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1.
Animals (Basel) ; 14(13)2024 Jul 02.
Artículo en Inglés | MEDLINE | ID: mdl-38998066

RESUMEN

Considering that certain catabolic products of anaerobic chlorophyll degradation inhibit efflux pump activity, this study was conducted to test if feeding pigs a water-soluble chlorophyllin product could affect the antibiotic resistance profiles of select wild-type populations of fecal bacteria. Trial 1 evaluated the effects of chlorophyllin supplementation (300 mg/meal) on fecal E. coli and enterococcal populations in pigs fed twice daily diets supplemented without or with ASP 250 (containing chlortetracycline, sulfamethazine and penicillin at 100, 100 and 50 g/ton, respectively). Trial 2, conducted similarly, evaluated chlorophyllin supplementation in pigs fed diets supplemented with or without 100 g tylosin/ton. Each trial lasted 12 days, and fecal samples were collected and selectively cultured at 4-day intervals to enumerate the total numbers of E. coli and enterococci as well as populations of these bacteria phenotypically capable of growing in the presence of the fed antibiotics. Performance results from both studies revealed no adverse effect (p > 0.05) of chlorophyllin, antibiotic or their combined supplementation on average daily feed intake or average daily gain, although the daily fed intake tended to be lower (p = 0.053) for pigs fed diets supplemented with tylosin than those fed diets without tylosin. The results from trial 1 showed that the ASP 250-medicated diets, whether without or with chlorophyllin supplementation, supported higher (p < 0.05) fecal E. coli populations than the non-medicated diets. Enterococcal populations, however, were lower, albeit marginally and not necessarily significantly, in feces from pigs fed the ASP 250-medicated diet than those fed the non-medicated diet. Results from trial 2 likewise revealed an increase (p < 0.05) in E. coli and, to a lesser extent, enterococcal populations in feces collected from pigs fed the tylosin-medicated diet compared with those fed the non-medicated diet. Evidence indicated that the E. coli and enterococcal populations in trial 1 were generally insensitive to penicillin or chlortetracycline, as there were no differences between populations recovered without or with antibiotic selection. Conversely, a treatment x day of treatment interaction observed in trial 2 (p < 0.05) provided evidence, albeit slight, of an enrichment of tylosin-insensitive enterococci in feces from the pigs fed the tylosin-medicated but not the non-medicated diet. Under the conditions of the present study, it is unlikely that chlorophyllin-derived efflux pump inhibitors potentially present in the chlorophyllin-fed pigs were able to enhance the efficacy of the available antibiotics. However, further research specifically designed to optimize chlorophyll administration could potentially lead to practical applications for the swine industry.

2.
J Thromb Haemost ; 21(3): 639-651, 2023 03.
Artículo en Inglés | MEDLINE | ID: mdl-36696221

RESUMEN

BACKGROUND: Tissue factor pathway inhibitor (TFPI) is the primary inhibitor of events initiating the blood coagulation pathway. Tfpi-/- mice die during embryonic development. The absence of protease-activated receptor (PAR) 4, the major thrombin receptor on mouse platelets, rescues Tfpi-/-mice to adulthood. Among the 3 TFPI isoforms in mice, TFPIα is the only isoform within platelets (pltTFPIα) and the only isoform that inhibits prothrombinase, the enzymatic complex that converts prothrombin to thrombin. OBJECTIVES: To determine biological functions of pltTFPIα. METHODS: Tfpi-/-/Par4-/- mice were irradiated and transplanted with bone marrow from mice lacking or containing pltTFPIα. Thus, PAR4 expression was restored in the recipient mice, which differed selectively by the presence or absence of pltTFPIα and lacked other forms of TFPI. RESULTS: Recipient mice lacking pltTFPIα had reduced survival over the 200-day posttransplant period. Necropsy revealed radiation injury associated with large intraventricular platelet-rich thrombi, whereas other organs were not affected. Thrombi were associated with fibrotic presentations, including increased collagen deposition, periostin-positive activated fibroblasts, myofibroblasts, and macrophage infiltrates. Recipient mice containing pltTFPIα showed evidence of radiation injury but lacked heart pathology. CONCLUSIONS: Tfpi-/-/Par4-/- mice develop severe cardiac fibrosis following irradiation and transplantation with bone marrow lacking pltTFPIα. This pathology is markedly reduced when the mice are transplanted with bone marrow containing pltTFPIα. Thus, in this model system pltTFPIα has an important physiological role in dampening pathological responses mediated by activated platelets within the heart tissue.


Asunto(s)
Plaquetas , Trombosis , Ratones , Animales , Plaquetas/metabolismo , Trombosis/metabolismo , Trombina/metabolismo , Isoformas de Proteínas , Fibrosis
3.
Sci Data ; 8(1): 76, 2021 03 05.
Artículo en Inglés | MEDLINE | ID: mdl-33674581

RESUMEN

The Buzzards Bay Coalition's Baywatchers Monitoring Program (Baywatchers) collected summertime water quality information at more than 150 stations around Buzzards Bay, Massachusetts from 1992 to 2018. Baywatchers documents nutrient-related water quality and the effects of nitrogen pollution. The large majority of stations are located in sub-estuaries of the main Bay, although stations in central Buzzards Bay and Vineyard Sound were added beginning in 2007. Measurements include temperature, salinity, Secchi depth and concentrations of dissolved oxygen, ammonium, nitrate + nitrite, total dissolved nitrogen, particulate organic nitrogen, particulate organic carbon, ortho-phosphate, chlorophyll a, pheophytin a, and in lower salinity waters, total phosphorus and dissolved organic carbon. The Baywatchers dataset provides a long-term record of the water quality of Buzzards Bay and its sub-estuaries. The data have been used to identify impaired waters, evaluate discharge permits, support the development of nitrogen total maximum daily loads, develop strategies for reducing nitrogen inputs, and increase public awareness and generate support for management actions to control nutrient pollution and improve water quality.

4.
Transfusion ; 61(5): 1600-1608, 2021 05.
Artículo en Inglés | MEDLINE | ID: mdl-33580978

RESUMEN

BACKGROUND: Beta-lactam antibiotics are a relatively common cause of immune thrombocytopenia. Because the many beta-lactam drugs now in clinical use have structural similarities, when a patient experiences this complication the question of whether an alternative member of this drug family can safely be used often arises but there are little data available to guide this decision. STUDY DESIGN AND METHODS: Drug-dependent, platelet-reactive antibodies from 32 patients who experienced thrombocytopenia while being treated with a beta-lactam drug of the penam (piperacillin, etc.) or cephem (ceftriaxone etc.) groups were studied for serologic cross-reactivity with other drugs from these families using flow cytometry. Cross-reactions observed were analyzed for correlations with structural features of the drugs tested. RESULTS: Among 14 antibodies specific for penam drugs, five "strong" cross-reactions with other penam drugs were found. Among 18 antibodies specific for cephem drugs, 8 "strong cross-reactions were identified. Antibodies induced by penam drugs did not cross-react strongly with cephem drugs and vice versa. A strong correlation between cross-reactions and similar or identical R1 side groups of the beta-lactams studied was observed. DISCUSSION: The findings suggest that patients who experience immune thrombocytopenia while being treated with a beta-lactam of the penam group can safely be treated with a cephem drug and vice versa. If a patient is to be switched to another beta lactam within the same group, the likelihood of serologic cross-reactivity can be minimized by choosing an agent with a distinctly different R1 side group.


Asunto(s)
Antibacterianos/efectos adversos , Púrpura Trombocitopénica Idiopática/inducido químicamente , beta-Lactamas/efectos adversos , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Antibacterianos/inmunología , Anticuerpos/sangre , Anticuerpos/inmunología , Reacciones Cruzadas , Femenino , Humanos , Masculino , Persona de Mediana Edad , Púrpura Trombocitopénica Idiopática/sangre , Púrpura Trombocitopénica Idiopática/inmunología , Adulto Joven , beta-Lactamas/inmunología
5.
J Sci Food Agric ; 101(7): 2863-2868, 2021 May.
Artículo en Inglés | MEDLINE | ID: mdl-33151573

RESUMEN

BACKGROUND: Detection of chlorophyll metabolites in milk has recently been suggested to be an indicator of a grass-fed diet fed for cattle. Such a means of detection, however, is complicated when the grazing season is over because cattle can be fed fermented silage ingredients, such as alfalfa and corn silage. During fermentation, chlorophyll compounds and other pigments undergo degradation due to the accumulation of lactic acid and the resultant decline in pH. RESULTS: We monitored degradation of chlorophyll compounds by measuring the fluorescence and absorption spectra of silage extracts. The spectroscopic evidence supports the hypothesis that chlorophylls are converted into fluorescent products, such as pheophytin, and further cleaved into pheophorbide. The degradation starts with dechelation and removal of the magnesium ion to produce pheophytin. Further removal of the phytol chain from pheophytin results in the production of pheophorbide. CONCLUSIONS: The fluorescence intensity of these degradation products is reduced compared to that of the parent molecule. These findings are important in understanding the fluorescent signal in milk when cows consume silage rather than fresh pasture grass. © 2020 Society of Chemical Industry.


Asunto(s)
Bovinos/metabolismo , Clorofila/química , Leche/química , Ensilaje/análisis , Anaerobiosis , Alimentación Animal/análisis , Animales , Clorofila/metabolismo , Digestión , Femenino , Fermentación , Medicago sativa/química , Medicago sativa/metabolismo , Leche/metabolismo , Rumen/química , Rumen/metabolismo , Espectrofotometría Atómica , Zea mays/química , Zea mays/metabolismo
6.
J Agric Food Chem ; 66(9): 2168-2173, 2018 Mar 07.
Artículo en Inglés | MEDLINE | ID: mdl-29356527

RESUMEN

Owing to its high ω-3 fatty acid content, milk from grass-fed dairy cows is becoming increasingly more attractive to consumers. Consequently, it is important to identify the origins of such products and to measure their content, at least relative to some standard. To date, chromatography has been the most extensively used technique. Sample preparation and cost, however, often reduce its widespread applicability. Here, we report the effectiveness of fluorescence spectroscopy for such quantification by measuring the amount of chlorophyll metabolites in the sample. Their content is significantly higher for milk from grass-fed cows compared to milk from grain/silage-fed cows. It is 0.11-0.13 µM in milk samples from grass-fed cows, whereas in milk from cows fed grain/silage rations, the concentration was 0.01-0.04 µM. In various organic milk samples, the chlorophyll metabolite concentration was in the range of 0.07-0.09 µM. In addition, we explored the mechanisms of photodegradation of milk. Riboflavin and chlorophyll metabolites act as photosensitizers in milk for type-I and type-II reactions, respectively. It was also observed that the presence of high levels of chlorophyll metabolites can synergistically degrade riboflavin, contributing to the degradation of milk quality.


Asunto(s)
Alimentación Animal/análisis , Bovinos/metabolismo , Leche/química , Espectrometría de Fluorescencia/métodos , Animales , Clorofila/química , Clorofila/metabolismo , Luz , Leche/efectos de la radiación , Fotólisis , Poaceae/metabolismo , Riboflavina/química , Riboflavina/metabolismo
7.
J Food Prot ; 80(11): 1815-1820, 2017 11.
Artículo en Inglés | MEDLINE | ID: mdl-28981377

RESUMEN

Because some significant outbreaks of human salmonellosis have been traced to contaminated animal feed, the rapid and efficient detection of Salmonella in feed is essential. However, the current U.S. Food and Drug Administration Bacteriological Analytical Manual (BAM) culture method that uses lactose broth as a preenrichment medium has not reliably supported the results of real-time PCR assays for certain foods. We evaluated the BAM culture method and a quantitative real-time PCR (qPCR) assay using two preenrichment media, modified buffered peptone water and lactose broth, to detect Salmonella enterica subsp. enterica serovar Cubana in naturally contaminated chick feed. After 24 h of incubation, the qPCR method was as sensitive as the culture method when modified buffered peptone water was used as the preenrichment medium but less sensitive than culture when lactose broth was used. After 48 h of incubation, detection of Salmonella Cubana by qPCR and by culture in either preenrichment medium was equivalent. We also compared the performance of the traditional serotyping method, which uses pure cultures of Salmonella grown on blood agar, to two molecular serotyping methods. The serotyping method based on whole genome sequencing also requires pure cultures, but the PCR-based molecular serotyping method can be done directly with the enriched culture medium. The PCR-based molecular serotyping method provided simple and rapid detection and identification of Salmonella Cubana. However, whole genome sequencing allows accurate identification of many Salmonella serotypes and highlights variations in the genomes, even in tight genomic clusters. We also compared the genome of the chick feed isolate with 58 Salmonella Cubana strains in GenBank and found that the chick feed isolate was very closely related to an isolate from a foodborne outbreak involving alfalfa sprouts.


Asunto(s)
Alimentación Animal , Salmonella enterica , Alimentación Animal/microbiología , Animales , Técnicas Bacteriológicas , Tampones (Química) , Pollos , Medios de Cultivo , Humanos , Reacción en Cadena de la Polimerasa/métodos , Salmonella enterica/aislamiento & purificación , Serogrupo , Serotipificación , Verduras/microbiología
8.
J Agric Food Chem ; 64(1): 320-5, 2016 Jan 13.
Artículo en Inglés | MEDLINE | ID: mdl-26623498

RESUMEN

Transmissible spongiform encephalopathies (TSE) are progressive, neurodegenerative disorders, of which bovine spongiform encephalopathy (BSE) is of special concern because it is infectious and debilitating to humans. The possibility of using fluorescence spectroscopy to screen for BSE in cattle was explored. Fluorescence spectra from the retinas of experimentally infected BSE-positive cattle with clinical disease were compared with those from both sham-inoculated and non-inoculated BSE-negative cattle. The distinct intensity difference of about 4-10-fold between the spectra of the BSE-positive and the BSE-negative (sham-inoculated and non-inoculated) eyes suggests the basis for a means of developing a rapid, noninvasive examination of BSE in particular and TSEs in general.


Asunto(s)
Encefalopatía Espongiforme Bovina/diagnóstico , Retina/química , Espectrometría de Fluorescencia/métodos , Animales , Bovinos
9.
Blood ; 126(18): 2146-52, 2015 Oct 29.
Artículo en Inglés | MEDLINE | ID: mdl-26353910

RESUMEN

Drug-dependent antibodies (DDAbs) that cause acute thrombocytopenia upon drug exposure are nonreactive in the absence of the drug but bind tightly to a platelet membrane glycoprotein, usually α(IIb)/ß3 integrin (GPIIb/IIIa) when the drug is present. How a drug promotes binding of antibody to its target is unknown and is difficult to study with human DDAbs, which are poly-specific and in limited supply. We addressed this question using quinine-dependent murine monoclonal antibodies (mAbs), which, in vitro and in vivo, closely mimic antibodies that cause thrombocytopenia in patients sensitive to quinine. Using surface plasmon resonance (SPR) analysis, we found that quinine binds with very high affinity (K(D) ≈ 10⁻9 mol/L) to these mAbs at a molar ratio of ≈ 2:1 but does not bind detectably to an irrelevant mAb. Also using SPR analysis, GPIIb/IIIa was found to bind monovalently to immobilized mAb with low affinity in the absence of quinine and with fivefold greater affinity (K(D) ≈ 2.2 × 10⁻6) when quinine was present. Measurements of quinine-dependent binding of intact mAb and fragment antigen-binding (Fab) fragments to platelets showed that affinity is increased 10 000- to 100 000-fold by bivalent interaction between antibody and its target. Together, the findings indicate that the first step in drug-dependent binding of a DDAb is the interaction of the drug with antibody, rather than with antigen, as has been widely thought, where it induces structural changes that enhance the affinity/specificity of antibody for its target epitope. Bivalent binding may be essential for a DDAb to cause thrombocytopenia.


Asunto(s)
Analgésicos no Narcóticos/inmunología , Anticuerpos Monoclonales/inmunología , Plaquetas/inmunología , Complejo GPIIb-IIIa de Glicoproteína Plaquetaria/inmunología , Quinina/inmunología , Animales , Epítopos/inmunología , Humanos , Fragmentos Fab de Inmunoglobulinas/inmunología , Ratones
10.
Genome Announc ; 2(5)2014 Oct 09.
Artículo en Inglés | MEDLINE | ID: mdl-25301652

RESUMEN

We report the draft genome sequences of Streptococcus bovis strain ATCC 33317 (CVM42251) isolated from cow dung and strain JB1 (CVM42252) isolated from a cow rumen in 1977. The strains were sequenced using the Genome Sequencer FLX 454 system. The genome sizes are approximately 2 Mb and 2.2 Mb, respectively.

11.
Genome Announc ; 2(1)2014 Jan 23.
Artículo en Inglés | MEDLINE | ID: mdl-24459266

RESUMEN

We report the draft genomes of Salmonella enterica subsp. enterica serovar Cubana strain CVM42234, isolated from chick feed in 2012, and S. Cubana strain 76814, isolated from swine in 2004. The genome sizes are 4,975,046 and 4,936,251 bp, respectively.

12.
Foodborne Pathog Dis ; 10(8): 684-91, 2013 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-23692074

RESUMEN

The presence and antimicrobial susceptibility of foodborne pathogens and indicator organisms in animal feed are not well understood. In this study, a total of 201 feed ingredient samples (animal byproducts, n=122; plant byproducts, n=79) were collected in 2002 and 2003 from representative rendering plants and the oilseed (or cereal grain) industry across the United States. The occurrence and antimicrobial susceptibility of four bacterial genera (Salmonella, Campylobacter, Escherichia coli, and Enterococcus) were determined. Salmonella isolates were further characterized by serotyping and pulsed-field gel electrophoresis (PFGE). None of the samples yielded Campylobacter or E. coli O157:H7, whereas Salmonella, generic E. coli, and Enterococcus were present in 22.9%, 39.3%, and 86.6% of samples, respectively. A large percentage (47.8%) of Salmonella-positive samples harbored two serovars, and the vast majority (88.4%) of Enterococcus isolates were E. faecium. Animal byproducts had a significantly higher Salmonella contamination rate (34.4%) than plant byproducts (5.1%) (p<0.05). Among 74 Salmonella isolates recovered, 27 serovars and 55 PFGE patterns were identified; all were pan-susceptible to 17 antimicrobials tested. E. coli isolates (n=131) demonstrated similar susceptibility to these antimicrobials except for tetracycline (15.3% resistance), sulfamethoxazole (7.6%), streptomycin (4.6%), ampicillin (3.8%), and nalidixic acid (1.5%). Enterococcus isolates (n=362) were also resistant to five of 17 antimicrobials tested, ranging from 1.1% to penicillin to 14.6% to tetracycline. Resistance rates were generally higher among isolates recovered from animal byproducts. Taken together, our findings suggest that diverse populations of Salmonella, E. coli, and Enterococcus are commonly present in animal feed ingredients, but antimicrobial resistance is not common. Future large-scale studies to monitor these pathogenic and indicator organisms in feed commodities is warranted.


Asunto(s)
Alimentación Animal/microbiología , Campylobacter/aislamiento & purificación , Enterococcus/aislamiento & purificación , Escherichia coli/aislamiento & purificación , Carne/microbiología , Salmonella/aislamiento & purificación , Animales , Campylobacter/efectos de los fármacos , Recuento de Colonia Microbiana , Farmacorresistencia Bacteriana Múltiple/efectos de los fármacos , Electroforesis en Gel de Campo Pulsado , Enterococcus/efectos de los fármacos , Escherichia coli/efectos de los fármacos , Contaminación de Alimentos , Microbiología de Alimentos , Pruebas de Sensibilidad Microbiana , Salmonella/efectos de los fármacos , Serotipificación , Estados Unidos
13.
Photochem Photobiol ; 89(4): 864-8, 2013.
Artículo en Inglés | MEDLINE | ID: mdl-23360345

RESUMEN

Recently, we have proposed that the fluorescence spectra of sheep retina can be well correlated with the presence or absence of scrapie. Scrapie is the most widespread TSE (transmissible spongiform encephalopathy) affecting sheep and goats worldwide. Mice eyes have been previously reported as a model system to study age-related accumulation of lipofuscin, which has been investigated by monitoring the increasing fluorescence with age covering its entire life span. The current work aims at developing mice retina as a convenient model system to diagnose scrapie and other fatal TSE diseases in animals such as sheep and cows. The objective of the research reported here was to determine whether the spectral features are conserved between two different species namely mice and sheep, and whether an appropriate small animal model system could be identified for diagnosis of scrapie based on the fluorescence intensity in retina. The results were consistent with the previous reports on fluorescence studies of healthy and scrapie-infected retina of sheep. The fluorescence from the retinas of scrapie-infected sheep was significantly more intense and showed more heterogeneity than that from the retinas of uninfected mice. Although the structural characteristics of fluorescence spectra of scrapie-infected sheep and mice eyes are slightly different, more importantly, murine retinas reflect the enhancement of fluorescence intensity upon infecting the mice with scrapie, which is consistent with the observations in sheep eyes.


Asunto(s)
Retina/patología , Scrapie/patología , Espectrometría de Fluorescencia , Animales , Femenino , Masculino , Ratones
14.
J Agric Food Chem ; 60(42): 10456-60, 2012 Oct 24.
Artículo en Inglés | MEDLINE | ID: mdl-23002917

RESUMEN

Efflux pumps are vital bacterial components, and research has demonstrated that some plant compounds such as pheophorbide a (php) possess efflux pump inhibitor (EPI) activity. This study determined the quantity of php present in feces as an indicator of EPI activity. Feces were collected from different species of animals fed a variety of feeds. The chlorophyll metabolites php and pyropheophorbide a (pyp) were determined using fluorescense spectroscopy. The average concentrations [µg/g dry matter (DM) feces] of pyp/php in feces were as follows: guinea pig, 180; goat, 150; rabbit, 150; dairy cow, 120; feedlot cattle, 60; rat, <1; pig, <1; chicken, <1. These data indicate that animals consuming "green" diets will excrete feces with concentrations of php/pyp that exceed levels demonstrated to be inhibitory to bacterial efflux pumps (0.5 µg/mL). The natural presence EPIs in the gastrointestinal tract may modulate the activity of microbial efflux pumps and exert selection pressure upon resident microbial populations.


Asunto(s)
Clorofila/análogos & derivados , Heces/química , Espectrometría de Fluorescencia/métodos , Animales , Clorofila/análisis
15.
Blood ; 119(26): 6317-25, 2012 Jun 28.
Artículo en Inglés | MEDLINE | ID: mdl-22490676

RESUMEN

Arginine-glycine-aspartic acid (RGD)-mimetic platelet inhibitors act by occupying the RGD recognition site of α(IIb)/ß(3) integrin (GPIIb/IIIa), thereby preventing the activated integrin from reacting with fibrinogen. Thrombocytopenia is a well-known side effect of treatment with this class of drugs and is caused by Abs, often naturally occurring, that recognize α(IIb)/ß(3) in a complex with the drug being administered. RGD peptide and RGD-mimetic drugs are known to induce epitopes (ligand-induced binding sites [LIBS]) in α(IIb)/ß(3) that are recognized by certain mAbs. It has been speculated, but not shown experimentally, that Abs from patients who develop thrombocytopenia when treated with an RGD-mimetic inhibitor similarly recognize LIBS determinants. We addressed this question by comparing the reactions of patient Abs and LIBS-specific mAbs against α(IIb)/ß(3) in a complex with RGD and RGD-mimetic drugs, and by examining the ability of selected non-LIBS mAbs to block binding of patient Abs to the liganded integrin. Findings made provide evidence that the patient Abs recognize subtle, drug-induced structural changes in the integrin head region that are clustered about the RGD recognition site. The target epitopes differ from classic LIBS determinants, however, both in their location and by virtue of being largely drug-specific.


Asunto(s)
Anticuerpos/efectos adversos , Anticuerpos/metabolismo , Complejo GPIIb-IIIa de Glicoproteína Plaquetaria/inmunología , Complejo GPIIb-IIIa de Glicoproteína Plaquetaria/metabolismo , Trombocitopenia/etiología , Animales , Anticuerpos/inmunología , Células CHO , Cricetinae , Cricetulus , Eptifibatida , Humanos , Ligandos , Ratones , Ratones Endogámicos BALB C , Modelos Biológicos , Oligopéptidos/química , Oligopéptidos/inmunología , Péptidos/uso terapéutico , Inhibidores de Agregación Plaquetaria/química , Inhibidores de Agregación Plaquetaria/uso terapéutico , Complejo GPIIb-IIIa de Glicoproteína Plaquetaria/química , Complejo GPIIb-IIIa de Glicoproteína Plaquetaria/genética , Unión Proteica/inmunología , Conformación Proteica , Ratas , Especificidad por Sustrato , Trombocitopenia/tratamiento farmacológico , Trombocitopenia/inmunología , Tirofibán , Tirosina/análogos & derivados , Tirosina/uso terapéutico
16.
Sci Transl Med ; 4(125): 125ra32, 2012 Mar 14.
Artículo en Inglés | MEDLINE | ID: mdl-22422993

RESUMEN

An integrin found on platelets, α(IIb)ß(3) mediates platelet aggregation, and α(IIb)ß(3) antagonists are effective antithrombotic agents in the clinic. Ligands bind to integrins in part by coordinating a magnesium ion (Mg(2+)) located in the ß subunit metal ion-dependent adhesion site (MIDAS). Drugs patterned on the integrin ligand sequence Arg-Gly-Asp have a basic moiety that binds the α(IIb) subunit and a carboxyl group that coordinates the MIDAS Mg(2+) in the ß(3) subunits. They induce conformational changes in the ß(3) subunit that may have negative consequences such as exposing previously hidden epitopes and inducing the active conformation of the receptor. We recently reported an inhibitor of α(IIb)ß(3) (RUC-1) that binds exclusively to the α(IIb) subunit; here, we report the structure-based design and synthesis of RUC-2, a RUC-1 derivative with a ~100-fold higher affinity. RUC-2 does not induce major conformational changes in ß(3) as judged by monoclonal antibody binding, light scattering, gel chromatography, electron microscopy, and a receptor priming assay. X-ray crystallography of the RUC-2-α(IIb)ß(3) headpiece complex in 1 mM calcium ion (Ca(2+))/5 mM Mg(2+) at 2.6 Å revealed that RUC-2 binds to α(IIb) the way RUC-1 does, but in addition, it binds to the ß(3) MIDAS residue glutamic acid 220, thus displacing Mg(2+) from the MIDAS. When the Mg(2+) concentration was increased to 20 mM, however, Mg(2+) was identified in the MIDAS and RUC-2 was absent. RUC-2's ability to inhibit ligand binding and platelet aggregation was diminished by increasing the Mg(2+) concentration. Thus, RUC-2 inhibits ligand binding by a mechanism different from that of all other α(IIb)ß(3) antagonists and may offer advantages as a therapeutic agent.


Asunto(s)
Magnesio/metabolismo , Complejo GPIIb-IIIa de Glicoproteína Plaquetaria/metabolismo , Animales , Sitios de Unión , Plaquetas/efectos de los fármacos , Plaquetas/metabolismo , Adhesión Celular/fisiología , Colágeno/metabolismo , Cristalografía por Rayos X , Fibrinógeno/metabolismo , Humanos , Ratones , Microscopía Electrónica , Oligopéptidos , Agregación Plaquetaria/efectos de los fármacos , Inhibidores de Agregación Plaquetaria/metabolismo , Inhibidores de Agregación Plaquetaria/farmacología , Unión Proteica/efectos de los fármacos , Unión Proteica/fisiología , Ratas , Vitronectina/metabolismo
17.
Vet Microbiol ; 151(3-4): 301-6, 2011 Aug 05.
Artículo en Inglés | MEDLINE | ID: mdl-21481550

RESUMEN

Recent studies have identified a phenomenon in which ciliated protozoa engulf Salmonella and the intra-protozoal environment hyperactivates virulence gene expression and provides a venue for conjugal transfer of antibiotic resistance plasmids. The former observation is relegated to Salmonella bearing the SGI1 multiresistance integron while the latter phenomenon appears to be a more generalized event for recipient Salmonella. Our previous studies have assessed virulence gene hyperexpression only with protozoa from the bovine rumen while conjugal transfer has been demonstrated in rumen protozoa from cattle and goats. The present study examined virulence gene hyperexpression for Salmonella exposed to rumen protozoa obtained from cattle, sheep, goats, or two African ruminants (giraffe and bongo). Conjugal transfer was also assessed in these protozoa using Salmonella as the recipient. Virulence gene hyperexpression was only observed following exposure to the rumen protozoa from cattle and sheep while elevated virulence was also observed in these animals. Conjugal transfer events were, however, observed in all protozoa evaluated. It therefore appears that the protozoa-based hypervirulence is not universal to all ruminants while conjugal transfer is more ubiquitous.


Asunto(s)
Conjugación Genética , Rumen/microbiología , Rumen/parasitología , Rumiantes/microbiología , Rumiantes/parasitología , Salmonelosis Animal/parasitología , Salmonella/genética , Animales , Animales Salvajes/microbiología , Animales Salvajes/parasitología , Bovinos/microbiología , Bovinos/parasitología , Cilióforos/aislamiento & purificación , Regulación Bacteriana de la Expresión Génica , Genes Bacterianos , Cabras/microbiología , Cabras/parasitología , Integrones/genética , Plásmidos , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Salmonella/patogenicidad , Salmonelosis Animal/microbiología , Ovinos/microbiología , Ovinos/parasitología , Virulencia/genética
18.
Surg Radiol Anat ; 33(1): 53-7, 2011 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-20623121

RESUMEN

The neck is an area that lends itself to anatomical geometry, such as triangles. Many triangles of the neck have been described, and some are well known; however, some have been nearly forgotten. The purposes of this study were to review prior literature on the forgotten triangles of the neck, to measure and document the occurrence of these triangles and the structures found within them, and to discuss their surgical significance and relationships to the standard triangles of the neck. This study was carried out on 17 (34 sides) formaldehyde-fixed adult human cadavers (5 males and 12 females), the mean age being 78 years. All triangles were found within the submandibular triangle of the anterior cervical triangle. Beclard's triangle was present on 28 sides and absent on 6 sides because the posterior belly of the digastric muscle did not touch the hyoid bone and because the posterior belly of the digastric muscle was directly over the greater cornu of the hyoid bone. Lesser's triangle was present on 30 sides and absent on 4 sides because the hypoglossal nerve remained inferior to the digastric muscle. Pirogoff's triangle was also present on 30 sides and absent on 4 sides, again because the hypoglossal nerve remained inferior to the digastric muscle. No statistically significant differences were found between the measurements of each leg of any of the three triangles and gender, or between the measurement of each leg of any of the three triangles and right versus left sides. Beclard's triangle, when present, was found to contain both the hypoglossal nerve and the lingual artery. Pirogoff's triangle always contained the lingual artery, and Lesser's triangle contained no major structures, but the lingual artery was deep to its floor. Knowledge of these triangles, their contents, and relationships to other structures of the neck could be important for cranio-facial, oral, general, and neurosurgeons who operate within the neck.


Asunto(s)
Disección del Cuello , Cuello/anatomía & histología , Anciano , Femenino , Humanos , Masculino
19.
Anal Chem ; 82(10): 4097-101, 2010 May 15.
Artículo en Inglés | MEDLINE | ID: mdl-20411920

RESUMEN

The feasibility of exploiting fluorescence spectra of the eye for diagnosis of transmissible spongiform encephalopathies (TSEs) was examined. Retinas from scrapie-positive sheep were compared with scrapie-negative sheep using fluorescence spectroscopy, and distinct differences in the fluorescence intensity and spectroscopic signatures were observed. The characteristic fluorescent signatures are thought to be the result of an accumulation of lipofuscin in the retina. It appears that the eye, in particular the retina, is a useful tissue for noninvasive examination of some neurological pathologies such as scrapie. The development of procedures based on examinations of the eye that permit the detection of neurological disorders in animals holds great promise.


Asunto(s)
Encefalopatía Espongiforme Bovina/diagnóstico , Enfermedades por Prión/diagnóstico , Retina/patología , Espectrometría de Fluorescencia/métodos , Animales , Encéfalo/patología , Bovinos , Retina/fisiopatología , Scrapie/diagnóstico
20.
Photochem Photobiol ; 85(6): 1322-6, 2009.
Artículo en Inglés | MEDLINE | ID: mdl-19656327

RESUMEN

We describe a comparison of the fluorescence spectra of bovine tissues with murine tissues in order to determine whether spectral features are conserved and whether an appropriate and practical laboratory small animal model system could be identified to be used for investigation of tissue- and age-related fluorescence signal patterns. Recently it has been shown that spectral signatures of lipofuscin have enabled the detection of bovine central nervous system (CNS) tissue in meat products with high sensitivity (Schönenbrücher, H., Adhikary, R., Mukherjee, P., Casey, T.A., Rasmussen, M.A., Maistrovich, F.D., Hamir, A.N., Kehrli, M.J., Richt, J., Petrich, J.W. [2008] J Agric Food Chem56, 6220-6226). We report that brain and spinal cord of mice provide fluorescence spectra similar to those of bovine brain and spinal cord. It is concluded that murine CNS tissue is an appropriate model system for bovine CNS tissue for the development of fluorometric CNS detection assays.


Asunto(s)
Sistema Nervioso Central/anatomía & histología , Sistema Nervioso Central/química , Espectrometría de Fluorescencia , Animales , Bovinos , Contaminación de Alimentos , Ratones , Modelos Animales
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