Francesco Palmas

Perinatal Asphyxia: A Review from a Metabolomics Perspective

Perinatal asphyxia is defined as an oxygen deprivation that occurs around the time of birth, and may be caused by several perinatal events. This medical condition affects some four million neonates worldwide per year, causing the death of one million subjects. In most cases, infants successfully recover from hypoxia episodes; however, some patients may develop HIE, leading to permanent neurological conditions or impairment of different organs and systems. Given its multifactor dependency, the timing, severity and outcome of this disease, mainly assessed through Sarnat staging, are of difficult evaluation. Moreover, although the latest newborn resuscitation guideline suggests the use of a 21% oxygen concentration or room air, such an approach is still under debate. Therefore, the pathological mechanism is still not clear and a golden standard treatment has yet to be defined. In this context, metabolomics, a new discipline that has described important perinatal issues over the last years, proved to be a useful tool for the monitoring, the assessment, and the identification of potential biomarkers associated with asphyxia events. This review covers metabolomics research on perinatal asphyxia condition, examining in detail the studies reported both on animal and human models.

Genetic characterization and artificial reproduction attempt of endemic Sardinian trout Salmo trutta L., 1758 (Osteichthyes, Salmonidae): Experiences in captivity

Abstract Native species of Sardinian brown trout (Salmo trutta) have declined drastically because of the introduction of non-native salmonids, overharvesting, and habitat degradation. One approach to conservation of declining species is to establish new populations through repopulation with genetically certified fish. In this study, samples of brown trout from seven rivers in Sardinia were analysed to look for the presence of pure individuals of the endemic Sardinian trout. Analyses of the mitochondrial DNA control region and the LDH-C1* gene revealed the presence of non-endemic and hybrid individuals in four of the seven rivers analysed. In addition, we reared a stock of native Sardinian trout at the experimental fish farm of Sadali and had them reproduce artificially. This experiment provided more knowledge about the incubation phase, hatching, reabsorption of the vitelline sac and the initial phases of growth, which can be used to develop more optimal management of this resource.

The choice of amniotic fluid in metabolomics for the monitoring of fetus health

ABSTRACT Amniotic fluid (AF) is a biological fluid in which metabolite transport is regulated by the placenta, the permeable skin, fetal lung egress and gastric fluid. During pregnancy, the composition of AF changes from similar to the interstitial fluid of the mother, to a more complex system, influenced by the fetus’s urine. Since AF reflects the mother’s and the fetus’s health status at the same time, it may be an important diagnostic tool for a wider spectrum of clinical conditions. Indeed, the metabolic characterization of AF in relation to pathological occurrences may lead to the discovery of new biomarkers for a better clinical practice. For this reason, metabolomics may be the most suitable strategy for this task. In this review, research works on metabolomic AF analysis are discussed according to the morbidity of interest, being preterm birth/labor, gestational age and diabetes and fetal malformations, along with a number of other important studies.

Antennular Morphology and Contribution of Aesthetascs in the Detection of Food-related Compounds in the Shrimp Palaemon adspersus Rathke, 1837 (Decapoda: Palaemonidae)

Shrimp are an essential ecological component of marine ecosystems, and have commercial importance for human consumption and aquaculture. Like other decapod crustaceans, shrimp rely on chemical senses to detect and localize food resources by means of chemosensilla that are located mainly on the cephalothoracic appendages. Using the shrimp Palaemon adspersus, a model organism with omnivorous feeding behavior, we aimed to provide comparative information on the role of aesthetascs, antennular sensilla, and flicking behavior in food detection. To this end, we examined i) the morphology of antennular sensilla by field emission scanning electron microscopy, ii) the shrimp’s sensitivity to a number of food-related compounds (amino acids and sugars) by means of whole-animal bioassays, and iii) the contribution of the aesthetasc sensilla to food detection. Our results showed that, aside from the aesthetascs, only three other main morphotypes of setae with chemoreceptive features were present in the antennules, thus accounting for relatively simple sensillar equipment. Nevertheless, we found broad-spectrum sensitivity of the shrimp to a number of amino acids (i.e., isoleucine, leucine, methionine, phenylalanine, glycine, tryptophan, cysteine, and tyrosine) and carbohydrates (trehalose, maltose, cellobiose, and fructose) that was consistent with the omnivorous or scavenging habits of the animal. Although aesthetasc ablation attenuated flicking behavior in a chemical stimulus-independent manner, success in detection and short-range localization of food did not rely on the presence of aesthetasc sensilla. This finding confirms the existence of a non-aesthetasc alternative pathway for feeding, with functional redundancy in simple generalist feeder models such as shrimp.

Demersal assemblages in two trawl fishing lanes located on the Baronie Seamount (Central Western Mediterranean)

This work focuses on the species associations of the Baronie Seamount (north-eastern Sardinia, Italy), according to variations in depth and time of day. The aim was to highlight the potential vertical movements of the species and to compare these results to existing data from a submarine canyon. Twenty-two samples were taken at different depths and times along two trawl lines over the seamount. A total of 94 species were caught; among these, 4 8 bony fish, 9 cartilaginous fish, 13 molluscs and 24 crustaceans were selected for analysis. Cluster analysis showed 3 groups in which both depth and time of day play an important role in grouping. Nine species showed some daily and nocturnal movement, probably linked to trophic requirements. The daily movements according to size and sex of the shrimps Aristaeomorpha foliacea and Aristeus antennatus were also studied. For these two populations, the analysis showed an uneven distribution and the diel cycle appears to involve only the females. These species seem to adapt their life cycle to the geomorphology of the sea bottom. The diurnal movements from the base of the seamount to the edge of the continental shelf increase the range of the species distribution. In fact, the seamount, due to its geomorphological conformation, offers these species the opportunity to very quickly span a considerable range of depths. These movements are known to occur in the continental shelf and slope, but in the seamount they are broadened and can be studied more easily.

Urinary metabolomic analysis to identify preterm neonates exposed to histological chorioamnionitis: A pilot study

Objective Chorioamnionitis is a leading cause of preterm birth worldwide, with higher incidence at lower gestational ages. An early and reliable diagnosis of histological chorioamnionitis (HCA) in preterm infants may be helpful in guiding postnatal management, especially the administration of prophylactic antibiotics to prevent early-onset sepsis. The main aim of this study was to investigate metabolomic analysis of urines collected in the first 24 hours of life as diagnostic tool of HCA. Methods Gestational age-, birth weight-, delivery mode- and sex- matched (1:2) preterm neonates (< 35 weeks’ gestation) born to mothers with or without HCA were enrolled from an observational study. Gas chromatography-mass spectrometry (GC-MS)-based metabolomic analysis was performed on urine samples non-invasively collected in the first 24 hours of life. Univariate analysis, partial least square discriminant analysis (PLS-DA) and its associated variable importance in projection (VIP) score were performed. The most affected metabolic pathways were examined by Metabolite Sets Enrichment Analysis (MSEA). Results Fifteen cases (mean GA 30.2 ± 3.8 weeks, mean BW 1415 ± 471.9 grams) and 30 controls (mean GA 30.2 ± 2.9 weeks, mean BW 1426 ± 569.8 grams) were enrolled. Following univariate analysis, 29 metabolites had a significantly different concentration between cases and controls. The supervised PLS-DA model confirmed a separation between the two groups. Only gluconic acid, an oxidation product of glucose, was higher in cases than in controls. All other VIP metabolites were more abundant in the control group. Glutamate metabolism, mitochondrial electron transport chain, citric acid cycle, galactose metabolism, and fructose and mannose degradation metabolism were the most significantly altered pathways (P < 0.01). Conclusions For the first time, urinary metabolomics was able to discriminate neonates born to mothers with and without HCA. The identification of specifically altered metabolic pathways may be helpful in understanding metabolic derangement following chorioamnionitis.

Rare disease: a focus on metabolomics

ABSTRACT Introduction: A disease or disorder is defined rare when it affects less than 1 in 2000 subjects. 400 million people worldwide are living with a rare disease; approximately 50% of these patients are children, and 30% of them die within the first 5 years of life. The wide aetiology diversity leads to high variability in symptomatology, and clinical manifestations and consequently to challenging diagnosis. New experimental approaches are therefore recommended to improve diagnosis, and for the development of new therapeutic strategies. In the last years, metabolomics have allowed to better understand the metabolic processes associated with several rare diseases. Areas covered: This review describes the use of metabolomics for the study of the metabolic alterations related to some of the most studied pediatric rare disorders. Metabolomics, by means of 1H NMR spectroscopy and MS spectrometry, allows the identification and the quantification of a large number of metabolites (biomarkers) in different biofluids. Expert opinion: Metabolomics proved to be a powerful tool for the clinical investigation of rare diseases. The power of discrimination seems to be reliable for all the considered biofluids (urine, blood, and amniotic fluid). Standardization of the analytical and statistical steps is still needed for further studies with larger populations and meta-analysis.

Sensing with the legs: contribution of pereiopods in the detection of food-related compounds in the red swamp crayfish Procambarus clarkii

The red swamp crayfish Procambarus clarkii (Girard, 1852) is one of the most hazardous invasive alien species of freshwater habitats. Like other decapod crustaceans, crayfish rely on chemical senses to detect and localize food resources. By way of extracellular nerve recordings coupled with behavioural bioassays, we investigated the sensitivity spectra of the walking leg chemoreceptors of P. clarkii in response to different food-related compounds. Recordings from isolated legs confirmed a marked sensitivity of the legs to trehalose, cellobiose, sucrose, maltose, glycine, and leucine. Some sensitivity to glucose, fructose, asparagine, and taurocholic acid was also found. Under confined experimental conditions, the behavioural bioassays excluded involvement of antennules in the detection of food-related compounds, thus emphasizing the role of the legs as the main short-distance, broad-spectrum sensors involved in feeding. Such information could be valuable for the identification of key chemicals aimed at the future development of strategies for crayfish population control programs.

Urine metabolome analysis by gas chromatography–mass spectrometry (GC–MS): Standardization and optimization of protocols for urea removal and short-term sample storage

BACKGROUND Before derivatization, urine analyzed by gas chromatography-mass spectrometry (GC-MS) requires the complete removal of urea to avoid interferences. We aimed at establishing the most effective sample pretreatment for urea removing; moreover, we explored the impact of two short-term sample storage conditions on urine metabolome. METHODS 92 aliquots were obtained from a single sample collected from a healthy adult; they were divided into 6 groups. Group 1 consisted of untreated aliquots while groups 2-6 differed from each other for the addition of various defined urease solution volumes combined with either 30 min or 1-hour sonication time. Urine sample storage was tested by comparing 20 fresh aliquots analyzed after collection with 20 aliquots frozen at -80 °C for 72 h. RESULTS the most effective protocol consisted of the combination between 200 μL urease solution with 1-h sonication time; urease solution volumes >200 μL increase the risk to underestimate metabolite peaks because of sample dilution. Short-term storage of samples at -80 °C pointed out significant changes in the urine metabolic profile compared with that of fresh samples. CONCLUSIONS our study confirms the importance of urea removal for a reliable recognition and quantitation of metabolites; urine short-term storage at -80 °C should be carefully reconsidered.

Multivariate data validation for investigating primary HCMV infection in pregnancy

We reported data concerning the Gas Chromatography–Mass Spectrometry (GC–MS) based metabolomic analysis of amniotic fluid (AF) samples obtained from pregnant women infected with Human Cytomegalovirus (HCMV). These data support the publication “Primary HCMV Infection in Pregnancy from Classic Data towards Metabolomics: an Exploratory analysis” (C. Fattuoni, F. Palmas, A. Noto, L. Barberini, M. Mussap, et al., 2016) [2]. GC–MS and Multivariate analysis allow to recognize the molecular phenotype of HCMV infected fetuses (transmitters) and that of HCMV non-infected fetuses (non-transmitters); moreover, GC–MS and multivariate analysis allow to distinguish and to compare the molecular phenotype of these two groups with a control group consisting of AF samples obtained in HCMV non-infected pregnant women. The obtained data discriminate controls from transmitters as well as from non-transmitters; no statistically significant difference was found between transmitters and non-transmitters.