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investigation of the impact of feeding lactobacillus plantarum crl 1815 encapsulated in microbially derived polymers on the rat faecal microbiota.the aim of this study was to evaluate the impact of the administration of microencapsulated lactobacillus plantarum crl 1815 with two combinations of microbially derived polysaccharides, xanthan : gellan gum (1%:0·75%) and jamilan : gellan gum (1%:1%), on the rat faecal microbiota.201222612542
inhibition of bacterial carbonic anhydrases and zinc proteases: from orphantargets to innovative new antibiotic drugs.zinc-containing enzymes, such as carbonic anhydrases (cas) and metalloproteases (mps) play critical functions in bacteria, being involved in various steps of their life cycle, which are important for survival, colonization, acquisition of nutrients for growth and proliferation, facilitation of dissemination, invasion and pathogenicity. the development of resistance to many classes of clinically used antibiotics emphasizes the need of new antibacterial drug targets to be explored. there is a weal ...201222214451
distinct gut microbiota in southeastern african and northern european infants.the intestinal microbiota composition in infants reflects the early environment. our objective was to compare the gut microbiota in 6-month-old infants living in rural malawi with children of the same age living in urban finland, both being breast-fed and having an age-appropriate diet typical for each area.201222228076
enhancement of the structural stability of full-length clostridial collagenase by calcium ions.the clostridial collagenases g and h are multidomain proteins. for collagen digestion, the domain arrangement is likely to play an important role in collagen binding and hydrolysis. in this study, the full-length collagenase h protein from clostridium histolyticum was expressed in escherichia coli and purified. the n-terminal amino acid of the purified protein was ala31. the expressed protein showed enzymatic activity against azocoll as a substrate. to investigate the role of ca(2+) in providing ...201222685155
nonsurgical treatment of dupuytren's contracture: 1-year us post-marketing safety data for collagenase clostridium histolyticum.collagenase clostridium histolyticum (cch) is a food and drug administration-approved treatment for adult patients with dupuytren's contracture with a palpable cord that has been shown efficacious and safe in clinical trials.201223730231
treatment for chemotherapy-induced alopecia in mice using parathyroid hormone agonists and antagonists linked to a collagen binding domain.parathyroid hormone (pth) agonists and antagonists have been shown to improve hair growth after chemotherapy; however, rapid clearance and systemic side-effects complicate their usage. to facilitate delivery and retention to skin, we fused pth agonists and antagonists to the collagen binding domain (cbd) of clostridium histolyticum collagenase. in-vitro studies showed that the agonist fusion protein, pth-cbd, bound collagen and activated the pth/parathyroid hormone-related peptide receptor in sa ...201122130912
Temporal and spatial distribution of Bacillus and Clostridium histolyticum in swine manure composting by fluorescent in situ hybridization (FISH).The temporal and spatial distribution of the genus Bacillus and Clostridium histolyticum group in swine manure composting was determined by fluorescent in situ hybridization using fluorescently labeled 16S rRNA-targeted oligonucleotide probes LGC353b and Chis150, respectively. The temporal distribution of total bacteria, Bacillus and C. histolyticum, detected in each layer of the composting pile was noticeable in that the number of them detected at the high-temperature stage was higher than that ...201121881890
polycystic kidney disease-like domains of clostridial collagenases and their role in collagen recruitment.bacterial collagenases exhibit a multimodular domain organization. while the n-terminal collagenase unit harbors the catalytic zinc and suffices to degrade peptidic substrates, collagen substrates come in different types, explaining the requirement for accessory domains such as polycystic kidney disease (pkd)-like domains for efficient catalysis. how the recognition and unfolding of (micro-)fibrillar or triple-helical collagen is accomplished are only poorly understood. here, we present the crys ...201121871007
In vitro fermentation by human gut bacteria of proteolytically digested caseinomacropeptide nonenzymatically glycosylated with prebiotic carbohydrates.The in vitro fermentation selectivity of hydrolyzed caseinomacropeptide (CMP) glycosylated, via Maillard reaction (MR), with lactulose, galacto-oligosaccharides from lactose (GOSLa), and galacto-oligosaccharides from lactulose (GOSLu) was evaluated, using pH-controlled small-scale batch cultures at 37 °C under anaerobic conditions with human feces. After 10 and 24 h of fermentation, neoglyconjugates exerted a bifidogenic activity, similar to those of the corresponding prebiotic carbohydrates. No ...201122004447
▾xiapex for dupuytren's contracture.dupuytren's contracture is a benign, progressive disabling condition of the hands. traditionally, surgery (e.g. fasciotomy, fasciectomy, joint fusion and amputation) has been the only effective treatment but it is not an option for every patient. a potential alternative intervention is a formulation of collagenase clostridium histolyticum (▾xiapex--pfizer limited), which has been licensed for injection directly into the affected joint as treatment for adults with dupuytren's contracture.5 here w ...201122170410
Structure of collagenase G reveals a chew-and-digest mechanism of bacterial collagenolysis.Collagen constitutes one-third of body protein in humans, reflecting its extensive role in health and disease. Of similar importance, therefore, are the idiosyncratic proteases that have evolved for collagen remodeling. The most efficient collagenases are those that enable clostridial bacteria to colonize their host tissues; but despite intense study, the structural and mechanistic basis of these enzymes has remained elusive. Here we present the crystal structure of collagenase G from Clostridiu ...201121947205
specific prebiotics in a formula for infants with phenylketonuria.this exploratory study investigated the influence of adding a patented, specific mixture of prebiotic oligosaccharides (scgos/lcfos [9:1 ratio], danone research) to a protein substitute suitable for infants with phenylketonuria (pku); pku anamix infant (nutricia).201121967856
The impact of perinatal probiotic intervention on gut microbiota: Double-blind placebo-controlled trials in Finland and Germany.Specific probiotic combinations during early feeding, via the mother or incorporated in early formula-feeding, mold the intestinal microbiota composition in infants. The objective was to analyze the impact of probiotic administration to mother or infant on gut microbiota composition in 6-month-old Finnish and German infants. In Finland probiotics were given to mothers (n = 79) for 2 months prior to and 2 months after delivery. In Germany probiotics were started in infants (n = 81) at weaning, at ...201121979491
Effects of pH and ORP on microbial ecology and kinetics for hydrogen production in continuously dark fermentation.The microbial structure and kinetic characteristics of the hydrogen producing strains in two fermentative continuous stirred-tank reactors (CSTRs) were studied by controlling pH and oxidation and reduction potential (ORP). The fluorescence in situ hybridization (FISH) tests were conducted to investigate the fermentative performance of Clostridium histolyticum (C. histolyticum), Clostridium lituseburense (C. lituseburense) and Enterobacteriaceae. The experimental results showed that in ethanol-ty ...201121978625
effect of hesperidin in vitro on root dentine collagen and demineralization.caries progress might be controlled when collagen matrix could be preserved after demineralization. the aim of this ph cycling study was to investigate the effect of hesperidin, a citrus flavonoid antioxidant, on dentine collagen and remineralization in dentine lesion, and compared with that of chlorhexidine.201121414383
specific prebiotics modulate gut microbiota and immune activation in haart-naive hiv-infected adults: results of the "copa" pilot randomized trial.intestinal mucosal immune system is an early target for human immunodeficiency virus type 1 (hiv-1) infection, resulting in cd4(+) t-cell depletion, deterioration of gut lining, and fecal microbiota composition. we evaluated the effects of a prebiotic oligosaccharide mixture in highly active antiretroviral therapy (haart)-naive hiv-1-infected adults. in a pilot double-blind, randomized, placebo-controlled study, 57 haart-naive hiv-1-infected patients received a unique oligosaccharide mixture (15 ...201121525866
Dupuytren Disease: An Evolving Understanding of an Age-old Disease.Dupuytren disease, a clinical entity originally described more than 400 years ago, is a progressive disease of genetic origin. Excessive myofibroblast proliferation and altered collagen matrix composition lead to thickened and contracted palmar fascia; the resultant digital flexion contractures may severely limit function. The pathophysiology is multifactorial and remains a topic of research and debate. Genetic predisposition, trauma, inflammatory response, ischemia, and environment, as well as ...201122134207
Collagenase clostridium histolyticum injection for the treatment of Dupuytren's contracture.Collagenase clostridium histolyticum is a novel treatment for Dupuytren's contracture, approved by the U.S. Food and Drug Administration in February 2010. Prior to its availability, surgery was the only treatment for contracture related to this disorder. Dupuytren's disease is a benign, progressive fibroproliferative disorder affecting the palms of the hands. It is characterized by the formation of collagen- rich nodules and cords, which gradually shorten by the action of myofibroblasts, resulti ...201121971540
the risk to human islet cell transplant recipients of acquiring variant creutzfeldt-jakob disease: a provisional quantitative risk assessment. 201121691192
the treatment of dupuytren disease.the treatment of progressive dupuytren contractures has historically been and continues to be largely surgical. although a number of surgical interventions do exist, limited palmar fasciectomy continues to be the most common and widely accepted treatment option. until recently, nonsurgical options were limited and clinically ineffective. however, the commercial availability and recent approval of collagenase clostridium histolyticum now provides practitioners with a nonsurgical approach to this ...201121527148
unfolding bent fingers: new handiwork for bacteria. 201121322908
molecular mechanochemistry: low force switch slows enzymatic cleavage of human type i collagen monomer.in vertebrate animals, fibrillar collagen accumulates, organizes, and persists in structures which resist mechanical force. this antidissipative behavior is possibly due to a mechanochemical force-switch which converts collagen from enzyme-susceptible to enzyme-resistant. degradation experiments on native tissue and reconstituted fibrils suggest that collagen/enzyme kinetics favor the retention of loaded collagen. we used a massively parallel, single molecule, mechanochemical reaction assay to d ...201121348512
protective effect of withania somnifera and cardiospermum halicacabum extracts against collagenolytic degradation of collagen.the irreversible destruction of extracellular matrix (ecm) such as cartilage, tendon, and bone that comprise synovial joints is the hallmark of both rheumatoid arthritis and osteoarthritis by over-expression of matrix metalloproteinase (mmp)-collagenases. we report herein the detailed study on the inhibitory effects of withania somnifera extract (wse) and cardiospermum halicacabum extract (che) on clostridium histolyticum collagenase (chc) activity against the degradation of the ecm component of ...201121789568
monthly administration of a novel pth-collagen binding domain fusion protein is anabolic in mice.we synthesized fusion proteins of parathyroid hormone (pth) (1-33) and the collagen binding domain of colh (cbd) and tested them for anabolic bone activity in mice. two fusion proteins were synthesized, linking the carboxy terminus of pth(1-33) either directly to the amino terminal of the cbd or to the cbd through an adjacent colh domain (pth-pkd-cbd). both pth-cbd and pth-pkd-cbd increased camp accumulation in cells stably transfected with the pth/pthrp receptor, and both peptides bound to type ...201121512758
tissue dissociation enzymes for isolating human islets for transplantation: factors to consider in setting enzyme acceptance criteria.tissue dissociation enzymes are critical reagents that affect the yield and quality of human pancreatic islets required for islet transplantation. the united states food and drug administration's oversight of this procedure recommends laboratories to set acceptance criteria for enzymes used in the manufacture of islet products for transplantation. currently, many laboratories base this selection on personal experience because biochemical analysis is not predictive of success of the islet isolati ...201121116222
studies on collagen-tannic acid-collagenase ternary system: inhibition of collagenase against collagenolytic degradation of extracellular matrix component of collagen.we report the detailed studies on the inhibitory effect of tannic acid (ta) on clostridium histolyticum collagenase (chc) activity against degradation of extracellular matrix component of collagen. the ta treated collagen exhibited 64% resistance against collagenolytic hydrolysis by chc, whereas direct interaction of ta with chc exhibited 99% inhibition against degradation of collagen and the inhibition was found to be concentration dependant. the kinetic inhibition of chc has been deduced from ...201121823836
cloning of a novel collagenase gene from the gram-negative bacterium grimontia (vibrio) hollisae 1706b and its efficient expression in brevibacillus choshinensis.the collagenase gene was cloned from grimontia (vibrio) hollisae 1706b, and its complete nucleotide sequence was determined. nucleotide sequencing showed that the open reading frame was 2,301 bp in length and encoded an 84-kda protein of 767 amino acid residues. the deduced amino acid sequence contains a putative signal sequence and a zinc metalloprotease consensus sequence, the hexxh motif. g. hollisae collagenase showed 60 and 59% amino acid sequence identities to vibrio parahaemolyticus and v ...201121515782
collagenase clostridium histolyticum. short-term benefit in dupuytren's contracture, unknown long-term risks.dupuytren's contracture is due to fibroblast overgrowth in the aponeuroses of the hand and fingers, leading to excessive collagen production. this can cause functional impairment due to retraction of one or several fingers.the standard treatment is surgery, but relapses are frequent. a product containing collagenolytic bacterial enzymes (collagenases) is now marketed for local treatment of these retractions. two double-blind placebo-controlled trials in 308 and 66 patients show that 1 to 3 local ...201122216538
in vitro fermentation and prebiotic potential of novel low molecular weight polysaccharides derived from agar and alginate seaweeds.fermentation properties and prebiotic potential of novel low molecular weight polysaccharides (lmwps) derived from agar and alginate bearing seaweeds was investigated. ten lmwps were supplemented to ph, temperature controlled anaerobic batch cultures inoculated with human feces from three donors, in triplicate. microbiota changes were monitored using fluorescent in-situ hybridization and short chain fatty acids, the fermentation end products were analysed using gas chromatography. of the ten lmw ...201121924371
Characterization and functional assessment of Clostridium histolyticum class I (C1) collagenases and the synergistic degradation of native collagen in enzyme mixtures containing class II (C2) collagenase.Clostridium histolyticum expresses two classes of collagenases, C1 and C2. However, degradation of these enzymes by proteases during the fermentation or purification process may lead to numerous molecular forms that lead to inconsistent release of islets from human pancreata. This report defines the amino acid sequence of the truncated forms of C1 (C1b or C1c) that contain a single collagen-binding domain (CBD) and investigates the synergy between the different forms of C1 collagenase and C2 to ...201122099748
In vitro fermentation of alternansucrase raffinose-derived oligosaccharides by human gut bacteria.In this work, in vitro fermentation of alternansucrase raffinose-derived oligosaccharides, previously fractionated according to their degree of polymerization (DP; from DP4 to DP10), was carried out using small-scale pH-controlled batch cultures at 37 °C under anaerobic conditions with human feces. Bifidogenic activity of oligosaccharides with DP4-6 similar to that of lactulose was observed; however, in general, a significant growth of lactic acid bacteria Bacteroides , Atopobium cluster, and ...201121913653
[collagenase clostridum histolyticum in the management of dupuytren's contracture].dupuytren's contracture is a fibroproliferate disease of the palmar aponeurosis with a formation of nodules and cords. surgical treatment is the gold standard for dupuytren's contracture at the moment. a short while ago collagenase clostridium histolyticum was licensed as a non-surgical method to treat dupuytren's contracture. collagenase clostridium histolyticum is injected directly into the dupuytren's cord and after 24 h the contracture is distended by manual rupturing. collagenase clostridiu ...201121935843
determining clinically important changes in range of motion in patients with dupuytren's contracture: secondary analysis of the randomized, double-blind, placebo-controlled cord i study.injectable collagenase clostridium histolyticum is efficacious in correcting dupuytren's contracture as assessed by changes in the angle of contracture and range of motion (rom). however, clinically important changes in rom have not been evaluated in depth. the objective of this secondary analysis of the cord i trial was to identify severity levels using baseline rom, estimate a clinically important difference (cid) for rom, and link the results to collagenase treatment and patient satisfaction.201121967070
ph in the bacteria-contaminated wound and its impact on clostridium histolyticum collagenase activity: implications for the use of collagenase wound debridement agents.we sought to understand the influence of ph on clostridium collagenase activity, using both in vitro and in vivo, and to understand the influence of bacterial contamination on ph in vivo.201121860332
high molecular weight coffee melanoidins are inhibitors for matrix metalloproteases.high molecular (above 10 kda) melanoidins isolated from coffee beans of varying roasting degree were found to be efficient inhibitors for the zinc-containing matrix metalloproteases mmp-1, mmp-2, and mmp-9 with ic(50) values ranging between 0.2 and 1.1 mg/ml in vitro. the inhibitory potential increased with roasting degree. no or only slight inhibition of other zinc-containing peptidases closely related to mmps, namely, clostridium histolyticum collagenase and angiotensin converting enzyme, was ...201121961901
effect of collagen digestion on the passive elastic properties of diaphragm muscle in rat.effects of collagen digestion have been defined up to the fibril level. however, the question remains as to whether the alteration of skeletal muscle extracellular matrix (ecm) affects a muscle's passive elastic response. various elastography methods have been applied as tools for evaluating the mechanical properties and ecm content of skeletal muscle. in an effort to develop an ecm altered skeletal muscle model, this study determined the effect of collagen digestion on the passive elastic prope ...201019945332
mechanism of the calcium-induced trans-cis isomerization of a non-prolyl peptide bond in clostridium histolyticum collagenase.cis peptide bonds in proteins are often rate-limiting steps in protein folding or conformational change and are frequently stabilized by metal ions. in the collagen-binding domain of clostridium histolyticum collagenase, the binding of calcium ions triggers the formation of a cis peptide bond. we present free energy simulations of the formation of this cis peptide bond using a combined quantum mechanics/molecular mechanics approach together with adaptive umbrella sampling. from these simulations ...201020499930
evaluation of the tissue toxicity of antiseptics by the hen's egg test on the chorioallantoic membrane (hetcam).antiseptics are frequently used for the prophylaxis and treatment of local infections of chronic wounds. whereas local antiseptics in general have a positive effect on wound healing an uncritical use may impair wound healing due to toxic side effects.201020562059
in vitro effects of synbiotic fermentation on the canine faecal microbiota.stirred, ph-controlled anaerobic batch cultures were used to investigate the in vitro effects of galacto-oligosaccharides (gos) alone or combined with the probiotic bifidobacterium bifidum 02 450b on the canine faecal microbiota of three different donors. gos supported the growth of b. bifidum 02 450b throughout the fermentation. quantitative analysis of bacterial populations by fish revealed significant increases in bifidobacterium spp. counts (bif164) and a concomitant decrease in clostridium ...201020579101
effect of a new synbiotic mixture on atopic dermatitis in infants: a randomized-controlled trial.clinical trials investigating the therapeutic effect of probiotics on atopic dermatitis (ad) show inconsistent results. better results can possibly be achieved by combining probiotics with prebiotics, i.e. synbiotics.201020184604
successful human islet isolation and transplantation indicating the importance of class 1 collagenase and collagen degradation activity assay.purified tissue dissociation enzymes (tdes) are critical to successful human islet isolation required for clinical transplantation, but little is known about the characteristics of the key enzymes-class i (c1) and class ii (c2) collagenase from clostridium histolyticum-used in these procedures. here, we show the differences between the c1 collagenase found in purified collagenase products manufactured by three suppliers and the impact of differences in c1 between two suppliers on human islet yie ...201020300051
actions of the japanese pancreas and islet transplantation association regarding transplanted human islets isolated using liberase hi.the potential for introducing transmissible spongiform encephalopathy (tse) into islet cells was indicated by recognizing that liberase hi is isolated from clostridium histolyticum grown in media containing brain-heart infusion broth. a national team within the japanese pancreas and islet transplantation association implemented an islet transplantation program in japan using liberase hi. the program comprised 65 islet isolations from non-heart-beating donors and 34 transplants into 18 patients. ...201021168667
bacterial, scfa and gas profiles of a range of food ingredients following in vitro fermentation by human colonic microbiota.it is now apparent that there is a strong link between health and nutrition and this can be seen clearly when we talk of obesity. the food industry is trying to capitalise on this by adapting high sugar/fat foods to become healthier alternatives. in confectionery food ingredients can be used for a range of purposes including sucrose replacement. many of these ingredients may also evade digestion in the upper gut and be fermented by the gut microbiota upon entering the colon. this study was desig ...201020553905
comparison of human islet isolation outcomes using a new mammalian tissue-free enzyme versus collagenase nb-1.after the discontinuation of the manufacturing liberase hi because of a small potential for prion disease transmission, roche diagnostics (indianapolis, in) developed a new enzyme product (liberase mtf [mammalian tissue free]), which is similar to liberase hi with the exception that no mammalian tissue is used in the manufacture of the collagenase component. we report our experience using the mtf enzyme in clinical islet isolations compared with serva nb-1 with modified enzyme delivery method.201020463640
new treatment options for dupuytren's surgery: collagenase and percutaneous aponeurotomy. 201020434276
new treatment option for dupuytren's contracture. 201021033070
purification and characterization of a clostripain-like protease from a recombinant clostridium perfringens culture.clostridium perfringens produces a homologue of clostripain (clo), the arginine-specific endopeptidase of clostridium histolyticum. to determine the biochemical and biological properties of the c. perfringens homologue (clp), it was purified from the culture supernatant of a recombinant c. perfringens strain by cation-exchange chromatography and ultrafiltration. analysis by sds-page, n-terminal amino acid sequencing and tof mass spectrometry revealed that clp consists of two polypeptides compris ...201019850615
intestinal dysbiosis and reduced immunoglobulin-coated bacteria associated with coeliac disease in children.coeliac disease is a chronic intestinal inflammatory disorder due to an aberrant immune response to dietary gluten proteins in genetically predisposed individuals. mucosal immune response through iga secretion constitutes a first line of defence responsible for neutralizing noxious antigens and pathogens. the aim of this study was the characterization of the relationships between immunoglobulin-coated bacteria and bacterial composition of faeces of coeliac disease (cd) patients, untreated and tr ...201020181275
distribution of polyethylene wear particles and bone fragments in periprosthetic tissue around total hip joint replacements.ultra-high molecular weight polyethylene (uhmwpe) wear particles play a significant role in failures of total joint replacements (tjrs). in this work, we investigated the distribution of these wear particles in periprosthetic tissues obtained from nine revisions of hip tjr. in the first step, all periprosthetic tissues were combined and mechanically separated into granuloma tissue (containing hard granules visible to the naked eye) and surrounding tissue (without visible granules). in the second ...201020417318
diets supplemented with chickpea or its main oligosaccharide component raffinose modify faecal microbial composition in healthy adults.the effects of diets supplemented with either chickpea or its main oligosaccharide raffinose on the composition of the faecal microbial community were examined in 12 healthy adults (18-65 years) in a randomised crossover intervention study. subjects consumed their usual diet supplemented with soups and desserts that were unfortified, or fortified with either 200 g/d of canned chickpeas or 5 g/d of raffinose for 3 week periods. changes in faecal bacterial populations of subjects were examined usi ...201021831757
intestinal microbiota in allergic and nonallergic 1-year-old very low birth weight infants after neonatal glutamine supplementation.previously, glutamine-enriched enteral nutrition in very low birth weight infants (vlbw) decreased the incidence of atopic dermatitis at age 1 year. the aim of this study was to determine whether this effect is related to changes in intestinal bacterial species that are associated with allergy, such as bifidobacteria, clostridium histolyticum, clostridium lituseburense (chis/lit group) and escherichia coli at age 1 year.201020626364
tissue dissociation enzyme neutral protease assessment.neutral proteases, essential components of purified tissue dissociation enzymes required for successful human islet isolation, show variable activities and effects of substrate on their activities. initially we used a spectrophotometric endpoint assay with azocasein substrate to measure neutral protease activity. after critical review of the results, we observed these data to be inconsistent and not correlating expected differences in specific activities between thermolysin and bacillus polymyxa ...201020692405
collagenase clostridium histolyticum for dupuytren's contracture.dupuytren's disease is a non-malignant, progressive disorder of the hands that can severely limit hand function and diminish overall quality of life. with global life expectancy increasing, the prevalence of this disease appears to be increasing amongst all ethnic groups. treatment has traditionally remained surgical with few effective, nonsurgical options. however, with the introduction of collagenase clostridium histolyticum to treat dupuytren's contractures, physicians and surgeons may be pro ...201020666587
grape seed proanthocyanidins increase collagen biodegradation resistance in the dentin/adhesive interface when included in an adhesive.contemporary methods of dentin bonding could create hybrid layers (hls) containing voids and exposed, demineralised collagen fibres. proanthocyanidins (pa) have been shown to cross-link and strengthen demineralised dentin collagen, but their effects on collagen degradation within the hl have not been widely studied. the purpose of this study was to compare the morphological differences of hls created by bisgma/hema model adhesives with and without the addition of grape seed extract pa under cond ...201020709136
mammalian tissue-free liberase: a new gmp-graded enzyme blend for human islet isolation. 201020683428
the effect of truncated collagenase class i isomers on human islet isolation outcome. 201020683429
a new method to value efficiency of enzyme blends for pancreatic tissue digestion.islet transplantation, since the 1990s, has been an example of human cell therapy. nevertheless, the islet isolation procedure is not completely standardized; in fact, >50% of islet procedures do not eventuate in transplantation due both to the variability of a donor's pancreas and to the unpredictable efficiency of an enzymatic blend. the enzymes used in pancreas isolation to digest several substrates are extracted from clostridium histolyticum. in particular, they have strong collagenolytic ac ...201020692403
high-level production and purification of clostripain expressed in a virulence-attenuated strain of clostridium perfringens.clostripain (clo) produced by clostridium histolyticum is an arginine-specific endopeptidase with the potential for applicability to diverse medical and industrial uses. in this study, we developed an expression system allowing high-level production and efficient purification of recombinant clo (rclo). our expression system comprises pclo, an rclo expressing vector, and clostridium perfringens 13δ6, an in-frame deletion strain as to six genes encoding major virulence factors and secretory protei ...201020940055
scientific understanding and clinical management of dupuytren disease.dupuytren disease (dd) is a fibroproliferative disorder of unknown etiology that often results in shortening and thickening of the palmar fascia, leading to permanent and irreversible flexion contracture of the digits. this review provides a detailed update of the scientific understanding of dd and its clinical management, with perspectives on emerging research and therapy. established risk factors include genetic predisposition and ethnicity, as well as sex and age. several environmental risk f ...201021060335
injectable collagenase clostridium histolyticum: a new nonsurgical treatment for dupuytren's disease.the collagenase option for the reduction of dupuytren's (cord) ii study investigated the efficacy and safety of injectable xiaflex (collagenase clostridium histolyticum), in patients with dupuytren's contracture.201021134613
new drugs: sipuleucel-t, cabazitaxel, and collagenase clostridium histolyticum. 201021071329
occult caecal malignancy leading to mycotic abdominal aortic aneurysm rupture. 201021114723
determination of the in vivo prebiotic potential of a maize-based whole grain breakfast cereal: a human feeding study.epidemiological studies have shown an inverse relationship between risk of cvd and intake of whole grain (wg)-rich food. regular consumption of breakfast cereals can provide not only an increase in dietary wg but also improvements to cardiovascular health. various mechanisms have been proposed, including prebiotic modulation of the colonic microbiota. in the present study, the prebiotic activity of a maize-derived wg cereal (wgm) was evaluated in a double-blind, placebo-controlled human feeding ...201020487589
the emerging role of clostridium histolyticum collagenase in the treatment of dupuytren disease.dupuytren disease (dd) is a common fibroproliferative disease of unknown etiopathogenesis affecting the palmar aponeurosis, causing reduced hand function and resulting in fixed flexion contractures of the digits. current gold standard treatment for the management of dd is surgical excision involving removal of the affected palmar fascial tissue. however, there are potential complications associated with surgery as it is costly and a positive surgical outcome is often short-lived because the dise ...201021127696
bioactive peptides derived from vascular endothelial cell extracellular matrices promote microvascular morphogenesis and wound healing in vitro.studies in our laboratory indicate that collagenase from clostridium histolyticum promotes endothelial cell and keratinocyte responses to injury in vitro and wound healing in vivo. we postulate that matrix degradation by clostridial collagenase creates bioactive fragments that can stimulate cellular responses to injury and angiogenesis. to test this hypothesis, we performed limited digestion of defined capillary-endothelial-derived extracellular matrices using purified human or bacterial collage ...201021134032
biochemical characterization of the catalytic domains of three different clostridial collagenases.clostridial collagenases are used for a broad spectrum of biotechnological applications and represent prime target candidates for both therapy and diagnosis of clostridial infections. in this study, we biochemically characterized the catalytic domains of three clostridial collagenases, collagenase g (colg) and h (colh) from clostridium histolyticum, and collagenase t (colt) from c. tetani. all protein samples showed activity against a synthetic peptidic substrate (furylacryloyl-leu-gly-pro-ala, ...200918937627
shifts in clostridia, bacteroides and immunoglobulin-coating fecal bacteria associated with weight loss in obese adolescents.to evaluate the effects of a multidisciplinary obesity treatment programme on fecal microbiota composition and immunoglobulin-coating bacteria in overweight and obese adolescents and their relationship to weight loss.200919050675
pfizer swallows wyeth, validates niche buster. 200919270661
purification and characterization of an eggshell membrane decomposing protease from pseudomonas aeruginosa strain me-4.a bacterial strain, me-4, isolated from farm soil and identified as pseudomonas aeruginosa, grew well on a medium containing eggshell membrane (esm). p. aeruginosa strain me-4 decomposed the esm by producing an extracellular protease able to solubilize it. the protease was purified to homogeneity from culture supernatant by fractionation with (nh(4))(2)so(4), as well as cm52 cellulose and de52 cellulose column chromatography, with a final yield of 47%. the molecular mass of the enzyme was 33 kda ...200919332295
a universal strategy for high-yield production of soluble and functional clostridial collagenases in e. coli.clostridial collagenases are foe and friend: on the one hand, these enzymes enable host infiltration and colonization by pathogenic clostridia, and on the other hand, they are valuable biotechnological tools due to their capacity to degrade various types of collagen and gelatine. however, the demand for high-grade preparations exceeds supply due to their pathogenic origin and the intricate purification of homogeneous isoforms. we present the establishment of an escherichia coli expression system ...200919333597
deformation-dependent enzyme mechanokinetic cleavage of type i collagen.collagen is a key structural protein in the extracellular matrix of many tissues. it provides biological tissues with tensile mechanical strength and is enzymatically cleaved by a class of matrix metalloproteinases known as collagenases. collagen enzymatic kinetics has been well characterized in solubilized, gel, and reconstituted forms. however, limited information exists on enzyme degradation of structurally intact collagen fibers and, more importantly, on the effect of mechanical deformation ...200919388774
collagenase santyl ointment: a selective agent for wound debridement.enzymatic debridement is a frequently used technique for removal of necrotic tissue from wounds. proteases with specificity to break down the collagenous materials in necrotic tissues can achieve selective debridement, digesting denatured collagen in eschar while sparing nonnecrotic tissues. this article provides information about the selectivity of a collagenase-based debriding agent, including evidence of its safe and efficacious uses. recent research has been conducted, investigating the chem ...200919918145
clinical applications for the use of enzymatic debriding ointment and broad-spectrum bacteriostatic foam dressing.the optimal management of a chronic, nonhealing wound is challenging to experienced and novice clinicians alike. treatment must be individualized, but principles of wound bed preparation, including debridement of necrotic tissue, maintenance of moisture in the wound, and prevention of bacterial overgrowth, guide the management of nonhealing wounds caused by a variety of different etiologies. this article summarizes our experiences with 2 products, collagenase ointment and crystal violet/methylen ...200919918146
exopolysaccharides produced by bifidobacterium longum ipla e44 and bifidobacterium animalis subsp. lactis ipla r1 modify the composition and metabolic activity of human faecal microbiota in ph-controlled batch cultures.exopolysaccharides (eps) isolated from two bifidobacterium strains, one of human intestinal origin (bifidobacterium longum subsp. longum ipla e44) and the other from dairy origin (bifidobacterium animalis subsp. lactis ipla r1), were subjected to in vitro chemically simulated gastrointestinal digestion, which showed the absence of degradation of both polymers in these conditions. polymers were then used as carbon sources in ph-controlled faecal batch cultures and compared with the non-prebiotic ...200919735956
inhibitive effects of alkyl gallates on hyaluronidase and collagenase.a series of the gallate esters of n-alkanols (c(1)-c(12)) was examined to determine their inhibitory activities against hyaluronidase and collagenase. hexyl, heptyl, octyl, nonyl, and decyl gallates inhibited both hyaluronidase and collagenase, and the most potent inhibitor was octyl gallate against both enzymes. octyl 3,5-dihydroxybenzoate showed inhibitory effects on hyaluronidase, whereas collagenase was inhibited by octyl 3,4-dihydroxybenzoate.200919809167
unidirectional binding of clostridial collagenase to triple helical substrates.histotoxic clostridia produce collagenases responsible for extensive tissue destruction in gas gangrene. the c-terminal collagen-binding domain (cbd) of these enzymes is the minimal segment required to bind to collagen fibril. collagen binding efficiency of cbd is more pronounced in the presence of ca(2+). we have shown that cbd can be functional to anchor growth factors in local tissue. a (1)h-(15)n hsqc nmr titration study with three different tropocollagen analogues ((pog)(10))(3), ((gpog)(7) ...200919208618
ca2+-induced linker transformation leads to a compact and rigid collagen-binding domain of clostridium histolyticum collagenase.clostridium histolyticum collagenase is responsible for extensive tissue destruction in gas gangrene, and its activity is enhanced by calcium ions. the collagen-binding domain is the minimal segment of the enzyme required for binding to insoluble collagen fibrils and for subsequent collagenolysis. the collagen-binding domain is joined to another binding module by a conserved 14-amino-acid linker. the linker undergoes secondary structural transformation from an alpha-helix to a beta-strand and fo ...200919490118
mechanical strain enhances survivability of collagen micronetworks in the presence of collagenase: implications for load-bearing matrix growth and stability.there has been great interest in understanding the methods by which collagen-based load-bearing tissue is constructed, grown and maintained in vertebrate animals. to date, the responsibility for this process has largely been placed with mesenchymal fibroblastic cells that are thought to fully control the morphology of load-bearing extracellular matrix (ecm). however, given clear limitations in the ability of fibroblastic cells to precisely place or remove single collagen molecules to sculpt tiss ...200919657003
injectable collagenase clostridium histolyticum for dupuytren's contracture.dupuytren's disease limits hand function, diminishes the quality of life, and may ultimately disable the hand. surgery followed by hand therapy is standard treatment, but it is associated with serious potential complications. injection of collagenase clostridium histolyticum, an office-based, nonsurgical option, may reduce joint contractures caused by dupuytren's disease.200919726771
the in vitro effect of a collagenolytic enzyme inhibitor on lesion development in root dentin.to evaluate the effects of a new active collagenase inhibitor, pirocton olamine (po), on acid demineralization in dentin and to investigate possible mechanisms of the inhibitory effects.200919626976
injectable collagenase clostridium histolyticum for dupuytren's contracture. 200920032329
injectable collagenase clostridium histolyticum for dupuytren's contracture. 200920050321
structure of the claudin-binding domain of clostridium perfringens enterotoxin.clostridium perfringens enterotoxin is a common cause of food-borne and antibiotic-associated diarrhea. the toxin's receptors on intestinal epithelial cells include claudin-3 and -4, members of a large family of tight junction proteins. toxin-induced cytolytic pore formation requires residues in the nh(2)-terminal half, whereas residues near the cooh terminus are required for binding to claudins. the claudin-binding cooh-terminal domain is not toxic and is currently under investigation as a pote ...200817977833
an internally quenched fluorescent substrate for collagenase.a synthetic collagenase substrate containing the internal peptide sequence--gly-gly-pro-leu-gly-pro-pro-gly-pro--has been synthesized, with an n-terminus 4-((4-(dimethylamino)phenyl)azo)-benzoyl (dabcyl) group and c-terminus 5-[2-(acetamido)ethylamino] naphthalene-1-sulfonic acid (aedans) moiety resulting in internal quenching of aedans fluorescence. peptide bond hydrolysis results in a large increase in fluorescence at 490 nm upon excitation at 336 nm. the substrate is cleaved exclusively by cl ...200818260138
development and characterization of a collagen degradation assay to assess purified collagenase used in islet isolation.a collagen degradation activity (cda) assay was developed to improve the biochemical characterization of purified collagenase used for islet isolation.200818374061
in vitro fermentation of oat and barley derived beta-glucans by human faecal microbiota.fermentation of beta-glucan fractions from barley [average molecular mass (mm), of 243, 172, and 137 kda] and oats (average mm of 230 and 150 kda) by the human faecal microbiota was investigated. fractions were supplemented to ph-controlled anaerobic batch culture fermenters inoculated with human faecal samples from three donors, in triplicate, for each substrate. microbiota changes were monitored by fluorescent in situ hybridization; groups enumerated were: bifidobacterium genus, bacteroides an ...200818430007
crystallization and preliminary x-ray characterization of the catalytic domain of collagenase g from clostridium histolyticum.the catalytic domain of collagenase g from clostridium histolyticum has been cloned, recombinantly expressed in escherichia coli and purified using affinity and size-exclusion column-chromatographic methods. crystals of the catalytic domain were obtained from 0.12 m sodium citrate and 23%(v/v) peg 3350 at 293 k. the crystals diffracted to 2.75 a resolution using synchrotron radiation. the crystals belong to an orthorhombic space group, with unit-cell parameters a = 57, b = 109, c = 181 a. this u ...200818453715
1h, 13c and 15n resonance assignments of ca2+ bound collagen-binding domain derived from a clostridial collagenase.(1)h, (15)n, and (13)c nmr assignments for 116 amino acids (gly893-lys1008) of a bacterial collagen-binding domain (cbd) derived from clostridium histolyticum class i collagenase were accomplished. clostridial collagenases hydrolyze insoluble collagen. one to three copies of collagen-binding domains (cbds) are present at their c-termini, each of which is the minimal segment required for the binding to the insoluble substrate. cbd has been shown to be able to anchor fused growth factors for up to ...200819636886
broad-range pcr as a supplement to culture for detection of bacterial pathogens in patients with a clinically diagnosed spinal infection.we aimed to evaluate broad-range pcr and subsequent sequencing compared to conventional culture in the diagnosis of spinal infection. the method was a prospective study of all patients admitted to aarhus university hospital for surgery during a 12-months period with a clinically diagnosed infection of the spine. samples from patients undergoing surgery for non-infectious causes (malignancy etc.) were included as control group. specimens were submitted to conventional culture and molecular invest ...200818609207
high-level expression of his-tagged clostridial collagenase in clostridium perfringens.clostridium histolyticum collagenase is used to isolate cells from various organs and tissues for tissue engineering, and also to treat destructive fibrosis; thus, the demand for high-grade enzyme preparations is increasing. in this study, we constructed a plasmid encoding c. histolyticum type ii collagenase (colh) with a c-terminal hexahistidine tag (colh-his) to facilitate the purification of the enzyme through immobilized metal affinity chromatography (imac). when colh-his was expressed in a ...200818629492
[molecular diagnosis of osteogenesis imperfecta type i].osteogenesis imperfecta (oi) is caused by mutations in collagen type i genes. in contrast to oi type ii, iii and iv where there are the structural mutations, in oi type i decreased production of normal collagen is due to the presence of a null allele. because both pharmacological and gene therapy approaches depend on type of mutation and its consequences, quick and proper diagnosis is required.200819145934
[a mass-spectrometric approach to primary screening of collagenolytic enzymes].a comparative analysis of maldi tof mass spectra of low-molecular products resulting from the hydrolysis of native collagen i by collagenases of various classes (bacterial metallocollagenase from clostridium histolyticum, serine collagenase from the morikrasa commercial preparation, cysteine collagenase from serratia proteomaculans, and cysteine collagenases from larvae of beetles dermestesfrischi and d. maculates) was carried out. the spectra contain a number of ion peaks common for all collage ...200818672690
experimental model of warfarin-associated intracerebral hemorrhage.future demographic changes predict an increase in the number of patients with atrial fibrillation. as long-term anticoagulation for the prevention of ischemic strokes becomes more prevalent, the burden of warfarin-associated intracerebral hemorrhage (w-ich) is likely to grow. however, little is known about the clinical aspects and pathophysiologic mechanisms of w-ich. this study describes the development of a mouse model of w-ich in which hematoma growth and outcomes can be correlated with antic ...200818772448
a method of isolating the collagen (i) alpha2 chain carboxytelopeptide for species identification in bone fragments.we present a novel method for the isolation and analysis of the bone collagen (i) alpha2 chain carboxytelopeptide as a species biomarker. conventional methods for the analysis and sequencing of mixtures of proteins and peptides commonly involve using the protease trypsin to cleave proteins present in the sample. however, in the study of collagen, these methods result in very complex mixtures of peptides that are difficult to analyze and the acquired results are not reproducible. here we use bact ...200818164255
molecular analysis of the digestive microbiota in a gnotobiotic mouse model during antibiotic treatment: influence of saccharomyces boulardii.the probiotic saccharomyces boulardii is a non-pathogenic yeast that has been proven efficient in the prevention of antimicrobial-associated diarrhea and of clostridium difficile associated colitis. we evaluated the influence of the administration of s. boulardii on the composition of the fecal microbiota in a human microbiota-associated mouse model. this evaluation was run before, during and after a 7-day oral treatment with amoxicillin clavulanic acid. predominant groups of bacteria were quant ...200818511310
detrimental effect of excessive collagenase class ii on human islet isolation outcome.commercially available purified collagenases, derived from clostridium histolyticum, contain two different classes of collagenase: class i collagenase (ci) and class ii collagenase (cii) at a predetermined ratio. in this study, using purified ci and cii in separate vials, we had a unique opportunity to investigate the effect of the proportion between two collagenase classes on clinical human islet isolation. pancreas organs derived from deceased donors were prospectively assigned to one of three ...200818680482
lethal factor of clostridium histolyticum kills cells by apoptosis.in this study, for the first time, clostridium histolyticum lethal factor was purified by ammonium sulfate precipitation of culture broth, centrifugation through an amicon filter device and hydrophobic interaction chromatography. the purified lethal factor was devoid of proteolytic activity. at a concentration of 150 ng ml(-1) the lethal factor killed 50% of hela cells within 24 h of exposure. abrogation of actin filaments, activation of caspases, fragmentation of nuclear dna as well as ultrastr ...200717328764
characterization of the gastrointestinal mucosa-associated microbiota of pigs and chickens using culture-based and molecular methodologies.the microbiota of the gastrointestinal tract (git) can play an important role in the healthy status of farm animals and in the safety of the whole food chain. in this study, the mucosa-associated microbiota of the git of pigs and chickens was analyzed by culture methods and fluorescence in situ hybridization combined with flow cytometry (fcm-fish). in all pig git sections, lactic acid bacteria, enterobacteriaceae, atopobium, bacteroides, and clostridium histolyticum were the predominant bacteria ...200718095433
analysis of contribution of collagen fibre component in viscoelastic behaviour of periodontal ligament using enzyme probe.the aims of this study are to observe microscopic changes in the periodontal ligament (pdl) collagen fibres after collagenase treatment, to analyse stress-relaxation behaviour of pdl specimens treated with collagenase, and to elucidate the contribution of the collagen component to the viscoelastic behaviour of the pdl. transverse sections of rat mandibular first molars (n=24) were treated in vitro with 0, 8, 16, or 24 units of bacterial collagenase for 4h at 37 degrees c. histological specimens ...200717266965
factors influencing the collagenase digestion phase of human islet isolation.substantial advances in human islet isolation technology have occurred during the past decade. however, it is still difficult to recover the entire quantity of islets contained in a pancreas. a major obstacle to successful human islet isolation has been the variability of the collagenase digestion phase of islet isolation. future advances in enzyme technology will make it possible to optimally liberate islets with enzyme blends "tailor-made" for each individual donor pancreas. such innovative st ...200717220782
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