[Characteristics of Lactobacillus strains contained in pharmaceuticals]. / Charakterystyka szczepów Lactobacillus wchodzacych w sklad preparatów farmaceutycznych.

The aim of the study was to characterize lactic acid bacteria (LAB) which are components of drugs administered orally in cases of intestinal disturbances, or antibiotic--related diarrhea. Biochemical properties, growth behavior, bile tolerance, and survival at low pH of six LAB strains (four strains L. rhamnosus and two L. acidophilus) were studied. The survival at low pH was determined in MRS broth (Difco) acidified to pH 1; 2; 3; and 4. Bile tolerance was tested on MRS broth with 0.3% oxgall (Difco). Between tested strains differences in ability to grow at low pH and survival in bile were observed. Only 0.01% inoculum of all examined strains survived at pH 1. Differences between strains in survival at low pH (pH 2 and pH 3) and tolerance of bile were observed. However, after 2 h incubation at pH 4, 100% of strains stayed alive. Examined strains demonstrated good 3% bile tolerance. All strains met the criteria for probiotic strains: ability to survive at pH 3 and in the presence of bile.

[Distribution of IgG subclasses and their biological activity in bioglobulin]. / Sklad podklas IgG i ich aktywnosc biologiczna w bioglobulinie.

Preparations of human immunoglobulin for intravenous use (IVIG) should contain proper percentage of IgG subclasses, corresponding to physiological proportions with preserved activity of antibodies. The amounts of IgG subclasses were determined using the radial immunodiffusion technique against to the WHO reference serum 67/97. It appears that variations are observed among the different lots and manufacturers of the various formulations of IVIG available in Poland. Although some of these differences are not statistically significant, significant differences in subclass concentrations were found between formulation of IVIG, with Bioglobulina being lower in IgG1 and higher in IgG2 and Endobulin having a lower per cent of IgG3. The study was aimed at establishment of activity values against diphtheria and tetanus toxoids and Streptococcus group B (GBS). IgG antibodies to tetanus and diphtheria toxoid antigens were predominantly of the IgG1 and IgG2 isotype. IgG4 subclass, was present in smaller proportion and IgG3 fractions was absent. Antibodies to GBS were only IgG2 and IgG1 isotypes. Basing on obtained results it can be stated that in spite of the imbalance between the amounts of IgG1 and IgG2 in Bioglobulina, the activity which was determined in subclasses is comparable with that of other IVIGs.

[Evaluation of antibacterial and antitoxic activity of normal human immunoglobulin for intravenous use (IVIG)]. / Ocena antybakteryjnej i antytoksycznej aktywnosci ludzkiej normalnej immunoglobuliny do stosowania dozylnego (IVIG).

In the present study, human immunoglobulins for intravenous use (IVIG), were tested for contents of diphtheria antibodies, antistreptolysin O, antistaphylolysin and antibody level of endotoxin of B. pertussis, enterobacterial common antigen and group B Streptococci. It was shown that all examined immunoglobulin preparation contained antibodies against all tested antigens. Our investigation revealed differences between IVIG preparations and IVIG lots. Basing on these results, we know that the biological activity of Bioglobulin is the same as biological activity of other IVIG preparations produced by foreign manufactures.

[Synergistic action of penicillin and immunoglobulin for intravenous (IVIG) use]. / Synergistyczne dzialanie penicyliny i immunoglobulin do stosowania dozylnego (IVIG).

Combined effects of intravenous immunoglobulin and antibiotic in killing bacteria are of interest with extending clinical use of IGIV. Since the agents differ in antibacterial activity exerted by each of them, and their influence on the other, it is difficult to evaluate the combined effect in vitro. In our experiments the titer of antibody, and the killing of B Streptococcus type 090R, phagocytosis by chemiluminescence were examined in opsonic mixture. This mixture consist of IVIG, fresh serum, and neutrophils plus penicillin or without. The effect of combined IVIG and penicillin was higher than the effects of separate activities on bacteria. This observations suggest that the combination of IVIG and penicillin potentially useful in the treatment of GBS infection in some cases.

Glucocorticoid regulation of an insulin-like growth factor-binding protein-4 protease produced by a rat neuronal cell line.

Insulin-like growth factor-binding protein-4 (IGFBP-4) is expressed in distinct regions in the rodent brain from the perinatal period into adulthood and is postulated to modulate the action of the insulin-like growth factors (IGFs) in vivo. This study was initiated to examine the regulation of IGF-binding protein-4 (IGFBP-4) in B104 cells, a rat neuronal cell line in which IGFBP-4 is the predominant secreted IGFBP. Exposure of B104 monolayer cultures to dexamethasone reduced native IGFBP-4 abundance to less than 10% of that in control medium by 48 h. Immunoblots showed that the decline in intact 24-kilodalton IGFBP-4 was accompanied by an increase in a 16-kilodalton immunoreactive fragment. In addition, IGFBP-4 proteolytic activity in medium was increased after exposure of the cells to dexamethasone. The protease was calcium dependent and appeared to be of the serine protease class, because activity could be inhibited by phenylmethylsulfonylfluoride and aprotinin, but not antipain, leupeptin, or pepstatin. Although the proteolytically modified IGFBP-4 retained the ability to bind IGFs, the affinities were approximately 13- and 20-fold lower for IGF-I and IGF-II, respectively. These data indicate that B104 cells produce an IGFBP-4 protease that is regulated by glucocorticoids. The actions of this protease reduce the affinity of IGFBP-4 for the IGFs without abolishing binding. Because both the IGFs and glucocorticoids have important roles in brain development, it is possible that some glucocorticoid actions in the brain could be mediated by proteolysis of IGFBP-4, which, in turn, would alter IGF action.

[Opsonizing activity of human immunoglobulin preparations for intravenous use (IVIG)]. / Aktywnosc opsonizujaca preparatów ludzkich immunoglobulin do stosowania dozylnego (IVIG).

It was shown that all the examined immunoglobulin preparations (IVIG) enhance growth of opsonization of S. aureus, S. epidermidis, E. coli which was studied using the chemiluminescence method. However, no opsonization growth was observed of P. aeruginosa rods with the participation of IVIG (except for Sandoglobulin and the specific immunoglobulin Pseudomonas). We assume that such a result was due to the P. aeruginosa strain's susceptibility to lysis factors of the human serum used in the experiment as the complement source. It was confirmed that phagocytosis with the participation of IVIG measured by the chemiluminescence test requires the presence of a complement. The bactericidal test showed that all IVIG preparations, Pentaglobin in particular, are bactericidal and active against P. aeruginosa. Basing on the results obtained, we believe that the biological activity of the Polish preparation is comparable with the investigated IVIG preparations from foreign firms.

[Content of IgE in preparations of human immunoglobulin for intravenous use (IVIG)]. / Zawartosc IgE w preparatach ludzkich immunoglobulin do stosowania dozylnego (IVIG).

The study was aimed at determination of concentration of total IgE and IgE specific for D. pteronyssinus in preparation IVIG Bioglobulin. Twelve series of Bioglobulin and 20 series (8 preparations) registered in Poland, were tested. It was found that all series of Bioglobulin contained IgE in amount from 34.5 iu/ml to 105 iu/ml. In only one series IgE specific against D. pteronyssinus was detected in amount of 0.93 j/ml. It is concluded that concentration of IgE in preparation Bioglobulin is comparable to amount of IgE in other IVIG preparations.

[Evaluation of contents of A and B group substances in biological preparations]. / Ocena zawartosci substancji grupowych krwi A i B w preparatach biologicznych.

The study was aimed at determination of blood A and B group substances in biological preparations used in Poland. Twenty three series were investigated, namely: Di-Te-Per, Ty-Te, Ty, Te, against cholera, vaccine according to Delbet and Panodin. Also were tested: 65 series of imported preparations of immunoglobulin g (i.v.) such as Endobulin, Sandoglobulin, Gamma-Venin, Veinoglobuline and 5 local series such as Bioglobulin, as well as 9 series of preparation LNI (i.m.) Human Gamma Globulin. Presence of substance A was detected in tetanus and botulinum horse antitoxins in amount from 3.75 micrograms/ml to 30 micrograms/ml. Group substances A and B contained 6 series of LNI preparations-Veinoglobuline. Amount of substance A was detected as 3.75 micrograms/ml-7.5 micrograms/ml and of substance B as 2,5 micrograms/ml-5 micrograms/ml. Group substances A and B were not present in vaccines used according to vaccination calendar.

[Heat stability of potency of bacterial vaccines and the titer of tetanus antitoxin of Polish production]. / Stabilnosc cieplna mocy szczepionek bakteryjnych oraz miano antytoksyny tezcowej produkcji krajowej.

Described the stability of potency vaccines (DTP, BCG) and immunoglobulins (human's and animal's) at storage and experimental temperatures. Thermal degradation rate and design of loss of potency in time have been determined by Arrhenius equation. Our results were similar to WHO data from preparations which have been made in another countries.

Characteristics of hepatic receptors for somatomedin-C/insulin-like growth factor I and insulin in the developing human.

Insulin and the somatomedins (Sms) are putative regulators of cell proliferation and metabolism in the fetus. Since the liver is a potential target tissue of these hormones during fetal life, we characterized the hepatic receptors for Sm-C/insulin like growth factor I (Sm-C/IGF-I) and insulin during the second trimester of human fetal development. Membrane-enriched fetal liver homogenates specifically bound 8.9 +/- 1.5% (+/- SD) of added [125I]insulin and 5.1-7.2% of [125I]Sm-C/IGF-I. Binding of both hormones was constant from 12-20 weeks gestation and was much greater than that in adult liver membranes. Analysis of dose-response data indicated high affinity between each receptor and its respective ligand (Kd for the Sm-C/IGF-I receptor, 2.2 X 10(-10) M; Kd for insulin receptor, 5.2 X 10(-10) and 7.7 X 10(-9) M). Limited cross-reactivity (approximately 1:1,000) of insulin with the Sm-C/IGF-I receptor and Sm-C/IGF-I with the insulin receptor was found. Affinity labeling studies showed that each receptor possessed an approximately 135,000-dalton subunit which was a part of a larger disulfide-linked complex. Thus, the human fetal liver has specific receptors for Sm-C/IGF-I and insulin that are similar to those described for other tissues in terms of both hormone-binding characteristics and subunit structure, suggesting that these receptors mediate important cellular functions at this stage of fetal development.

Changes in glucagon do not play an essential role in the glucoregulatory responses to mild hyperinsulinemia in dogs.

To examine whether an increase in the glucagon concentration is essential for restoring hepatic glucose output following moderate decrements in blood glucose, we used isotope dilution techniques in trained conscious dogs (n = 5) to measure glucose production (Ra) and glucose utilization (Rd) during mild hyperinsulinemia (19 +/- 1 mU/l). In Study A, when insulin was infused to raise plasma insulin (IRI) from 13 +/- 2 to 19 +/- 1 mU/l, basal glucose (93 +/- 3 mg/dl) fell at a rate of 0.37 +/- 0.06 mg/dl/min over 30 min. Ra fell from 2.8 +/- 0.4 mg/kg/min by 0.5 +/- 0.1 mg/kg/min at 20 min (P less than 0.05), but recovered to baseline by 30 min; glucagon (IRG) fell transiently but returned to baseline by 45 min. In Study B, endogenous secretion of IRI and IRG was suppressed by infusion of somatostatin (0.2 microgram/kg/min), while peripheral concentrations were maintained constant by replacing glucagon (0.65 ng/kg/min) and insulin (0.225 mU/kg/min). Steady-state baseline plasma IRI, IRG, glucose and glucose turnover rates were similar to Study A; hyperinsulinemia was then induced as in Study A. Glucose fell by 0.78 +/- 0.19 mg/dl/min over 30 min and, as in Study A, Ra decreased transiently, but recovered to baseline by 30 min. The restoration of Ra occurred in study B despite constant IRG, and preceded later increments in cortisol and catecholamines at 60-90 min. Thus, in both studies A and B, Ra recovered to baseline without an increase in IRG and before the onset of significant hypoglycemia (glucose 83 +/- 1 and 70 +/- 1 mg/dl).(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of fetal insulin infusion on glucose kinetics in pregnant sheep: a compartmental analysis.

A three-compartment model, consisting of fetus (F), uteroplacenta, and mother (M) was applied to quantitate the effects of fetal hyperinsulinemia on glucose kinetics in pregnant sheep late in gestation. The approach combines the Fick principle with isotope dilution of differentially labeled glucose isotopes, infused simultaneously to F [U-14C]- and M [2-3H]glucose. In the basal state, rates of umbilical glucose uptake (8.37 +/- 0.98 mg/kg per min) and fetal glucose utilization (7.38 +/- 1.13) were equivalent (mean +/- SE; n = 12). When fetal insulin was increased from 13.7 +/- 2.2 to a plateau of approximately 100 microU/ml, arterial glucose decreased from 18.9 +/- 0.8 to a new steady state of approximately 13 mg/dl (P less than 0.001). Whereas umbilical glucose uptake increased at 90 min and remained elevated thereafter (P less than 0.01), fetal glucose utilization increased only transiently at 60 min by 1.9 +/- 0.8 mg/kg per min (26%; P less than 0.05) and then returned to base line. Insulin's persistent effect, however, was evident from the sustained doubling of the glucose clearance rate from 39.3 +/- 5.9 to 66.6 +/- 10.5 ml/kg per min (P less than 0.005). No endogenous fetal glucose production was evident throughout the experiments. Maternal glucose production and utilization remained unchanged, although there was a small decline in M glucose concentration and an increase in glucose transfer from M to the uteroplacenta and F, from 33.9 +/- 8.1 to 48.1 +/- 7.0 mg/min at 60 min (P less than 0.01 by paired analysis). We conclude that fetal hyperinsulinemia initially lowers glucose concentration by transiently increasing fetal glucose utilization.(ABSTRACT TRUNCATED AT 250 WORDS)

Modulation of hepatic protein kinase activity by indomethacin.

We previously demonstrated that treatment with indomethacin in vivo significantly blunted the glucagon-induced glycemic response in the rat. This prostaglandin synthetase (cyclo-oxygenase) inhibitor also accentuated the evanescent effect of glucagon on hepatic glucose output in the intact, anesthetized rat. In this report, we present evidence that impairment of glucagon action in the rat liver by indomethacin is mediated through its inhibitory effect on both cAMP-dependent and cAMP-independent hepatic protein kinase. Indomethacin treatment did not have a measurable effect on any of the other components of the glucagon transducer system. Furthermore, infusion with glucagon for two hours that maintained plasma glucagon values at high physiological levels significantly reduced hepatic cAMP-dependent protein kinase activity without altering its Km. Glucagon infusion also down-regulated its own hepatic receptors and glucagon-stimulated cAMP production; prostaglandin E1-stimulated cAMP production was not affected. We concluded that prostaglandins may play a role in the regulation of hepatic protein kinases involved in the glucagon-stimulated glycogenolytic response and that glucagon-induced down-regulation extends at least to the hepatic protein kinases. However, a direct effect of indomethacin or protein kinase and the adenylate cyclase complex cannot be ruled out.