FXTAS is rare among Portuguese patients with movement disorders: FMR1 premutations may be associated with a wider spectrum of phenotypes

The fragile X-associated tremor/ataxia syndrome (FXTAS) is a late-onset neurodegenerative disorder caused by expansions of 55-200 CGG repeats in the 5'UTR of the FMR1 gene. These FMR1 premutation expansions have relatively high frequency in the general population. To estimate the frequency of FMR1 premutations among Portuguese males with non-familial, late-onset movement disorders of unknown etiology, we assessed CGG repeat size in males with disease onset after the age of 50 and negative or unknown family history for late-onset movement disorders, who were sent for SCA, HD, or PD genetic testing at a reference laboratory. The selected patients had a primary clinical diagnosis based on one of the following cardinal features of FXTAS: ataxia, tremor, or cognitive decline. A total of 86 subjects were genotyped for the CGG repeat in the FMR1 gene. We detected one patient with an expansion in the premutation range. The frequency of FMR1 premutations was 1.9% (1/54) in our group of patients with ataxia as the primary clinical feature, and 1.2% (1/86) in the larger movement disorders group. In the family of the FXTAS case, premutation-transmitting females presented a history of psychiatric symptoms, suggesting that, given the wide phenotypical expression of the premutation in females, neuropsychiatric surveillance is necessary. In conclusion, genetic testing for FXTAS should be made available to patients with adult-onset movement disorders to enable adequate genetic counseling to family members

Validation of the rat model of prostate cancer: correlating seminal vesicle lesions with dorsolateral prostate lesions

Background/aim: Lesions in the seminal vesicle are described in the most used protocols for prostate cancer (PCa) induction. This study aimed to characterize the lesions of seminal vesicles associated with a protocol of PCa induction in rats to contribute to better characterization of this model. Materials and methods: Forty-five male Wistar Unilever rats were randomly divided into two control groups: CONT1 (n=10) and CONT2 (n=10); and two PCa-induced groups: IND1 (n=10) and IND2 (n=15), sacrificed at 35 and 61 weeks, respectively. Animals from the induced groups were exposed to a multistep protocol for PCa induction. Animals, seminal vesicles and dorsolateral prostate were weighed. Seminal vesicles and dorsolateral prostate were submitted to histopathological and immunohistochemical analysis. Results: Animals in which PCa was induced had a lower mean body weight when compared with the control animals (p<0.05). The relative mean seminal vesicle weight was higher in groups with PCa when compared with control groups (p<0.05). Although the differences were not statistically significant, animals from the IND2 group developed more lesions than animals from the IND1 and CONT2 groups. It is worth noting that the animals from group IND2 developed papillary adenomas and carcinomas in situ, which were not observed in any other group. Similar to observations in seminal vesicles, animals from group IND2 developed more dorsolateral prostate lesions than animals from the IND1 group (p<0.05). Conclusion: We observed that the longer the exposure to testosterone was, the greater was the incidence of preneoplastic and neoplastic lesions in both the seminal vesicle and the prostate, suggesting that testosterone exposure affects the spectrum of developed lesions

Anatomy and imaging of rat prostate: practical monitoring in experimental cancer-induced protocols

The rat has been frequently used as a model to study several human diseases, including cancer. In many research protocols using cancer models, researchers find it difficult to perform several of the most commonly used techniques and to compare their results. Although the protocols for the study of carcinogenesis are based on the macroscopic and microscopic anatomy of organs, few studies focus on the use of imaging. The use of imaging modalities to monitor the development of cancer avoids the need for intermediate sacrifice to assess the status of induced lesions, thus reducing the number of animals used in experiments. Our work intends to provide a complete and systematic overview of rat prostate anatomy and imaging, facilitating the monitoring of prostate cancer development through different imaging modalities, such as ultrasonography, computed tomography (CT) and magnetic resonance imaging (MRI).publishe

Uloga prirodnih spojeva kod raka mliječnih žlijezda u štakora; blagotvorni učinci vodenog ekstrakta Santolina chamaecyparissus L.

Breast cancer is the most diagnosed cancer among women, and a leading cause of death worldwide. Santolina chamaecyparissus L. is a plant with multiple health benefits, including anticancer and anti-diabetic properties. This study aimed to assess the chemopreventive effects of S. chamaecyparissus aqueous extract (SCE) in an animal model of mammary cancer. A total of 28 four-week-old female Wistar rats were divided into four groups: control, MNU-induced (IND), SCE-supplemented (SCE), and SCE+IND. SCE was added to drinking water (12.72 mg/kg body weight) ad libitum. MNU was administered via the intraperitoneal route at 50 days of age. Weekly monitoring of body weight, food/drink intake, humane endpoints, and number of mammary tumours were recorded. Twenty weeks after MNU administration, animals were sacrificed by anaesthetic overdose and a necropsy was performed. Blood samples were used to determine blood count and serum biochemistry analysis, while kidney and liver samples were analysed for oxidative stress. Tumour samples were collected for gene expression and histology studies. SCE chemical composition was analysed by LC-MS and contained 19 phenolic compounds, with the most abundant being myricetin-O-glucuronide and 1,3-O-dicaffeoylquinic acid. Two animals in the IND group were sacrificed due to exceeding the humane endpoint limits. SCE supplementation delayed mammary tumour development, reducing its volume and weight. SCE had a positive impact on haematological parameters, particularly the neutrophil-lymphocyte ratio (P=0.026). No significant differences were observed in serum biochemistry, except for creatinine kinase MB, or in oxidative stress markers. Gene expression analysis showed significantly reduced VEGF expression levels (P=0.0158) in tumours from SCE+IND. These findings suggest that SCE is deserving of further study to identify the individual compounds and to understand its influence on animal models during cancer development.Rak dojke najčešće je dijagnosticiran rak u žena i vodeći uzrok smrti na svijetu. Santolina chamaecyparissus L. je biljka s višestrukim blagotvornim učincima za zdravlje, uključujući antitumorska i antidijabetička svojstva. Cilj je ove studije bio procijeniti kemopreventivne učinke vodenog ekstrakta S. chamaecyparissus (SCE) na životinjama obeljelim od raka mliječnih žlijezda. Dvadeset i osam četiri ženki starih tjedna wistar štakora podijeljeno je u četiri skupine: kontrolnu, MNU-inducirano (IND), s dodatkom SCE (SCE) i SCE+IND. Skupini SCE je dodan vodi za piće (12,72 mg/kg tjelesne mase) ad libitum; MNU je primijenjen intraperitonealnim putem u 50. danu života. Tjedno je bilježeno praćenje tjelesne mase, unosa hrane/tekućine, humano usmrćivanje i broj tumora mliječnih žlijezda. Dvadeset tjedana nakon primjene MNU, životinje su žrtvovane predoziranjem anestetikom i obavljena je razudba. Uzorci krvi su rabljeni za određivanje krvne slike i analizu biokemije seruma, dok su uzeti uzorci bubrega i pluća rabljeni za analize oksidativnog stresa. Uzorci tumora su prikupljeni za studije ekspresije gena i histološke studije. Analiziran je kemijski sastav skupine SCE pomoću LC-MS i otkriveno je da sadrži 19 fenolnih spojeva od kojih su najobilniji bili miricetin-O-glukuronid i 1,3-O-dikafeoilkina kiselina. Dvije životinje iz IND skupini žrtvovane su zbog prekoračenja ograničenja za humano usmrćivanje. Skupini SCE dodatak je odgodio razvoj tumora mliječnih žlijezda, smanjujući njegov volumen i masu. Skupina SCE je imala pozitivni učinak na hematološke parametre, posebice na omjer neutrofila i limfocita (P=0,026). Nikakve značajne razlike nisu otkrivene u biokemiji seruma, osim kreatinin kinaze MB, niti u markerima oksidativnog stresa. Analiza ekspresije gena pokazala je značajno smanjene razina ekspresije VEGF (P=0,0158) u tumora iz skupine SCE+IND. Ovi nalazi ukazuju da bi skupinu SCE trebalo dodatno ispitati da bi se identificirali pojedinačni spojevi i razumio njegov utjecaj na životinjama oboljelih od raka mliječnih žlijezda

Crecimiento de rotiferos con Nannochloropsis gaditana cultivada en fotobiorreactores versus cultivo tradicional en bolsas

En las instalaciones de la planta de cultivos marinos del Centro Oceanoqráñco de Vigo (lEO) se cuHivan diversas especies de microaJgas seg1ÍI! sistema tradicional de producción en bolsas (5OOL) y, mayoritariamente; bajo un nuevo sistema de producción en lotobiorreac!ores (FBRs)de columna (1 ooUUd.) de ano rendimiento montados por la empresa AQUALGAE SL. Parte de la producción de microalgas se destina al OJltivoy enriquecimiento de alimento vivo (rotíferos y Artemia). Este experimento se diseñó con el fin de obtener resutados que nos permitan asegurara calidad del cultivo de rotífero, aportando como alimento las microalgas producidas en semicontinuoen los FBRs, para poder así sustituirtotalmenle el sistema tradicional de producción en bolsas, menos productivo, más laborioso y sin apenas control sobre la composición nutricional dela biomasa cosechada. Los resultados obtenidos demuestran la calidad y estabilidad nutricional de las microalgas producidas en los FBRs,11) habiendo dilerencias signilicativas en el crecimiento de las poblaciones de rotíferos, aunque el número total de huevos sí ha sido superior en ~ grupo alimentado con microalgas procedentes de los FBRs. Se discuten además las dilerencias se composición nutricional observadas.In lhe lacilijies 01 lhe Centro Oceanográfico de Vigo (lEO) several species 01 microalgae are produced according to the traditional batch OJltureil poIyelhylene bags (500 L) ando more recently, in high yield column photobiorreactors (PBRs), 100 L each column, installed by tIle ~ AQUALGAE SL. Part 01 lhe harvested microalgae biomass is used lor lhe culture and enrichment 01 live prey (rotiler and Artemia). The goal cíl!e lhis wor1<was to evaluate the leasibilijy 01 maintaining and ensuring a good qualijy 01 rotilers led microalgae cuHured semicontinuously in PBRs,~ orderto lully substiMe tradijional cuHure in bags which are less productive, require more hand wor1<and no control on lhe nutritional compositioncí algae can be exerted. Hesults showed lhe nutritional qualijy 01 lhe microalgae biomass obtained in PBRs, and lhe stability 01 ijs composition. No signilicant differences were observed in comparison with group led algae Irom traditional bags except lor total eggs which were higher in ~ algae lrom PBRs. Nutritional composition differences among group are also discussed

‘Costa da Morte’ ataxia is spinocerebellar ataxia 36: clinical and genetic characterization

Spinocerebellar ataxia 36 has been recently described in Japanese families as a new type of spinocerebellar ataxia with motor neuron signs. It is caused by a GGCCTG repeat expansion in intron 1 of NOP56. Family interview and document research allowed us to reconstruct two extensive, multigenerational kindreds stemming from the same village (Costa da Morte in Galicia, Spain), in the 17th century. We found the presence of the spinocerebellar ataxia 36 mutation co-segregating with disease in these families in whom we had previously identified an ∼0.8 Mb linkage region to chromosome 20 p. Subsequent screening revealed the NOP56 expansion in eight additional Galician ataxia kindreds. While normal alleles contain 5–14 hexanucleotide repeats, expanded alleles range from ∼650 to 2500 repeats, within a shared haplotype. Further expansion of repeat size was frequent, especially upon paternal transmission, while instances of allele contraction were observed in maternal transmissions. We found a total of 63 individuals carrying the mutation, 44 of whom were confirmed to be clinically affected; over 400 people are at risk. We describe here the detailed clinical picture, consisting of a late-onset, slowly progressive cerebellar syndrome with variable eye movement abnormalities and sensorineural hearing loss. There were signs of denervation in the tongue, as well as mild pyramidal signs, but otherwise no signs of classical amyotrophic lateral sclerosis. Magnetic resonance imaging findings were consistent with the clinical course, showing atrophy of the cerebellar vermis in initial stages, later evolving to a pattern of olivo-ponto-cerebellar atrophy. We estimated the origin of the founder mutation in Galicia to have occurred ∼1275 years ago. Out of 160 Galician families with spinocerebellar ataxia, 10 (6.3%) were found to have spinocerebellar ataxia 36, while 15 (9.4%) showed other of the routinely tested dominant spinocerebellar ataxia types. Spinocerebellar ataxia 36 is thus, so far, the most frequent dominant spinocerebellar ataxia in this region, which may have implications for American countries associated with traditional Spanish emigration

Immune and spermatogenesis-related loci are involved in the development of extreme patterns of male infertility

We conducted a genome-wide association study in a large population of infertile men due to unexplained spermatogenic failure (SPGF). More than seven million genetic variants were analysed in 1,274 SPGF cases and 1,951 unaffected controls from two independent European cohorts. Two genomic regions were associated with the most severe histological pattern of SPGF, defined by Sertoli cell-only (SCO) phenotype, namely the MHC class II gene HLA-DRB1 (rs1136759, P = 1.32E-08, OR = 1.80) and an upstream locus of VRK1 (rs115054029, P = 4.24E-08, OR = 3.14), which encodes a protein kinase involved in the regulation of spermatogenesis. The SCO-associated rs1136759 allele (G) determines a serine in the position 13 of the HLA-DR beta 1 molecule located in the antigen-binding pocket. Overall, our data support the notion of unexplained SPGF as a complex trait influenced by common variation in the genome, with the SCO phenotype likely representing an immune-mediated condition. A GWAS in a large case-control cohort of European ancestry identifies two genomic regions, the MHC class II gene HLA-DRB1 and an upstream locus of VRK1, that are associated with the most severe phenotype of spermatogenic failure

Evaluation of male fertility-associated loci in a european population of patients with severe spermatogenic impairment

Funding: This work was supported by the Spanish Ministry of Economy and Competitiveness through the Spanish State Plan for Scientific and Technical Research and Innovation (ref. SAF2016-78722-R), the “Ramón y Cajal” program (ref. RYC-2014-16458), and the “Juan de la Cierva Incorporación” program (ref. IJC2018-038026-I), which include FEDER funds. SLa received support from the Spanish Ministry of Science and Innovation (grants FIS-ISCIII DTS18/00101, co-funded by FEDER funds/European Regional Development Fund (ERDF)-a way to build Europe-), and from Generalitat de Catalunya (grant 2017SGR191). AG-J was recipient of a grant from the “Plan Propio” program of the University of Granada (“Becas de Iniciación a la Investigación para estudiantes de Grado”, conv.2019). SLa is sponsored by the “Researchers Consolidation Program” from the SNS-Dpt. Salut Generalitat de Catalunya (Exp. CES09/020). JG was partially funded by FCT/MCTES, through national funds attributed to Center for Toxicogenomics and Human Health—ToxOmics (UIDB/00009/2020). PIM is supported by the FCT post-doctoral fellowship (SFRH/BPD/120777/2016), financed from the Portuguese State Budget of the Ministry for Science, Technology and High Education and from the European Social Fund, available through the Programa Operacional do Capital Humano. AML is funded by the Portuguese Government through FCT (IF/01262/2014). IPATIMUP integrates the i3S Research Unit, which is partially supported by FCT in the framework of the project “Institute for Research and Innovation in Health Sciences” (POCI-01-0145-FEDER-007274).Infertility is a growing concern in developed societies. Two extreme phenotypes of male infertility are non-obstructive azoospermia (NOA) and severe oligospermia (SO), which are characterized by severe spermatogenic failure (SpF). We designed a genetic association study comprising 725 Iberian infertile men as a consequence of SpF and 1058 unaffected controls to evaluate whether five single-nucleotide polymorphisms (SNPs), previously associated with reduced fertility in Hutterites, are also involved in the genetic susceptibility to idiopathic SpF and specific clinical entities. A significant difference in the allele frequencies of USP8-rs7174015 was observed under the recessive model between the NOA group and both the control group (p = 0.0226, OR = 1.33) and the SO group (p = 0.0048, OR = 1.78). Other genetic associations for EPSTI1-rs12870438 and PSAT1-rs7867029 with SO and between TUSC1-rs10966811 and testicular sperm extraction (TESE) success in the context of NOA were observed. In silico analysis of functional annotations demonstrated cis-eQTL effects of such SNPs likely due to the modification of binding motif sites for relevant transcription factors of the spermatogenic process. The findings reported here shed light on the molecular mechanisms leading to severe phenotypes of idiopathic male infertility, and may help to better understand the contribution of the common genetic variation to the development of these conditions.publishersversionpublishe