Eficiencia diagnóstica de la citología nasal en pacientes con alergia respiratoria / Diagnostic efficiency of nasal cytology in respiratory allergic patients

Se realizó un estudio de eosinófilos en secreción nasal y de eosinófilos, basófilos y mastocitos en membrana de mucosa nasal, en 57 pacientes. A todos se le diagnosticó alergia respiratoria por la historia clínica, número de eosinófilos en sangre periférica, IgE sérica total, pruebas cutáneas y eventualmente por la dieta de eliminación y provocación. Diez niños sanos sirvieron como controles. Las muestras del moco se obtuvieron por barrido del piso de la nariz y del meato inferior. El material fue extendido en porta-objetos, fijado y coloreado por el método de Hansel. Las muestras de mucosa nasal fueron obtenidas del cornete inferior por medio de una cureta plástica descatable (Rhino-probe), extendidas sobre un porta-objetos, fijados y coloreados con Wright-Giemsa. La eficiencia del diagnóstico de la citología de las secreciones nasales, fue similar al de la mucosa, excepto en pacientes menores de un año de edad. Fue mejor la correlación entre atopía y citología nasal cuando ésta fue positiva tanto en las secreciones como en la mucosa

Eficiencia diagnóstica de la citología nasal en pacientes con alergia respiratoria / Diagnostic efficiency of nasal cytology in respiratory allergic patients

Se realizó un estudio de eosinófilos en secreción nasal y de eosinófilos, basófilos y mastocitos en membrana de mucosa nasal, en 57 pacientes. A todos se le diagnosticó alergia respiratoria por la historia clínica, número de eosinófilos en sangre periférica, IgE sérica total, pruebas cutáneas y eventualmente por la dieta de eliminación y provocación. Diez niños sanos sirvieron como controles. Las muestras del moco se obtuvieron por barrido del piso de la nariz y del meato inferior. El material fue extendido en porta-objetos, fijado y coloreado por el método de Hansel. Las muestras de mucosa nasal fueron obtenidas del cornete inferior por medio de una cureta plástica descatable (Rhino-probe), extendidas sobre un porta-objetos, fijados y coloreados con Wright-Giemsa. La eficiencia del diagnóstico de la citología de las secreciones nasales, fue similar al de la mucosa, excepto en pacientes menores de un año de edad. Fue mejor la correlación entre atopía y citología nasal cuando ésta fue positiva tanto en las secreciones como en la mucosa

Structure of the cumulus oophorus at the time of fertilization.

Ultrastructural and morphometric techniques were employed to examine the ovulated cumulus oophorus of hamsters and rats. Observations on cumuli prepared in a variety of ways including different chemical fixation techniques and cryofixation freeze substitution were compared. It was concluded that the cumulus mucus is not arranged in lamellae or granules as has previously been suggested but is composed of molecules which form very fine filaments when properly fixed. Morphometric analysis of cumuli fixed either in situ or after being explanted into medium revealed that the distance between neighboring cumulus cells was greater with increasing distance from the oocyte. Morphometry revealed that, when placed into medium, the cumulus expands possibly due to hydration. Thus physiological experiments carried out on cumuli should be performed very shortly after cumuli are isolated. From their ultrastructure cumulus cells appear to be actively involved in protein synthesis and secretion as well as steroid production.

Hymenolepis diminuta: intestinal goblet cell response to infection in male C57 mice.

Intestinal goblet cell numbers in two regions of the small intestine (20-30% and 60-70% distance form the pylorus) of male, 6- to 8-week-old C57 mice have been monitored following a 5-cysticercoid infection of the rat tapeworm, Hymenolepis diminuta. Test and sham-infected control mice were autopsied 0, 4, 8, 10, 14, and 28 days postprimary infection (p-1 degree-i) and 2, 4, 5, 7, and 14 days postsecondary infection (p-2 degree-i), administered 28 days p-1 degree-i. Results show a statistically significant increase in the number of mucus-containing goblet cells in both regions of the intestine during primary and secondary infections. Peak goblet cell numbers occurred on Day 8 p-1 degree-i and Day 5 p-2 degree-i in the 20-30% region and on Day 10 p-1 degree-i and Day 5 p-2 degree-i in the 60-70% region. In both regions, cell numbers declined to control levels by Day 14 p-1 degree-i, but remained significantly above control values 14 days p-2 degree-i. The increase in cell numbers correlated with an increase in goblet cell theca size and observable amounts of luminal mucus. The same infection regime in mice treated with cortisone elicited no goblet cell response. Male Wistar rats given a 10-cysticercoid infection and autopsied on Day 0, Day 10, and 15 months p-i showed a statistically significant increase in mucus-containing goblet cells only in the 60-70% region of intestine 10 days p-i; however, the worm burden was not eliminated. The functional significance of these results is discussed in relation to host immunity and murine cestode rejection.

Morphometric definition and grading of gastric intestinal metaplasia.

Type I and type III intestinal metaplasia in gastric mucosa have been examined using morphometric methods. Tissue (volume per cent gland, lumen, epithelium, goblet cell vacuoles) and nuclear parameters (area, with related standard deviation, and form factors) were used as indicators of gland crowding, nuclear-cytoplasmic ratio, nuclear atypia, and pleomorphism. In type III intestinal metaplasia, there is significantly (i) greater nuclear pleomorphism, (ii) a higher nuclear-cytoplasmic ratio, and (iii) smaller and less numerous goblet cell vacuoles in both the upper and the lower parts of the crypts. These two parameters have significantly higher values in the lower than in the upper part of individual crypts. No cell population with large pleomorphic nuclei characterized type III metaplasia, though there was more variation in nuclear size.

Characterization of ocular mucus extracts by crossed immunoelectrophoretic techniques.

Crossed immunoelectrophoresis (CIE) and crossed immunoelectrofocusing (CIEF) were used to characterize proteins and mucosubstance in the saline extract of human ocular mucus pooled from the normal eyes of donors. CIE resolved more components with greater specificity than previous techniques. Up to 25 components were identified. Lactoferrin, protein G, tear prealbumin, and ocular mucosubstance were found to be ocular-specific. CIE also allowed for the study of protein associations of: 1) albumin-alpha 1-antitrypsin; 2) albumin-tear prealbumin; and 3) IgA-secretory component and lactoferrin-mucosubstance. CIEF revealed that most of the proteins were in the pI range of 4.6-7.4. Up to 19 components were identified. Protein associations revealed by CIE were not evident by CIEF. These results provide a basis for future comparative analyses of tear and mucus from normal and diseased eyes, essential for a better understanding of tear and mucus function.

The effect of bromhexine hydrochloride on the viscoelastic properties of mucus from the mini-pig.

Mucus was collected daily from open-ended pouches established surgically in three mini-pigs. After a five day control period bromhexine hydrochloride (BHCl) was administered to each pig at dose levels of 0.5, 1.0 and 2.0 mg.kg-1 twice daily for five days. Each study period was followed by a five day washout period, when mucus was collected but no drug given. The viscoelastic properties of each mucus sample were determined using creep compliance analysis. BHCl was shown to reduce the residual shear viscosity (p less than 0.05) and increase the instantaneous shear compliance at all dose levels (p less than 0.005), despite the large inherent intra- and inter-animal variation in the rheological properties of the daily samples. No change was found in the wet weight of the mucus samples throughout any of the study periods. This experimental model would appear to provide a valuable in vivo method of assessing the mucoregulatory potential of administered compounds.

Relationship between milk intake and mucus production in adult volunteers challenged with rhinovirus-2.

In the first of three studies investigating the widely held belief that "milk produces mucus," 60 volunteers were challenged with rhinovirus-2, and daily respiratory symptoms and milk and dairy product intake records were kept over a 10-day period. Nasal secretion weights were obtained by weighing tissues collected and sealed immediately after use. Information was obtained on 51 subjects, yielding 510 person-days of observation. Subjects consumed zero to 11 glasses of milk per day (mean, 2.7; SE, 0.08), and secretion weights ranged from zero to 30.4 g/day (mean, 1.1; SE, 0.1). In response to an initial questionnaire, 27.5% reported the practice of reducing intake of milk or dairy products with a cold or named milk or dairy products as bad for colds. Of the latter group, 80% stated the reason as "producing more mucus/phlegm." Milk and dairy product intake was not associated with an increase in upper or lower respiratory tract symptoms of congestion or nasal secretion weight. A trend was observed for cough, when present, to be loose with increasing milk and dairy product intake; however, this effect was not statistically significant at the 5% level. Those who believe "milk makes mucus" or reduce milk intake with colds reported significantly more cough and congestion symptoms, but they did not produce higher levels of nasal secretions. We conclude that no statistically significant overall association can be detected between milk and dairy product intake and symptoms of mucus production in healthy adults, either asymptomatic or symptomatic, with rhinovirus infection.

[The x-ray microanalysis of the mucosa of the rat small intestine]. / Rentgenovskii mikroanaliz slizistoi tonkoi kishki krysy.

A rat small intestine mucosa is shown to accumulate significant amount of potassium and chloride. There was found a correlation between the content of these chemical elements and glycoprotein compartmentalization in goblet cell secret, brush border of enterocytes and a mucus layer. In this connection a role of mucus glycoproteins in membrane digestion is discussed. For preparation of samples the cryotechniques of electron microscopy are used.

The adherent gastric mucus gel barrier in man and changes in peptic ulceration.

The mucus barrier is a layer of water-insoluble gel adherent to the gastroduodenal epithelium. In man most previous studies have focused on luminal mucus or histological assessment of presecreted, intracellular mucus--neither of which can be directly correlated with the protective capacity of the adherent mucus barrier. We here describe direct observation of adherent mucus thickness in man, and changes in peptic ulceration. Adherent mucus gel on human antral mucosa is a continuous homogeneous layer of variable thickness, in the range 50-450 microns (median 180 microns), comprising 67% polymeric mucin. In gastric ulcer patients, adherent antral mucus is significantly increased in thickness (median 240 microns), but is very heterogeneous and structurally a substantially weaker gel, comprising only 35% polymeric mucin. Adherent antral mucus from duodenal ulcer patients is homogeneous, significantly thinner (median 110 microns), and structurally a weaker gel, comprising 50% polymeric mucin. The adherent mucus layer from patients with gastric carcinoma resembled that from subjects with gastric ulcer in that it was very heterogeneous, of significantly increased thickness (median 240 microns) and structurally a very weak gel (23% polymeric mucin). These results are discussed in the context of gastroduodenal mucosal protection against acid and pepsin in the gastric juice.

Ionized calcium in human male and female reproductive fluids: relationships to sperm motility.

The levels of ionized calcium in seminal plasma were approximately 20% of the serum levels. In contrast, cervical mucus contained a level of ionized calcium similar to both serum and follicular fluid. Titration of seminal plasma and serum with increasing concentrations of calcium chloride indicated a 10-fold higher calcium-binding capacity for seminal plasma. In a random group of men under semen investigation, concentrations of ionized calcium and citrate in semen were inversely correlated (r = 0.732; P less than 0.001), an observation which was confirmed by studies of split ejaculates. These findings supported the contention that citrate is the major regulator of the levels of ionized calcium in seminal plasma and primarily responsible for maintaining the calcium gradient between the seminal plasma and cervical mucus. No significant relationship could be demonstrated between the levels of ionized calcium in the ejaculates and any of the motility characteristics of the spermatozoa in the same sample. Furthermore, the addition of increasing quantities of calcium chloride (0.16-20.00 mM) to washed spermatozoa had no major effects on their progressive motility. These data suggest that human spermatozoa are effective in maintaining an appropriate level of internal ionized calcium, necessary for normal motility, despite fluctuations in external calcium.

The origin of DNA associated with mucus glycoproteins in cystic fibrosis sputum.

The relative importance of host and bacteria-derived deoxyribonucleic acid (DNA) in the increased viscoelasticity of purulent sputum in cystic fibrosis (CF) and other airway diseases is unclear. We report the identification of the DNA associated with mucus glycoproteins purified from the purulent sputum of 9 patients with CF. Mucus glycoproteins were purified from CF sputum by gel exclusion chromatography and the co-purifying DNA isolated by phenol extraction. Electrophoresis indicated that the DNA preparations had a size of approximately 300 to greater than 50,000 bases. The origin of the DNA was determined by slot blotting and subsequent hybridization with 32P-labelled DNA probes specific for human DNA sequences and those from bacterial species commonly isolated from CF sputum. The results indicated that in all cases the DNA was almost entirely human in origin. This implies that it is the patient's own DNA which may contribute to the rheological abnormalities of CF sputum.

Control of pH of airway surface liquid of the ferret trachea in vitro.

We measured the pH of airway surface liquid (ASL) secreted by the ferret trachea in vitro by using a catheter-tipped pH electrode implanted in a collecting cannula close to the airway epithelium. Mucus secretion was promoted by methacholine (0.02 mmol/l) in the organ bath. The pH of the ASL was 6.85 +/- 0.03 (SE) compared with a bath value of 7.39 +/- 0.01, when the bath was bubbled with 5.65% CO2. Changing the bath CO2 from 0 to 20.93% CO2 altered the bath pH from 8.06 to 6.96, but the ASL pH only varied from 6.92 to 6.85. This homeostasis of ASL pH was not the result of the buffering powers of the ASL, because ex situ buffer curves for secreted ASL were similar to those for Krebs-Henseleit solution. Changing the luminal CO2 content by blowing gases through the trachea changed ASL pH by values similar to that ex situ. However, when external organ bath CO2 was changed, the luminal CO2 changes were proportionately far smaller. Measurement of rates of diffusion of CO2 across the tracheal wall indicated that this was not a limiting factor in the results. Similarly, measurement of metabolic rate CO2 production in the tracheal lumen indicated that this did not significantly affect the results. We conclude that the pH of ASL is significantly on the acid side of the pH or interstitial fluid and plasma and that it is maintained relatively constant despite large changes in external pH.

Brucella abortus-specific immunoglobulin isotypes in serum and vaginal mucus from heifers vaccinated with Brucella abortus salt-extractable proteins and challenge exposed with virulent Brucella organisms.

Serum and vaginal Brucella-specific immunoglobulin isotypes (IgG1, IgG2, IgM, and IgA), obtained from 62 crossbred beef heifers vaccinated with Brucella abortus salt-extractable proteins and subsequently challenge exposed with B abortus S2308, were studied. Brucella-specific IgG antibodies and Brucella-specific immunoglobulin isotypes were quantitated by a fluorometric immunoassay. Serum and vaginal immunoglobulin responses were evaluated as a method of distinguishing infected from noninfected heifers. Rivanol precipitation, complement-fixation, buffered-antigen brucellosis tests and an ELISA were performed on sera. For immunoglobulin isotypes, vaccinated heifers had mean antibody responses higher than baseline mean antibody responses for at least 31 weeks after vaccination. After challenge exposure, significant differences (P greater than 0.05) were not detected between mean antibody responses of vaccinated and nonvaccinated heifers. Vaginal Brucella-specific antibody responses did not correlate with protection from disease. Vaginal Brucella-specific IgM was detected only at the time of abortion. Vaginal IgA appeared specific for identification of virulent B abortus infection. All serotests appeared adequate in distinguishing baseline titers from titers of heifers that had aborted and were considered bacteriologic culture-positive. Results of serotests neither consistently distinguished vaccinates from challenge-exposed cattle nor distinguished heifers that were challenge exposed, had aborted, and were considered bacteriologic culture-positive adequately from heifers that were challenge-exposed, had not aborted, and were considered bacteriologic culture-negative. Brucella-specific IgA appeared to be the most effective in distinguishing vaccinated heifers from challenge- exposed heifers and heifers that were challenge exposed and had aborted, from heifers that were challenge exposed and had not aborted. Brucella-specific serum IgA was detected up to 13 weeks after abortion.

Oral N-acetylcysteine or S-carboxymethylcysteine inhibit cigarette smoke-induced hypersecretion of mucus in rat larynx and trachea in situ.

Two weeks exposure of rats to cigarette smoke (CS) significantly (p less than 0.05) increased the secretion of fucose-containing glycoconjugates above normal in an in situ preparation of larynx and trachea. After equilibration mean basal secretion in CS-exposed rats was 24 micrograms (per 30 min collection) which was 8 times higher than that of unexposed animals (p less than 0.01). N-acetylcysteine (NAC) or S-carboxymethylcysteine (SCMC) given as 1% of the drinking water, before and after daily exposure to CS, significantly inhibited the development of the CS-induced increase in fucose secretion reducing the mean for basal secretion in each group to 7 and 5 micrograms, respectively (p less than 0.05). Neither NAC nor SCMC had significant effects on baseline glycoconjugate secretion in control animals. Albumin was inconsistently present in the secretions of both control and CS-exposed animals, whereas in those exposed to CS and also given one of the two cysteine derivatives there was a consistent increase in albumin transudation.

Morphology of the prairie dog gallbladder: normal characteristics and changes during early lithogenesis.

Studies were undertaken to describe the normal structure of the prairie dog gallbladder and adjacent cystic duct, and then to determine sequential changes that occurred as abnormalities in bile composition developed during high cholesterol feeding. Control animals were fed a diet with trace cholesterol, while experimental animals were fed a diet enriched with 1.2% cholesterol for 1, 2, 3, or 4 weeks. Light microscopy and scanning and transmission electron microscopy were used to characterize morphologic changes at each time interval. Biliary lipid composition was altered in all experimental groups, evidenced by significant decreases in bile-acid-to-cholesterol ratios. Cholesterol crystals appeared in experimental bile at 1 and 2 weeks, while stones formed at 3 and 4 weeks. The cystic duct and neck of the gallbladder occasionally displayed goblet cells. Little mucus was demonstrable in principal cells of the gallbladder, but much more in those lining the cystic duct. After 2 weeks of lithogenic diet, there was an increase in mucus content and secretion from all areas, as well as an influx of polymorphonuclear and mononuclear leukocytes. Accumulation of plasma cells in the lamina propria was an especially prominent feature of experimental tissues. These results suggest that 1) there is regional heterogeneity in the mucus content of the gallbladder and cystic duct of the prairie dog, and 2) both regions respond to lithogenesis with mucus hypersecretion and acute and chronic inflammatory changes prior to the appearance of cholesterol gallstones.

Evidence for the tight binding of human mucus proteinase inhibitor to highly glycosylated macromolecules in sputum.

Two extraction procedures of non-purulent sputum for the isolation of human mucus proteinase inhibitor (MPI) in its free and bound forms have been assayed. The dissociating procedure involved sputum homogenization in 1M NaCl and 4% (w/v) trichloroacetic treatment. When the soluble material was applied to a CM-Trisacryl column, a non-negligible, MPI-related inhibitory activity was recovered with the highly glycosylated constituents not retained on the column; the amount of MPI released in a free form was retained and eluted from the column according to the basic character of this inhibitor. The non-dissociating procedure consisted in a high water dilution (1:12) of sputum, known to bring into solution the macromolecular, fibrillar constituents, which was followed by ultrafiltration on selected Mr cut-off membranes. All the inhibitory activity was recovered with the high Mr (greater than 100,000) fraction which was shown on SDS-PAGE to be essentially composed of strongly glycosylated material; on electrophoretic analysis under non-reducing conditions, the MPI activity was visualized as three bands which corresponded to the inhibitor released from this high Mr fraction in the presence of SDS. As mucin-type molecules are the major, highly glycosylated constituents of bronchial secretions, it is suggested that they are responsible for the entrapping of MPI within their macromolecular network; it would appear that, as well as for lysozyme, electrostatic interactions occur between the acid charges of mucins and the basic charges of MPI. The possible in vivo consequences of these interactions on MPI activity are discussed.