Occurrence of Helicobacter spp. and Fecal Bacterial Contamination in High-altitude Aquatic Environments from the Andes.

The occurrence of Helicobacter spp. and fecal bacterial contamination was investigated in high-altitude environments from the Northeastern Andes of Venezuela. Helicobacter DNA was detected by PCR in streams, drinking and irrigation waters, and one culture from drinking water by the HP enrichment medium for selection of Helicobacter pylori, which displayed 98.98% homology to this pathogen based on 16S rRNA gene sequencing. FISH demonstrated predominant coccoid cells of the target bacteria indicative of the viable but nonculturable (VBNC) state in all water samples and HP cultures. Our work reveals for the first time Helicobacter spp. in waters from one of the highest places in the world. These results, together with the presence of fecal coliforms (2-160 MPN/100 mL) from the headwaters of rivers to drinking and irrigation waters, alert fecal contamination and epidemiological implications in this area of ecological and economic importance for the region.

Structural characterization and biological implications of sulfated N-glycans in a serine protease from the neotropical moth Hylesia metabus (Cramer [1775]) (Lepidoptera: Saturniidae).

Contact with the urticating setae from the abdomen of adult females of the neo-tropical moth Hylesia metabus gives rise to an urticating dermatitis, characterized by intense pruritus, generalized malaise and occasionally ocular lesions (lepidopterism). The setae contain a pro-inflammatory glycosylated protease homologous to other S1A serine proteases of insects. Deglycosylation with PNGase F in the presence of a buffer prepared with 40% H2 (18)O allowed the assignment of an N-glycosylation site. Five main paucimannosidic N-glycans were identified, three of which were exclusively α(1-6)-fucosylated at the proximal GlcNAc. A considerable portion of these N-glycans are anionic species sulfated on either the 4- or the 6-position of the α(1-6)-mannose residue of the core. The application of chemically and enzymatically modified variants of the toxin in an animal model in guinea pigs showed that the pro-inflammatory and immunological reactions, e.g. disseminated fibrin deposition and activation of neutrophils, are due to the presence of sulfate-linked groups and not on disulfide bonds, as demonstrated by the reduction and S-alkylation of the toxin. On the other hand, the hemorrhagic vascular lesions observed are attributed to the proteolytic activity of the toxin. Thus, N-glycan sulfation may constitute a defense mechanism against predators.

Evidence of Helicobacter spp. in freshwaters from Roraima Tepui, Guayana Shield, South America.

Helicobacter presence and viability in waters is not well characterized. The identification of natural reservoirs and infection sources may provide novel insights into its waterborne transmission. The goal of this study was to investigated the occurrence of Helicobacter spp. in natural freshwaters from Roraima Tepui, a little studied and unique ecosystem of the Guayana Shield. Freshwaters collected from two localities at Roraima Tepui were cultured in HP selective broth and agar for Helicobacter pylori and analysed by fluorescent in situ hybridization (FISH), specific PCR assays, 16S rRNA gene sequencing and phylogenetic analysis. The presence of other bacteria in freshwater enrichments was determined using clone library sequencing of the 16S rRNA gene and phylogenetic inferences. Helicobacter spp. were detected by semi-nested PCR and FISH in freshwater enrichments from both sites. Coccoid viable but nonculturable (VBNC) cells were evidenced using 16S rRNA gene Helicobacter species and H. pylori-specific probes. Partial 16S rRNA gene sequences of two HP enrichments showed high similarity to H. pylori and Helicobacter nemestrinae (99-100 %). Other bacteria such as Serratia, Aquitalea, Chromobacterium, Mycobacterium, Acinetobacter, Curvibacter and Dysgonomonas were also detected using complete 16S rRNA gene sequences, with Serratia, Aquitalea and Chromobacterium the most common genera (40.9, 18.2 and 15.2 %, respectively). This is the first time that Helicobacter spp. have been reported in freshwaters of a tepui ecosystem. Our results contribute to the current knowledge of these bacteria in the aquatic environment and expand their known/potential sites outside the human host.

[Clinical case of anaphylaxis due to eye drops]. / Caso clínico de anafilaxia por gotas oftálmicas.

BACKGROUND: Anaphylaxis is a severe systemic allergic reaction that can be life-threatening, timely diagnosis and treatment is required in these patients, one of the most frequent triggers is pharmacological. OBJECTIVE: To report the case of a patient who presented anaphylaxis due to eye drops. CASE REPORT: A 7-year-old male with a history of rhinitis and asthma with good control. It started with itchy eyes, ophthalmic drops were administered, composition: Polyethylene glycol 400, 0.4%, Propylene glycol 3 mg, polyquad 0.001%, presenting at 15 minutes an episode of anaphylaxis initially characterized by pruritus and intense conjunctival erythema, later nausea, vomiting, sweating, weakness, urticaria/facial angioedema and dyspnea were added, this episode was controlled opportunely with Levocetirizine 5 mg sublingual and Betametasona 4 mg intramuscular, progressively improving over the next 2 hours. The patient was evaluated by the Allergist, written recommendations were given to the mother in case this reaction occurred again, the use of the drops was prohibited, and the performance of skin test and a probable conjunctival provocation protocolized with the ophthalmic drops were pending. Accidentally 2 months later the patient was re-exposed with the same eye drops, presenting a similar reaction 15 minutes after the administration of the medication, they went to the emergency room where he received antihistamine and corticosteroid intravenous treatment, after this re-exposure is confirmed to the ophthalmic drops mentioned above as a trigger of anaphylaxis in this patient. CONCLUSIONS: We present a case of conjunctival anaphylaxis after application of eye drops, confirmed by re-exposure to the drug. It is essential to give diagnoses, recommendations with treatments and avoidance of the probable triggering agent of the reaction. The administration of immediate medication when the allergic episode begins in these patients can be vital, even more so when they live far from a health center, as was the case in this patient.

ANTECEDENTES: La anafilaxia es una reacción alérgica sistémica severa que puede llegar a comprometer la vida. Se requiere de un diagnóstico y tratamiento oportuno en estos pacientes, uno de los desencadenantes más frecuente es el farmacológico. OBJETIVO: Reportar el caso de un paciente que presentó anafilaxia a gotas oftálmicas. REPORTE DE CASO: Varón de siete años de edad con antecedentes de rinitis y asma con buen control. Inició con picor ocular, se le administraron gotas oftálmicas, composición: Polietilenglicol 400, 0,4%, Propilenglicol 3 mg, polyquad 0,001%, y a los 15 minutos presentó un episodio de anafilaxia caracterizado, inicialmente, por prurito y eritema conjuntival intenso; posteriormente, presentó náuseas, vómito, sudoración, debilidad, urticaria/angioedema facial y disnea. Este episodio fue controlado en el momento, con tratamiento de Levoceterizina 5 mg s.l. y Betametasona 4 mg i.m., con mejoría progresiva en las siguientes dos horas. El paciente fue evaluado por la especialidad de alergología. A su madre se dieron recomendaciones por escrito, por si se presentaba nuevamente la reacción. Se prohibió la utilización de las gotas, y quedó pendiente la realización de las pruebas cutáneas y una probable provocación conjuntival protocolizada con las gotas oftálmicas. Accidentalmente, dos meses después se reexpuso al paciente con las mismas gotas oftálmicas, y a los 15 minutos de la administración del medicamento, presentó una reacción similar, por lo que acudieron a emergencias donde recibió tratamiento antihistamínico y corticoides vía i.m.; tras esta reexposición, se confirma a las gotas oftálmicas mencionadas anteriomente, como desencadenantes de anafilaxia en el paciente. CONCLUSIONES: Presentamos un caso sobre anafilaxia por vía conjuntival tras aplicación de gotas oftálmicas, confirmado por la reexposición al fármaco. Es esencial dar diagnósticos, recomendaciones con tratamientos y evitar el probable agente desencadenante de la reacción. La administración de medicación inmediata cuando inicia el episodio alérgico en estos pacientes, puede ser vital, más aún cuando viven lejos de un centro de salud, como era el caso referenciado.

Schlieren measurements in the round cylinder of an optically accessible internal combustion engine.

This paper describes the design and experimental application of an optical system to perform schlieren measurements in the curved geometry of the cylinder of an optically accessible internal combustion engine. Key features of the system are a pair of cylindrical positive meniscus lenses, which keep the beam collimated while passing through the unmodified, thick-walled optical cylinder, and a pulsed, high-power light-emitting diode with narrow spectral width. In combination with a high-speed CMOS camera, the system is used to visualize the fuel jet after injection of hydrogen fuel directly into the cylinder from a high-pressure injector. Residual aberrations, which limit the system's sensitivity, are characterized experimentally and are compared to the predictions of ray-tracing software.

Enteric Nervous System: Identification of a Novel Neuronal Sensory Network in the Duodenal Epithelium.

The communication between the intestinal epithelium and the enteric nervous system has been considered indirect. Mechanical or chemical stimuli activate enteroendocrine cells inducing hormone secretion, which act on sub-epithelial nerve ends, activating the enteric nervous system. However, we identified an epithelial cell that expresses NKAIN4, a neuronal protein associated with the ß-subunit of Na+/K+-ATPase. This cell overexpresses Na+/K+-ATPase and ouabain-insensitive Na+-ATPase, enzymes involved in active sodium transport. NKAIN4-positive cells also express neuronal markers as NeuN, acetylcholine-esterase, acetylcholine-transferase, α3- and α7-subunits of ACh receptors, glutamic-decarboxylase, and serotonin-receptor-7, suggesting they are neurons. NKAIN4-positive cells show a polarized shape with an oval body, an apical process finished in a knob-like terminal in contact with the lumen, a basal cilia body at the base of the apical extension, and basal axon-like soma projections connecting sub-epithelial nerve terminals, lymphoid nodules, glial cells, and enterochromaffin cells, forming a network that reaches the epithelial surface. We also showed, using retrograde labeling and immunofluorescence, that these cells receive afferent signals from the enteric nervous system. Finally, we demonstrated that acetylcholine activates NKAIN4-positive cells inducing Ca2+ mobilization and probably serotonin secretion in enterochromaffin cells. NKAIN4-positive cells are neurons that would form a part of a duodenal sensory network for physiological or noxious luminal stimuli.

Alzheimer's Disease and Green Tea: Epigallocatechin-3-Gallate as a Modulator of Inflammation and Oxidative Stress.

Alzheimer's disease (AD) is the most common cause of dementia, characterised by a marked decline of both memory and cognition, along with pathophysiological hallmarks including amyloid beta peptide (Aß) accumulation, tau protein hyperphosphorylation, neuronal loss and inflammation in the brain. Additionally, oxidative stress caused by an imbalance between free radicals and antioxidants is considered one of the main risk factors for AD, since it can result in protein, lipid and nucleic acid damage and exacerbate Aß and tau pathology. To date, there is a lack of successful pharmacological approaches to cure or even ameliorate the terrible impact of this disease. Due to this, dietary compounds with antioxidative and anti-inflammatory properties acquire special relevance as potential therapeutic agents. In this context, green tea, and its main bioactive compound, epigallocatechin-3-gallate (EGCG), have been targeted as a plausible option for the modulation of AD. Specifically, EGCG acts as an antioxidant by regulating inflammatory processes involved in neurodegeneration such as ferroptosis and microglia-induced cytotoxicity and by inducing signalling pathways related to neuronal survival. Furthermore, it reduces tau hyperphosphorylation and aggregation and promotes the non-amyloidogenic route of APP processing, thus preventing the formation of Aß and its subsequent accumulation. Taken together, these results suggest that EGCG may be a suitable candidate in the search for potential therapeutic compounds for neurodegenerative disorders involving inflammation and oxidative stress, including Alzheimer's disease.

Bioaccumulation of Essential and Potentially Toxic Elements in the Muscle and Liver of the Spotted Ratfish (Hydrolagus colliei) From Deep-Sea Waters off the Northern Gulf of California.

This study aimed to establish the distribution of As, Cd, Cu, Pb, and Zn, in the muscle and liver of the spotted ratfish Hydrolagus colliei from the northern Gulf of California to establish the bioaccumulation background data in this species. The individuals (n = 110) were obtained by bycatch from the Gulf of California hake fisheries, and the metals and metalloid were measured by atomic absorption spectrometry. The element with the highest concentration in the muscle (15.19 ± 5.40 mg kg-1) and the liver (20.98 ± 10.30 mg kg-1) was As, followed by essential elements (Zn > Cu), and the lowest were the non-essential Pb (0.029 ± 0.014 and 0.048 ± 0.038 mg kg-1, muscle and liver, respectively) and Cd (0.022 ± 0.014 and 0.796 ± 0.495 mg kg-1, muscle and liver, respectively). The liver showed higher bioaccumulation than the muscle in all the studied elements. The sex was not a factor that influenced the bioaccumulation. The concentrations of As in the muscle did not exceed the maximum permissible limits of Mexican legislation, and < 50% of the samples exceed Cd and Pb limits of the Mexican, European Union, and WHO/FAO regulations. The differences found between the elements and tissues could be related to the different diets of the species, their migratory patterns, and their life conditions. Studies in the deep-sea water H. colliei are limited, and further investigations are needed regarding the feeding habits of H. colliei as well as the interactions of potentially toxic elements within the deep-sea water habitat.

Consequences of zinc deficiency on zinc localization, taurine transport, and zinc transporters in rat retina.

The colocalization of taurine and zinc transporters (TAUT, ZnTs) has not been explored in retina. Our objective is to evaluate the effect of the intracellular zinc chelator N,N,N,N-tetrakis-(2-pyridylmethyl) ethylenediamine (TPEN) on zinc localization and colocalization TAUT and ZnT-1 (of plasma membrane), 3 (vesicular), and 7 (vesicular and golgi apparatus) in layers of retina by immunohistochemistry. To mark zinc, it was used cell-permeable fluorescent Zinquin ethyl ester. Specific first and secondary antibodies, conjugated with rhodamine or fluorescein-isothiocyanate were used to mark TAUT and ZnTs. The fluorescence results were reported as integrated optical density (IOD). Zinc was detected in all layers of the retina. The treatment with TPEN produced changes in the distribution of zinc in layers of retina less in the outer nuclear layer compared with the control. TAUT was detected in all layers of retina and TPEN chelator produced decrease of IOD in all layers of retina except in the photoreceptor compared with the control. ZnT 1, 3, and 7 were distributed in all retina layers, with more intensity in ganglion cell layer (GCL) and in the layers where there is synaptic connection. For all transporters, the treatment with TPEN produced significant decrease of IOD in layers of retina least in the inner nuclear layer for ZnT1, in the photoreceptor for ZnT3 and in the GCL and outer plexiform layer for ZnT7. The distribution of zinc, TAUT, and ZnTs in the layers of retina is indicative of the interaction of taurine and zinc for the function of the retina and normal operation of said layers. HIGHLIGHTS: Taurine and zinc are two molecules highly concentrated in the retina and with relevant functions in this structure. Maintaining zinc homeostasis in this tissue is necessary for the normal function of the taurine system in the retina. The study of the taurine transporter and the different zinc transporters in the retina (responsible for maintaining adequate levels of taurine and zinc) is relevant and novel, since it is indicative of the interactions between both molecules in this structure.

Coral reefs at Sir Bu Nair Island: An offshore refuge of Acropora in the southern Arabian Gulf.

Coral reefs across the southern Arabian Gulf have declined in the past two decades, with extensive loss of formerly Acropora table corals, which are now functionally extinct in nearshore reefs. This study documents the coral community at Sir Bu Nair (SBN), an offshore island buffered by less extreme environmental conditions, which contains the last remaining large stands of Acropora in the southern Gulf. We found that Acropora is a major reef-building coral throughout SBN. Mean coral cover was 27% (range: 6%-49%) across all sites and depths, of which more than half was comprised by Acropora. This varied around the island, with the highest densities to the south and southwest in shallow waters. Our study provides essential information for the management and conservation of these highly valuable and vulnerable corals.

Expression of Paracoccidioides brasiliensis CHS3 in a Saccharomyces cerevisiae chs3 null mutant demonstrates its functionality as a chitin synthase gene.

We report the isolation and sequencing of CHS3, a gene that encodes one of several chitin synthases in Paracoccidioides brasiliensis, a medically important fungus restricted geographically to Latin America. The gene contains a single open reading frame of 3817 bp with two introns (71 and 86 bp) and encodes a 1220 amino acid polypeptide with high similarity to other fungal chitin synthases. Northern analysis reveals a high expression of CHS3 in the pathogenic yeast-like phase of the fungus and at the end of the mycelium-yeast transition. Expression of P. brasiliensis CHS3 in a Saccharomyces cerevisiae chs3 null mutant enhanced calcofluor white staining in parallel to an increase in total chitin synthase activity and chitin content in its cell wall.

Inhibition of cholinesterase activity by soil extracts and predicted environmental concentrations (PEC) to select relevant pesticides in polluted soils.

The correlation of predicted environmental concentrations (PEC) with cholinesterase activity inhibition detected in soil extracts was determined. PEC was derived from organophosphate (OP) and carbamate (CA) compounds applied to a flower crop area. Samples of surface soil (0 - 30 cm in depth) and subsurface soil (30 to 60 cm in depth) were taken from a flower crop area in which OP pesticides such as acephate ((RS)-N-[methoxy(methylthio)phosphinoyl]acetamide), dimethoate (2-dimethoxyphosphinothioylthio-N-methylacetamide) and methyl parathion (O,O-dimethyl O-4-nitrophenyl phosphorothioate), and CA pesticides such as carbendazim (methyl benzimidazol-2-ylcarbamate), carbofuran (2,3-dihydro-2,2-dimethylbenzofuran-7-yl methylcarbamate) and methomyl (S-methyl (EZ)-N-(methylcarbamoyloxy) thioacetimidate) were applied for two years. Weekly loads of these pesticides were registered to estimate the annual load of each compound. Physicochemical analysis and relative inhibition of cholinesterasic activity were measured for each soil sample. PEC values were estimated with Pesticide Analytical Model (PESTAN), a leach model, for each pesticide using soil sample data obtained from physicochemical analysis. From all pesticides tested, only acephate and methomyl showed a significant correlation (p < 0.01) between PEC values and inhibition cholinesterase activity of soil extracts. These results suggest that inhibition of cholinesterase activity observed in soil extracts is produced mainly by these two pesticides. Further studies could be developed to measure acephate and methomyl concentrations to reduce their environmental impact.

[Jejunal gastrointestinal stromal tumor (GIST): clinical case]. / Tumor del estroma gastrointestinal (GIST) de yeyuno: caso clínico.

Background: Gastrointestinal stromal tumors (GIST) are the most common mesenchymal tumors of the gastrointestinal tract. It is exposed a clinical case of jejunal GIST from a second-level hospital in Mexico. Clinical case: Female patient of 76 years, with history of tobacco use (two cigarettes per day for 25 years), that is referred to General Surgery due to a four month evolution of symptoms, characterized by abdominal pain, hyporexia and urinary symptomatology. Physical examination revealed a non-pulsatile, solid, non-mobile, non-painful mass in the hypogastrium and right iliac fossa of approximately 15 cm in length. Ovarian tumor was ruled out, since CEA and CA-125 tumor markers were negative. Abdominopelvic ultrasound was performed and reported a solid tumor with cystic spaces inside. CT reported a solid tumor of 9.5 x 2.5 x 8.3 cm, with defined edges, multilobed, presence of some calcifications in its wall that did not show enhancement with the use of contrast media. Patient underwent exploratory laparotomy and it was found a jejunal tumor, 210 cm from the ligament of Treitz. Immunohistochemistry reported positivity to KIT and DOG1, confirming the diagnosis of GIST. Conclusion: GISTs are uncommon entities. Their clinical presentation is insidious and the preoperative diagnosis is complex due to the need for biopsy. The treatment is surgery, but tyrosine kinase inhibitors should be administered. Even in patients with response to treatment, follow-up is mandatory due to the risk of recurrence.

Introducción: los tumores del estroma gastrointestinal (GIST) son los tumores mesenquimatosos más comunes del tracto gastrointestinal. Se expone un caso clínico de GIST en yeyuno que se presentó en un hospital de segundo nivel en México. Caso clínico: femenino de 76 años, con antecedente de tabaquismo (dos cigarros diarios durante 25 años), referida a Cirugía General por cuadro de cuatro meses de evolución (dolor abdominal tipo cólico en hipogastrio, hiporexia y sintomatología urinaria). A la exploración física, se le detectó tumor no pulsátil, sólido, no móvil, no doloroso, adherido a planos profundos en hipogastrio y fosa ilíaca derecha de aprox. 15 cm de longitud. Se descartó tumor ovárico al resultar negativos los marcadores tumorales ACE y CA-125. Se realizó ultrasonido abdominopélvico que reportó imagen de tumoración sólida con zonas quísticas en su interior. La TC reportó tumoración sólida, de bordes definidos, multilobulada con algunas calcificaciones milimétricas en su pared de 9.5 x 2.5 x 8.3 cm y sin realce al administrar medio de contraste. La paciente se sometió a laparotomía exploradora y se encontró tumoración adherida a yeyuno a 210 cm del ligamento de Treitz. El tumor fue positivo a KIT y DOG1, lo que confirmó el diagnóstico de GIST de patrón fusiforme. Conclusión: los GIST son poco frecuentes. Su presentación clínica es insidiosa y el diagnóstico preoperatorio es complejo debido a la toma de biopsia. El tratamiento continúa siendo la cirugía, pero se deben administrar inhibidores de la tirosina cinasa. Incluso en pacientes con respuesta favorable al tratamiento, se recomienda seguimiento por riesgo de recidiva.

Localization of taurine transporter, taurine, and zinc in goldfish retina.

Taurine and zinc interact during structural and functional development of the rat retina and during the process of regeneration of retinal fragments from goldfish. These observations formed the basis for evaluating the regional correlation between the localization of the taurine transporter, taurine content and labile zinc content in goldfish retina. In the retina, the taurine transporter is expressed in photoreceptors, the outer plexiform layer, the inner nuclear layer and the ganglion cell layer. Taurine was detected in photoreceptors, the outer and inner nuclear layers, the outer plexiform layer and the ganglion cell layer. A large amount of labile zinc was detected in photoreceptors and to a lesser extent in ganglion cells. The taurine transporter, taurine and zinc coexist in photoreceptors and the ganglion cell layer. Their co-localization in photoreceptors may be related to the neuro-protective role of taurine and zinc in this layer. By contrast, their co-existence in ganglion cells may be related to their involvement in cell differentiation, development, and regeneration. This reveals the importance of taurine and zinc in maintaining normal cellular function in these particular layers of the retina.

Autologous tumor cells/bacillus Calmette-Guérin/formalin-based novel breast cancer vaccine induces an immune antitumor response.

Autologous cancer cell vaccines represent a multivalent patient-specific treatment. Studies have demonstrated that these immunotherapies should be combined with immunomodulators to improve results. We tested in breast cancer the antitumor effects of a 200 µg autologous tumor cells homogenate combined with 0.0625 mg of bacillus Calmette-Guérin (BCG), and 0.02% formalin. We used a 4T1 murine model of BALB/c receiving four weekly injections of either this vaccine or control treatments. The control treatments were either Phosphate Buffer Saline, BCG treated with formalin, or the tumor cells homogenate plus BCG alone. We found that mice treated with the vaccine had the lowest tumor growth rate and mitosis percentage. The vaccinated group also showed a marked increase in infiltration of antitumor cells (natural killer, CD8 + T and CD4+ Th1 cells), as well as a decrease of myeloid-derived suppressor cells (MDSCs) and tumor-associated macrophages (TAMs). Additionally, we also observed a possible activation of the immune memory response as indicated by plasma cell tumor infiltration. Our results demonstrate that our proposed breast cancer vaccine induces a potent antitumor effect in 4T1 tumor-bearing mice. Its effectiveness, low cost and simple preparation method, makes it a promising treatment candidate for personalized breast cancer immunotherapy.

Initial volume of a drug before it reaches the volume of distribution: pharmacokinetics of F(ab')2 antivenoms and other drugs.

Fast disappearance of F(ab')2 antivenoms from the plasma compartment [Sevcik et al., 2004. Modelling Tityus scorpion venom and antivenom pharmacokinetics. Evidence of active immunoglobulin G's F(ab')2 extrusion mechanism from blood to tissues. Toxicon 44, 731-734; Vazquez et al., 2005. Pharmacokinetics of a F(ab')2 scorpion antivenom in healthy human volunteers. Toxicon 46, 797-805] suggests a quick time course to reach its final distribution volume. An equation was developed to describe how the volume occupied by a drug in the body grows with time. As discussed in the paper this equation is free of some shortcomings of an equation developed for the same purpose by Niazi [1976. Volume of distribution as a function of time. J. Pharm. Sci. 65, 452-454]. Fluorescence microscopy showed that the rapid initial decay in plasmatic F(ab')2 concentration may be related to uptake of F(ab')2 by vascular endothelium which, in combination with accumulation in the vascular wall connective tissue, may produce an intermediate plateau in F(ab')2 V(sl)(t), which reached its final value after 10 h. The V(sl)(t) equation predicts that the plasma concentration half-time of decay has little use to estimate how a drug distributes through the body to exert its action, and predicts that, in some instances, intermediate plateaus in the time course of V(sl)(t) exist. Data from the literature showed that the kinetic considerations for V(sl)(t) also apply to clevidipine, digoxin, digitoxin, lidocaine and thiopentone.

Effect of leukocyte inhibitors benzydamine and cyclophosphamide, on lung injury caused by Tityus discrepans scorpion venom.

We studied the effects of benzydamine (BZ) and cyclophosphamide (CP) on acute lung injury caused by Tityus discrepans venom. Mice (male, IVIC strain, approximately 25g) were pretreated with either BZ (20microg/g) or CP (100microg/g) i.p. or saline. Envenoming (2microg/g mouse) was induced sc. Lung fraction area occupied by fibrin (FF), nuclei (NF), alveolar space (AS) and parenchyma (PF) were determined. Venom increased FF, NF and PF, significantly, and decreased AS. BZ antagonised venom's effect on FF sharply, antagonised partially effects on PF and AS, but was not able to antagonise effect on NF. CP abolished venom effects on NF, AS and PF, but was not able to antagonise the effect on FF. CP was slightly less effective than BZ in reducing FF. Fibrin deposition was associated to leukocyte sequestration. Blocking pro-inflammatory leukocyte metabolic pathways with BZ diminished FF, suggesting that neutrophil activation, inflammation and coagulation are correlated in the genesis of scorpionism acute lung injury.

Isolation and partial characterization of proteins with vasodegenerative and proinflammatory properties from the egg-nests of Hylesia metabus (Lepidoptera: Saturniidae).

The Hylesia genus comprises a group of Neotropical moths ubiquitous in the Americas from Arizona to Argentina. One of the species of the Hylesia genus in Venezuela, French Guyana, and Trinidad has been identified as Hylesia metabus (Cramer 1775) (Lepidoptera: Saturniidae). In Venezuela, these moths are found in abundance in the mangrove (Avicennia spp.) swamps surrounding the Gulf of Paria and the Orinoco Delta in the eastern part of the country. During the mating season, the female adults shed copious amounts of urticating setae in the air, producing a severe papulovesicular dermatitis among the population in the affected areas. The females also use their urticating setae to protect the eggs during the hatching period. In the current study, we have isolated and partially characterized proteins with proinflammatory properties from the urticating setae in the egg-nests by using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and anionic exchange-high-performance liquid chromatography (HPLC). We also have studied the biological response of the egg-nest extract and the HPLC purified fractions by inoculation in guinea pigs; and, analyzing the tissue samples by means of histopathological methods. The results of this study show that the extracted venom and HPLC purified subcomponents give rise to an intense inflammatory reaction characterized by massive infiltration of inflammatory cells, echymoses, and vascular degeneration. Chromatographic separation showed that the venom was made up of proteins having selectively vasodegenerative-fibrinolytic or proinflammatory-quimotactic properties.

Thrombin selectively induces transcription of genes in human monocytes involved in inflammation and wound healing.

Thrombin is essential for blood coagulation but functions also as a mediator of cellular signalling. Gene expression microarray experiments in human monocytes revealed thrombin-induced upregulation of a limited subset of genes, which are almost exclusively involved in inflammation and wound healing. Among these, the expression of F3 gene encoding for tissue factor (TF) was enhanced indicating that this physiological initiator of coagulation cascade may create a feed-forward loop to enhance blood coagulation. Activation of protease-activated receptor type 1 (PAR1) was shown to play a main role in promoting TF expression. Moreover, thrombin induced phosphorylation of ERK1/2, an event that is required for expression of thrombin-regulated genes. Thrombin also increased the expression of TF at the protein level in monocytes as evidenced by Western blot and immunostaining. Furthermore, FXa generation induced by thrombin-stimulated monocytes was abolished by a TF blocking antibody and therefore it is entirely attributable to the expression of tissue factor. This cellular activity of thrombin provides a new molecular link between coagulation, inflammation and wound healing.