Adherence of bacteria to mucus collected from different parts of the reproductive tract of heifers and cows.

In the present study, we examined the adherence of indigenous vaginal bacteria, probiotic strains, and metritis pathogens to mucus collected from different parts of the reproductive tracts of heifers and cows and compared their adherence with the bacterial adherence to mucus collected from the stomach and large intestine of pigs. Most of the vaginal strains adhered to mucus collected from different parts of the reproductive tract and strongly adhered to gastric mucus, with the exception of Lactobacillus buchneri 24S8. Only Lactobacillus mucosae 29S8, Enterococcus faecium E21, and E. faecium EAC adhered to colonic mucus. Probiotic strains adhered strongly to mucus collected from the reproductive tract and gastric mucus but did not adhere to colonic mucus. Pathogenic strains were adherent to vaginal, uterine horn, and gastric mucus, except Escherichia coli O8:K88ab:H9 (65), Fusobacterium necrophorum, and Gardnerella vaginalis, which adhered to uterine cervix mucus. Only Kocuria kristinae and G. vaginalis adhered to uterine body mucus; E. coli O149:K88ac (EC) adhered to colonic mucus. The strains did not exhibit host specificity but rather strain specificity. The ability to adhere to mucus was a characteristic unique to each strain. To our knowledge, this is the first report regarding in vitro adherence of GRAS (Generally Regarded As Safe) lactobacilli isolated from different sources to mucus collected from different parts of the reproductive tract.

Effects of the administration of lactobacilli, maltodextrins and fructooligosaccharides upon the adhesion of E. coli O8:K88 to the intestinal mucosa and organic acid levels in the gut contents of piglets.

The influence of the administration of Lactobacillus plantarum, maltodextrin Maldex 150 and Raftifeed IPX fructooligosaccharides on the inhibition of adhesion of E. coli O8:K88 to the mucosa of the jejunum, ileum and colon as well as on the organic acid levels was investigated in 33 conventional piglets. The counts of E. coli K88 adhering to the jejunal mucosa were significantly decreased (p < 0.05) in Lact. plantarum + Maldex 150 and Lact. plantarum + Maldex 150 + Raftifeed IPX groups. The counts of E. coli K88 adhering to the colonic mucosa of Lact. plantarum + Maldex 150 + Raftifeed IPX and Lact. plantarum + Raftifeed IPX groups were significantly lower (p < 0.05) than in Lact. plantarum and Lact. plantarum + Maldex 150 animals. The acetic acid levels in the ileum and colon of the Lact. plantarum + Maldex 150 + Raftifeed IPX group and Lact. plantarum + Raftifeed IPX group were significantly higher (p < 0.05) than in the Lact. plantarum and Lact. plantarum + Maldex 150 group. The combination of Lact. plantarum, maltodextrin Maldex 150 and Raftifeed IPX proved to be the most effective one to inhibit the counts of E. coli O8:K88 adhering to the intestinal mucosa of the jejunum and colon of conventional piglets.

Reduction of Salmonella in gnotobiotic Japanese quails caused by the enterocin A-producing EK13 strain of Enterococcus faecium.

The effect of the enterocin A-producing EK strain of Enterococcus faecium on Salmonella dusseldorf SA31 was tested in gnotobiotic Japanese quails. Sixteen 3-day-old gnotobiotic Japanese quails, hatched from disinfected eggs placed in sterile boxes, were divided into two groups of 8 birds: a control group, which was inoculated orally with the SA31 strain (1 x 10(7) CFU/ml), and the experimental group, which was inoculated orally with 200 microl of E. faecium (1 x 10(9) CFU/ml), 16 h before infection with the S. dusseldorf. The latter group then received the same average dose of E. faecium daily in drinking water. Faecal samples were taken 8, 24, 48 and 168 h after the inoculation of S. dusseldorf and examined for S. dusseldorf and E. faecium (EK13). The quails were then killed and the number of the EK13 strain of E. faecium and of S. dusseldorf in the caecum and ileum were estimated. A reducing effect of the EK13 strain against the SA31 strain in faeces was detected in the samples taken at 24 and 48 h from the group with the EK13 strain. Significant reductions were also found in the numbers of S. dusseldorf SA31 strain in the caecum but not in the ileum.

Improvement of the probiotic effect of micro-organisms by their combination with maltodextrins, fructo-oligosaccharides and polyunsaturated fatty acids.

Probiotics could represent an effective alternative to the use of synthetic substances in nutrition and medicine. The data concerning the efficacy of probiotics are often contradictory. This paper focuses on the enhancement of the efficacy of probiotics by their combination with synergistically acting components of natural origin. Maltodextrins can be obtained by enzymatic hydrolysis of starch and are suitable for consumption. Administration of Lactobacillus paracasei together with maltodextrin decreased the number of Escherichia coli colonising the jejunal mucosa of gnotobiotic piglets by 1 logarithm compared to the control group. Fructo-oligosaccharides (FOS) are naturally occurring oligosaccharides, mainly of plant origin. L. paracasei administered in combination with FOS significantly increased counts of Lactobacillus spp., Bifidobacterium spp., total anaerobes and total aerobes compared to the control group as well as the L. paracasei group. It also significantly decreased Clostridium and Enterobacterium counts in the faeces of the weanling piglets compared with the control group. Dietary lipids influence the gastrointestinal microbiota and specifically the population of lactic acid bacteria. In gnotobiotic piglets the oral administration of an oil containing polyunsaturated fatty acids (PUFA) significantly increased the number of L. paracasei adhering to jejunal mucosa compared to the control group. Our results showed that maltodextrin KMS X-70 and PUFA can be used to enhance the effect of probiotic micro-organisms in the small intestine, and similarly FOS enhance the effect of probiotic micro-organisms in the large intestine.

Results of animal experiments using an undulation pump total artificial heart: analysis of 10 day and 19 day survival.

An undulation pump is a special rotary blood pump in which rotation of a brushless DC motor is transformed to an undulating motion by a disc in the pump housing attached by means of a special link mechanism. In the blood pump, a closed line between the disc and housing moves from the inlet to the outlet by this undulating disc motion, which sucks and pushes the blood from the inlet to the outlet. Because the same phenomena occurs at both sides of the disc, a continuous flow is obtained when the motor rotational speed is constant. The pump flow pattern can be easily changed from continuous flow to pulsatile flow by controlling the motor drive current pattern. A seal membrane made of segmented polyurethane protects the blood from invading the link mechanism as well as the motor. UPTAH is fabricated with two undulation pumps and two brushless DC motors. Its size is 75 mm in diameter and 80 mm long, and it has one of the great advantage of no compliance chamber required in the system. UPTAHs were implanted under cardiopulmonary bypass (CPB) into the chest cavities of 16 goats, each weighing between 41 and 72 kg. No anticoagulant and antiplatelet agent was used after the surgery. The left atrial pressure was automatically controlled to prevent its elevation and sucking of the atrial wall into the atrial cuff. The following results were obtained: (1) UPTAHs fit well into all the goats; (2) the longest survival was 19.8 days, the cause of death was bleeding from the aortic anastomosis; (3) No thrombus was observed in the blood pump despite no anticoagulant use. Hemolysis depended upon the length of CPB during surgery. When CPB time was within 2 hours, hemolysis level returned to baseline within a few days of the surgery. UPTAH is a promising implantable TAH, because of its small size and easy controllability.

Implantation of the undulation pump total artificial heart in the goat.

The undulation pump total artificial heart (UPTAH) was developed by using small-size continuous-flow displacement-type blood pumps (undulation pump). To clarify and improve the problems accompanied by the implantation in the chest, 14 animal experiments were performed on goats weighing 41.3-79.2 kg. The UPTAH could be implanted in the chest of all goats and was driven with a modulation pulsatile mode. The first problem was the atrial suction effect. This problem could be prevented to some extent by developing the soft disk and by improving atrial cuffs. An automatic detection and releasing of the atrial suction effect was also tried. The next problem was acute lung edema accompanied by the postural change of the animal. Development of the automatic control of left atrial pressure could prevent this problem. Small blood leakage from a pinpoint hole in the seal membrane was the next problem. Improvement of the manufacturing procedure of the membrane prevented this. With these improvements, a 10 day survival could be obtained with this unique implantable total artificial heart.

Study of the effect of Lactobacillus paracasei and fructooligosaccharides on the faecal microflora in weanling piglets.

The influence of administration of Lactobacillus paracasei alone and mixture of Lactobacillus paracasei and fructooligosaccharide on faecal bacteria counts in the weanling pigs was investigated. The administration of Lactobacillus paracasei alone significantly decreased Clostridium (p < 0.05) and Enterobacteriaceae (p < 0.05) counts as compared to the control. Lactobacillus paracasei administered in combination with fructooligosaccharide significantly increased Lactobacillus (p < 0.01-p < 0.05), Bifidobacterium (p < 0.05), total anaerobes (p < 0.05), and total aerobes (p < 0.05) counts compared to control group as well as Lactobacillus paracasei group and significantly decreased Clostridium (p < 0.05) and Enterobacteriaceae (p < 0.01) counts compared to control group. The results obtained point out to a synergic effect of the combination of Lactobacillus paracasei and fructooligosaccharide on numbers of bacterial populations observed in the faeces of the weanling pigs.

Present status of the total artificial heart at the University of Tokyo.

At the University of Tokyo, various types of total artificial heart (TAH) systems have been studied since 1959. At the present time, 2 types of implantable TAH have been developed. One is an undulation pump TAH (UPTAH) and the other is a flow transformed pulsatile TAH (FTPTAH). Using the UPTAH, 14 cases of implantation were performed in goats and 10 days' survival obtained. The new type of FTPTAH is under a prototype study. To prevent ring thrombus, a polyurethane membrane valve, a jellyfish valve, has been developed. The longest in vivo experiences with this valve in the TAH blood pump have been 312 days in the left side blood pump and 414 days in the right side blood pump. Conductance and arterial pressure based control (1/R control) can realize the physiological control of the TAH. Using 1/R control, 532 days of survival could be obtained in a goat with a paracorporeal TAH. The technique required to apply this control method to a implantable TAH is under development. We have proposed a new 5 year research project of the implantable TAH entitled "Comprehensive Basic Research on the Development of a Japanese Original Implantable Total Artificial Heart" to The Ministry of Welfare.

In vitro improvement of bone marrow-derived hematopoietic colony formation in HIV-positive patients by alpha-D-tocopherol and erythropoietin.

The majority of patients with progressive HIV infection develop a severe hematopoietic failure which is aggravated by the hematotoxic effect of azidothymidine (AZT) treatment. Since it was shown in a mouse model that alpha-D-tocopherol (vitamin E derivative) antagonizes the inhibitory influence of AZT on the growth of burst-forming units-erythrocyte (BFU-E), it was the aim of this study to investigate whether alpha-D-tocopherol and high dosages of erythropoietin (EPO) increase the hematopoietic colony-forming capacity of bone marrow cells from patients with progressive HIV disease and especially if they reverse the inhibitory effects of AZT. The data demonstrate that tocopherol (1-100 mumol/l) significantly increases the growth of BFU-E and colony-forming units granulocyte-monocyte (CFU-GM) from HIV-infected patients. This stimulatory effect is dose-dependent (maximum at 30-100 mumol/l) and only occurs when the agent is present from the beginning of the cultures. EPO (5-10 U/ml) also augments the numbers of BFU-E from HIV-infected patients. Tocopherol equally ameliorates the growth of BFU-E and CFU-GM from the HIV-positive cohort in the presence of AZT (10-100 mumol/l). For healthy controls, no such increase was observed, either with tocopherol or with higher dosages of EPO. In conclusion, both tocopherol and EPO partially reverse the myelosuppressive action of AZT in HIV-positive patients.

Evaluation study of calves with total artificial heart (TAH) surviving for 218-293 days of pumping.

Evaluation of five longest surviving calves among a total of 54 longterm survivors, at the Brno Research Centre was performed. These calves were treated by administration of antihypertensives and partially by electrical stimulation of the right atrium to reduce central venous pressure (CVP). Treatment for CVP was effective, as compared with two groups of 5 calves each not treated in this way, and it is assumed to be an important factor for prolongation of survival. Mineralization of driving diaphragms was present in all these calves. No preventive measures of this pathologic change were performed in this group. Another complication is hemolysis, which is partly caused by immune mechanisms initiated by administration of alpha-methyl dopa and partly by mechanical destruction due to a heavily mineralized driving diaphragm. The prevention of mineralization is another important problem for future research. Other problems, concerning the survival of TAH recipients are discussed.

Recent efforts in artificial heart research in Czechoslovakia.

The authors present basic information about the technical development of the total artificial heart (TAH) and TAH drivers. Long-term experiments, in which the TAH TNS-Brno-II and VII and the Rostock hearts were implanted, are described. Long-term experiments were dedicated to analysis of specific pathophysiologic problems (venous hypertension, thrombogenesis, and calcification) in order to increase survival rates. Thrombogenesis has been partially solved in the TNS-BRNO-VII device by optimal TAH construction using an asymmetric driving diaphragm, the undulating motion of which produced excellent results. Two methods for prevention of increase in central venous pressure (CVP) are verified: administration of antihypertensive drugs and atrial electrical stimulation. Of 155 calves studied, 50 long-term experiments in calves and one in a goat were done. The survival of these animals extended from 31 to 293 days of pumping (average survival, 116.9 days). To prevent driving diaphragm calcification, we have incorporated exogenic inhibitors of calcification into the diaphragm biomaterial (polyurethane). A unique long-term experiment with intrathoracic TAH implantation in the goat is also described. In addition clinical use of the TAH as a bridge to transplantation is discussed. Parallel research with the left ventricular assist device (LVAD) system was studied experimentally, and twice used in patients. The TAH TNS-BRNO-VII/80/clin/ was used in four patients. It worked faultlessly for 22 hours to 10 days. More precise and specific indications for TAH implantation when used as a bridge to transplantation are needed.

[Therapy of venous hypertension in animals with an artificial heart and long-term survival]. / K terapii zilní hypertenze u zvírat dlouhodobe prezívajících s totální náhradou srdce.

1. The authors elaborated an original concept for the interpretation of vasomotor disorders in subjects with an artificial heart. 2. This concept is based on the regeneration of nervous elements in the walls of the atria (in particular the right one) after implantation of the artificial heart and on comparison of their activity with the venous pressure which revealed the interrelationship of the two phenomena. 3. Both therapeutic methods based on this concept, the method of influencing the afferentation and efferentation of vasomotor nervous regulations leading to a reduction of the central venous pressure proved valid and effective. 4. Evaluation of the effectiveness of this therapy is based on regular assessment of the central venous pressure, on the laboratory assay of enzymes (AST, ALT, GMT and LDH), on the assessment of serum albumin and finally on the morphological and histological examination of the liver incl. assessment of the hepatic index. 5. The nervous pathogenesis is closely linked with hormonal factors. The latter are conceived as the participation of associated factors in the pathogenesis of venous hypertension in recipients with an artificial heart.

Therapy of venous hypertension in calves with total artificial heart (TAH).

Vasomotor disregulation, preponderantly expressed by a pathological increase of central venous pressure (CVP) in calves with total artificial heart (TAH), starts to be evident from about the 50th day of pumping. The main cause of this state is an imbalance in cardiac receptor areas. Ventricular vasodepressor mechanisms are eliminated with the ventricular tissue, which is replaced by the artificial blood pump. In the stumps of both atria, which remain in situ, all neural elements disappear immediately after TAH implantation, but within two months they are fully regenerated. Regenerated atrial receptors are the starting points of afferent neural stimuli, which in the vasomotor center of the brainstem, increase the activity of the vasoconstricting functional component. A general tendency to vasoconstriction, now not well counterbalanced, increases, and the progressive venous hypertension causes loss of liver function and morphology. Two therapeutic approaches were tried: afferent therapy by atrial electrical stimulation, and efferent therapy by the administration of antihypertensives. Both kinds of this therapy were sufficiently effective in reducing CVP, protecting the liver, and prolonging average survival.

Serum aldosterone studies in 10 calves living for 2-5 months with a total artificial heart (TAH).

A group of 10 experimental calves that lived for 61-147 days with a TAH was used to study serum aldosterone levels by RIA with ALDOCTK-125 CEA Kit (Sorin Biomedica, Italy). The aldosterone levels were plotted against the serum sodium and potassium, body weight, CVP, urinary sodium and potassium and with hematocrit levels. In these experiments, three types of TAH (TNS-BRNO II, VII, and Rostock) were used. The levels of aldosterone were already high before the surgery and during the entire pumping period, which indicates the development of secondary aldosteronism. The study deals with the mechanisms that participate in the development of aldosteronism in each phase of the experiment with the TAH.

Surgical aspects of the implantation of a left ventricle assist device.

The authors have summarised their experience with short-term use of a left ventricle assist device (LVAD). Nine experiments were carried out in healthy calves. A polyurethane pump of their own design, placed extracorporeally, was used. Its in-flow cannula was placed in the left atrium, the outflow one in the initial segment of the descending aorta. The LVAD was operated for a period of 5 days. Fourteen days after its disconnection, the animal was sacrificed and both the bypass system and parenchymal organs were examined. No surgical complications were observed during the experiments. Macro- and microscopical pictures showed minor infarctions in the renal tissue which, however, did not impair renal function. The findings in other organs were related to anaesthesia, the operative procedure and way of animal sacrificing. The experiments have proved the safety of the bypass system used over a short period and, consequently, its potential for future clinical application.

Control and driving of pneumatic total artificial hearts TNS-BRNO-II and -III in long-term experiments.

Hemodynamic analysis was carried out during long-term experiments with the pneumatic total artificial hearts TNS-BRNO-II and TNS-BRNO-III to determine standard methods of starting artificial hearts and criteria for their long-term operation in vivo. In long-term experiments, regulatory mechanisms automatically regulating the systole length and diastolic aspiration pressure have also been verified. Comparison of hemodynamic variables obtained from invasive measurements with pneumatic pressure curves permitted the control and monitoring of the experiment in its entirety by noninvasive methods only. The control of the artificial heart using the Chirasist TN 3 and Chirasist TN 4 was adapted to specific properties of the pumps, above all to the functions of the atypical inlet valves. The terminal stages of the experiments have shown that a 100-ml pump can ensure survival of experimental calves up to 210 kg body weight.

Hematological and biochemical studies in calves living over 100 days with the polymethylmethacrylate total artificial heart TNS Brno II.

A polymethylmethacrylate total artificial heart (kinetic components made of polyetherurethane) of TNS Brno II type was implanted into seven calves (2-5 months of age) surviving for the average of 152.4 +/- 19.1 days after the implantation. During the entire post-operative period the animals received oral warfarin-sodium, acetylsalicylic acid, dipyridamole and alpha-tocopherol. Blood was taken for biochemical and hematological examinations twice a week from the jugular vein. During the experiments there were decreases in the number of red blood cells, hematocrit and hemoglobin levels. Plasma free hemoglobin and serum enzymes (alkaline phosphatase, AST, ALT, LDH) increased. Coagulation tests were abnormal because anticoagulation therapy was used. There were minimal changes in the number of white blood cells and platelets, fibrinogen, blood pH, blood glucose, serum electrolytes, bilirubin (total and direct), creatinine, blood urea, and lactate. Possible reasons for observed changes include the gradual rise in the central venous pressure and damaged function of the liver parenchyma. Other factors playing a possible role in inducing changes in laboratory findings are also discussed.