Zebrafish (Danio rerio) as a possible bioindicator of epigenetic factors present in drinking water that may affect reproductive function: is chorion an issue?

[EN] Emerging organic contaminants have been monitored in stream waters, raw and finished waters and wastewater effluents. Most of these contaminants, such as epigenetic substances, have been detected at very low levels. Unfortunately, their complete monitoring and/or removal are very difficult, given the increasing presence of new contaminants and due to analytical and economic considerations. For this reason, bioindicators are used as an alternative to monitor their presence. To this end, zebrafish is being used to assess certain contaminants in water quality studies. As our long-term aim is to determine if zebrafish (Danio rerio) can be used to detect environmental epigenetic factors in drinking waters with effects on human reproduction, an initial question is whether the chorion could interfere with the possible action of epigenetic factors in two reproductive events: genital ridge formation and migration of the primordial germ cells (PGCs) to these genital ridges. In the first experiment, we attempted to partially degrade the chorion of mid blastula transition (MBT) embryos with pronase, with acceptable survival rates at 5 days post fertilisation (dpf), with the group exposed for 15 min giving the best survival results. As denuded early embryos require a specific culture medium, in the next experiment embryo survival was evaluated when they were cultured up to 5 dpf in drinking waters from six different sources. Results showed a negative effect on embryo survival at 5 dpf from several waters but not in others, thus distorting the survival outcomes. These results suggest using embryos with the chorion intact from the outset when drinking waters from different sources are to be tested.Martínez-Sales, MI.; García Ximenez, F.; Espinos Gutierrez, FJ. (2015). Zebrafish (Danio rerio) as a possible bioindicator of epigenetic factors present in drinking water that may affect reproductive function: is chorion an issue?. Zygote. 23(3):447-452. doi:10.1017/S0967199414000045S44745223

Bronchopulmonary Penetration of Isavuconazole in Pulmonary Transplant Recipients (PBISA01): Protocol for a Phase IV Clinical Trial With a Single Treatment Arm

Background: Aspergillosis is the most frequently observed invasive fungal disease (IFD) in lung transplant recipients. Isavuconazole (ISA) has shown a better safety profile and noninferiority to voriconazole in the treatment of patients with IFD. Objective: The aim of this study is to describe the bronchopulmonary pharmacokinetic profile of oral ISA by analyzing the degree of penetration in the epithelial lining fluid and alveolar macrophages in patients receiving lung transplantation with a diagnosis of IFD. Methods: A total of 12 patients aged ≥18 years receiving a lung transplant with an IFD diagnosis and indication for ISA treatment and follow-up bronchoscopy will be included in the study. After 5 days of treatment with ISA and before the treatment is discontinued, the patients will be randomized (1:1:1:1) to perform the scheduled bronchoscopy at various times after the administration of ISA (2, 4, 8, and 12 hours). In total, 4 blood samples will be obtained per patient: at 72 hours after treatment initiation, on the day of the bronchoscopy, at the time of the bronchoalveolar lavage (simultaneously), and at 7 days after treatment initiation, to analyze tacrolimus and ISA plasma levels. ISA concentrations will be measured in plasma, epithelial lining fluid, and alveolar macrophages by a high-performance liquid chromatography/UV coupled to fluorescence method. Results: Enrollment for the PBISA01 trial began in October 2020 and was completed in October 2021. All samples will be analyzed once recruitment is complete, and the results are expected to be published in October 2022. Conclusions: There are no clinical studies that analyze the bronchopulmonary penetration of ISA. Bronchoalveolar lavage performed routinely in the follow-up of lung transplant recipients constitutes an opportunity to analyze the bronchopulmonary penetration of ISA. Trial registration: European Clinical Trials Register 2019-004240-30; www.clinicaltrialsregister.eu/ctr-search/trial/2019-004240-30/ES. International registered report identifier (irrid): DERR1-10.2196/37275.This work is supported funded by Pfizer (grant 54685521). Pfizer will have no role in the study’s design; the collection, management, analysis, and interpretation of data; writing of the report; and the decision to submit the report for publication.S

Bronchopulmonary penetration of isavuconazole in lung transplant recipients

Isavuconazole's (ISA) pharmacokinetics was studied among lung transplant recipients to evaluate its bronchopulmonary penetration. This study included 13 patients and showed mean serum concentrations of 3.30 (standard deviation [SD] 0.45), 5.12 (SD 1.36), and 6.31 (SD 0.95) at 2 h, 4 h, and 24 h respectively. Mean concentrations in the epithelial lining fluid were 0.969 (SD 0.895), 2.141 (SD 1.265), and 2.812 (SD 0.693) at the same time points. ISA is a drug with a tolerable safety profile that achieves adequate concentrations in the lung.This work was partially supported and funded by Pfizer (grant 54685521). Pfizer had no role in the study’s design; the collection, management, analysis, and interpretation of data; writing of the report; and the decision to submit the report for publicationS

Human Amebiasis: Breaking the Paradigm?

For over 30 years it has been established that the Entamoeba histolytica protozoan included two biologically and genetically different species, one with a pathogenic phenotype called E. histolytica and the other with a non-pathogenic phenotype called Entamoeba dispar. Both of these amoebae species can infect humans. E. histolytica has been considered as a potential pathogen that can cause serious damage to the large intestine (colitis, dysentery) and other extraintestinal organs, mainly the liver (amebic liver abscess), whereas E. dispar is a species that interacts with humans in a commensal relationship, causing no symptoms or any tissue damage. This paradigm, however, should be reconsidered or re-evaluated. In the present work, we report the detection and genotyping of E. dispar sequences of DNA obtained from patients with amebic liver abscesses, including the genotyping of an isolate obtained from a Brazilian patient with a clinical diagnosis of intestinal amebiasis that was previously characterized as an E. dispar species. The genetic diversity and phylogenetic analysis performed by our group has shown the existence of several different genotypes of E. dispar that can be associated to, or be potentiality responsible for intestinal or liver tissue damage, similar to that observed with E. histolytica

Comunicación corta. El desarrollo de embriones de pez cebra en estadío MBT puede ser detenido reversiblemente a 16 grados C

Germline chimaerism (intra or interspecific) is a technique of great potential in aquaculture. It allows specimens to be obtained that produce gametes whose origins lie in the cells of the donor organism. Chimaerism is usually performed at the mid blastula transition (MBT) stage since this is the last in which embryonic cells remain completely totipotent. Zebrafish are photoperiodic in their egg-laying behaviour and show rapid embryonic development. For chimaerism to be successful, it is of interest to establish the maximum time over which embryonic development can be reversibly arrested. This paper reports the effect on survival of subjecting zebrafish embryos at different stages of development to a water temperature of 16 deg C for different lengths of time. The maximum exposure time after which these embryos were able to resume development following low-temperature-induced developmental arrest became shorter as the embryonic stage exposed became earlier. At the MBT stage, the maximum safe exposure time was 2 h; longer exposure times led to problems in development and survival.El quimerismo intra e interespecífico en la línea germinal es una técnica con un gran potencial para la Acuicultura, ya que permite obtener especímenes en los que algunos gametos provienen de células (de la misma o de diferentes especies de interés) insertadas en el embrión. El quimerismo se efectúa habitualmente en el estadío MBT (mid blastula transition), ya que éste es el último estadío en el que las células embrionarias aún son totipotentes. El pez cebra es una especie de reproducción fotoperiódica y de rápido desarrollo embrionario, lo que hace de interés evaluar el tiempo y el estadío preciso en que es posible detener reversiblemente el desarrollo sin que éste resulte penalizado posteriormente. Se ha evaluado el efecto que ejerce la permanencia a 16 grados C sobre la supervivencia de embriones de pez cebra en diferentes estadíos. El tiempo máximo de exposición tras el cual los embriones pueden reanudar su desarrollo es tanto más limitado cuanto más temprano es su estadío. En el caso concreto de embriones de pez cebra en estadío MBT, éstos pueden ser mantenidos a 16 grados C durante 2 h como máximo sin penalizar su posterior supervivencia y desarrollo

PRELIMINARY RESULTS OF INTRA CYTOPLASMIC SPERM INJECTION (ICSI) APPLIED ON RABBIT : A TECHNICAL NOTE

[EN] In a foundation process of a maternal rabbit line using high selection intensities, from sorne hyper prolific females, selected as embryo donors, the subjects were very old and in an unhealthy condition (24%). An in vitro procedure to obtain embryos from this kind of females would be desirable. Sorne technical improvements used for human intracytoplasmic sperm injection (ICSI) have resulted in an enhancement of cleavage rates. This method may be applied to rabbits. Spermatozoa from ejaculated washed semen samples were immobilised in 5% PVP droplet by touching the tail and aspirated tail first into the microinjection pipette. The positioned oocyte was punctured. Sorne ooplasm was aspirated. After rupturing the plasmalema by gentle aspiration, the withdrawn ooplasm and the sperm were inserted into the oocyte. The cleavage rate of intact (86/107; 80%) oocytes was checked at 24 hours after ICSI (39/107; 36%). In vitro development up to the morulae (22/39; 56%) and blastocyst stage (5/39; 13%) were obtained.[FR] Chez le tapín, dans la procédure permettant d'obtenir une lignée matemeUe en uti/isant la sé/ection intensive á partir de femeUes tres prolifiques, sé/ectionnées comme donneuses d'embryons, les animaux deviennent trop llgés et sont dans de mauvaises conditions sanitaires (24%). 11 paraft préférable d'utiliser un procédé in vitro pour obtenir les embryons de teUes femeUes. Des améliorations techniques utilisées pour l'injection intracytop/asmique du sperme chez f'homme (ICSI) ont amé/ioré le taux de division. Cette méthode peut étre app/iquée chez le tapín. Les spermatozordes provenant d'échantiUons d'éjaculats lavés sont immobilisés, dans une goutte d'une so/ution contenant 5% de PVP, en touchant la queue et en les aspírant par la queue dans une micro pípette á injection. L'ovocyte repéré est ponctionné. Un peu d'ovop/asme est aspiré. Aprés avoir rompu le p/asmalema par une /égére aspiration, /'ovop/asme et le sperme sont injectés dans /'ovocyte. Le taux de division d'ovocytes intacts (861107 ; 80%) est contrO/e 24 heures apres l'/CSI (391107; 36%). Le développement in vitro jusqu'au stade morula (22139; 56%) et blastocyste (5139; 13%) a été atteint.This work was supported by CICYT AGF95-0850 and Conselleria de Educación y ciencia de la Comunidad Valenciana. We wish to thank Mr. Neil Macowan for help with the English version230Escribá, M.; García- Ximénez, F. (1999). PRELIMINARY RESULTS OF INTRA CYTOPLASMIC SPERM INJECTION (ICSI) APPLIED ON RABBIT : A TECHNICAL NOTE. World Rabbit Science. 06(2). https://doi.org/10.4995/wrs.1998.348SWORD22706

Effect of ovarian cystic or haemorrhagic follicles on embryo recovery and survival after transfer in hCG-ovulated rabbits

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