Third harmonic generation (THG) microscopy is a label‐free imaging technique that shows great potential for rapid pathology of brain tissue during brain tumor surgery. However, the interpretation of THG brain images should be quantitatively linked to images of more standard imaging techniques, which so far has been done qualitatively only. We establish here such a quantitative link between THG images of mouse brain tissue and all‐nuclei‐highlighted fluorescence images, acquired simultaneously from the same tissue area. For quantitative comparison of a substantial pair of images, we present here a segmentation workflow that is applicable for both THG and fluorescence images, with a precision of 91.3 % and 95.8 % achieved respectively. We find that the correspondence between the main features of the two imaging modalities amounts to 88.9 %, providing quantitative evidence of the interpretation of dark holes as brain cells. Moreover, 80 % bright objects in THG images overlap with nuclei highlighted in the fluorescence images, and they are 2 times smaller than the dark holes, showing that cells of different morphologies can be recognized in THG images. We expect that the described quantitative comparison is applicable to other types of brain tissue and with more specific staining experiments for cell type identification.
To shed light on the mechanism of hydrophobic control in reactions of microbial tryptophanase the direct effect of the solvent hydrophobicity on affinities of amino acid inhibitors was first examined. Values of inhibition constants (Ki) for a variety of amino acids were determined in 37.5% aqueous methanol, and no general correlation between the change of Ki, on passing from water to aqueous methanol, and amino acid hydrophobicity was found. The solvent effects on the separate stages of the external aldimine formation (KD) and deprotonation to form a quinonoid intermediate (Kq) were determined for the reactions of tryptophanase with 2-oxindolyl-
-alanine and
-alanine by stopped-flow technique. For 2-oxindolyl-
-alanine, which is a close transition-state analogue for the enzyme reaction with natural substrate, the decrease in the affinity in aqueous methanol is associated exclusively with the α-proton abstraction stage but not with the preceding formation of external aldimine. We conclude that the environment of amino acid side chains in the active site cannot be considered to be permanently hydrophobic irrespective of the bound amino acid. We suggest that complexes of tryptophanase with amino acids may exist either in a hydrophobic, presumably “closed”, conformation, where bound amino acids are isolated from the solvent, or in an accesible to solvent, “open”, conformation, depending on the structure of the bound amino acid and stage of the catalytic mechanism. For 2-oxindolyl-
-alanine the transfer from an open to a closed conformation probably accompanies deprotonation of the external aldimine. The change of the active site hydrophobicity may provide an efficient way of modulating the relative acid–base properties of the catalytic groups to ensure the movement of protons in the “correct” direction depending on the elementary stage of catalysis. 相似文献
Behavioral activity of 7-week-old German shepherd puppies was tested and the activities analyzed if they could be used for predicting police efficiency of the individual. In total 206 individuals sired by 42 sires and 44 dams were used. The activities were divided into 10 tasks in which reactions and behavior of pups were scored from 0 to 5 points. All pups were tested separately from other conspecifics. Probability that the puppy will pass the certification was tested by a logistic regression. Of the 206 puppies, 148 passed the certification while 58 failed. Some tested behavioral variables were moderately to highly correlated with one another. Therefore we applied a factor analysis. Three factors were retained accounting for 100% of the shared variance. After inspection of the rotated factor pattern matrix and its confidence intervals, it appeared that variables “Independent movement and interactions with the tester”, “Negotiating obstacles”, “Entering a room”, “Behavior toward a person”, and “Behavior in new environments” loaded on Factor 1 (“Factor for movement”), while variables “Response to distracting stimuli caused by a shovel”, “Response to a distracting noise while left alone in a room”, and “Response to loud distracting stimuli” on Factor 2 (“Factor for responding to noise”) and variables “Retrieval” and “Tug of war” on Factor 3 (“Factor for attitude to predation”). In the final logistic regression model, the probability that the puppy will pass the certification depended on the higher weight at the time of testing (, P = 0.0005), on the “Factor for attitude to predation” (, P = 0.0007), on the “Factor for responding to noise”, where the higher the score, the weaker was the response (, P = 0.0232), and on the “Factor for movement” showing an increasing probability with decreasing score (, P = 0.0219). The tests in our study seem to be a good base which might enable selection for suitable dogs as early as 7 weeks of age. The puppies having high probability to pass certification in adulthood were heavy individuals willing to chase, catch, and fetch a tennis ball, and follow a rag drawn away from them, while weakly responding to a distracting noise in various situations and showing low activity while negotiating obstacles and moving and interacting with the tester. To conclude, contrary to skeptical assumptions expressed by various authors, the specific puppy tests for police dogs provide a reliable tool for predicting future service ability of a puppy. Differences in methodology are likely to contribute to a lack of consensus among various studies. 相似文献
The effect of glycolysis remains largely elusive in acute T lymphoblastic leukemia (T-ALL). Increasing evidence has indicated that the dysregulation of miRNAs is involved in glycolysis, by targeting the genes coding glycolysis rate-limiting enzymes. In our previous studies, we found that overexpression of the ARRB1-derived miR-223 sponge repressed T-ALL progress and reduced the expression of miR-652-5p. However, little is known about miR-652-5p on T-ALL. Here, we showed that impaired miR-652-5p expression inhibited growth, promoted apoptosis of T-ALL cells in vitro and prolonged overall survival (OS) in vivo. Based on the GO enrichment of miR-652-5p target genes, we uncovered that impaired miR-652-5p decreased glycolysis, including reduced the lactate, pyruvate, ATP level and the total extracellular acidification rate (ECAR), elevated oxygen consumption rate (OCR) in T-ALL cell lines. Mechanically, miR-652-5p targeted the 3ʹUTR of Tigar mRNA and inhibited its expression. Furthermore, the alteration of glycosis level was attributed to Tigar overexpression, consistent with the effect of impaired miR-652-5p. Additionally, Tigar suppressed the expression of PFKFB3, a glycolysis rate-limiting enzyme, in vivo and in vitro. Taken together, our results demonstrate that impaired miR-652-5p/Tigar axis could repress glycolysis, thus to slow growth of T-ALL cells, which support miR-652-5p as a novel potential drug target for T-ALL therapeutics.Subject terms: Paediatric cancer, Mechanisms of disease相似文献
The effects of rodents on forest regeneration have been highlighted in many ecological studies. In 2002 and 2003, the acorns of Quercus aliena var. acuteserrata were subjected to 12 different treatments. The daily dynamics and the amount of acorns that were finally left intact, predated in situ, or removed off were examined and documented. The ratios of acorns that were infested by bugs before and after predation by rodents were carefully documented. It was found that: (1) the ratios of acorns infested by bugs before and after predation by rodents showed significant difference (P > 0.05), suggesting that rodents would not prey on acorns during the predating process if acorns had been already infested by bugs. (2) When compared with the controls, the fate of acorns could be roughly classified into four types: acorns that were simply buried or placed on black paper showed no significant differences with the controls in their response to rodents, suggesting that rodents have no sensitivity to the little change of odor resulting from burying and may be more adapted to black background. Acorns attached with strings, dyed with black ink, cut into halves, or placed on white/green/red paper were predated in situ with much lower predation rates and lasted the same duration. Compared with the controls, the acorns were removed away by rodents with the same first day rate and the removal lasted longer. The final removal rate of the acorns by rodents was increased, whereas that of predation in situ was decreased, and none were left in the spot. This may suggest that rodents in this case increased their predation risk expectation and adopted a strategy of “less predation proportion in situ”. Acorns that were burnt were not predated in situ as well as removed in the first day, but the rates were raised to the maximum in the forth and fifth day and then declined to zero. Consequently, the final ratios of predation in situ and removal considerably decreased with many acorns, nearly 50%, being left intact. It can be hypothesized that the rodents responded to the change in odor of acorns by using the strategies of “less predation proportion in situ” and “less predating activities”, thereby resulting in larger number of acorns remained intact. Acorns that were enwrapped were barely predated in situ, removed in the first day with a normal predation rate, which declined to nearly zero in the second day and then increased till it declined again from the fourth day to the final day. The final rate of predation in situ was lower than 5% without acorns remained intact, and the rate of removal of acorns showed a considerable relative increase. This suggests that when there was a change in both odor and status of acorns, rodents also adopted the “spying” and “less predation proportion in situ” strategy, Because the predation risk is critical, the sense of sight would play an important role in rodent's living. (3) Definite amounts of acorns were predated in situ, discarded, removed away, predated ex situ, lost and hoarded separately. 相似文献
Poly-N-acetyllactosamines provide backbone structures for functional modifications such as sialyl Lewis X. To understand how the biosynthesis of poly-N-acetyllactosamines is regulated, two branched oligosaccharides of the structure Gal1,4GlcNAc1, 6(Gal1,4GlcNAc1,2)-Man1,6Man-octyl 1 and 2 were synthesized in which one of the terminal galactose units was selectively radiolabeled. Hexasaccharides 1 and 2 were assembled from the chemically synthesized pentasaccharide precursors GlcNAc1,6(Gal1,4GlcNAc1,2)-Man1,6Man-octyl3 and Gal1,4GlcNAc1,6(GlcNAc1, 2) - Man1,6 Man-octyl 4 respectively, through treatment with UDP-1-[3H]-Gal and 1,4 galactosyltransferase. Compounds 1 and 2 were subsequently incubated with UDP-GlcNAc and the UDP-GlcNAc: Gal1-4Glc(NAc)1,3-N-acetylglucosaminyltransferase (i-GlcNAc transferase) resulting in a partial conversion to a mixture of heptasaccharides which were purified by HPLC. The branch selectivity of the addition of N-acetylglucosamine to compounds 1 and 2 was then characterized by endo--galactosidase digestion of the heptasaccharides, followed by isolation of the resultant pentasaccharides on C18 reverse-phase silica cartridges. Comparison of the amount of radiolabel to a control reaction lacking endo--galactosidase indicated the favored site of GlcNAc addition to be the lower 1,2-branch over the 1,6-branch by a 3:1 ratio. 相似文献
Morphological features of Monorhizochytrium globosum NBRC 112723 in seawater/seaweed culture. A single ectoplasmic net penetrated into Pyropia sp. (top left). Continuous observations of zoospore formation (from top center to right bottom); after cleavage of the multinucleate protoplasm of the zoosporangium, each uninucleate cell developed into a zoospore. See Doi and Honda in pages 188–201. Cover picture from: Article link here
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