RESUMEN
Laboratory rats are generally fed ad libitum, although this method is associated with obesity and an increased frequency of spontaneous tumours. It has been challenging looking for ways to limit feed consumption in group-housed rats without any setbacks to animal welfare and scientific results. The diet board, as a method of dietary restriction, was used in the present study. Diet board feeding allows group housing and should result in enhanced welfare compared with traditional methods of dietary restriction. With respect to animal model robustness and translatability of results it is important that the feeding regime does not affect diurnal rhythmicity of biological parameters. In the present study the effects of diet board feeding on diurnal rhythms of blood glucose, serum ghrelin, faecal immunoglobulin A (IgA) and faecal corticosterone were assessed. The diet board did not alter diurnal rhythms, and adds weight to the use of this method for dietary restriction which should benefit animal health and the validity of scientific results generated from the animals.
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Glucemia/metabolismo , Ritmo Circadiano , Corticosterona/metabolismo , Privación de Alimentos , Ghrelina/sangre , Inmunoglobulina A/metabolismo , Ratas/fisiología , Bienestar del Animal , Animales , Dieta , Heces/química , MasculinoRESUMEN
Cnidarians are the oldest extant lineage of venomous animals. Despite their simple anatomy, they are capable of subduing or repelling prey and predator species that are far more complex and recently evolved. Utilizing specialized penetrating nematocysts, cnidarians inject the nematocyst content or "venom" that initiates toxic and immunological reactions in the envenomated organism. These venoms contain enzymes, potent pore forming toxins, and neurotoxins. Enzymes include lipolytic and proteolytic proteins that catabolize prey tissues. Cnidarian pore forming toxins self-assemble to form robust membrane pores that can cause cell death via osmotic lysis. Neurotoxins exhibit rapid ion channel specific activities. In addition, certain cnidarian venoms contain or induce the release of host vasodilatory biogenic amines such as serotonin, histamine, bunodosine and caissarone accelerating the pathogenic effects of other venom enzymes and porins. The cnidarian attacking/defending mechanism is fast and efficient, and massive envenomation of humans may result in death, in some cases within a few minutes to an hour after sting. The complexity of venom components represents a unique therapeutic challenge and probably reflects the ancient evolutionary history of the cnidarian venom system. Thus, they are invaluable as a therapeutic target for sting treatment or as lead compounds for drug design.
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Venenos de Cnidarios , Animales , Cnidarios/genética , Cnidarios/fisiología , Venenos de Cnidarios/química , Venenos de Cnidarios/toxicidad , Descubrimiento de Drogas , Humanos , FilogeniaRESUMEN
Animal research has made major contributions to the health and welfare of humans and domestic animals. Immunization, first developed against rabies and anthrax by Pasteur using dogs, sheep, and rabbits, is now used to control many infectious diseases. The first drug, Salvarsan, was developed by Ehrlich using rabbits infected with the organism causing syphilis. This was the forerunner of the many drugs developed by the pharmaceutical industry today. The discovery of vitamins using rats has almost eliminated diseases such as scurvy and rickets; and hormones, such as insulin discovered following work in dogs, have helped to control many metabolic diseases. These and many other advances have enabled physicians to treat a wide range of human diseases. But many diseases continue to cause suffering.
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Experimentación Animal , Proyectos de Investigación , Animales , HumanosRESUMEN
If the scientist needs to contact the animal facility after any study to inquire about husbandry details, this represents a lost opportunity, which can ultimately interfere with the study results and their interpretation. There is a clear tendency for authors to describe methodological procedures down to the smallest detail, but at the same time to provide minimal information on animals and their husbandry. Controlling all major variables as far as possible is the key issue when establishing an experimental design. The other common mechanism affecting study results is a change in the variation. Factors causing bias or variation changes are also detectable within husbandry. Our lives and the lives of animals are governed by cycles: the seasons, the reproductive cycle, the weekend-working days, the cage change/room sanitation cycle, and the diurnal rhythm. Some of these may be attributable to routine husbandry, and the rest are cycles, which may be affected by husbandry procedures. Other issues to be considered are consequences of in-house transport, restrictions caused by caging, randomization of cage location, the physical environment inside the cage, the acoustic environment audible to animals, olfactory environment, materials in the cage, cage complexity, feeding regimens, kinship, and humans. Laboratory animal husbandry issues are an integral but underappreciated part of investigators' experimental design, which if ignored can cause major interference with the results. All researchers should familiarize themselves with the current routine animal care of the facility serving them, including their capabilities for the monitoring of biological and physicochemical environment.
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Experimentación Animal , Crianza de Animales Domésticos , Proyectos de Investigación , Bienestar del Animal , Animales , Ritmo Circadiano , Vivienda para AnimalesRESUMEN
Cellular distribution of group XIIA phospholipase A2 (GXIIA PLA2) was studied in human digestive organs by immunohistochemistry. GXIIA PLA2 protein was detected in epithelial cells of normal gastrointestinal tract, gallbladder and pancreatic acinar cells. The GXIIA PLA2 protein was evenly distributed in the cytoplasm in contrast to secretory granular distribution of GIB PLA2 and GIIA PLA2 in pancreatic acinar cells and small intestinal Paneth cells respectively. Epithelial cells of intestinal glands in Crohn's disease and ulcerative colitis expressed abundant GXIIA PLA2 , whereas inflammatory cells were devoid of the enzyme protein. Tumour cells in colonic adenomas and carcinomas and pancreatic ductogenic carcinomas expressed GXIIA PLA2 protein at varying intensity levels. The putative functions of GXIIA PLA2 remain to be investigated and its role in healthy and diseased digestive organs can only be speculated on at present.
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Regulación Enzimológica de la Expresión Génica , Fosfolipasas A2/metabolismo , Adulto , Anciano , Anciano de 80 o más Años , Secuencia de Aminoácidos , Línea Celular Tumoral , Clonación Molecular , Colitis Ulcerosa/genética , Colitis Ulcerosa/patología , Enfermedad de Crohn/genética , Enfermedad de Crohn/patología , Células Epiteliales/metabolismo , Femenino , Vesícula Biliar/citología , Vesícula Biliar/enzimología , Tracto Gastrointestinal/citología , Tracto Gastrointestinal/enzimología , Humanos , Inmunohistoquímica , Mucosa Intestinal/citología , Mucosa Intestinal/patología , Masculino , Persona de Mediana Edad , Datos de Secuencia Molecular , Páncreas/citología , Páncreas/enzimología , Fosfolipasas A2/genética , ARN Mensajero/genética , ARN Mensajero/metabolismoRESUMEN
Sequence homologues of the bacterium Streptomyces violaceoruber and sea anemone Nematostella vectensis PLA2 pfam09056 members were identified in several bacteria, fungi and metazoans illustrating the evolution of this PLA2 sub-family. Comparison of their molecular structures revealed that bacteria and fungi members are part of the GXIV of PLA2s while metazoan representatives are similar with GIX PLA2 of the marine snail Conus magus. Members of GXIV and GIX PLA2s show modest overall sequence similarity (21-35%) but considerable motif conservation within the putative Ca(2+)-binding, catalytic sites and cysteine residue positions which are essential for enzyme function. GXIV PLA2s of bacteria and fungi typically contain four cysteine residues composing two intramolecular disulphide bonds. GIX PLA2 homologues were identified in cnidarians and molluscs and in a single tunicate but appear to be absent from other metazoan genomes. The mature GIX PLA2 deduced peptides contain up to ten cysteine residues capable of forming five putative disulphide bonds. Three disulphide bonds were identified in GIX PLA2s, two of which correspond to those localized in GXIV PLA2s. Phylogenetic analysis demonstrates that metazoan GIX PLA2s cluster separate from the bacterial and fungal GXIV PLA2s and both pfam09056 members form a group separate from the prokaryote and eukaryote GXIIA PLA2 pfam06951. Duplicate PLA2 pfam09056 genes were identified in the genomes of sea anemone N. vectensis and oyster Crassostrea gigas suggest that members of this family evolved via species-specific duplication events. These observations indicate that the newly identified metazoan pfam09056 members may be classified as GIX PLA2s and support the idea of the common evolutionary origin of GXIV and GIX PLA2 pfam09056 members, which emerged early in bacteria and were maintained in the genomes of fungi and selected extant metazoan taxa.
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Bacterias/genética , Evolución Molecular , Hongos/genética , Invertebrados/clasificación , Invertebrados/genética , Fosfolipasas A2 Secretoras/genética , Filogenia , Animales , Bacterias/química , Bacterias/enzimología , Secuencia Conservada , Hongos/química , Hongos/enzimología , Invertebrados/enzimología , Fosfolipasas A2 Secretoras/química , Alineación de SecuenciaRESUMEN
The 3Rs--replacement, reduction, and refinement--are aimed at minimizing the welfare costs to animals used in research. Some neuroscientists fear that implementing the 3Rs will prohibit essential studies. Others view them as fundamental ethical principles that improve the quality of research. A regulatory system that integrates science and welfare is most likely to deliver public confidence.
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Alternativas a las Pruebas en Animales , Investigación Biomédica , Neurociencias , Animales , Investigación Biomédica/ética , Investigación Biomédica/métodos , Humanos , Neurociencias/éticaRESUMEN
Vertebrate group XII phospholipases A(2) (GXII PLA(2), conserved domain pfam06951) are proteins with unique structural and functional features within the secreted PLA(2) family. In humans, two genes (GXIIA PLA(2) and GXIIB PLA(2)) have been characterised. GXIIA PLA(2) is enzymatically active whereas GXIIB PLA(2) is devoid of catalytic activity. Recently, putative homologues of the vertebrate GXII PLA(2)s were described in non-vertebrates. In the current study a total of 170 GXII PLA(2) sequences were identified in vertebrates, invertebrates, non-metazoan eukaryotes, fungi and bacteria. GXIIB PLA(2) was found only in vertebrates and the searches failed to identify putative GXII PLA(2) homologues in Archaea. Comparisons of the predicted functional domains of GXII PLA(2)s revealed considerable structural identity within the Ca(2+)-binding and the catalytic sites among the various organisms suggesting that functional conservation may have been retained across evolution. The preservation of GXII PLA(2) family members from bacteria to human indicates that they have emerged early in evolution and evolved via gene/genome duplication events prior to Eubacteria. Gene duplicates were identified in some invertebrate taxa suggesting that species-specific duplications occurred. The analysis of the GXII PLA(2) homologue genome environment revealed that gene synteny and gene order are preserved in vertebrates. Conservation of GXII PLA(2)s indicates that important functional roles involved in species survival and were maintained across evolution and may be dependent on or independent of the enzyme's phospholipolytic activity.
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Bacterias/enzimología , Secuencia Conservada , Fosfolipasas A2/metabolismo , Animales , HumanosRESUMEN
Secreted phospholipases A(2) (sPLA(2) s) are lipolytic enzymes present in organisms ranging from prokaryotes to eukaryotes but their origin and emergence are poorly understood. We identified and compared the conserved domains of 333 sPLA(2) s and proposed a model for their evolution. The conserved domains were grouped into seven categories according to the in silico annotated conserved domain collections of 'cd00618: PLA(2) _like' and 'pfam00068: Phospholip_A2_1'. PLA(2) s containing the conserved domain cd04706 (plant-specific PLA(2) ) are present in bacteria and plants. Metazoan PLA(2) s of the group (G) I/II/V/X PLA(2) collection exclusively contain the conserved domain cd00125. GIII PLA(2) s of both vertebrates and invertebrates contain the conserved domain cd04704 (bee venom-like PLA(2) ), and mammalian GIII PLA(2) s also contain the conserved domain cd04705 (similar to human GIII PLA(2) ). The sPLA(2) s of bacteria, fungi and marine invertebrates contain the conserved domain pfam09056 (prokaryotic PLA(2) ) that is the only conserved domain identified in fungal sPLA(2) s. Pfam06951 (GXII PLA(2) ) is present in bacteria and is widely distributed in eukaryotes. All conserved domains were present across mammalian sPLA(2) s, with the exception of cd04706 and pfam09056. Notably, no sPLA(2) s were found in Archaea. Phylogenetic analysis of sPLA(2) conserved domains reveals that two main clades, the cd- and the pfam-collection, exist, and that they have evolved via gene-duplication and gene-deletion events. These observations are consistent with the hypothesis that sPLA(2) s in eukaryotes shared common origins with two types of bacterial sPLA(2) s, and their persistence during evolution may be related to their role in phospholipid metabolism, which is fundamental for survival.
Asunto(s)
Secuencia Conservada/genética , Evolución Molecular , Fosfolipasas A2/genética , Alineación de Secuencia/métodos , Secuencia de Aminoácidos , Animales , Bacterias/enzimología , Bacterias/genética , Sitios de Unión/genética , Bases de Datos Genéticas , Células Eucariotas/enzimología , Células Eucariotas/metabolismo , Humanos , Datos de Secuencia Molecular , Fosfolipasas A2/clasificación , Fosfolipasas A2/metabolismo , Filogenia , Plantas/enzimología , Plantas/genética , Células Procariotas/enzimología , Células Procariotas/metabolismo , Homología de Secuencia de Aminoácido , Especificidad de la EspecieRESUMEN
This paper reviews the animal welfare challenges associated with the use of minipigs in toxicology testing, and compares these to published knowledge on the other widely used non-rodent species (dogs and non-human primates). Welfare challenges arise from housing and management of populations under laboratory conditions, and from the procedures carried out for product evaluation. Welfare assessment requires a multidisciplinary approach: cardiovascular parameters, adrenocortical hormones and behaviour are well known parameters. However, reliable non-invasive methods to assess welfare and species-specific benchmarks need further development in minipigs. Husbandry of minipigs (housing, diet, and socialisation needs) to promote good welfare is described in the revised Appendix A of the European Convention (ETS 123). This has been supplemented by knowledge of species biology and expert opinion from experienced minipig users. Challenges when using minipigs in toxicity testing have been reviewed in detail. Although deeper location of the peripheral blood vessels makes blood sampling more challenging, samples can be taken with minimal distress when staff members are well trained. Temporary and chronic vascular catheters can also be used for frequent sampling, and are likely to improve the welfare of the animals. Available training courses with a focus on stress free handling and dosing, as well as surgical placement of temporary and chronic vascular catheters, should be utilised to improve welfare during these procedures. Humane endpoints have been described, mainly based on current industry practices, but further scientific investigations are required. From an animal welfare perspective there are no basic restrictions to using minipigs in toxicity testing that are unique to this species. We conclude that it is easier to keep minipigs to a good standard of welfare under laboratory conditions than it is for dogs or non-human primates, since minipigs are not athletic (like dogs) or arboreal (like non-human primates).
Asunto(s)
Bienestar del Animal , Porcinos Enanos , Pruebas de Toxicidad , Crianza de Animales Domésticos , Animales , Animales de Laboratorio/fisiología , Conducta Animal , Unión Europea , Femenino , Regulación Gubernamental , Humanos , Masculino , Proyectos de Investigación , Porcinos , Porcinos Enanos/fisiología , Pruebas de Toxicidad/normasRESUMEN
Peroxiredoxins (Prx) are enzymes that catalyze the reduction of hydrogen peroxide and alkyl hydroperoxides. Prxs are ubiquitous enzymes with representatives found in Bacteria, Archaea and Eukarya. Many 1-cysteine peroxiredoxins (1-CysPrx) are dual-function enzyme with both peroxidase and acidic Ca(2+)-independent phospholipase A(2) (aiPLA(2)) activities. The functions proposed for 1-CysPrx/aiPLA(2) include the protection of cell membrane phospholipids against oxidative damage (peroxidation) and the metabolism (hydrolysis) of phospholipids, such as those of lung surfactant. The peroxidase active site motif PVCTTE of 1-CysPrx contains the conserved catalytic cysteine residue, and the esterase (lipase) motif GXSXG of the enzyme contains the conserved catalytic serine residue. In addition to the classic lipase motif GXSXG, various 1-CysPrx/aiPLA(2)s have closely related variant putative lipase motifs containing the catalytic serine residue. The PLA(2) moieties are prevalent and highly homologous in vertebrate and bacterial 1-CysPrx/aiPLA(2)s that is consistent with a high degree evolutional conservation of the enzyme.
Asunto(s)
Cisteína/metabolismo , Peroxidasas/metabolismo , Peroxirredoxinas/metabolismo , Fosfolipasas A2 Calcio-Independiente/metabolismo , Animales , Catálisis , Cisteína/química , Humanos , Estrés Oxidativo , Fosfolipasas A2 Citosólicas/metabolismo , FilogeniaRESUMEN
Phospholipases A(2) (PLA(2)) are enzymes which cleave the sn-2 ester bond in membrane phospholipids to release free fatty acids and lysophospholipids. The present study aimed to elucidate the expression profile of multiple secretory phospholipase A(2) (sPLA(2)) isoforms in the normal rat CNS with focus on sPLA(2)-IIA in the brainstem and spinal cord. Quantitative RT-PCR analysis showed that sPLA(2)-IB expression was low throughout the CNS, sPLA(2)-IIA expression was high in the brainstem and spinal cord, sPLA(2)-IIC expression was high in the cerebral neocortex, hippocampus and thalamus/hypothalamus, sPLA(2)-V expression was high in the olfactory bulb and cerebellum, and sPLA(2)-X was expressed at very low levels in the normal CNS. Of the isoforms, sPLA(2)-IIA mRNA expression was highest in the brainstem and spinal cord suggesting that this could be the most relevant isoform in the ascending pain pathway. Western blot analysis showed high level of sPLA(2)-IIA expression in the brainstem and cervical, thoracic and lumbar spinal segments but low level of expression in other parts of the brain. sPLA(2)-IIA was localized by immunohistochemistry to the spinal trigeminal and facial motor nuclei and dorsal- and ventral-horns of the spinal cord. The enzyme was found on the endoplasmic reticulum of neuronal cell bodies and small diameter dendrites or dendritic spines at electron microscopy. The expression of sPLA(2)-IIA in the dorsal horn and spinal trigeminal nucleus is consistent with previous results which showed an important role of CNS sPLA(2) in nociceptive transmission.
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Tronco Encefálico/enzimología , Regulación Enzimológica de la Expresión Génica/fisiología , Fosfolipasas A2 Secretoras/genética , Médula Espinal/enzimología , Animales , Mapeo Encefálico , Tronco Encefálico/citología , Dendritas/metabolismo , Dendritas/ultraestructura , Retículo Endoplásmico/metabolismo , Retículo Endoplásmico/ultraestructura , Perfilación de la Expresión Génica , Inmunohistoquímica , Isoenzimas/genética , Masculino , Microscopía Electrónica de Transmisión , Neuronas/metabolismo , Neuronas/ultraestructura , Nociceptores/citología , Nociceptores/enzimología , Fosfolipasas A2 Secretoras/metabolismo , Células del Asta Posterior/citología , Células del Asta Posterior/enzimología , Prosencéfalo/citología , Prosencéfalo/enzimología , ARN Mensajero/análisis , ARN Mensajero/metabolismo , Ratas , Ratas Wistar , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Médula Espinal/citología , Núcleo Caudal del Trigémino/citología , Núcleo Caudal del Trigémino/enzimologíaRESUMEN
Throughout evolution, numerous proteins have been convergently recruited into the venoms of various animals, including centipedes, cephalopods, cone snails, fish, insects (several independent venom systems), platypus, scorpions, shrews, spiders, toxicoferan reptiles (lizards and snakes), and sea anemones. The protein scaffolds utilized convergently have included AVIT/colipase/prokineticin, CAP, chitinase, cystatin, defensins, hyaluronidase, Kunitz, lectin, lipocalin, natriuretic peptide, peptidase S1, phospholipase A(2), sphingomyelinase D, and SPRY. Many of these same venom protein types have also been convergently recruited for use in the hematophagous gland secretions of invertebrates (e.g., fleas, leeches, kissing bugs, mosquitoes, and ticks) and vertebrates (e.g., vampire bats). Here, we discuss a number of overarching structural, functional, and evolutionary generalities of the protein families from which these toxins have been frequently recruited and propose a revised and expanded working definition for venom. Given the large number of striking similarities between the protein compositions of conventional venoms and hematophagous secretions, we argue that the latter should also fall under the same definition.
Asunto(s)
Proteínas/genética , Toxicogenética , Ponzoñas/genética , Ponzoñas/toxicidad , Adaptación Biológica , Animales , Genoma , Humanos , FilogeniaRESUMEN
Among mammalian secreted phospholipases A2 (sPLA(2)s), the group X enzyme has the most potent hydrolyzing capacity toward phosphatidylcholine, the major phospholipid of cell membrane and lipoproteins. This enzyme has recently been implicated in chronic inflammatory diseases such as atherosclerosis and asthma and may also play a role in colon tumorigenesis. We show here that group X sPLA(2) [mouse (m)GX] is one of the most highly expressed PLA(2) in the mouse colon and that recombinant mouse and human enzymes stimulate proliferation and mitogen-activated protein kinase activation of various colon cell lines, including Colon-26 cancer cells. Among various recombinant sPLA(2)s, mGX is the most potent enzyme to stimulate cell proliferation. Based on the use of sPLA(2) inhibitors, catalytic site mutants, and small interfering RNA silencing of cytosolic PLA(2)alpha and M-type sPLA(2) receptor, we demonstrate that mGX promotes cell proliferation independently of the receptor and via its intrinsic catalytic activity and production of free arachidonic acid and lysophospholipids, which are mitogenic by themselves. mGX can also elicit the production of large amounts of prostaglandin E2 and other eicosanoids from Colon-26 cells, but these lipid mediators do not play a role in mGX-induced cell proliferation because inhibitors of cyclooxygenases and lipoxygenases do not prevent sPLA(2) mitogenic effects. Together, our results indicate that group X sPLA(2) may play an important role in colon tumorigenesis by promoting cancer cell proliferation and releasing various lipid mediators involved in other key events in cancer progression.
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Proliferación Celular , Neoplasias del Colon/patología , Lípidos/biosíntesis , Fosfolipasas A2/farmacología , Animales , Secuencia de Bases , Biocatálisis , Línea Celular Tumoral , Neoplasias del Colon/metabolismo , Humanos , Hibridación in Situ , Ratones , ARN Interferente Pequeño , Proteínas Recombinantes/farmacología , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
Cage change and gavage are routine procedures in animal facilities, yet little is known about whether housing modifications change responses to these procedures. Telemetric activity and cardiovascular parameters were assessed in this experiment. BN and F344 male rats were housed in open or individually ventilated cages, each containing 3 rats, 1 of which had a transponder. A crossover design was used, in which 2 groups were given dividers made of 2 intersecting boards (1 form contained holes loaded with food pellets; the other did not) and 1 group was given a rectangular tunnel. On day 8 of each 2-wk period, the cages were changed; on day 11, rats were gavaged. The parameters were evaluated for the first hour and for the following 17 h. Baseline values for each rat were subtracted from the corresponding response values. The presence of objects did not affect the responses of F344 rats to cage changing or gavage. In BN rats with IVCs, the presence of the plain divider modified the response to both procedures. Responses to procedures appeared to be dependent on both the strain and the cage object, thus complicating the establishment of valid general recommendations.
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Crianza de Animales Domésticos , Conducta Animal , Privación de Alimentos/fisiología , Hemodinámica/fisiología , Vivienda para Animales , Intubación Gastrointestinal/efectos adversos , Animales , Masculino , Actividad Motora/fisiología , Ratas , Ratas Endogámicas BN , Ratas Endogámicas F344 , TelemetríaRESUMEN
Group IIA Phospholipase A(2) (PLA2-IIA), a key enzyme in arachidonic acid and eicosanoid metabolism, participates in a variety of inflammatory processes but possibly also plays a role in tumor progression in vivo. Our aim was to determine the mRNA and protein expression of PLA2-IIA during prostate cancer progression in localized and metastatic prostate tumors. We evaluated the prognostic significance of PLA2-IIA expression in biochemical recurrence, clinical recurrence and disease-specific survival after surgical treatment. The expression of PLA2-IIA was examined by immunohistochemistry and chromogenic in situ hybridization in tissue microarrays of radical prostatectomy specimens and advanced/metastatic carcinomas. The expression data were analyzed in conjunction with clinical follow-up information and clinicopathological variables. The mRNA and protein expression of PLA2-IIA was significantly increased in Gleason pattern grade 2-4 carcinomas compared with benign prostate (p-values 0.042-0.001). In metastases, the expression was significantly lower than in local cancers (p=0.001). The PLA2-IIA expression correlated positively with Ki-67 and alpha-methylacyl CoA racemase (AMACR) expression. The prognostic evaluation revealed decreased PLA2-IIA protein expression among patients who had died of prostate cancer. In conclusion, PLA2-IIA expression is increased in carcinoma when compared with benign prostate. However, metastatic carcinoma showed decreased expression of PLA2-IIA when compared with primary carcinomas. PLA2-IIA may serve as a marker for highly proliferating, possibly poorly differentiated prostate carcinomas. The protein expression of PLA2-IIA may be diminished in patients who consequently die of prostate cancer.
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Biomarcadores de Tumor/biosíntesis , Carcinoma/diagnóstico , Carcinoma/mortalidad , Fosfolipasas A2 Grupo II/biosíntesis , Neoplasias de la Próstata/diagnóstico , Neoplasias de la Próstata/mortalidad , Anciano , Biomarcadores de Tumor/genética , Carcinoma/secundario , Progresión de la Enfermedad , Fosfolipasas A2 Grupo II/genética , Humanos , Inmunohistoquímica , Hibridación in Situ , Antígeno Ki-67/metabolismo , Masculino , Persona de Mediana Edad , Metástasis de la Neoplasia , Pronóstico , Próstata/enzimología , Próstata/cirugía , Prostatectomía , Neoplasias de la Próstata/patología , ARN Mensajero/genética , Racemasas y Epimerasas/metabolismo , Estudios RetrospectivosRESUMEN
Antibacterial properties of secreted phospholipases A2 (PLA2) have emerged gradually. Group (G) IIA PLA2 is the most potent among mammalian secreted (s) PLA2s against Gram-positive bacteria, but additional antibacterial compounds, e.g. the bactericidal/permeability-increasing protein, are needed to kill Gram-negative bacteria. The mechanisms of binding to the bacterial surface and the killing of bacteria by sPLA2s are based on the positive charge of the PLA2 protein and its phospholipolytic enzymatic activity, respectively. The concentration of GIIA PLA2 is highly elevated in serum of patients with bacterial sepsis, and overexpression of GIIA PLA(2) protects transgenic mice against experimental Gram-positive infection. The synthesis and secretion of GIIA PLA2 are stimulated by the cytokines TNF-alpha, IL-1 and IL-6. Secreted PLA2s may be potentially useful new endogenous antibiotics to combat infections including those caused by antibiotic-resistant bacteria such as methicillin-resistant staphylococci and vancomysin-resistant enterococci.
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Antibacterianos/metabolismo , Antibacterianos/farmacología , Fosfolipasas A2 Secretoras/metabolismo , Fosfolipasas A2 Secretoras/farmacología , Animales , Antibacterianos/clasificación , Infecciones Bacterianas/tratamiento farmacológico , Infecciones Bacterianas/enzimología , Farmacorresistencia Bacteriana/efectos de los fármacos , Humanos , Inflamación/tratamiento farmacológico , Inflamación/enzimología , Fosfolipasas A2 Secretoras/clasificación , Venenos de Serpiente/enzimologíaRESUMEN
The genome of the sea anemone Nematostella vectensis (Nv) (Cnidaria, Anthozoa) was sequenced recently (Putnam et al., Science 317: 86, 2007). In the current study, 22 proteins of Nv were identified as putative phospholipases A(2) (PLA(2)) that showed up to 40-50% sequence identity with secreted or intracellular PLA(2)s including those of humans. Nv1-Nv6 PLA(2)s have identity with secreted human group (G)IB and GIIA PLA(2)s and PLA(2)s of the sea anemones Adamsia carciniopados and Urticina crassicornis. Nv7 and Nv8 PLA(2)s have identity with human and bee venom GIII PLA(2)s and Nv9 PLA(2) with GXIIA PLA(2). Nv10-Nv13 PLA(2)s show identity with GIX PLA(2) of Conus magus and bacterial PLA(2)s but no significant identity with any human PLA(2). Nv14 has identity with intracellular GIV PLA(2), Nv15 with GVII PLA(2), Nv16 and Nv17 with GVIII PLA(2), Nv18-Nv20 with GVI PLA(2), and Nv21 and Nv22 with patatin, respectively. The observations indicate that the cnidarian phospholipasome contains a rich array of orthologs of most types of animal PLA(2)s, and that many of the PLA(2)-driven vital functions prevail in these ancient metazoans. Cnidarian PLA(2)s may be considered as evolutionary precursors of PLA(2)s of higher animals.
RESUMEN
Arachidonic acid metabolites, the eicosanoids, are key mediators of allergen-induced airway inflammation and remodeling in asthma. The availability of free arachidonate in cells for subsequent eicosanoid biosynthesis is controlled by phospholipase A(2)s (PLA(2)s), most notably cytosolic PLA(2)-alpha. 10 secreted PLA(2)s (sPLA(2)s) have also been identified, but their function in eicosanoid generation is poorly understood. We investigated the role of group X sPLA(2) (sPLA(2)-X), the sPLA(2) with the highest in vitro cellular phospholipolysis activity, in acute and chronic mouse asthma models in vivo. The lungs of sPLA(2)-X(-/-) mice, compared with those of sPLA(2)-X(+/+) littermates, had significant reduction in ovalbumin-induced infiltration by CD4(+) and CD8(+) T cells and eosinophils, goblet cell metaplasia, smooth muscle cell layer thickening, subepithelial fibrosis, and levels of T helper type 2 cell cytokines and eicosanoids. These data direct attention to sPLA(2)-X as a novel therapeutic target for asthma.