Effect of various antibiotics on the circulation of proteins between the blood and lymph in macro-organisms

The effect of tetracycline, amphotericin B and kefzol on distribution of some proteins between the blood and lymph of the thoracic duct was studied on rabbits. Tetracycline was injected intramuscularly in the form of hydrochloride dissolved in 2% novocain in a dose of 25 mg/kg once or daily for 7 and 20 days. Kefzol (sodium cephazolin) was injected intramuscularly in a single dose of 100 mg/kg. Amphotericin B was injected intravenously in a dose of 1000 Units/kg once or for 5 days. The lymph samples were collected from the thoracic duct of rabbits treated with single doses of the antibiotics 1 and 24 hours after their injection. When the animals were treated with the antibiotics repeatedly the lymph samples were collected 24 hours after the last injection. The level of the total protein and the ratio of the protein fractions, i. e. albumins, alpha 1-, alpha 2-, beta- and gamma-globulins in the lymph and blood serum were determined. On the basis of these findings the protein coefficient (albumin/globulin) of the lymph and blood, the coefficients of the protein permeability of the blood vessels (R) and the constants of selective permeability of the blood capillaries (S) were calculated. It was shown that the shifts in the protein circulation between the blood and lymph had mainly the same trends independent of the antibiotics used and their retention time in the host. A significant decrease in the permeability of the blood vessel walls in respect to the total protein and gamma-globulins and a marked increase in their selectivity in passing of the protein molecules of different size were observed in all cases.     

Circadian variations of the plasma proteins in the adult male rat under natural synchronisation (author's transl)]

Circadian rhythms of blood total proteins, albumin, alpha 1-, alpha 2-, beta- and gamma-globulins were documented in 80 Wistar SPF adult male rats, synchronized by natural light (06.00-18.00 h) and darkness (18.00-06.00 h) during the month of October 78. Blood was sampled at four fixed time points in the 24 h scale (i.e. 04.00, 10.00, 16.00 or 24.00 h). Total proteins, albumin, alpha 2- and gamma-globulins showed a statistically significant rhythm with a maximum at 04.00 h. The data obtained in anaesthetized rats under artificial synchronization and in man are compared, taking into account that the rat is a nocturnally active rodent. The present work partially confirms the hypotheses of previous chronopharmacological studies, thus e.g. curarizing substances have a mimimum activity when the protein plasma level, especially albumin, is the highest.     

Alternative changes of activity of alpha2-macroglobulin and alpha1-antitrypsin in rat blood following damage in capsaicin-sensitive nerves

Effects of functional ablation of peptidergic sensory nerves with neurotoxic doses of capsaicin (150 mg/kg, s/c) as well as of their stimulation with small doses of capsaicin (5 mg/kg, i/p) on activity of proteinase inhibitors: alpha1-antitrypsin (alpha1-AT)-serine proteinase inhibitor and alpha2-macroglobulin (alpha2-MG)-nonspecific inhibitor were investigated in rat blood. The present results indicate alternative changes in activity of these proteinase inhibitors after damage of capsaicin-sensitive nerves: increasing decline in activity of alpha1-AT 1 and 3 or 14 days after administration of capsaicin and increase in activity of alpha2-MG land 3 day after the injection. The stimulation of afferent nerves with capsaicin did not change activity of the proteinase inhibitors 1 and 24 hours after the injection. It is suggested that the stable decrease in activity of alpha1-AT during a long period after the damage of capsaicin-sensitive nerves indicates an important role for these nerves in the regulating alpha1-AT that may exert a tonic effect on the activity alpha1-AT.     

Patterns of changes in proteins in the peritoneal fluid of women during the periovulatory phase of the menstrual cycle

During a laparoscopy that was performed between Day -6 and Day +9 of the cycle as related to the day of the LH peak (Day 0), the peritoneal fluid of 100 healthy female volunteers of proven fertility was collected and analysed. Peritoneal fluid volume and concentrations of total protein, albumin, alpha 1-, alpha 2-, beta- and gamma-globulins, IgA, IgG, IgM, haptoglobulin, acid-alpha 1-glycoprotein, alpha 1-antitrypsin, alpha 2-macroglobulin, C3-, C4- and C-reactive protein were determined. The peritoneal fluid volume and the concentrations of most proteins analysed showed an increase during the post-ovulatory phase of the period investigated. The peritoneal fluid:serum ratio of each individual protein showed a significant inverse correlation with its molecular weight. This confirms the assumption that peritoneal fluid is mainly an exudation product, most probably of ovarian origin.     

Carbohydrate, protein and glycoprotein metabolism by the guinea-pig liver in chronic metribuzin poisoning

Male guinea-pigs weighing 400-600 g, 8 months old, were given metribuzin directly into the gastric lumen over a period of 30 days (20 animals) or 90 days (20 animals) 6 times a week. In the liver of the poisoned animals, the glycogen level and the AspAT and AlAT activities, while in the serum the total protein and the fractions albumin, alpha 1-globulin and gamma-globulin significantly decreased; serum glucose and the serum fractions alpha 2-globulin and beta-globulin, each showed an increase. The glycogen level in the liver, total protein, glucose as well as the alpha 1 and alpha 2 globulin fractions in the serum showed not appreciable difference between 30 and 90 days of intoxication. After 90 days of metribuzin treatment AspAT and AlAT dropped in the liver and rose in the serum, in comparison to the 30-day values. As to the parameters of glycoprotein metabolism, the intoxicated animals showed a significant decrease and increase in concentration of hexosamines and sialic acids in the liver and serum, respectively. Metribuzin intoxication also cause a significant decrease in activity of glucosamine phosphate isomerase and significant increase in activity of glycosidases in the liver. The results suggest that metribuzin disturbs the metabolism of carbohydrates, proteins and glycoproteins in the guinea-pig liver.     

Alpha 1-antitrypsin and serum albumin mRNA accumulation in normal, acute phase and ZZ human liver

Alpha 1-Antitrypsin and albumin mRNA levels of 4 human livers were assessed using a newly sequenced cDNA clone of the carboxyterminal third of alpha 1-antitrypsin and a previously cloned albumin cDNA sequence. The relative concentration of alpha 1-antitrypsin mRNA was the same in poly(A)-containing RNA isolated from acute phase (MM) and alpha1-antitrypsin deficient (ZZ) individuals. In the acute phase liver relative to the normal (MM) liver, total RNA extracts showed a marked decrease in albumin mRNA concentration but no increase in alpha 1-antitrypsin mRNA. The ZZ liver showed decreased total and poly(A)-containing RNA content but the same proportion of alpha 1-antitrypsin to albumin mRNA as in the normal (MM) liver. This supports other evidence that ZZ alpha 1-antitrypsin deficiency is due to a defect in polypeptide processing (secretion) rather than a deficiency in mRNA accumulation.     

Capsaicin-induced release of tachykinins: effects of enzyme inhibitors.

We studied the effects of neutral endopeptidase (NEP) and angiotensin-converting enzyme (ACE) inhibition on the airway responses and the recovery of endogenously released substance P- and neurokinin A-like immunoreactivities (SP-LI and NKA-LI) after tracheal injection of capsaicin in isolated guinea pig lungs superfused through the trachea. Capsaicin in doses from 10(-10) to 10(-7) mol induced a dose-dependent increase in airway opening pressure and release of SP-LI and NKA-LI. Airway opening pressure changes and the recovery of SP-LI and NKA-LI were significantly greater in lungs superfused with the NEP inhibitor SCH 32615 than in control lungs. ACE inhibition with captopril did not increase the mechanical response or the recovery of SP-LI compared with lungs not receiving captopril. In lungs from guinea pigs pretreated with high doses of capsaicin 7-10 days before study, a regimen designed to deplete endogenous tachykinins, there was a significant decrease in the content and release of NKA-LI and SP-LI. There were no detectable airway effects of acute capsaicin infusion even after doses of 10(-5) mol. Because NEP is important in modulating the airway effects of endogenously released tachykinins after tracheal infusion of capsaicin, but ACE is not, it seems likely that tracheal administration of capsaicin releases tachykinins from epithelial rather than endothelial loci.     

Induction of acute phase proteins by dexamethasone in rat hepatocyte primary cultures

The effect of dexamethasone on the synthesis of acute phase proteins has been studied in primary cultures of rat hepatocytes. In the absence of dexamethasone no detectable amounts of alpha 2-macroglobulin were synthesized by hepatocytes cultured for 1 day. alpha 2-Macroglobulin synthesis was induced by dexamethasone concentrations of 10(-8) M or higher with a maximum at a concentration of 10(-7) M. alpha 1-Acid glycoprotein was synthesized in the absence of dexamethasone; however, its synthesis was also greatly stimulated by dexamethasone concentrations of 10(-8)-10(-6) M. Synthesis of alpha 1-proteinase inhibitor was stimulated only 1.4-fold at a dexamethasone concentration of 10(-7) M. The kinetics of induction of alpha 2-macroglobulin and alpha 1-acid glycoprotein were studied at a dexamethasone concentration of 10(-7) M. After an initial lag phase of 3 h the synthesis of both proteins showed a steady increase during 2 days. Synthesis of albumin remained unchanged under these experimental conditions. Unlike alpha 2-macroglobulin and alpha 1-acid glycoprotein tyrosine aminotransferase activity increased already during the first 3 h of induction by dexamethasone with a maximum at 12 h followed by a slight decrease.     

Contrasting effects of prostaglandins E2 and F2 alpha on sensitivity of the human cough reflex.

Prostaglandins have been shown to influence the sensitivity of the cough reflex. To investigate putative mechanisms of this, we examined the effects of inhaled prostaglandins E2 (PGE2) and F2 alpha (PGF2 alpha) on human cough responses elicited by two challenges, low chloride solution and capsaicin, which may activate different neural pathways. Baseline cough challenges were followed after 2 h by five breaths of PGE2, PGF2 alpha, or citric acid as a control. Cough challenges were repeated after 1 min. Potentiation of capsaicin responses occurred after PGE2 (median increase 2 coughs/min, range 0-7, P less than 0.01) and PGF2 alpha (median increase 8 coughs/min, range -3 to 27, P less than 0.01) compared with control. The effect of PGF2 alpha was greater (P less than 0.05) than that of PGE2. Potentiation of low chloride responses also occurred after PGF2 alpha (median increase 7 coughs/2 min, range -1 to 19, P less than 0.01), but effects of PGE2 were insignificant against this challenge (median change -1 coughs/2 min, range -4 to 13). These data suggest that PGE2 and PGF2 alpha have different effects on the sensitivity of the human cough reflex, which may be relevant during airway disease.     

Change in concentrations of luteinizing hormone subunit messenger ribonucleic acids in the estrous cycle of beef cattle.

Changes in the concentrations of LH subunit messenger ribonucleic acids (mRNAs) and in the LH content of the anterior pituitary of beef cattle were studied during the estrous cycle. Japanese beef cows were classified according to the expected day of the estrous cycle: stage I (early-luteal phase, days 1-4; day 1=day of ovulation), stage II (early-mid-luteal phase, days 5-10), stage III (late-mid-luteal phase, days 11-17) and stage IV (follicular phase, days 18-20), according to the morphology of the ovaries. The anterior pituitaries of the cows were collected and the levels of alpha and LHbeta subunit mRNAs were determined by slot-blot analyses. The LH content of the anterior pituitary was measured by radioimmunoassay. The level of alpha subunit mRNA in the pituitary of cows was highest in stage I and decreased significantly by stage II (P<0.05); thereafter it tended to increase. The level of LHbeta subunit mRNA did not change significantly during the estrous cycle. The LH content of the pituitary of cows was low in stage I and tended to increase by stage II, then to decrease from stage II to III, and to increase significantly from stage III to IV (P<0.05). These results suggest that the highest levels of gene expressions of alpha subunit in the anterior pituitary occur in the early-luteal phase of beef cows, while the LH content is increased most in the follicular phase. The enhanced gene expressions of common alpha subunit in the early-luteal phase could be important in replenishing the bovine anterior pituitary with LH, which is depleted of hormone by the LH surge or the enhanced pulsatile release.     

Plasma proteins in young and adult Spermestes c. cucculatus (Swainson) (Aves - Ploceidae - Estrildinae)]

The blood plasma of Spermestes c. cucullatus (Swainson) analysed electrophoretically on agar shows seven fractions. Immunoelectrophoresis against an antispermestes rabbit serum reveals 16 fractions. Individual variation of the plasma protein fractions is very low, as is also the case for the mobility of the fractions. With age, the alpha-globulins disappear gradually. In the newly hatched bird, the protein variations are more pronounced than in adults. Most important are the lower relative content of albumin and a corresponding relatively higher content of beta- and gamma-globulins. There seems to exist a slight sexual dimorphism in albumin level, which is higher in males than in females. During the reproduction period, both sexes show a fall of the relative concentration of albumin. Injections of gonadotropine hormone produce an increase of the plasma-proteins level and a marked decrease of the relative concentration of albumin. The overall figure is comparable with that of birds in natural breeding condition.     

Possibilities of earlier evidence of the lead load

A total of 138 workers of a motorcar transportation plant were examined to show a degree of lead damage. The analysis of obtained results was conducted from the two points of view: 1) according to employment time in plant; 2) according to work groups. The following laboratory examinations were carried out: delta-aminolevulin acid and coproporphyrin urinary excretion; determination of serous transaminase of glutaminoxalacetic acid and glutamin and piruvic acid, leucinaminopeptidase, whole protein, albumin-globulin coefficient, protein fractions (albumin alpha 1-, alpha 2-, beta 1-, beta 2, gamma) as well as of dehydratase activity of delta-aminolevulin acid in erythrocytes. In conformity with literary data an increased urinary excretion of delta-aminolevulin acid and of coproporphyrin to a different degree, as well as a decrease of dehydratase content of delta-aminolevulin acid in erythrocytes were detected. In the authors' view attention is to be drawn to a minor or greater extent to proportional increase of leucinaminopeptidase content however, in all the age groups and all the professional categories. The results of present examinations are considered by authors an important and sensitive parameter to evidence of lead damage.     

Axonal effects of capsaicin: an electrophysiological study

The effects of axonally applied capsaicin on the discharge activity and compound action potential of the cat vagus, saphenous and phrenic nerves and the cervical sympathetic trunk were studied under in vivo and in vitro conditions. Application of capsaicin (10(-4) M) to the vagus, saphenous and phrenic nerves resulted in the appearance of intense discharge activity which reached its maximum after 3-4 min and lasted for 15-20 min. Parallel with the increase in discharge activity, elicited by orthodromic activation induced by capsaicin, the amplitudes of the A delta and C components of an antidromically evoked compound action potential were significantly reduced. After the excitatory action of capsaicin vanished, an increase in the latency and duration and a decrease in the amplitudes of the components of the compound action potential were observed which might have led to the development of a local block of impulse propagation. These changes proved to be reversible after the removal of capsaicin from the nerve. Compound action potentials recorded from the saphenous or vagus nerves pretreated with capsaicin 3-5 days before the experiments failed to show any significant changes. It is concluded that upon direct axonal application capsaicin results in the excitation of both A delta and C fibres which is followed by a nonspecific but reversible blockade of impulse propagation. The possible significance of these transient effects of axonally applied capsaicin in term of the development of the highly specific functional impairment occurring a few days after perineural capsaicin treatment remains to be elucidated.     

Plasma extravasation through neuronal stimulation in human nasal mucosa in the setting of allergic rhinitis

Sanico, Alvin M., Satsuki Atsuta, David Proud, and AlkisTogias. Plasma extravasation through neuronal stimulation in humannasal mucosa in the setting of allergic rhinitis. J. Appl. Physiol. 84(2): 537-543, 1998.We havepreviously shown that capsaicin nasal challenge in subjects withallergic rhinitis produces a dose-dependent increase in the albumincontent of nasal lavage fluids. In the present set of studies, wedetermined whether this observation represents plasma extravasationthat is neuronally mediated. To evaluate whether glandular secretionscontribute to the albumin increase in nasal lavage fluids, volunteerswith allergic rhinitis were pretreated with atropine or placebo before capsaicin challenge. Atropine significantly reduced the volume ofreturned lavage fluids and their lysozyme content but increased theiralbumin and fibrinogen content. To assess the contribution of sensorynerve stimulation, subjects with allergic rhinitis were pretreated in asecond study with lidocaine or placebo before capsaicin challenge.Lidocaine significantly attenuated the capsaicin-induced increases inthe volume of nasal lavage fluids, as well as their lysozyme andalbumin content. To rule out the possibility of a direct effect oflidocaine on blood vessels rather than on nerves, healthy subjects werepretreated in a third study with lidocaine or placebo before bradykininnasal challenge. Lidocaine did not affect the bradykinin-inducedincrease in the albumin content of nasal fluids. We conclude that, inallergic rhinitis, high-dose capsaicin induces plasma extravasation inthe human nose and that this effect is neuronally mediated. Thisprovides more definitive evidence that neurogenic inflammation canoccur in vivo in the human upper airway.

     

Involvement of CGRP in the inhibitory effect of rutaecarpine on vasoconstriction induced by anaphylaxis in guinea pig

Previous investigations have indicated that calcitonin gene-related peptide (CGRP), a principal transmitter in capsaicin-sensitive sensory nerves, could alleviate cardiac anaphylaxis injury. Rutaecarpine relaxes vascular smooth by stimulation of CGRP release via activation of vanilloid receptor subtype 1 (VR1). In the present study, we examined the role of capsaicin-sensitive sensory nerves in anaphylactic vessels and the effect of rutaecarpine on antigen-challenged constriction in the guinea pig isolated thoracic aorta. The aortas were challenged with 0.01 mg/ml bovine serum albumin, and the tension of aorta rings was continuously monitored. The amount of CGRP released from thoracic aortas was determined in the absence or presence of rutaecarpine. Antigen challenge caused a vasoconstrictor response concomitantly with an increase in the release of CGRP from the isolated thoracic aorta, and the vasoconstrictor responses were potentiated by CGRP8-37 (10 microM) or capsaicin (1 microM). Pretreatment with diphenhydramine (1 microM) markedly decreased antigen-challenged vasoconstriction. Acute application of capsaicin (0.03 or 0.1 microM) significantly inhibited vasoconstrictor responses. Pretreatment with rutaecarpine (10 or 30 microM) significantly increased CGRP release concomitantly with decrease in antigen-challenged vasoconstriction, which was abolished by CGRP8-37 (10 microM) or capsazepine (10 microM). The present results suggest that an increase in the release of CGRP during vascular anaphylaxis may be a beneficial compensatory response, and that rutaecarpine inhibits antigen-challenged vasoconstriction, which is related to stimulation of endogenous CGRP release via activation of VR1.     

Induction and Proliferation of Callus Derived from Fruits of Different Varieties of Capsicum annuum

Callus was initiated on Murashige and Skoog (MS) medium containing different combinations of growth regulators or different concentrations of vitamins from pericarp of six varieties of Capsicum annuum differing in their capsaicin content. Callus derived from pericarp of low capsaicin containing varieties was snowy white, friable and showed excellent growth. Callus initiation was delayed (10-15 days) in Punjab Lal, which had very high fruit capsaicin content (7.0 mg g^?1 DW). It also showed relatively slow proliferation although callus obtained was white and friable. Several different media were tried to improve the initiation and the proliferation of the callus in this variety. Callus growth improved greatly by doubling the concentration of MS salts. Initiation time was reduced to 4-6 days by adding 10 mg l^?1 NAA and 0.5 mg l^?1 Kin in MS medium. Other combinations of growth regulators or increase in concentration of vitamins or activated charcoal (0.1%) resulted in poor callus growth and callus texture. Of all media tried, MS medium containing 2 mg l^?1 2,4 D and 0.5 mg l^?1 Kin was the best for callus initiation and proliferation.     

Airway hyperresponsiveness to bronchoconstrictor challenge after wood smoke exposure in guinea pigs.

Prior airway exposure to wood smoke induces an increase in airway responsiveness to subsequent smoke inhalation in guinea pigs (Life Sci. 63: 1513, 1998; 66: 971, 2000). To further characterize this airway hyperreactivity, we investigated and compared the airway responsiveness to bronchoconstrictor challenge before and 30 min after sham air exposure or wood smoke exposure in anesthetized and artificially ventilated guinea pigs. Various doses of substance P (0.8-6.4 microg/kg), capsaicin (0.2-3.2 microg/kg), prostaglandin F2alpha (30-3000 microg/kg), histamine (1-8 microg/kg), or acetylcholine (5-20 microg/kg) were intravenously injected at 2-min intervals in successively increasing doses to obtain the dose required to provoke a 200% increase in baseline total lung resistance (ED200). Wood smoke exposure significantly lowered the ED200 of substance P, capsaicin, and prostaglandin F2alpha whereas sham air exposure failed to do so. Furthermore, wood smoke exposure did not significantly alter the ED200 of histamine or acetylcholine. Pretreatment with phosphoramidon (2 mg/kg), an inhibitor of the neutral endopeptidase (the major degradation enzyme of substance P), before smoke exposure did not significantly affect the smoke-induced reduction in ED200 of substance P. Sectioning both cervical vagi before smoke exposure did not significantly alter the smoke-induced reduction in ED200 of capsaicin or prostaglandin F2alpha. These results suggest that airway exposure to wood smoke acutely produces airway hyperresponsiveness to substance P, capsaicin, and prostaglandin F2alpha, but not to histamine or acetylcholine. Since the combination of phosphoramidon and wood smoke exposure did not result in an additive potentiation of smoke-induced airway hyperresponsiveness to substance P, it is suggested that an inhibition of the degradation enzyme of substance P may contribute to this increase in airway reactivity. Furthermore, vagally-mediated bronchoconstriction does not play a vital role in enhanced airway responsiveness to capsaicin or prostaglandin F2alpha.     

Dynamics of physiological parameters in the nestling of black-backed gull Larus marinus experimentally infested by the cestode Microsomacanthus ductilus (Cestoda: Hymenolepididae)

The effect of the invasion with the cestode Microsomacanthus ductilus on physiological and biochemical processes in black-backed gull Larus marinus was examined. Experimental invasion of the gull nestling by the cestodes has been performed. Dynamics of the protein, lipid, and carbohydrate metabolism in the time history of the invasion was observed, in comparison with noninfested nestling. Increasing of the content of alpha-globulins and decreasing of the content of protein and albumin in the blood plasma of experimentally infested birds were registered to 4th day after invasion. To 7th day after invasion the level of general lipids and phospholipids decreases, while the content of gamma-globulins and modified form of albumin increases. To 10th day after invasion symptoms of intoxication were observed, but some parameters proved to be reverted to normal condition. So, it can be assumed, that the most intensive reorganization of the metabolism in infested birds takes place in the period between 4th and 7th days after infestation. Possible causes of the observed phenomena are discussed.     

Interaction of serum proteins with CYP isoforms in human liver microsomes: inhibitory effects of human and bovine albumin,alpha-globulins,alpha-1-acid glycoproteins and gamma-globulins on CYP2C19 and CYP2D6

The effects of serum proteins on the in vitro hydroxylation pathways of mephenytoin (CYP2C19) and debrisoquine (CYP2D6) were studied to enhance the predictability of in vivo drug metabolism from in vitro assays. Both CYP substrates are known to be weakly bound to albumin and the applicability of the free drug hypothesis was further appraised. Since bovine serum albumin (BSA) is used widely in in vitro assays, a comparison between human and bovine proteins was made. Four major serum proteins were studied: albumin, alpha1-acid glycoprotein (AGP), alpha- and gamma-globulins. Human serum albumin (HSA) inhibited both CYP activities about 20% more than BSA. The addition of human alpha-globulins, but not the bovine protein, resulted in marked reduction of 86% and 41% in CYP2C19 and CYP2D6 activities, respectively. This reduction of activity was strikingly greater than the fraction bound (14 and 22%, respectively). The inhibition was of the competitive type and the Ki values of human alpha-globulins on CYP2C19 and CYP2D6 were found to be 0.45% (4.5 mg/ml) and 3.5% (35 mg/ml), respectively. The effect of both human and bovine gamma-globulins on CYP isoforms was negligible. The Ki values of human and bovine AGP for CYP2C19 were 1.84% (420 microM) and 0.93% (210 microM), respectively. For HSA, human alpha-globulins and human and bovine AGP, the strongly decreased CYP activities in vitro cannot be explained by the free drug hypothesis. A direct interaction of these serum proteins with CYP enzymes is postulated. Differential effects of bovine and human serum proteins and CYP specific inhibition were observed.