scholarly journals Visual and Microscopic Evaluation of Streptomyces Developmental Mutants

Author(s):  
Jennifer A. Bennett ◽  
Garrett V. Kandell ◽  
Sean G. Kirk ◽  
Joseph R. McCormick
Author(s):  
T. M. Seed ◽  
M. H. Sanderson ◽  
D. L. Gutzeit ◽  
T. E. Fritz ◽  
D. V. Tolle ◽  
...  

The developing mammalian fetus is thought to be highly sensitive to ionizing radiation. However, dose, dose-rate relationships are not well established, especially the long term effects of protracted, low-dose exposure. A previous report (1) has indicated that bred beagle bitches exposed to daily doses of 5 to 35 R 60Co gamma rays throughout gestation can produce viable, seemingly normal offspring. Puppies irradiated in utero are distinguishable from controls only by their smaller size, dental abnormalities, and, in adulthood, by their inability to bear young.We report here our preliminary microscopic evaluation of ovarian pathology in young pups continuously irradiated throughout gestation at daily (22 h/day) dose rates of either 0.4, 1.0, 2.5, or 5.0 R/day of gamma rays from an attenuated 60Co source. Pups from non-irradiated bitches served as controls. Experimental animals were evaluated clinically and hematologically (control + 5.0 R/day pups) at regular intervals.


Author(s):  
P. M. Lowrie ◽  
W. S. Tyler

The importance of examining stained 1 to 2μ plastic sections by light microscopy has long been recognized, both for increased definition of many histologic features and for selection of specimen samples to be used in ultrastructural studies. Selection of specimens with specific orien ation relative to anatomical structures becomes of critical importance in ultrastructural investigations of organs such as the lung. The uantity of blocks necessary to locate special areas of interest by random sampling is large, however, and the method is lacking in precision. Several methods have been described for selection of specific areas for electron microscopy using light microscopic evaluation of paraffin, epoxy-infiltrated, or epoxy-embedded large blocks from which thick sections were cut. Selected areas from these thick sections were subsequently removed and re-embedded or attached to blank precasted blocks and resectioned for transmission electron microscopy (TEM).


Author(s):  
J. W. Horn ◽  
B. J. Dovey-Hartman ◽  
V. P. Meador

Osmium tetroxide (OsO4) is a universally used secondary fixative for routine transmission electron microscopic evaluation of biological specimens. Use of OsO4 results in good ultrastructural preservation and electron density but several factors, such as concentration, length of exposure, and temperature, impact overall results. Potassium ferricyanide, an additive used primarily in combination with OsO4, has mainly been used to enhance the contrast of lipids, glycogen, cell membranes, and membranous organelles. The purpose of this project was to compare the secondary fixative solutions, OsO4 vs. OsO4 with potassium ferricyanide, and secondary fixative temperature for determining which combination gives optimal ultrastructural fixation and enhanced organelle staining/contrast.Fresh rat liver samples were diced to ∼1 mm3 blocks, placed into porous processing capsules/baskets, preserved in buffered 2% formaldehyde/2.5% glutaraldehyde solution, and rinsed with 0.12 M cacodylate buffer (pH 7.2). Tissue processing capsules were separated (3 capsules/secondary fixative.solution) and secondarily fixed (table) for 90 minutes. Tissues were buffer rinsed, dehydrated with ascending concentrations of ethanol solutions, infiltrated, and embedded in epoxy resin.


Author(s):  
W.T. Gunning ◽  
J.N. Turner ◽  
K. Buttle ◽  
E.P. Calomeni ◽  
N.A. Lachant ◽  
...  

There are a variety of conditions which have been associated with prolonged bleeding times. If other etiologies including von Willebrand's disease have been ruled out, a platelet function disorder must be considered. The best, if not only, technique to make this diagnosis is the electron microscopic evaluation of whole air dried platelets. Bull first described the presence of dense granules in whole platelets in 1968 and the technique has been utilized extensively The electron dense or delta granules are easily distinguished from the larger more numerous alpha granules which are electron lucent. The significance of the dense granules is that they are known to be “storage pools” of serotonin, calcium, adenosine di- and triphosphate, and pyrophosphate. Prolonged bleeding times may be directly related to an insufficiency of these substances. The diagnosis of a storage pool deficiency is made when either the storage content of the dense granules is abnormal or their number is diminished. We observe normal platelets to have 4-6 dense granules, which agrees with the literature.


1970 ◽  
Vol 23 (02) ◽  
pp. 261-275 ◽  
Author(s):  
G Zbinden ◽  
J. N Mehrishi ◽  
S Tomlin

SummaryThe severity of platelet damage induced by hyper- and hypotonic NaCl solutions and freezing and thawing was assessed by microscopic evaluation and measuring inhibition of 5-HT uptake. The same techniques were used to quantitate the effects of aggregating agents. The positively charged macromolecules PS, Poly-L und Poly-O reduced the net negative surface charge as determined by microelectrophoresis, caused platelet aggregation and inhibited 5-HT uptake. The damaging effects of Poly-L and Poly-O were more severe and more closely related to concentration than that of PS. The negatively charged macromolecules Poly-IC and NaPS increased the anodic electrophoretic mobility. Poly-IC and heparin caused a low degree of platelet clumping and no inhibition of 5-HT uptake. NaPS produced severe platelet damage with extensive clumping and complete inhibition of 5-HT uptake. Na laurate had the same effect, but did not alter electrophoretic mobility. ADP caused concentration-dependent platelet aggregation and inhibition of 5-HT uptake. The effects of ADP and NaPS were compared in agitated and non-agitated platelet samples containing identical concentrations of the 2 compounds. Agitation was found to increase the degree of platelet clumping and to reduce 5-HT uptake.


Author(s):  
Radhika Thundikandy ◽  
Gowri Priya Chidambaranathan ◽  
Naveen Radhakrishnan ◽  
Rathinam Sr

1977 ◽  
Vol 29 (1) ◽  
pp. 55-63 ◽  
Author(s):  
R. T. M. Poulter ◽  
N. K. Honey

SUMMARYThe homothallic amoebal clones of Physarum polycephalum are of potential use in understanding the developmental genetics of this organism. Such an application requires that complementation and recombination analysis be possible between pairs of homothallic clones. This paper is a report of the formation of mixed plasmodia by pairs of homothallic amoebal clones. In order to detect such mixed plasmodia use was made of two marker genes involved in plasmodial fusion, fusA and fusB. Sporulation of a mixed plasmodium formed from two homothallic (delayed) amoebal clones yielded progeny amoebae which were genetically recombinant. It is deduced from the ratios of various genotypes in these progeny clones that the mixed plasmodium was diploid and that meiosis was associated with sporulation. There is therefore no impediment to the use of the homothallic strains for genetical analysis. The progeny amoebal clones were observed to be showing segregation for the characters homothallic (rapid) and homothallic (delayed). This observation, taken together with other related observations, suggests that the homothallic (delayed) character is produced by mutation of the homothallic (rapid) character. The rare plasmodia formed by a homothallic (delayed) amoebal clone are the result of reversion of this mutation. Amoebal clones of the homothallic (delayed) type are therefore developmental mutants unable to perform the differentiation from amoeba to plasmodium.


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