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1.
São Paulo; s.n; s.n; 2019. 109 p. graf, tab.
Thesis in Portuguese | LILACS (Americas) | ID: biblio-1022813

ABSTRACT

A fim de garantir a qualidade final de produtos os laboratórios de análise microbiológica fornecem dados sobre a qualidade dos mesmos em todas as suas etapas de produção. A crescente preocupação com a saúde dos pacientes conduz à busca de métodos que forneçam resultados precisos e rápidos, pois possibilitam que ações corretivas sejam tomadas em tempo real. O presente trabalho teve por objetivo avaliar o potencial de tecnologia alternativa no monitoramento de endotoxina bacteriana na água tratada para diálise e dialisato e avaliar o potencial da citometria de fluxo na análise de água. Para isso utilizou-se Portable Test System (PTS®) como método alternativo para detecção de endotoxina bacteriana no monitoramento da água tratada para diálise e dialisato, o qual foi validado frente ao método convencional farmacopeico. Paralelamente realizou-se revisão narrativa da literatura a fim de avaliar a aplicabilidade da citometria de fluxo em análises de água. A análise dos diferentes parâmetros de validação para endotoxina bacteriana no método alternativo mostrou que, exceto para a menor diluição analisada, houve linearidade e precisão nos resultados. Por outro lado a concentração de 0,25 UE/mL foi a menor que apresentou exatidão e especificidade. Observou-se ainda, que o limite de detecção foi de 0,125UE/mL e o de quantificação de 0,25 UE/mL, portanto o intervalo foi de 0,25-1,0 UE/mL. Adicionalmente pela análise de resistência pode-se perceber que ao variar analistas não houve diferença significativa. Em relação ao tempo de análise em uma condição de rotina laboratorial com muitas amostras, o PTS® mostrou-se demorado. Ressalta ainda, que seria importante que a legislação vigente determinasse a análise mensal de endotoxinas no dialisato. A revisão da literatura evidencia o potencial da tecnologia de citometria de fluxo, pois a mesma mostrou-se satisfatória quando comparada a metodologias convencionais para análise de água. O trabalho desenvolvido permitiu concluir que o PTS®) mostrou-se adequado para analisar amostras in loco, permitindo análises em tempo real, que para as quais haja a expectativa de ausência de endotoxinas ou de concentração respeitando o intervalo de 0,25 UE/mL a 1,0 UE/mL. Quanto a citometria de fluxo, esta mostrou-se uma tecnologia promissora em analisar amostras de água, sendo portanto recomendável proceder a estudos de validação e aplicabilidade


In order to guarantee the final quality of products, the microbiological analysis laboratories provide data about their quality at all production stages. The growing concern for patients' health leads to the search for methods that provide accurate and fast results, as they enable corrective actions to be taken in real time. The present work aimed to evaluate the alternative technology potential in the monitoring of bacterial endotoxin in treated water for dialysis and dialysate and to evaluate the potential of flow cytometry in water analysis. The different validation parameters analysis for bacterial endotoxin in alternative method showed that, except for the lowest dilution analyzed, there was linearity and precision in the results. On the other hand, the concentration of 0.25 EU / mL was the lowest that presented accuracy and specificity. It was further observed that the detection limit was 0.125UE / mL and the quantification limit was 0.25 EU / mL, so the range was 0.25-1.0 EU / mL. Additionally by the ruggedness analysis it was possible to perceived that when varying analysts there was no significant difference. Regarding the analysis time in a laboratory routine condition with many samples, the PTS® was was time consuming. It was also observed that it would be important to determine monthly analysis of endotoxins in dialysate. The literature review evidence the flow cytometry technology potential of the because it was satisfactory when compared to conventional methodologies for water analysis. The research showed that the PTS® was suitable for analyzing samples in loco, allowing real-time analyzes, for which there is expectation of endotoxins absence or concentration respecting the range of 0.25 EU / mL to 1.0 EU / mL. For the flow cytometry, it was shown to be a promising technology for analyzing water samples, and it is therefore advisable to carry out validation and applicability studies


Subject(s)
Water Microbiology , Microbiological Techniques/classification , Renal Dialysis , Monitoring , Dialysis , Endotoxins/analysis , Flow Cytometry/instrumentation
2.
Rev. Esc. Enferm. USP ; 50(3): 405-410, June 2016. tab, graf
Article in English | LILACS (Americas) | ID: lil-792770

ABSTRACT

Abstract OBJECTIVE To investigate endotoxins in sterilized surgical instruments used in hip arthroplasties. METHOD A descriptive exploratory study conducted in a public teaching hospital. Six types of surgical instruments were selected, namely: acetabulum rasp, femoral rasp, femoral head remover, chisel box, flexible bone reamer and femoral head test. The selection was based on the analysis of the difficulty in removing bone and blood residues during cleaning. The sample was made up of 60 surgical instruments, which were tested for endotoxins in three different stages. The EndosafeTM Gel-Clot LAL (Limulus Amebocyte Lysate method) was used. RESULT There was consistent gel formation with positive analysis in eight instruments, corresponding to 13.3%, being four femoral rasps and four bone reamers. CONCLUSION Endotoxins in quantity ≥0.125 UE/mL were detected in 13.3% of the instruments tested.


Resumen OBJETIVO Investigar las endotoxinas en instrumentales quirúrgicos esterilizados empleados en artroplastias de la cadera. MÉTODO Estudio exploratorio, descriptivo, desarrollado en un hospital público de enseñanza. Fueron seleccionados seis tipos de instrumentales, a saber: raspa de acetábulo, raspa femoral, sacacorchos (para extraer la cabeza femoral), gubia quirúrgica, fresa femoral y cabeza femoral de prueba. La selección se hizo desde el análisis de la dificultad para la retirada de residuos de sangre y hueso durante la limpieza. La muestra estuvo constituida de 60 instrumentales quirúrgicos, que fueron puestos a prueba para endotoxinas en tres momentos diferentes. Se utilizó el método de gelificación (Gel-clot) Lisado de Amebocitos de Limulus (LAL) Endosafe(tm). RESULTADO Hubo formación de gel consistente con análisis positivo en ocho instrumentos, lo que corresponde a el 13,3%, siendo cuatro raspas femorales y cuatro fresas femorales. CONCLUSIÓN Fueron detectadas endotoxinas en cantidad ≥0,125 UE/mL en el 13,3% de los instrumentos probados.


Resumo OBJETIVO Investigar endotoxinas em instrumentais cirúrgicos esterilizados empregados em artroplastias do quadril. MÉTODO Estudo exploratório, descritivo, desenvolvido em um hospital público de ensino. Foram selecionados seis tipos de instrumentais, a saber: raspa acetabular, raspa femural, saca-cabeça de fêmur, formão box, fresa de fêmur e cabeça de prova de fêmur. A seleção foi feita a partir da análise da dificuldade para a remoção de resíduos de sangue e osso durante a limpeza. A amostra foi constituída por 60 instrumentais cirúrgicos, que foram testados para endotoxinas em três momentos distintos. Foi utilizado o método de gel-clot pelo Limulus Amebócito Lisado (LAL) Endosafe(tm). RESULTADO Houve formação de gel consistente com análise positiva em oito instrumentais, o que corresponde a 13,3%, sendo quatro raspas de fêmur e quatro fresas de fêmur. CONCLUSÃO Foram detectadas endotoxinas em quantidade ≥0,125 UE/mL em 13,3% dos instrumentais testados.


Subject(s)
Arthroplasty, Replacement, Hip/instrumentation , Endotoxins/analysis , Sterilization
3.
J. appl. oral sci ; 23(4): 431-435, July-Aug. 2015. tab, ilus
Article in English | LILACS (Americas) | ID: lil-759357

ABSTRACT

AbstractThe presence of endotoxin inside the root canal has been associated with periapical inflammation, bone resorption and symptomatic conditions.Objectives To determine, in vitro, the effect of QMix® and other three root canal irrigants in reducing the endotoxin content in root canals.Material and Methods Root canals of single-rooted teeth were prepared. Samples were detoxified with Co-60 irradiation and inoculated with E. coli LPS (24 h, at 37°C). After that period, samples were divided into 4 groups, according to the irrigation solution tested: QMix®, 17% EDTA, 2% chlorhexidine solution (CHX), and 3% sodium hypochlorite (NaOCl). LPS quantification was determined by Limulus Amebocyte Lysate (LAL) assay. The initial counting of endotoxins for all samples, and the determination of LPS levels in non-contaminated teeth and in contaminated teeth exposed only to non-pyrogenic water, were used as controls.Results QMix® reduced LPS levels, with a median value of 1.11 endotoxins units (EU)/mL (p<0.001). NaOCl (25.50 EU/mL), chlorhexidine (44.10 EU/mL) and positive control group (26.80 EU/mL) samples had similar results. Higher levels were found with EDTA (176.00 EU/mL) when compared to positive control (p<0.001). There was no significant difference among EDTA, NaOCl and CHX groups. Negative control group (0.005 EU/mL) had statistically significant lower levels of endotoxins when compared to all test groups (p<0.001).Conclusion QMix® decreased LPS levels when compared to the other groups (p<0.001). 3% NaOCl, 2% CHX and 17% EDTA were not able to significantly reduce the root canal endotoxins load.


Subject(s)
Humans , Biguanides/pharmacology , Endotoxins/analysis , Escherichia coli/drug effects , Lipopolysaccharides/analysis , Polymers/pharmacology , Root Canal Irrigants/pharmacology , Analysis of Variance , Chlorhexidine/pharmacology , Dental Pulp Cavity/drug effects , Dental Pulp Cavity/microbiology , Disinfectants/pharmacology , Edetic Acid/pharmacology , Reference Values , Reproducibility of Results , Sodium Hypochlorite/pharmacology , Time Factors
4.
Article in English | WPRIM (Western Pacific) | ID: wprim-86402

ABSTRACT

Hazardous biochemical agents in animal husbandry indoor environments are known to promote the occurrence of various illnesses among workers and animals. The relationship between endotoxin levels in dust collected from chicken farms and various immunological markers was investigated. Peripheral blood was obtained from 20 broiler chickens and 20 laying hens from four different chicken farms in Korea. Concentrations of total or respirable dust in the inside the chicken farm buildings were measured using a polyvinyl chloride membrane filter and mini volume sampler. Endotoxin levels in the dust were determined by the Limulus Amebocyte Lysate Kinetic method. Interferon-gamma production by peripheral blood mononuclear cells stimulated with concanavalin A was significantly lower in broilers or layers from the farms with higher endotoxin concentrations than the chickens from the farms with lower endotoxin levels. An opposite pattern was observed for plasma cortisol concentrations with higher cortisol levels found in chickens from the farms with higher endotoxin levels. When peripheral lymphocytes were examined, the percentage of CD3-Ia+ B cells was lower in layers from farms with higher endotoxin levels than those from locations with lower endotoxin levels. Overall, these results suggest a probable negative association between dust endotoxin levels and cell-mediated immunity in chickens.


Subject(s)
Animal Husbandry , Animals , Biomarkers/blood , Chickens/immunology , Dust/analysis , Endotoxins/analysis , Housing, Animal , Immunity, Cellular
5.
Acta cir. bras ; 29(6): 359-364, 06/2014. graf
Article in English | LILACS (Americas) | ID: lil-711592

ABSTRACT

PURPOSE: To determine the role of mesenteric lymph reperfusion (MLR) on endotoxin translocation in brain to discuss the mechanism of brain injury subjected to superior mesenteric artery occlusion (SMAO) shock. METHODS: Twenty-four rats were randomly assigned to MLR, SMAO, MLR+SMAO and sham groups. MLR was performed by clamping the mesenteric lymph duct (MLD) for 1 h and then allowing reperfusion for 2 h in the MLR group; SMAO involved clamping the superior mesenteric artery (SMA) for 1 h, followed by reperfusion for 2 h in the SMAO group; occlusion of both the SMA and MLD for 1 h was followed by reperfusion for 2 h in the MLR+SMAO group rats. RESULTS: SMAO shock induced severe increased levels of the endotoxin, lipopolysaccharide receptor, lipopolysaccharide-binding protein, intercellular adhesion molecule-1 and tumor necrosis factor-α. Concurrently, MLR after SMAO shock further aggravates these deleterious effects. CONCLUSION: Mesenteric lymph reperfusion exacerbated the endotoxin translocation in brain; thereby increased inflammatory response occurred, suggesting that the intestinal lymph pathway plays an important role in the brain injury after superior mesenteric artery occlusion shock. .


Subject(s)
Animals , Male , Bacterial Translocation/physiology , Brain Injuries/etiology , Endotoxins/physiology , Lymphatic Vessels/physiology , Mesentery , Mesenteric Vascular Occlusion/physiopathology , Reperfusion Injury/physiopathology , Acute-Phase Proteins/analysis , /analysis , Brain Injuries/metabolism , Carrier Proteins/analysis , Disease Models, Animal , Enzyme-Linked Immunosorbent Assay , Endotoxins/analysis , Intercellular Adhesion Molecule-1/analysis , Ligation , Lymphatic Vessels/surgery , Mesenteric Artery, Superior , Membrane Glycoproteins/analysis , Mesenteric Vascular Occlusion/complications , Random Allocation , Rats, Wistar , Reperfusion Injury/complications , Time Factors , Tumor Necrosis Factor-alpha/analysis
6.
Braz. j. med. biol. res ; 47(5): 376-383, 02/05/2014. graf
Article in English | LILACS (Americas) | ID: lil-709439

ABSTRACT

The intestinal lymph pathway plays an important role in the pathogenesis of organ injury following superior mesenteric artery occlusion (SMAO) shock. We hypothesized that mesenteric lymph reperfusion (MLR) is a major cause of spleen injury after SMAO shock. To test this hypothesis, SMAO shock was induced in Wistar rats by clamping the superior mesenteric artery (SMA) for 1 h, followed by reperfusion for 2 h. Similarly, MLR was performed by clamping the mesenteric lymph duct (MLD) for 1 h, followed by reperfusion for 2 h. In the MLR+SMAO group rats, both the SMA and MLD were clamped and then released for reperfusion for 2 h. SMAO shock alone elicited: 1) splenic structure injury, 2) increased levels of malondialdehyde, nitric oxide (NO), intercellular adhesion molecule-1, endotoxin, lipopolysaccharide receptor (CD14), lipopolysaccharide-binding protein, and tumor necrosis factor-α, 3) enhanced activities of NO synthase and myeloperoxidase, and 4) decreased activities of superoxide dismutase and ATPase. MLR following SMAO shock further aggravated these deleterious effects. We conclude that MLR exacerbates spleen injury caused by SMAO shock, which itself is associated with oxidative stress, excessive release of NO, recruitment of polymorphonuclear neutrophils, endotoxin translocation, and enhanced inflammatory responses.


Subject(s)
Animals , Male , Lymph/metabolism , Mesenteric Vascular Occlusion/complications , Reperfusion Injury/etiology , Reperfusion/adverse effects , Spleen/injuries , Acute-Phase Proteins/analysis , Adenosine Triphosphatases/analysis , /analysis , Carrier Proteins/analysis , Endotoxins/analysis , Intercellular Adhesion Molecule-1/analysis , Intestines/blood supply , Mesenteric Artery, Superior , Malondialdehyde/analysis , Membrane Glycoproteins/analysis , Nitric Oxide Synthase/analysis , Nitric Oxide/analysis , Peroxidase/analysis , Rats, Wistar , Spleen/pathology , Superoxide Dismutase/analysis , Tumor Necrosis Factor-alpha/analysis
7.
J. appl. oral sci ; 21(1): 25-31, 2013. tab
Article in English | LILACS (Americas), BBO | ID: lil-684991

ABSTRACT

The purpose of this study was to evaluate the effectiveness of glycolic propolis (PRO) and ginger (GIN) extracts, calcium hydroxide (CH), chlorhexidine (CLX) gel and their combinations as ICMs (ICMs) against Candida albicans,Enterococcus faecalis, Escherichia coli and endotoxins in root canals. Material and Methods: After 28 days of contamination with microorganisms, the canals were instrumented and then divided according to the ICM: CH+saline; CLX, CH+CLX, PRO, PRO+CH; GIN; GIN+CH; saline. The antimicrobial activity and quantification of endotoxins by the chromogenic test of Limulus amebocyte lysate were evaluated after contamination and instrumentation at 14 days of ICM application and 7 days after ICM removal. Results and Conclusion: After analysis of results and application of the Kruskal-Wallis and Dunn statistical tests at 5% significance level, it was concluded that all ICMs were able to eliminate the microorganisms in the root canals and reduce their amount of endotoxins; however, CH was more effective in neutralizing endotoxins and less effective against C. albicans and E. faecalis, requiring the use of medication combinations to obtain higher success.


Subject(s)
Humans , Candida albicans/drug effects , Dental Pulp Cavity/microbiology , Endotoxins/analysis , Enterococcus faecalis/drug effects , Escherichia coli/drug effects , Ginger/chemistry , Propolis/pharmacology , Calcium Hydroxide/pharmacology , Chlorhexidine/pharmacology , Disinfectants/pharmacology , Endotoxins/chemistry , Root Canal Preparation , Root Canal Irrigants/pharmacology , Statistics, Nonparametric , Time Factors
8.
J. appl. oral sci ; 20(6): 661-666, Nov.-Dec. 2012. tab
Article in English | LILACS (Americas) | ID: lil-660638

ABSTRACT

Gram-negative bacteria play an essential role in endodontic infections because they have virulence factors such as endotoxin. Due to its potential cytotoxic activity, special attention has been given to the removal/neutralization of this endotoxin in the root canal system. OBJECTIVE: The aim of this study was to evaluate the influence of the apical enlargement size (AES) by using rotary instruments on the endotoxin level reduction of dental root canals. MATERIAL AND METHODS: Forty root canals of the mandibular premolar teeth were used. Escherichia coli endotoxin (055: B55) was inoculated into thirty root canals. Ten teeth served as the negative control group. After the incubation period, the first endotoxin samples were collected from the root canals with a sterile/apyrogenic paper point for the analysis of the endotoxin units (EU/mL) present before instrumentation (S1). Specimen instrumentation was performed with the Mtwo® rotary system in the sequence 10/.04, 15/.05, 20/.06, 25/.06, 30/.05, 35/.04 and 40/.04. To monitor the effectiveness of increasing apical enlargement on endotoxin removal, the second endotoxin samples were collected from all the root canals after instrumentation with the following instruments: #25/.06- (S2); #30/.05- (S3); # 35/.04- (S4); and #40/.04- (S5). Limulus amebocyte lysate (LAL) was used to quantify the levels of endotoxin. The results were statistically compared by using repeated measures of ANOVA with post hoc Tukey testing. RESULTS: Increasing levels of endotoxin removal was achieved by large sized apical enlargement: S2 (AES #25/.06)- 89.2%, S3 (AES #30/.05)- 95.9%, S4 (AES #35/.04)- 97.8% and S5 (AES #40/.04)- 98.2%. Substantial reduction of endotoxin content was obtained in S4 and S5 compared to S2 (p<0.05), however, the root canal preparation was not able to eliminate the endotoxin. CONCLUSIONS: Under the conditions of this study, it was concluded that the reduction of endotoxin levels of the dental root canals could be predicted by increasing the apical enlargement size.


Subject(s)
Humans , Dental Instruments , Endotoxins/analysis , Root Canal Preparation/instrumentation , Tooth Apex , Analysis of Variance , Nickel , Odontometry/instrumentation , Random Allocation , Statistics, Nonparametric , Time Factors , Titanium
9.
J. appl. oral sci ; 19(2): 106-112, May-Apr. 2011. tab
Article in English | LILACS (Americas) | ID: lil-586030

ABSTRACT

OBJECTIVE: The purpose of this study was to evaluate the effcacy of auxiliary chemical substances and intracanal medications on Escherichia coli and its endotoxin in root canals. MATERIAL AND METHODS: Teeth were contaminated with a suspension of E. coli for 14 days and divided into 3 groups according to the auxiliary chemical substance used: G1) 2.5 percent sodium hypochlorite (NaOCl); G2) 2 percent chlorhexidine gel (CLX); G3) pyrogenfree solution. After, these groups were subdivided according to the intracanal medication (ICM): A) Calcium hydroxide paste (Calen®), B) polymyxin B, and C) Calcium hydroxide paste+2 percent CLX gel. For the control group (G4), pyrogen-free saline solution was used without application of intracanal medication. Samples of the root canal content were collected immediately after biomechanical preparation (BMP), at 7 days after BMP, after 14 days of intracanal medication activity, and 7 days after removal of intracanal medication. The following aspects were evaluated for all collections: a) antimicrobial activity; b) quantifcation of endotoxin by the Limulus Amebocyte Lysate test (LAL). Results were analyzed by the Kruskal-Wallis and Dunn's tests at 5 percent signifcance level. RESULTS: The 2.5 percent NaOCl and CLX were able to eliminate E. coli from root canal lumen and reduced the amount of endotoxin compared to saline. CONCLUSIONS: It was concluded that 2.5 percent NaOCl and CLX were effective in eliminating E. coli. Only the studied intracanal medications were to reduce the amount of endotoxin present in the root canals, regardless of the irrigant used.


Subject(s)
Humans , Dental Pulp Cavity/microbiology , Endotoxins/analysis , Escherichia coli/drug effects , In Vitro Techniques , Root Canal Irrigants/pharmacology , Anti-Bacterial Agents/pharmacology , Calcium Hydroxide/pharmacology , Chlorhexidine/pharmacology , Dental Pulp Cavity/chemistry , Polymyxin B/pharmacology , Root Canal Preparation , Statistics, Nonparametric , Sodium Hypochlorite/pharmacology
10.
Braz. j. pharm. sci ; 46(3): 551-554, July-Sept. 2010. tab
Article in English | LILACS (Americas) | ID: lil-564922

ABSTRACT

F-18 FDG (2-[18-F] fluoro-2-deoxy-D-glucose) is the most frequently used radiopharmaceutical for PET and PET CT imaging exams. The FDA recently approved the use of the PTS TM (Portable Test System) as an alternative to the standard test proposed by the United States Pharmacopeia using the LAL (Limulus Amebocyte Lysates), that takes longer to perform (about 1h) than the PTS TM (15 min). Recent studies have demonstrated that radiation could interfere with the PTS TM test. In order to study the effects of radiation on the PTS TM test and/or equipment, 27 batches of F-18 FDG produced in the Nuclear Engineering Institute were analyzed. The results showed that no direct correlation with radiation was found in any of the cases.


O FDG-18 é o radiofármaco mais utilizado nos exames de PET e PET CT. O FDA recentemente aprovou o uso do PTS TM (Portable Test System) como método alternativo ao teste padrão de endotoxina, proposto pela Farmacopéia Americana, considerando que no primeiro há um tempo de espera de 1 hora frente a somente 15 minutos do segundo. Estudo recentes demonstram que a radiação poderia interferir no teste do PTS TM. De modo a avaliar os efeitos da radiação no teste PTS TM foram analisados 27 lotes de F-18 FDG produzidos no Instituto de Engenharia Nuclear. Os resultados demonstraram que em todos os casos nenhuma correlação direta com a radiação foi observada.


Subject(s)
Endotoxins/chemistry , Research/methods , Radiation , Endotoxins/analysis , Nuclear Medicine , Radiopharmaceuticals
11.
Rev. bras. cardiol. invasiva ; 17(2): 234-238, abr.-jun. 2009. tab, graf
Article in Portuguese | LILACS (Americas) | ID: lil-527898

ABSTRACT

A reação pirogênica é desencadeada pela presença, na corrente sanguínea, de soluções contaminadas, infundidas no paciente, contendo endotoxinas ou produtos de degradação proteica. Apesar da baixa incidência, seu foco deve ser investigado e erradicado, pois essas ocorrências podem ser deletérias e refletem a qualidade do serviço prestado, já que a causa pode ser proveniente de fatores relacionados ao exame, às soluções utilizadas ou ao processo de esterilização...


Subject(s)
Humans , Male , Female , Catheterization/adverse effects , Catheterization , Endotoxins/analysis , Water Microbiology/standards , Pyrogens/analysis , Pyrogens/adverse effects , Equipment Contamination , Water Purification/methods , Equipment Reuse/standards
12.
P. R. health sci. j ; 27(1): 21-26, Mar. 2008.
Article in English | LILACS (Americas) | ID: lil-491620

ABSTRACT

Consumers increasingly use bottled water and home water treatment systems to avoid direct tap water. According to the International Bottled Water Association (IBWA), an industry trade group, 5 billion gallons of bottled water were consumed by North Americans in 2001. The principal aim of this study was to assess the microbial quality of in-house and imported bottled water for human consumption, by measurement and comparison of the concentration of bacterial endotoxin and standard cultivable methods of indicator microorganisms, specifically, heterotrophic and fecal coliform plate counts. A total of 21 brands of commercial bottled water, consisting of 10 imported and 11 in-house brands, selected at random from 96 brands that are consumed in Puerto Rico, were tested at three different time intervals. The Standard Limulus Amebocyte Lysate test, gel clot method, was used to measure the endotoxin concentrations. The minimum endotoxin concentration in 63 water samples was less than 0.0625 EU/mL, while the maximum was 32 EU/mL. The minimum bacterial count showed no growth, while the maximum was 7,500 CFU/mL. Bacterial isolates like P. fluorescens, Corynebacterium sp. J-K, S. paucimobilis, P. versicularis, A. baumannii, P. chlororaphis, F. indologenes, A. faecalis and P. cepacia were identified. Repeated measures analysis of variance demonstrated that endotoxin concentration did not change over time, while there was a statistically significant (p < 0.05) decrease in bacterial count over time. In addition, multiple linear regression analysis demonstrated that a unit change in the concentration of endotoxin across time was associated with a significant (p < 0.05) reduction in the bacteriological cell count. This analysis evidenced a significant time effect in the average log bacteriological cell count. Although bacterial growth was not detected in some water samples, endotoxin was present. Measurement of Gram-negative bacterial endotoxins is one of the methods that have...


Subject(s)
Water Supply/analysis , Endotoxins/analysis , Enterobacteriaceae/isolation & purification , Water Microbiology , Colony Count, Microbial , Feces/microbiology
13.
Article in English | WPRIM (Western Pacific) | ID: wprim-96035

ABSTRACT

Protozoan parasites of the genus Leishmania cause a number of important human diseases. One of the key determinants of parasite infectivity and survival is the surface glycoconjugate lipophosphoglycan (LPG). In addition, LPG is shown to be useful as a transmission blocking vaccine. Since culture supernatant of parasite promastigotes is a good source of LPG, we made attempts to characterize functions of the culture supernatant, and membrane LPG isolated from metacyclic promastigotes of Leishmania major. The purification scheme included anion-exchange chromatography, hydrophobic interaction chromatography and cold methanol precipitation. The purity of supernatant LPG (sLPG) and membrane LPG (mLPG) was determined by SDS-PAGE and thin layer chromatography. The effect of mLPG and sLPG on nitric oxide (NO) production by murine macrophages cell line (J774.1A) was studied. Both sLPG and mLPG induced NO production in a dose dependent manner but sLPG induced significantly higher amount of NO than mLPG. Our results show that sLPG is able to promote NO production by murine macrophages.


Subject(s)
Nitric Oxide/analysis , Mice, Inbred BALB C , Mice , Macrophages/drug effects , Leishmania major/chemistry , Glycosphingolipids/isolation & purification , Endotoxins/analysis , Electrophoresis, Polyacrylamide Gel , Culture Media , Chromatography, Thin Layer/methods , Cell Membrane/chemistry , Cell Line , Animals
14.
Acta cient. venez ; 56(4): 139-148, 2005. graf
Article in Spanish | LILACS (Americas) | ID: lil-537158

ABSTRACT

Bajo la definición de malestar general se agrupan una constelación de signos y síntomas tales como fiebre, disminución de la actividad, pérdida del apetito, disminución de la libido, trastornos del sueño y modificaciones en el umbral del dolor, entre otros. Estas manifestaciones son necesarias para hacer frente a procesos infecciosos y son iniciadas por el sistema inmunitario y controladas por el sistema nervioso, por lo que representan un ejemplo ideal de interacción entre ambos componentes. En el presente estudio se analizaron los cambios conductuales inducidos en el umbral del dolor luego de la administración i.p. de lipopolisacárido (LPS), una endotoxina que forma parte de la pared de bacterias gram negativas. Adicionalmente, se determinaron los cambios que ocurren en los patrones de descarga de dos poblaciones de neuronas de la formación reticular bulbar, las células “off” y “on”, luego de la administración de la endotoxina. Estas células aparentemente constituyen los brazos ejecutores del sistema de modulación endógena del dolor. Los resultados obtenidos indican que dependiendo de la dosis de LPS empleada (100 o 200 μg/kg) se pueden activar diferentes mecanismos de modulación, que a su vez parecen depender de las dos poblaciones de neuronas bulbares. La dosis de 100 μg/kg de LPS produjo antinocicepción, la cual estuvo asociada a incrementos en la actividad de las células “off” y a la disminución de la actividad de las “on”. La dosis de 200 μg/kg de LPS produjo hiperalgesia, la cual se acompañó de una disminución de la actividad de las celulas off”y un aumento en la actividad de las células on.


The definition of sickness/illness behavior involves a constellation of signs and symptoms such as fever, decrease of physical activity, anorexia, decrease of libido, insomnia and changes of the pain threshold, among others. These manifestations are necessary to face infectious processes and are apparently initiated and controlled by the immune and the nervous system, so that they represent an ideal example of interaction between both components. In the present study the behavioral changes in the pain threshold induced by the i.p. administration of lipopolysaccharide, an endotoxin derived from gram-negative bacterial cell walls, were analyzed. Additionally, changes in the discharge pattern of two neuronal populations of the medullary reticular formation, the off- and on-cells, which apparently act as the executory arm of the endogenous pain modulatory system were determined after endotoxin administration. The results indicate that depending on the LPS dose (100 or 200 μg/kg) different modulatory mechanisms might be activated, which in turn seem to be the consequence of the activation of the two different classes of medullary neurons. Antinociception was produced after administration of 100 μg/kg of LPS. That was associated to an increase of the off-cell firing and a reduction of the on-cell activity. Hyperalgesia was produced after administration of 200 μg/kg of LPS. That effect was accompanied by an inhibition of the off-cell activity and an increase of the on-cell firing. These findings suggest that changes of the pain threshold that occur during sickness/illness involve the differential participation of the two control options of the endogenous pain modulatory system and of specialized mechanisms of communication between the immune and the nervous system.


Subject(s)
Analgesics , Pain/classification , Pain/therapy , Endotoxins/analysis , Hyperalgesia/classification , Lipopolysaccharides/analysis , Behavioral Symptoms/therapy , Nervous System Physiological Phenomena , Neurophysiology
15.
Yonsei Medical Journal ; : 268-274, 2005.
Article in English | WPRIM (Western Pacific) | ID: wprim-99091

ABSTRACT

Based on the biological activity of endotoxin, we propose a possible new method for detecting endotoxin using a pH- indication system of macrophage culture media. After RAW 264.7 macrophage cells were treated with lipopolysaccharide (LPS), the addition of fluorescein to the LPS-treated media reproductively reduced its absorption and emission spectra (it was a dose-dependent reduction). The advantages of this LPS- detection method were compared with the Limulus Amebocyte Lysate (LAL) test by using purified bacterial LPS (Salmonella minnessota, Escherichia coli, and Pseudomonas aeruginosa). Additionally, the absorption and fluorescence intensity of fluorescein, following treatment of RAW 264.7 cells with a high concentration of Staphylococcus aureus (Gram-positive, lysed bacteria), could not generally be detected by the LAL test, but they were found to be reduced, in a dose-response relationship, with this new system. The macrophage culture system-method might be a good supplement to the LAL assay for detection of LPS, Gram-negative and Gram-positive bacteria.


Subject(s)
Animals , Biological Assay/methods , Cells, Cultured , Comparative Study , Contrast Media , Culture Media/chemistry , Endotoxins/analysis , Fluorescein , Hydrogen-Ion Concentration , Limulus Test , Lipopolysaccharides/analysis , Macrophages/chemistry , Mice
16.
Article in Ml | WPRIM (Western Pacific) | ID: wprim-629947

ABSTRACT

The aim of this study was to evaluate the morphological and biological properties of a locally produced "Bovine Bone Sponge" for use in dentistry. Bovine bone sponge was prepared from local calf bone. Endotoxin level and surface properties were investigated. The pore size and water uptake ability were measured and results were compared with the commercial haemostatic agent. The material was tested for its haemostatic property and its inhibition of alveolar bone resorption in a sheep model following dental extraction. Results revealed a significant difference in haemostatic effect, and a shorter bleeding time and a lower rate of alveolar bone resorption in bovine bone sponge compare to a commercial haemostatic agent.


Subject(s)
Alveolar Bone Loss/prevention & control , Biocompatible Materials , Bone and Bones , Endotoxins/analysis , Hemorrhage/therapy , Hemostatics , Limulus Test , Microscopy, Electron, Scanning , Sheep , Surgical Sponges , Tooth Extraction
17.
Pakistan Journal of Pharmaceutical Sciences. 2004; 17 (1): 89-4
in English | IMEMR (Eastern Mediterranean) | ID: emr-68041

ABSTRACT

Screening of twenty five large volume parenterals including dextrose, electrolytes, mannitol, metronidazole infusions, haemodialysis solution, water for injections and distilled water for the determination of pyrogen using Limulus Amebocyte Lysate [LAL] test has been carried out. Out of different preparations only one metronidazole injection exhibited positive LAL test, which was found pyrogen free with USP rabbit pyrogen test


Subject(s)
Endotoxins/analysis , Limulus Test/methods , Infusions, Parenteral , Rabbits
18.
Scientific Journal of Al-Azhar Medical Faculty [Girls] [The]. 2000; 21 (3): 465-473
in English | IMEMR (Eastern Mediterranean) | ID: emr-55600
19.
Bogotá; s.n; jul. 1999. 180 p. ilus, tab, graf.
Thesis in Spanish | LILACS (Americas) | ID: lil-278170

ABSTRACT

Se valoró la presencia de pirógenos en el antisuero contra venenos de serpientes de los genéros Bothrops, Crotalus y Lachesis elaborado por la planta de producción de sueros antiofídico del Instituto Nacional de Salud en Bogotá Colombia. Se evaluó materia prima, proceso y producto terminado utilizando las técnicas de pirógenos en conejos USP y limulus amebocito lisado (LAL). Los resultados por los dos métodos de análisis para los cinco lotes de materia prima fueron negativos a pirógenos y positivos para los ocho lotes analizados de producto terminado en los que se determinó por LAL niveles de endotoxina entre >13.87 UE/ml y > 24.7 UE/ml; superiores al permitido por la Farmacopea Americana USP XXIII, que fija un valor máximo de 1.67 UE/ml para este tipo de producto. Se estandarizó el uso de la técnica del LAL en el suero de origen equino utilizado como materia prima, el cual presenta inhibidores naturales para la técnica; mediante pretratamientos al plasma para la extracción de inhibidores aplicando diferente técnicas, con los siguientes resultados de mayor a menor capacidad de detección de endotoxina: técnica de sulfato de amonio hasta 0.125 UE/ml; técnica de dilución calor 0.21 UE/ml; técnica de cloroformo dilución 0.42 UE/ml y la técnica con ajuste de pH no detécto endotoxina en la muestra. Se valoró la potencialización de las muestras sometidas a extracción, con la adición de iones mono y divalentes. Se ensayaron los iones NaCl, MnCl, CaCl subíndice 2 y MgCl subíndice 2, en concentraciones de 0.02, 0.05, 0.08M. El mejor resultado se obtuvo con los iones MgCl subíndice 2 y CaCl subíndice 2 a 0.05M, asociado a la técnica de extracción de cloroformo y dilución, en la que aumentó la detección de endotoxina de 0.42 a 0.125 UE/ml con MgCl subíndice 2 y de 0.42 a 0.35 UE/ml con CaCl subíndice 2. Se concluyó que la técnica del LAL ajustada con la técnica sulfato de amonio y calor y la adición MgCL subíndice 2 a 0.05M, sirve como prueba de control para ser aplicada a la materia prima. En el estudio se demostró que durante el proceso de purificación de las inmunoglobulinas, el producto se contamina con endotoxinas bacterianas; fueron responsables de esta contaminación 11.44 por ciento de los materiales, 51.17 por ciento de los equipos y 2.35 por ciento de los reactivos utilizados. Se detecto endotoxinas en 64.96 por ciento de las mustras analizadas. La aplicación del método del LAL en la línea de proceso permitió...


Subject(s)
Antivenins/analysis , Academic Dissertations as Topic , Endotoxins/analysis , Quality Control , Limulus Test , Pyrogens/isolation & purification
20.
Indian J Exp Biol ; 1998 Sep; 36(9): 867-74
Article in English | IMSEAR (South-East Asia), GHL | ID: sea-56154

ABSTRACT

During mammalian preimplantation development, a substantial numbers of embryos are believed to be lost for reasons that are unclear. Using female rats, we investigated whether the developmental status of embryos is influenced by bacterial infection and endotoxin in the reproductive tract. From the vagina of cycling rats (n = 11), 21 bacterial isolates were identified; they were Streptococcus faecalis (S. faecalis; 38%), Escherichia coli (E. coli; 19%), Acinetobactor calcoaceticus (A. calcoaceticus; 14%), and coagulase negative staphylococcus (14%), Micrococcus sp. (5%), Bacillus subtilis (B. subtilis; 5%) and Proteus vulgaris (P. vulgaris; 5%). From the vagina of day 4 pregnant rats (n = 12), 26 isolates were identified; they were S. faecalis (23%), A. calcoaceticus (23%), E. coli (15%), Micrococcus sp. (15%), B. subtilis (8%), P. vulgaris (4%), Staphylococcus aureus (4%), beta-hemolytic streptococcus (4%) and Pseudomonas aeruginosa (4%). Gram negative bacteria found in the vagina of cycling and day 4 pregnant rats were 38% and 46%, respectively. In both, bacterial load was 10(3)-10(5) colony forming units and there was no association with the abnormality of the recovered embryos. However, in two day 4 pregnant animals, pathogenic bacteria (Staphylococcus aureus and beta-hemolytic streptococcus) were isolated and embryos recovered from them were degenerated and deformed. The vagina of day 9 pregnant animals (n = 7) were, however, sterile. Consistently, in all animals, the upper reproductive tract (uterus and oviduct) was devoid of any bacteria and no anaerobic bacteria were isolated from any part of the tract. The levels of endotoxin in the vagina of cycling and day 4 pregnant rats were 1.35 +/- 0.1 and 1.17 +/- 0.1 endotoxin units (EU), respectively. It was undetectable in the oviduct and uterus of all animals (n = 5) except one which showed high levels of endotoxin in uterus (4.5 EU) and oviduct (2.2 EU) and the animal also produced degenerated and deformed embryos. These results indicate that common bacterial flora of vagina may not affect embryo development and the presence of pathogenic bacteria in the vagina and/or endotoxin in reproductive tract could be detrimental to viability of gametes and preimplantation embryos in rats.


Subject(s)
Animals , Bacterial Infections/complications , Embryonic Development , Embryonic and Fetal Development , Endotoxins/analysis , Female , Genitalia, Female/microbiology , Pregnancy , Pregnancy Complications, Infectious/microbiology , Rats , Rats, Wistar
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