Indirect ELISA for Detection of Fascioliasis IgG Antibodies in Human Sera

Currently, coprological examination based on egg detection in stool samples is used as the most ideal standard for the diagnosis of human fascioliasis. However, this method has been proven not to be adequate when being employed in the acute phase of the disease, and presents a poor sensitivity during the chronic phase. Serodiagnosis has become an excellent alternative to coprological examination in efforts to combat the effects of fascioliasis on human and animal health. Human fascioliasis is usually recognized as an infection of the bile ducts and liver caused by Fasciola hepatica, known to affect over 2 million humans. In this research, Indirect Enzyme-linked Immunosorbent Assay (ELISA) was performed to discern between positive and negative IgG antibody titers in sera in collaboration with a lab in Peru. A batch of approximately 325 samples of human sera of endemic Fasciola hepatica from regions in Peru was gathered and sent to the laboratory in order to be examined with the method explained in the following paragraph. In the search for a test for the diagnosis of fascioliasis on humans, indirect ELISA started to be employed in order to determine positive and negative values. In the indirect ELISA test, the sample antibody is sandwiched between the antigen coated on the plate and an enzyme-labeled, anti-species globulin conjugate. The addition of an enzyme substrate-chromogen reagent causes color to develop. This color is directly proportional to the amount of bound sample antibody. The more antibody present in the sample, the stronger the color development in the test wells. Positive samples presented a very strong optical density value, while negative samples were clear or low optical density value, measured with the aid of a spectrophotometer

Development of an antibody-based capture enzyme-linked immunosorbent assay for detecting echinostoma caproni (trematoda) in experimentally infected rats: kinetics of coproantigen excretion

The present study reports on the development of a coproantigen capture enzyme-linked immunosorbent assay (ELISA) for detecting Echinostoma caproni in experimentally infected rats. The capture ELISA was based on polyclonal rabbit antibodies that recognize excretory–secretory (ES) antigens. The detection limit of pure ES was 3 ng/ml in sample buffer and 60 ng/ml in fecal samples. The test was evaluated using a follow-up of 10 rats experimentally infected with 100 metacercariae of E. caproni, and the results were compared with those of other diagnostic methods such as parasitological examination and antibody titers determined by indirect ELISA. Coproantigens were detected in all the infected rats from the first day postinfection (DPI). The period of maximal coproantigen excretion was between 7 and 21 DPI. The values remained positive until 49–56 DPI, coinciding with the disappearance of the eggs in the stool samples of the infected rats. The kinetics of coproantigen detection were correlated with those of egg output. The present assay provides an alternative tool for the diagnosis of the echinostome infections. The proposed capture ELISA makes possible an earlier diagnosis than that provided by parasitological examination and indirect ELISA and also allows for the differentiation of past and current infections. Our results show that this assay can also be used to monitor the course of echinostome infections.Toledo Navarro, Rafael, [email protected] ; Espert Fernandez, Ana M., [email protected] ; Marcilla Diaz, Antonio, [email protected] ; Esteban Sanchis, Jose Guillermo, [email protected]

The correlation between urinary 5-hydroxyindoleacetic acid and sperm quality in infertile men and rotating shift workers

<p>Abstract</p> <p>Background</p> <p>Serotonin is a neurotransmitter that modulates a wide range of neuroendocrine functions. However, excessive circulating serotonin levels may induce harmful effects in the male reproductive system. The objective of this study was to evaluate whether the levels of urinary 5-hydroxyindoleacetic acid (5-HIIA), a major serotonin metabolite, correlate with different classical seminal parameters.</p> <p>Methods</p> <p>Human ejaculates were obtained from 40 men attending infertility counselling and rotating shift workers by masturbation after 4-5 days of abstinence. Urinary 5- HIIA concentration was quantified by using a commercial ELISA kit. Forward motility was assessed by a computer-aided semen analysis (CASA) system. Sperm concentration was determined using the haemocytometer method. Sperm morphology was evaluated after Diff-Quik staining, while sperm vitality was estimated after Eosin-Nigrosin vital staining.</p> <p>Results</p> <p>Our results show that urinary 5-HIIA levels obtained from a set of 20 volunteers negatively correlated with sperm concentration, forward motility, morphology normal range and sperm vitality. On the other hand, we checked the relationship between male infertility and urinary 5-HIIA levels in 20 night shift workers. Thus, urinary 5-HIIA levels obtained from 10 recently-proven fathers were significantly lower than those found in 10 infertile males. Additionally, samples from recent fathers exhibited higher sperm concentration, as well as better forward motility and normal morphology rate.</p> <p>Conclusions</p> <p>In the light of our findings, we concluded that high serotonin levels, indirectly measured as urinary 5-HIIA levels, appear to play a role as an infertility determinant in male subjects.</p

Highly stable and efficient light-emitting electrochemical cells based on cationic iridium complexes bearing arylazole ancillary ligands

A series of bis-cyclometalated iridium(III) complexes of general formula [Ir(ppy)2(N∧N)][PF6] (ppy− = 2-phenylpyridinate; N∧N = 2-(1H-imidazol-2-yl)pyridine (1), 2-(2-pyridyl)benzimidazole (2), 1-methyl-2-pyridin-2-yl- 1H-benzimidazole (3), 2-(4′-thiazolyl)benzimidazole (4), 1- methyl-2-(4′-thiazolyl)benzimidazole (5)) is reported, and their use as electroluminescent materials in light-emitting electrochemical cell (LEC) devices is investigated. [2][PF6] and [3][PF6] are orange emitters with intense unstructured emission around 590 nm in acetonitrile solution. [1][PF6], [4][PF6], and [5][PF6] are green weak emitters with structured emission bands peaking around 500 nm. The different photophysical properties are due to the effect that the chemical structure of the ancillary ligand has on the nature of the emitting triplet state. Whereas the benzimidazole unit stabilizes the LUMO and gives rise to a 3MLCT/3LLCT emitting triplet in [2][PF6] and [3][PF6], the presence of the thiazolyl ring produces the opposite effect in [4][PF6] and [5][PF6] and the emitting state has a predominant 3LC character. Complexes with 3MLCT/3LLCT emitting triplets give rise to LEC devices with luminance values 1 order higher than those of complexes with 3LC emitting states. Protecting the imidazole N−H bond with a methyl group, as in complexes [3][PF6] and [5][PF6], shows that the emissive properties become more stable. [3][PF6] leads to outstanding LECs with simultaneously high luminance (904 cd m−2), efficiency (9.15 cd A−1), and stability (lifetime over 2500 h).Spanish Ministry of Economy and Competitiveness (MINECO) of Spain (projects CTQ2014- 58812-C2-1-R, MAT2014-55200, CTQ2014-55583-R, CTQ2014-61914-EXP, CTQ2015-71955-REDT, CTQ2015- 70371-REDT, CTQ2015-71154-P, and Unidad de Excelencia Marıá de Maeztu MDM-2015-0538), European Feder funds (CTQ2015-71154-P), Obra Social “la Caixa” (OSLC-2012- 007), Junta de Castilla y León (BU033-U16), and Generalitat Valenciana (Prometeo2016/135

Targets, Mechanisms and Cytotoxicity of Half-Sandwich Ir(III) Complexes Are Modulated by Structural Modifications on the Benzazole Ancillary Ligand

Cancers are driven by multiple genetic mutations but evolve to evade treatments targeting specific mutations. Nonetheless, cancers cannot evade a treatment that targets mitochondria, which are essential for tumor progression. Iridium complexes have shown anticancer properties, but they lack specificity for their intracellular targets, leading to undesirable side effects. Herein we present a systematic study on structure-activity relationships of eight arylbenzazole-based Iridium(III) complexes of type [IrCl(Cp*)], that have revealed the role of each atom of the ancillary ligand in the physical chemistry properties, cytotoxicity and mechanism of biological action. Neutral complexes, especially those bearing phenylbenzimidazole (HL1 and HL2), restrict the binding to DNA and albumin. One of them, complex 1[C,NH-Cl], is the most selective one, does not bind DNA, targets exclusively the mitochondria, disturbs the mitochondria membrane permeability inducing proton leak and increases ROS levels, triggering the molecular machinery of regulated cell death. In mice with orthotopic lung tumors, the administration of complex 1[C,NH-Cl] reduced the tumor burden. Cancers are more vulnerable than normal tissues to a treatment that harnesses mitochondrial dysfunction. Thus, complex 1[C,NH-Cl] characterization opens the way to the development of new compounds to exploit this vulnerabilityWe acknowledge the “la Caixa” Foundation (LCF/PR/PR12/11070003), Ministerio de Ciencia Innovación y Universidades-FEDER (RTI2018-102040-B-100) and Junta de Castilla y León-FEDER (BU305P18) for financial support. Networking support by COST Action CA18202 (NECTAR) is also acknowledged

First report of cucurbit chlorotic yellows virus infecting watermelon and zucchini in the Canary Islands, Spain

This work was funded by grants from Spanish Ministerio de Econolnia, industria y competitividad (RTA2017-00061-C03-02) and from Instituto Valenciano de Investigaciones Agrarias (IVlA) (51912), both co-funded by the European Regional Development Fund (ERDF).Alfaro Fernández, AO.; Espino De Paz, AI.; Botella-Guillen, M.; Font San Ambrosio, MI.; Sanauja, E.; Galipienso, L.; Rubio, L. (2022). First report of cucurbit chlorotic yellows virus infecting watermelon and zucchini in the Canary Islands, Spain. Plant Disease. 106(7):1-1. https://doi.org/10.1094/PDIS-10-21-2296-PDN11106

First report of Tomato torrado virus on weed hosts in Spain

Alfaro Fernández, AO.; Córdoba-Sellés, C.; Cebrián, M.; Herrera-Vásquez, J.; Sanchez Navarro, JA.; Juárez, M.; Espino, A.... (2088). First report of Tomato torrado virus on weed hosts in Spain. Plant Disease. 92(5):831-831. https://doi.org/10.1094/pdis-92-5-0831b83183192

PARÁMETROS BIOLÓGICOS DE APANTELES NR. ARISTOTELIAE (VIERECK) (HYMENOPTERA: BRACONIDAE), PARASITOIDE DEL ENROLLADOR DE LAS HOJAS, AMORBIA SP. (LEPIDOPTERA: TORTRICIDAE)

Apanteles nr. Aristoteliae (Viereck) (Hymenoptera: Braconidae) is an endoparasitoid of the leafroller Amorbia sp. (Lepdidoptera: Tortricidae) larvae. In this study, the life cycle, parasitism, sex ratio, and longevity of adults of this parasitoid were determined under laboratory conditions (22 ± 2 ºC, 60 ± 5% relative humidity, and a photoperiod of 16:8h [light:dark]). The life cycle of this parasitoid was 42.45 ± 0.76 days. The duration of development stages of egg-larva, pupa and adult was 14.04 ± 0.05, 9.22 ± 0.07, and 19.36 ± 0.74 days, respectively. Each A. nr. aristoteliae female parasitized 73.33 ± 7.55 Amorbia sp. larvae. The sex ratio of F1 generation was 95.1% males (n = 912). Longevity of adults was significantly different (U = 564, P = 0.01) between mated males (27 ± 2.55 days) and virgins (19.75 ± 0.70 days) but not (U = 182, P = 0.3) between mated females that were exposed to parasitization during all its life (26.4 ± 2.88 days), and virgins (23.13 ± 1.4 days). In order to obtain a better understanding about life parameters of A. nr. aristoteliae, future studies that include different rearing conditions (e.g., temperature and space) are needed.Apanteles nr. aristoteliae (Viereck) (Hymenoptera: Braconidae) es un endoparasitoide de larvas del enrollador de las hojas Amorbia sp. (Lepidoptera: Tortricidae). En este estudio se determinó, bajo condiciones de laboratorio (22 ± 2 ºC, 60 ± 5% de humedad relativa y un fotoperiodo de 16:8 h [luz: oscuridad]) el ciclo de vida, parasitismo, proporción sexual y longevidad de adultos de este parasitoide. La duración del ciclo de vida fue de 42.45 ± 0.76 días. El tiempo de desarrollo de los estados de huevolarva, pupa y adulto fue de 14.04 ± 0.05, 9.22 ± 0.07 y 19.36 ± 0.74 días, respectivamente. A lo largo de toda su vida, cada hembra de A. nr. aristoteliae parasitó 73.33 ± 7.55 larvas del enrollador de las hojas Amorbia sp. La proporción sexual de la generación F1 fue de 95.1% machos (n = 912). La longevidad de los adultos fue significativamente distinta (U = 564, P = 0.01) entre los machos apareados (27 ± 2.55 días) y vírgenes (19.75 ± 0.70 días), pero no (U = 182, P = 0.3) entre las hembras apareadas y que fueron sometidas a parasitación durante toda su vida (26.4 ± 2.88 días) y vírgenes (23.13 ± 1.4 días). Con el fin de obtener un mejor conocimiento de los parámetros de vida del parasitoide A. nr. aristoteliae, es necesario realizar futuros estudios que incluyan diferentes condiciones de cría (ej., temperatura y espacio)

Del color de los ojos al interior del genoma. Nuevas tecnologías aplicadas a la educación: una experiencia en la enseñanza de la Genética

Depto. de Genética, Fisiología y MicrobiologíaFac. de Ciencias BiológicasFALSEsubmitte

Reduced levels of intracellular calcium releasing in spermatozoa from asthenozoospermic patients

<p>Abstract</p> <p>Background</p> <p>Asthenozoospermia is one of the most common findings present in infertile males characterized by reduced or absent sperm motility, but its aetiology remains unknown in most cases. In addition, calcium is one of the most important ions regulating sperm motility. In this study we have investigated the progesterone-evoked intracellular calcium signal in ejaculated spermatozoa from men with normospermia or asthenozoospermia.</p> <p>Methods</p> <p>Human ejaculates were obtained from healthy volunteers and asthenospermic men by masturbation after 4–5 days of abstinence. For determination of cytosolic free calcium concentration, spermatozoa were loaded with the fluorescent ratiometric calcium indicator Fura-2.</p> <p>Results</p> <p>Treatment of spermatozoa from normospermic men with 20 micromolar progesterone plus 1 micromolar thapsigargin in a calcium free medium induced a typical transient increase in cytosolic free calcium concentration due to calcium release from internal stores. Similar results were obtained when spermatozoa were stimulated with progesterone alone. Subsequent addition of calcium to the external medium evoked a sustained elevation in cytosolic free calcium concentration indicative of capacitative calcium entry. However, when progesterone plus thapsigargin were administered to spermatozoa from patients with asthenozoospermia, calcium signal and subsequent calcium entry was much smaller compared to normospermic patients. As expected, pretreatment of normospermic spermatozoa with both the anti-progesterone receptor c262 antibody and with progesterone receptor antagonist RU-38486 decreased the calcium release induced by progesterone. Treatment of spermatozoa with cytochalasin D or jasplakinolide decreased the calcium entry evoked by depletion of internal calcium stores in normospermic patients, whereas these treatments proved to be ineffective at modifying the calcium entry in patients with asthenozoospermia.</p> <p>Conclusion</p> <p>Our results suggest that spermatozoa from asthenozoospermic patients present a reduced responsiveness to progesterone.</p