Genetic investigation of the ubiquitin-protein ligase E3A gene as putative target in Angelman syndrome

BACKGROUND Angelman syndrome(AS)is caused by maternal chromosomal deletions,imprinting defects,paternal uniparental disomy involving chromosome 15 and the ubiquitin-protein ligase UBE3A gene mutations.However the genetic basis remains unclear for several patients.AIM To investigate the involvement of UBE3A gene in AS and identifying new potential genes using exome sequencing.METHODS We established a cohort study in 50 patients referred to Farhat Hached University Hospital between 2006 and 2021,with a strong suspicion of AS and absence of chromosomal aberrations.The UBE3A gene was screened for mutation detection.Two unrelated patients issued from consanguineous families were subjected to exome analysis.RESULTS We describe seven UBE3A variants among them 3 none previously described including intronic variants c.2220+14T>C(intron14),c.2507+43T>A(Exon15)and insertion in Exon7:c.30-47_30-46.The exome sequencing revealed 22 potential genes that could be involved in AS-like syndromes that should be investigated further.CONCLUSION Screening for UBE3A mutations in AS patients has been proven to be useful to confirm the diagnosis.Our exome findings could rise to new potential alternative target genes for genetic counseling.

Profiles of the Headspace Volatile Organic and Essential Oil Compounds from the Tunisian Cardaria draba (L.) Desv. and Its Leaf and Stem Epidermal Micromorphology

In this work, we investigated aroma volatiles emanated by dry roots, stems, leaves, flowers, and fruits of Cardariadraba (L.) Desv. growing wild in Tunisia and its aerial part essential oils (EOs) composition. A total of 37 volatileorganic compounds (96.7%–98.9%) were identified;4 esters, 4 alcohols, 7 hydrocarbons, 12 aldehydes, 5 ketones,1 lactone, 1 organosulfur compound, 2 organonitrogen compounds, and 1 acid. The hydrocarbons form the maingroup, representing 49.5%–84.6% of the total detected volatiles. The main constituent was 2,2,4,6,6-pentamethylheptane(44.5%–76.2%) reaching the highest relative percentages. Forty-two compounds were determined in thetwo fractions of EOs, representing 98.8% and 97.2% of the total oil composition, respectively. The principal componentswere hexadecanoic acid (34.6%), 6-methyl-5-hepten-2-one (18.3%), decanal (15.0%), 6,10,14-trimethyl-2-pentadecanone (13.2%), and n-pentacosane (13%). Micromorphological details of the leaf and stem epidermisusing light microscopy revealed polygonal cells with sinuous walls in the adaxial and abaxial leaf surfaces andnearly rectangular and long ones with linear and thick walls for the stem epidermis. The stomata complexes wereanisocytic in the leaf epidermis and mainly anisocytic and rarely paracytic in the stem epidermis. Non-glandulartrichomes were unbranched and long with an acute apex or short with a convex apex. The glandular ones wereidentified for the first time in this species. They were short-stalked with a large secretory head. The highest stomatalindex (17.02%) was recorded in the abaxial leaf surface. The identification of headspace volatiles and essentialoil compounds can be used to characterize this species, and the various epidermis micromorphologicalfeatures are very useful for biosystematics taxonomic studies within Brassicaceae.

Relevance of proteolysis and proteasome activation in fatty liver graft preservation: An Institut Georges Lopez-1 vs University of Wisconsin appraisal

To compare liver proteolysis and proteasome activation in steatotic liver grafts conserved in University of Wisconsin (UW) and Institut Georges Lopez-1 (IGL-1) solutions.METHODSFatty liver grafts from male obese Zücker rats were conserved in UW and IGL-1 solutions for 24 h at 4 °Cand subjected to “ex vivo” normo-thermic perfusion (2 h; 37 °C). Liver proteolysis in tissue specimens and perfusate was measured by reverse-phase high performance liquid chromatography. Total free amino acid release was correlated with the activation of the ubiquitin proteasome system (UPS: measured as chymotryptic-like activity and 20S and 19S proteasome), the prevention of liver injury (transaminases), mitochondrial injury (confocal microscopy) and inflammation markers (TNF 1 alpha, high mobility group box-1 (HGMB-1) and PPAR gamma), and liver apoptosis (TUNEL assay, cytochrome c and caspase 3).RESULTSProfiles of free AA (alanine, proline, leucine, isoleucine, methionine, lysine, ornithine, and threonine, among others) were similar for tissue and reperfusion effluent. In all cases, the IGL-1 solution showed a significantly higher prevention of proteolysis than UW (P < 0.05) after cold ischemia reperfusion. Livers conserved in IGL-1 presented more effective prevention of ATP-breakdown and more inhibition of UPS activity (measured as chymotryptic-like activity). In addition, the prevention of liver proteolysis and UPS activation correlated with the prevention of liver injury (AST/ALT) and mitochondrial damage (revealed by confocal microscopy findings) as well as with the prevention of inflammatory markers (TNF1alpha and HMGB) after reperfusion. In addition, the liver grafts preserved in IGL-1 showed a significant decrease in liver apoptosis, as shown by TUNEL assay and the reduction of cytochrome c, caspase 3 and P62 levels.CONCLUSIONOur comparison of these two preservation solutions suggests that IGL-1 helps to prevent ATP breakdown more effectively than UW and subsequently achieves a higher UPS inhibition and reduced liver proteolysis.

Patterns of genetic and morphometric diversity in the marbled crab(Pachygrapsus marmoratus, Brachyura, Grapsidae)populations across the Tunisian coast

The present study reports on population structure analysis of the marbled crab Pachygrapsus marmoratus （Fabricius, 1787） from the Tunisian coast, an appropriate location to study biogeographical processes because of the presence of a well-known discontinuous biogeographic area （the Siculo-Tunisian Strait）. Patterns of morphological and genetic variation of this highly dispersive and continuously distributed decapod species were assessed among its geographically close populations which cover almost the entire Tunisian coastline. A total of 386 specimens from nine sites were collected and examined for morphometric variability at 14 morphometric traits. The results of multivariate analyses of linear morphometric traits showed the existence of sexual dimorphism in this species by PERMANOVA （Permutational multivariate analysis of variance）. In addition, both CDA （Canonical discriminant analysis） and NPMANOVA （Non parametric MANOVA test） analyses revealed statistically significant differences among the studied locations for both sexes. Overall, the outcome of CDA analysis showed that over 87% of individuals could be assigned correctly to three regional groups in both sexes （North, Center and South）. Specifically, SIMPER （Similarity Percentages） analysis showed that carapace length, carapace width and merus length were major contributors to the morphometric separation between populations. The pattern of phenotypic variation suggested by morphometric analyses was found to be highly discordant with that suggested by the analysis of a mitochondrial marker （cytochrome oxidase I, COI）. Indeed, the results inferred from restriction fragment analysis of the COI in 180 crabs, suggested high genetic homogeneity. Very low levels of haplotype diversity （h） were found in almost all the studied populations, associated with non significant genetic distances for nearly all population comparisons. Explanations to these morphometric and mtDNA patterns as well as the discrepancy between them are discussed.

Aqueous and organic extracts of Trigonella foenum-graecum L.inhibit the mycelia growth of fungi

Aqueous extracts from various plant parts of fenugreek(3%)(aerial parts:leaves and stems(LS),roots(R),ground seeds(GS)and not ground seeds(NGS))and petroleum ether,ethyl acetate and methanolic fractions of the aerial parts were assayed to determine their antifungal potential against Botrytis cinerea,Fusarium graminearum,Alternaria sp.,Pythium aphanidermatum,and Rhizoctinia solani.All fenugreek plant parts showed antifungal potential and the magnitude of their inhibitory effects was species and plant parts d...

Impact of the Liaison Committee of Food and Nutrition on the Quality of Patients＇ Meals

At hospital, nutrition represents an important value of care, particularly for patients at risk. However, it is observed that for various reasons, the restoration of the hospitalized patients is often neglected in the privileges of the medical care. The establishment of a Liaison Committee of Food and Nutrition （LCFN） within a health establishment has shown according to several works, its positive impact in improving the hygienic quality and nutritional dishes served to patients. In this framework, we conducted a quasi-experimental study into CHU F hached Sousse of Tunisia type （before/after, creation LCFN） in order to assess the role of such structure （LCFN） in the qualification and the improvement of patients＇ food. Our study was based during the two phases （2007/2010） on the same methodology. Thus we have conducted an audit of observation of hygiene practices along the distribution chain of patient＇s meals and the samples for microbiological analyzes from food, surfaces, equipment and personnel＇s hands.The results obtained have shown, in one hand, a degradation of the average rate of contamination for the bacteriological analyzes, and in the other hand, an evolution of the rate of hygiene standards respected. According to this study, the role played by the LCFN is becoming more and clearer in the improvement of the hygienic quality of patients＇ dishes without forgetting the impact of improving the nutritional quality and Hedonics.

Lack of mitochondrial genetic structure in the endangered giant clam populations of Tridacna maxima(Bivalvia: Cardiidae:Tridacninae) across the Saudi Arabian coast

The present investigation focuses on population genetic structure analysis of the endangered giant clam species Tridacna maxima across part of the Red Sea,with the main aim of assessing the influence of postulated potential barriers to gene flow(i.e.,particular oceanographic features and marked environmental heterogeneity)on genetic connectivity among populations of this poorly dispersive bivalve species.For this purpose,a total of 44 specimens of T.maxima were collected from five sampling locations along the Saudi Arabian coast and examined for genetic variability at the considerably variable mitochondrial gene cytochrome c oxidase I(COI).Our results revealed lack of population subdivision and phylogeographic structure across the surveyed geographic spectrum,suggesting that neither the short pelagic larval dispersal nor the various postulated barriers to gene flow in the Red Sea can trigger the onset of marked genetic differentiation in T.maxima.Furthermore,the discerned shallow COI haplotype genealogy(exhibiting high haplotype diversity and low nucleotide diversity),associated with recent demographic and spatial expansion events,can be considered as residual effect of a recent evolutionary history of the species in the Red Sea.

Antimicrobial activity of Tunisian Euphorbia paralias L.

Objective: To examine the potential antimicrobial activity of Euphorbia paralias L. (Euphorbiaeae) leaves and stems extracts. Methods: The antimicrobial activity was tested against six microbial strains:Escherichia coli ATCC 8739, Bacillus subtilis ATCC 6633, Salmonella enterica CIP 8039, Staphy-lococcus aureus ATCC 6538, Pseudomonas aeruginosa ATCC 9027 and Candida albicans ATCC 90028 by two different methods, the disk method and the dilution method. Results: Our results showed the important antimicrobial activity of the chloroform extract of the stems towards the majority of the strains by using both methods. Bacillus subtilis was the most sensitive strain (MIC=MBC=15μg/mL). Conclusion: Thus, some extracts of Euphorbia paralias can be used in the treatment of infectious diseases caused by microbes.

Evaluation of Risk Management of Foodborne Infection in Hospital

As being at the head of the establishment, the hospital manager＇s major concern is food safety in his hospital. Unfortunately, because of their health weaknesses, patients are more susceptible to foodborne illness than any other population group As match as food prepared according to less stringent hygiene＇s rules can infect or intoxicate more than it would be in a healthy population, as hygiene＇s rules must be observed in a particularly strict way, at the kitchen where meals are prepared for health care facilities. In fact, hospitals＇ food hygiene＇s rules are those who defined catering. They are made to avoid food contamination and microbial growth throughout the food chain from raw materials＇ delivery to the consumer. The main objective of taking the meal as a patients＇ tracer is to detect and study the infection＇s risk associated with feeding at hospital, and to provide practical tools to better manage this risk. Methodology was based on an analysis of the last six years＇ work concerning hygiene＇s audits and bacteriological analyses. The results showed that the food infectious risk has an important frequency, on one side; on another, they were used as a standard model of such risk＇s management.

Experimental Design Approach with Response Surface Methodology for Removal of Indigo Dye by Electrocoagulation

Treatment of Indigo dye (leuco form), reduced in the industrial conditions of the SITEX (Textile Industrial Company), by a batch electrocoagulation using aluminum electrodes. Response Surface Methodology (RSM) and Box-Behnken design were used to optimize for Color Removal (CR(%)). Our results showed that the quadratic second order equation provided the best correlation for the decolorization of Indigo dye (CR(%)). On the other hand, the ANOVA analysis proved the large interaction between the current intensity and the initial concentration of the dye. Experiments were conducted to find the desired conditions for removal of particular concentration of the dye and lower Operation Cost. The results showed that CR(%) = 88.3% (R2) of color removal for initial dye concentration of 12.31 mg/l, with a current density of 2.81 A/m2, solution concentration of NaCl of 2.67 g/l. Under these conditions, Electrical Energy Consumption (EEC) and Electrode Consumption (EMC) and Operation Cost were 0.01999 kWh/m3 (R2 = 93.1%), 0.00142768 Kg/m3 (R2 = 79.4%) and 0.000558 US$/m3, respectively.

Pollen morphology of some Tunisian species from genus of Trigonella L.

The pollen morphology of lbur species from the genus Trigonella L. （Fabaceae） distributed in Tunisia （T. foenum-graecum, T. maritima, T. stellata and T. monspeliaca） was studied with light and scanning electron microscope （SEM）. Results showed that the pollen morphology of these four species was not homogenous in relation to shape, dimensions and ornamentation. The pollen grains were 3-zonocolporate, radially symmetrical, isopolar, prolate to perprolate type （P/E = 1.45-2.09）. Medium in size, elliptical or rectangular-obtuse-convex in equatorial view and circular in polar view. Ectoapertures-colpi： long, medium width, with acute ends, endoapertures-pork large, circular to lolongate, protruding in mesocolpium. The rectum was reticulate to perforate reticulate with ± irregular pattern of muri, except for T. monspeliaca which ornamentation was relatively homogeneous with a loose network and psilate area. Ornamentation with lumina different in size was most clearly expressed in mesocolpium while the apocolpium and the area just along the apertures was psilate or with small scattered puncta. The exine thickness was 1.10-2.42 μtm.

Genetic and morphological divergence in the purple sea urchin Paracentrotus lividus(Echinodermata,Echinoidea)across the African Mediterranean coast

The present investigation focuses on population structure analysis of the purple sea urchin Paracentrotus lividus across the African Mediterranean coast, with the main aim of assessing the influence of the Siculo-Tunisian Strait on gene flow disruption in this highly dispersive echinoid species. For this purpose, patterns of morphological and genetic variation were assessed among its populations from the western and eastern Mediterranean coasts. A total of 302 specimens from seven Tunisian sites were collected and examined for morphometric variability at twelve morphometric traits. Concordant results, inferred from CDA（canonical discriminant analyses）, pairwise NPMANOVA（non parametric multivariate analysis of variance） comparisons and MDS（multidimensional scaling） plot, unveiled significant inter-population differences in the measured traits among the studied populations. Furthermore, the combined use of the one way ANOSIM（analysis of similarities） and the Discriminant/Hotelling analysis allowed unravelling two morphologically differentiated groups assigned to both western and eastern Mediterranean basins. The SIMPER（similarity percentages） routine analysis showed that total dry weight, test diameter and spine length were major contributors to the morphometric separation between locations and between groups. Pattern of phenotypic divergence discerned in P. lividus across the Siculo-Tunisian Strait is interestingly in congruence with that inferred from the genetic investigation of the purple sea urchin populations from the same region based on the analysis of the mt DNA COI（cytochrome oxidase I） gene in 314 specimens from nineteen locations covering a wider geographic transect, streching westward to the Algerian coast and eastward to the Libyan littoral. The specific haplotypic composition characterizing each Mediterranean basin,as inferred from the minimum spanning network, confirmed the geographic partioning of genetic variation, as revealed by F-statistics and AMOVA（analysis of molecular variance） analyses, yielding significant genetic differentiation between eastern and western Mediterranean populations. The newly detected phylogeographic patterns, observed for the first time in P. lividus throughout the explored distribution range, suggest the involvement of different biotic and abiotic processes in shaping such variation, and provide evidence that a large and geographically exhaustive dataset is necessary to unveil phylogeographic structure within widespread marine species, previously cathegorized as panmictic in part of their distribution range.

Antioxidant activity, volatile and nonvolatile composition of twoPhoenix dactylifera L. by-products

Objective:To examine the nonvolatile components of the date pulp and seeds of two minor Tunisian date palm cultivars Arechti and Korkobbi at besser stage and to investigate their antioxidant activity, their phytochemical compounds and their aromatic volatiles composition. Methods:The physico-chemical properties, the aroma composition, the mineral profile, the phytochemical content and thein vitro antioxidant activity of those two common date palm fruit varieties and those of their seeds were evaluated. All measurements were made at the besser stage, which showed the highest amount of secondary metabolites. Results:New volatiles compounds are detected in both flesh and seeds and the results revealed that those by products are a suitable source of mineral content and have a high potential of natural antioxidants and good antiradical capacities. Conclusions: Thanks to their characteristics, they could potentially be considered as a cheap resource for new functional food ingredient and novel pharmaceutical applications.

Isolation of bioactive antioxidant compounds from the aerial parts of Allium roseum var.grandiflorum subvar.typicum Regel.

Objective:To assess the antioxidant activity of aerial parts of Allium roseum var.grandiflorum subvar.typicum Regel.(A.roseum var.grandiflorum subvar.typicum Regel.)for the first time,as well as to isolate the main bioactive compounds.Methods:The chloroformic extract of Allium roseum(A.roseum)and their fractions obtained by subjection to a chromatographic study were tested for their antioxidant activities by using 2,2-diphenyl-2-picrylhydrazyl and 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid)assays.An activity-guided purification was conducted to isolate five compounds in pure form where their structures were identified by means of nuclear magnetic resonance analyses,including 1D and 2D nuclear magnetic resonance experiments.Results:The evaluation of the antioxidant activity of chloroformic extract and their fractions from A.roseum var.grandiflorum subvar.typicum Regel.showed interesting results.The active chloroformic extract afforded five isolated compounds where their structures were identified as β-sitosterol(1),chrysoeriol(2),luteolin(3),apigenin(4),andβ-sitosterol 3-O-β-D-glucoside(5).All the compounds were isolated for the first time from the A.roseum var.grandiflorum subvar.typicum Regel.The three flavonoids(2–4)exhibited the highest antioxidant activity with IC_(50) values of 62.28,21.26 and 513.42μg/mL,respectively(2,2-diphenyl-2-picrylhydrazyl assay)and 218.00,73.50 and 877.66μg/mL,respectively[2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid)assay].An important value of trolox equivalent antioxidant capacity(2.10 mmol/L)was reported for luteolin(3).Conclusions:These results may suggest that the A.roseum var.grandiflorum subvar.typicum Regel.have great potential as a source of a natural preservative ingredient in beneficial for natural health products.

Total phenolic compounds,antioxidant potential and α-glucosidase inhibition by Tunisian Euphorbia paralias L.

Objective:To examine the potential antioxidant and anti-α-glucosidase inhibitory activities of Tunisian Euphorbia paralias L.leaves and stems extracts and their composition of total polyphenol and flavonoids.Methods:The different samples were tested for their antiradical activities by using 2,2’-azinobis(3-ethylbenzothiazoline-6-sulfonic acid)(ABTS)and 1,1-diphenyl-2-picrylhydrazyl(DPPH)assays.Inα-glucosidase activity,α-glucosidase(0.3 IU/mL)and substrate,2500μmol/L p-nitrophenylα-D-glucopyranoside were used;absorbance was registered at 405 nm.Results:The leaves acetonic extract exhibited the strongestα-glucosidase inhibition[IC_(50)=(0.0035±0.001)μg/mL],which was 20-fold more active than the standard product(acarbose)[IC_(50)=(0.07±0.01)μg/mL].Acetonic extract of the leaves exhibited the highest quantity of total phenolic[(95.54±0.04)μg gallic acid equivalent/mg]and flavonoid[(55.16±0.25)μg quercetin equivalent/mg].The obtained findings presented also that this extract was detected with best antioxidant capacity[IC_(50)=(0.015±0.01)μg/mL]against DPPH and a value of IC_(50)equal to(0.02±0.01)μg/mL against ABTS.Positive relationship between polyphenolic content of the tested Euphorbia paralias L.leaves and stems extracts and its antioxidant activity(DPPH and ABTS)was detected.Elevated positive linear correlation was got between ABTS and total phenolic(R^(2)=0.751).Conclusions:The findings clearly demonstrate that the use of a polar solvent enables extraction of significant quantities of phenol compounds and flavonoids.