malaria parasites encode a conserved family of putative membrane skeleton proteins related to articulins. One member, IMC1a, is expressed in sporozoites and localizes to the pellicle, a unique membrane complex believed to form a scaffold onto which the ligands and glideosome are arranged to mediate parasite motility and invasion. IMC1b is a closely related structural paralogue of IMC1a, fostering speculation that it could be functionally homologous but in a different invasive life stage. Here we have generated genetically modified parasites that express IMC1b tagged with green fluorescent protein, and we show that it is targeted exclusively to the pellicle of ookinetes. We also show that IMC1b-deficient ookinetes display abnormal cell shape, reduced gliding motility, decreased mechanical strength, and reduced infectivity. These findings are consistent with a membrane skeletal role of IMC1b and provide strong experimental support for the view that membrane skeletons form an integral part of the pellicle of apicomplexan zoites and function to provide rigidity to the pellicular membrane complex. The similarities observed between the loss-of-function phenotypes of IMC1a and IMC1b show that membrane skeletons of ookinetes and sporozoites function in an overall similar way. However, the fact that ookinetes and sporozoites do not use the same IMC1 protein implies that different mechanical properties are required of their respective membrane skeletons, likely reflecting the distinct environments in which these life stages must operate.

Functional magnetic resonance imaging (fMRI) is a unique window to the brain, enabling scientists to follow changes in brain activity in response to hormones, ageing, environment, drugs of abuse and other stimuli. There are two features that make fMRI unique when compared with other imaging modalities used in behavioural neuroscience. First, it can be entirely noninvasive: each animal can serve as its own control over the natural course of its life, vital for following neuroadaptation and other developmental processes critical to understanding behaviour. Second, fMRI has the spatial and temporal resolution to observe patterns of neuronal activity across the entire brain in less than a minute. Although fMRI does not have the cellular spatial resolution of immunostaining, nor the millisecond temporal resolution of electrophysiology, synchronised changes in neuronal activity across multiple brain areas seen with functional MRI can be viewed as functional neuroanatomical circuits coordinating the thoughts, memories and emotions for particular behaviours. Thus, fMRI affords a systems approach to the study of the brain, complementing and building from other neurobiological techniques to understand how behaviour is organised across multiple brain regions. In this review, we present a general background to fMRI and the different imaging modalities that can be used in fMRI studies. Included are examples of the application of fMRI in behavioural neuroscience research, along with discussion of the advantages and disadvantages of this technology.

ATP7B Mediates Vesicular Sequestration of Copper: Insight Into Biliary Copper Excretion

Background: TSE factor is parameters that affect Signal to Noise Ratio (SNR) and Contrast to Noise Ratio (CNR). TSE factor for brain MRI examination is a long TSE factor. There are differences when using TSE factor. At the theory, the brain MRI examination is using TSE factor ≥16 while at Siloam Surabaya Hospital was using TSE factor 14. The writer ever seen some noises at brain MRI image therefore the radiographer doing modification of TSE factor. The purpose of this research are to determine the influence of modification in the TSE factor value against SNR and CNR and to define the SNR and CNR optimum from that.Methods: This research is a quantitative study with an experimental approach. This research was done by MRI Philips Achieva 1,5 T with 10 modification TSE factor (8, 10, 12, 14, 16, 18, 20, 22, 24 and 26). SNR and CNR obtained by measurement of ROI in the grey matter, white matter and CSF with the result an average signal and compared with the average standard deviation of the background image. Data was analyzed by linear regression test to know the influence of TSE factor against SNR and CNR and data was analyzed by descriptive test mean rank to obtain the optimum TSE factor value.Result: The result showed that there was the inluence of TSE factor to SNR and CNR at T2W TSE axial brain. There was a significant correlation between TSE factor with all of area SNR and CNR with coefficient correlation of SNR grey matter r=0,591, with coefficient correlation of SNR white matter r=0,604, with coefficient correlation of SNR CSF r=0,687, with coefficient correlation of CNR CSF–grey matter r=0,690, with coefficient correlation of CNR CSF-white matter r=0,658. The significant value of linear regression test is (0,000*) p value (0,05). TSE factor optimum value at T2W TSE axial brain was TSE factor value 10 for SNR with mean rank SNR 45,05 and TSE factor value 8 for CNR with mean rank CNR 35,43.Conclusion: There was the influence of TSE factor to SNR and CNR at T2W TSE axial brain. TSE factor optimum value in brain MRI T2W TSE axial is 10 to SNR and TSE factor 8 to CNR.

Single cell level detection of Escherichia coli in microfluidic device

Macroautophagy (autophagy) is a process wherein bulk cytosolic proteins and damaged organelles are sequestered and degraded via the lysosome. Alterations in autophagy-associated proteins have been shown to cause neural tube closure defects, neurodegeneration, and tumor formation. Normal lysosome function is critical for autophagy completion and when altered may lead to an accumulation of autophagic vacuoles (AVs) and caspase activation. The tumor suppressor p53 is highly expressed in neural precursor cells (NPCs) and has an important role in the regulation of both autophagy and apoptosis. We hypothesized that altered lysosome function would lead to NPC death via an interaction between autophagy- and apoptosis-associated proteins. To test our hypothesis, we utilized FGF2-expanded NPCs and the neural stem cell line, C17.2, in combination with the lysosomotropic agent chloroquine (CQ) and the vacuolar ATPase inhibitor bafilomycin A1 (Baf A1). Both CQ and Baf A1 caused concentration- and time-dependent AV accumulation, p53 phosphorylation, increased damage regulator autophagy modulator levels, caspase-3 activation, and cell death. Short hairpin RNA knockdown of Atg7, but not Beclin1, expression significantly inhibited CQ- and Baf A1-induced cell death, indicating that Atg7 is an upstream mediator of lysosome dysfunction-induced cell death. Cell death and/or caspase-3 activation was also attenuated by protein synthesis inhibition, p53 deficiency, or Bax deficiency, indicating involvement of the intrinsic apoptotic death pathway. In contrast to lysosome dysfunction, starvation-induced AV accumulation was inhibited by either Atg7 or Beclin1 knockdown, and Atg7 knockdown had no effect on starvation-induced death. These findings indicate that Atg7- and Beclin1-induced autophagy plays a cytoprotective role during starvation but that Atg7 has a unique pro-apoptotic function in response to lysosome dysfunction.