Comparative effectiveness of faecal microbiota transplant by route of administration.

The optimal route of delivery for faecal microbiota transplant (FMT) is unknown. This observational single-centre study analysed the two-week cure rates for all patients who received FMT from 2013 to 2016 according to route of delivery. Overall, nasogastric delivery of FMT was less effective than lower endoscopic delivery. When patients were stratified by illness severity, nasogastric delivery achieved similar cure rates in healthier individuals, whereas lower endoscopic delivery was preferred for relatively ill individuals. Nasogastric delivery may be less effective than lower endoscopic delivery; however, when taking the cost, preparation and potential risk into account, this difference may not be clinically significant for patients with mild disease.

Proteolytic cleavage of carboxypeptidase N markedly increases its antifibrinolytic activity.

BACKGROUND: Carboxypeptidase N (CPN) is a constitutively active basic carboxypeptidase sharing specificity with activated thrombin-activable fibrinolysis inhibitor (TAFIa). Generally, CPN is regarded as being non-antifibrinolytic. However, this assumption has not been thoroughly investigated, particularly with respect to long-term antifibrinolysis. In addition, a recent report has shown that plasmin cleavage increases the catalytic activity of CPN. Therefore, we investigated the antifibrinolytic properties of CPN and plasmin-cleaved CPN (CPNc). METHODS: CPN was incubated with plasmin for various periods of time and the prolongation of clot lysis at various concentrations of CPN/CPNc mixture was investigated in TAFI-depleted plasma. CPN cleavage was analyzed by electrophoresis and catalytic activity was determined by monitoring cleavage of the small substrate, FA-Ala-Lys. RESULTS: CPN exhibited antifibrinolytic properties in plasma clot lysis assays when present at supraphysiological concentrations. Depletion of CPN from plasma decreased the lysis time of clots formed from minimally diluted plasma at low tissue-type plasminogen activator (t-PA) concentrations. Plasmin cleavage of CPN markedly increased the antifibrinolytic properties. CPN and CPNc prolonged lysis in a non-saturable, dose-dependent, and t-PA-dependent manner. At sufficient concentration, CPN and CPNc prolonged lysis at least forty-fivefold. CPNc was 700% more antifibrinolytic than CPN but only 7% more active toward FA-Ala-Lys. The active site inhibitor GEMSA eliminated the antifibrinolytic effects of CPN and CPNc. Antifibrinolytic activity correlated with cleavage of active and/or regulatory subunits, presumably generating heterodimeric CPNc. CONCLUSIONS: Limited proteolysis of CPN by plasmin generates an enzyme with greatly increased antifibrinolytic properties. We speculate that (patho)physiological proteolysis of CPN may generate a long-term antifibrinolytic enzyme.

Complete inhibition of fibrinolysis by sustained carboxypeptidase B activity: the role and requirement of plasmin inhibitors.

BACKGROUND: The antifibrinolytic effect of activated thrombin-activatable fibrinolysis inhibitor (TAFIa) and carboxypeptidase B (CPB) displays threshold behavior. When CPB was used to simulate conditions mimicking continuous TAFIa activity, it affected the lysis of plasma clots differently to clots formed from a minimal fibrinolytic system comprising fibrinogen, plasminogen and alpha(2)-antiplasmin. Whereas CPB saturably prolonged clot lysis in the purified system, the effect of CPB did not appear saturable in plasma clots. METHODS: To rationalize this difference, we investigated the effects of alpha(2)-antiplasmin, alpha(2)-macroglobulin, antithrombin and aprotinin on CPB-mediated antifibrinolysis. RESULTS: CPB alone prolonged fibrinolysis in a saturable manner and the efficacy of CPB increased with decreasing tissue-type plasminogen activator (t-PA) concentration. The inhibitors by themselves did not halt fibrinolysis and the potency of each inhibitor in the absence of CPB mirrored their solution-phase plasmin inhibitory potentials: alpha(2)-antiplasmin approximately equal to aprotinin >> alpha(2)-macroglobulin >> antithrombin. With both CPB and inhibitor present, a synergistic effect was observed. The antifibrinolytic sensitivity to CPB was related to the plasmin inhibitory potential of the inhibitor. CONCLUSIONS: Fibrinolysis could be completely inhibited by alpha(2)-antiplasmin, alpha(2)-macroglobulin and antithrombin, but not aprotinin, in the presence of CPB, and occurred only when the irreversible inhibitor or pool of inhibitors were in excess of plasminogen. Western blot analysis indicated that the CPB-mediated shutdown of fibrinolysis was a result of plasminogen consumption prior to clot lysis. The CPB concentration required for fibrinolytic shutdown was dependent on t-PA concentration and the inhibitory potential of the irreversible inhibitor pool.

Efficacy of single-dose intravenous dolasetron versus ondansetron in the prevention of postoperative nausea and vomiting.

BACKGROUND: Postoperative nausea and vomiting (PONV) is a significant problem in surgical patients. The 5-hydroxytryptamine3-receptor antagonists ondansetron, dolasetron, and granisetron are being used to prevent PONV and avoid the adverse events associated with traditional antiemetics such as antihistaminic agents, anticholinergic agents, and dopamine antagonists. OBJECTIVE: Because practitioners have taken widely differing approaches to the selection and dosing of agents in this class, this retrospective study assessed the relative efficacy of i.v. dolasetron and ondansetron in preventing PONV when used according to their approved labeling. METHODS: The medical charts of patients who underwent total abdominal hysterectomy or laparoscopic cholecystectomy and received either dolasetron 12.5 mg or ondansetron 4 mg were reviewed. Efficacy was assessed based on the number of episodes of PONV and time to the occurrence of PONV in the 24 hours after surgery. RESULTS: Of 75 medical records reviewed, 59 met the criteria for inclusion in the efficacy analysis. There were no statistically significant between-group differences in demographic or baseline clinical characteristics. The majority of patients were obese (body mass index > or = 27 kg/m2), had no history of either PONV or motion sickness, and underwent total abdominal hysterectomy. PONV occurred in 11 of 25 (44%) patients receiving dolasetron and 18 of 34 (53%) patients receiving ondansetron. Four patients receiving dolasetron experienced PONV in the first 2 hours after surgery, compared with 7 patients receiving ondansetron. CONCLUSION: There were no significant differences in efficacy between single doses of i.v. dolasetron 12.5 mg and i.v. ondansetron 4 mg in the prevention of PONV.

NSAID impairment of orthodontic tooth movement.

OBJECTIVE: To evaluate the impairment of orthodontic tooth movement caused by nonsteroidal antiinflammatory drugs (NSAIDs). DATA SOURCES: Biomedical literature accessed through MEDLINE (1966-January 2000), EMBASE (1980-January 2000), and International Pharmaceutical Abstracts (1970-January 2000). Key search terms included NSAIDs, orthodontics, and tooth movement. DATA SYNTHESIS: Orthodontic dentistry applies mechanical force to generate tooth movement. Since prostaglandins are mediators of tooth movement, it is reasonable to expect that prostaglandin inhibitors, such as NSAIDs, inhibit or delay tooth movement. An evaluation of studies measuring the extent of NSAID impairment on tooth movement was undertaken. CONCLUSIONS: Results from animal studies have shown that NSAIDs can impair the tooth movement process. Until long-term human data are obtained, acetaminophen remains an appropriate alternative to NSAIDs for treating orthodontic-associated pain.

A kinetic analysis of the tissue plasminogen activator and DSPAalpha1 cofactor activities of untreated and TAFIa-treated soluble fibrin degradation products of varying size.

The kinetics of tissue plasminogen activator (t-PA) and DSPAalpha1-catalyzed plasminogen activation using untreated and TAFIa-treated fibrin degradation products (FDPs), ranging in weight average molecular weight (M(w)) from 0.48 x 10(6) to 4.94 x 10(6) g/mol, were modeled according to the steady-state template model. The FDPs served as effective cofactors for both activators. The intrinsic catalytic efficiencies of both t-PA (17.4 x 10(5) m(-1) s(-1)) and DSPAalpha1 (6.0 x 10(5) m(-1) s(-1)) were independent of FDP M(w). The intrinsic catalytic efficiency of t-PA was 12-fold higher than that measured under identical conditions with intact fibrin as the cofactor. At sub-saturating levels of cofactor and substrate, rates were strongly dependent on FDP M(w) with DSPAalpha1 but not t-PA. Loss of activity with decreasing FDP M(w) correlated with loss of finger-dependent binding of the activators to the FDPs. TAFIa treatment of the FDPs resulted in 90- and 215-fold decreases in the catalytic efficiencies of t-PA (0.20 x 10(5) m(-)(1) s(-1)) and DSPAalpha1 (0.028 x 10(5) m(-1) s(-1)), yielding cofactors that were still 30- and 50-fold better than fibrinogen with t-PA and DSPAalpha1, respectively. Our results show that for both activators the products released during fibrinolysis are very effective cofactors for plasminogen activation, and both t-PA and DSPAalpha1 cofactor activity are strongly down-regulated by TAFIa.

Proteolytic processing of human coagulation factor IX by plasmin.

Previous studies have shown that thrombin generation in vivo caused a 92% decrease in factor IX (F.IX) activity and the appearance of a cleavage product after immunoblotting that comigrated with activated F.IX (F.IXa). Under these conditions, the fibrinolytic system was clearly activated, suggesting plasmin may have altered F.IX. Thus, the effect(s) of plasmin on human F.IX was determined in vitro. Plasmin (50 nM) decreased the 1-stage clotting activity of F.IX (4 microM) by 80% and the activity of F.IXa (4 microM) by 50% after 30 minutes at 37 degrees C. Plasmin hydrolysis of F.IX yields products of 45, 30, 20, and 14 kd on reducing sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and 2 products of 52 and 14 kd under nonreducing conditions. Plasmin-treated F.IX did not bind the active site probe, p-aminobenzamidine, or form an SDS-stable complex with antithrombin. It only marginally activated human factor X in the presence of phospholipid and activated factor VIII. Although dansyl-Glu-Gly-Arg-chloromethyl ketone inactivated-F. IXa inhibited the clotting activity of F.IXa, plasmin-treated F.IX did not. Plasmin cleaves F.IX after Lys43, Arg145, Arg180, Lys316, and Arg318, but F.IXa is not appreciably generated despite cleavage at the 2 normal activation sites (Arg145 and Arg180). Tissue plasminogen activator-catalyzed lysis of fibrin formed in human plasma results in generation of the 45- and 30-kd fragments of F.IX and decreased F.IX clotting activity. Collectively, the results suggest that plasmin is able to down-regulate coagulation by inactivating F.IX.

Parasites of domestic and wild animals in South Africa. XXXVIII. Ixodid ticks collected from 23 wild carnivore species.

Ixodid ticks were collected from 104 wild carnivores belonging to 23 species in various nature reserves and on several farms in all nine provinces of South Africa. Seven feral cats in a nature reserve were also examined. Twenty-four tick species belonging to seven genera were recovered and identified. Amongst these ticks we consider the adults of Haemaphysalis leachi, Haemaphysalis spinulosa, Haemaphysalis zumpti, Ixodes rubicundus, Rhipicentor nuttalli, Rhipicephalus simus and Rhipicephalus turanicus to be true parasites of wild carnivores. Although numerous adult Rhipicephalus appendiculatus and Rhipicephalus zambeziensis were collected from some lions these were either sick or old animals. The immature stages of seven species regularly utilized wild carnivores as hosts on an opportunistic basis.

The molecular weights, mass distribution, chain composition, and structure of soluble fibrin degradation products released from a fibrin clot perfused with plasmin.

We used a perfused clot system to study the degradation of cross-linked fibrin. Multiangle laser light scattering showed that plasmin-mediated cleavage caused the release of noncovalently associated fibrin degradation products (FDPs) with a weight-averaged molar mass (Mw) of approximately 6 x 10(6) g/mol. The Mw of FDPs is dependent on ionic strength, and the Mw observed at 0.15 M NaCl resulted from the self-association of FDPs having Mw of approximately 3.8 x 10(6) g/mol. Complete solubilization required the cleavage of approximately 25% of fragment D/fragment E connections, with 48% alpha-, 62% beta-, and 42% gamma-chains cleaved. These results showed that D-E cleavage cannot be explained by a random mechanism, implying cooperativity. Gel filtration and multiangle laser light scattering showed that FDPs range from 2.5 x 10(5) to 1 x 10(7) g/mol. In addition to fragment E, FDPs are composed of fragments ranging from 2 x 10(5) Da (D-dimer, or DD) to at least 2.3 x 10(6) Da (DX8D). FDP mass distribution is consistent with a model whereby FDPs bind to fibrin with affinities proportional to fragment mass. Root mean square radius analysis showed that small FDPs approximate rigid rods, but this relationship breaks down as FDPs size increases, suggesting that large FDPs possess significant flexibility.

A study of the mechanism of inhibition of fibrinolysis by activated thrombin-activable fibrinolysis inhibitor.

TAFI (thrombin-activable fibrinolysis inhibitor) is a recently described plasma zymogen that, when exposed to the thrombin-thrombomodulin complex, is converted by proteolysis at Arg92 to a basic carboxypeptidase that inhibits fibrinolysis (TAFIa). The studies described here were undertaken to elucidate the molecular basis for the inhibition of fibrinolysis. When TAFIa is included in a clot undergoing fibrinolysis induced by tissue plasminogen activator and plasminogen, the time to achieve lysis is prolonged, and free arginine and lysine are released over time. In addition, TAFIa prevents a 2.5-fold increase in the rate constant for plasminogen activation which occurs when fibrin is modified by plasmin in the early course of fibrin degradation. The effect is specific for the Glu- form of plasminogen. TAFIa prevents or at least attenuates positive feedback expressed through Lys-plasminogen formation during the process of fibrinolysis initiated by tissue plasminogen activator and plasminogen. TAFIa also inhibits plasmin activity in a clot and prolongs fibrinolysis initiated with plasmin. We conclude that TAFIa suppresses fibrinolysis by removing COOH-terminal lysine and arginine residues from fibrin, thereby reducing its cofactor functions in both plasminogen activation and the positive feedback conversion of Glu-plasminogen to Lys-plasminogen. At relatively elevated concentrations, it also directly inhibits plasmin.

Effects of the neuraminidase inhibitor zanamavir on otologic manifestations of experimental human influenza.

Middle ear pressure (MEP) abnormalities are frequently observed during influenza virus infection and may serve as surrogate markers for the risk of otitis media. MEP abnormalities were evaluated in adult volunteers who were inoculated with influenza A/Texas/36/91(H1N1) or B/Yamagata/88 virus and given the antiviral zanamivir (GG167) intranasally as prophylaxis or early treatment in randomized, double-blind, placebo-controlled trials. In the influenza A prophylaxis studies, 15% of 61 zanamivir recipients versus 61% of 33 placebo recipients showed significant MEP abnormalities (P < .01). In the influenza A early treatment trial, 32% of 31 infected zanamivir recipients versus 73% of 26 infected placebo recipients developed MEP abnormalities (P < .01). In the influenza B prophylaxis trial, 16% of 25 zanamivir versus 44% of 9 placebo recipients showed abnormalities (P = .09). These findings indicate that the neuraminidase inhibitor zanamivir, which is effective in reducing experimental influenza illness, provides protection against the development of MEP abnormalities.

The solution phase interaction between apolipoprotein(a) and plasminogen inhibits the binding of plasminogen to a plasmin-modified fibrinogen surface.

In the present study, we assessed the binding of recombinant forms of apolipoprotein(a) [r-apo(a)] to plasminogen. Apo(a)-plasminogen interactions were demonstrated to be lysine-dependent, as they were abolished by the addition of epsilon-aminocaproic acid. Binding of r-apo(a) and plasma-derived Lp(a) to Glu-plasminogen was assessed in solution using a mutant form of recombinant plasminogen [Plg(S741C)] labeled at the active site with 5'-(iodoacetamido)fluorescein. High-affinity binding of apo(a) to plasminogen was observed with the 17-kringle r-apo(a) (Kd = 20.1 +/- 3.3 nM) as well as with plasma-derived Lp(a) (Kd = 5.58 +/- 0.08 nM). Binding studies using various truncated and mutant forms of r-apo(a) demonstrated that sequences within apo(a) kringle IV types 2-9 and the strong lysine binding site (LBS) in apo(a) kringle IV type 10 are not required for high-affinity binding to plasminogen. In all cases, the binding stoichiometry for the apo(a)-plasminogen interaction was determined to be 1:1. Binding data obtained using a 17-kringle r-apo(a) derivative lacking the protease-like domain (17KDeltaP; Kd = 3158 +/- 138 nM) indicate that sequences within the protease-like domain of apo(a) mediate its interaction with LBS in plasminogen. We determined that r-apo(a) and plasminogen bind to distinct sites on plasmin-modified fibrinogen with the concentration of plasminogen binding sites exceeding the concentration of r-apo(a) sites by a factor of 10. Furthermore, r-apo(a) is capable of inhibiting the binding of plasminogen to plasmin-modified fibrinogen surfaces, an effect which we show is attributable to the formation of a solution phase apo(a)/plasminogen complex which exhibits a greatly reduced affinity for plasminogen binding sites on plasmin-modified fibrinogen. The results of this study provide new insights into the mechanism by which apo(a) and Lp(a) may inhibit fibrinolysis, thus contributing to the atherothrombotic risk associated with this lipoprotein.

Rhipicephalus interventus sp. nov. (Acari: Ixodidae), a new tick species closely related to Rhipicephalus tricuspis Dönitz, 1906 and Rhipicephalus Iunulatus Neumann, 1907, from east and central Africa.

Descriptions of the adults of this new species are given, together with information on its hosts and distribution. Previously it was referred to briefly by Walker, Keirans, pegram & Clifford (1988), who noted that in many respects it is intermediate in appearance between R. tricuspis and R. Iunulatus.

Enzymatic synthesis of aminocyclitol moieties of aminoglycoside antibiotics from inositol by Streptomyces spp.: detection of glutamine-aminocyclitol aminotransferase and diaminocyclitol aminotransferase activities in a spectinomycin producer.

Extracts of stationary-phase mycelia of the spectinomycin producer Streptomyces flavopersicus ATCC 19756 catalyzed inositol dehydrogenase, L-glutamine:inosose aminotransferase, 2-epi-streptamine:inosose aminotransferase, streptamine:inosose aminotransferase, N3-methyl-2-deoxystreptamine:inosose aminotransferase, and aminodeoxy-scyllo-inositol:inosose aminotransferase reactions, as detected with a new rapid assay procedure. These results suggest that one or both amino groups of the N1,N3-dimethyl-2-epi-streptamine moiety of spectinomycin are derived by transamination from the alpha-amino group of L-glutamine. An enzymatic procedure for distinguishing among N1- and N3-monomethyl diaminocyclitol derivatives and their diaminocyclitol biosynthetic precursors is described. A scheme showing key roles of glutamine-aminocyclitol aminotransferases in biosynthesis of major aminoglycoside antibiotics is presented.

Quality assurance of ultrasound imaging instruments by monitoring the monitor.

Ultrasound quality assurance (QA) is a means of assuring the constant performance of an ultrasound instrument. A novel 'ultrasound image analyser' has been developed to allow objective, accurate and repeatable measurement of the image displayed on the ultrasound screen, i.e. as seen by the operator. The analyser uses a television camera/framestore combination to digitize and analyse this image. A QA scheme is described along with the procedures necessary to obtain a repeatable measurement of the image so that comparisons with earlier good images can be made. These include repositioning the camera and resetting the video display characteristics. The advantages of using the analyser over other methods are discussed. It is concluded that the analyser has distinct advantages over subjective image assessment methods and will be a valuable addition to current ultrasound QA programmes.

GM-1 ganglioside administration combined with physical therapy restores ambulation in humans with chronic spinal cord injury.

In a randomized double-blind cross-over study, humans with chronic spinal cord injury received ganglioside GM-1 or placebo for 2 months. GM-1, administered intravenously at a dose of 100 mg, 6 days a week, resulted in a statistically significant improvement of motor scores (P < 0.05), whether administered before or after 2 months of placebo. There was no placebo effect on motor scores. Subjects who received GM-1 before placebo maintained their improvement during the placebo phase. Subjects who received GM-1 ambulated with a reciprocal gait, using orthotics, for longer distances and at a faster rate whether the drug was administered before or after placebo. These results constitute the first finding that any chemical substance improves locomotion in human chronic spinal cord injury.