scholarly journals Determination of the critical concentration required for desmin assembly

1990 ◽  
Vol 272 (1) ◽  
pp. 139-145 ◽  
Author(s):  
R G R Chou ◽  
M H Stromer ◽  
R M Robson ◽  
T W Huiatt

The critical concentration required for filament assembly in vitro from highly purified desmin was determined by both turbidity and centrifugation assays. Assembly was done in the presence of 2 mM-Ca2+, 2 mM-Mg2+ or 150 mM-Na+ at 2, 22 and 37 degrees C. Similar values for critical concentration were obtained by both assays. As temperature increased, critical concentration decreased for each cation. The critical concentration was lowest in the presence of Ca2+ at 2, 22 and 37 degrees C, but was highest in the presence of 150 mM-Na+ at 2 degrees C. Negative staining showed that supernatants from the centrifugation assays contained protofilaments, protofibrils and short particles (less than 300 nm), but pellets contained long filaments (greater than 1 micron) with an average diameter of 10 nm. As the temperature increased, both the average diameter and average length of particles in the supernatant increased. Thermodynamic analysis indicated that hydrophobic interactions were dominant during desmin assembly, but that ionic interactions might also be involved. Our results demonstrated that the specific cation and temperature and temperature-cation interactions all are important in assembly of desmin intermediate filaments.

1985 ◽  
Vol 101 (4) ◽  
pp. 1316-1322 ◽  
Author(s):  
J S Pachter ◽  
R K Liem

In this paper we describe a 66-kD protein that co-purifies with intermediate filaments from rat optic nerve and spinal cord but can be separated further by ion-exchange chromatography. This protein is distinct from the 68-kD neurofilament subunit protein as judged by isoelectric focusing, immunoblotting, peptide mapping, and tests of polymerization competence. This protein is avidly recognized by the monoclonal anti-intermediate filament antigen antibody, previously demonstrated to recognize a common antigenic determinant in all five known classes of intermediate filaments. Also, when isolated this protein binds to various intermediate filament subunit proteins, which suggests an in vivo interaction with the intermediate filament cytoskeleton, and it appears to be axonally transported in the rat optic nerve. Because of this ability to bind to intermediate filaments in situ and in vitro we have named this protein alpha-internexin. A possible functional role for the protein in organizing filament assembly and distribution is discussed.


Molecules ◽  
2019 ◽  
Vol 24 (14) ◽  
pp. 2535 ◽  
Author(s):  
Yaqiong Zhang ◽  
Bo Wang ◽  
Yan Wu ◽  
Boyan Gao ◽  
Liangli (Lucy) Yu

Zein composite particles coated with caseinate-pectin electrostatic complexes (zein-caseinate-pectin particles) were fabricated using an electrostatic deposition and liquid-liquid dispersion method without heating treatment. Compared to zein particles coated only with caseinate, the acidic stability of zein-caseinate-pectin particles was greatly improved, and the particle aggregation was suppressed at pH 3–6, especially at pH values near the isoelectric point of caseinate (pH 4–5). Besides, desirable long-term storage stability and re-dispersibility were observed. Under different zein to curcumin (Cur) feeding ratios (10:1, 20:1, 30:1 and 40:1, w/w), the Cur-loaded zein-caseinate-pectin particles had a spherical shape with an average diameter ranging from 358.37 to 369.20 nm, a narrow size distribution (polydispersity index < 0.2) and a negative surface charge ranging from −18.87 to −19.53 mV. The relatively high encapsulation efficiencies of Cur (81.27% to 94.00%) and desirable re-dispersibility were also achieved. Fluorescence spectroscopy indicated that the encapsulated Cur interacted with carrier materials mainly through hydrophobic interactions. The in-vitro release profile showed a sustained release of Cur from zein-caseinate-pectin particles in acidic aqueous environment (pH 4) up to 24 h, without any burst effect. In addition, the encapsulation retained more ABTS•+ radical scavenging capacity of Cur during 4 weeks of storage. These results suggest that zein-caseinate-pectin particles may be used as a potential delivery system for lipophilic nutrients in acidic beverages.


1992 ◽  
Vol 118 (1) ◽  
pp. 83-93 ◽  
Author(s):  
L G Tilney ◽  
D J DeRosier ◽  
A Weber ◽  
M S Tilney

After Listeria, a bacterium, is phagocytosed by a macrophage, it dissolves the phagosomal membrane and enters the cytoplasm. The Listeria than nucleates actin filaments from its surface. These newly assembled actin filaments show unidirectional polarity with their barbed ends associated with the surface of the Listeria. Using actin concentrations below the pointed end critical concentration we find that filament elongation must be occurring by monomers adding to the barbed ends, the ends associated with the Listerial surface. If Listeria with tails are incubated in G actin under polymerizing conditions, the Listeria is translocated away from its preformed tail by the elongation of filaments attached to the Listeria. This experiment and others tell us that in vivo filament assembly must be tightly coupled to filament capping and cross-bridging so that if one process outstrips another, chaos ensues. We also show that the actin filaments in the tail are capped on their pointed ends which inhibits further elongation and/or disassembly in vitro. From these results we suggest a simple picture of how Listeria competes effectively for host cell actin. When Listeria secretes a nucleator, the host's actin subunits polymerize into a filament. Host cell machinery terminate the assembly leaving a short filament. Listeria overcomes the host control by nucleating new filaments and thus many short filaments assemble. The newest filaments push existing ones into a growing tail. Thus the competition is between nucleation of filaments caused by Listeria and the filament terminators produced by the host.


1991 ◽  
Vol 19 (2) ◽  
pp. 214-218
Author(s):  
Klaus Gjervig Jensen ◽  
Ole Andersen ◽  
Mogens Rønne

The spindle-inhibiting and aneuploidy-inducing potency of in vitro exposure of PHA-stimulated human lymphocytes to organotin compounds has been studied indirectly by quantitation of chromosomal contraction and directly by determination of the frequency of aneuploidy by chromosome counting. The effects of trimethyltin chloride (TMT), dimethyltin chloride (DMT), tributyltin chloride (TBT), dibutyltin chloride (DBT), triphenyltin chloride (TPhT), and diphenyltin chloride (DPhT) on chromosal contraction were studied at concentrations of 10-3-10 9M, by measurement of the average length of chromosome 1 from asynchronous cultures of human peripheral lymphocytes. TMT, TBT, TPhT and DPhT appear to be very strong inducers of chromosomal supercontraction, indicating that these compounds are possible spindle inhibitors, while DMT and DBT seem to be ineffective. TMT, TBT, TPhT and DPhT gave rise to a little more than 100% increase in the number of hyperdiploid cells, all the increases being statistically significant except for that induced by TBT, indicating that organotin compounds are capable of inducing aneuploidy, probably by affecting spindle function.


2015 ◽  
Vol 30 (2) ◽  
pp. 27
Author(s):  
Pablo Torres -Hernández ◽  
Paulina Cebada -Martínez ◽  
Diego A. Girón -Cruz ◽  
Germán I. Garrido -Fariña ◽  
Carlos A. Alvarez -González

A partir del desove inducido y la fertilización in vitro del huevo del mero piedrero Epinephilus labriformis se describe el desarrollo embrionario y el eleuteroembrión. La inducción a la maduración y desove se realizó con dos inyecciones de 700 UI GCH∙kg-1, se procedió a realizar el desove y fertilización in vitro. El huevo se colocó en incubadoras de 6 L, a 35 UPS y se cultivó a 23, 24, 25 y 27°C hasta la eclosión. Se realizó la descripción de los estadios de segmentación, gastrulación, nerula y hasta la eclosión del eleuteroembrión a 25ºC. Se reporta el promedio ± desviación estándar del diámetro de huevo, el diámetro de la gota de aceite, la longitud notocordial y la apertura de la boca. El huevo de E. labriformis es pelágico, con corión liso y con una sola gota de aceite en posición central. El diámetro del huevo fue 726.1 ± 18.4 μm y de la gota de aceite 135.5 ± 17 μm. En el cultivo a 25º C, la primera segmentación se observó a los 40 min, siendo la discoblástula evidente a tres horas despúes la fertilización. La nerula con vesículas ópticas, somitas bien definidos y con presencia de melanóforos a las 25 horas, eclosionando a las 27 horas posteriores a la fertilización. La longitud notocordal fue 1,327 ± 186.3 μm, diámetro de la gota de aceite de 131.3 ± 17.1 μm, longitud del saco vitelino de 526.2 ± 72.8 μm y su altura 359.6 ± 74.6. La ecuación que describe el tiempo de eclosión (Te) con respecto a la temperatura de incubación (Tinc) fue: Te=119.68–(3.5793xTinc), en el intervalo de 23 a 27°C. La descripción del desarrollo embrionario y del eleuteroembrión contribuye en el conocimiento biológico de la especie y en el desarrollo de metodologías para su reproducción controlada y cultivo larval. Embryonic development of the starry grouper Epinephelus labriformis (Jenyns, 1840) and description of its eleutheroembryo From the induced spawning and in vitro fertilization of the eggs of the starry grouper Epinephelus labriformis the embryonic development and the eleutheroembryo were described. The spawning induction was performed with two injections of 700 UI GCH∙kg-1 to carry out spawning and artificial fertilization. The viable eggs were placed in incubators of 6 L with 35 UPS and raised at 23, 24, 25, and 27 °C. A chronological description of segmentation stages, gastrulation, and nerula was made until hatching of the eleutheroembryo at 25°C. The average + standard deviation of the egg diameter and the diameter of oil drop from the eleutheroembryo were reported along with the notocordial length, yolk sac length, and diameter of the eleutheroembryo´s oil drop. The egg is pelagic, spherical, transparent, smooth chorion with only one oil drop in a central position. The average diameter of the eggs is 726.1 + 18.4 μm and 135.5 + 17 μm for the oil drop. At 25°C the first segmentation was observed after 40 minutes post fertilization and the discoblastula is evident at 3 hours post fertilization. The nerula with optical vesicles, well-defined somites, and the presence of melanophores was observed at 25 hours. Eggs hatched at 27 hours post fertilization. The notochord average length of the eleuteroembryo was 1,327 + 186.3 μm, the diameter of the oil drop is 131.3 + 17.1 μm. The length of the yolk sac is 526.2 + 72.8 μm and its height is 359.6 + 74.6 μm. The equation that describes hatching time (Te) with respect to the temperature (Tinc) was: Te = 119.68 – (3.5793 x Tinc) in an interval of 23°C to 27°C (n=8; r2=0.8631; p=0.0008). The description of the embryonic development and eleutheroembryo contribute to the biological knowledge of the species and the development of methods the controlled reproduction and larviculture of E. labriformis.


2018 ◽  
Vol 17 (1) ◽  
Author(s):  
Evelyn Del Rosario Munayco-Pantoja

Objetivo: Determinar la longitud y el diámetro radicular de los incisivos laterales superiores deciduos a través de la Tomografía ComputarizadaCone Beam. Metodología: Estudio descriptivo y transversal. Se recolectaron 20 incisivos deciduos laterales superiores recién extraídos o avulsionados de la clínica odontológica de la Universidad Nacional Mayor de San Marcos, en el año 2016. Los dientes fueron escaneados utilizando la Tomografía Cone Beam, se empleó el equipo OP300 Maxio, Orthopantomograph ®. Las imágenes resultantes fueron analizadas en el software de análisis de imagen ONDEMAND3DDENTAL. Se midió el eje axial, la longitud de la corona, raíz y la longitud total de la pieza dentaria. Para el análisis estadístico se utilizó el software Stata v14 (Stata Corporation College Station, Texas), previo al análisis los datos obtenidos fueron sometidos a la prueba de normalidad de Shapiro-Wilk, cumpliendo con esto, se usaron las pruebas paramétricas ANOVA y Tde Student.  Resultados: El 100% de los incisivos laterales superiores deciduos presentó un único canal radicular. La longitud promedio de la corona y de la raíz fue de 3.8mm y 11,86mm respectivamente. El diámetro medio del canal radicular según profundidad fue de 1.11 mm a nivelcervical, de 0.94 mm a nivel medio y de 0.74 mm a nivel apical, estos mostraron diferencias estadísticamente significativas (p<0.05). Conclusiones: La longitud promedio de los incisivos laterales superiores deciduos fue de 15,68mm y el diámetro radicular disminuyo de cervical a apical. Palabras clave: Tomografía Computarizada de Haz Cónico, Odontología pediátrica, diente primario, cavidadpulpar, anatomía. Abstract Purpose: To determine the length and root diameter of the deciduous upper lateral incisors through the Cone Beam Computed Tomography.Material and Methods: This is a descriptive and transversal study. We collected 20 deciduous upper lateral incisors recently extracted or avulsed from the dental clinic of Universidad Nacional Mayor de San Marcos in 2016. The teeth were scanned using Cone Beam Tomography, where the Orthopantomograph® OP 300 Maxio was used. The resulting images were analyzed in the ONDEMAND3DDENTAL imaging software. The axial axis, length of the crown, root and the total length of the dental piece were measured. The obtained data were subjected to the Shapiro-Wilk test for normality, where the parametric tests ANOVA and Student’s T-test were used. After that, the Stata v14 software (Stata Corporation College Station, Texas) was used for the statistical analysis. Results: 100% of the deciduous upper lateral incisors presented a single root canal. The average length of the crown and the root was 3.8mm and 11.86mm respectively. The average diameter of the root canal according to depth was 1.11 mm at the cervical level, 0.94 mm at the mid-root level and 0.74 mm at the apical level. They showed statistically significant differences (p <0.05). Conclusions: The average length of the deciduous upper lateral incisors was 15.68mm and the root diameterdecreased from cervical to apical Keywords: Cone-Beam Computed Tomography, Pediatric dentistry, primary teeth, dental pulp cavity, anatomy. 


Author(s):  
S. Sathapathy ◽  
B.S. Dhote ◽  
I. Singh ◽  
M. Mrigesh ◽  
S.K. Joshi ◽  
...  

Background: The Blue bull (Boselaphus tragocamelus) is one of the biggest antelopes in Asia and is widely distributed in both the forests and adjoining villages with enough green grass.Methods: The present study was carried out on the orbital cavity and foramina of skull of six specimens of adult Blue bull (Boselaphus tragocamelus) of either sex. The biometrical parameters were measured by scale, graduated tape and digital Vernier’s caliper.Result: The average left cranio-caudal orbital diameter was found to be 5.7±0.11 cm in female, which was significantly less (p less than 0.05) than that of males, where it was recorded as 5.3±0.08 cm. Similarly, the average left orbital depth was found to be 5.7±0.08 cm in female, which was significantly less (p less than 0.05) than that of males, where it was recorded as 5.3±0.06 cm. The average length of left orbital process of frontal bone was found to be 5.1±0.05 cm in female, which was significantly less (p less than 0.05) than that of males, where it was recorded as 5.3±0.07 cm. The average diameter of left supraorbital foramen was found to be 0.31±0.001 cm in female, which was significantly less (p less than 0.05) than that of males, where it was recorded as 0.7±0.003 cm. Similarly, the average maximum distance between two cranial palatine foramina was found to be 1.8±0.05 cm in female, which was significantly less (p less than 0.05) than that of males, where it was recorded as 3.3±0.12 cm. Conclusion: Most of the biometrical observations on different parameters of orbital cavity and different foramina of skull of Blue bull were having significantly (p less than 0.05) more values in males than females. The present gross and biometrical studies would be useful to the wild life professionals for determination of sex of this animal and solving vetero-legal cases related with this species.


2000 ◽  
Vol 74 (3) ◽  
pp. 1168-1177 ◽  
Author(s):  
Reinhold Welker ◽  
Heinrich Hohenberg ◽  
Uwe Tessmer ◽  
Carola Huckhagel ◽  
Hans-Georg Kräusslich

ABSTRACT Mature human immunodeficiency virus type 1 (HIV-1) particles contain a cone-shaped core structure consisting of the internal ribonucleoprotein complex encased in a proteinaceous shell derived from the viral capsid protein. Because of their very low stability after membrane removal, HIV-1 cores have not been purified in quantities sufficient for structural and biochemical analysis. Based on our in vitro assembly experiments, we have developed a novel method for isolation of intact mature HIV-1 cores. Concentrated virus suspensions were briefly treated with nonionic detergent and immediately centrifuged in a microcentrifuge for short periods of time. The resuspended pellet was subsequently analyzed by negative-stain and thin-section electron microscopy and by immunoelectron microscopy. Abundant cone-shaped cores as well as tubular and aberrant structures were observed. Stereo images showed that core structures preserved their three-dimensional architecture and exhibited a regular substructure. Detailed analysis of 155 cores revealed an average length of ca. 103 nm, an average diameter at the base of ca. 52 nm, and an average angle of 21.3°. There was significant variability in all parameters, indicating that HIV cores are not homogeneous. Immunoblot analysis of core preparations allowed semiquantitative estimation of the relative amounts of viral and cellular proteins inside the HIV-1 core, yielding a model for the topology of various proteins inside the virion.


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