scholarly journals Microtubule minus-end stability is dictated by the tubulin off-rate

2019 ◽  
Vol 218 (9) ◽  
pp. 2841-2853 ◽  
Author(s):  
Claire Strothman ◽  
Veronica Farmer ◽  
Göker Arpağ ◽  
Nicole Rodgers ◽  
Marija Podolski ◽  
...  

Dynamic organization of microtubule minus ends is vital for the formation and maintenance of acentrosomal microtubule arrays. In vitro, both microtubule ends switch between phases of assembly and disassembly, a behavior called dynamic instability. Although minus ends grow slower, their lifetimes are similar to those of plus ends. The mechanisms underlying these distinct dynamics remain unknown. Here, we use an in vitro reconstitution approach to investigate minus-end dynamics. We find that minus-end lifetimes are not defined by the mean size of the protective GTP-tubulin cap. Rather, we conclude that the distinct tubulin off-rate is the primary determinant of the difference between plus- and minus-end dynamics. Further, our results show that the minus-end–directed kinesin-14 HSET/KIFC1 suppresses tubulin off-rate to specifically suppress minus-end catastrophe. HSET maintains its protective minus-end activity even when challenged by a known microtubule depolymerase, kinesin-13 MCAK. Our results provide novel insight into the mechanisms of minus-end dynamics, essential for our understanding of microtubule minus-end regulation in cells.

Author(s):  
Eisha Imran ◽  
Faisal Moeen ◽  
Beenish Abbas ◽  
Bakhtawar Yaqoob ◽  
Mehreen Wajahat ◽  
...  

Abstract Objectives The study aimed to evaluate and compare various commercially available local anesthetic solutions. Materials and Methods A total of 150 commercially available local anesthetic cartridges of similar composition (2% lidocaine with epinephrine 1:100,000) were randomly collected and divided into 3 groups. The designations of groups were selected from their product names such that each group consisted of 60 cartridges. Group S (Septodont, France) Group M (Medicaine, Korea) and Group H (HD-Caine, Pakistan). The samples were divided into five sub-groups, each consisting of 10 cartridges from each group to investigate each parameter. Results The acquired data was statistically analyzed and compared (using SPSS version 12). Compositional analysis revealed a non-significant (P>0.05) difference when the three Groups were compared with standard lidocaine and epinephrine solutions. The mean pH values of samples from group S, M and H respectively fell within the range of pH values of commercially available solutions. Non-significant difference in EPT values of Group S and H was found when efficacy was compared (p = 0.3), however a significant difference (p < 0.01) was observed in contrast to Group M. Anti-bacterial activity was observed in all the group and a non-significant difference in cell viability values of Group S and M was found (p = 0.6), while the difference was significant in comparison to Group H. Conclusion Within the limitations of these investigations, it appears that the properties of different manufacturers fall within the recommended ranges as mentioned in literature and do not appear to be statistically different in the variables we have tested.


2019 ◽  
Vol 8 (1) ◽  
pp. 34-37
Author(s):  
Anant Bakshi ◽  
Raeesa Khan ◽  
Bilal Ahmed

This study was carried out to evaluate the difference in prices for dental treatments carried out privately in general dental practice within the Birmingham area. Ten different practices were chosen at random, which were spread across Birmingham, in order to get a better insight into the differences across the districts. Their prices for pre-determined dental procedures were procured off the practices’ respective websites or through telephoning. The findings of this study have shown a wide variation in prices for each dental procedure, with the greatest variation in prices between practices being £850 for dental implants. The procedures with the lowest average cost were fissure sealants at £23.14. The procedure with the highest average cost was dental implants at £2,261.11. This study also showed that as more dental treatment was required, the mean cost for the dental intervention increased, regardless of the tooth being treated.


Folia Medica ◽  
2012 ◽  
Vol 54 (2) ◽  
pp. 17-21 ◽  
Author(s):  
Georgi Tz. Prisadov ◽  
Angel P. Uchikov ◽  
Kathrin Welker ◽  
Herbert Wallimann ◽  
Krassimir A. Murdzhev ◽  
...  

ABSTRACT Peripheral pulmonary tumours are often quite difficult to diagnose and treat. Their detection brings immediately the problem of whether clinicians should just wait and observe or operate the patients. The AIM of this study was to determine if there is a direct correlation between tumour size and the risk for malignancy and whether the tumor size should be considered a risk factor for malignancy. PATIENTS AND METHODS: Between 1997 and 2009, 145 patients with peripheral pulmonary tumours of less than 3 cm in diameter underwent video-assisted thoracoscopic (VATS) resection for the purpose of histologic examination of the tumor. RESULTS: The mean age of the patients was 62.60 ± 0.95 years. The youngest patient was 17 years old and the oldest - 82. The study sample included 61 women and 84 men; the men were statistically signifi cantly more than the women (57.3% and 42.07%, respectively) (t = 2.74 , P < 0.01). The total number of patients we operated were 145 with 198 resected tumours. The diameter of the lesions ranged between 0.30 cm and 3 cm (mean 1.41 ± 0.06 cm). We found that 108 (54.55%) of the tumours were malignant, and 90 (45.45%) were benign, the difference between them failing to reach statistical signifi cance (t = 1.82, P > 0.05). The mean size of malignant lesions was statistically signifi cantly greater than that of benign tumours (1.62 ± 0.08 cm vs 1.15 ± 0.06 cm). CONCLUSION: The results of this study suggest that the bigger the diameter of the nodule, the greater the percentage share of malignant tumours, which means that the size of the tumour is an important risk factor for malignancy.


Author(s):  
Charu Tyagi

Gelatin-eudragit L100 nanoparticles of wet size range 170-563nm were prepared by two step dissolvation method and the effect of different concentrations of eudragit L100 and emulsifying agent - sodium lauryl sulphate (SLS) - on the particle size were studied. Synthesized nanoparticles were characterized by attenuated total reflectance-fourier transform infrared spectroscopy (ATRFTIR) and the mean size distribution. Insulin loading was done at a pH 7.4 and the in vitro insulin release studies of nanoparticles were carried out by simulating gastrointestinal tract condition which showed the minimal insulin release at pH 2.5 (20% in 90min) while appreciable release (40% in first 30min) at pH of 7.4. This pH responsive release pattern of the synthesized nanoparticles confers on the insulin protection from proteolytic degradation in acidic environment of stomach and upper intestinal part while enhancing bioavailability in the later part of intestine.


1989 ◽  
Vol 68 (9) ◽  
pp. 1308-1312 ◽  
Author(s):  
J. Li ◽  
R.P. Ellen

The study was designed to compare the adherence of several Bacteroides species to A. viscosus. Using 3H, we labeled 24 laboratory strains, including 13 Bacteroides species and I I fresh clinical isolates of three Bacteroides species. Their adherence to A. viscosus bound to a saliva-coated mineral surface was quantified by liquid scintillation. Adherence relative to a standard strain, B. gingivalis 2561, was compared. Among the lab bacteroides, those of B. gingivalis (eight strains) were the greatest binders (mean, 80.5 ± 12.4%). Strains of other lab bacteroides bound less well (mean, 33.4 ± 6.3%). The difference in means was statistically significant (p<0.01). The mean for B. gingivalis strains was also significantly greater than that for strains of B. intermedius (51.7 ± 6.2%). Attachment of B . gingivalis was saturable in experiments in which either input concentration or time was the independent variable, indicating that B. gingivalis cells do not accumulate in this in vitro simulation of plaque formation by binding to each other. Subculture did not seem to affect the degree of binding.


2000 ◽  
Vol 182 (19) ◽  
pp. 5539-5550 ◽  
Author(s):  
Paula S. Fritsch ◽  
Mark L. Urbanowski ◽  
George V. Stauffer

ABSTRACT Many transcription factors activate by directly interacting with RNA polymerase (RNAP). The C terminus of the RNAP α subunit (αCTD) is a common target of activators. We used both random mutagenesis and alanine scanning to identify αCTD residues that are crucial for MetR-dependent activation of metE and metH. We found that these residues localize to two distinct faces of the αCTD. The first is a complex surface consisting of residues important for α-DNA interactions, activation of both genes (residues 263, 293, and 320), and activation of either metE only (residues 260, 276, 302, 306, 309, and 322) or metH only (residues 258, 264, 290, 294, and 295). The second is a distinct cluster of residues important for metE activation only (residues 285, 289, 313, and 314). We propose that a difference in the location of the MetR binding site for activation at these two promoters accounts for the differences in the residues of α required for MetR-dependent activation. We have designed an in vitro reconstitution-purification protocol that allows us to specifically orient wild-type or mutant α subunits to either the β-associated or the β′-associated position within RNAP (comprising α2, β, β′, and ς subunits). In vitro transcriptions using oriented α RNAP indicate that a single αCTD on either the β- or the β′-associated α subunit is sufficient for MetR activation of metE, while MetR interacts preferentially with the αCTD on the β-associated α subunit at metH. We propose that the different αCTD requirements at these two promoters are due to a combination of the difference in the location of the activation site and limits on the rotational flexibility of the αCTD.


2019 ◽  
Author(s):  
Tugsan Tezil ◽  
Manish Chamoli ◽  
Che-Ping Ng ◽  
Roman P. Simon ◽  
Victoria J. Butler ◽  
...  

AbstractAging is characterized by the progressive loss of physiological function in all organisms. Remarkably, the aging process can be modulated by environmental modifications, including diet and small molecules. The natural compound nordihydroguaiaretic acid (NDGA) robustly increases lifespan in flies and mice, but its mechanism of action remains unclear. Here, we report that NDGA is an inhibitor of the epigenetic regulator p300. We find that NDGA inhibits p300 acetyltransferase activity in vitro and suppresses acetylation of a key p300 target in histones (i.e., H3K27) in cells. We use the cellular thermal shift assay to uniquely demonstrate NDGA binding to p300 in cells. Finally, in agreement with recent findings indicating that p300 is a potent blocker of autophagy, we show that NDGA treatment induces autophagy. These findings identify p300 as a novel target of NDGA and provide mechanistic insight into its role in longevity.


Author(s):  
Larissa Gomes ◽  
Renata Almeida-Pedrin ◽  
Victor Ladewig ◽  
Victor Didier ◽  
Joel Santiago ◽  
...  

O objetivo deste estudo consistiu em comparar a degradação de força de cinco cores diferentes de elásticos corrente, a fim de buscar parâmetros clínicos para o uso desses elásticos nas terapias ortodônticas. Foram avaliados 10 segmentos com 6 elos de elásticos corrente de força média da marca Morelli, das cores preto, cinza, azul, rosa e transparente, provenientes de embalagens seladas e dentro do prazo de validade. Os elásticos foram distendidos até o dobro de seu comprimento e mantidos por dispositivos simples com duas pontas de metal, simulando o uso contínuo dos elásticos por parte dos pacientes. Esses dispositivos foram submersos em saliva artificial a 370C dentro de recipientes mantidos em estufa.  As forças dos segmentos de elásticos foram mensuradas com um dinamômetro ortodôntico de precisão ao início e nos intervalos de 1, 7,14, 21 e 30 dias após a imersão, e a média de força para cada grupo e tempo foi calculada. A Análise de variância (ANOVA) a dois critérios (tempo e grupos) e o teste de Tukey foram utilizados para a análise dos dados. Em todos os testes estatísticos foi adotado um nível de significância de 5%. Todas as cores de elásticos degradaram com o tempo, sendo a porcentagem média de degradação após 1 dia de 40,4%; 7 dias 48,2%; 14 dias 56,8%; 21 dias 65% e 81% após 30 dias. Entretanto, há diferença de degradação de força entre as cores dos elásticos. Os elásticos Transparente e Azul apresentaram menor degradação da força, sendo que após 30 dias o elástico Transparente se mostrou mais efetivo, seguido do Azul, Cinza, Preto e Rosa. Mesmo com a diferença na degradação de força entre as cores dos elásticos, esses dados não apresentam relevância clínica se utilizados por até 14 dias, após esse período seria melhor a utilização do elástico Transparente.   Palavras-chave: Ortodontia. Látex. Elasticidade   Abstract The aim of this study was to compare the force degradation of five different colors of elastomeric chains, in order to achieve clinical parameters for the use of these elastics in orthodontic therapies. The sample was composed by 10 segments with 6 links (medium force) for each color of elastomeric chain (Morelli); black, gray, blue, pink and transparent, from sealed packages and within the expiration date. The elastics were stretched twice their length and maintained by simple devices with two metal tips, simulating the continuous use of the elastics by the patients. These devices were submerged in artificial saliva at 370C to simulate the oral environment. The  elastic segments strengths were measured with a precision orthodontic dynamometer at the beginning and at intervals of 1, 7, 14, 21 and 30 days after immersion, and the mean strength for each group and time was calculated. Analysis of variance (ANOVA) with two criteria (time and groups) and the Tukey test were used for data analysis. In all statistical tests, a significance level of 5% was adopted. All the  elastics colors degraded over time, with the average  degradation percentage after 1 day being 40.4%; 7 days 48.2%; 14 days 56.8%; 21 days 65% and 81% after 30 days. However, there is a difference in force degradation among the  elastics colors. The Transparent and Blue elastics showed less force degradation, and after 30 days the Transparent elastic was more effective, followed by Blue, Gray, Black and Pink. Even with the difference in force degradation among the  elastics colors, these data are not clinically relevant they are used for up to 14 days, after that period it would be better to use the Transparent elastic.   Keywords: Orthodontics. Latex. Elasticity


Author(s):  
Mahsa Mohajeri ◽  
Sara Khazaei ◽  
Fariborz Vafaee ◽  
Farnaz Firouz ◽  
Somayeh Ghorbani Gholiabad ◽  
...  

Objectives: This study aimed to compare the marginal fit of temporary restorations fabricated by the conventional chairside method, 3D printing, and milling. Materials and Methods: In this in vitro, experimental study, 14 temporary restorations were conventionally fabricated over an implant abutment and analog that had been mounted in a phantom model at the site of canine tooth, using auto-polymerizing acrylic resin and putty index. In digital manufacturing, the original model was scanned, and the final restoration was designed. Fourteen temporary restorations were milled out of polymethyl methacrylate (PMMA) blocks, and 14 others were printed by a 3D printer. Temporary crowns were placed on the abutment, and images were obtained from specific areas under a stereomicroscope at x100 magnification to measure the amount of marginal gap. Data were analyzed using one-way ANOVA and Tukey’s test (α=0.05). Results: The mean marginal gap values for the temporary crowns in the 3D printing, milling, and chairside groups were 91.40, 75.28 and 51.23 µm, respectively. The crowns that were conventionally fabricated chairside exhibited the lowest marginal gap, and the difference in this respect was significant among the three groups (P<0.05). Conclusion: Temporary crowns fabricated by the chairside method showed significantly smaller marginal gap; however, the marginal gap of all three groups was within the clinically acceptable range.


2020 ◽  
Vol 2020 ◽  
pp. 1-5
Author(s):  
Liyuan Dai ◽  
Weihua Lou ◽  
Qigen Fang ◽  
Xu Zhang

Objective. Recurrence is common after inappropriate surgical procedures for parotid pleomorphic adenoma (PA). However, there are some controversies regarding intraoperative tumor rupture and disease recurrence; therefore, our goal was to clarify this relationship by describing our experience with 128 cases of recurrent parotid PA. Methods. Patients suffering from a first recurrence of parotid PA were prospectively enrolled, and data regarding the operation, pathology, immunohistochemistry, and recurrence pattern (outside the previous surgical field vs. inside the previous surgical field) were extracted and analyzed. The recurrent lesions were divided into two groups based on the location of nodularity. Results. Thirty-five patients had recurrent disease outside the previous surgical field; there were 105 nodules with a mean size of 1.0 (range: 0.4–3.0) cm and 983 nodules with a mean size of 1.55 (range: 0.5–4.5) cm within the field, and the difference was significant ( p = 0.001 ). The mean values of Ki-67 in nodules outside of and within the previous surgical field were 4.7% (range: 2%–10%) and 2.1% (range: 1%–7%), respectively, and the difference was significant ( p < 0.001 ). In nodules outside the previous surgical field, cell-rich nodules were noted in 71.6% of cases; in nodules within the previous surgical field, cell-rich nodules were found in 30.4% of cases, and the difference was significant ( p < 0.001 ). Conclusion. Tumor rupture is not the only cause of disease recurrence, and recurrent PAs outside the previous surgical field are smaller in size, have higher Ki-67 expression, and have more cell-rich nodules than those within the surgical scar.


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