A combination of the probiotic and prebiotic product can prevent the germination of Clostridium difficile spores and infection.

Clostridium difficile infection (CDI) is one of the most prevalent healthcare associated infections in hospitals and nursing homes. Different approaches are used for prevention of CDI. Absence of intestinal lactobacilli and bifidobacteria has been associated with C. difficile colonization in hospitalized patients. Our aim was to test a) the susceptibility of C. difficile strains of different origin and the intestinal probiotic Lactobacillus plantarum Inducia (DSM 21379) to various antimicrobial preparations incl. metronidazole, vancomycin; b) the susceptibility of C. difficile strains to antagonistic effects of the probiotic L. plantarum Inducia, prebiotic xylitol (Xyl) and their combination as a synbiotic (Syn) product; c) the suppression of germination of C. difficile spores in vitro and in vivo in animal model of C. difficile infection with Inducia, Xyl and Syn treatment. The VPI strain 10463 (ATCC 43255), epidemic strain (M 13042) and clinical isolates (n = 12) of C. difficile from Norway and Estonia were susceptible and contrarily L. plantarum Inducia resistant to vancomycin, metronidazole and ciprofloxacin. The intact cells of Inducia, natural and neutralized cell free supernatant inhibited in vitro the growth of tested C. difficile reference strain VPI and Estonian and Norwegian clinical isolates of C. difficile after co-cultivation. This effect against C. difficile sustained in liquid media under ampicillin (0.75 µg/ml) and Xyl (5%) application. Further, incubation of Inducia in the media with 5% Xyl fully stopped germination of spores of C. difficile VPI strain after 48 h. In infection model the 48 hamsters were administered ampicillin (30 mg/kg) and 10-30 spores of C. difficile VPI strain. They also received five days before and after the challenge a pretreatment with a synbiotic (single daily dose of L. plantarum Inducia 1 ml of 1010 CFU/ml and 20% xylitol in 1 ml by orogastric gavage). The survival rate of hamsters was increased to 78% compared to 13% (p = 0.003) survival rate of hamsters who received no treatment. When administered Xyl the survival rate of hamsters reached 56% vs.13% (p = 0.06). In both Syn (6/9, p = 0.003) and Xyl (3/9, p = 0.042) groups the number of animals not colonized with C. difficile significantly increased. In conclusion, the combination of xylitol with L. plantarum Inducia suppresses the germination of spores and outgrowth into vegetative toxin producing cells of C. difficile and reduces the colonization of gut with the pathogen. Putative therapeutical approach includes usage of the synbiotic during antimicrobial therapy for prevention of CDI and its potential to reduce recurrences of CDI.

The role of ADRB2 in myasthenia: genetic and immunological factors.

To examine the possibility of using ADRB2 adrenergic receptor agonists for correction of myasthenia-induced immunological and motor disturbances, a method of anti-ADRB2 antibody assay by ELISA was developed. The haplotype of ADRB2 gene was determined by MLPA-PCR in 34 myasthenia patients; blood sera from 40 patients were examined. The content of antibodies against ADRB2 receptors was elevated in 12 of 30 patients with generalized form of myasthenia. Arg-Arg in position 16 in ADRB2 receptors determines the developmental risk of side effects, while Gly-Gly in the same codon determines tolerance to ß2-adrenoceptor agonists and is associated with more severe course of myasthenia and resistance to ß2-adrenoceptor agonists treatment with ADRB2 agonists. The study demonstrated the possibility of using ADRB2 obtained gene engineering technique for quantitatively assay the autoantibodies.

Role of antibodies to neuronal α7-acetylcholine receptors in myasthenia.

The role of antibodies to a fragment of neuronal acetylcholine receptor was studied by EIA in patients with myasthenia. Antibody levels were significantly higher in patients with generalized myasthenia. Enzyme immunoassay of antibodies by the reaction with acetylcholine receptor fragment can serve as an additional method for studies of autoimmune myasthenia pathogenesis.

Role sympathetic autonomic nervous system in the regulation of immune response during myasthenia.

In 12 patients with myasthenia, the content of ß(2)-adrenoreceptors on the cell surface and activity of intracellular lymphocytic enzymes were determined by EIA and biochemical methods, respectively. In comparison with the normal, these patients demonstrated pronounced elevation in the content of ß(2)-adrenoreceptors and significant changes in activity of lymphocytic enzymes. In 10 of 12 patients, administration of the agonists to ß(2)-adrenoreceptors resulted in health improvement accompanied by normalization of EMG and immunobiochemical indices. Our findings suggest that intra- and intercellular signaling pathways and their modification can serve as potential targets for the therapy.

Impact of empiric antibiotic regimen on bowel colonization in neonates with suspected early onset sepsis.

The purpose of this study was to compare the impact of ampicillin and penicillin used for empiric treatment of early onset sepsis (EOS) on initial gut colonization by aerobic and facultative anaerobic microorganisms. A cluster-randomized, two-center, switch-over study was conducted in two paediatric intensive care units in Estonia and included 276 neonates. Rectal swabs were collected twice a week until discharge or day 60. Colonizing microbes were identified on species level and tested for ampicillin resistance (AR). The number of patients colonized with Gram negative microorganisms and Candida spp was similar in both treatment arms but ampicillin resulted in longer colonization duration (CD) of K. pneumonia (p = 0.012), AR Serratia spp (p = 0.012) and Candida spp (p = 0.02) and penicillin in that of AR Acinetobacter spp (p = 0.001). As for Gram positive microorganisms penicillin treatment was associated with a greater number of colonized patients and higher CD of Enterococcus spp and S. aureus but lower ones of S. haemolyticus and S. hominis. Influence of ampicillin and penicillin on initial gut colonization is somewhat different but these differences are of low clinical relevance and should not be a limiting step when choosing between these two antibiotics for the empiric treatment of EOS.

Surveillance of antimicrobial resistance of invasive pathogens: the Estonian experience.

The aim of the present study was to evaluate the needs for surveillance of invasive Gram-negative pathogens in Estonia. The antimicrobial susceptibility data of invasive isolates of Acinetobacter baumannii, Pseudomonas aeruginosa, Klebsiella spp, Escherichia coli, Staphylococcus aureus, Streptococcus pneumoniae and enterococci were collected in accordance with EARSS (European Antimicrobial Resistance Surveillance System) protocols. Despite the higher rate of Gram positive pathogens, their resistance to antimicrobials was low in contrast to the elevated resistance established for Gram negative pathogens. The higher resistance to antimicrobials was particularly associated with A. baumannii and P. aeruginosa. Also, the proportion of extended spectrum betalactamase (ESBL)-producing strains was 23% among Klebsiella spp. and 3.6% among E. coli. The inclusion of invasive Gram negative pathogens in antimicrobial resistance surveillance provides useful information concerning local pathogen susceptibility, as well as for the empirical treatment of suspected infections.

Surveillance of antimicrobial resistance of invasive pathogens: the Estonian experience.

The aim of the present study was to evaluate the needs for surveillance of invasive Gram-negative pathogens in Estonia. The antimicrobial susceptibility data of invasive isolates of Acinetobacter baumannii, Pseudomonas aeruginosa, Klebsiella spp, Escherichia coli, Staphylococcus aureus, Streptococcus pneumoniae and enterococci were collected in accordance with EARSS (European Antimicrobial Resistance Surveillance System) protocols. Despite the higher rate of Gram positive pathogens, their resistance to antimicrobials was low in contrast to the elevated resistance established for Gram negative pathogens. The higher resistance to antimicrobials was particularly associated with A. baumannii and P. aeruginosa. Also, the proportion of extended spectrum betalactamase (ESBL)-producing strains was 23% among Klebsiella spp. and 3.6% among E. coli. The inclusion of invasive Gram negative pathogens in antimicrobial resistance surveillance provides useful information concerning local pathogen susceptibility, as well as for the empirical treatment of suspected infections.

["Light" epithelial cells of swine and bovine oviducts]. / Issledovanie "svetlykh" kletok épiteliia matochnykh trub svin'i i korovy.

The ultrastructure of oviduct epithelium of clinically healthy cows and 15 sows was investigated using scanning and transmission electron microscopy. In all parts of the oviduct, ciliated and non-ciliated epithelial cells are present, but their number varies in both the investigated animals in different regions of the oviduct, depending on the phase of the estrous cycle. In addition to ciliated cells with numerous cilia on their luminal surface, so-called pale ciliary cells were found in all parts of the oviduct of cows and sows. The cytoplasm of these cells is electron-lucent, their luminal surface carries few cilia and short microvilli. The apical cytoplasm contains species specific secretory granules, which means that these cells have features characteristic of both secretory and ciliated cells. It is suggested that the pale ciliated and non-ciliated secretory cells are functional stages of the same tubar epithelium cell, and that the transformation between these two cell types is regulated by functional requirements of the organ in different phases of the estrous cycle.

[Long-term changes in the rat gastric mucosa after truncal vagotomy and pyloroplasty]. / Izmeneniia slizistoi obolochki zheludka u krys v otdalennye sroki posle stvolovoi vagotomii i piloroplastiki.

In 40 male mature albino Wistar rats histological changes of gastric mucosa were analyzed, parietal and chief cells were counted and gastric juice pH was determined 120, 240 and 365 days after truncal vagotomy and pyloroplasty. In all the groups gastric mucosa was attenuated and morphological signs of preatrophic gastritis (decline in parietal cells number, a certain mucoidization of fundal glands without changes in surface epithelium, alteration of mucosal lamina propria) were present. On the background of atrophic changes compensatory and adaptive processes (increase of mitotic activity of epitheliocytes--precursors of differentiating cells, normalization of parietal cells differentiation along the stem line, i.e. of ratio of differentiating, mature functioning and degenerating cell forms) were noted, that lead to repair of histo-structure and function on the whole, although at a lower level. Following long-term intervals after vagotomy glandular gastric epithelium acquires its histo-typical form with parietal secretory activity decreased to some extent.

[Electroneuromyographic study of the spastic-ataxic syndrome in multiple sclerosis]. / Elektroneiromiograficheskoe izuchenie spastiko-atakticheskogo sindroma pri rasseiannom skleroze.

Electroneuromyographical studies were performed in 39 patients with atactic-spastic and mixed forms of disseminated sclerosis. The examinations detected a drop in the velocity of impulse conduction along the tibial nerve in spastic forms and an increase in the velocity of impulse conduction along the medium nerve in atactic forms. The most pronounced were changes in velocity of afferent conduction. The data obtained were compared with electroneuromyographical studies in atactic and spastic syndromes of different origin.

[Syndrome of congenital muscular hypotonia in children ("flabby child" syndrome)]. / Sindrom vrozhdennoi myshechnoi gipotonii u detei (sindrom "vialyi rebenok").

Clinico-electroneuromyographic examinations of 108 children with the "flabby child" syndrome of various genesis were carried out. A classification of diseases accompanied with muscular hypotonicity in early childhood is suggested. The electroneuromyographic examinations revealed an increase of the speed of impulse transmission along the peripheral nerves, this transmission being the most pronounced in the first two years of the life, and levelled down by the age of 3 to 7 years.

[Short-latency brain stem evoked potentials to acoustic stimulation in closed craniocerebral trauma in children]. / Korotkolatentnye stvolovye vyzvannye potentsialy na akusticheskuiu stimuliatsiiu pri zakrytoi cherepno-mozgovoi travme u detei.

Twenty-six children with closed craniocerebral trauma (CCCT) of various severity were examined by recording short-latent stem-induced potentials in response to acoustic stimulation. Prognostical authenticity of the method in severe CCCT in children was established, which allowed reversible and irreversible disorders of brain them function to be distinguished. Studies in dynamics made it possible to follow the course of the pathological process and correct the applied therapy in time.

Mucosal surveillance cultures in predicting Gram-negative late-onset sepsis in neonatal intensive care units.

This study aimed to examine the spectrum and time course of gut and nasopharyngeal colonization with Gram-negative micro-organisms, and to define the value of surveillance cultures in predicting late-onset sepsis in neonates admitted to neonatal intensive care units. Nasopharyngeal and rectal swabs were collected on admission and twice weekly thereafter in 278 neonates admitted within the first 72 h of life with risk factors of early-onset sepsis. Sterile body fluid cultures were obtained on admission and subsequently as clinically indicated. Approximately half of the rectal (693/1250, 55%) and nasopharyngeal (558/1153, 48%) samples but only 6% (32/555) of the sterile fluid samples in 26 patients were culture positive for Gram-negative organisms. In total, 2108 invasive and mucosal culture pairs were analysed. The overall sensitivity, specificity, and positive and negative predictive values of a mucosal sample to predict late-onset sepsis were 27%, 66%, 4% and 94%, respectively. Patients with pre-existing colonization with Klebsiella pneumoniae (P = 0.011), Klebsiella oxytoca (P = 0.002), Escherichia coli (P = 0.003), Stenotrophomonas spp. (P = 0.003) and Pseudomonas spp. (P ≤ 0.001) were more likely to develop late-onset sepsis. No such association was found for Acinetobacter baumannii, Serratia spp. or Enterobacter cloacae. In conclusion, routine mucosal cultures are inefficient for the prediction of Gram-negative late-onset sepsis in neonatal intensive care units. However, targeted screening for specific organisms in an outbreak (e.g. Klebsiella spp., E. coli, Stenotrophomonas spp. and Pseudomonas spp.) may offer an opportunity to improve infection control measures and enable timely initiation of appropriate antibiotic therapy.

Growth during the first 6 months of life in infants using formula enriched with Lactobacillus rhamnosus GG: double-blind, randomized trial.

BACKGROUND: Probiotic bacteria have beneficial effects on the immune system and gastrointestinal tract, but the impacts of their long-term consumption on health and growth in early infancy are not well documented. The aim of this study was to evaluate the influence of Lactobacillus rhamnosus GG (LGG)-enriched formula on growth and faecal microflora during the first 6 months of life in normal healthy infants. MATERIALS AND METHODS: One hundred and twenty healthy infants (up to 2 months) received LGG-supplemented formula or regular formula in a double-blind, randomized manner until the age of 6 months. Weight, length and head circumference were measured monthly and transformed into standard deviation scores (SDS). Faecal samples were obtained from a random sample of infants (n=25) at entry and at the end of the study. RESULTS: One hundred and five infants (51 in the LGG group) completed the study. Children receiving LGG-supplemented formula grew better: their changes in their length and weight SDS (DeltaSDS) at the end of the study were significantly higher than those receiving regular formula (0.44+/- 0.37 versus 0.07+/- 0.06, P< 0.01 and 0.44+/- 0.19 versus 0.07+/- 0.06, P< 0.005, respectively). The LGG group had a significant, higher defecation frequency 9.1+/-2.06 versus 8.0+/- 2.8 (P<0.05). More frequent colonization with lactobacilli was found in the LGG group, 91% versus 76% (P<0.05) at the end of the study. CONCLUSIONS Infants fed with LGG-enriched formula grew better than those fed with regular formula. Further studies are necessary to clarify the mechanism of LGG in infant growth.

Differential diagnosis of various forms of myasthenia and endocrine ophthalmopathy by immunoblotting.

Immunoblotting can be used for screening a population of antibodies to acetylcholine receptor subunits circulating in the blot of patients with myasthenia. Torpedo Californica acetylcholine receptor served as the antigen. We found that in generalized myasthenia autoantibodies bind to alpha1- or alpha1- and gamma-subunits, while in ophthalmic form they bind only gamma-subunit of acetylcholine receptor. No antibodies to any of the acetylcholine receptor subunits were detected in patients with endocrine ophthalmopathy and in healthy volunteers. This method can be used for differential diagnosis of ophthalmic myasthenia and endocrine ophthalmopathy and for predicting generalization of the pathological process in patients with myasthenia.

Intestinal microbiota and immunoglobulin E responses in 5-year-old Estonian children.

BACKGROUND: Over the last few decades, several studies from different parts of the world have indicated an increasing prevalence of allergic diseases. This has been related to environmental factors, like changes of microbial pressure. Our previous studies have demonstrated differences in the intestinal microbiota between allergic and non-allergic children. AIM: To test the hypothesis that the intestinal microbiota and IgE response are related, both in allergic and non-allergic 5-year-old Estonian children. METHODS: The study group comprised 19 allergic and 19 non-allergic 5-year-old children, selected from a larger group who had been followed from birth. The diagnosis of allergy was based on clinical examination of the children and on data obtained from the questionnaires. The faecal microbiota were quantified by seeding serial dilutions on nine different media for incubation in different environment. The composition of the gut microbiota was expressed both as absolute counts of the various species and their relative share among the total counts of identified microbiota. RESULTS: Bifidobacteria were less commonly detected in children with allergic diseases than in healthy children and clostridia comprised a higher proportion among their gut microbes. Children with specific IgE antibodies to defined allergens had higher counts of clostridia and the counts of clostridia correlated with the level of serum IgE, but only so in allergic children. In non-allergic children, the serum IgE levels showed a positive correlation with the counts of bacteroides. CONCLUSION: The development of allergic diseases seems to be associated with the composition of the gut microbial ecosystem. High counts of potential pathogens, such as clostridia, are associated with clinical manifestations of allergy and IgE antibody formation.

Immunobiochemical characteristics of IgG antibodies in myasthenia.

The abzyme activity of mAb35 and pQ1-209 was compared with that of normal rabbit IgG and serum IgG from patients with various forms of myasthenia. It was found that mAb35 and pQ1-209 and IgG from patients with myasthenia possess catalytic activity. IgG from myasthenia patients with thymomas possess creatine phosphokinase activity, which 2-fold surpassed the control.

The effects of the nitric oxide synthase inhibitor 7-nitroindazole on the behaviour of mice after chronic ethanol administration.

The effects of the nitric oxide synthase (NOS) inhibitor 7-nitroindazole (7-NI) on the behaviour of mice after chronic and acute ethanol administration were studied. Male albino mice received ethanol by inhalation for 25 days. The plus-maze and staircase tests were carried out with control, ethanol-intoxicated and ethanol-withdrawn mice (7.5 h after the end of ethanol administration). The administration of NOS inhibitor 7-NI [20.0 mg/kg, intraperitoneally (i.p.)] 60 min or 7.5 h before the plus-maze test induced an anxiolytic effect in control mice. Chronic ethanol administration induced an anxiolytic, and ethanol withdrawal an anxiogenic, effect in mice. The administration of 7-NI (20.0 mg/kg, i.p.) caused behavioural depression in ethanol-intoxicated mice, but had no effect on the behaviour of ethanol-withdrawn mice. 7-NI had no effect on the behaviour of control mice in the staircase test. Chronic ethanol administration increased, and ethanol withdrawal decreased, the locomotor activity of mice in the staircase test. Likewise, in the plus-maze test, administration of 7-NI caused behavioural depression in ethanol-intoxicated mice, but had no effect on the behaviour of ethanol-withdrawn mice. In additional experiments, vehicle or 7-NI (20.0-120.0 mg/kg, i.p.) were administered 30 min before ethanol (3.0 g/kg, i.p.). 7-NI dose-dependently increased the duration of ethanol-induced sleep and inhibited ethanol clearance. On the basis of these data we can propose that the NO system has no major role in behavioural changes caused by ethanol withdrawal. At the same time NOS inhibitors can cause synergistic CNS depression with ethanol.

[Clinical significance of quantitative antibodies measurement in blood serum by interaction with muscle acetylcholine receptor domain in myasthenia patients]. / Klinicheskoe znachenie kolichestgvennogo opredeleniia antitel v syvorotke krovi po vzaimodeistviiu s fragmentom myshechnogo atsetilkholinovogo retseptora u bol'nykh miasteniei.

Diagnostic significance of quantitative measurement of muscle acetylcholine receptor (AChR) antibodies by ELISA in patients with myasthenia was shown. Blood serum analysis of 111 patients (30 in dynamics for various treatment types) was carried out. The most essential elevation of antibodies quantity was observed in patients with generalized myasthenia type. After thymectomy it was significantly reduced. Quantitative antibodies measurement by interaction with AChR domen in ELISA is supposed to be used as additional diagnostic method of autoimmune myasthenia.