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1.
Sci Total Environ ; 879: 163040, 2023 Jun 25.
Article in English | MEDLINE | ID: mdl-36965720

ABSTRACT

The loggerhead sea turtle (Caretta caretta) has been selected as sentinel species by the Marine Strategy Framework Directive (MSFD) descriptor 10 in relation to marine litter. In this, and other protected species, there is a need to develop conservative pollution biomarkers equally informative of chemical exposures to those traditionally carried out in metabolic organs, such as the liver. With this aim, plasma from turtles undergoing rehabilitation at the Fundació Oceanogràfic rescue centre (Arca del Mar) were selected and tested for B-esterase measurements. Hydrolysis rates of acetylcholinesterase (AChE), butyrylcholinesterase (BChE) and carboxylesterases (CEs) using four commercial substrates were undertaken on 191 plasma samples. Results indicated that acetylthiocholine was the most adequate substrate of cholinesterases and butyrate esters for CE measures. The correlation of these parameters with well-established blood biochemistry measurements was analysed. B-esterase measures in wild specimens were discussed in relation to age group, pathology on admission to the rescue centre and season; moreover, contrasts with long-term resident turtles were also made. Although this study provides baseline data on B-esterase measures in a large sample size for this species, more complementary information is still needed in terms of population genetics, chemical exposures, and in relation to other biochemical parameters before they can be confidently applied in wild specimens within the regulatory MSFD.


Subject(s)
Turtles , Animals , Carboxylesterase/metabolism , Acetylcholinesterase/metabolism , Butyrylcholinesterase/metabolism , Health Status
2.
Rev. esp. med. nucl. imagen mol. (Ed. impr.) ; 41(5): 292-299, sept. - oct. 2022. ilus, tab
Article in Spanish | IBECS | ID: ibc-211036

ABSTRACT

Objetivo Estudiar la viabilidad y utilidad de la técnica combinada biopsia selectiva de ganglio centinela (BSGC)-biopsia radioguiada del ganglio patológico en pacientes con cáncer de mama y afectación axilar sometidas a quimioterapia neoadyuvante (QTNA). aterial y métodos Estudio prospectivo en el que se incluyeron 30 pacientes con cáncer de mama y estadificación axilar cN1, con indicación de QTNA. El ganglio biopsiado se marcó (GBM) mediante un clip, antes del tratamiento QTNA. Tras completar este se valoró la respuesta axilar con ecografía. En caso de buena respuesta ecográfica se procedió a realizar BSGC (99mTc-nanocoloides) y disección axilar dirigida mediante biopsia radioguiada del GBM (99mTc-macroagregados albúmina). Se procedía a realizar una linfadenectomía axilar si el ganglio centinela (GC) y/o el GBM contenían células tumorales. Se evaluaron las tasas de localización-exéresis del GC y del GBM. También se evaluaron la tasa de falsos negativos y el valor predictivo positivo de la BSGC sola. Resultados Se extirpó el GC en todas las pacientes, mientras que el GBM se extirpó exitosamente en 27 (90%). El GC coincidió con el GBM en 15 pacientes (50%). En 12 pacientes el GC fue negativo pero el GBM fue positivo para metástasis, lo que comportó una tasa de falsos negativos del 44,4% y un valor predictivo positivo del 37% para la BSGC sola. Tanto el GC como el GBM resultaron negativos en 5 pacientes (18,5%), lo que permitió evitar realizarles la linfadenectomía axilar. Conclusiones La técnica combinada descrita BSGC-biopsia radioguiada del ganglio patológico es un procedimiento útil y accesible para la reestadificación precisa de la axila tras QTNA, evitando la alta tasa de falsos negativos de la BSGC sola en este grupo de pacientes y evitando un número mayor de linfadenectomías axilares (AU)


Objective To study the feasibility and usefulness of the combined technique selective sentinel lymph node biopsy (SLNB)-radioguided biopsy of the pathological lymph node in patients with breast cancer and axillary involvement undergoing neoadjuvant chemotherapy (NAC). Material and methods Prospective diagnostic study of 30 patients with breast cancer and cN1 axillary staging with NACT indication. Before NACT, the biopsied node was marked with a clip (MBN). After NACT an ultrasound was performed and in case of good response a SLNB (99mTc-nanocolloids) plus targeted axillary dissection MBN pathologic node radioguided biopsy (99mTc-albumin macroaggregates) was performed. Axillary lymph node dissection was performed if SLNB and/or MBN were positive for tumor cells. The localization-removal rate of the sentinel lymph node (SLN) and MBN were evaluated. False-negative rate and positive predictive value of SLNB alone were also evaluated. Results Thirty patients were included in the study. SLN could be detected in all patients while MBN was successfully removed in 27 (90%). The SLN coincided with MBN in 15 patients (50%). In 12 patients SLNB was negative while MBN positive, leading to a FNR of 44,4% for SLNB alone. We found a positive predictive value of 37% for the SLNB. In 5 patients (18,5%) both SLNB and MBN were negative, avoiding axillary lymph node dissection. Conclusions SLNB-MBN radioguided biopsy combined technique is a useful and accessible procedure for accurate axillary restaging after NACT, avoiding the high rate of false-negative rate of SLNB alone in this group of patients and avoiding a great number of axillary lymph node dissection (AU)


Subject(s)
Humans , Female , Adult , Middle Aged , Aged , Breast Neoplasms/drug therapy , Breast Neoplasms/surgery , Breast Neoplasms/diagnostic imaging , Lymphatic Metastasis/diagnostic imaging , Neoadjuvant Therapy , Prospective Studies , Neoplasm Staging
3.
Environ Pollut ; 313: 120149, 2022 Nov 15.
Article in English | MEDLINE | ID: mdl-36115493

ABSTRACT

A total of 164 blood samples from 16 clinically healthy bottlenose dolphins (Tursiops truncatus), were obtained from an aquarium in Spain between 2019 and 2020, as part of their preventive medicine protocol. In addition to conventional haematological and biochemical analyses, plasmatic B-esterase activities were characterised to determine the potential application of such analyses in wild counterparts. The hydrolysis rates for the substrates of acetylcholinesterase (AChE), butyrylcholinesterase (BuChE) and carboxylesterase (CE) activity in plasma were measured, the last using two commercial substrates, p-nitrophenyl acetate (pNPA) and p-nitrophenyl butyrate (pNPB). Activity rates (mean ± SEM in nmol/min/mL plasma) were (in descending order): AChE (125.6 ± 3.8), pNPB-CE (65.0 ± 2.2), pNPA-CE (49.7 ± 1.1) and BuChE (12.8 ± 1.3). These values for dolphins are reported in here for the first time in this species. Additionally, the in vitro sensitivity of two B-esterases (AChE and pNPB-CE) to chemicals of environmental concern was determined, and the protective role of plasmatic albumin assessed. Out of the B-esterases measured in plasma of dolphin, AChE activity was more responsive in vitro to pesticides, while CEs had a low response to plastic additives, likely due to the protective presence of albumin. However, the clear in vitro interaction of these environmental chemicals with purified AChE from electric eels and recombinant human hCEs (hCE1 and hCE2) and albumin, predicts their impact in other tissues that require in vivo validation. A relationship between esterase-like activities and health parameters in terrestrial mammals has already been established. Thus, B-esterase measures could be easily included in marine mammal health assessment protocols for dolphins as well, once the relationship between these measures and the animal's fitness has been established.


Subject(s)
Bottle-Nosed Dolphin , Pesticides , Acetylcholinesterase , Albumins , Animals , Biomarkers , Butyrylcholinesterase , Carboxylesterase , Esterases , Humans , Nitrophenols , Plastics , Xenobiotics
4.
Chromosoma ; 131(4): 193-205, 2022 12.
Article in English | MEDLINE | ID: mdl-35960388

ABSTRACT

In most eukaryotes, pairing of homologous chromosomes is an essential feature of meiosis that ensures homologous recombination and segregation. However, when the pairing process begins, it is still under investigation. Contrasting data exists in Mus musculus, since both leptotene DSB-dependent and preleptotene DSB-independent mechanisms have been described. To unravel this contention, we examined homologous pairing in pre-meiotic and meiotic Mus musculus cells using a three-dimensional fluorescence in situ hybridization-based protocol, which enables the analysis of the entire karyotype using DNA painting probes. Our data establishes in an unambiguously manner that 73.83% of homologous chromosomes are already paired at premeiotic stages (spermatogonia-early preleptotene spermatocytes). The percentage of paired homologous chromosomes increases to 84.60% at mid-preleptotene-zygotene stage, reaching 100% at pachytene stage. Importantly, our results demonstrate a high percentage of homologous pairing observed before the onset of meiosis; this pairing does not occur randomly, as the percentage was higher than that observed in somatic cells (19.47%) and between nonhomologous chromosomes (41.1%). Finally, we have also observed that premeiotic homologous pairing is asynchronous and independent of the chromosome size, GC content, or presence of NOR regions.


Subject(s)
Meiosis , Meiotic Prophase I , Animals , Mice , Male , In Situ Hybridization, Fluorescence , Karyotype , Spermatocytes
5.
Environ Res ; 213: 113639, 2022 10.
Article in English | MEDLINE | ID: mdl-35688215

ABSTRACT

Sea turtles are particularly vulnerable to plastic exposures, and the associated chemical additives, due to their feeding strategies. The species Caretta caretta is a proposed sentinel of plastic pollution worldwide. Thus, there is a need to find adequate biomarkers of plastic exposure through non-invasive protocols for this IUCN protected species. Plasmatic acetylcholinesterase (AChE), butyrylcholinesterase (BuChE) and carboxylesterase (CE) which participate in xenobiotic and endogenous metabolic reactions could all serve as biomarkers, as they are responsive to plasticizers and have already proved adequate for identifying organophosphorus esters exposures. Here we measured plasmatic B-esterases in wild specimens captured as accidental by-catch. Measurements were taken in each individual either at entry into the rehabilitation program or immediately before release after a recovery period. For CE measurements, 4 commercial substrates were used as potentially indicative of distinct enzyme isoforms. Increased activity was seen with the butyrate-derived substrates. Plasmatic CE activities were over one order of magnitude higher than AChE and BuChE substrates. Moreover, an in vitro protocol with the inclusion of plastic additives such as tetrabromobisphenol A (TBBPA), bisphenol A and some of its analogues was considered a proxy of enzymatic interactions. A clear inhibition by TBBPA was found when using commercially purified AChE and recombinant CE proteins. Overall, from in vitro and in vivo evidences, CEs in plasma are sensitive and easily measurable and have been shown to significantly increase after turtles have been rehabilitated in rescue centres. Nevertheless, the inclusion of plastic (or plasticizers) characterisation would help to confirm its association with plasmatic enzyme modifications before they can be adopted as biomarkers of plastic contamination.


Subject(s)
Turtles , Water Pollutants , Acetylcholinesterase , Animals , Biomarkers , Butyrylcholinesterase , Esterases , Plasticizers , Plastics , Water Pollutants/analysis
6.
Article in English | MEDLINE | ID: mdl-35597764

ABSTRACT

OBJECTIVE: To study the feasibility and usefulness of ultrasound-guided pre-chemotherapy marking of pathologic lymph node followed by sentinel lymph node biopsy (SLNB)-pathologic node radioguided biopsy (ROLL) combined technique, in axillary involvement breast cancer patients undergoing neoadjuvant chemotherapy (NACT). MATERIAL AND METHODS: Prospective diagnostic study of 30 patients with breast cancer and cN1 axillary staging with NACT indication. Before NACT, the biopsied node was marked with a clip (MBN). After NACT an ultrasound was performed and in case of good response a SLNB (99mTc-nanocolloids) plus targeted axillary dissection MBN ROLL biopsy (99mTc-albumin macroaggregates) was performed. Axillary lymph node dissection (ALND) was performed if SLNB and/or MBN were positive for tumor cells. The localization-removal rate of the sentinel lymph node (SLN) and MBN were evaluated. False-negative rate (FNR) and positive predictive value (PPV) of SLNB alone were also evaluated. RESULTS: Thirty patients were included in the study. SLN could be detected in all patients while MBN was successfully removed in 27 (90%). The SLN coincided with MBN in 15 patients (50%). In 12 patients SLNB was negative while MBN positive, leading to a FNR of 44.4% for SLNB alone. We found a PPV of 37% for the SLNB. In 5 patients (18.5%) both SLNB and MBN were negative, avoiding ALND. CONCLUSIONS: SLNB-MBN radioguided biopsy ROLL combined technique is a useful and accessible procedure for accurate axillary restaging after NACT, avoiding the high rate of FNR of SLNB alone in this group of patients and avoiding a great number of ALND.


Subject(s)
Breast Neoplasms , Sentinel Lymph Node , Surgery, Computer-Assisted , Breast Neoplasms/diagnostic imaging , Breast Neoplasms/drug therapy , Breast Neoplasms/surgery , Female , Humans , Lymphatic Metastasis/diagnostic imaging , Neoadjuvant Therapy/methods , Prospective Studies , Sentinel Lymph Node/diagnostic imaging , Sentinel Lymph Node/pathology , Sentinel Lymph Node/surgery , Sentinel Lymph Node Biopsy/methods
7.
Exp Eye Res ; 218: 109026, 2022 05.
Article in English | MEDLINE | ID: mdl-35276184

ABSTRACT

Although many more eyes of children with retinoblastoma are salvaged now compared to just 10 years ago, the control of vitreous seeding remains a challenge. The introduction of intravitreal injection of melphalan has enabled more eyes to be salvaged safely but with definite retinal toxicity. Intensive treatment with high-dose intravitreal topotecan may be a strategy to control tumor burden because of its cell cycle-dependent cytotoxicity and the proven safety in humans. Therefore, we evaluated the ocular and systemic safety of repeated high-dose intravitreal injections of topotecan in rabbits. Systemic and ocular toxicity was assessed in non-tumor-bearing rabbits after four weekly injections of three doses of topotecan (10 µg, 25 µg, and 50 µg) or vehicle alone. Animals were evaluated weekly for general and ophthalmic clinical status. One week after the last injection, vitreous and plasma samples were collected for drug quantification and the enucleated eyes were subjected to histological assessment. Weight, hair loss, or changes in hematologic values were absent during the study period across all animal groups. Eyes injected with all topotecan doses or vehicle showed no signs of anterior segment inflammation, clinical or histologic evidence of damage to the retina, and ERG parameters remained unaltered throughout the study. Vitreous and plasma topotecan lactone concentrations were undetectable. Four weekly intravitreal injections of topotecan up to 50 µg in the animal model or a 100 µg human equivalent dose were not toxic for the rabbit eye. High doses of topotecan may show promising translation to the clinic for the management of difficult-to-treat retinoblastoma vitreous seeds.


Subject(s)
Retinal Neoplasms , Retinoblastoma , Animals , Intravitreal Injections , Rabbits , Retinal Neoplasms/drug therapy , Retinal Neoplasms/pathology , Retinoblastoma/drug therapy , Retinoblastoma/pathology , Topotecan/toxicity , Vitreous Body/pathology
8.
Environ Res ; 210: 112961, 2022 07.
Article in English | MEDLINE | ID: mdl-35181305

ABSTRACT

Cephalopods are a group of marine invertebrates that have received little attention as sentinel species in comparison to other molluscs, such as bivalves. Consequently, their physiological and biochemical xenobiotic metabolism responses are poorly understood. Here we undertake a comparative analysis of the enzymatic activities involved in detoxification reactions and neural transmission in the digestive tract of two commercial cephalopods: the Common octopus, Octopus vulgaris, and the European cuttlefish, Sepia officinalis. For methodological purposes, several common B-esterases (five carboxylesterase (CE) substrates and three cholinesterase (ChE) determinations) were assayed as a proxy of metabolic and neuronal activities, respectively. Four components of the digestive tract in each species were considered: salivary glands, the stomach, the digestive gland and the caecum. The in vitro responses of digestive gland homogenates to model chemicals and contaminants of environmental concern were contrasted between both cephalopod species. The baseline biochemical activities in the four digestive tract components were also determined. Moreover, in order to validate the protocol, purified proteins, recombinant human CE (CE1 and CE2) and purified eel acetylcholinesterase (AChE) were included in the analysis. Overall, carboxylesterase activities were higher in octopus than in cuttlefish, with the activity quantified in the digestive tract components in the following order: digestive gland ≈ caecum > stomach ≈ salivary glands, with higher hydrolysis rates reached with naphthyl-derived substrates. In contrast, cuttlefish hydrolysis rates with ChE substrates were higher than in octopus. This trend was also reflected in a higher sensitivity to CE inhibitors in octopus and to AChE inhibitors in cuttlefish. Given the detoxification character of CEs and its protective role preventing AChE inhibition, octopus could be regarded as more efficiently protected than cuttlefish from neurotoxic exposures. A full characterisation of B-esterases in the digestive tract of the two common cephalopods is also provided.


Subject(s)
Carboxylesterase , Octopodiformes , Acetylcholinesterase/metabolism , Animals , Carboxylesterase/metabolism , Cholinesterases/metabolism , Decapodiformes/metabolism , Gastrointestinal Tract/metabolism , Humans , Octopodiformes/metabolism
9.
Article in Spanish | IBECS | ID: ibc-211470

ABSTRACT

Las vías de conocimiento de los practicantes hasta tiempos contemporáneos siempre han sido precarias, sobre todo, en las zonas rurales y de montaña, donde la información llegaba a cuentagotas. La comunicación sufrió una pequeña revolución a principios del siglo XX con la aparición de diversas publicaciones en forma de revistas. Algunas de ellas eran de alcance estatal, pero, poco a poco, la conciencia de clase los llevó a intentar acometer empresas de mayor envergadura en forma de boletines que representaran los intereses y las reivindicaciones de los emergentes y noveles colegios provinciales [Fragmento de texto] (AU)


Subject(s)
Humans , History, 20th Century , Periodicals as Topic/history , Scientific Communication and Diffusion , Spain
10.
Materials (Basel) ; 14(19)2021 Oct 03.
Article in English | MEDLINE | ID: mdl-34640192

ABSTRACT

Olive mill wastes represent an important environmental problem. Their high phenol, lipid, and organic acid concentrations turn them into phytotoxic materials. Specifically, wet olive pomace (WOP) is the waste generated in the two-phase continuous extraction process. WOP is a paste with around 60% water. The total volume of WOP generated is around 0.25 L/kg of olives processed. Its current waste management practices result in environmental problems as soil contamination, underground seepage, water-bodies pollution, and foul odor emissions. Some valorization alternatives include composting, biological treatments, direct combustion for energy production, or direct land application. The leather industry is making great efforts to apply cleaner processes while substituting chemical products for natural products. In this way, different alternatives are being studied, such as the use of zeolites, triazine derivatives, grape seed extract, olive leaf extract, etc. In this work, the use of wet olive pomace is presented as a possible alternative to conventional vegetable tannins (mimosa, quebracho, chestnut, etc.). Although different projects and studies have been developed for the valorization of olive mill wastes, there is completely a new approach to the WOP application for tanning purposes. This study shows that WOP has a significant number of polyphenolic substances, so it has a great potential to be used as a tanning agent. Specifically, this study has been able to determine that, of the polyphenols present in WOP, 39.6% correspond to tannins that are capable of tanning the skin. Additionally, it contains 14.3% non-tannins, that is, molecules that by themselves do not have the capacity to tan the leather but promote the tanning mechanism and improve the properties of the tanned leather.

11.
Chromosoma ; 130(2-3): 163-175, 2021 09.
Article in English | MEDLINE | ID: mdl-34231035

ABSTRACT

Chromosome territoriality is not random along the cell cycle and it is mainly governed by intrinsic chromosome factors and gene expression patterns. Conversely, very few studies have explored the factors that determine chromosome territoriality and its influencing factors during meiosis. In this study, we analysed chromosome positioning in murine spermatogenic cells using three-dimensionally fluorescence in situ hybridization-based methodology, which allows the analysis of the entire karyotype. The main objective of the study was to decipher chromosome positioning in a radial axis (all analysed germ-cell nuclei) and longitudinal axis (only spermatozoa) and to identify the chromosomal factors that regulate such an arrangement. Results demonstrated that the radial positioning of chromosomes during spermatogenesis was cell-type specific and influenced by chromosomal factors associated to gene activity. Chromosomes with specific features that enhance transcription (high GC content, high gene density and high numbers of predicted expressed genes) were preferentially observed in the inner part of the nucleus in virtually all cell types. Moreover, the position of the sex chromosomes was influenced by their transcriptional status, from the periphery of the nucleus when its activity was repressed (pachytene) to a more internal position when it is partially activated (spermatid). At pachytene, chromosome positioning was also influenced by chromosome size due to the bouquet formation. Longitudinal chromosome positioning in the sperm nucleus was not random either, suggesting the importance of ordered longitudinal positioning for the release and activation of the paternal genome after fertilisation.


Subject(s)
Chromosome Positioning , Meiosis , Animals , In Situ Hybridization, Fluorescence , Male , Mice , Sex Chromosomes , Spermatogenesis/genetics
12.
Abdom Radiol (NY) ; 46(7): 3280-3287, 2021 07.
Article in English | MEDLINE | ID: mdl-33674961

ABSTRACT

PURPOSE: To identify specific contrast-enhanced CT (CECT) findings and develop a predictive model with logistic regression to differentiate fat-poor angiomyolipomas (fpAML) from papillary renal cell carcinomas (pRCC). METHODS: This is a single-institution retrospective study that assess CT features of histologically proven 67 pRCC and 13 fpAML. CECT variables were studied by means of univariate logistic regression. Variables included patients' demographics, tumor attenuation (unenhanced and at arterial, venous and excretory post-contrast phases), type of enhancement, morphological features (axial long and short diameters, long-short axis ratio (LSR) and tumor to kidney angle interface) and presence of visible calcifications or vessels. Those variables with a p ≤ 0.05 underwent standard stepwise logistic regression to find predictive combinations of clinical variables. Best models were evaluated by AUROC curves and were subjected to Leave-one-out cross validation to assess their robustness. RESULTS: Odds ratio (OR) between pRCC and fpAML was statistically significant for patient's gender, tumor attenuation in arterial, venous and excretory phases, tumor's long diameter, short diameter, LSR, type of enhancement, presence of intratumoral vessels and tumor-kidney angle interface. The best predictive model resulted in an area under the curve (AUC) of 0.971 and included gender, tumor-kidney angle interface and venous attenuation with the following equation: Log(p/1 - p) = - 2.834 + 4.052 * gender + - 0.066 * AngleInterface + 0.074 * VenousphaseHU. CONCLUSIONS: The combination of patients' gender, tumor to kidney angle interface and venous enhancement helps to distinguish fpAML from pRCC.


Subject(s)
Angiomyolipoma , Carcinoma, Renal Cell , Kidney Neoplasms , Angiomyolipoma/diagnostic imaging , Carcinoma, Renal Cell/diagnostic imaging , Cell Differentiation , Contrast Media , Diagnosis, Differential , Humans , Kidney Neoplasms/diagnostic imaging , Retrospective Studies , Sensitivity and Specificity , Tomography, X-Ray Computed
13.
Chemosphere ; 277: 130283, 2021 Aug.
Article in English | MEDLINE | ID: mdl-33774234

ABSTRACT

The earthworm Lumbricus terrestris is an anecic species living in natural soils but it is also a sentinel in pollution monitoring. Specimens of L.terrestris were exposed for 48 h though the filter paper contact test at 1 mg/mL of the chemicals: Lamotrigine (LMG), Cocaine (COC), Fipronil (FIP) and the pesticide bis-4-nitrophenyl phosphate (BNPP). After that period, the activities of Acetylcholinesterase, Glutathione S-transferase, Carboxylesterase (CE) using different substrates, and lipid peroxidation levels were evaluated in the exposed whole tissue earthworms. The results revealed differences only in CE activity, with 4-nitrophenyl butyrate (4NPB) and 1-naphthyl butyrate (1NB) the most responsive substrates to COC. The kinetic parameters of CE were characterized, for the first time, in whole tissue of this species. The chemical analysis by LC-MS/MS, confirmed the exposure to the parent compounds, identified metabolites and evidenced biotransformation pathways in earthworms. Metabolic reactions included oxidation (LMG and FIP), hydrolysis (COC and FIP) as well as glycosylation (LMG, COC and FIP). A hitherto unknown metabolite of LMG due to the conjugation with phenylalanine glutamine was formed. The in vivo results on CE activity with the specific inhibitor, BNPP, were confirmed in vitro. Moreover, in the in vitro approach, the inclusion of other contaminants of environmental concern supports the potential of CE as biomarker. This study identifies the main metabolites formed by earthworms for further in vivo exposures under more realistic conditions and the potential use of CE measures as biomarker of emerging contaminants.


Subject(s)
Oligochaeta , Pharmaceutical Preparations , Soil Pollutants , Animals , Biomarkers , Chromatography, Liquid , Soil Pollutants/toxicity , Tandem Mass Spectrometry
14.
J Nephrol ; 34(3): 689-697, 2021 06.
Article in English | MEDLINE | ID: mdl-32876939

ABSTRACT

BACKGROUND: Ex vivo confocal microscopy is a technique for tissue examination, which generates images of fresh samples with an optical resolution comparable to those obtained by conventional pathology. The objective of this study was to evaluate the feasibility of using ex vivo confocal microscopy in fusion mode (reflectance and fluorescence) and the H&E-like digital staining that is obtained for the analysis of non-neoplastic kidney biopsies. METHODS: Twenty-four renal samples acquired from autopsies were scanned in a 4th generation ex vivo confocal microscopy device. The imaging process was completed in an average of three minutes. RESULTS: Confocal images correlated very well to the corresponding conventional histological sections, both in normal tissue and in chronic lesions (glomerulosclerosis, fibrosis and tubular atrophy). The ex vivo confocal microscopy protocol did not add artifacts to the sample for the ulterior study with light microscopy, nor to the histochemical or immunohistochemical studies. CONCLUSION: The ease and speed of grayscale and fluorescence image acquisition, together with the quality of the H&E-like digitally stained images obtained with this approach, suggest that this technique shows promise for use in clinical nephrology and renal transplantation.


Subject(s)
Microscopy, Confocal , Biopsy , Humans , Staining and Labeling
15.
Equine Vet J ; 52(2): 174-180, 2020 Mar.
Article in English | MEDLINE | ID: mdl-31461557

ABSTRACT

BACKGROUND: Dynamic laryngeal collapse (DLC) associated with poll flexion is the most common disorder of the upper respiratory tract (URT) in the Norwegian-Swedish Coldblooded Trotter (NSCT). The disorder, which has also been diagnosed in other breeds of trotters and gaited horses, appears to be related to anatomic phenotypes and only occurs during poll flexion when the horse is exercised 'on the bit'. OBJECTIVES: Identify genomic regions associated with DLC in the NSCT by combining a rigorous phenotyping protocol with genomic data from a high-density equine genotyping array. STUDY DESIGN: Prospective case/control study. METHODS: High-speed treadmill endoscopy was used to phenotype horses (n = 61) for DLC, distinguishing between cases and controls. Genome-wide association (GWA) analysis of DLC status was then performed using a principal component approach (PCA) with haplotype analyses subsequently performed for regions containing single-nucleotide polymorphisms (SNPs) above the suggestive genome-wide significance (GWS) threshold (P<1.0 × 10-5 ). RESULTS: One region containing 10 SNPs (Equus caballus chromosome [ECA] 7: 89,601,935-94,647,192) was above the suggestive GWS threshold. Two inferred haplotypes in this region demonstrated significant differences (P<0.001) between cases and controls, with the most frequent haplotype resulting in a significantly increased risk of DLC. MAIN LIMITATIONS: Small sample size as a result of stringent phenotyping protocols. CONCLUSIONS: The current study highlights a candidate genomic region on ECA7 as potentially important with regard to the manifestation of DLC. Further exploration of this region and the genes included within it will bring veterinarians and researchers closer to fully understanding the biological mechanisms underlying DLC in horses.


Subject(s)
Horse Diseases , Larynx , Animals , Genome-Wide Association Study/veterinary , Horses , Norway , Prospective Studies , Sweden
16.
Sci Rep ; 9(1): 9340, 2019 06 27.
Article in English | MEDLINE | ID: mdl-31249355

ABSTRACT

Recent studies, both in laboratory and sea conditions, have demonstrated damage after sound exposure in the cephalopod statocyst sensory epithelium, which secretes endolymph protein. Here, the proteomic analysis of the endolymph was performed before and after sound exposure to assess the effects of exposure to low intensity, low frequency sounds on the statocyst endolymph of the Mediterranean common cuttlefish (Sepia officinalis), determining changes in the protein composition of the statocyst endolymph immediately and 24 h after sound exposure. Significant differences in protein expression were observed, especially 24 h after exposure. A total of 37 spots were significantly different in exposed specimens, 17 of which were mostly related to stress and cytoskeletal structure. Among the stress proteins eight spots corresponding to eight hemocyanin isoforms were under-expressed possible due to lower oxygen consumption. In addition, cytoskeletal proteins such as tubulin alpha chain and intermediate filament protein were also down-regulated after exposure. Thus, endolymph analysis in the context of acoustic stress allowed us to establish the effects at the proteome level and identify the proteins that are particularly sensitive to this type of trauma.


Subject(s)
Decapodiformes/metabolism , Endolymph/metabolism , Proteome , Proteomics , Animals , Decapodiformes/anatomy & histology , Decapodiformes/ultrastructure , Electrophoresis, Gel, Two-Dimensional , Environmental Exposure/adverse effects , Proteomics/methods , Sound/adverse effects
17.
Actas dermo-sifiliogr. (Ed. impr.) ; 110(5): 385-392, jun. 2019. tab, graf
Article in Spanish | IBECS | ID: ibc-180927

ABSTRACT

Antecedentes y objetivo: La psoriasis es una enfermedad inmunoinflamatoria crónica de la piel muy frecuente en el mundo occidental. Muchos autores han intentado calcular su prevalencia en diversas regiones, aunque en la mayoría de los casos esta se ha obtenido mediante encuestas y existen escasas publicaciones procedentes del área mediterránea. El objetivo de nuestro estudio era analizar la prevalencia y severidad de la psoriasis en Lleida (región del noreste de España), identificar diferencias en edad y sexo, y comparar nuestros resultados con otras series europeas. Materiales y métodos: Se obtuvo una base de datos conjunta entre medicina primaria y el departamento de dermatología de toda la provincia de Lleida con datos epidemiológicos, diagnóstico de psoriasis y codificación de tratamiento. Resultados: La base de datos final comprendía a 398.701 individuos y 6.868 de ellos (1,72%) fueron codificados con el diagnóstico de psoriasis. La prevalencia de psoriasis fue significativamente mayor en hombres que en mujeres (1,88 vs. 1,56%; OR = 1,21; IC 95%: 1,15-1,27). La prevalencia más alta de psoriasis se encontró en el grupo de edad de los 61-70 años (2,90%) y la prevalencia de psoriasis en menores de 18 años fue del 0,30%. En nuestra población, el 7,27% de los pacientes fueron clasificados como psoriasis moderada-severa (499/6.868). Conclusiones: Este estudio reporta la prevalencia y severidad de la psoriasis en una muestra amplia de una región mediterránea, obteniendo la información mediante una base de datos electrónica. Además, se evidencia una prevalencia menor de psoriasis comparada con otros países europeos y una proporción de psoriasis severa (basado en criterios de tratamiento) menor que en otros estudios. Estas diferencias podrían deberse a factores genéticos, estilo de vida y dieta


Background and objectives: Psoriasis is a chronic immunoinflammatory skin disease very frequent in the western world. Several authors have tried to calculate its prevalence in different regions, although most of them obtained the data from surveys and there are few publications from Mediterranean areas. The objective of our study was to analyze the prevalence and severity of psoriasis in Lleida (a northeastern region in Spain), identify age and sex specific differences and compare our results with other European series. Materials and methods: A joint database of primary care medicine and the dermatology department was obtained from the entire province of Lleida with epidemiological data and psoriasis diagnosis and treatment codification. Results: A corrected database was obtained with 398,701 individuals and 6,868 of them (1.72%) were coded with the diagnosis of psoriasis. The prevalence was significantly higher in men than in women (1.88% vs 1.56%, OR = 1.21, 95% CI: 1.15-1.27). The highest prevalence of psoriasis was found in the 61-70 years group (2.90%) and psoriasis in population under 18 years of age was 0.30%. In our sample, 7.27% of the patients were classified as moderate-severe psoriasis (499/6,868). Conclusion: This study reports the prevalence and severity of psoriasis in a large Mediterranean region sample, obtaining the information through a electronic database. This study reveals a lower prevalence of psoriasis compared to other European countries, and the proportion of severe psoriasis (based on treatment criteria) is lower than in other studies. We emphasize that these differences could probably due to genetic background, life style and diet


Subject(s)
Humans , Male , Female , Adult , Psoriasis/epidemiology , Mediterranean Region/epidemiology , Observational Study , Cross-Sectional Studies , Spain , Electronic Health Records , Prevalence
18.
Gen Comp Endocrinol ; 275: 44-50, 2019 05 01.
Article in English | MEDLINE | ID: mdl-30716305

ABSTRACT

A significant body of literature suggests that aquatic pollutants can interfere with the physiological function of the fish hypothalamic-pituitary-interrenal (HPI) axis, and eventually impair the ability to cope with subsequent stressors. For this reason, development of accurate techniques to assess fish stress responses have become of growing interest. Fish scales have been recently recognized as a biomaterial that accumulates cortisol, hence it can be potentially used to assess chronic stress in laboratory conditions. We, therefore, aimed to evaluate the applicability of this novel method for cortisol assessment in fish within their natural environment. Catalan chub (Squalius laietanus) were sampled from two sites; a highly polluted and a less polluted (reference) site, in order to examine if habitat quality could potentially influence the cortisol deposition in scales. We also evaluated the seasonal variation in scale cortisol levels by sampling fish at three different time points during spring-summer 2014. In each sampling, blood was collected to complement the information provided by the scales. Our results demonstrated that blood and scale cortisol levels from individuals inhabiting the reference site were significantly correlated, therefore increasing the applicability of the method as a sensitive-individual measure of fish HPI axis activity, at least in non-polluted habitats. Since different environmental conditions could potentially alter the usefulness of the technique, results highlight that further validation is required to better interpret hormone fluctuations in fish scales. Scale cortisol concentrations were unaffected by habitat quality although fish from the polluted environment presented lower circulating cortisol levels. We detected a seasonal increase in scale cortisol values concurring with an energetically costly period for the species, supporting the idea that the analysis of cortisol in scales reveals changes in the HPI axis activity. Taken together, the present study suggests that cortisol levels in scales are more likely to be influenced by mid-term, intense energetically demanding periods rather than by long-term stressors. Measurement of cortisol in fish scales can open the possibility to study novel spatio-temporal contexts of interest, yet further research is required to better understand its biological relevance.


Subject(s)
Animal Scales/chemistry , Cyprinidae , Ecosystem , Hydrocortisone/analysis , Seasons , Water Quality , Animals , Cyprinidae/metabolism , Environmental Biomarkers , Environmental Pollution/analysis , Female , Fresh Water , Immunoenzyme Techniques , Male , Stress, Physiological/physiology , Water Pollution, Chemical/analysis
19.
Equine Vet J ; 51(5): 606-611, 2019 Sep.
Article in English | MEDLINE | ID: mdl-30624804

ABSTRACT

BACKGROUND: Vitiligo and melanoma are relatively common disorders in grey Pura Raza Español horses and other horse breeds with grey-coloured coats. OBJECTIVES: To determine the breed prevalence, environmental risks factors and estimate the genetic parameters for vitiligo and melanoma in Pura Raza Español horses. STUDY DESIGN: Retrospective cohort study. METHODS: We analysed data from a large worldwide population of Pura Raza Español horses. The database included the vitiligo and melanoma scores, on either a four- or six-point linear scale, of 11,436 horses. Genetic parameters were estimated using a Bayesian genetic animal model including the four associated environmental risk factors as systematic effects. Inbreeding was used as a covariate, and animal and residual effects were included as random effects. RESULTS: Of the horses included in the study, 2.8 and 20.5% showed some traces of vitiligo around the eyes and mouth, respectively, while 1.6% showed varying degrees of melanoma. Age, coat colour and inbreeding were significantly associated with the three outcomes studied. The estimated heritability for the whole population was 0.09 (s.d. +0.019), 0.44 (s.d. +0.031) and 0.13 (s.d. +0.037), for eye vitiligo score, nostril vitiligo score and melanoma scores respectively. The genetic correlations ranged from 0.42 (s.d. +0.084) between eye and nostril vitiligo score to 0.15 (s.d. +0.096) between nostril vitiligo and melanoma. MAIN LIMITATIONS: Vitiligo scores for the perianal regions were not collected. The veterinarian responsible for each assessment was not recorded. CONCLUSIONS: Vitiligo and melanoma are prevalent in this population and those environmental risk factors and genetics both have an effect on the clinical expression of the diseases. These findings may help to reduce prevalence through breeding programmes.


Subject(s)
Genetic Predisposition to Disease , Horse Diseases/genetics , Melanoma/veterinary , Vitiligo/veterinary , Aging , Animals , Female , Horse Diseases/etiology , Horses , Inbreeding , Male , Melanoma/etiology , Melanoma/genetics , Risk Factors , Skin Pigmentation , Vitiligo/etiology , Vitiligo/genetics
20.
Nat Prod Res ; 33(8): 1085-1091, 2019 Apr.
Article in English | MEDLINE | ID: mdl-29658316

ABSTRACT

It is well established that different factors affect milk composition in cows and that milk composition, in turn, affect both technological and nutritional qualities. In this respect the comprehension of the metabolic variability of milk composition in relation to the lactation time as well as to the genetic background may be of paramount importance for the agri-food industries. In the present study we investigated the variations of the metabolic profiles during lactation in milks obtained from Friesian and autochthonous races from Northern Italy by 1H NMR metabolomics. Furthermore, the external factors influencing the milk composition were minimized: the cows were breeded in the same farm, were fed with the same diet and were paired for the lactation interval and lactation stage. Our results showed a difference in milk composition between races and in relation to late lactation. The PLS-DA analysis permitted to distinguish the Friesian and autochthonous cow milks at the investigated different lactation times. Interestingly, the metabolites significantly involved into the discrimination between races appeared to be also technological property parameters, highlighting the importance of maintaining the biodiversity of cow breeds. Therefore, NMR-based metabolomics of milk could represent an informative tool to identify metabolites involved in milk quality both from a nutritional and industrial perspective.


Subject(s)
Lactation/metabolism , Magnetic Resonance Spectroscopy/methods , Metabolomics/methods , Milk/chemistry , Milk/metabolism , Animal Feed , Animals , Cattle , Female , Food Analysis/methods , Food Analysis/statistics & numerical data , Italy , Magnetic Resonance Spectroscopy/statistics & numerical data , Multivariate Analysis
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