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CD300f immune receptor contributes to healthy aging by regulating inflammaging, metabolism, and cognitive decline

Cell Rep 2023 Oct 31;42(10):113269

Frances Evans 1 , Daniela Alí-Ruiz 2 , Natalia Rego 3 , María Luciana Negro-Demontel 1 , Natalia Lago 2 , Fabio Andrés Cawen 2 , Bruno Pannunzio 1 , Paula Sanchez-Molina 4 , Laura Reyes 5 , Andrea Paolino 5 , Jorge Rodríguez-Duarte 6 , Valentina Pérez-Torrado 7 , Almudena Chicote-González 8 , Celia Quijano 9 , Inés Marmisolle 9 , Ana Paula Mulet 10 , Geraldine Schlapp 10 , María Noel Meikle 10 , Mariana Bresque 7 , Martina Crispo 10 , Eduardo Savio 5 , Cristina Malagelada 8 , Carlos Escande 7 , Hugo Peluffo 11

1 Department of Histology and Embryology, Faculty of Medicine, UDELAR, Montevideo, Uruguay; Neuroinflammation and Gene Therapy Laboratory, Institut Pasteur de Montevideo, Montevideo, Uruguay. 2 Neuroinflammation and Gene Therapy Laboratory, Institut Pasteur de Montevideo, Montevideo, Uruguay. 3 Bioinformatics Unit, Institut Pasteur de Montevideo, Montevideo, Uruguay; Faculty of Sciences, UDELAR, Montevideo, Uruguay. 4 Department of Cell Biology, Physiology and Immunology, and Institute of Neuroscience, Universitat Autònoma de Barcelona, Barcelona, Spain. 5 Uruguayan Center for Molecular Imaging (CUDIM), Montevideo, Uruguay. 6 Laboratory of Vascular Biology and Drug Development, INDICYO Program, Institut Pasteur Montevideo, Montevideo, Uruguay. 7 Metabolic Diseases and Aging Laboratory, INDICYO Program, Institut Pasteur de Montevideo, Montevideo, Uruguay. 8 Unitat de Bioquímica i Biologia Molecular, Departament de Biomedicina, Facultat de Medicina i Ciències de la Salut, Universitat de Barcelona (UB), Barcelona, Spain; Institut de Neurociències, Universitat de Barcelona (UB), Barcelona, Spain. 9 Departamento de Bioquímica y Centro de Investigaciones Biomédicas (CEINBIO), Facultad de Medicina, Universidad de la República, Montevideo, Uruguay. 10 Unidad de Biotecnología en Animales de Laboratorio, Institut Pasteur de Montevideo, Montevideo, Uruguay. 11 Department of Histology and Embryology, Faculty of Medicine, UDELAR, Montevideo, Uruguay; Neuroinflammation and Gene Therapy Laboratory, Institut Pasteur de Montevideo, Montevideo, Uruguay; Unitat de Bioquímica i Biologia Molecular, Departament de Biomedicina, Facultat de Medicina i Ciències de la Salut, Universitat de Barcelona (UB), Barcelona, Spain; Institut de Neurociències, Universitat de Barcelona (UB), Barcelona, Spain. Electronic address: hugo.peluffo@ub.edu.

DOI: 10.1016/j.celrep.2023.113269
PMID: 37864797
Pubmed: https://pubmed.ncbi.nlm.nih.gov/37864797
Texto completo: https://linkinghub.elsevier.com/retrieve/pii/S2211-1247(23)01281-0

Abstract:
Emerging evidence suggests that immune receptors may participate in many aging-related processes such as energy metabolism, inflammation, and cognitive decline. CD300f, a TREM2-like lipid-sensing immune receptor, is an exceptional receptor as it integrates activating and inhibitory cell-signaling pathways that modulate inflammation, efferocytosis, and microglial metabolic fitness. We hypothesize that CD300f can regulate systemic aging-related processes and ultimately healthy lifespan. We closely followed several cohorts of two strains of CD300f-/- and WT mice of both sexes for 30 months and observed an important reduction in lifespan and healthspan in knockout mice. This was associated with systemic inflammaging, increased cognitive decline, reduced brain glucose uptake observed by 18FDG PET scans, enrichment in microglial aging/neurodegeneration phenotypes, proteostasis alterations, senescence, increased frailty, and sex-dependent systemic metabolic changes. Moreover, the absence of CD300f altered macrophage immunometabolic phenotype. Taken together, we provide strong evidence suggesting that myeloid cell CD300f immune receptor contributes to healthy aging.



Microglial CD300f immune receptor contributes to the maintenance of neuron viability in vitro and after a penetrating brain injury

Sci Rep 2023 Oct 5;13(1):16796

Daniela Alí-Ruiz 1 2 , Nathalia Vitureira 3 , Hugo Peluffo 4 5 6 7

1 Neuroinflammation and Gene Therapy Lab., Institut Pasteur de Montevideo, Montevideo, Uruguay. 2 Departamento de Histología y Embriología, Facultad de Medicina, UdelaR, Montevideo, Uruguay. 3 Departamento de Fisiología, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay. 4 Neuroinflammation and Gene Therapy Lab., Institut Pasteur de Montevideo, Montevideo, Uruguay. hugo.peluffo@pasteur.edu.uy. 5 Departamento de Histología y Embriología, Facultad de Medicina, UdelaR, Montevideo, Uruguay. hugo.peluffo@pasteur.edu.uy. 6 Unitat de Bioquímica i Biología Molecular, Departamento de Biomedicina, Facultat de Medicina i Ciències de la Salut, Universitat de Barcelona (UB), Barcelona, Spain. hugo.peluffo@pasteur.edu.uy. 7 Institut de Neurociències, Universitat de Barcelona (UB), Barcelona, Spain. hugo.peluffo@pasteur.edu.uy.

DOI: 10.1038/s41598-023-43840-1
PMID: 37798310
Pubmed: https://pubmed.ncbi.nlm.nih.gov/37798310
Texto completo: https://doi.org/10.1038/s41598-023-43840-1

Abstract:
Emerging evidences suggest that immune receptors participate in diverse microglial and macrophage functions by regulating their immunometabolism, inflammatory phenotype and phagocytosis. CD300f, a TREM2-like lipid sensing immune receptor, that integrates activating and inhibitory cell-signalling pathways, modulates inflammation, efferocytosis and microglial metabolic fitness. In particular, CD300f overexpression was described to be neuroprotective after an acute brain injury, suggesting a role for this immune receptor in neurotrophic interactions. Thus, we hypothesised that CD300f modulates neuronal survival through neuron-microglial interactions. In order to study its biological function, we used in vitro and in vivo approaches, CD300f-/- animals and rCD300f-Fc, a fusion protein that interrupts the endogen interaction between CD300f receptor-ligands. In hippocampal cocultures containing neurons and mixed glia, we observed that rCD300f-Fc, but not control IgGs induced neuronal death. In accordance, in vivo studies performed by injecting rCD300f-Fc or control IgGs into rat or WT or CD300 KO mice neocortex, showed an increased lesioned area after a penetrating brain injury. Interestingly, this neuronal death was dependent on glia, and the neurotoxic mechanism did not involve the increase of proinflammatory cytokines, the participation of NMDA receptors or ATP release. However, exogenous addition of glial cell line-derived neurotrophic factor (GDNF) prevented this process. Taken together, our results suggest that CD300f modulates neuronal survival in vitro and after a penetrating brain injury in vivo and that CD300f inhibition alters microglial phenotype generating a neurotoxic microenvironment.



Fertility preservation in male cancer patients. Counseling and reproductive outcomes

Front Cell Dev Biol 2023 Aug 16:11:1240152

Dana Kimelman 1 2 3 , Andrea Torrens 2 , Carla Bonelli 2 , Rossana Sapiro 4

1 Oncofertility Program, Centro Hospitalario Pereira Rossell, Administración de los Servicios de Salud del Estado (ASSE), Montevideo, Uruguay. 2 Reprovita Lab and Biobank, Montevideo, Uruguay. 3 Clínica Ginecotocológica "B", Facultad de Medicina, Universidad de la República, Montevideo, Uruguay. 4 Unidad Académica Histologia y Embriologia, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay.

DOI: 10.3389/fcell.2023.1240152
PMID: 37664467
Pubmed: https://pubmed.ncbi.nlm.nih.gov/37664467
Texto completo: https://doi.org/10.3389/fcell.2023.1240152

Abstract:
Introduction: Advances in cancer treatments have determined an increase in survival rates. However, these lifesaving therapies may have a negative impact on reproductive health. To diminish the infertility risk; different fertility preservation strategies have been designed. Sperm freezing is the gold standard fertility preservation method in the case of post-pubertal men. The main objective of this study is to evaluate the fertility status of Uruguayan male cancer survivors who have gone through sperm freezing, as well as to assess oncofertility counseling received by these patients. Methods: This is a descriptive, cross-sectional, observational, and transversal study. A survey was conducted on male cancer survivors who cryopreserved sperm between 1985 and 2021 in "Reprovita Lab and Biobank" which is the only sperm bank in this country. Results: One hundred thirty-five participants answered the survey. At the time of diagnosis, the mean age of patients was 28.8 ± 6.4 years old. Testicular was the most frequent type of cancer (64%). Only, 12% (n = 15) already had children at the time of diagnosis. Among the interviewed survivors, 50% (n = 62) attempted to conceive after cancer treatment, and 68% (n = 42) achieved natural pregnancy. Patients who did not achieve spontaneous conception (n = 11), used their cryopreserved samples, and 45.4% achieved pregnancy. About 86% (n = 107) of survivors believed that the timing of oncofertility referrals was appropriate and 97% considered that having the possibility of protecting their fertility was very important. Eighty percent (n = 101), were advised by their attending physicians, 14% (n = 18) sought advice from family members or friends, and 4% (n = 5) from oncofertility specialists. Discussion: To our knowledge, this is the first study evaluating the reproductive outcomes of male cancer survivors in our country and the region. Most of the interviewed survivors considered fertility preservation as a positive initiative, independent of their reproductive outcomes, reflecting the importance of fertility preservation counseling as one of the most important aspects for futurequality of life of young cancer patients.



Rod precursors in the adult retina of the Austrolebias charrua annual fish

Tissue Cell 2023 83:102150

M L Herrera 1 , S Silva 2 , I Berrosteguieta 2 , G Casanova 3 , J C Rosillo 4 , A S Fernández 5

1 Departamento Neurociencias Integrativas y Computacionales, Lab. Neurobiología Comparada, Instituto de Investigaciones Biológicas Clemente Estable (IIBCE), Avenida. Italia 3318, 11600 Montevideo, Uruguay; Sección Fisiología y Nutrición, Facultad de Ciencias, Universidad de la República, Iguá 4225, 11400 Montevideo, Uruguay. 2 Departamento Neurociencias Integrativas y Computacionales, Lab. Neurobiología Comparada, Instituto de Investigaciones Biológicas Clemente Estable (IIBCE), Avenida. Italia 3318, 11600 Montevideo, Uruguay. 3 Unidad de Microscopía Electrónica, Facultad de Ciencias, Universidad de la República, Iguá 4225, 11400 Montevideo, Uruguay. 4 Departamento Neurociencias Integrativas y Computacionales, Lab. Neurobiología Comparada, Instituto de Investigaciones Biológicas Clemente Estable (IIBCE), Avenida. Italia 3318, 11600 Montevideo, Uruguay; Departamento de Histología y Embriología, Facultad de Medicina, Universidad de la República, Avda. General Flores 2125, 11800 Montevideo, Uruguay. Electronic address: jrosillo@iibce.edu.uy. 5 Departamento Neurociencias Integrativas y Computacionales, Lab. Neurobiología Comparada, Instituto de Investigaciones Biológicas Clemente Estable (IIBCE), Avenida. Italia 3318, 11600 Montevideo, Uruguay; Laboratorio de Neurociencias, Facultad de Ciencias, Universidad de la República, Iguá 4225, 11400 Montevideo, Uruguay. Electronic address: afernandez@iibce.edu.uy.

DOI: 10.1016/j.tice.2023.102150
PMID: 37423033
Pubmed: https://pubmed.ncbi.nlm.nih.gov/37423033
Texto completo: https://www.sciencedirect.com/science/article/abs/pii/S0040816623001386?via%3Dihub

Abstract:
Rod photoreceptors in the adult teleost retina are produced by rod precursors located in the outer nuclear layer (ONL). Annual fishes of the genus Austrolebias exhibit extensive adult retinal cell proliferation and neurogenesis, as well as surprising adaptive strategies to their extreme and changing environment, including adult retinal plasticity. Thus, here we identify and characterize rod precursors in the ONL of the Austrolebias charrua retina. For this aim we used classical histological techniques, transmission electron microscopy, detection of cell proliferation, and immunohistochemistry. Through these complementary approaches, we describe a cell population clearly distinguishable from photoreceptors in the ONL of the adult retina of A. charrua, which we propose corresponds to the rod precursor population. These cells exhibited particular morphological and ultrastructural characteristics, uptake of cell proliferation markers (BrdU+) and expression of stem cell markers (Sox2+). Determining the existence of the population of rod precursors is crucial to understand the sequence of events related to retinal plasticity and regeneration.



Morphological evidence of the protective effects of a synthetic chalcone against the striatal myelin damage induced by glutaric acid

Int J Dev Neurosci 2023 83(3):274-296

Gabriela Casanova 1 2 3 , Juan Carlos Rosillo 2 4 , Marcie Jiménez 1 , Anabel Fernández 2 5 , Magela Rodao 1 , Gaby Martínez 1 , Eugenia Isasi 3 4 , Nadia Presa Gau 6 , Gabriel Otero 3 6 , Mauricio Cabrera 6 , Pablo Díaz-Amarilla 3 , Hugo Cerecetto 6 , Mercedes González 6 , Silvia Olivera-Bravo 3

1 Unidad de Microscopía Electrónica, Facultad de Ciencias, Universidad de la República (UdelaR), Montevideo, Uruguay. 2 Laboratorio de Neurobiología Comparada, Instituto de Investigaciones Biológicas Clemente Estable (IIBCE), Ministerio de Educación y Cultura (MEC), Montevideo, Uruguay. 3 Laboratorio de Neurobiología Celular y Molecular, IIBCE, MEC, Montevideo, Uruguay. 4 Departamento de Histología y Embriología, Facultad de Medicina, UdelaR, Montevideo, Uruguay. 5 Laboratorio de Neurociencias, Facultad de Ciencias, UdelaR, Montevideo, Uruguay. 6 Grupo de Química Orgánica Medicinal, Facultad de Ciencias, UdelaR, Montevideo, Uruguay.

DOI: 10.1002/jdn.10256
PMID: 37073624
Pubmed: https://pubmed.ncbi.nlm.nih.gov/37073624
Texto completo: https://doi.org/10.1002/jdn.10256

Abstract:
Ultrastructural features of striatal white matter and cells in an in vivo model of glutaric acidemia type I created by intracerebral injection of glutaric acid (GA) were analyzed by transmission electron microscopy and immunohistochemistry. To test if the white matter damage observed in this model could be prevented, we administered the synthetic chemopreventive molecule CH38 ((E)-3-(4-methylthiophenyl)-1-phenyl-2-propen-1-one) to newborn rats, previous to an intracerebroventricular injection of GA. The study was done when striatal myelination was incipient and when it was already established (at 12 and 45 days post-injection [DPI], respectively). Results obtained indicate that that the ultrastructure of astrocytes and neurons did not appear significantly affected by the GA bolus. Instead, in oligodendrocytes, the most prominent GA-dependent injury defects included endoplasmic reticulum (ER) stress and nuclear envelope swelling at 12 DPI. Altered and reduced immunoreactivities against heavy neurofilament (NF), proteolipid protein (PLP), and myelin-associated glycoprotein (MAG) together with axonal bundle fragmentation and decreased myelin were also found at both ages analyzed. CH38 by itself did not affect striatal cells or axonal packages. However, the group of rats that received CH38 before GA did not show evidence neither of ER stress nor nuclear envelope dilation in oligodendrocytes, and axonal bundles appeared less fragmented. In this group, labeling of NF and PLP was similar to the controls. These results suggest that the CH38 molecule is a candidate drug to prevent or decrease the neural damage elicited by a pathological increase of GA in the brain. Optimization of the treatments and identification of the mechanisms underlying CH38 protective effects will open new therapeutic windows to protect myelin, which is a vulnerable target of numerous nervous system diseases.



Computational and mitochondrial functional studies of novel compound heterozygous variants in SPATA5 gene support a causal link with epileptogenic encephalopathy

Hum Genomics 2023 17(1):14

Víctor Raggio 1 , Martín Graña 2 , Erik Winiarski 3 , Santiago Mansilla 4 5 , Camila Simoes 2 6 , Soledad Rodríguez 1 , Mariana Brandes 2 , Alejandra Tapié 1 , Laura Rodríguez 1 , Lucía Cibils 7 , Martina Alonso 5 8 , Jennyfer Martínez 5 8 , Tamara Fernández-Calero 2 9 , Fernanda Domínguez 1 10 , Melania Rosas Mezquida 7 , Laura Castro 5 8 , Alfredo Cerisola 7 , Hugo Naya 2 11 , Adriana Cassina 5 8 , Celia Quijano 5 8 , Lucía Spangenberg 12 13

1 Departamento de Genética, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay. 2 Bioinformatics Unit, Institut Pasteur de Montevideo, Montevideo, Uruguay. 3 Departamento de Histología y Embriología, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay. 4 Departamento de Métodos Cuantitativos, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay. 5 Centro de Investigaciones Biomédicas (CEINBIO), Universidad de la República, Montevideo, Uruguay. 6 Departamento Básico de Medicina, Hospital de Clínicas, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay. 7 Departamento de Neuropediatría, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay. 8 Departamento de Bioquímica, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay. 9 Department of Exact and Natural Sciences, Universidad Católica del Uruguay, 11600, Montevideo, Uruguay. 10 Universidad Católica del Uruguay, 11600, Montevideo, Uruguay. 11 Departamento de Producción Animal y Pasturas, Facultad de Agronomía, Universidad de la República, Montevideo, Uruguay. 12 Bioinformatics Unit, Institut Pasteur de Montevideo, Montevideo, Uruguay. lucia@pasteur.edu.uy. 13 Departamento Básico de Medicina, Hospital de Clínicas, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay. lucia@pasteur.edu.uy.

DOI: 10.1186/s40246-023-00463-x
PMID: 36849973
Pubmed: https://pubmed.ncbi.nlm.nih.gov/36849973
Texto completo: https://humgenomics.biomedcentral.com/articles/10.1186/s40246-023-00463-x

Abstract:
The SPATA5 gene encodes a 892 amino-acids long protein that has a putative mitochondrial targeting sequence and has been proposed to function in maintenance of mitochondrial function and integrity during mouse spermatogenesis. Several studies have associated homozygous or compound heterozygous mutations in SPATA5 gene to microcephaly, intellectual disability, seizures and hearing loss. This suggests a role of the SPATA5 gene also in neuronal development. Recently, our group presented results validating the use of blood cells for the assessment of mitochondrial function for diagnosis and follow-up of mitochondrial disease, minimizing the need for invasive procedures such as muscle biopsy. In this study, we were able to diagnose a patient with epileptogenic encephalopathy using next generation sequencing. We found two novel compound heterozygous variants in SPATA5 that are most likely causative. To analyze the impact of SPATA5 mutations on mitochondrial functional studies directly on the patients' mononuclear cells and platelets were undertaken. Oxygen consumption rates in platelets and PBMCs were impaired in the patient when compared to a healthy control. Also, a decrease in mitochondrial mass was observed in the patient monocytes with respect to the control. This suggests a true pathogenic effect of the mutations in mitochondrial function, especially in energy production and possibly biogenesis, leading to the observed phenotype.



SOD1G93A Astrocyte-Derived Extracellular Vesicles Induce Motor Neuron Death by a miRNA-155-5p-Mediated Mechanism

ASN Neuro 2023 Jan-Dec:15:17590914231197527

Soledad Marton 1 , Ernesto Miquel 1 , Joaquín Acosta-Rodríguez 1 , Santiago Fontenla 2 , Gabriela Libisch 3 , Patricia Cassina 1

1 Departamento de Histología y Embriología, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay. 2 Departamento de Genética, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay. 3 Laboratorio Hospedero Patógeno/UBM, Institut Pasteur, Montevideo, Uruguay.

DOI: 10.1177/17590914231197527
PMID: 37644868
Pubmed: https://pubmed.ncbi.nlm.nih.gov/37644868
Texto completo: https://www.tandfonline.com/doi/10.1177/17590914231197527

Abstract:
Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by upper and lower motor neuron (MN) degeneration. Astrocytes surrounding MNs are known to modulate ALS progression. When cocultured with astrocytes overexpressing the ALS-linked mutant Cu/Zn superoxide dismutase (SOD1G93A) or when cultured with conditioned medium from SOD1G93A astrocytes, MN survival is reduced. The exact mechanism of this neurotoxic effect is unknown. Astrocytes secrete extracellular vesicles (EVs) that transport protein, mRNA, and microRNA species from one cell to another. The size and protein markers characteristic of exosomes were observed in the EVs obtained from cultured astrocytes, indicating their abundance in exosomes. Here, we analyzed the microRNA content of the exosomes derived from SOD1G93A astrocytes and evaluated their role in MN survival. Purified MNs exposed to SOD1G93A astrocyte-derived exosomes showed reduced survival and neurite length compared to those exposed to exosomes derived from non-transgenic (non-Tg) astrocytes. Analysis of the miRNA content of the exosomes revealed that miR-155-5p and miR-582-3p are differentially expressed in SOD1G93A exosomes compared with exosomes from non-Tg astrocytes. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis indicates that miR-155-5p and miR-582-3p predicted targets are enriched in the neurotrophin signaling pathway. Importantly, when levels of miR-155-5p were reduced by incubation with a specific antagomir, SOD1G93A exosomes did not affect MN survival or neurite length. These results demonstrate that SOD1G93A-derived exosomes are sufficient to induce MN death, and miRNA-155-5p contributes to this effect. miRNA-155-5p may offer a new therapeutic target to modulate disease progression in ALS.

Icono PDF 2023_Marton_SOD1G93A Astrocyte-Derived Extracellular Vesicles Induce Motor Neuron Death by a miRNA-155-5p-Mediated Mechanism.pdf



Structural and functional changes in rat uterus induced by neonatal androgenization

J Mol Histol 2022 53(6):903-914

Rebeca Chávez-Genaro 1 , Agustina Toledo 1 , Karina Hernández 1 , Gabriel Anesetti 2

1 Laboratorio de Biología de la Reproducción, Departamento de Histología y Embriología, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay. 2 Laboratorio de Biología de la Reproducción, Departamento de Histología y Embriología, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay. ganesett@fmed.edu.uy.

DOI: 10.1007/s10735-022-10106-5
PMID: 36201133
Pubmed: https://pubmed.ncbi.nlm.nih.gov/36201133
Texto completo: https://doi.org/10.1007/s10735-022-10106-5

Abstract:
Fetal or neonatal androgen exposure has a programming effect on ovarian function inducing a polycystic ovarian syndrome-like condition. Its effects on uterine structure and function are poorly studied. The aim of this work was to characterize the temporal course of changes in the rat uterine structure induced by neonatal exposure to aromatizable or not aromatizable androgens. Rats were daily treated with testosterone, dihydrotestosterone or vehicle during follicle assembly period (postnatal days 1 to 5). Uterine histoarchitecture, hormonal milieu, endometrial stromal collagen and capillary density were analyzed at prepubertal, pubertal and adult ages. Our data shows that neonatal androgen exposure induces early and long-lasting deleterious effects on uterine development, including altered adenogenesis and superficial epithelial alterations and suggest a role for altered serum estradiol levels in the maintenance and worsening of the situation. Our results suggest that alterations of the neonatal androgenic environment on the uterus could be responsible for alterations in the processes of implantation and maintenance of the embryo in women with polycystic ovary syndrome.



Phasor-based multi-harmonic unmixing for in-vivo hyperspectral imaging

Methods Appl Fluoresc 2022 11(1)

Alexander Vallmitjana 1 , Paola Lepanto 2 , Florencia Irigoin 2 3 4 , Leonel Malacrida 4 5

1 Department of Biomedical Engineering, University of California, Irvine, Irvine, CA, United States of America. 2 Human Molecular Genetics Lab, Institut Pasteur de Montevideo, Montevideo, Mataojo 2020, CP11400, Uruguay. 3 Departamento de Histología y Embriología, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay. 4 Advanced Bioimaging Unit, Institut Pasteur of Montevideo and Universidad de la República, Montevideo, Uruguay. 5 Departamento de Fisiopatología, Hospital de Clínicas, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay.

DOI: 10.1088/2050-6120/ac9ae9
PMID: 36252561
Pubmed: https://pubmed.ncbi.nlm.nih.gov/36252561
Texto completo: https://doi.org/10.1088/2050-6120/ac9ae9

Abstract:
Hyperspectral imaging (HSI) is a paramount technique in biomedical science, however, unmixing and quantification of each spectral component is a challenging task. Traditional unmixing relies on algorithms that need spectroscopic parameters from the fluorescent species in the sample. The phasor-based multi-harmonic unmixing method requires only the empirical measurement of the pure species to compute the pixel-wise photon fraction of every spectral component. Using simulations, we demonstrate the feasibility of the approach for up to 5 components and explore the use of adding a 6th unknown component representing autofluorescence. The simulations show that the method can be successfully used in typical confocal imaging experiments (with pixel photon counts between 101and 103). As a proof of concept, we tested the method in living cells, using 5 common commercial dyes for organelle labeling and we easily and accurately separate them. Finally, we challenged the method by introducing a solvatochromic probe, 6-Dodecanoyl-N,N-dimethyl-2-naphthylamine (LAURDAN), intended to measure membrane dynamics on specific subcellular membrane-bound organelles by taking advantage of the linear combination between the organelle probes and LAURDAN. We succeeded in monitoring the membrane order in the Golgi apparatus, Mitochondria, and plasma membrane in the samein-vivocell and quantitatively comparing them. The phasor-based multi-harmonic unmixing method can help expand the outreach of HSI and democratize its use by the community for it does not require specialized knowledge.



Standards in semen examination: publishing reproducible and reliable data based on high-quality methodology

Hum Reprod 2022 37(11):2497-2502

Lars Björndahl 1 , Christopher L R Barratt 2 , David Mortimer 3 , Ashok Agarwal 4 , Robert J Aitken 5 , Juan G Alvarez 6 7 , Natalie Aneck-Hahn 8 , Stefan Arver 1 , Elisabetta Baldi 9 , Lluís Bassas 10 , Florence Boitrelle 11 12 , Riana Bornman 13 , Douglas T Carrell 14 15 , José A Castilla 16 17 , Gerardo Cerezo Parra 18 , Jerome H Check 19 20 , Patricia S Cuasnicu 21 , Sally Perreault Darney 22 23 , Christiaan de Jager 24 , Christopher J De Jonge 25 , Joël R Drevet 26 , Erma Z Drobnis 27 , Stefan S Du Plessis 28 , Michael L Eisenberg 29 30 , Sandro C Esteves 31 32 33 , Evangelini A Evgeni 34 35 , Alberto Ferlin 36 , Nicolas Garrido 37 , Aleksander Giwercman 38 , Ilse G F Goovaerts 39 , Trine B Haugen 40 , Ralf Henkel 41 42 , Lars Henningsohn 43 44 , Marie-Claude Hofmann 45 , James M Hotaling 46 , Piotr Jedrzejczak 47 , Pierre Jouannet 48 , Niels Jørgensen 49 50 , Jackson C Kirkman Brown 51 52 53 , Csilla Krausz 54 , Maciej Kurpisz 55 , Ulrik Kvist 1 , Dolores J Lamb 56 , Hagai Levine 57 , Kate L Loveland 58 , Robert I McLachlan 59 , Ali Mahran 60 61 , Liana Maree 42 , Sarah Martins da Silva 2 , Michael T Mbizvo 62 , Andreas Meinhardt 63 , Roelof Menkveld 64 , Sharon T Mortimer 3 65 , Sergey Moskovtsev 66 67 , Charles H Muller 68 , Maria José Munuce 69 , Monica Muratori 54 , Craig Niederberger 70 71 , Cristian O'Flaherty 72 , Rafael Oliva 73 74 , Willem Ombelet 75 76 , Allan A Pacey 77 78 , Michael A Palladino 79 , Ranjith Ramasamy 80 , Liliana Ramos 81 , Nathalie Rives 82 , Eduardo Rs Roldan 83 , Susan Rothmann 84 , Denny Sakkas 85 , Andrea Salonia 86 87 , Maria Cristina Sánchez-Pozo 88 , Rosanna Sapiro 89 , Stefan Schlatt 90 , Peter N Schlegel 91 , Hans-Christian Schuppe 92 , Rupin Shah 93 , Niels E Skakkebæk 49 , Katja Teerds 94 , Igor Toskin 95 , Herman Tournaye 96 , Paul J Turek 97 , Gerhard van der Horst 98 99 100 , Monica Vazquez-Levin 101 , Christina Wang 102 103 , Alex Wetzels 104 , Theodosia Zeginiadou 105 106 , Armand Zini 107

1 ANOVA, Karolinska University Hospital and Karolinska Institutet, Stockholm, Sweden. 2 Reproductive Medicine Research Group, Division of Systems Medicine, School of Medicine, Ninewells Hospital and Medical School, University of Dundee, Dundee, UK. 3 Oozoa Biomedical Inc., West Vancouver, BC, Canada. 4 Case Western Reserve University, Moreland Hills, OH, USA. 5 Priority Research Centre for Reproductive Science, Faculty of Science and Faculty of Health & Medicine, University of Newcastle, Callaghan, NSW, Australia. 6 Centro Androgen, La Coruña, Spain. 7 Harvard Medical School, Boston, MA, USA. 8 Department of Urology, University of Pretoria, Pretoria, South Africa. 9 Department of Experimental and Clinical Medicine, University of Florence, Florence, Tuscany, Italia. 10 Andrology Department, Laboratory of Andrology and Sperm Bank, Fundació Puigvert, Barcelona, Spain. 11 Department of Reproductive Biology, Fertility Preservation, Andrology, CECOS, Poissy Hospital, Poissy, France. 12 Paris Saclay University, UVSQ, INRAE, BREED, Jouy-en-Josas, France. 13 School of Health Systems and Public Health, University of Pretoria, Pretoria, South Africa. 14 Andrology and IVF Laboratory, Division of Urology, Department of Surgery, University of Utah School of Medicine, Salt Lake City, UT, USA. 15 Department of Human Genetics, University of Utah School of Medicine, Salt Lake City, UT, USA. 16 GAMETIA Biobank, Granada, Spain. 17 Hospital Universitario Virgen de las Nieves and Instituto de Investigación Biosanitaria ibs. GRANADA, Granada, Spain. 18 LAFER Sperm Bank, Tuxpan 10-606, Roma Sur, C.P. 06760, Cuauhtémoc, Mexico City, Mexico. 19 Robert Wood Johnson Medical School at Camden, The University of Medicine and Dentistry of New Jersey, Camden, NJ, USA. 20 Department of Obstetrics and Gynecology, Division of Reproductive Endocrinology & Infertility, Cooper Hospital/University Medical Center, Melrose Park, PA, USA. 21 Instituto de Biología y Medicina Experimental (IbyME-CONICET), Ciudad Autónoma de Buenos Aires, Buenos Aires, Argentina. 22 US EPA, Cary, NC, USA. 23 US NIH, Cary, NC, USA. 24 Faculty of Health Science, University of Pretoria, Pretoria, South Africa. 25 University of Minnesota Medical Center, University of Minnesota, Minneapolis, MN, USA. 26 Université Clermont Auvergne/CNRS/INSERM-GreD Institute, Clermont-Ferrand, France. 27 School of Medicine, University of Missouri, Columbia, MI, USA. 28 College of Medicine, Mohammed Bin Rashid University of Medicine and Health Sciences, Dubai, United Arab Emirates. 29 Male Reproductive Medicine and Surgery, Stanford University School of Medicine, Stanford, CA, USA. 30 Department of Urology, Stanford University School of Medicine, Stanford, CA, USA. 31 ANDROFERT, Andrology and Human Reproduction Clinic, Campinas, Brazil. 32 Department of Surgery (Division of Urology), University of Campinas (UNICAMP), Campinas, Brazil. 33 Faculty of Health, Aarhus University, Aarhus C, Denmark. 34 CRYOGONIA Cryopreservation Bank, Athens, Greece. 35 Laboratory of Physiology, Department of Medicine, Democritus University of Thrace, Greece. 36 Unit of Andrology and Reproductive Medicine, Department of Medicine, University of Padova, Padova, Italia. 37 IVI Foundation, Health Research Institute La Fe, Valencia, Spain. 38 Department of Translational Medicine, Lund University, Malmö, Sweden. 39 Antwerp University Hospital, Edegem, Belgium. 40 Department of Life Sciences and Health, Oslo Metropolitan University, Oslo, Norway. 41 Department of Metabolism, Digestion & Reproduction, Imperial College London, London, UK. 42 Department of Medical Bioscience, University of the Western Cape, Bellville, South Africa. 43 Division of Urology, Department of CLINTEC, Karolinska Institutet, Stockholm, Sweden. 44 Department of Urology, Karolinska University Hospital, Stockholm, Sweden. 45 Department of Endocrine Neoplasia & Hormonal Disorders, University of Texas MD Anderson Cancer Center, Houston, TX, USA. 46 Division of Urology, Department of Surgery, University of Utah School of Medicine, Salt Lake City, UT, USA. 47 Department of Cell Biology, Poznan University of Medical Science, Poznan, Poland. 48 Université Paris Descartes, Paris, France. 49 Department of Growth and Reproduction, Copenhagen University Hospital, Rigshospitalet, Copenhagen, Denmark. 50 International Center for Research and Research Training in Endocrine Disruption of Male Reproduction and Child Health (EDMaRC), Copenhagen University Hospital, Rigshospitalet, Copenhagen, Denmark. 51 Centre for Human Reproductive Science (ChRS), UK. 52 College of Medical & Dental Sciences, University of Birmingham, UK. 53 Birmingham Women's and Children's NHS Foundation Trust, UK. 54 Department of Biomedical, Experimental and Clinical Sciences "Mario Serio", University of Florence, Florence, Italy. 55 Department of Reproductive Biology and Stem Cells, Institutet of Human Genetics, Poznan, Poland. 56 Brady Department of Urology, Center for Reproductive Genomics and Englander Institute for Precision Medicine, Weill Cornell Medical College, Cornell University, New York, NY, USA. 57 Braun School of Public Health and Community Medicine, Hadassah Medical Center, The Faculty of Medicine, Hebrew University of Jerusalem, Jerusalem, Israel. 58 Hudson Institute, Centre for Reproductive Health, Monash University, Clayton, VIC, Australia. 59 Hudson Institute of Medical Research, Centre for Endocrinology and Metabolism, Monash University, Clayton, VIC, Australia. 60 Dermatology and Andrology Department, Assiut University Hospital, Assiut, Egypt. 61 Faculty of Medicine, Assiut University, Assiut, Egypt. 62 Country Director, Population Council (International Programs). 63 Department of Anatomy and Cell Biology, Justus-Liebig-University of Giessen, Giessen, Germany. 64 Department of Obstetrics and Gynaecology, Faculty of Medicine and Health Sciences, Stellenbosch University, Tygerberg, South Africa. 65 Division of REI, Department of Obstetrics & Gynaecology, Faculty of Medicine, University of British Columbia, Vancouver, BC, Canada. 66 Department of Laboratory Medicine & Pathobiology, University of Toronto, Toronto, ON, Canada. 67 CreATe Fertility Centre, Toronto, ON, Canada. 68 Male Fertility Laboratory, Department of Urology, University of Washington School of Medicine, Seattle, WA, USA. 69 Laboratorio de Medicina Reproductiva, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, Argentina. 70 Department of Urology, UIC College of Medicine, IL, USA. 71 Department of Bioengineering, UIC College of Engineering, IL, USA. 72 Department of Surgery (Urology Division), McGill University, Montréal, QC, Canada. 73 Molecular Biology of Reproduction and Development Group, Biomedical Research Institute August Pi I Sunyer (IDIBAPS), University of Barcelona, Barcelona, Spain. 74 Hospital Clínic, University of Barcelona, Barcelona, Spain. 75 Genk Institute for Fertility Technology, Genk, Belgium. 76 Department of Obstetrics and Gynaecology, ZOL Hospitals and Hasselt University, Genk, Belgium. 77 Department of Oncology and Metabolism, University of Sheffield, Sheffield, UK. 78 Department of Infection, Immunity and Cardiovascular Disease, University of Sheffield, Sheffield, UK. 79 Bloomfield College, Bloomfield, NJ, USA. 80 Department of Urology, Miller School of Medicine, University of Miami, Miami, FL, USA. 81 Division of Reproductive Medicine, Department of Obstetrics and Gynaecologie, Radboud UMC, Nijmegen, The Netherlands. 82 Service Laboratoire de Biologie de la Reproduction-CECOS, Equipe Physiopathologie Surrénalienne et Gonadique, Unité Inserm 1239 NorDic, CHU-Hôpitaux de Rouen, UFR Santé-Université de Rouen, Rouen, France. 83 Department of Biodiversity and Evolutionary Biology, Museo Nacional de Ciencias Naturales (CSIC), Madrid, Spain. 84 Fertility Solutions Inc., Warrensville Heights, OH, USA. 85 Boston IVF, Boston, MA, USA. 86 University Vita-Salute San Raffaele, Milan, Italy. 87 Division of Experimental Oncology/Unit of Urology, URI, IRCCS Ospedale San Raffaele, Milan, Italy. 88 Department of Clinical Chemistry and Molecular Biology, Virgen del Rocío University Hospital, Seville, Spain. 89 Depto de Histologia y Embriología, Facultad de Medicina, Gral. Flores, Uruguay. 90 Centre of Reproductive Medicine and Andrology, Münster, Germany. 91 Department of Urology, Weill Cornell Medicine, New York, NY, USA. 92 Section of Andrology, Department of Urology, Pediatric Urology & Andrology, Justus-Liebig-University/University Hospital of Giessen-Marburg, Giessen, Germany. 93 Lilavati Hospital & Research Centre, Mumbai, India. 94 Human and Animal Physiology, Wageningen University, Wageningen, The Netherlands. 95 WHO Department of Sexual and Reproductive Health and Research (includes the UNDP/UNFPA/UNICEF/WHO/World Bank Special Programme of Research, Development and Research Training in Human Reproduction-HRP), Geneva, Switzerland. 96 Centre for Reproductive Medicine, Vrije Universiteit Brussel, Brussels, Belgium. 97 The Turek Clinic, San Francisco, CA, USA. 98 Medical Bioscience, University of the Western Cape, Bellville, South Africa. 99 Physiology Medical School, Stellenbosch University, Stellenbosch, South Africa. 100 Department of Animal Science, Stellenbosch University, Stellenbosch, South Africa. 101 IBYME, CONICET-FIBYME, Buenos Aires, Argentina. 102 Clinical and Translational Science Institute, The Lundquist Institute, Torrance, CA, USA. 103 Division of Endocrinology, Department of Medicine, Harbor-UCLA Medical Center, Torrance, CA, USA. 104 Fertility Laboratory, Radboud University Medical Centre, Nijmegen, The Netherlands. 105 Thessaloniki Andrology Laboratory-Hellenic Sperm Bank, Thessaloniki, Greece. 106 Laboratory of Histology-Embryology, Medical School, University of Athens, Athens, Greece. 107 Division of Urology, Department of Surgery, St Mary's Hospital, McGill University, Montreal, Canada.

DOI: 10.1093/humrep/deac189
PMID: 36112046
Pubmed: https://pubmed.ncbi.nlm.nih.gov/36112046
Texto completo: https://academic.oup.com/humrep/article-lookup/doi/10.1093/humrep/deac189

Abstract:
Biomedical science is rapidly developing in terms of more transparency, openness and reproducibility of scientific publications. This is even more important for all studies that are based on results from basic semen examination. Recently two concordant documents have been published: the 6th edition of the WHO Laboratory Manual for the Examination and Processing of Human Semen, and the International Standard ISO 23162:2021. With these tools, we propose that authors should be instructed to follow these laboratory methods in order to publish studies in peer-reviewed journals, preferable by using a checklist as suggested in an Appendix to this article.


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