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Limulus amebocyte lysate (LAL) is an aqueous extract of blood cells (amoebocytes) from the Atlantic horseshoe crab, Limulus polyphemus. LAL reacts with bacterial endotoxin lipopolysaccharide (LPS), which is a membrane component of gram-negative bacteria. This reaction is the basis of the LAL test, which is then used for the detection and quantification of bacterial endotoxins. Fred Bang reported in 1956 that gram-negative bacteria, even if killed, will cause the blood of the horseshoe crab to turn into a semi-solid mass. It was later recognized that the animals blood cells, mobile cells called amoebocytes, contain granules with a clotting factor known as coagulogen; this is released outside the cell when bacterial endotoxin is encountered. The resulting coagulation is thought to contain bacterial infections in the animals semi-closed circulatory system. In 1970 the U.S. Food and Drug Administration (FDA) approved LAL for testing drugs, products and devices that come in contact with the blood. ...

PubMed Central Canada (PMC Canada) provides free access to a stable and permanent online digital archive of full-text, peer-reviewed health and life sciences research publications. It builds on PubMed Central (PMC), the U.S. National Institutes of Health (NIH) free digital archive of biomedical and life sciences journal literature and is a member of the broader PMC International (PMCI) network of e-repositories.

If you have ever taken medication, received a vaccine, or had a surgical implant, you should thank a horseshoe crab. These prehistoric-looking animals are actually really important to modern medicine. But why?. Its all about their blue blood. Mammals have hemoglobin in their blood, which contains iron- hence the red color. But horseshoe crabs transport oxygen through their bodies via hemocyanin, which contains copper, making their blood blue.. Even more interesting is a compound in the crabs blood called Limulus Amebocyte Lysate, or LAL. LAL binds to bacteria, viruses and fungi and acts to protect the animals system from infection. Its worked pretty well- horseshoe crabs have been around since about 100 million years BEFORE the dinosaurs, and theyre still going strong!. This ability to bind endotoxins makes horseshoe crab blood incredibly useful- and valuable. LAL is the worldwide standard screening test for bacterial contamination, and its used to test drugs, vaccines and surgical ...

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Lot specific results are available in the Certificate of Analysis, which is available at www.atcc.org or by contacting ATCC technical service. SPECIFICATION CERTIFICATE Test Specification pH 1 7.1 to 7.4 Osmolality 2 256 to 286 mOsm/kg Endotoxin 3 ≤ 0.25 EU/mL Mycoplasma 4 Tested Negative Sterility 5 Pass *Please consult the Certificate of Analysis for lot-specific test results. Measured with a standard pH meter.  Meters are calibrated daily using standards traceable to the National Institute of Standards Technology using guidelines outlined in the current edition of USP. Measured with a standardized osmometer, i.e. freezing point depression.   Meters are calibrated daily using standards traceable to the National Institute of Standards Technology using guidelines outlined in the current edition of USP. Chromogenic LAL Assay using guidelines outlined by the manufacturer & FDA (December 1987), “Validation of the Limulus Amebocyte Lysate Test as an End Product Endotoxin Test for

Lot specific results are available in the Certificate of Analysis, which is available at www.atcc.org or by contacting ATCC technical service. SPECIFICATION CERTIFICATE Test Specification pH 1 7.1 to 7.4 Osmolality 2 256 to 286 mOsm/kg Endotoxin 3 ≤ 0.25 EU/mL Mycoplasma 4 Tested Negative Sterility 5 Pass *Please consult the Certificate of Analysis for lot-specific test results. Measured with a standard pH meter.  Meters are calibrated daily using standards traceable to the National Institute of Standards Technology using guidelines outlined in the current edition of USP. Measured with a standardized osmometer, i.e. freezing point depression.   Meters are calibrated daily using standards traceable to the National Institute of Standards Technology using guidelines outlined in the current edition of USP. Chromogenic LAL Assay using guidelines outlined by the manufacturer & FDA (December 1987), “Validation of the Limulus Amebocyte Lysate Test as an End Product Endotoxin Test for

Objective:To determine the bacterial edotoxins in danshen injection.Methods:After interfering test.Cel-dot method of tachypleus amebocyte(TAL) test was applied to detect bacterial endotoxins in danshen injection.Results:The sample 60-fold diluents from three batchs of danshen injection did not interfere the test for bacterial endotoxins.The content of bacterial endotoxins in all test samples was not more than 15 EU·ml-1.Conclusion:TAL test provides a new way to detect bacterial endotoxins in danshen injection.

Cell culture. HT-29 cells obtained from the American Type Culture Collection (Rockville, MD) were maintained in McCoys medium with l-glutamine (Mediatech, Washington, DC), supplemented with 10% FCS. Cells were kept at subconfluence in a humidified incubator at 37°C with 5% CO2.. Reagents and antibodies. The monoclonal antibody (Ab) to Fas, CH-11, was purchased from Kamiya Biomedical (Thousand Oaks, CA), and subsequently CH-11 was purchased from Upstate Biotechnology (Lake Placid, NY). The level of endotoxin contamination for CH-11 is 0.048 EU/μg as determined by the limulus amebocyte lysate test. IFN-γ was purchased from R&D Systems (Minneapolis, MN). For Fas surface staining, biotin-conjugated anti-human CD95 (Fas/Apo-1), biotin-conjugated anti-human Fas ligand, and streptavidin-phycoerythrin (PE) were purchased from Pharmingen. The ApoBrDU apoptosis detection kit was purchased from Pharmingen and was used as per the manufacturers directions. For Jun, Fos, and nuclear factor-κB (NF-κB) ...

Lipopolysaccharide (LPS) extracted from eight strains of Yersinia pestis, which had been cultured at 28 or 37 degreesC, reacted equally well, in Western blots, with four monoclonal antibodies generated against the LPS from a single strain of Y. pestis cultured at 28 degreesC. LPS was extracted and purified from Y. pestis strain GB, which had been cultured at 28 degreesC. When the LPS was analysed by SDS-PAGE and MALDI-TOF mass spectrometry it was found to be devoid of an O-antigen. The LPS possessed activity of 2.7 endotoxin units/ng in the Limulus amoebocyte lysate assay. The LPS stimulated the production of TNF alpha and IL-6 from mouse macrophages, but was less active in these assays than LPS isolated from Escherichia coli strain 0111. Y. pestis LPS, either alone or with cholera toxin B subunit, was used to immunize mice. Either immunization schedule resulted in the development of an antibody response to LPS. However, this response did not provide protection against 100 MLD of Y. pestis ...

Corning Matrigel matrix is a solubilized basement membrane preparation extracted from the Engelbreth-Holm-Swarm (EHS) mouse sarcoma, a tumor rich in extracellular matrix proteins, including Laminin (a major component), Collagen IV, heparan sulfate proteoglycans, entactin/nidogen, and a number of growth factors. Quality Mouse colonies routinely screened for pathogens via Mouse Antibody Production (MAP) testing Extensive PCR testing is performed to screen for a number of pathogensTested and found negative for bacteria, fungi, and mycoplasma Protein concentrations determined by Lowry method Endotoxin units measured by Limulus Amoebocyte Lysate assay Matrigel Matrix gel stability tested for a period of 14 days at 37°C Biological activity determined for each lot using a neurite outgrowth assay. Source Engelbreth-Holm-Swarm Mouse Tumor Preparation and Storage Store at -20°C. Avoid multiple freeze-thaws. Application Cell Growth and Differentiation Metabolism/Toxicology Studies Invasion Assays in vitro and in

Kevin Williams from bioMerieux discusses the recombinant Factor C assay that has been developed to help alleviate the pressures of a growing demand for the Limulus amoebocyte lysate assay

This proposal will use the resources of Tachypleus tridentatus (horseshoe crab) to develop the reagent for Limulus test, adjuvant for animal vaccines, cancer-cell inhibitor, bacteria growth inhibitor in a three-year study. The hemolymph from horseshoe crab contains a major component of lipopolysaccharide (LPS)-binding protein and which has been characterized and developed for the Limulus test to detect the bacterial contaminant, a cell-wall component of gram negative bacteria, in medical reagent used for humans. The sensitivity of Limulus test was compared to the traditional test by injecting the specimens into rabbits with higher sensitivity of 250 fold. Previously, the Penchu Marine Biology Research Center, council of Agriculture, has investigated the natural resource of horseshoe crab in the ocean of Penchu region and studied the performance of horseshoe crab including the breeding, feeding and collection of hemolymph. Based on their studies, we will use the hemolymph components to develop ...

Structural changes underlying exocytosis evoked by the application of endotoxin to Limulus amebocytes were studied at the level of detail afforded by freeze-fracture and freeze-substitution techniques combined with the time resolution of direct rapid-freezing. The results with amebocytes prepared in this manner differed from those with other secretory cells prepared by conventional means. Exocytosis begins within seconds of endotoxin treatment when the plasmalemma invaginates to form pedestallike appositions with peripheral secretory granules. The juxtaposed membranes at these pedestal appositions form several punctate pentalaminar contacts, but examination of freeze-fractured pedestals failed to reveal any corresponding changes in the intramembrane particle distribution. Small secretory granule openings or pores, which are very infrequent, appear within the first 5 s after endotoxin treatment. These pores rapidly widen and this widening is immediately followed by the sequential dissolution of the

The measurement of airborne endotoxins is thus far not standardized. Earlier studies reported higher endotoxin yields when Tween 20 was added to the media used for filter extraction and in the Limulus amebocyte lysate (LAL) assay. This study compared four common media and assessed the effects of Tween during extraction and analysis separately. Parallel airborne dust samples from five work environments (n = 250) were used to compare the four media (pyrogen-free water [PFW], PFW-Tween 20, PFW-Tris, and PFW-triethyl-amine- phosphate [TAP]) and an extraction time of 10 or 60 min. A subset of the extracts in PFW or PFW-Tween (n = 40) were analyzed in parallel LAL assays with PFW or PFW-Tween as the assay medium. The results produced by a shorter extraction time or the presence of Tris were similar to the results for the reference procedure (PFW and 60 min of shaking). The use of PFW-TAP showed overall lower yields and a deviant calibration curve. The presence of Tween in the extraction medium ...

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Pyrogens are substances (such as Gram-negative and Gram-positive bacteria, fungi, and viruses) that can produce fever. Pharmaceutical products (such as fluids for injection, medical devices, and human biological products) intended for implantation or parenteral administration must be properly and accurately evaluated for the presence of pyrogenic substances and shown to be free of contamination prior to their clinical or veterinary use.. The U.S., European, and Japanese Pharmacopoeias currently recognize two test methods for pyrogen testing. The rabbit pyrogen test (USP28[151]) involves measuring the rise in temperature of rabbits following intravenous injection of a test solution. The bacterial endotoxin test (USP28[85]) is an in vitro assay based on the coagulation of Limulus amoebocyte lysate following exposure to endotoxin, a pyrogenic substance that is produced by certain bacteria. An important distinction between these two tests is that the bacterial endotoxin test detects only endotoxin ...

Gastrointestinal (GI) ischemia during exercise is associated with luminal permeability and increased systemic lipopolysaccharides (LPS). This study aimed to assess the impact of a multistrain pro/prebiotic/antioxidant intervention on endotoxin unit levels and GI permeability in recreational athletes. Thirty healthy participants (25 males, 5 females) were randomly assigned either a multistrain pro/prebiotic/antioxidant (LAB⁴ANTI; 30 billion CFU day-1 containing 10 billion CFU day-1Lactobacillus acidophilus CUL-60 (NCIMB 30157), 10 billion CFU day-1Lactobacillus acidophillus CUL-21 (NCIMB 30156), 9.5 billion CFU day-1Bifidobacterium bifidum CUL-20 (NCIMB 30172) and 0.5 billion CFU day-1Bifidobacterium animalis subspecies lactis CUL-34 (NCIMB 30153)/55.8 mg day-1fructooligosaccharides/ 400 mg day-1 α-lipoic acid, 600 mg day-1N-acetyl-carnitine); matched pro/prebiotic (LAB⁴) or placebo (PL) for 12 weeks preceding a long-distance triathlon. Plasma endotoxin units (via Limulus amebocyte lysate ...

Deteksi lan eliminasi endotoksin wis dadi masalah bertahun-taun kanggone indhustri farmasi lan piranti-piranti kadhokteran.[1] Contohe pemberian obat kang terkontaminasi;; denig endotoksin bisa nyebapake komplikasi utawa kematian marang pasien.[1] Selain itu, endotoksin uga dadi masalah kanggone wong kang makarya karo kultur sèl lan rekayasa genetika.[1] Mangka iku, dikembangake métodhe kanggo ndeteksi lan mengeliminasi endotoksin.[1] Prosedur mau kudu sensitif banget saéngga bisa ndeteksi endotoksin nganti 0,25 endotoxin unit (E.U.) utawa setara karo 0,025ng/ml.[1] Salah siji prosedur kang paling akurat ya iku uji Limulus amoebocyte lysate (LAL) kang andhedhasar marang observasi pembentukan gel beku sewaktu endotoksin bersentuhan karo protéin pembeku saka amoebocytes Limulus kang bersikulasi.[1] Perangkat uji iki kasusub saka kalsium, enzim propembekuan (proclotting) lan senyawa propenggumpalan/prokoagulan (procoagulan).[1] Enzim proclotting bakal teraktivasi déning endoktoksin lan kasium ...

Bacterial Endotoxins Test - (BET) Useful facts about endotoxins and their detection with the Limulus Amoebocyte Lysate (LAL) assays Endotoxin (LipoPolySaccharide, LPS) is produced exclusively by Gram-negative bacteria, such as Escherichia coli, Salmonella enteritidis, Legionella pneumophila, Campylobacter jejuni, Vibrio cholerae, Shigella dysenteriae, Pseudomonas aeruginosa and many, many more...

TY - JOUR. T1 - Evaluation of the activity of endotoxin trapped by a hollow-fiber dialysis membrane. AU - Yamamoto, Ken Ichiro. AU - Matsuda, Masato. AU - Hayama, Masayo. AU - Asutagawa, Jun. AU - Tanaka, Shigenori. AU - Kohori, Fukashi. AU - Sakai, Kiyotaka. PY - 2006/3/15. Y1 - 2006/3/15. N2 - All available dialysis membranes prevent endotoxin (Et) from mixing with the blood under clinical conditions. However, maintenance dialysis patients are at risk of amyloidosis attributed to Et. This suggests that Et may affect the blood even if it does not mix with the blood. The objective of the present study is to evaluate the activity of Et trapped by membranes. We made mini modules out of hollow fibers using three different types of membranes and filtered Et solution. The lumen of the hollow fibers was then filled with limulus amebocyte lysate (LAL) for 15 min at 310 K. Et activity was then determined by measuring absorbance of the LAL reagent. The surfaces of test membranes were studied using an ...

This Pyrotell®-T kinetic turbidimetric training video will help you achieve testing proficiency with Associates of Cape Cods Pyrotell®-T Limulus Amebocyte Lysate (known as LAL) reagent in the kinetic

RHMGB1 extracted from Human Embryonic Kidney 293 cells was prepared from Novoprotein. The articles of endotoxion was examined by the Novoprotein Corporation and observed to get significantly less than 0. 1 ng ug. This end result was also confirmed by our endotoxin Limulus amebocyte lysate check. Western Inhibitors,Modulators,Libraries blot evaluation was made to exclude Histone 3 protein con tamination. 50 ug rHMGB1 was diluted to 1,500 ul with saline and sterilized by filtration via a 0. 22 um sterile filter in situation of bacterial contamin ation. The dose of rHMGB1 was determined according to Qius analysis and adjusted the complete volume of injection for being 150 ul. Rats inside the management group were injected with 150 ul saline. Tissue was prepared for western blot and immunofluorescent examination.. Perfusion fixation and tissue preparation Animals had been sacrificed in accordance towards the time points of various CHIR-99021 inhibitor groups. In our pilot examine, we discovered that ...

2010. Schromm AB, Reiling N, Howe J, Wiesmuller K, Roessle M, Brandenburg K. Influence of serum on the immune recognition of a synthetic lipopeptide mimetic of the 19-kDa lipoprotein from Mycobacterium tuberculosis. INNATE IMMUNITY 2010 Aug;16(4):213-25.. Andrä J, Gutsmann T, Müller M, Schromm AB. Interactions between Lipid A and Serum Proteins. ADVANCES IN EXPERIMENTAL MEDICINE 2010 Jul;667:39-51.. Brandenburg K, Schromm AB, Gutsmann T. Endotoxins: relationship between structure, function, and activity. SUBCELLULAR BIOCHEMISTRY 2010 Jun;53:53-67.. Gutsmann T, Howe J, Zahringer U, Garidel P, Schromm AB, Koch MHJ, Fujimoto Y, Fukase K, Moriyon I, Martinez-de-Tejada G, Brandenburg K. Structural prerequisites for endotoxic activity in the Limulus test as compared to cytokine production in mononuclear cells. INNATE IMMUNITY 2010 Feb;16(1):39-47 ...

This article explains about LAL test method /procedure used to test bacterial contamination of pharmaceutical products especially parenteral preparations. Contents Aim Principle

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Materials. All chemicals were purchased from Sigma-Aldrich (St. Louis, MO) unless otherwise noted. LPS from Escherichia coli 055:B5 (lot 075K4038) with an activity of 3.3 × 106 endotoxin units/mg was used for these studies. Endotoxin activity was measured using the Limulus amebocyte assay (Cambrex, East Rutherford, NJ). PGN and LTA from Staphylococcus aureus were used for all studies. Rabbit anti-murine CD18 antiserum designed against amino acids 89 to 100 was purchased from New England Peptide (Gardner, MA). TVX was synthesized by Cayman Chemical (Ann Arbor, MI). Infinity alanine aminotransferase (ALT) reagent used to measure ALT activity was purchased from Thermo Electron Corp. (Louisville, CO).. Animals. Male C57BL/6J mice purchased from The Jackson Laboratory (Bar Harbor, ME) were used at an age of 9 to 11 weeks and weighed between 21 and 26 g. Wild-type and neutrophil elastase [NE(-/-)] knockout (B6.129X1-Ela2/J) mice were purchased from The Jackson Laboratory, used at an age of 12 to 14 ...

1. To address the question of whether endotoxaemia could be involved in the inflammatory response induced by long-term strenuous exercise, 18 male marathon runners [mean age 41±2 (SEM) years] were studied. Their performance in the marathon ranged from 2 h 46 min to 4 h 42 min. 2. Four venous blood samples were drawn: at rest, just before the race (baseline); within 15 min following the completion of the marathon; after 1 h of recovery; and the morning after the race. 3. The following humoral markers of the inflammatory response to exercise were measured: polymorphonuclear myeloperoxidase (MPO), anaphylatoxin C5a (C5a), tumour necrosis factor-α (TNF-α) and interleukin-6 (IL-6). Plasma endotoxin was measured by a sensitive and rapid chromogenic Limulus assay. All inflammatory markers were significantly increased ( P , 0.001) after the race, reaching in most cases peak values in the first blood sample drawn following the completion of the marathon [MPO, 298±19 ng/ml (SEM); C5a, 1.45±0.32 ...

This doesnt mean that microbiologists can ignore the performance of their instruments. This article sets out some of the performance characteristics which could be considered for automated LAL instruments. I am not suggesting that these all need to be looked at or that already busy microbiology lab have to grapple with yet another validation task. However, the microbiologist responsible for such systems should at least understand their strengths and limitations. Types of automated LAL instruments Automated LAL instruments fall into two broad categories depending upon the nature of the test: turbidimetric (where the rate of turbidity (or the gelation) of the lysate is proportional to the amount of endotoxin present) and chromogenic (where a synthetic yellow chromogenic substrate is used in the clotting cascade and the intensity of this yellow is related to the amount of endotoxin). Turbidimetric tests are read using photometric instruments (such as spectrophotometers) and chromogenic tests ...

There is a worldwide rise in antibiotics resistant bacteria. In the case of bacterial infection, there is therefore the need to quickly determine for each patient the susceptibility or not to antibiotics of the infecting bacteria in order to prescribe as quickly as possible the correct drug. Our patented technology based on microlevers permit in a short time span (10-30 minutes) to test ...

The coronavirus has brought significant and speedy changes to how health care services are delivered. That has further highlighted the need for rapid evaluations, with evaluators under pressure to produce timely findings to support decisions. Ahead of this weeks rapid evaluation in health care conference, Nadia Crellin and John Appleby run through some of the key issues.

After compiling a set of mutants which showed a relative increase in activity we proceeded to purify our mutant proteins. This step is crucial because it allows us to determine how our mutant compares with the wild-type on a quantitative level. For instance, if the whole cell lysate assay showed one mutant to have ten times the activity level as wild-type kumamolisin we cannot assume that there has been an increase in activity because there could simply be ten times the amount of protein with the same level of activity. To purify our mutants, we first grew them in TB and kanamycin with a single colony of the mutant. This inoculation was grown over 24 hours at 37 degrees celcius and then expanded to 50 mililiters (TB+kanamycin). We then allowed the culture to grow to a specific optical density of cells before inducing the culture using IPTG. We then allowed the cells to grow again to express kumamolisin. We then lysed the cells using a lysis buffer and centrifugation. This allowed the enzymes to ...

ENDOSAFE PTS PDF - The Endosafe®-PTS™ is a handheld spectrophotometer that utilizes FDA- licensed disposable cartridges for accurate, convenient endotoxin testing. With

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Lactated Ringers Injection USP is sterile, nonpyrogenic and contains no bacteriostatic or antimicrobial agents. This product is intended for intravenous administration in a single dose container. This solution is indicated for use in adults and pediatric patients as a source of electrolytes and water for hydration.. Not manufactured with latex, PVC or DEHP.. ***DUE TO A NATIONWIDE MANUFACTURING SHORTAGE we are unable to confirm a valid delivery date. We are currently taking orders for our Pre-Paid / Hold Waitlist. These orders will be fulfilled as the fluids become available. Please contact customer service for more information***. ...

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2000年代初期からサイケフォークやエセ密林系音楽を中心にリリースしてきたフィンランドの老舗レーベルLal Lal Lal諸作を大量に本邦初入荷しました。 本作は、ベルギーのアンビエント作家Accouが70本限定でリリースしたカセットです。アルペジオ奏法によるリズミカルなシンセアンビエント3曲を収録。既に廃盤です。 Edition of 70. Lal Lal Lal: Three beautiful synthetic trips by a Frenchman who knows the best order to press his buttons. Recorded live in Saarbrücken, Bruxelle

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Preoperative oral treatment with lactulose is used to prevent complications after surgery in patients with obstructive jaundice. The effect is perhaps the result of an inactivation of gut derived endotoxins but the exact mechanism of action is, however, unknown. Tumour necrosis factor is an important mediator of endotoxin toxicity. The cytokine tumour necrosis factor is mainly produced by mononuclear phagocytes. In this study, the effect of lactulose on the endotoxin induced tumour necrosis factor release by monocytes was investigated. The direct effect of lactulose on endotoxin was tested in a chromogenic limulus amoebocyte lysate assay. Polymyxin B a known inactivator of endotoxin was used as control in both experiments. Lactulose has a limited capacity to inactivate endotoxin as measured in the endotoxin assay. In contrast lactulose significantly reduced endotoxin induced tumour necrosis factor production by monocytes. In conclusion lactulose inhibits tumour necrosis factor production by a ...

ITG SUBJECT: BACTERIAL ENDOTOXINS/PYROGENS. Introduction. There has been considerable discussion in the literature recently pertaining to the Bacterial Endotoxins Test, its significance and interpretation, and its comparison to the USP rabbit test. The topic was previously briefly addressed via Inspection Technical Guide No. 32, dated 1/12/79, with the subject: Pyrogens, Still a Danger.. Because of the increased acceptance and use of the Bacterial Endotoxins Test, ITG No. 32 is being updated.. The USP now recognizes two tests - The Pyrogen Test conducted with rabbits and the Bacterial Endotoxins Test, also termed the Limulus Amebocyte Lysate (LAL) Test. Additionally, the agency has approved the use of the Bacterial Endotoxins Test for many drug and device products. This ITG will focus on the significance and interpretation of pyrogen/endotoxin testing. Also sources and methods of depyrogenation will be discussed. The limitations of the rabbit pyrogen test should be recognized when reviewing ...

Background Cardiovascular cell therapy represents a encouraging field, with many approaches currently being analyzed. was examined using an computerized microbial recognition program, endotoxin by a kinetic chromogenic Limulus amebocyte lysate check. T-test was utilized for record evaluation. Outcomes A fresh production technique was arranged up, centered on lean centrifugation on low denseness Ficoll-Paque, adopted by 2 cleaning actions, of which the second one at low velocity. It led to considerably higher removal of poison granulocytes and platelets, enhancing item chastity; the frequencies of Compact disc34+ cells, Compact disc133+ cells and practical hematopoietic and mesenchymal precursors had been considerably improved. The procedure was effectively authenticated relating to Great Manufacturing Methods. The producing ATMP primarily comprised of practical MNC including Compact disc34+ and Compact disc133+ cell subsets (2.98%??1.90% and 0.83%??1.32%, respectively), Compact disc184/CXCR4+ ...

BioReliance offers the rabbit pyrogen test, in addition to the LAL test, as an alternative assay for the detection of endotoxin and other pyrogens. The rabbit pyrogen test requires the injection of a small amount of batched test material into a rabbits blood stream, and monitoring for temperature increases ...

The indoor environment in animal husbandry is inevitably contaminated with endotoxin. Endotoxin exposure associates with various inflammatory illnesses in animals. The present cross-sectional study evaluated the relationship between the degree of endotoxin exposure and the cellular and humoral immune profiles of fattening pigs. Blood samples were taken from the jugular vein of 47 pigs from ten different pig farms in Korea. Whole blood cell counts and plasma immunoglobulin classes were measured. The peripheral blood mononuclear cells were stimulated in vitro with Concanavalin A for 48 hours and the cytokines released into the culture supernatants were measured. The barns in which the pigs lived were assessed for endotoxin levels in the total or respirable dust by using the Limulus Amebocyte Lysate Kinetic QCL method. Low and high endotoxin exposure were defined as ≤30 and ,30 EU/m3, respectively. Compared to pigs with low endotoxin exposure (n=19), highly exposed pigs (n=28) had higher ...

Horeshoe crabs Limulus polyphemus are remarkable living fossils which have unique blood cells (amebocytes) that are used to test human vaccines for bacterial contamination. In the 1950s, scientists at the Marine Biological Laboratory in Woods Hole, Massachusetts, not only discovered amebocytes but also found that they had special properties. If the amebocytes came into contact with bacteria, they would instantly coagulate around the bacteria and attack it. The Woods Hole scientists took this unique property of horseshoe crabs and developed a test for bacterial contamination using a horseshoe crab blood derivative called Limulus Amebocyte Lysate (LAL). This article describes discusses the medical, economic, and ecological importance of the horseshoe crab. Project Limulus is a research and community project which is working toward protecting the population of the horshoe crab.

Horseshoe crabs use hemocyanin to carry oxygen through their blood. Because of the copper present in hemocyanin, their blood is blue. Their blood contains amebocytes, which play a similar role to the white blood cells of vertebrates in defending the organism against pathogens. Amebocytes from the blood of L. polyphemus are used to make Limulus amebocyte lysate (LAL), which is used for the detection of bacterial endotoxins in medical applications.[28] This means there is a high demand for the blood, the harvest of which involves collecting and bleeding the animals, and then releasing them back into the sea. Most of the animals survive the process; mortality is correlated with both the amount of blood extracted from an individual animal, and the stress experienced during handling and transportation.[29] Estimates of mortality rates following blood harvesting vary from 3-15%[30] to 10-30%.[31][32][33] Approximately 500,000 crabs are harvested annually.[34] Bleeding may also prevent female horseshoe ...

The blood of the horseshoe crab provides a valuable medical product critical to maintaining the safety of many drugs and devices used in medical care. A protein in the blood called Limulus Amebocyte Lysate (LAL) is used by pharmaceutical and medical device manufacturers to test their products for the presence of endotoxins, bacterial substances that can cause fevers and even be fatal to humans.. The LAL test is one of the most important medical ...

Abate, W., Sattar, A. A., Liu, J., Conway, M. E. and Jackson, S. K. (2017) Evaluation of recombinant Factor C (rFC) assay for the detection of divergent LPS structural species and comparison with Limulus amebocyte lysate (LAL)-based assays and a human monocyte activity assay. Journal of Medical Microbiology, 66 (7). pp. 888-897. ISSN 0022-2615 Available from: http://eprints.uwe.ac.uk/32404 Liu, J., Abate, W., Xu, J., Corry, D., Kaul, B. and Jackson, S. K. (2011) Three-dimensional spheroid cultures of A549 and HepG2 cells exhibit different lipopolysaccharide (LPS) receptor expression and LPS-induced cytokine response compared with monolayer cultures. Innate Immunity, 17 (3). pp. 245-255. ISSN 1753-4259 Available from: http://eprints.uwe.ac.uk/12151 Liu, J., Pankhurst, L., Deacon, L., Abate, W., Hayes, E. T., Drew, G. H., Longhurst, P., Pollard, S., Longhurst, J., Tyrrel, S. and Jackson, S. K. (2011) Evaluation of inflammatory effects of airborne endotoxin emitted from composting sources. ...

INTRODUCTION: The role of microbial translocation (MT) in HIV patients living with HIV from low- and middle-income countries (LMICs) is not fully known. The aim of this study is to investigate and compare the patterns of MT in patients from Vietnam, Ethiopia and Sweden.. METHODS: Cross-sectional samples were obtained from treatment-naïve patients living with HIV-1 and healthy controls from Vietnam (n=83; n=46), Ethiopia (n=9492; n=50) and Sweden (n=51; n=19). Longitudinal samples were obtained from a subset of the Vietnamese (n=24) in whom antiretroviral therapy (ART) and tuberculostatics were given. Plasma lipopolysaccharide (LPS), sCD14 and anti-flagellin IgG were determined by the endpoint chromogenic Limulus Amebocyte Assay and enzyme-linked immunosorbent assay.. RESULTS: All three biomarkers were significantly increased in patients living with HIV-1 from all countries as compared to controls. No differences were found between males and females. Vietnamese and Ethiopian patients had ...

Endotoxin detection in human patients has been a difficult challenge, in part due to the fact that the conserved active portion of the molecule (lipid A) is a relatively small epitope only amenable to binding by a single ligand at any one instance and low levels (pg/ml) are capable of stimulating the immune system. The endotoxin activity assay, a bioassay based on neutrophil activation by complement opsonized immune complexes of lipopolysaccharide (LPS), has allowed the specific detection of the lipid A epitope of LPS in a rapid whole blood assay format. This review summarizes diagnostic studies utilizing the endotoxin activity assay in a variety of hospital patient populations in whom endotoxin is postulated to play a significant role in disease etiology. These include ICU patients at risk of developing sepsis syndrome, abdominal and cardiovascular surgery patients and patients with serious traumatic injury. Significant features of these studies include the high negative predictive value of the assay

TY - JOUR. T1 - Development of a new endotoxin sensor with intermittent injection of limulus reagent for continuous monitoring of dialysate fluid. AU - Aoyagi, S.. AU - Yoshimi, Y.. AU - Sakai, K.. AU - Aketagawa, J.. AU - Tanaka, S.. PY - 1995/1/1. Y1 - 1995/1/1. N2 - This report describes a method of continuously, stably, and inexpensively measuring endotoxin (ET) concentrations in dialysate fluid using an ET sensor with intermittent injection of limulus reagent. An ET solution simulating dialysate fluid was sampled in a single tube at a flow rate of 260 μl/min and mixed with 30 μl of limulus reagent intermittently injected into the tube. The absorbance of the solution was measured after the limulus reaction at 313 or 318° K at 26 min. A good linear relationship (r = 0.98) between peak area of absorbance and ET concentration at ET concentrations ranging from 0 to 0.12 endotoxin unit (EU)/ml was obtained, using a spectrophotometer with a cell volume of 8 μl. The baseline rose after the ...

en] OBJECTIVE: To assess the predictive value of the endotoxin level in the bronchoalveolar lavage (BAL) and to propose to the clinician a guide in the diagnosis of gram-negative bacterial (GNB) pneumonia. DESIGN: Retrospective and prospective studies to investigate the relation between endotoxin level and quantitative bacterial culture of BAL and to test the predictive value of a defined threshold. SETTING: University hospital general intensive care unit. PATIENTS: In the first part of the study, 77 consecutive ventilated patients with clinical suspicion of nosocomial pneumonia between January 1995 and January 1996. In the second part of the study, 93 consecutive ventilated patients studied prospectively between February 1996 and April 1997. MEASUREMENTS AND MAIN RESULTS: Quantitative cultures for aerobic bacteria were performed directly from the fluid. Bacterial species were determined with standard techniques. The detection of endotoxin in BAL was made using a quantitative chromogenic Limulus ...

The objective of this study was to investigate the occurrence of bacteremia in dairy cows with severe mastitis. Milk samples were collected from affected udder quarters, and corresponding blood samples were collected from dairy cows with severe mastitis at the time of diagnosis before any therapeutic measures were undertaken. The cultural detection of pathogens in blood classified a bacteremia. Further diagnostic tests were performed to provide evidence of bacteremia. This was realized by PCR with regard to S. aureus, E. coli and S. uberis and the Limulus test. Detection of culturable pathogens in the blood of cows with severe clinical mastitis was rare and occurred in only one of 70 (1.4%) cases. Overall, bacterial growth was detected in 53 of 70 (75.7%) milk samples. S. uberis (22/70), E. coli (12/70) and S. aureus (4/70) were the most frequently isolated pathogens from milk of cows with severe mastitis. PCR was performed in 38 of 70 (54.3%) blood samples. PCR was positive in eight of 38 ...

The objective of this study was to investigate the occurrence of bacteremia in dairy cows with severe mastitis. Milk samples were collected from affected udder quarters, and corresponding blood samples were collected from dairy cows with severe mastitis at the time of diagnosis before any therapeutic measures were undertaken. The cultural detection of pathogens in blood classified a bacteremia. Further diagnostic tests were performed to provide evidence of bacteremia. This was realized by PCR with regard to S. aureus, E. coli and S. uberis and the Limulus test. Detection of culturable pathogens in the blood of cows with severe clinical mastitis was rare and occurred in only one of 70 (1.4%) cases. Overall, bacterial growth was detected in 53 of 70 (75.7%) milk samples. S. uberis (22/70), E. coli (12/70) and S. aureus (4/70) were the most frequently isolated pathogens from milk of cows with severe mastitis. PCR was performed in 38 of 70 (54.3%) blood samples. PCR was positive in eight of 38 ...

Pyrogenicity is the ability of a chemical agent or other substance to produce a febrile reponse. Pyrogenic responses may be material-mediated, endotoxin-mediated, or mediated by other substances, such as components of gram-positive bacteria and fungi. Method of Pyrogenicity test: Rabbit pyrogen test, TAL test. ...

I am in the beginning stages of an edition of prints Im thinking about calling Limulus. It is going to be similar in lay out and size to a piece I wrote about a few months ago called Archeopteris. Limulus polyphemus is the scientific name for the Horseshoe Crab. I was reading up on it for my design and one scientists argument was that we should not refer to the animal as a Horseshoe Crab because it is not as closely related to the crab family as the name would let on. It is instead more closely related to spiders as it resides in the Arthopod family. My initial intention for creating this print was to honor the longevity of the Limulus. I knew it was an old animal and its physical appearance has remained virtually unchanged through the eons. What I did not know was that the oldest Limulus fossil found to date is roughly 445 million years old. That puts it in the middle of the Ordovician period which is the period directly following the Cambrian period. They have survived 5 major mass ...

This LAL-based assay makes use of the same reaction used for the chromogenic LAL test. Gram-negative organisms, with bacterial endotoxin, initiate the LAL coagulase cascade that results in activation of the proclotting enzyme, a protease. In the LAL test, this enzyme cleaves a peptide from the horseshoe crab coagulen, resulting in a clot. It can also cleave a peptide from a synthetic substrate, yielding a chromophore (p-nitroaniline) which is yellow and can be measured photometrically at 385 nm . Gram-positive organisms, lacking endotoxin, do not trigger the color change in this method, while gram-negative organisms do trigger the colour change and the advantage is that the results are available within 10 minutes. ...

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Get an overview of bacterial endotoxin testing, including the major types of LAL assays used for quality control in pharmaceutical and biomedical manufacturing.

From the hemocyte granules of the horseshoe crabs Limulus and Tachypleus. Factor B is activated by limulus clotting factor C. In peptidase family S1 (trypsin family)

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The horseshoe crab plays an integral role in human health. The LAL compound and the LAL test save humans. Yet, the horseshoe crabs populations are declining.

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In this webinar, Jeffrey Weber from Pfizer discusses options for endotoxin and compendial testing of pharmaceutical products, with a specific emphasis on the Endosafe® endotoxin testing platforms.

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Indicated for replenishing fluid losses, as an energy source and for restoration or maintenance of sodium and chloride ion concentrations. Colourless or white crystal and is freely soluble in water. Nonpyrogenic solutions. Sterile. ...

Page 3 of 4 - Single Cellular --| Multi Cellular? - posted in Best all time threads.: The cells differentiate. There are a few different types of cells found in a sponge, choanocytes and amoebocytes/archaeocytes were just a couple of examples.Genetically? Or do they just take on different tasks but can change to do those?

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Dr Lal PathLabs, one of the largest lab testing companies in India, left a huge cache of patient data on a public server for months, TechCrunch has learned. Australia-based security expert Sami Toivonen found the exposed data and reported it to Dr Lal PathLabs in September. The company quickly shut down access to the bucket but the company did not reply, Toivonen told TechCrunch.

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