Simple faecal preparation and efficacy of frozen inoculum in faecal microbiota transplantation for recurrent Clostridium difficile infection--an observational cohort study.

BACKGROUND: Faecal microbiota transplantation (FMT) is an effective treatment for recurrent Clostridium difficile infection (rCDI). The finding of suitable donor, donor screening and preparation of faecal transplants are challenging in clinical work. AIM: To develop a practical protocol for preparing frozen transplants and to compare the efficacy of previously frozen and fresh faeces in treating rCDI. METHODS: Two healthy volunteers acted as universal donors for the frozen faecal preparations, which were prepared by suspending faeces into physiological saline, adding glycerol to a final concentration of 10% and storing at -80 °C. We compared the outcomes of patients with rCDI who had undergone FMT at colonoscopy and received infusion of previously prepared, freeze-stored faeces (n = 23) or fresh faeces from individual (n = 15) or universal donors (n = 11) (total n = 49). Clinical failure was defined as persistent or recurrent symptoms with a positive C. difficile toxin stool test, and a need for new therapy. RESULTS: At 12 weeks post-FMT, symptoms were resolved in 22 of 23 patients receiving previously frozen faeces, and in all 11 or 14 of 15 patients receiving fresh faeces from the universal or individual donors respectively (totally 25 of 26; P = ns, success rate 96%). Mild transient fever appeared for two patients receiving frozen faeces, but no other significant side effects were observed. 42 patients were followed up for a year post-FMT and the success rate was 88% in both fresh and frozen faeces groups. CONCLUSIONS: Preparation of frozen transplants simplifies the practical aspects of faecal microbiota transplantation without loss of efficacy or safety.

The zygomatic bone as a potential donor site for alveolar reconstruction--a quantitative anatomic cadaver study.

The aim of this cadaver study was to evaluate the possibility of using the zygomatic bone as an intraoral bone harvesting donor site and to determine the safety of this harvesting procedure. In addition, the volume of bone material harvested from the zygomatic bone was measured. Twenty fixed adult cadavers were used to yield a total of 40 zygomatic bone harvest sites, from which bone was collected. The volume of bone obtained from the zygomatic harvests was measured with a water displacement method and by compressing the graft into a syringe. The safety of the technique was evaluated by assessing possible encroachment upon the neighbouring structures. After bone harvesting, the zygomatic sites were exposed and evaluated for visible perforations or fractures. Possible damage to the neighbouring tissues was also examined with computed tomography scans at 18 sites in nine cadavers. The average bone graft volume obtained from the zygomatic bone was measured to be 0.53 ml (SD 0.25) with water displacement and 0.59 ml (SD 0.26) with the syringe. The complications in the zygoma included 15 small perforations into the maxillary sinus and 7 perforations into the infratemporal fossa. CT scans showed that bone could be harvested safely without encroaching upon the orbital floor or the surrounding nerves and vessels in the zygoma. The zygomatic bone is a safe intraoral donor site for the reconstruction of small- to medium-sized alveolar defects.

Tuberous sclerosis: clinicopathologic features and review of the literature.

INTRODUCTION: Tuberous sclerosis is a hamartoneoplastic syndrome, which may involve multiple organ systems. Oral hard tissue manifestations of the syndrome have been described in the literature only as recently as 1955. Patients who presented with clinical manifestations of tuberous sclerosis did not routinely undergo oral surveys to rule out 'lesions', and consequently data on 'lesions' in the maxillofacial complex is scant. Ten cases have been found in the English language literature, which describe maxillofacial 'lesions', which may be tumours, new growths, neoplasms or overgrowths occurring in patients diagnosed with tuberous sclerosis. PURPOSE: To review the literature for all maxillofacial lesions associated with tuberous sclerosis and to present an eleventh case of a patient with a maxillofacial lesion diagnosed as having tuberous sclerosis. RESULTS: Eleven cases were found with maxillofacial fibroblastic lesions associated with tuberous sclerosis. These lesions were all fibrous benign neoplasms found in the maxillofacial bony complex. CONCLUSIONS: Maxillofacial fibroblastic lesions in tuberous sclerosis have various histopathological presentations, some of which may be difficult to differentiate. Consequently, close microscopic examination of these lesions is necessary so that adequate surgical treatment is provided.

A natural ErbB4 isoform that does not activate phosphoinositide 3-kinase mediates proliferation but not survival or chemotaxis.

ErbB4 is a member of the epidermal growth factor receptor (ErbB) family that mediates cellular responses activated by neuregulins (NRG) and other epidermal growth factor-like growth factors. Two naturally occurring ErbB4 isoforms, ErbB4 CYT-1 and ErbB4 CYT-2, have previously been identified. Unlike ErbB4 CYT-1, ErbB4 CYT-2 lacks a phosphoinositide 3-kinase (PI3-K)-binding site and is incapable of activating PI3-K. We have now examined the consequences of the inability of this isoform to activate PI3-K on cell proliferation, survival, and chemotaxis in response to NRG-1beta: (i) NRG-1beta stimulated proliferation of cells expressing either ErbB4 CYT-1 or ErbB4 CYT-2. Consistent with the mitogenic responsiveness, analysis of downstream signaling showed that Shc and MAPK were phosphorylated after stimulating either isoform with NRG-1beta. (ii) NRG-1beta protected cells expressing ErbB4 CYT-1 but not cells expressing ErbB4 CYT-2 from starvation-induced apoptosis as measured by effects on cell number and 4', 6-diamidino-2-phenylindole staining. Furthermore, in cells expressing ErbB4 CYT-2, Akt, a protein kinase that mediates cell survival, was not phosphorylated. (iii) NRG-1beta stimulated chemotaxis and membrane ruffling in cells expressing ErbB4 CYT-1 but not in cells expressing ErbB4 CYT-2. In summary, ErbB4 CYT-2 can mediate proliferation but not chemotaxis or survival. These results suggest a novel mechanism by which cellular responses such as chemotaxis and survival may be regulated by the expression of alternative receptor-tyrosine kinase isoforms that differ in their coupling to PI3-K signaling.

Comparison of four bone collectors designed for oral and maxillofacial surgery--an in vitro study.

Four bone collectors designed for oral and maxillofacial surgery were compared in an in vitro study. Two commercially available bone collectors (Osseus Coagulum Trap, Quality Aspirators and Frios, Friatec) with mesh sizes of 0.17 x 0.17 and 0.44 x 2.40 mm, respectively and models developed by the authors with mesh sizes of 0.355 x 0.355 or 0.50 x 0.50 (Model I) and 0.80 x 0.80 (Model II) were tested. All collectors are designed to be connected to the suction device. Five dissected pig mandibles were used to determine the harvesting capacity during drilling and suctioning for 30 s. All collectors harvested bone quite effectively, but the highest amount of bone chips was harvested by Model I which had been designed by the investigators.

Physiological degradation converts the soluble syndecan-1 ectodomain from an inhibitor to a potent activator of FGF-2.

The activity of fibroblast growth factor 2 (FGF-2) is stringently controlled. Inactive in undisturbed tissues, it is activated during injury and is critical for tissue repair. We find that this control can be imposed by the soluble syndecan-1 ectodomain, a heparan sulfate proteoglycan shed from cell surfaces into wound fluids. The ectodomain potently inhibits heparin-mediated FGF-2 mitogenicity because of the poorly sulfated domains in its heparin sulfate chains. Degradation of these regions by platelet heparanase produces heparin-like heparin sulfate fragments that markedly activate FGF-2 mitogenicity and are found in wound fluids. These results establish a novel physiological control for FGF-2 and suggest new ways to modulate FGF activity.

Syndecans, heparan sulfate proteoglycans, maintain the proteolytic balance of acute wound fluids.

An imbalance between proteases and antiproteases is thought to play a role in the inflammatory injury that regulates wound healing. The activities of some proteases and antiproteases found in inflammatory fluids can be modified in vitro by heparin, a mast cell-derived glycosaminoglycan. Because syndecans, a family of cell surface heparan sulfate proteoglycans, are the major cellular source of heparin-like glycosaminoglycan, we asked whether syndecans modify protease activities in vivo. Syndecan-1 and syndecan-4 ectodomains are shed into acute human dermal wound fluids (Subramanian, S. V., Fitzgerald, M. L., and Bernfield, M. (1997) J. Biol. Chem. 272, 14713-14720). Moreover, purified syndecan-1 ectodomain binds cathepsin G (Kd = 56 nM) and elastase (Kd = 35 nM) tightly and reduces the affinity of these proteases for their physiological inhibitors. Purified syndecan-1 ectodomain protects cathepsin G from inhibition by alpha1-antichymotrypsin and squamous cell carcinoma antigen 2 and elastase from inhibition by alpha1-proteinase inhibitor by decreasing second order rate constants for protease-antiprotease associations (kass) by 3700-, 32-, and 60-fold, respectively. Both enzymatic degradation of heparan sulfate and immunodepletion of the syndecan-1 and -4 in wound fluid reduce these proteolytic activities in the fluid, indicating that the proteases in the wound environment are regulated by interactions with syndecan ectodomains. Thus, syndecans are shed into acute wound fluids, where they can modify the proteolytic balance of the fluid. This suggests a novel physiological role for these soluble heparan sulfate proteoglycans.

Information of circulation from soft tissue and dental pulp by means of pulsatile reflected light: further development of optical pulp vitalometry.

OBJECTIVES: To show experimentally and clinically that information of blood circulation patterns can be obtained through dentin and enamel by means of reflected light. STUDY DESIGN: Several different experimental techniques were developed in vitro leading up to in vivo tests. RESULTS: Pulsatile information of back-scattered light at 560 nm was obtained in an experiment in which a thread with white and red sections was pulled through the empty pulp cavum. The pulsatile information of back-scattered light of 560 nm wavelength was detected from a forefinger of a volunteer through dental hard tissue plates. There was a difference in the radiant flux of back-scattered light in extracted teeth filled with various materials. Photoplethysmograms obtained from the vital pulp of a 24-year-old woman were analyzed with Fast Fourier Transformation (FFT). The analyzed data showed a dominant amplitude of heart rate near 1 Hz. CONCLUSIONS: Tests indicated that reliable information of circulation can be obtained from the dental pulp chamber by means of the optical reflection method. This could serve as a model for a new type of pulp vitalometer.

Expression of small extracellular chondroitin/dermatan sulfate proteoglycans is differentially regulated in human endothelial cells.

We have examined the expression of the small extracellular chondroitin/dermatan sulfate proteoglycans (CS/DS PGs), biglycan, decorin, and PG-100, which is the proteoglycan form of colony stimulating factor-1, in the human endothelial cell line EA.hy 926. We have also examined whether modulation of the phenotype of EA.hy 926 cells by tumor necrosis factor-alpha (TNF-alpha) is associated with specific changes in the synthesis of these PGs. We demonstrate that EA.hy 926 cells, when they form monolayer cultures typical of macrovascular endothelial cells, express and synthesize detectable amounts of biglycan and PG-100, but not decorin. On SDS-polyacrylamide gel electrophoresis both PGs behave like proteins of the relative molecular weight of approximately 250,000. TNF-alpha that changed the morphology of the cells from a polygonal shape into a spindle shape and that also stimulated the detachment of the cells from culture dish, markedly decreased the net synthesis of biglycan, whereas the net synthesis of PG-100 was increased. These changes were parallel with those observed at the mRNA level of the corresponding PGs. The proportions of the different sulfated CS/DS disaccharide units of PGs were not affected by TNF-alpha. Several other growth factors/cytokines, such as interferon-gamma, fibroblast growth factors-2 (FGF-2) and -7 (FGF-7), interleukin-1beta, and transforming growth factor-beta, unlike TNF-alpha, modulated neither the morphology nor the biglycan expression of EA.hy 926 cells under the conditions used in the experiments. However, PG-100 expression was increased also in response to FGF-2 and -7 and transforming growth factor-beta. None of the above cytokines, including TNF-alpha, was able to induce decorin expression in the cells. Our results indicate that the regulatory elements controlling the expression of the small extracellular CS/DS PGs in human endothelial cells are different.

Factors related to Periotest values in endosseal implants: a 9-year follow-up.

Periotest values (PTV) of successful endosseal implants of 2 one-stage implant systems. TPS and ITI, were followed from 3 months to 9 years in order to determine the factors that contribute to the values. 128 TPS screw implants were inserted in the lower jaw of 34 subjects, (mean age 55 years), for retaining overdentures. 108 ITI implants were inserted in the upper and lower jaws in 50 subjects (mean age 42 years), for retaining overdentures, crowns and bridges. PTVs were first measured after the osseointegration period and thereafter annually. First of all there was a difference between the 2 implant systems. Mean PTVs of TPS bicortical screws were significantly lower (P < 0.05) than those of ITI implants (screws, hollow-screws, hollow-cylinders). Factors which significantly contributed to PTVs of ITI implants were jaw (upper/lower), implant length and region of the jaw in which the implant was inserted. PTVs of ITI implants in the lower jaw were lower than in the upper jaw (P < 0.05). The length of implant had no effect on PTVs in the lower jaw, but in the upper jaw, PTVs of ITI 8-10-mm implants were lower than 12-mm implants (P < 0.05). PTVs of implants inserted in the anterior region of the upper jaw were higher than those in the posterior region (P < 0.05). In conclusion, bone quality and implant length had a statistically significant effect on implant mobility in long-term follow-up. PTVs of various implant systems, however, differ, a fact that must be taken into account in evaluating the success of implants.

A method of harvesting corticocancellous bone chips for reconstructive maxillofacial surgery.

A method is described to harvest bone chips for maxillofacial surgery with large burs and a special collecting net connected to a suction device. Three nets and four burs were tested in vitro on a pig mandible to determine the optimal combination. All burs produced bone chips of rather similar sizes. The midsized net (hole diameter 0.5 mm) collected most bone chips during a 1-min drilling.

Suppression of syndecan-1 expression in endothelial cells by tumor necrosis factor-alpha.

Syndecan-1 is a cell surface proteoglycan that binds extracellular matrix components and modulates the activity of heparin-binding growth factors. The expression of syndecan-1 is modified during development, carcinogenesis, and tissue regeneration. During cutaneous wound healing, syndecan-1 expression is transiently induced in newly-formed capillaries of granulation tissue as well as in proliferating keratinocytes. To study the mechanisms underlying this regulation we investigated the effects of several growth factors/cytokines on syndecan-1 expression in two human cell lines: EA.hy 926 endothelial cells and HaCaT keratinocytes. None of these factors significantly altered syndecan-1 mRNA expression in cultured keratinocytes, but when given to endothelial cells, tumor necrosis factor-alpha (TNF-alpha) specifically and dose-dependently suppressed syndecan-1 expression at both mRNA and protein levels. TNF-alpha reduced the amount of syndecan-1 protein in EA.hy 926 cells in both the presence and absence of serum and, at the same time, induced the expression of intercellular adhesion molecule-1 (ICAM-1). The suppressive effect of TNF-alpha on endothelial syndecan-1 expression was reproducible in in vivo experiments in which TNF-alpha-coated beads were administered directly to healing skin wounds of mice. Data supporting these findings were further obtained by injecting TNF-alpha into an experimental rat granulation tissue model. In this tissue TNF-alpha suppressed syndecan-1 mRNA expression by approximately 80%. These results indicate that TNF-alpha is capable of down-regulating syndecan-1 expression in endothelial cells both in vitro and in vivo and suggest that similar mechanisms may be responsible for the changes in syndecan-1 expression observed during various regenerative, developmental, and malignant processes.