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The aim of the presented paper was to compare the Falck-Hillarp histofluoresce method for monoamines with its cryostat modification and the glyoxylic acid SPG method. The sensitivity and specificity of these methods and also the occurence of unspecific fluorescence is discussed.
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Two major forms of native sheep plasminogen (SPg-a) have been isolated from plasma by affinity chromatography. These forms differ in molecular weight, charge characteristics, affinity for epsilon-aminocaproic acid (epsilon-Ahx), and carbohydrate content. Upon treatment of SPg-a with plasmin, lower molecular weight plasminogens can be isolated. A plasminogen (SPg-b) of molecular weight approximately 8,000 less than native plasminogen is rapidly produced when either major plasminogen form is treated with plasmin. The molecular weight differences found in the major SPg-a forms are retained in the SPg-b forms, derived from each SPg-a. Upon protracted treatment of either major form of SPg-a or SPg-b with plasmin, a plasminogen (SPg-c) or molecular weight approximately 32,000 less than SPg-b is produced. A single peptide (P) is also produced in this step. The SPg-c species produced from each original SPg-a major form possess essentially the same molecular weights and carbohydrate compositions; but the P cleaved retains the molecular weight and carbohydrate differences found in each major SPg-a or SPg-b form. A large decrease in the S20,w of SPg-a is observed upon the binding of epsilon-Ahx to this protein. A much smaller alteration in the S20,w of SPg-b and SPg-c is observed upon binding of epsilon-Ahx to these proteins.
The dynamics of acute mitral regurgitation were studied in six open-chest dogs in whom a portion of the anterior leaflet was excised. Phasic mitral and aortic flows were measured electromagnetically and left ventricular filling volume, regurgitant volume (RV) and forward stroke volume (SV) were calculated. The systolic pressure gradient (SPG) between the left ventricle (LV) and left atrium (LA) was obtained from high-fidelity pressure transducers. The effective mitral regurgitant orifice area (MRA) was calculated from the hydraulic equation of Gorlin. Volume infusion resulted in significant increases in both left atrial and left ventricular pressures; thus, the SPG was unchanged and the increase in RV was due primarily to the increase in MRA. Angiotensin infused to raise arterial pressure resulted in greater increments in left ventricular than left atrial pressure, so that SPG rose significantly. The increase in RV was due to increases in both MRA and SPG. Norepinephrine infusion increased systolic left ventricular pressure and SPG, while left ventricular end-diastolic pressure and left atrial pressure diminished. Despite a significant increase in SPG, RV did not increase, due to a substantial decrease in MRA. Thus, angiotensin and volume infusion induced a substantial increase in regurgitation due to the increase in MRA, while augmentation of contractility after norepinephrine infusion resulted in a decrease in regurgitation through reduction of MRA. These findings support the clinical view that maintaining a small LV with sustained myocardial contractility will reduce mitral regurgitation. Alternatively, left ventricular dilatation can enhance mitral regurgitation by increasing the effective regurgitant orifice independent of SPG.
Effects of schizophyllan (SPG) and several antibiotics in combination against experimental Pseudomonas aeruginosa infection were examined. (1) Whereas the increase in protective effect of SPG was not observed in combination with carbenicillin or sulbenicillin, remarkable effects could be recognized in combination with dibekacin or gentamicin (GM). The combined therapy with SPG and GM proved to be most excellent. (2) The time course for changes of viable number of Ps. aeruginosa in various organs of mice revealed that the multiplication of the infected bacteria was more markedly inhibited in mice treated with SPG and GM in combination than those treated with each alone. (3) The increase in bactericidal activity of the peritoneal exudate cells was more remarkably demonstrated in the combined treatment with SPG and GM than with each alone, showing the rapid clearance of bacteria from the peritoneal cavity in the former. (4) The microscopic estimation demonstrated that the increase in both phagocytic index and intracellular digestion of bacteria by cultivated macrophages was larger in combination of SPG and GM than each alone. (5) The measurement of viable number of bacteria in cultivated macrophages by the plate-count method also showed that the ingestion and bactericidal activity within the cells were much higher in combined treatment with SPG and GM than each alone.
The delayed cutaneous hypersensitivity test with picryl chloride was used to demonstrate the effect of sonicated schizophyllan (SPG-S) on cellular immunity in mice. The increase of ear thickness in mice treated with SPG-S (0.5 approximately 10.0 mg/kg) 24 hours before the sensitization with picryl chloride proved to be significantly larger than that in untreated mice on testing with picryl chloride at 7 days following sensitization. Mice were intramuscularly injected with SPG-S (5.0 mg/kg) on -48, -24, 0, +24 or +48 hours of sensitization. Remarkable stimulating effect of SPG-S on the delayed hypersensitivity was recognized in all groups of mice, being somewhat lower in the group treated with SPG-S 48 hours after the sensitization. Stimulating effects of lipopolysaccharide, concanavalin A, pokeweed mitogen and SPG-S on the delayed hypersensitivity were compared. The effect of SPG-S was similar to that of concanavalin A, a nonspecific stimulant for T cells.
Many articles implicate the nasal ganglion in the production of remote symptoms and discuss treatment. Symptoms are primarily spastic, involving both visceral and voluntary muscles including muscle spasm in the neck, shoulder, and low back; asthma, hypertension, intestinal spasm; diarrhea, angina pectoris, uterine spasm; intractable hiccup, and many others. All these symptoms appear to have 2 common denominators. They are mediated by the autonomic nervous system and at least in some instances can be "psychosomatic." The sphenopalatine ganglion (SPG) is a major autonomic ganglion located superficially in the pterygopalatine fossa, with major afferent distribution to the entire nasopharynx and important connections with the trigeminal nerve, facial nerve, internal carotid artery plexus of the sympathetic nervous system and, as shown in the rat, direct connection with the anterior pituitary gland. This paper presents arguments supporting the following hypotheses: 1. The SPG probably has a crucial role in lower animals in declenching the reflex responses known collectively as the rage reaction. 2. The SPG is a major point of entry to the autonomic system exposed to pathologic influences and readily accessible for therapeutic influences and readily accessible for therapeutic intervention. 3. A wide variety of symptoms are produced or maintained by alteration in autonomic system tonus and some of these may be affected by intervention on the SPG. 4. The possible relationship of some symptoms and "psychosomatic" conditions to the autonomic nervous system and the rage reaction must be considered.20
Using two fluorescence histochemical methods, formaldehyde-induced fluorescence and sucrose-potassiumphosphate-glyoxylic acid fluorescence (SPG), we studied the effect of 5-hydroxytryptamine receptor stimulation by quipazine (2-[-piperazinyl]quinoline maleate) on monoamine fluorescence in the brain stem of rats. It was found that quipazine in a dose of 5 mg/kg i.p., after 60 min, decreased noradrenaline fluorescence intensity in noradrenergic neurons of the subcoeruleus area and diminished th density of catecholamine terminals visualized in the central part of the dorsal raphé nucleus. In the principal locus coeruleus, the intensity of fluorescence in nerve cells was not changed using either method, but with the SPG procedure, diffuse fluorescence outside cell bodies was observed after quipazine. In dorsal raphé neurons, a slight increase in 5-hydroxytryptamine fluorescence intensity was observed. The results obtained indicate that quipazine, apart from its effect on 5-hydroxytryptamine neurons, may also affect certain noradrenergic neurons.
CRISPR-Cas9 is a widely used platform for plant genome editing, but its outcomes are typically dominated by small insertions and deletions (indels). Such limited mutation profiles restrict its utility in functional studies of non-coding RNAs and regulatory elements, such as microRNAs (miRNAs), untranslated regions (UTRs), and promoter sequences, where larger sequence disruptions are often required. Here, we developed enhanced exonuclease-Cas9 platforms, termed multiple nucleotide deletion Cas9 (MND-Cas9) systems, for efficient generation of large deletions in rice. By screening four exonucleases (RecJ, T5, TREX2, and SbcB), we established MND-Cas9v1 systems based on TREX2 or SbcB that produced substantially larger deletions without reducing editing efficiency. Further optimization with an inserted DNA-binding domain (DBD) between Cas9 and exonuclease yielded MND-Cas9v2, which simultaneously enhanced efficiency and deletion size. To expand PAM compatibility, we introduced PAM-relaxed Cas9-NG and SpG variants, generating MND-Cas9-NG/SpGv2 systems with broader targeting scope and superior performance compared to their parental nucleases. Finally, we demonstrated the utility of these systems in two applications: MND-Cas9v2 efficiently knocked out the miRNA gene OsMIR530, producing larger seeds, and generated extended deletions in the 3'UTR of OsGhd2, which upregulated its expression and increased grain size. These results demonstrate that MND-Cas9 systems enable high-efficiency generation of extended deletions and facilitate functional analyses of non-coding RNAs and regulatory sequences. Overall, this work establishes a versatile and expandable exonuclease-Cas9 platform that substantially broadens the mutational spectrum and application potential of CRISPR-Cas9 for plant genome engineering.
Mouse peritoneal macrophages, but not HeLa cells, presented many characteristics of activated cells following treatment in culture with LTN or PSK. Indeed, they demonstrated increase in size, spread, endocytic and antiparasitic activities. These data lead favorably to consider that among the five polysaccharides tested under the present experimental model LTN and PSK are the most potent activators of resident mouse peritoneal macrophages in vitro. The mechanism of macrophage activation by LTN and PSK should be different from that of Di Luzio glucans and SPG since activated macrophages can be induced by the latter two compounds in experimental animals but not in vitro in cell cultures.
Karyotypes of Su-1, Su-2, KMMA-1, and KMMA-2 (producing oncornavirus C-type), and KMPG-1, SPF-2, and SPG-3 (producing HSV, a virus of EBV type) cell lines were studied in the time course. The lines were derived from hemopoietic cells of brown macaques and baboons bearing lymphomas. Some of the lines had a paradiploid-pseudodiploid karyotype, another part aneuploid type. The selectivity of structural and numerical abnormalities of chromosomes 1, 9, 10, 11, 12, 13, 15, 18, and 20 was detected indicating a certain specificity of karyotype changes upon malignization and transformation. The number of poorly and moderately differentiating chromosomes in aneuploidy was confirmed by greater variability as compared with well-differentiating chromosomes. No parallelism between the karyotype change pattern and the type of virus produced (C or EBV) could be found thus far.