scholarly journals BIOCHEMICAL AND MORPHOLOGICAL CHARACTERIZATION OF AZUROPHIL AND SPECIFIC GRANULES OF HUMAN NEUTROPHILIC POLYMORPHONUCLEAR LEUKOCYTES

1974 ◽  
Vol 63 (1) ◽  
pp. 251-269 ◽  
Author(s):  
Ursula Bretz ◽  
Marco Baggiolini

Postnuclear supernates from homogenates of purified neutrophil polymorphonuclear leukocytes (PMNs) from human blood were fractionated by zonal sedimentation and isopycnic equilibration in sucrose gradients. The fractions were characterized biochemically by measuring protein content and the activities of eight enzymes. Selected fractions were further analyzed by electron microscopy. In both centrifugation systems, azurophil and specific granules could be resolved almost completely. Azurophil granules sediment three to four times faster than the specifics and have an average density of 1.23. They contain all the peroxidase of the cells, large portions of four lysosomal hydrolases, and about half of the total lysozyme, and therefore appear to be, in biochemical terms, very similar to the azurophil granules of rabbit PMNs. The specific granules, which have an average density of 1.19, contain the remaining half of the lysozyme but appear to be free of the other components of the azurophil granules, and of alkaline phosphatase. Isopycnic equilibration disclosed a minor lysosomal population, which strongly overlaps the specific granules, and made possible the identification of a membrane-fraction which is characterized by the presence of the thiol-sensitive acid 4-nitrophenyl phosphatase and of alkaline phosphatase.

2017 ◽  
Vol 32 (6) ◽  
pp. 813-825 ◽  
Author(s):  
Bipin Gaihre ◽  
Beata Lecka-Czernik ◽  
Ambalangodage C Jayasuriya

This study was aimed at assessing the effects of silica nanopowder incorporation into chitosan-tripolyphosphate microparticles with the ultimate goal of improving their osteogenic properties. The microparticles were prepared by simple coacervation technique and silica nanopowder was added at 0% (C), 2.5% (S1), 5% (S2) and 10% (S3) (w/w) to chitosan. We observed that this simple incorporation of silica nanopowder improved the growth and proliferation of osteoblasts along the surface of the microparticles. In addition, the composite microparticles also showed the increased expression of alkaline phosphatase and osteoblast specific genes. We observed a significant increase ( p < 0.05) in the expression of alkaline phosphatase by the cells growing on all sample groups compared to the control (C) groups at day 14. The morphological characterization of these microparticles through scanning electron microscopy showed that these microparticles were well suited to be used as the injectable scaffolds with perfectly spherical shape and size. The incorporation of silica nanopowder altered the nano-roughness of the microparticles as observed through atomic force microscopy scans with roughness values going down from C to S3. The results in this study, taken together, show the potential of chitosan-tripolyphosphate-silica nanopowder microparticles for improved bone regeneration applications.


1968 ◽  
Vol 39 (2) ◽  
pp. 286-298 ◽  
Author(s):  
Dorothy Ford Bainton ◽  
Marilyn G. Farquhar

Histochemical procedures for PMN granule enzymes were carried out on smears prepared from normal rabbit bone marrow, and the smears were examined by light microscopy. For each of the enzymes tested, azo dye and heavy metal techniques were utilized when possible. The distribution and intensity of each reaction were compared to the distribution of azurophil and specific granules in developing PMN. The distribution of peroxidase and six lysosomal enzymes (acid phosphatase, arylsulfatase, ß-galactosidase, ß-glucuronidase, esterase, and 5'-nucleotidase) corresponded to that of azurophil granules. Progranulocytes contained numerous reactive granules, and later stages contained only a few. The distribution of one enzyme, alkaline phosphatase, corresponded to that of specific granules. Reaction product first appeared in myelocytes, and later stages contained numerous reactive granules. The results of tests for lipase and thiolacetic acid esterase were negative at all developmental stages. Both types of granules stained for basic protein and arginine. It is concluded that azurophil and specific granules differ in their enzyme content. Moreover, a given enzyme appears to be restricted to one of the granules. The findings further indicate that azurophil granules are primary lysosomes, since they contain numerous lysosomal, hydrolytic enzymes, but the nature of specific granules is uncertain since, except for alkaline phosphatase, their contents remain unknown.


1973 ◽  
Vol 59 (3) ◽  
pp. 696-707 ◽  
Author(s):  
Ursula Bretz ◽  
Marco Baggiolini

The localization of alkaline phosphatase in the specific granules of rabbit polymorphonuclear leukocytes was investigated. The results obtained suggest very strongly that alkaline phosphatase is a component of the granule membrane. The enzyme remains attached to the membrane upon disruption of the granules by the use of detergents or by hypotonic shock and subsequent extraction with sodium sulfate, and can be isolated together with fragments of the granule membrane by isopycnic equilibration. Treatment of the granules with high amounts of Triton-X-100, sodium deoxycholate, or hexadecyltrimethylammonium bromide releases the enzyme in soluble form. In polymorphonuclear leukocyte homogenates, lysis of the granules is needed in order to render alkaline phosphatase fully accessible to substrates. This suggests that the catalytic site of the enzyme is exposed at the inner face of the granule membrane.


Author(s):  
B. L. Soloff ◽  
T. A. Rado

Mycobacteriophage R1 was originally isolated from a lysogenic culture of M. butyricum. The virus was propagated on a leucine-requiring derivative of M. smegmatis, 607 leu−, isolated by nitrosoguanidine mutagenesis of typestrain ATCC 607. Growth was accomplished in a minimal medium containing glycerol and glucose as carbon source and enriched by the addition of 80 μg/ ml L-leucine. Bacteria in early logarithmic growth phase were infected with virus at a multiplicity of 5, and incubated with aeration for 8 hours. The partially lysed suspension was diluted 1:10 in growth medium and incubated for a further 8 hours. This permitted stationary phase cells to re-enter logarithmic growth and resulted in complete lysis of the culture.


Planta Medica ◽  
2010 ◽  
Vol 76 (05) ◽  
Author(s):  
APPR Amarasinghe ◽  
RP Karunagoda ◽  
DSA Wijesundara

2020 ◽  
Vol 13 (11) ◽  
pp. 1
Author(s):  
A. R. B. Zanco ◽  
A. Ferreira ◽  
G. C. M. Berber ◽  
E. N. Gonzaga ◽  
D. C. C. Sabino

The different integrated production systems can directly interfere with its bacterial community. The present study aimed to assess density, bacterial diversity and the influence of dry and rainy season in different integrated and an exclusive production system. The fallow and a native forest area was assessed to. Samples were collected in 2012 March and September. The isolation were carried out into Petri dishes containing DYGS medium. The number of colony forming units (CFU) was counted after 48 hours and. The bacterial density ranged between 106 and 107 CFU g-1 soil. The crop system affected the dynamics of the bacterial community only in the rainy season. The rainy season showed greater density of total bacteria when compared to the dry period regardless of the cropping system. The dendrograms with 80 % similarity showed thirteen and fourteen groups in the rainy and dry seasons. Isolates with the capacity to solubilize phosphate in vitro were obtained from all areas in the two seasons, but this feature has been prevalent in bacteria isolated during the rainy season


2020 ◽  
Vol 20 (4) ◽  
pp. 448-454
Author(s):  
Rahmita Burhamzah ◽  
Gemini Alam ◽  
Herlina Rante

Background: Endophytic fungi live in plants’ tissue and can produce the same bioactive compounds as its host plant produces. Syzygiumpolyanthum leaves have known to be one of the antibacterial compound producers. Aim and Objective: This study aimed to characterize morphologically, microscopically, and molecularly the antibacterial-producing endophytic fungi of Syzygiumpolyanthum leaves. Methods: The isolation of endophytic fungi was done by fragment planting method on PDA medium. The antibacterial screening was performed using the antagonistic test as the first screening followed by the disc diffusion test method. The morphological characterization was based on isolate’s mycelia color, growth pattern, margin, and surface texture of the colony, while the microscopic characterization was based on its hyphae characteristics. The molecular characterization of the isolate was done by nitrogen base sequence analysis method on nucleotide constituent of ITS rDNA genes of the isolate. Results: The results found that isolate DF1 has antibacterial activity against E.coli, S.aureus, P.acne, and P.aeruginosa, with the greatest inhibition at 10% concentration of broth fermentation extract on S.aureus with a diameter of inhibition of 13.77 mm. Conclusion: Based on macroscopic, microscopic, and molecular characterization, DF1 isolate is similar to Ceriporialacerate.


Membranes ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 303
Author(s):  
Rokayya Sami ◽  
Schahrazad Soltane ◽  
Mahmoud Helal

In the current work, the characterization of novel chitosan/silica nanoparticle/nisin films with the addition of nisin as an antimicrobial technique for blueberry preservation during storage is investigated. Chitosan/Silica Nanoparticle/N (CH-SN-N) films presented a stable suspension as the surface loads (45.9 mV) and the distribution was considered broad (0.62). The result shows that the pH value was increased gradually with the addition of nisin to 4.12, while the turbidity was the highest at 0.39. The content of the insoluble matter and contact angle were the highest for the Chitosan/Silica Nanoparticle (CH-SN) film at 5.68%. The use of nano-materials in chitosan films decreased the material ductility, reduced the tensile strength and elongation-at-break of the membrane. The coated blueberries with Chitosan/Silica Nanoparticle/N films reported the lowest microbial contamination counts at 2.82 log CFU/g followed by Chitosan/Silica Nanoparticle at 3.73 and 3.58 log CFU/g for the aerobic bacteria, molds, and yeasts population, respectively. It was observed that (CH) film extracted 94 regions with an average size of 449.10, at the same time (CH-SN) film extracted 169 regions with an average size of 130.53. The (CH-SN-N) film presented the best result at 5.19%. It could be observed that the size of the total region of the fruit for the (CH) case was the smallest (1663 pixels), which implied that the fruit lost moisture content. As a conclusion, (CH-SN-N) film is recommended for blueberry preservation to prolong the shelf-life during storage.


Author(s):  
N R Gazizova ◽  
A G Mannapov ◽  
V N Sattarov ◽  
V G Semenov ◽  
A I Skvortsov ◽  
...  

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