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Microbiological evaluation of jejunal aspirates and faecal samples after oral administration of bifidobacteria and lactic acid bacteria.

A double-blind placebo controlled investigation was carried out to study the effect of peroral colonization. Human volunteers were given mixtures of bifidobacteria and lactic acid bacteria. Measurements were made over a 1 week treatment period and for another week after the end of the treatment. Two different bacteriological preparations were used, one consisted of Enterococcus faecium and Bifidobacterium longum (a total of 6.4 x 10(8) cfu d-1); the other consisted of Lactobacillus acidophilus, Bif. bifidum, Lact. delbrueckii ssp. bulgaricus, and Streptococcus thermophilus (a total of 9 x 10(9) cfu d-1). Together with a placebo preparation, they were given to 24 healthy controls (eight in each group). Microbiological examinations of jejunal aspirates showed that viable counts of most species were below the detection limit. However, the test preparation containing Ent. faecium and Bif. longum significantly reduced the anaerobe: aerobe ratio in faeces by a factor of three during treatment (P = 0.03), and increased it by a factor of 30 during the following week (P < 0.02). This study shows that peroral administration of certain bacterial cultures may affect the distal intestinal microflora.

Adult↗

Effect of inactivated salivary lysozyme on L(+)-lactic acid production in saliva and in cultures of Streptococcus mutans BHT.

The aim of this study was to evaluate the antimicrobial contribution of human lysozyme in saliva. In one series of experiments, L(+)-lactic acid (LA) production in exponential phase cultures of Streptococcus mutans BHT treated with lysozyme-deficient salivary supernatant was determined. In other experiments, LA concentration was measured in whole saliva samples from 22 school-children where the lysozyme activity had been inhibited by the addition of goat antiserum to human lysozyme (GAsL). LA production in both S. mutants cultures and saliva samples was stimulated by D-glucose addition. The results indicated a time dependent increase (approximately 30%) in LA production in lysozyme-deficient reaction-mixtures compared to untreated controls. The mean LA concentration in lysozyme-inactivated whole saliva samples was significantly higher (p less than 0.01) compared to untreated saliva. However, in 4 out of 22 children the GAsL-treatment did not affect LA production. The individual differences could not be related to salivary secretion rate, lysozyme activity or the number of S. mutans and lactobacilli in saliva. The findings of this study suggest a protective role for lysozyme in limiting acid production in saliva, but individual differences exist.

Enzyme Activation↗

Maxillary stability following Le Fort I osteotomy in combination with sagittal split ramus osteotomy and intraoral vertical ramus osteotomy: a comparative study between titanium miniplate and poly-L-lactic acid plate.

PURPOSE: The purpose of this study was to compare changes in maxillary stability after Le Fort I osteotomy with titanium miniplate and poly-L-lactic acid (PLLA) plate (Fixsorb-MX; Takiron Co, Osaka, Japan). PATIENTS AND METHODS: The subjects were composed of 47 Japanese patients with diagnosed jaw deformity: 24 underwent Le Fort I osteotomy and sagittal split ramus osteotomy (SSRO); and 23 underwent Le Fort I osteotomy intraoral vertical ramus osteotomy without internal fixation. Each group was divided into titanium plate and PLLA plate groups. Time course changes between plate groups were compared using lateral and posteroanterior cephalography. RESULTS: Significant differences were identified between titanium plate and PLLA plate groups in A point after Le Fort I osteotomy and SSRO (P < .05). Significant differences existed between titanium plate and PLLA plate groups in vertical component of posterior nasal spine after Le Fort I osteotomy in both combinations with SSRO and intraoral vertical ramus osteotomy (P < .05). However, no significant differences were identified in measurements on posteroanterior cephalography. CONCLUSION: These results suggest a slight tendency for vertical impaction after Le Fort I osteotomy both in combination with SSRO and intraoral vertical ramus osteotomy with PLLA plates, although differences in time course changes were not clinically apparent, and normal occlusion was established in all patients.

Adolescent↗

Changes in condylar long axis and skeletal stability after bilateral sagittal split ramus osteotomy with poly-L-lactic acid or titanium plate fixation.

This study was designed to assess skeletal stability after bilateral sagittal split ramus osteotomy (BSSO) and fixation with a poly-l-lactic acid (PLLA) plate, as compared to that after BSSO and fixation with a titanium plate, and to analyze the change in the condylar long axis after these procedures. The study group comprised 40 patients who had mandibular prognathism (20, titanium group; 20, PLLA group). The groups were randomized to show similar distributions of preoperative SNB. All patients underwent BSSO setback by the Obwegeser method. Fixation was done with bent titanium plates or bent PLLA plates, applied in a similar manner. Lateral, frontal, and submental-vertical cephalograms were analyzed preoperatively and postoperatively. The maximum mouth opening range and the incidence of temporomandibular disorders were also evaluated. There was no significant difference in the right condylar angle or width between the two groups, but the left condylar angle and width, gonial angle, and ramus inclination differed significantly between them (P<0.05). SNA, SNB, and ANB were similar in both groups. There was no significant difference between the groups in maximum mouth opening range or temporomandibular disorders. We conclude that the change in condylar angle after BSSO and fixation with a titanium plate is greater than that after BSSO and fixation with a PLLA plate, but skeletal stability related to the occlusion is similar for the two procedures.

Absorbable Implants↗

The tumor-preventing effect of a mixture of several lactic acid bacteria on 1,2-dimethylhydrazine-induced colon carcinogenesis in mice.

The anti-tumor effect of a dietary supplement obtained from mixed cultures of several lactic acid bacteria was examined in the colon of tumor-inducing ICR male mice by use of a carcinogen, 1,2-dimethylhydrazine (DMH, 20 mg/kg body weight, 1 intra-muscular injection per week for 10 weeks). The animals were sacrificed either 15 weeks or 24-26 weeks after the first carcinogen injection. Macroscopically, the incidence of colon tumors at a 24-26 week period of tumor induction was apparently lower in mice treated with both the DMH and dietary supplement (76%) than in those treated with DMH alone (100%). Histologically, microadenomas were induced predominantly in the anal half of the total colon, and large lymphoid aggregates were often associated with dysplastic crypts in the distal colon. Apoptotic cell masses were shed into the distended lumen of the involved crypts. The statistical analysis at a 15-week period of tumor induction indicated that the incidence of microadenomas per tumor-induced mouse was lowered significantly by use of the dietary supplement. From the present results, it is suggested that the intake of the dietary supplement inhibits the early development of colon adenomas, and the inhibition of microadenomas results in a reduction of subsequent polyp and tumor yield in the mouse colon.

1,2-Dimethylhydrazine↗

Production of interleukin 1 from macrophages incubated with poly (DL-lactic acid) granules containing ovalbumin.

The production profile of interleukin 1 (IL-1) from mouse peritoneal macrophages (M phi) was determined following their incubation with poly(DL-lactic acid) (PDLLA) granules containing ovalbumin (OVA). Upon incubation, M phi produced IL-1 at a significantly high rate compared with those incubated with OVA in the free form or OVA-free granules. A simple mixture of empty granules and free OVA exhibited the same level of IL-1 production as induced by free OVA alone. IL-1 production by the granules with a fixed OVA loading increased with an increase in their amount added to M phi. When incubated with a fixed amount of granules containing OVA of different loadings, M phi produced more IL-1 with an increase in the total OVA amount, but the IL-1 production decreased at OVA loadings higher than 10%. The presence of free OVA enhanced IL-1 production with the increased addition of empty granules, but the level induced by OVA loaded in granules was higher than that by mixtures of free OVA and empty granules, when compared at a similar OVA dose, irrespective of the absolute amount of PDLLA added. These findings indicate that the sustained release of OVA from the granules is critical to enhance the OVA-induced IL-1 production, in contrast to the OVA release accompanying a large initial burst, which reduced IL-1 production. It was concluded that the direct contact of PDLLA granules with M phi and the subsequent sustained release of OVA around M phi effectively activated M phi, resulting in enhanced IL-1 production.

Animals↗

[Study of a new lactic acid and pH 5.2 lactoserum emulsion for feminine hygiene. Results of a clinical study].

A new solution intended for use in feminine hygiene, and composed of a lactic acid base and of lactoserum at pH 5.2, was tested for a period of 8 weeks on 40 women. This test focused on its influence on the pH and the flora, as well as on the tolerance of the mucous membrane. Successive measurements of the vulvar and vaginal pH revealed no statistically significant variation between the beginning and the end of the trial. At the study inclusion thirty patients had a normal flora; of these, twenty-eight showed no change during the eight weeks of the trial. A candidiasis appeared in two patients in the middle of the trial; one of these patients received no treatment, while the other was given a five-day treatment due to similar occurrences in her past medical history. The product under study was continued throughout the trial, at the conclusion of which the flora was found to have returned to normal. At the study inclusion ten patients had either a candidiasis or a vaginitis. Their lack of symptoms and the absence of any previous problems resulted in no treatment being made in five of the ten cases, while in four cases the presence of such prior problems led to the immediate carrying out of short-length treatments. In every case, the product under study was used until the end of the trial. Bacteriological studies showed that in all the patients--whether or not they had been treated--the flora rapidly returned to normal.(ABSTRACT TRUNCATED AT 250 WORDS)

Administration, Intravaginal↗

Cultivation of fetal liver cells in a three-dimensional poly-L-lactic acid scaffold in the presence of oncostatin M.

To investigate the feasibility of fetal liver cells for liver tissue engineering, the supporting function of poly-L-lactic acid (PLLA) for fetal liver cells and the effects of oncostatin M (OSM) on hepatic differentiation were studied. After preparing three-dimensional biodegradable PLLA scaffold having a well-developed open-pore structure by a gas-forming method with ammonium chloride particles as a porogen and a gas-forming reagent, fetal liver cells separated from E14.5-C57BL/6CrSlc murine embryos were inoculated in the PLLA scaffolds. Cells were cultured in Williams' E medium with or without OSM (10 ng/ml) for 30 days with a medium change every 2 days. Results showed that there were significant increases in the number of cells and in albumin secretion in PLLA culture compared with in monolayer culture on day 15. In addition, a significant increase in albumin secretion was observed in OSM-added PLLA culture compared with OSM-free culture, and there was only a slightly enhanced albumin secretion in monolayer cultures with OSM. These results suggest that PLLA may enhance the biological activity of OSM for inducing maturation of fetal liver cells. Interestingly, the number of cells in PLLA culture with OSM decreased compared with OSM-free PLLA culture at day 15. This may be because promotion of hepatic development by OSM simultaneously suppressed in vitro hematopoiesis (i.e., blood cell production). In summary, our results indicate that the three-dimensional PLLA scaffold is a good support material for the cultivation of fetal liver cells and that OSM is capable of not only terminating hematopoiesis of the fetal liver but also stimulating the maturation of hepatic parenchymal cells in vitro.

Albumins↗

Use of rRNA gene restriction patterns to evaluate lactic acid bacterium contamination of vacuum-packaged sliced cooked whole-meat product in a meat processing plant.

Molecular typing was applied to an in-plant lactic acid bacterium (LAB) contamination analysis of a vacuum-packaged sliced cooked whole-meat product. A total of 982 LAB isolates from the raw mass, product, and the environment at different production stages were screened by restriction endonuclease (EcoRI and HindIII) analysis. rRNA gene restriction patterns were further determined for different strains obtained from each source. These patterns were used for recognizing the spoilage-causing LAB strains from the product on the sell-by day and tracing the sources and sites of spoilage LAB contamination during the manufacture. LAB typing resulted in 71 different ribotypes, of which 27 were associated with contamination routes. Raw material was distinguished as the source of the major spoilage strains. Contamination of the product surfaces after cooking was shown to be airborne. The removal of the product from the cooking forms was localized as a major site of airborne LAB contamination. Food handlers and some surfaces in contact with the product during the manufacture were also contaminated with the spoilage strains. Some LAB strains were also able to resist cooking in the core of the product bar. These strains may have an effect on the product shelf life by contaminating the slicing machine. The air in the slicing department and adjacent cold room contained very few LAB. Surface-mediated contamination was detected during the slicing and packaging stages. Food handlers also carried strains later found in the packaged product. Molecular typing provided useful information revealing the LAB contamination sources and sites of this product. The production line will be reorganized in accordance with these results to reduce spoilage LAB contamination.

Bacteria↗

Poly (D,L-lactic acid) macroporous guidance scaffolds seeded with Schwann cells genetically modified to secrete a bi-functional neurotrophin implanted in the completely transected adult rat thoracic spinal cord.

Freeze-dried poly(D,L-lactic acid) macroporous scaffold filled with a fibrin solution containing Schwann cells (SCs) lentivirally transduced to produce and secrete D15A, a bi-functional neurotrophin with brain-derived neurotrophic factor and neurotrophin-3 activity, and to express green fluorescent protein (GFP) were implanted in the completely transected adult rat thoracic spinal cord. Control rats were similarly injured and then implanted with scaffolds containing the fibrin solution with SCs lentivirally transduced to produce express GFP only or with the fibrin solution only. Transgene production and biological activity in vitro, SC survival within the scaffold in vitro and in vivo, scaffold integration, axonal regeneration and myelination, and hind limb motor function were analyzed at 1, 2, and 6 weeks after implantation. In vitro, lentivirally transduced SCs produced 87.5 ng/24 h/10(6) cells of D15A as measured by neurotrophin-3 activity in ELISA. The secreted D15A was biologically active as evidenced by its promotion of neurite outgrowth of dorsal root ganglion neurons in culture. In vitro, SCs expressing GFP were present in the scaffolds for up to 6 h, the end of a typical surgery session. Implantation of SC-seeded scaffolds caused modest loss of spinal nervous tissue. Reactive astrocytes and chondroitin sulfate glycosaminoglycans were present in spinal tissue adjacent to the scaffold. Vascularization of the scaffold was ongoing at 1 week post-implantation. There were no apparent differences in scaffold integration and blood vessel formation between groups. A decreasing number of implanted (GFP-positive) SCs were found within the scaffold during the first 3 days after implantation. Apoptosis was identified as one of the mechanisms of cell death. At 1 week and later time points after implantation, few of the implanted SCs were present in the scaffold. Neurofilament-positive axons were found in the scaffold. At 6 weeks post-grafting, myelinated axons were observed within and at the external surface of the scaffold. Axons did not grow from the scaffold into the caudal cord. All groups demonstrated a similar improvement of hind limb motor function. Our findings demonstrated that few seeded SCs survived in vivo, which could account for the modest axonal regeneration response into and across the scaffold. For the development of SC-seeded macroporous scaffolds that effectively promote axonal regeneration in the injured spinal cord, the survival and/or total number of SCs in the scaffold needs to be improved.

Animals↗

[Toxicity of cis-platinum loaded with lactic acid oligomer microspheres in mice].

We have evaluated the acute toxicity of an experimental new dosage form administered intraperitoneally in mice, consisting of cis-platinum loaded with lactic acid oligomer microspheres (CDDP-ms). The LD50 value of the CDDP-ms was 23.8 mg/kg, which amounted to 176% of the LD50 of the cis-platinum solution (13.5 mg/kg). Autopsy findings revealed no additional toxicity due to this dosage form.

Animals↗

Glucose metabolism of lactic acid bacteria changed by quinone-mediated extracellular electron transfer.

It can be expected that extracellular electron transfer to regenerate NAD+ changes the glucose metabolism of the homofermentative lactic acid bacteria. In this work, the glucose metabolism of Lactobacillusplantarum and Lactococcus lactis was examined in resting cells with 2-amino-3-carboxy-1,4-naphthoquinone (ACNQ) as the electron transfer mediator and ferricyanide (Fe(CN)6(3-)) as the extracellular electron acceptor. NADH in the cells was oxidized by ACNQ with the aid of diaphorase, and the reduced ACNQ was reoxidized with Fe(CN)6(3-). The extracellular electron transfer system promoted the generation of pyruvate, acetate, and acetoin from glucose, and restricted lactate production. Diaphorase activity increased when cultivation was aerobic, and this increased the concentrations of pyruvate, acetate, and acetoin relative to the concentration of lactate to increase in the presence of ACNQ and Fe(CN)6(3-)

Aerobiosis↗

Bacterial adherence to silver nitrate coated poly-L-lactic acid urological stents in vitro.

The purpose of this study was to see whether it is possible to prevent bacterial adherence to bioabsorbable self-reinforced L-lactic acid polymer (SR-PLLA) urological stents. The SR-PLLA stents were coated with silver nitrate blended epsilon-caprolactone/L-lactide copolymer. The adherence of five bacterial strains (Pseudomonas aeruginosa, Enterococcus faecalis, Proteus mirabilis and two strains of Escherichia coli) to coated and non-coated SR-PLLA wires were tested. It was found that silver nitrate coating prevented the adherence of bacteria (except E. faecalis) to SR-PLLA stents. The preventive effect correlated with the silver nitrate concentration. It was also found that silver nitrate coating reduced the amount of bacteria in ambient urine. In conclusion, silver nitrate coating may reduce stent-associated bacterial infections by preventing the adherence of bacteria. Further studies are needed to confirm its efficacy and safety in clinical practice.

Absorbable Implants↗

Biomimetic surface modification of poly(L-lactic acid) with chitosan and its effects on articular chondrocytes in vitro.

The objective of this study was to investigate the efficiency of two treatments for poly(L-lactic acid) (PLLA) surface modification with chitosan, via entrapment and coupling by using 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and N-hydroxysuccinimide. The properties of original PLLA films, chitosan-entrapped and coupled PLLA films were investigated by water contact angle measurement and electron spectroscopy for chemical analysis (ESCA). The contact angle indicated the change in hydrophilicity and the ESCA data suggested that the modified PLLA films became enriched with nitrogen atoms. The cytocompatibility of modified PLLA films might be improved. Therefore, the attachment and proliferation of bovine articular chondrocyte seeded on modified PLLA films and control one were examined. A whole cell enzyme-linked immunosorbent assay (Cell ELISA) that detects the BrdU incorporation during DNA synthesis and collagen type II secretion was applied to evaluate the chondrocytes on different PLLA films and tissue culture plates. Cell viability was estimated by the MTT assay and cell function were assessed by measuring sulfated glycosaminoglycan secreted by chondrocytes. These results implied that chitosan used to modify PLLA surface through entrapment and coupling could enhance the chondrocyte adhesion, proliferation and function.

Animals↗

Biomimetic surface modification of poly (L-lactic acid) with gelatin and its effects on articular chondrocytes in vitro.

Our objective in this study was to investigate the efficiency of two treatments for poly (L-lactic acid) (PLLA) surface modification with gelatin, via entrapment and coupling, using 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) and N-hydroxysuccinimide (NHS). The properties of original PLLA, gelatin-entrapped, and coupled PLLA films were investigated by water contact angle measurement and electron spectroscopy for chemical analysis (ESCA). The water contact angle indicated that the incorporation of gelatin resulted in a change in hydrophilicity, and the ESCA data suggested that the modified PLLA films became enriched with nitrogen atoms. The cytocompatibility of modified PLLA films might be improved. Therefore, we examined the attachment and proliferation of bovine articular chondrocyte seeded on modified PLLA films and virgin films. A whole-cell enzyme-linked immunosorbent assay (cell ELISA) that detects 5-bromo-2'-deoxyuridine (BrdU) incorporation during DNA synthesis and collagen type II secretion was applied to evaluate the chondrocytes on different PLLA films and tissue culture plates (TCPS). Cell viability was estimated by the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] assay, and cell function was assessed by measuring glycosaminoglycan (GAG) secreted by chondrocytes. These results implied that gelatin used to modify the PLLA surface through entrapment and coupling could enhance chondrocyte adhesion, proliferation, and function.

Animals↗

Bioactive poly(L-lactic acid) conduits seeded with Schwann cells for peripheral nerve regeneration.

This study attempted to enhance the efficacy of peripheral nerve regeneration using our previously tested poly(L-lactic acid) (PLLA) conduits by incorporating them with allogeneic Schwann cells (SCs). The SCs were harvested, cultured to obtain confluent monolayers and two concentrations (1 x 10(4) and 1 x 10(6) SC/ml) were combined with a collagen matrix (Vitrogen) and injected into the PLLA conduits. The conduits were then implanted into a 12 mm right sciatic nerve defect in rats. Three control groups were used: isografts, PLLA conduits filled with collagen alone and empty silicone tubes. The sciatic functional index (SFI) was calculated monthly through four months. At the end of second and fourth months, the gastrocnemius muscle was harvested and weighed for comparison and the graft conduit and distal nerve were harvested for histomorphologic analysis. The mean SFI demonstrated no group differences from isograft control. By four months, there was no significant difference in gastrocnemius muscle weight between the experimental groups compared to isograft controls. At four months, the distal nerve demonstrated a statistically lower number of axons mm2 for the high and low SC density groups and collagen control. The nerve fiber density was significantly lower in all of the groups compared to isograft controls by four months. The development of a "bioactive" nerve conduit using tissue engineering to replace autogenous nerve grafts offers a potential approach to improved patient care. Although equivalent nerve regeneration to autografts was not achieved, this study provides promising results for further investigation.

Animals↗

Polysaccharide surface engineering of poly(D, L-lactic acid) via electrostatic self-assembly technique and its effects on osteoblast growth behaviours.

The objective of this study was to surface modify the poly (D, L-lactic acid) (PDLLA) films and assess the effects of the modified surfaces on the functions of osteoblasts cultured in vitro. A layer-by-layer (LBL) self assembly technique, was used leading to the formation of multilayers on the PDLLA film surfaces. Chitosan (Chi) and poly (styrene sulfonate, sodium salt) (PSS) were utilized as polycation and polyanion in this study, respectively. The layer structure was investigated by using X-ray photoelectron spectroscopy (XPS) and water contact angle measurement, respectively. XPS analysis displayed the presence of chitosan on PDLLA surface. A full coverage of coating with PSS/Chi layers was achieved on the PDLLA surface only after the deposition layers of PEI/(PSS/Chi)2. These results showed that PDLLA films could be modified with PSS/Chi pairs which may affect the biocompatibility of the modified PDLLA films. To confirm this hypothesis, cell proliferation, cell viability as well as alkaline phosphtase activity of osteoblasts on layer-by-layer modified PDLLA films as well as control samples were investigated in vitro. The proliferation of osteoblasts on modified PDLLA films was found to be greater than that on control (p < 0.05 and p < 0.01) after 1, 4 and 7 days culture, respectively. Cell viability measurement showed that the PSS/Chi modified PDLLA films have higher cell viability (p < 0.01) than control. Osteoblast differentiation function (ALP) on LBL-modified PDLLA film was found significantly higher (p < 0.01) than that of virgin PDLLA films. These data suggests that PSS/Chi pair was successfully employed to surface modify PDLLA film via a layer-by-layer technique, and enhanced its cell biocompatibility.

Animals↗