scholarly journals SPECTROPHOTOMETRY OF THE PERIODIC ACID-SCHIFF REACTION WITH PITUITARY HORMONES IN VITRO AND IN HISTOLOGICAL SECTIONS

1962 ◽  
Vol 13 (2) ◽  
pp. 239-247 ◽  
Author(s):  
Sally B. Fand ◽  
Bo Thorell
Keyword(s):  
Periodic Acid ◽  
Periodate Oxidation ◽  
Quantitative Measure ◽  
Pituitary Hormones ◽  
Experimental Conditions ◽  
Human Pituitary ◽  
Periodic Acid Schiff ◽  
Histological Sections ◽  
Schiff Reaction

The spectral light absorption of the in vitro periodic acid-Schiff reactions of 4 purified pituitary hormones is described. The absorption spectra present a maximum between 560 and 565 mµ. The color developed conforms with Beer's law for the ranges of concentration examined. The different hormones exhibit different chromogenicity per unit of biological activity: the color produced by 1 unit of FSH is equivalent to approximately 2 of TSH, 4 of LH, and 30 of ACTH. Microspectrophotometric measurements of the PAS-positive structures in histological sections of the human pituitary give absorption curves with shapes similar to those obtained in vitro, although quantitative differences exist. It is concluded that under the proper experimental conditions microspectra of the pituitary structures might, in the future, prove to give a quantitative measure of aldehyde groups generated from glycoprotein tropins by periodate oxidation.

Effect of pepsin on partially purified pig gastric mucus and purified mucin

1988 ◽  
Vol 66 (5) ◽  
pp. 367-373 ◽  
Author(s):  
Sum P. Lee ◽  
Jane F. Nicholls ◽  
Anthony M. Roberton ◽  
Han Z. Park
Keyword(s):  
Periodic Acid ◽  
Gastric Mucin ◽  
Low Density ◽  
Gastric Mucus ◽  
Experimental Conditions ◽  
Periodic Acid Schiff ◽  
Pepsin Digestion ◽  
Cscl Density Gradient ◽  
Peptic Digestion

Partially purified native-pig gastric mucus and purified pig gastric mucin, prepared by column chromatography and caesium chloride (CsCl) density-gradient ultracentrifugation, were subjected to pepsin digestion. The products of peptic digestion were chromatographed on Sepharose CL-2B, and fractions were assayed for carbohydrate by the periodic acid – Schiff reaction. The polymeric gastric mucin in the purified mucin samples was readily degraded by pepsin. In sharp contrast, the polymeric mucin in the partially purified mucus was relatively resistant to pepsin digestion. In 45 min, pepsin degraded 40% of the polymeric mucin in the purified samples, whereas it produced no significant degradation (<10%) in the partially purified mucus samples. In partially purified gastric mucus, treated with CsCl but not fractionated by ultracentrifugation, digestion with pepsin was also slow and incomplete. This showed that differences in susceptibility between partially purified and purified preparations are not due to the chaotropic effects of CsCl. In addition, the recombination of low-density nonmucin fractions in CsCl ultracentrifugation with the mucin also resisted pepsin digestion. Finally, we have shown that the low-density fractions in mucus exhibited a strong inhibitory effect of peptic activity in vitro. We conclude that under our experimental conditions, pepsin has little effect on partially purified mucus, and our findings indicate an inhibitor of peptic digestion is present in native gastric mucus. It is likely, but unproven, that this inhibitor is a noncovalently bound lipid present in the low-density fraction.

scholarly journals Histochemical properties of cartilage proteoglycans.

1983 ◽  
Vol 31 (1) ◽  
pp. 53-61 ◽  
Author(s):  
E Ippolito ◽  
V A Pedrini ◽  
A Pedrini-Mille
Keyword(s):  
Alcian Blue ◽  
Magnesium Chloride ◽  
Periodic Acid ◽  
Keratan Sulfate ◽  
Periodic Acid Schiff ◽  
Histological Sections ◽  
Connective Tissue Matrix ◽  
Cartilage Proteoglycans ◽  
Tissue Matrix

Proteoglycan interaction with alcian blue at different concentrations of magnesium chloride was studied both in vitro and in histological sections of paraffin-embedded tissues. Our experiments indicate that a) proteoglycans with different contents of chondroitin sulfate and keratan sulfate, prepared under nondegradative conditions, are not distinguishable on the basis of the critical electrolyte concentrations at which staining is abolished; b) the state of aggregation of proteoglycans only very slightly affects the alcian blue affinity of the macromolecules at different concentrations of magnesium chloride; c) the interaction of proteoglycans with other components of the connective tissue matrix is an important factor in determining the strength of binding of alcian blue to the polyanionic macromolecules in histological sections. These factors should be considered in interpreting histochemical data obtained by staining tissue sections with alcian blue at different concentrations of magnesium chloride. Proteoglycans, like glycosaminoglycans, are only weakly periodic acid-Schiff-positive.

scholarly journals SAPONIFICATION-INDUCED INCREASE IN THE PERIODIC ACID-SCHIFF REACTION IN THE GASTROINTESTINAL TRACT MECHANISM AND DISTRIBUTION OF THE REACTIVE SUBSTANCE

1973 ◽  
Vol 21 (5) ◽  
pp. 473-482 ◽  
Author(s):  
P. E. REID ◽  
C. F. A. CULLING ◽  
W. L. DUNN
Keyword(s):  
Alternative Hypothesis ◽  
Periodic Acid ◽  
Periodate Oxidation ◽  
Periodic Acid Schiff ◽  
Initial Oxidation ◽  
Vicinal Diol ◽  
Sodium Borohydride Reduction ◽  
Vicinal Diols ◽  
Schiff Reaction ◽  
Carbonyl Function

The mechanism and distribution of the KOH-periodic acid-Schiff (PAS) effect in rat large intestine, human sigmoid colon, rabbit rectum and rabbit Brunner's glands have been investigated following the abolition of normally occurring PAS-positive vicinal diols by means of a periodate oxidation-sodium borohydride reduction sequence. The KOH-PAS effect was shown to be independent of the initial oxidation time and could be abolished by pretreatment with either methanolic-HCl or alcoholic KOH. The evidence suggests that the KOH-PAS effect is due to the removal of an o-acyl-ester substituent on a potential vicinal diol. The alternative hypothesis, that the effect is due to an ester involving the carbonyl function of a macromolecule or a lactone, is less likely. The difficulty of interpretation of histochemical results following methylation is discussed.

scholarly journals The Periodic Acid-Schiff Reaction in Neutrophil Leukocytes in Untreated and Myleran-Treated Chronic Myelocytic Leukemia

Blood ◽  
1966 ◽  
Vol 28 (4) ◽  
pp. 544-552 ◽  
Author(s):  
GÖSTA GAHRTON
Keyword(s):  
Periodic Acid ◽  
Quantitative Measure ◽  
Normal Subjects ◽  
Chronic Myelocytic Leukemia ◽  
Periodic Acid Schiff ◽  
Reactive Material ◽  
Myelocytic Leukemia ◽  
Pas Reaction ◽  
Schiff Reaction ◽  
Neutrophil Leukocytes

Abstract Microspectrophotometric quantification of the periodic acid-Schiff (PAS) reaction has been performed in about 3,000 individual mature neutrophil leukocytes from 20 normal subjects and 23 patients with chronic myelocytic leukemia (CML), of whom 10 were untreated or in relapse and 13 in partial remission or remission. The PAS reaction was taken as a quantitative measure of the cellular amount of PAS reactive material (PASMa), most probably equivalent to glycogen. The mean amount of PASMa in neutrophils from untreated and relapse cases of CML was 43 per cent less than in normal neutrophils. The frequency distribution of PASMa per cell was normal in individual cases. In clinical remission after Myleran treatment, the neutrophils contained normal amounts of PASMa.

TISSUE CULTURE STUDIES ON HUMAN PITUITARY TUMOURS: RADIOIMMUNOASSAYABLE ANTERIOR PITUITARY HORMONES IN THE CULTURE MEDIUM

1978 ◽  
Vol 88 (2) ◽  
pp. 239-249 ◽  
Author(s):  
Loren G. Lipson ◽  
Inese Z. Beitins ◽  
Paul D. Kornblith ◽  
Janet W. Mc Arthur ◽  
Henry G. Friesen ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Cushing’S Disease ◽  
Anterior Pituitary ◽  
Pituitary Hormones ◽  
Cushing's Disease ◽  
Hormone Release ◽  
Pituitary Tumours ◽  
Human Pituitary ◽  
Anterior Pituitary Hormones

ABSTRACT A tissue culture study was undertaken to determine if human non-functioning pituitary tumours secrete polypeptide anterior pituitary hormones in vitro and to study the spectrum of hormone release by functioning pituitary neoplasms. Fragments from 48 human pituitary tumours (from patients - 2 with Cushing's disease, 1 with Nelson's syndrome, 5 with amenorrhoea-galactorrhoea, 10 with acromegaly and 30 with non-functioning pituitary tumours) and three normal human anterior pituitary glands (controls) were placed in tissue culture immediately after surgery. The in vitro release of human growth hormone (HGH), prolactin (Prl), thyrotrophin (TSH), adrenocorticotrophin (ACTH), luteinizing hormone (LH) and follicle stimulating hormone (FSH) were measured by radioimmunoassays at the end of one week in culture. Clinical and pathological data were compared to hormone release patterns. In the culture media from control pituitaries the concentrations of the six hormones tested were 100 to 10 000 times greater than in peripheral blood. The medium surrounding the fragments from functioning pituitary tumours contained the following: a) Acromegaly - high levels of HGH and variable concentrations of the other hormones. b) Cushing's disease - ACTH and Prl predominantly. c) Amenorrhcea-galactorrhoea syndrome - prolactin in 4 out of 5 patients, all six polypeptides in one patient. In the media from the 30 patients diagnosed as having non-functioning pituitary tumours, 60 % of the samples contained at least one hormone at a concentration similar to that of the controls and 100 % of the samples contained detectable quantities of at least one hormone.

scholarly journals Effect of activity on performance and morphology in ischaemic rat slow muscles

1990 ◽  
Vol 152 (1) ◽  
pp. 265-279
Author(s):  
A. Corsi ◽  
A. L. Granata ◽  
O. Hudlicka
Keyword(s):  
Blood Supply ◽  
Periodic Acid ◽  
Capillary Density ◽  
Muscle Performance ◽  
Similar Proportion ◽  
Periodic Acid Schiff ◽  
Rat Soleus Muscle ◽  
Improved Performance ◽  
Isometric Twitch ◽  
Schiff Reaction

Muscle performance and structure was studied in rat soleus muscle with limited blood supply in combination with chronic muscle stimulation. Blood supply to the lower leg was restricted by ligation of the common iliac artery, electrodes were implanted in the vicinity of the sciatic nerve and ankle flexors were denervated. Three days later, soleus and gastrocnemius muscles were stimulated at 4 Hz four times a day for a period of 20 min with 2 h intervals between stimulations; this procedure was continued for 4 days. Muscle performance, histochemistry and ultrastructure were studied on the eighth day after operation in these muscles and in ischaemic unstimulated muscles with denervated ankle flexors. Both were compared with control animals. Muscles with limited blood supply developed less isometric twitch tension than control muscles (peak twitch tension in ischaemic muscle was 60.3 +/− 4.8 g g-1 muscle, mean +/− S.E.M., compared to 79.7 +/− 6.9 g g-1 in control muscle; tensions after 5 min contraction were 54.5 +/− 5.5 g g-1 in ischaemic muscle compared to 70.6 +/− 6 g g-1 in controls). Stimulated muscles with limited blood supply had higher peak (85 +/− 16.6 g g-1) and final (87 +/− 12 g g-1) tensions, and also fatigued less than muscles with limited blood supply but no stimulation. Histochemical estimation of capillary density (by staining for alkaline phosphatase) and slow (SO) and fast (FOG) fibres (by myosin ATPase staining) revealed similar capillary to fibre ratios (2.5) and a similar proportion of FOG fibres (around 18%) in all muscles. The proportion of glycogen-depleted fibres (estimated from the periodic acid Schiff reaction, PAS) in muscles removed from animals 10 min after a 5 min period of isometric twitches was significantly lower in ischaemic muscles (45.1 +/− 1.9%) than in control (80.5 +/− 1.5%) or chronically stimulated ischaemic muscles (67.3 +/− 4.0%). Electron microscopy showed disorganised myofibrils with Z-line streaming in 7.48 +/− 3.04% of fibres in muscles with limited blood supply. Swollen and degenerated mitochondria, dilated sarcoplasmic reticulum and areas of disrupted sarcolemma were also observed. Stimulated ligated muscles showed a significantly lower proportion of fibres with disorganised filaments (0.65 +/− 0.32%) and other signs of damage were much less frequent. The reduced damage and improved performance of chronically stimulated slow muscle may be the result of improved microcirculation, preventing accumulation of lactate.

scholarly journals Activation and activity of glycosylated KLKs 3, 4 and 11

2018 ◽  
Vol 399 (9) ◽  
pp. 1009-1022 ◽  
Author(s):  
Shihui Guo ◽  
Peter Briza ◽  
Viktor Magdolen ◽  
Hans Brandstetter ◽  
Peter Goettig
Keyword(s):  
Physiological Function ◽  
Periodic Acid ◽  
Hek293 Cells ◽  
Secretory Expression ◽  
Detailed Characterization ◽  
Periodic Acid Schiff ◽  
Clinical Biomarkers ◽  
Schiff Reaction

Abstract Human kallikrein-related peptidases 3, 4, 11, and KLK2, the activator of KLK3/PSA, belong to the prostatic group of the KLKs, whose major physiological function is semen liquefaction during the fertilization process. Notably, these KLKs are upregulated in prostate cancer and are used as clinical biomarkers or have been proposed as therapeutic targets. However, this potential awaits a detailed characterization of these proteases. In order to study glycosylated prostatic KLKs resembling the natural proteases, we used Leishmania (LEXSY) and HEK293 cells for secretory expression. Both systems allowed the subsequent purification of soluble pro-KLK zymogens with correct propeptides and of the mature forms. Periodic acid-Schiff reaction, enzymatic deglycosylation assays, and mass spectrometry confirmed the glycosylation of these KLKs. Activation of glycosylated pro-KLKs 4 and 11 turned out to be most efficient by glycosylated KLK2 and KLK4, respectively. By comparing the glycosylated prostatic KLKs with their non-glycosylated counterparts from Escherichia coli, it was observed that the N-glycans stabilize the KLK proteases and change their activation profiles and their enzymatic activity to some extent. The functional role of glycosylation in prostate-specific KLKs could pave the way to a deeper understanding of their biology and to medical applications.

Some Observations upon the Cytology of the Pars Distalis of the Surgically-removed Human Pituitary

1956 ◽  
Vol s3-97 (39) ◽  
pp. 379-391
Author(s):  
C. L. FOSTER
Keyword(s):  
Periodic Acid ◽  
Cell Types ◽  
Post Mortem ◽  
Β Cells ◽  
Human Pituitary ◽  
Periodic Acid Schiff ◽  
Golgi Bodies ◽  
Lipid Inclusions ◽  
Staining Methods ◽  
Human Anterior Pituitary

Human anterior pituitary tissue that had been removed at operation and immediately fixed was examined by a number of cytological and histochemical methods and by phase contrast and electron microscopy, and compared with similar material obtained post mortem. The general histological picture of good post-mortem material (not more than 4 hours post mortem) compared well with the surgically-removed tissue. For the study of silver impregnations of the Golgi substance, however, material removed at operation was found to be greatly superior. Evidence was obtained showing that the intracellular lipid inclusions seen post mortem were not artifacts resulting from cytolytic changes. There appeared to be no relationship between these lipid bodies and the Golgi material as revealed by the Aoyama method. No unequivocal dimorphism of the Golgi bodies, correlated with α- and β-cells, such as has been reported to occur in certain other mammals, was observed. Phospholipid was present in the granules of a substantial proportion of the α-cells. It was found that most of the cells which had been designated as β-cells after the application of certain routine staining methods, and most of the Gram-positive cells, reacted positively to the Periodic acid Schiff test: these cells could therefore be regarded as true β- or mucoid cells. A method for the demonstration in frozen sections of the cell-types, together with the lipid inclusions, is described.

Low doses of ethanol have Ca2+ ionophore-like effects on apical membrane potential of in vitro Necturus antrum

1991 ◽  
Vol 261 (1) ◽  
pp. G92-G103
Author(s):  
M. J. Rutten ◽  
C. D. Moore
Keyword(s):  
Membrane Potential ◽  
Apical Membrane ◽  
Periodic Acid ◽  
Ethanol Exposure ◽  
Membrane Potentials ◽  
Antral Mucosa ◽  
Low Doses ◽  
Periodic Acid Schiff ◽  
Induced Changes

The effects of low doses of luminal ethanol on the amiloride-sensitive apical membrane potential of Necturus antral mucosa were studied using conventional microelectrode techniques. Luminal ethanol (0.250-4.0% vol/vol) caused a dose-dependent hyperpolarization of the apical membrane potential (Vmc), an increase in transepithelial resistance (Rt) and resistance ratio (Ra/Rb), and a decrease in transepithelial potential (Vms). Luminal amiloride (100 microM) to 4% ethanol-treated antra did not cause any additional hyperpolarization of Vmc. Compared with luminal 2% ethanol-Ringer, an equivalent osmotic mannitol solution depolarized Vmc and basolateral potential (Vcs), decreased Rt and Ra/Rb, and increased Vms. A single dose of 0.50% ethanol attenuated the effects of a second 2% ethanol exposure on Vmc. No change in periodic acid-Schiff (PAS)-positive mucous granule content could be found between control and 2% ethanol-treated antra. The Ca2+ ionophores A23187 or ionomycin (0.25-5.0 microM) dose dependently hyperpolarized the Vmc and Vcs, increased Rt and Ra/Rb, and decreased Vms. Luminal Ca(2+)-free Ringer had no effect on luminal 2.00% ethanol-induced changes in membrane potentials or resistances. Pretreatment with BAPTA blocked by approximately 70 and 55% the Vmc hyperpolarization of 2 and 4% ethanol, respectively. Pretreatment with ruthenium red (10-50 microM) also dose dependently reduced the 2% ethanol-induced changes in Vmc. The data indicate that 1) low doses of luminal ethanol and Ca2+ ionophores have similar effects on Necturus gastric antral membrane potentials and resistances, 2) ethanol-induced hyperpolarizations of the Vmc are partially mediated through an alteration in intracellular Ca2+, and 3) low doses of luminal ethanol do not cause the release of antral epithelial mucous granules at the time when significant changes are occurring in the Vmc.

scholarly journals THE HISTOCHEMICAL INTERPRETATION OF THE COMPLEX RESULTS OF METHYLATION UPON GASTROINTESTINAL TRACT MUCINS, WITH SPECIAL REFERENCE TO THE PERIODIC ACID-SCHIFF REACTIVITY

1974 ◽  
Vol 22 (10) ◽  
pp. 986-991 ◽  
Author(s):  
P. E. REID ◽  
C. F. A. CULLING ◽  
W. L. DUNN
Keyword(s):  
Sialic Acid ◽  
Gastrointestinal Tract ◽  
Special Reference ◽  
Periodic Acid ◽  
Separate Study ◽  
Sulfate Ester ◽  
Periodic Acid Schiff ◽  
Smith Degradation ◽  
Vicinal Diol ◽  
Schiff Reaction

The histochemical use of methylation has complex results; particularly in respect of the periodic acid-Schiff reaction, these are analyzed and discussed. Methods are described which allow the separate study of the following effects: (a) the removal of the KOH/periodic acid-Schiff effect; (b) removal of sialic acid from a potential vicinal diol; and (c) the removal of O-sulfate ester from a potential vicinal diol. The use of the Smith degradation technique, in addition to the above, also allows inferences to be drawn in respect of the structure of the mucins (glycoproteins) being investigated.

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