Nisin-based therapy: a realistic and eco-friendly biocontrol strategy to contrast Xylella fastidiosa subsp. pauca infections in planta.

The lack of sustainable strategies for combating Xylella fastidiosa (Xf) highlights the pressing need for novel practical antibacterial tools. In this study, Lactococcus lactis subsp. lactis strain ATCC 11454 (L. lactis), known for its production of nisin A, was in vitro tested against Xf subsp. pauca. Preliminary investigations showed that nisin A was involved in a strong antagonistic activity exhibited by L. lactis against Xf. Thus, the efficacy of nisin A was comprehensively assessed through a combination of in vitro and in planta experiments. In vitro investigations employing viable-quantitative PCR, spot assay, turbidity reduction assay, fluorescence microscopy, and transmission electron microscopy demonstrated nisin's robust bactericidal effect on Xf at a minimal lethal concentration of 0.6 mg/mL. Moreover, results from fluorescence and transmission electron microscopies indicated that nisin directly and rapidly interacts with the membranes of Xf cells, leading to the destruction of bacterial cells in few minutes. In in planta tests, nisin also demonstrated the ability to tackle Xf infections within Nicotiana benthamiana plants that remained asymptomatic 74 days post inoculation. Furthermore, RPLC-ESI-MS/MS analyses showed that nisin translocated to all parts of the plants and remains intact for up to 9 days. For the first time, this study underscores the nisin-based strategy as a realistic and eco-friendly approach to be further investigated against Xf infections in the field.

Synecology of Lagoecia cuminoides L. in Italy and evaluation of functional compounds presence in its water or hydroalcoholic extracts.

Lagoecia cuminoides L. is a very rare and threatened taxon in Italy, never studied before for its ecology and potential use for human consumption. Furthermore, few data are available on the biological activities of its metabolites. A phytosociological study was carried out in the only two Italian sites, and its state of conservation was also evaluated according to the IUCN (International Union for Conservation of Nature) protocol. The collected plant material was used to make two types of extracts: hot water infusion to evaluate the use of this plant as tea and hydroalcoholic extraction to evaluate the use of it in herbal liqueur preparation. The presence of functional compounds in the extracts were investigated by gas and liquid chromatography coupled to mass spectrometry techniques. Ten non volatiles compounds were identified in the extracts, most of which derivatives of quercetin. Thirty-five volatiles compounds were also identified in the plant aerial part and extracts belonging to the chemical class of terpenoids, and among them ß-farnesene, thymol, γ-terpinene and p-cymene were the most abundant. The species is characterized by compounds known for their health effects and for its potential applications for human consumption, being this species already used as decoction in some countries of Middle East. Thanks to its characteristic behaviour to grow in limiting pedoclimatic conditions this species can be potentially used in organic farms situated in rural marginal areas.

A proteomic study of "Coppa Piacentina": A typical Italian dry-cured Salami.

Coppa Piacentina is considered a peculiar dry cured salami, since it is manufactured by the entire neck muscles stuffed and matured in natural casings, the same as dry cured ham and fermented dry cured sausages. In this work the proteolysis of external and internal portions was investigated by a proteomic approach and by amino acids analysis. Samples of "Coppa Piacentina" were analyzed at 0 days and after 5 and 8 months of ripening through mono- and two-dimensional gel electrophoresis. Image analysis of 2D electrophoretic maps indicated a more intense enzyme activity on the external part, mainly due to endogenous enzymes. They favored, respectively, myofibrillar or sarcoplasmic proteins at 5 or 8 months of ripening. Free amino acids determination proved that lysine and glutamic acid were the most represented ones, followed by a free amino acids sequence like that of dry cured ham. The peculiarities of "Coppa Piacentina" were characterized by a slow proteolysis, due to sacking and binding of the whole cut of the pork neck.

The Effect of Ultrasound Treatment in Winemaking on the Volatile Compounds of Aglianico, Nero di Troia, and Primitivo Red Wines.

An ultrasound (US) treatment was applied during the vinification of three different red grape cultivars with the aim of assessing the impact on the volatile profile of the wines. A robust solid phase micro extraction coupled with gas chromatography mass-spectrometry (SPME-GC-MS) method was developed in order to fix the best parameters for optimizing the volatile organic compound (VOC) recovery. A 15% NaCl solution was added to the samples to increase the salting-out effect, the time/temperature were appropriately selected, and the matrix effect was evaluated by comparing synthetic and real matrices. In addition, external calibration curves were used to quantify the single volatile compounds. The analyses of the wine samples at 7 and 14 months of aging revealed that US exerted the highest effect on Aglianico, which had the highest amount of total VOC. US Nero di Troia showed similar results after 14 months of aging, while Primitivo was not affected by the treatment. Nevertheless, from discriminant analysis, a clear separation was observed between the control and ultrasound-treated wines for all three cultivars, with ethyl decanoate, ethyl isopentyl succinate, and butyric acid having the highest discriminant coefficients. In conclusion, the obtained results indicated that the effect of US treatment on the VOC profile of the wine considered in the experimentation is cultivar-dependent.

Carotid endarterectomy with saphenous vein patch angioplasty: a single-center experience.

BACKGROUND: When performing a conventional CEA it is recommended the use of patch angioplasty (PA), since previous meta-analyses have shown PA to be superior to primary closure (PRC) in terms of stroke and restenosis rates. Different materials patches can be employed although none of them has been proved to be superior. Although autologous veins are potentially more resistant to immediate thrombosis as well as infection, cons may be represented by patch rupture and late dilatation. Aim of this study is to evaluate immediate and long-term results of CEA with saphenous vein patch angioplasty (SVPA) in a single-center experience. METHODS: A retrospective study was performed analyzing all patients undergoing CEA with SVPA at our institution from January 2012 to March 2020. CEA was performed in symptomatic patients with 50-99% carotid stenosis degree or asymptomatic patients with 70-99% stenosis degree. Exclusion criteria were critical limb ischemia, varicose disease, unavailability of saphenous veins, vein diameter <3.5 mm. All CEAs were performed under general anesthesia with routine shunting. Primary endpoints were perioperative stroke, death, carotid thrombosis and hematoma requiring surgery rates. Secondary endpoints included the rate of recurrent stenosis >70%, patch aneurysm/rupture/infection at follow-up. RESULTS: Overall, 488 interventions were performed on 461 patients. Most patients were male (77.8%) with a mean age of 71.2±8.3 years. Thirty-day mortality and stroke rates were 0.4% and 1.2% respectively. Carotid thrombosis occurred in five patients (1%). Five patients (1%) developed a surgical site hematoma requiring surgical drainage. At a mean follow-up of 34.4±25.8 months 12 restenoses (2.5%) were detected. Five-year freedom from restenosis rate was 96.7%. Restenosis at follow-up was more frequent in patients who had contralateral carotid stenosis (P=0.019). Two patients (0.4%) developed carotid patch aneurysmal degeneration at a mean follow-up of 78.7 months. No infection nor patch disruption were detected. CONCLUSIONS: CEA with SVPA resulted safe and effective in terms of early and late results. The perioperative complications rates we recorded were quite similar to those reported by other larger reviews and meta-analyses.

Microplastics in water surface and in the gastrointestinal tract of target marine organisms in Salento coastal seas (Italy, Southern Puglia).

Microplastic pollution is a major global environmental threat that has attracted increasing interest from the scientific community over the past decade. The semi-closed and highly urbanized Mediterranean Sea has been investigated since 2012, in several specific studies that have identified it as a target hotspot for microplastic contamination. The marine coastal zone of the Salento peninsula (Apulia, Italy) has peculiar geographical and hydrodynamic features, although there are few published data detailing the level of microplastics present in this area. The present manuscript contains both data on the concentration of microplastics in surface waters and the level of microplastics ingested by selected marine organisms in the Salento coastal zone. Microplastics floating on the water surface were monitored during Autumn 2020 and Spring 2021 using neuston Manta net at three different distances from the coasts (Lizzano, Gallipoli and Otranto). The level of microplastic ingestion was monitored in fish species (Sardina pilchardus, Boops boops, Mullus barbatus) and in mussels (Mytilus galloprovincialis). Episodic peaks of microplastic concentrations were found on the sea surface during transects performed in the 3 nautic miles from the seashore. High values of ingested microplastics were found in S. pilchardus. and B. boops (5.4 and 4.6 items/individual respectively). A higher concentration of microplastics was detected in the Adriatic Sea than in the Ionian Sea by comparing the gastrointestinal tract of S. pilchardus and B. boops, in the monitored areas. These results are correlated with the concentration of floating microplastics, although this last result is not validated by statistical analysis. These results support the effectiveness of S. pilchardus and B. boops used as targets in monitoring activity for these pollutants. Results show a worrying increase in the concentration of microplastics on the sea surface and in the gastrointestinal tract of the target species compared to data reported in the literature.

Cheese ripening in nonconventional conditions: A multiparameter study applied to Protected Geographical Indication Canestrato di Moliterno cheese.

A multiparameter study was performed to evaluate the effect of fondaco, a traditional ripening cellar without any artificial temperature and relative humidity control, on the chemical, microbiological, and sensory characteristics of Protected Geographical Indication Canestrato di Moliterno cheese. Ripening in such a nonconventional environment was associated with lower counts of lactococci, lactobacilli, and total viable bacteria, and higher presence of enterococci, in comparison with ripening in a controlled maturation room. Moreover, fondaco cheese underwent accelerated maturation, as demonstrated by faster casein degradation, greater accumulation of free AA, and higher formation of volatile organic compounds. Secondary proteolysis, as assessed by liquid chromatography-mass spectrometry of free AA and low molecular weight peptides, did not show any qualitative difference among cheeses, but fondaco samples evidenced an advanced level of peptidolysis. On the other hand, significant qualitative differences were observed in the free fatty acid profiles and in the sensory characteristics. Principal component analysis showed a clear separation of the fondaco and control cheeses, indicating that ripening in the natural room conferred unique sensory features to the product.

Ecological and Biological Properties of Satureja cuneifolia Ten. and Thymus spinulosus Ten.: Two Wild Officinal Species of Conservation Concern in Apulia (Italy). A Preliminary Survey.

This study evaluated the effects of ecology (plant community, topography and pedology), as well as of climate, on the composition of essential oils (EOs) from two officinal wild plant species (Lamiales) from Apulia, namely Satureja cuneifolia Ten. and Thymus spinulosus Ten. Few scientific data on their chemical composition are available, due to the fact that the first has a limited distribution range and the second is endemic of southern Italy. Results for both species, never officially used in traditional medicine and/or as spices, showed that the ecological context (from a phytosociological and ecological point of view) may influence their EO composition, and hence, yield chemotypes different from those reported in the literature. S. cuneifolia and Th. spinulosus can be considered good sources of phytochemicals as natural agents in organic agriculture due to the presence of thymol and α-pinene. Overall, the obtained trend for EOs suggests a potential use of both species as food, pharmacy, cosmetics and perfumery. Hence, their cultivation and use represent a positive step to reduce the use of synthetic chemicals and to meet the increasing demand for natural and healthier products.

Ecological and Plant Community Implication on Essential Oils Composition in Useful Wild Officinal Species: A Pilot Case Study in Apulia (Italy).

The study focused on the effects of ecology (plant communities and topographical data) on composition of essential oils (EOs) of some officinal wild plant species (Lamiales): Clinopodium suaveolens, Salvia fruticosa subsp. thomasii, Satureja montana subsp. montana, and Thymbra capitata, in different environments of Apulia (Italy). C. suaveolens and S. fruticosa subsp. thomasii are rare species of conservation interest, while S. montana subsp. montana and T. capitata, have a wide distribution and are used in traditional medicine or as spices. Results showed that the ecological context (phytosociological and ecological features) may influence the composition of EOs of the studied species. High differences in the compound composition have been found in S. montana subsp. montana, whereas minor effects were observed in C. suaveolens, S. fruticosa subsp. thomasii, and T. capitata accessions. The understanding of such aspects is necessary for providing optimal conditions to produce EOs rich in compounds known for their biological activities. The results are of great interest also for EOs producers and at the same time to improve our knowledge and valorize wild officinal plants.

Long term efficacy and safety of rivaroxaban plus cilostazol in the treatment of critical ischemia of the lower limbs in a frail, elderly patient with non valvular atrial fibrillation.

BACKGROUND: Many patients with critical lower limb ischemia are not eligible for revascularization procedures. Still, given the emerging role of both platelet and coagulation activation in the formation of arterial thrombi, they may benefit from the novel anticoagulant and antiplatelet drugs. CASE PRESENTATION: We describe the case of a male with critical lower limb ischemia complicated by older age, frailty, polymorbidity and non valvular atrial fibrillation, who was deemed as non eligible for surgery. The patient was successfully treated with the combination of rivaroxaban and cilostazol, and the clinical benefit was maintained throughout 32 months, with no occurrence of major or minor hemorrhagic or thrombotic events. CONCLUSIONS: To our knowledge, this is the first report on the efficacy and safety of such a combination therapy in critical lower limb ischemia. In a clinical setting in which alternative pharmacological approaches are urgently needed, the association of rivaroxaban and cilostazol warrants further investigations.

Protein aggregation in cooked pork products: New details on the supramolecular organization.

The molecular weight distribution of protein aggregates from raw meat and cooked pork products was assessed by size exclusion-high performance liquid chromatography (SE-HPLC). Electrophoretic analysis under reducing conditions showed that the high molecular weight SE-HPLC peak (peak 1) of the cooked products contained protein aggregates in addition to high molecular weight muscle proteins, while the second peak (peak 2) still contained aggregates and <50 kDa proteins. The protein aggregates composition was investigated by HPLC-tandem mass spectrometry. Different classes of proteins were identified and the cooked products showed a more complex composition and organization, according to the muscle structure and the technological procedures, respectively. The key role of actin in the building of the protein networks was also confirmed. The different multi-protein systems found in the cooked products suggest protein re-organization in heat-induced supramolecular structures, which might be responsible for the texture and the structural properties of the final products.

Effects of prefermentative cold soak on polyphenols and volatiles of Aglianico, Primitivo and Nero di Troia red wines.

Effectiveness of prefermentative cold soak (PCS) on polyphenols and volatiles extraction during winemaking of three red grape cultivars grown in southern Italy (Aglianico, Primitivo and Nero di Troia) was investigated. Four 200-L stainless steel horizontal rotary wine fermenters were used. The main result was that PCS improved the extraction of polyphenols and increased the antioxidant activity of wines. Extraction of proanthocyanidins was enhanced (+25%, +14% and +7% for Aglianico, Primitivo and Nero di Troia, respectively) and, consequently, the ratio flavans reactive with vanillin/proanthocyanidin was reduced, thus potentially favoring the chromatic and tannic stabilization of wines. As regards volatiles, PCS increased ester compounds at levels above their odor thresholds, potentially conferring fruity odor to wines. In conclusion, PCS could be favorably introduced in winemaking to enhance the enological potential of Aglianico, Primitivo and Nero di Troia wines.

Antioxidant activity, tocopherols and polyphenols of acornoil obtained from Quercus species grown in Algeria.

Among the unusual vegetable fats, acorn oil has interesting nutritional and functional properties. Few studies are present in literature on acorn oil characterization and antioxidant activity evaluation while no studies are present regarding the analysis of the phenolic profile. The present study aims at investigating the content of hydrophilic and lipophilic phenolic compounds and pigments, as well as antioxidant properties and quality of the oil extracted from three Quercus species grown in Algeria. Oil yield, expressed as dry weight, was in the range 7.05-8.40%. Tocopherols contents were remarkable for the three species (539-676 mg kg-1), with (ß + γ)-tocopherols being the most abundant. Oils had also significant amounts of carotenoids (42-66 mg kg-1) and similar polyphenol patterns, but with a wide quantitative variability between species (195-436 mg kg-1). Twenty phenolic compounds were detected, 12 of which were tentatively identified. All of them were hydrolysable tannins derivatives (gallotannin or ellagitannin). Oils methanolic extracts had remarkable antioxidant activity; up to 3.34 and 3.79 µmol TE g-1 oil (DPPH and ABTS test, respectively). Quercus oil is a potential source of natural antioxidants, and it could be used as a new functional oil.

First Case of 2 Synchronous Gluteal Arteries Aneurysms Treated by Endovascular Plug Embolization: Case Report and Literature Review.

INTRODUCTION: Gluteal artery aneurysms (GAAs) are rare, accounting for less than 1% of all arterial aneurysms. Most of them are post-traumatic in nature and involve the superior gluteal artery (SGA), while injuries of the inferior gluteal artery (IGA) have been reported less frequently. We report an unusual case of a patient with double saccular GAA of unknown etiology, involving both the SGA and IGA, successfully treated by endovascular embolization. CASE REPORT: A 80-year-old man referred to our hospital complaining of the progressive onset of left buttock pain and swelling exacerbated by sitting position in the last 4 months. His past medical history was positive for hypertension, prostatic adenocarcinoma treated by brachytherapy, and endocarditis diagnosed about 30 years before and treated by cardiac surgical valve replacement; no history of trauma was reported. After ultrasonography was carried out, an enhanced computed tomography (CT) scan confirmed the presence of 2 large GAAs involving both the SGA and IGA, with maximum transverse diameter of 38 and 84 mm, respectively. The patient was referred for endovascular treatment after informed consent was provided. After sequential selective catheterization of SGA and IGA, 3 Amplatzer Plugs II (St. Jude Medical, Zaventem, Belgium) were deployed inside the aneurysms. Postoperative course was uneventful as buttock pain completely disappeared on the second postoperative day. The patient was discharged to home on the third postoperative day. One-month CT scan confirmed the complete thrombosis of the aneurysms without any endoleak. CONCLUSIONS: GAAs represent a rare pathology, and for that reason, the correct timing and choice of treatment are not clearly defined. Endovascular techniques are the first step in the approach to GAAs. In case of complex anatomy, GAAs embolization by the use of vascular plugs can be successfully performed.

Ethephon As a Potential Abscission Agent for Table Grapes: Effects on Pre-Harvest Abscission, Fruit Quality, and Residue.

Some plant growth regulators, including ethephon, can stimulate abscission of mature grape berries. The stimulation of grape berry abscission reduces fruit detachment force (FDF) and promotes the development of a dry stem scar, both of which could facilitate the production of high quality stemless fresh-cut table grapes. The objective of this research was to determine how two potential abscission treatments, 1445 and 2890 mg/L ethephon, affected FDF, pre-harvest abscission, fruit quality, and ethephon residue of Thompson Seedless and Crimson Seedless grapes. Both ethephon treatments strongly induced abscission of Thompson Seedless berries causing >90% pre-harvest abscission. Lower ethephon rates, a shorter post-harvest interval, or berry retention systems such as nets, would be needed to prevent excessive pre-harvest losses. The treatments also slightly affected Thompson Seedless berry skin color, with treated fruit being darker, less uniform in color, and with a more yellow hue than non-treated fruit. Ethephon residues on Thompson Seedless grapes treated with the lower concentration of ethephon were below legal limits at harvest. Ethephon treatments also promoted abscission of Crimson Seedless berries, but pre-harvest abscission was much lower (≅49%) in Crimson Seedless compared to Thompson Seedless. Treated fruits were slightly darker than non-treated fruits, but ethephon did not affect SSC, acidity, or firmness of Crimson Seedless, and ethephon residues were below legal limits.

Production and characterisation of reduced-fat and PUFA-enriched Burrata cheese.

Burrata is an Italian fresh 'pasta filata' cheese made from cow's milk and cream that is rapidly spreading in Europe. It has very high caloric content, and a technological protocol was developed for producing a reduced-fat type and fortifying it with polyunsaturated fatty acids (PUFA) of vegetable origin. A satisfactory reduced-fat prototype was obtained by using a 14% fat cream, which was specifically developed by diluting double cream with a suspension of carob seed flour. The composition of the new cheese changed with respect to the control, but the sensory characteristics were not impaired. Moisture increased from 62·6 to 68·4%, fat on dry matter decreased from 59·1 to 34·7%, and the caloric content decreased from 1060·8 to 718 J/100 g. Proteolysis and lipolysis were not affected by the technological modifications: after 7 d storage, the electrophoretic pattern of caseins and the free fatty acids profile of experimental and control cheeses were not significantly different. Fortification of reduced-fat Burrata with PUFA was obtained by using two commercial formulates available at a compatible price with the current economic values of the cheese. The two formulates derived from flaxseeds and Carthamus tinctorius oil and allowed enrichment in C18 :3 : n3 (α-linolenic acid, ALA), and 9cis,11trans- and 10trans,12cis- conjugated linoleic acid (CLA), respectively. Fortification was easy to perform under a technical point of view, but the negative sensory impact limited fortification at a maximum of 7·0 mg g-1 fat ALA and 6·8 g-1 fat CLA.

In situ evaluation of the fruit and oil characteristics of the main Lebanese olive germplasm.

BACKGROUND: Very little information is available on the characteristics of the Lebanese olive germplasm. Therefore, the aim of this work was to evaluate the fruit and oil characteristics of the main Lebanese olive varieties (Aayrouni, Abou chawkeh, Baladi, Del and Soury) from two successive crop seasons (2010-2011). RESULTS: All of the genotypes had medium-high oil content in the fruit, indicating their suitability for oil production; Aayrouni had particularly high values. The variety Abou chawkeh also had a high pulp/pit ratio, which is a very desirable trait in table olives. For all the varieties the values of free fatty acids, peroxide values, absorbances in ultraviolet, fatty acid composition, sterol content and composition and erythrodiol + uvaol content of the oils were within the requirements of the International Olive Council's Trade Standard for extra virgin olive oil. The only exception was for the values of Δ-7-stigmastenol in 2011 in Soury and, especially, in Baladi, which were higher than 0.5%. In some cases, stearic and arachidic acids fluctuated around the maximum values allowed. CONCLUSION: The findings of this study provide a first picture of the main characteristics of olives and oils currently produced in Lebanon. © 2015 Society of Chemical Industry.

Phenols and antioxidant activity in vitro and in vivo of aqueous extracts obtained by ultrasound-assisted extraction from artichoke by-products.

Artichoke by-products are rich in phenolic compounds although they represent a waste for the food industry. This paper examines the application of ultrasound-assisted extraction (UAE) for obtaining organic solvent-free extracts rich in nutraceuticals from artichoke scraps. Application of ultrasounds for 60 minutes on test samples, using water as a solvent, improved recovery of phenolic substances compared with untreated samples. Among the phenols detected by high performance liquid chromatography, 5-O-caffeoylquinic and 1,5-di-O-caffeoylquinic acids were identified. In vivo treatments of tobacco BY-2 cells with ultrasonic extracts consistently enhanced their antioxidant power, making the cells more resistant to heat stress. UAE applied to artichoke by-products, using water as a solvent, appears to be a powerful eco-friendly technique that can provide extracts rich in nutraceuticals and turn waste products into resources. The extracts could be advantageously utilized in the food industry to produce functional foods.

Antifungal activity of Meyerozyma guilliermondii: identification of active compounds synthesized during dough fermentation and their effect on long-term storage of wheat bread.

Preliminarily, 146 strains of yeasts were screened for the antifungal activity toward the indicator Penicillium roqueforti DPPMAF1. The strain Meyerozyma guilliermondii LCF1353 was selected and used for dough fermentation. The water/salt soluble extract of the dough was analyzed by HPLC and GC/MS-SPME. The synthesis of the extracellular cell wall-degrading enzyme ß-1,3-glucanase and ethyl-acetate was shown. The effect on conidia germination mainly suggested a fungistatic activity. M. guilliermondii LCF1353 was used as starter for dough fermentation in combination with Wickeramomyces anomalus 1695 and Lactobacillus plantarum 1A7, which were previously selected for antifungal activity. The growth of the strains was monitored by plate count and molecular techniques, and competitive or antagonistic interactions among them were excluded. Bread started with the combination of M. guilliermondii LCF1353, W. anomalus LCF1695 and L. plantarum 1A7 showed a more prolonged shelf life compared to the other breads. Fungal growth was delayed at least until 14 days of storage, under conditions of high artificial inoculum. The bread manufactured with the above combination showed good chemical and textural characteristics and, as shown by sensory analysis, it was appreciated for elasticity, color and taste.

Study of a family in the province of Matera presenting with glucose-6-phosphate dehydrogenase deficiency and Gilbert's syndrome.

Glucose-6-phosphate dehydrogenase (G6PD) deficiency, a recessive X-linked trait, is the most common enzyme deficiency in the world. The most devastating clinical consequence of this deficit is severe neonatal jaundice, which results in sensorineural deficit, and severe haemolytic anemia. However, patients may be asymptomatic. The most common clinical sign is hyperbilirubinemia (h↑), that is also related to Gilbert's syndrome, a condition associated with the promoter polymorphism of the UDP-glucuronosyltransferase 1 (UGT1A1) gene. The aim of this study was to underline (as is usually done by DNA molecular analysis) to detect and to clarify the genetic deficiency that is the reason of the disorder in question. In this study, different techniques were applied to analyse a family of four individuals presenting with hyperbilirubinemia: bilirubinic dosage, electrophoresis and enzymatic activity dosage of G6PD; molecular analysis of the UGT1A promoter to detect a thymine-adenine (TA) insertion, that causes the [A(TA)7TAA] mutation. The results showed that in certain cases, the presence of hyperbilirubinemia is not only associated with G6PD deficiency, but may be caused by the co-presence of a mutation in the UGTA1 promoter related to Gilbert's syndrome. As being affected by these two conditions predisposes to adverse effects towards certain drug treatments, it is advisable to study the UGTA1 gene before prescribing drugs for specific antineoplastic or retroviral treatment. We emphasize that investigating both the UGT1A gene and G6PD activity is the most reliable way to make a correct differential diagnosis.