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Background: Diagnosis and treatment of small and isolated lung nodules remain challenging issues. Purpose: The aim of this article is to report the technique of real-time navigation using holographic reconstruction technology combined with a robot assisted thoracic surgery (RATS) platform for lung resection in patients with  small deep nodules . Research Design: The pre-surgery 3D planning was based on the chest CT scan. The reconstruction was uploaded to a head-mounted display for real-time navigation during mini invasive robot assisted surgery performed with an open console platform. We evaluated this technique with the success rate of diagnosis, the operative time and the post-operative course. Study Sample: This technique was performed in 6 patients (4 female, mean age 65 years) to date. Results: The precision of the head-mounted display based localization system was effective in all cases without the need of open conversion. The mean diameter of the nodules was 8 mm (6-9). The diagnosis was a lung cancer (n = 5) and tuberculoma (n = 1). The mean operative time was 125 min (100-145). The mean hospital stay was 2.5 days (1-3). Conclusions: In conclusion, the intraoperative navigation using the 3D holographic assistance was an helpful tool for mini invasive RATS lung segmentectomy without the need of preoperative localization., Competing Interests: Declaration of Conflicting InterestsThe author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

In the present study, we assessed the antioxidant and antileishmanial potential from fresh leaves of Ballota (B.) hirsuta essential oil (EO). The GC-MS analysis of B. hirsuta EO revealed that spathulenol and germacrene D were the main components accounting for 26.03% and 19.64% of the total EO, respectively. B. hirsuta EO possesses moderate antioxidant activity, both in neutralizing DPPH radicals and in inhibiting β-carotene bleaching. In addition, it exhibits both high antileishmanial activity and selectivity towards the promastigote and amastigote forms. Specifically, B. hirsuta EO showed an IC 50 value of 20.78 µg/mL and 23.62 µg/mL, against the promastigote and amastigote forms of L. infantum, respectively. It also demonstrated an IC 50 value of 22.39 and 25.76 µg/mL, against the promastigote and amastigote forms of L. major, respectively. However, it exhibited moderate cytotoxicity, with a selectivity index below 10. The investigation of the molecular mechanism of action revealed that B. hirsuta EO inhibited only the sterol pathway, including CYP51 gene expression. Additionally, in silico analysis indicated that the main compounds of B. hirsuta EO, germacrene and spathulenol, exhibited excellent affinity energy against Leishmania enzymes trypanothione reductase (TryR) and trypanothione synthase (TryS). This denotes the potential of these compounds as promising agents to control leishmaniasis., (© 2024. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

Combination therapy at appropriately suitable doses presents a promising alternative to monotherapeutic drugs. In this study, Cinnamomum verum and Syzygium aromaticum essential oils and their major compounds have exhibited substantial leishmaniacidal potential against both promastigote and amastigote forms of Leishmania (L.) major . However, they displayed high cytotoxicity against Raw264.7 macrophage cells. Interestingly, when combined with each other or with amphotericin B, they demonstrated a synergistic effect (FIC<0.5) with low cytotoxicity. These combinations are able to modulate the production of nitric oxide (NO) by macrophages. Notably, the combination of S. aromaticum Essential oil with amphotericin B stimulates macrophage cells by increasing NO production to eliminate leishmanial parasites. Furthermore, investigation of the molecular mechanism of action of these synergistic combinations reveals potent inhibition of the sterol pathway through the inhibition of the CYP51 gene expression. The findings suggest that combination therapy may offer significant therapeutic benefits in both food and pharmaceutical fields.

Antibiotic resistance of the Gram-negative bacterium Pseudomonas aeruginosa and its ability to form biofilm through the Quorum Sensing (QS) mechanism are important challenges in the control of infections caused by this pathogen. The extract of Myrtus communis (myrtle) showed strong anti-QS effect on C hromobacterium . violaceum 6267 by inhibiting 80 % of the production of violacein pigment at a sub-MIC concentration of 1/8 (31.25 μg/mL). In addition, the extract exhibited an inhibitory effect on virulence factors of P. aeruginosa PAO1 at half MIC (125 μg/mL), significantly reducing the formation of biofilms (72.02 %), the swarming activity (75 %), and the production of protease (61.83 %) and pyocyanin (97 %). The active fraction also downregulated the expression of selected regulatory genes involved in the biofilm formation and QS in the P. aeruginosa PAO1 strain. These genes included the autoinducer synthase genes ( lasI and rhlI ), the genes involved in the expression of their corresponding receptors ( lasR and rhlR ), and the pqsA genes. The analysis of the active fraction by HPLC/UV/MS and NMR allowed the identification of three phenolic compounds, 3,5-di- O -galloylquinic acid, myricetin 3- O -α-l-rhamnopyranoside (myricitrin), and myricetin 3- O -(2″- O -galloyl)-ß-d-galactopyranoside. In silico studies showed that 3,5-di- O -galloylquinic acid, with an affinity score of -9.20 kcal/mol, had the highest affinity to the active site of the CviR protein (3QP8), a QS receptor from C. violaceum . Additionally, myricetin 3- O -α-l-rhamnopyranoside (myricitrin) and myricetin 3- O -(2″- O -galloyl)-ß-d-galactopyranoside interact to a lesser extent with 3QP8. In conclusion, this study contributed significantly to the discovery of new QS inhibitors from M. communis leaves against resistant Gram-negative pathogens., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Carlos Jiméenez reports financial support was provided by Spanish State Agency for Research. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors.)

The present study aimed to evaluate the leishmanicidal potential of the essential oil (EO) of Micromeria ( M. ) nervosa and to investigate its molecular mechanism of action by qPCR. Furthermore, in silicointeraction study of the major M. nervosa EO compounds with the enzyme cytochrome P450 sterol 14α-demethylase (CYP51) was also performed. M. nervosa EO was analyzed by gas chromatography-mass spectrometry (GC-MS). Results showed that α-pinene (26.44%), t -cadinol (26.27%), caryophyllene Oxide (7.73 ± 1.04%), and α-Cadinene (3.79 ± 0.12%) are the major compounds of M. nervosa EO. However, limited antioxidant activity was observed, as this EO was ineffective in neutralizing DPPH free radicals and in inhibiting β-carotene bleaching. Interestingly, it displayed effective leishmanicidal potential against promastigote (IC 50 of 6.79 and 5.25 μg/mL) and amastigote (IC 50 of 8.04 and 7.32 μg/mL) forms of leishmania (L.) infantum and L. major , respectively. Molecular mechanism investigation showed that M. nervosa EO displayed potent inhibition on the thiol regulatory pathway. Furthermore, a docking study of the main components of the EO with cytochrome P450 sterol 14α-demethylase (CYP51) enzyme revealed that t -cadinol exhibited the best binding energy values (-7.5 kcal/mol), followed by α-cadinene (-7.3 kcal/mol) and caryophyllene oxide (-7 kcal/mol). These values were notably higher than that of the conventional drug fluconazole showing weaker binding energy (-6.9 kcal/mol). These results suggest that M. nervosa EO could serve as a potent and promising candidate for the development of alternative antileishmanial agent in the treatment of leishmaniasis.

This study aimed to assess the antileishmanial activity of biomolecules obtained from Olea europaea L. leaves and twigs recovered from eight Tunisian cultivars. The extraction was first carried out with 80% methanol, and then the obtained extract was fractionated using three solvents of increasing polarity: cyclohexane (CHX), dichloromethane (DCM) and ethyl acetate (EtOAc). The antileishmanial activity was determined against leishmanial strains responsible for cutaneous, visceral, and mucocutaneous leishmaniasis. The cyclohexane fraction of the leaves of cv. Chemlali from the region of Sidi-Bouzid exhibited the strongest leishmanicidal activity against all the tested leishmanial strains. The inhibition concentrations (IC 50 ) were 16.5, 14.5, and 7.4 μg mL -1 for Leishmania mexicana (cutaneous), Leishmania braziliensis (mucocutaneous), and Leishmania donovani (visceral), respectively. Interestingly, low cytotoxicity was observed on THP-1 cells with selective indexes (SI) ranging from 22.8 to 50.5. HPLC-HRMS and full-house NMR analysis allowed the identification of three triterpenic compounds, oleanolic acid (IC 50  = 64.1 μg mL -1 ), erythrodiol (IC 50  = 52.0 µg mL -1 ), and uvaol (IC 50  = 53.8 μg mL -1 ). Antileishmanial activity of uvaol and oleanolic acid has been previously reported. However, this work constitutes the first report of the antileishmanial activity of erythrodiol which showed combinatorial interaction with uvaol (IC 50  = 26.1 μg mL -1 ) against Leishmania tropica. The mixture of the three compounds, as major ones, exhibited an enhanced activity against Leishmania tropica (IC 50  = 16.3 µg mL -1 ) compared to erythrodiol alone or the combination of uvaol and erythrodiol. This finding is of great importance and needs further investigation., Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-023-03825-3., Competing Interests: Conflict of interestThe authors have no competing interests to declare that are relevant to the content of this article., (© King Abdulaziz City for Science and Technology 2023. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.)

Aim: The present study aimed to investigate a novel antifungal compound produced by Streptomyces blastmyceticus S108 strain. Its effectiveness against clinical isolates of Candida species and its synergistic effect with conventional antifungal drugs were assessed, and its molecular mechanism of action was further studied against Candida albicans., Methods and Results: A newly isolated strain from Tunisian soil, S. blastmyceticus S108, showed significant antifungal activity against Candida species by well diffusion method. The butanolic extract of S108 strain supernatant exhibited the best anti-Candida activity with a minimal inhibitory concentration (MIC) value of 250 μg ml-1, determined by the microdilution method. The bio-guided purification steps of the butanolic extract were performed by chromatographic techniques. Among the fractions obtained, F13 demonstrated the highest level of activity, displaying a MIC of 31.25 μg ml-1. Gas chromatography-mass spectrometry and electrospray ionization mass spectrometry analyses of this fraction (F13) revealed the glycolipidic nature of the active molecule with a molecular weight of 685.6 m/z. This antifungal metabolite remained stable to physicochemical changes and did not show hemolytic activity even at 4MIC corresponding to 125 µg ml-1 toward human erythrocytes. Besides, the glycolipid compound was combined with 5-flucytosine and showed a high synergistic effect with a fractional inhibitory concentration index value 0.14 against C. albicans ATCC 10231. This combination resulted in a decrease of MIC values of 5-flucytosine and the glycolipid-like compound by 8- and 64-fold, respectively. The examination of gene expression in treated C. albicans cells by quantitative polymerase chain reaction (qPCR) revealed that the active compound tested alone or in combination with 5-flucytosine blocks the ergosterol biosynthesis pathway by downregulating the expression of ERG1, ERG3, ERG5, ERG11, and ERG25 genes., Conclusion and Impact of the Study: The new glycolipid-like compound, produced by Streptomyces S108 isolate, could be a promising drug for medical use against pathogenic Candida isolates., (© The Author(s) 2023. Published by Oxford University Press on behalf of Applied Microbiology International.)

Current study aims to evaluate the effect of essential oil on the conservation of chicken fillets. Following a screening, Tetraclinis articulata was selected for its antimicrobial ( Enterococcus feacalis ATCC 29212, MIC < 0.031 mg/mL) and antioxidant (IC 50  = 1000 µg/mL) potentials with no cytotoxicity effect towards murine macrophage cells. The treatment of chicken fillets, especially with 200 ppm of T. articulata essential oil per 100 g of product reduced significantly ( p   <   0.05 ) lipid oxidation during 12 days of refrigerated storage. Microbial flora charges decreased significantly ( p   <   0.05 ) with a rate of 50.31% for 200 ppm of essential oil from the 6 th day of storage. The acidity of treated fillets was lower than control samples (1.3 g/kg) from the 3 rd day of storage. In addition, a clear effect was recorded by reducing water losses during cooking under the tested doses of essential oil. Withall, findings encouraged the use of T. articulata to extend poultry meat product shelf life.

Nanoencapsulation is widely considered as a highly effective strategy to enhance essential oils' (EO) stability by protecting them from oxidative deterioration and evaporation. The present study aims to optimize and characterize an efficient technique for encapsulating Cinnamomum (C.) verum essential oil into chitosan nanoparticles using response surface methodology (RSM). Moreover, the optimized C. verum EO nanoparticle was investigated for its antibacterial (against Gram-positive and Gram-negative bacteria), antifungal (against Candida albicans ), and antiparasitic activity (against Leishmania parasites). Five parameters were investigated using a Plackett-Burman and Box-Behnken statistical design: the chitosan molecular weight, TPP concentration, C. verum EO/chitosan ratio, mixing method, and the duration of the reaction. Encapsulation efficiency and anti-candida activity were considered as responses. The antibacterial, anticandidal, and anti-leishmanial activities were also assessed using a standard micro-broth dilution assay and the cytotoxicity assay was assessed against the macrophage cell line RAW 264.7. The optimized nanoparticles were characterized using Fourier transform infrared spectroscopy, Zeta potential, and scanning electron microscopy. The study results indicated that under optimal conditions, the nanoencapsulation of C. verum EO into chitosan nanoparticles resulted in an encapsulation efficiency of 92.58%, with a regular distribution, a nanoparticle size of 480 ± 14.55 nm, and a favorable Zeta potential of 35.64 ± 1.37 mV. The optimized C. verum EO/chitosan nanoparticles showed strong antifungal activity against C. albicans pathogens (CMI = 125 µg mL -1 ), notable antibacterial activity against both Gram-positive and Gram-negative bacteria (ranging from 125 to 250 µg mL -1 ), high leishmanicidal potential against the promastigotes form of L. tropica and L. major (IC50 = 10.47 and 15.09 µg mL -1 , respectively), and a four-fold cytotoxicity reduction compared to non-encapsulated essential oil. These results suggest that C. verum EO-loaded chitosan nanoparticles could be a promising delivery system for the treatment of cutaneous Candida albicans infections.

The present study investigated the antioxidant, antibacterial, antiviral and anti-inflammatory activities of different aerial parts (flowers, leaves and seeds) of Datura stramonium . The plant material was extracted with 80% methanol for about 24 h. The sensitivity to microorganisms analysis was performed by the microdilution technique. Antioxidant tests were performed by scavenging the DPPH and ABTS radicals, and by FRAP assay. Anti-inflammatory activity was evaluated through the inhibition of nitric oxide production in activated macrophage RAW 264.7 cells. Cell viability was assessed with an MTT assay. Results show that the flower extract revealed a powerful antimicrobial capacity against Gram-positive bacteria and strong antioxidant and anti-inflammatory activities. No significant cytotoxicity to activated macrophages was recorded. High resolution electrospray ionization mass spectrometry and nuclear magnetic resonance analysis identified two molecules with important anti-inflammatory effects: 12α-hydroxydaturametelin B and daturametelin B. Molecular docking analysis with both pro-inflammatory agents tumor necrosis factor alpha and interleukin-6 revealed that both compounds showed good binding features with the selected target proteins. Our results suggest that D. stramonium flower is a promising source of compounds with potential antioxidant, antibacterial, and anti-inflammatory activities. Isolated withanolide steroidal lactones from D. stramonium flower extract with promising anti-inflammatory activity have therapeutic potential against inflammatory disorders.

Purpose: Leishmaniasis is a parasitic disease found in tropical areas, and it affects up to 12 million individuals globally. Chemotherapies now available include drawbacks such as toxicity, high cost, and parasite resistance. This work aimed to evaluate the antileishmanial properties of essential oils (EOs) extracted from aerial parts of Cupressus sempervirens (C. sempervirens), Tetraclinis articulata (T. articulata), and Pistacia lentiscus (P. lentiscus) trees., Methods: The EOs were obtained by hydro-distillation, and chemical composition was determined by gas chromatography coupled to mass spectrometry at three phenological stages. The EOs were evaluated in vitro for antileishmanial activities against Leishmania major (L. major) and Leishmania infantum (L. infantum). The cytotoxicity effect was also tested against murine macrophagic cells (Raw264.7 lines)., Results: Results showed that P. lentiscus and T. articulata EOs presented low and moderate antileishmanial activity against L. infantum and L. major. However, C. sempervirens EO from the fructification stage gave an important selectivity index (23.89 and 18.96 against L. infantum and L. major, respectively). This activity was more interesting than those of amphotericin chemical drugs. Antileishmanial activity for this EO was highly correlated with germacrene D content (r = 1.00). This compound presented a SI equal to 13.34 and 10.38 for the two strains. According to the Principal Component Analysis (PCA), the distribution of the three phenological stages proved that the chemical composition of the EOs affected the antileishmanial activity. PCA revealed that SI was positively correlated with α-pinene, germacrene D and the sesquiterpene hydrocarbon class. Cupressus sempervirens EO can provide a source of germacrene D that can be used as a new alternative to chemical drugs for the treatment of antileishmanial diseases., Conclusion: C. sempervirens EO seemed to be a highly active antileishmanial agent and a natural alternative for chemical drugs to treat several leishmanial strains., (© 2023. The Author(s) under exclusive licence to Witold Stefański Institute of Parasitology, Polish Academy of Sciences.)

Aims: This study aimed to determine the antibacterial and antileishmanial potential of Micromeria nervosa extracts. The identification of the antileishmanial compound and the study of its molecular mechanism of action have also been undertaken., Methods and Results: Ethanol extract showed high polyphenol content and diethyl ether extract exhibited high DPPH scavenging and low beta-carotene bleaching activity (IC50 = 13.04 ± 0.99 and 200.18 ± 3.32 μg mL-1, respectively). However, diethyl ether extract displayed high antibacterial activity against Gram-positive strains including methicillin-resistant Staphylococcus aureus (MIC = 31.25 μg mL-1), Staph. aureus ATCC6538 (MIC = 62.5 μg mL-1), and Listeria monocytogenes ATCC 19115 (MIC = 125 μg mL-1), as well as high antileishmanial activity against the promastigote forms of L. infantum and L. major (IC50 = 11.45 and 14.53 μg mL-1, respectively). The active compound was purified using bioassay-guided fractionation and thin layer chromatography, and identified as ursolic acid using high-performance liquid chromatography coupled with a photodiode array and mass spectrometry. The purified compound was strongly inhibitory against the promastigote and amastigote forms of L. infantum and L. major (IC50 = 5.87 and 6.95 μg mL-1 versus 9.56 and 10. 68 μg mL-1, respectively) without overt cytotoxicity against Raw 264.7 macrophage cells (SI = 13.53 and 11.43, respectively). The commercial compound (ursolic acid) showed similar activity against amastigotes and promastigotes forms of L. infantum and L. major. Moreover, its molecular mode of action against leishmaniasis seems to involve the expression of the ODC and SPS genes involved in thiol pathway., Conclusion: Extracts of M. nervosa can be considered as a potential alternative to antimicrobial and antileishmanial drugs., (© The Author(s) 2023. Published by Oxford University Press on behalf of Applied Microbiology International.)

To explore a novel kind of green composite material having excellent antibacterial, antifungal ability and specific-targeting capability for pharmaceutical uses, a novel kind of bio-composite was prepared using sodium purified clay as carrier of Caraway essential oil (CEO). Gas chromatography-mass spectroscopy (GC-MS) analyses of CEO reveals that Carvone (68.30%) and Limonene (22.54%) are the two major components with a minimum inhibitory concentration (MIC) value equal to 125 mg/mL against Staphylococcus (S) aureus bacteria and Candida albicans fungi. Clay from Zaghouan was purified and characterized by X-ray Photoelectron Spectroscopy (XPS), X-ray diffraction (XRD), Fourier transformed infrared spectroscopy (FT-IR) and N 2 adsorption-desorption (BET method). Results obtained by chromatograph equipped with a flame ionization detector (GC-FID) show that the concentration of 130 mg/mL of essential oil and 5 h of contact with the purified clay are the optimal conditions for the bio-hybrid formation. The pseudo-second-order model can describe the kinetic study of the adsorption of Carvone and Limonene on sodium montmorillonite, and the adsorption isotherms have been established to the Langmuir type. Limonene registers a maximum adsorption value equal to 3.05 mg/g of clay however Carvone register the higher amount of adsorption (19.98 mg/g) according to its polarity and the abundance of this compound in the crude CEO. X-ray diffraction, Fourier transformed infrared spectroscopy, elemental analyses (CHN) and X-ray fluorescence characterization valid the success adsorption of CEO in sodium montmorillonite surface. The purified clay/CEO hybrid (purified clay/CEO) combined the advantages of both the clay and the essential oil used in exerting the antibacterial and antifungal activity, and thus, the composite has a double antibacterial and antifungal activity compared to the separately uses of inactive clay and CEO, suggesting the great potential application in pharmaceutical treatments., (© 2022. The Author(s).)

Biofilm formation of the opportunistic pathogen Pseudomonas (P). aeruginosa is one of the major global challenges to control nosocomial infections due to their high resistance to antimicrobials and host defense mechanisms. The present study aimed to assess the antibacterial and the antibiofilm activities of Peganum (P). harmala seed extract against multidrug-resistant P. aeruginosa isolates. Chemical identification of the active compound and determination of its molecular mechanism of action were also investigated. Results showed that P. harmala n-butanol "n-BuOH" extract exhibited antibacterial activity against multidrug-resistant P. aeruginosa isolates. This extract was even more active than conventional antibiotics cefazolin and vaamox when tested against three P. aeruginosa multidrug-resistant isolates. In addition, P. harmala n-BuOH extract exhibited potent bactericidal activity against PAO1 strain at MIC value corresponding to 500 µg/mL and attained 100% killing effect at 24 h of incubation. Furthermore, P. harmala n-BuOH extract showed an antibiofilm activity against P. aeruginosa PAO1 and exhibited 80.43% inhibition at sub-inhibitory concentration. The extract also eradicated 83.99% of the biofilm-forming bacteria. The active compound was identified by gas chromatography-mass spectrometry as an indole alkaloid harmaline. Transcriptomic analysis showed complete inhibition of the biofilm-related gene pilA when PAO1 cells were treated with harmaline. Our results revealed that P. harmala seed extract and its active compound harmaline could be considered as a candidate for a new treatment of multidrug-resistant P. aeruginosa pathogens-associated biofilm infections., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

The aim of this study was to investigate antimicrobial and antioxidant activities of different fractions obtained from edible Tunisian Ziziphus Lotus leaves of Tozeur region. Different organic extracts were tested: cyclohexane, dichloromethane, ethyl acetate, n-butanol and water. Bio-guided fractionation revealed that dichloromethane fraction is the most active against S. aureus and Methicillin-resistant S. aureus strains. Moreover, this fraction showed the highest antileishmanial activity with IC 50 values of 20.55 ± 0.34 μg/mL and 15.37 ± 0.17 μg/mL against L. major and L. infantum, respectively. The potentialities of antibacterial and leishmanicidal activities found in dichloromethane could be explained by the presence of major flavonoids such as catechin, rutin and luteolin 7-O-glucoside as revealed by HPLC system. The observed moderate antifungal activity, which was only given by butanolic fraction against pathogen fungi, may be attributed to the presence of chlorogenic acid. Furthermore, dichloromethane and butanolic fraction showed a good DPPH (2,2-diphenyl-1-picryl hydrazyl) scavenging activity and Ferric reducing power. These results suggest that Ziziphus lotus leaf fractions might be used as antioxidant and antimicrobialagent., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Introduction and Importance: Primary adenoid cystic carcinoma (ACC) of the lung is extremely rare. This tumor can be asymptomatic or have non characteristics symptoms, and the diagnosis is often late. The treatment of choice is surgery when it's possible., Case Presentation: We herein report the case of a young patient with ACC of the left main bronchus. He had dyspnea and chest pain for 6 months. Complete atelectasis of the left lung was found on the chest x-ray. Bronchoscopy showed a tumor obstructing the LMB and invading the carina. The CT scan revealed a 5 cm tumor obstructing the left main bronchus (LMB) with extension to the carina and thoracic trachea. The extension assessment was without abnormalities. The treatment was surgical. A left carinal pneumonectomy by double lateral thoracotomy was performed. The postoperative results were satisfactory. There was no recurrence with a follow-up of 2 years., Clinical Discussion: The therapeutic management of ACC is essentially based on surgical resection, which should be as radical as possible. However, complete resection is often difficult given the infiltrating nature of the tumor., Conclusion: Sleeve pneumonectomy with carinal resection is a curative option for patients with ACC of the main bronchi and carina that require expertise of the surgeons., (Copyright © 2021. Published by Elsevier Ltd.)

Cyclic lipopeptides produced by Bacillus species exhibit interesting therapeutic potential. However, their clinical use remains limited due to their low stability, undesirable interactions with host macromolecules, and their potential toxicity to mammalian cells. The present work aims to develop suitable lipopeptide-loaded chitosan nanoparticles with improved biological properties and reduced toxicity. Surfactin and bacillomycin D lipopeptides produced by Bacillus amyloliquefaciens B84 strain were loaded onto chitosan nanoparticles by ionotropic gelation process. Nanoformulated lipopeptides exhibit an average size of 569 nm, a zeta potential range of 38.8 mV, and encapsulation efficiency (EE) of 85.58%. Treatment of Candida (C.) albicans cells with encapsulated lipopeptides induced anti-adhesive activity of 81.17% and decreased cell surface hydrophobicity (CSH) by 25.53% at 2000 µg/mL. Nanoformulated lipopeptides also induced antileishmanial activity against Leishmania (L.) major promastigote and amastigote forms at respective IC 50 values of 14.37 µg/mL and 22.45 µg/mL. Nanoencapsulated lipopeptides exerted low cytotoxicity towards human erythrocytes and Raw 264.7 macrophage cell line with respective HC 50 and LC 50 values of 770 µg/mL and 234.56 µg/mL. Nanoencapsulated lipopeptides could be used as a potential delivery system of lipopeptides to improve their anti-adhesive effect against C. albicans cells colonizing medical devices and their anti-infectious activity against leishmania., (© 2021. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Echium arenarium Guss is a Mediterranean plant traditionally used in healing skin wound and it was reported exhibiting potent antioxidant, antibacterial, and antiparasitic activities. However, antitumoral activities of this plant have not yet been explored. Here we investigated for the first time, root (EARE) and aerial part (EAAPE) extracts of E. arenarium Guss to examine cytotoxicity and apoptosis activation pathway on U266 human multiple myeloma (MM) cell line. We demonstrated that EARE and EAAPE decreased U266 cell viability in a dose dependent manner. Based on 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, EARE was significantly two times more efficient (IC50 value 41 μg/ml) than EAAPE (IC50 value 82 μg/ml) considering 48 h of treatment. Furthermore, after 24 h of exposure to 100 μg/ml of EARE or EAAPE, cell cycle showed remarkable increase in sub-G1 population and a decrease of U266 cells proportion in G1 phase. In addition, EARE increased cell percentage in S phase. Moreover, analysis revealed that EAAPE or EARE induced apoptosis of U266 cells after 24 h of treatment. Interestingly, depolarization of mitochondrial membrane potential and activation of caspase 3/7 were demonstrated in treated U266 cells. Phytochemical analysis of E. arenarium extracts showed that EARE exhibited the highest content of total phenolic content. Interestingly, six phenolic compounds were identified. Myricitrin was the major compound in EARE, followed by luteolin 7-O-glucoside, resorcinol, polydatin, Trans-hydroxycinnamic acid, and hyperoside. These findings proved that an intrinsic mitochondria-mediated apoptosis pathway probably mediated the apoptotic effects of E. arenarium Guss extracts on U266 cells, and this will suggest several action plans to treat MM., (© 2021 Wiley Periodicals LLC.)

An endophytic Bacillus amyloliquefaciens strain called C5, able to produce biosurfactant lipopeptides with a broad antibacterial activity spectrum, has been isolated from the roots of olive tree. Optimization of antibacterial activity was undertaken using grape seed flour (GSF) substrate at 0.02, 0.2, and 2% (w/v) in M9 medium. Strain C5 exhibited optimal growth and antimicrobial activity (MIC value of 60 μg/ml) when incubated in the presence of 0.2% GSF while lipopeptide production culminated at 2% GSF. Thin layer chromatography analysis of lipopeptide extract revealed the presence of at least three active spots at Rf 0.35, 0.59, and 0.72 at 0.2% GSF. Data were similar to those obtained in LB-rich medium. MALDI-TOF/MS analysis of lipopeptide extract obtained from 0.2% GSF substrate revealed the presence of surfactin and bacillomycin D. These results show that GSF could be used as a low-cost culture medium supplement for optimizing the production of biosurfactants by strain C5.

Four hundred and fifty bacteria were evaluated for antagonistic activity against bacterial soft rot of potato caused by Pectobacterium carotovorum sp strain II16. A strain Ar10 exhibiting potent antagonist activity has been identified as Bacillus amyloliquefaciens on the basis of biochemical and molecular characterization. Cell free supernatant showed a broad spectrum of antibacterial activity against human and phytopathogenic bacteria in the range of 10-60 AU/mL. Incubation of P. carotovorum cells with increasing concentrations of the antibacterial compound showed a killing rate of 94.8 and 96% at MIC and 2xMIC respectively. In addition, the antibacterial agent did not exert haemolytic activity at the active concentration and has been preliminary characterized by TLC and GC-MS as a glycolipid compound. Treatment of potato tubers with strain Ar10 for 72 h significantly reduced the severity of disease symptoms (100 and 85.05% reduction of necrosis deep / area and weight loss respectively). The same levels in disease symptoms severity was also recorded following treatment of potato tubers with cell free supernatant for 1 h. Data suggest that protection against potato soft rot disease may be related to glycolipid production by strain Ar10. The present study affords new alternatives for anti-Pectobacterium carotovorum bioactive compounds against the soft rot disease of potato., (Copyright © 2018 Elsevier GmbH. All rights reserved.)

The use of synthetic food additive and the appearance of antibiotic resistance are at the basis of important human health problems. The substitution of synthetic compounds with new natural substances extracted from plants or microorganisms is therefore the ideal solution to this scourge. The objective of this work was to evaluate the phyto-constituents (polyphenols, flavonoids and condensed tannins), and to test the biological activities (antioxidant, antibacterial and antiviral) of the Ajuga iva (L) aerial part extracts. The antioxidant activity assayed by DPPH method showed an IC 50 of 0.43 ± 0.03 mg/mL. Antibacterial activity of aqueous and hydro methalonic extracts was tested against seven pathogenic bacteria (Staphylococcus aureus, Methicillin Resistant Staphylococcus aureus (MRS), Listeria monocytogenes, Pseudomonas aeruginosa, Bacillus cereus, Escherichia coli and Salmonella enteritidis) using the diffusion method. A Thin Layer Chromatography-bioautotography-guided was performed, and the isolated antibacterial fraction was identified by CG-MS analysis. Antiviral effect of methanolic extract performed on 4 viruses: Coxsackie Virus type B-3 (CVB-3), Adenovirus type 5 (ADV-5), Respiratory Syncytial Virus type B (RSV-B) and Herpes Simplex Virus type 2 (HSV-2) showed an activity against Coxsackie Virus. As a result of this study, the aerial parts of Ajuga iva (L) extract could be used in the food, cosmetic, medical and health sectors., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

The present work was developed to evaluate the in vitro antioxidant, antibacterial, antileishmanial and cytotoxic activities of Echium arenarium (Guss) extracts, and to analyze their phytochemical composition. The highest content of total phenolic compounds was obtained in the ethyl acetate extract which showed the best DPPH scavenging activity and β-carotene bleaching inhibition (IC 50  = 1.1 and 9.94 μg/mL respectively). It also exhibited the highest antibacterial activity against Gram-positive bacteria (L. monocytogenes; S. aureus; MRSA, E. faecalis and B. cereus) and antileishmanial activity against L. major (IC 50  = 13.91 ± 0.43 μg/mL) and L. infantum (IC 50  = 9.91 ± 0.15 μg/mL). Moreover, the active extract exhibited potent antiamastigote activity (IC50 = 22.48 ± 0.14 μg/mL and 18.59 ± 0.09 μg/mL against L. major and L. infantum respectively). Cytotoxicity studies revealed low toxicity against Raw 264.7 macrophage cell line (IC50 = 145.80 ± 0.84 μg/mL, SI < 10). Luteolin-7-O-glucoside was identified as the major flavonoid component by RP-HPLC analysis. In conclusion, Echium arenarium (Guss) extract was characterized by a wide range of biological activities and could be used as a potential natural anti-infectious drug., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Cryptosporidium is an enteric parasite infecting a wide range of hosts. It has emerged as an important cause of chronic life-threatening diarrhea in humans worldwide. Several subtypes of Cryptosporidium sp. have been described to be responsible for several large outbreaks related to water contamination in developed countries. However, there is a lack of information in the genetic diversity of Cryptosporidium among human population especially in developing countries. The present study aimed to update and report the genetic diversity of human Cryptosporidium spp. at the subtype level in an urban area of Tunisia using the 18S rRNA and gp60 gene. Genotyping of 42 Cryptosporidium positive isolates from different human populations at the 18S rRNA locus has identified three Cryptosporidium species: C. hominis (n = 20), C. parvum (n = 19), C. meleagridis (n = 2) and a co-infection C. hominis/C. meleagridis (n = 1). The sub-genotyping of these isolates at the 60-kda glycoprotein (gp60) locus was possible in 40 cases. It showed the presence of three subtype families (IIa, IIb and IIc) within C. parvum, a single subtype family within C. hominis and C. meleagridis isolates (Ia and IIIb respectively). Several subtypes were implicated in different human populations with the dominance of IaA26G1R1, IIaA15G2R1, IIdA16G1R1, IIdA22G2R1 and IIIbA26G1R1 variant respectively for C. hominis, C. parvum and C. meleagridis. The distribution of Cryptosporidium isolates in urban area of Northern Tunisia was dominated by the anthroponotic transmission via C. hominis species and the IIc subtype of C. parvum. However, zoonotic transmission is still possible in this region via zoonotic subtypes of C. parvum (IIa and IId) and C. meleagridis (IIIb). Subtype diversity was higher in this area., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Background: Cryptosporidiosis represents a major public health problem. This infection, caused by a protozoan parasite of the genus Cryptosporidium, has been reported worldwide as a frequent cause of diarrhoea. In the immunocompetent host, the typical watery diarrhea can be self-limiting. However, it is severe and chronic, in the immunocompromised host and may cause death. Cryptosporidium spp. are coccidians, which complete their life cycle in both humans and animals. The two species C. hominis and C. parvum are the major cause of human infection. Compared to studies on C. hominis and C. parvum, only a few studies have developed methods to identify C. meleagridis., Aim: To develop a new real time PCR-coupled High resolution melting assay allowing the detection for C. meleagridis, in addition of the other dominant species (C. hominis and C. parvum)., Methods: The polymorphic sequence on the dihydrofolate reductase gene (DHFR) of three species was sequenced to design primers pair and establish a sensitive real-time PCR coupled to a high-resolution melting-curve (HRM) analysis method, allowing the detection of Cryptosporidium sp. and discrimination between three prevalent species in Tunisia. We analyzed a collection of 42 archived human isolates of the three studied species., Results: Real-time PCR coupled to HRM assay allowed detection of Cryptosporidium, using the new designed primers, and basing on melting profile, we can distinguish C. meleagridis species in addition to C. parvum and C. hominis., Conclusion: We developed a qPCR-HRM assay that allows Cryptosporidium genotyping. This method is sensitive and able to distinguish three Cryptosporidium species., (Copyright © 2017. Published by Elsevier Ltd.)

Cryptosporidium spp. are a major cause of gastrointestinal diseases in humans worldwide. While a single subtype of Cryptosporidium hominis has been shown to be responsible for several large outbreaks related to water contamination in developed countries, little is known about the epidemiology of C. hominis in developing countries. This study reports the first genetic characterization of C. hominis at the subtype level in several human populations in Tunisia using the gp60 gene. Eighteen isolates were identified as C. hominis by a restriction fragment length polymorphism (RFLP) analysis. The prevalence of this species in different human populations ranges from 1.53% to 13.04% with a high prevalence being reported in immunocompromised children (13.04%) followed by patients with malignent myeloma (5.5%) and HIV-infected patients (4.59%). The gp60 analysis on C. hominis isolates, performed in 14 cases, showed the presence of a single subtype family: "Ia". Different subtypes were identified within this family (A11G1R1, A12R3, A23G1R1, A26G1R1, A27G1R1, A28G1R1). The IaA26G1R1 subtype was the most dominant subtype described in this area (50%). Despite the high genetic diversity of Cryptosporidium spp, a low heterogeneity at the subtype level was observed within C. hominis circulating in Tunisia. This distribution is an indicator for intensive and stable anthroponotic cryptosporidiosis in this region. Besides, the presence of a unique genotype in 5 HIV-infected patients attending the same hospital ward suggests the possible occurrence of hospital-acquired infection and underlines the need to implement preventive measures to avoid nosocomial transmission., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

The present study aimed to investigate the anti-Candida activity of ten essential oils (EOs) and to evaluate their potential synergism with conventional drugs. The effect on secreted aspartic protease (SAP) activity and the mechanism of action were also explored. The antifungal properties of essential oils were investigated using standard micro-broth dilution assay. Only Cinnamomum verum, Thymus capitatus, Syzygium aromaticum, and Pelargonium graveolens exhibited a broad spectrum of activity against a variety of pathogenic Candida strains. Chemical composition of active essential oils was performed by gas chromatography-mass spectrometry (GC-MS). Synergistic effect was observed with the combinations C. verum/fluconazole and P. graveolens/fluconazole, with FIC value 0.37. Investigation of the mechanism of action revealed that C. verum EO reduced the quantity of ergosterol to 83%. A total inhibition was observed for the combination C. verum/fluconazole. However, P. graveolens EO may disturb the permeability barrier of the fungal cell wall. An increase of MIC values of P. graveolens EO and the combination with fluconazole was observed with osmoprotectants (sorbitol and PEG6000). Furthermore, the combination with fluconazole may affect ergosterol biosynthesis and disturb fatty acid homeostasis in C. albicans cells as the quantity of ergosterol and oleic acid was reduced to 52.33 and 72%, respectively. The combination of P. graveolens and C. verum EOs with fluconazole inhibited 78.31 and 64.72% SAP activity, respectively. To our knowledge, this is the first report underlying the mechanism of action and the inhibitory effect of SAP activity of essential oils in synergy with fluconazole. Naturally occurring phytochemicals C. verum and P. graveolens could be effective candidate to enhance the efficacy of fluconazole-based therapy of C. albicans infections.

Background: In Tunisia, detection of Plasmodium in asymptomatic individuals from endemic countries is a critical measure in national program of malaria eradication. The screening is based on microscopic examination of thick and thin blood smears. However, the performance of this diagnosis is closely related to the experience of biologist and the parasitaemia., Aim: The objective of this study was to evaluate the contribution of the PCR in the screening of malaria., Methods: This prospective study involved 260 students from malaria endemic areas who were screened for malaria between september 2011 and june 2013. Each subject had a blood sample which was examined for malaria by microscopy and nested multiplex PCR., Results: PCR detected the presence of Plasmodium in 13 blood samples (5%). While microscopy was positive only in nine cases (3.5%). The discordances involved five negative samples at microscopy and which were positive in PCR and a negative sample in PCR which was positive at microscopy. A mixed infection with Plasmodium falciparum and Plasmodium malariae was identified by PCR. For this case, microscopy diagnosed only Plasmodium falciparum specie., Conclusion: PCR is more efficient than microscopy in detecting low parasitaemia ; particularly observed in asymptomatic subjects. This technique allows to reduce asymptomatic carriage of Plasmodium and reduce the risk of a resumption of transmission of malaria in our country.

We report the first case of an imported Plasmodium ovale relapse in a Tunisian man who developed malaria three years after leaving sub- Saharan Africa. A 29-year-old Tunisian man consulted in September 2011 because of a fever, myalgia, and headache that had begun eight days earlier and persisted despite treatment with oral antibiotics. On questioning, the patient stated that he had resided three years ago for six months in Ivory Coast, where he acquired malaria. He was treated with artemether-lumefantrine. The patient said he had no recent travel to any other malaria-endemic area and had not received a blood transfusion. A first microscopy of peripheral blood smears was negative for malaria parasites. The diagnosis was established 17 days after onset of symptoms. A repeat microscopic examination of blood smears confirmed the presence of Plasmodium ovale with a parasitemia lower than 0.1%. The patient was treated with artemether lumefantrine, followed by primaquine. This case emphasizes the possibility of relapse of some plasmodial species. It highlights the importance of repeating microscopic examination of blood when the diagnosis of malaria is suspected.

Several volatile organic compounds (VOCs) producing endophyte bacteria were isolated from the leaves of olive trees and tested for their antifungal activity against several pathogenic fungi. An antagonistic strain called P2 showed 97 % of homology with Pseudomonas sp. strains on the basis of its 16S rDNA sequence and biochemical properties. P2 strain drastically inhibited the growth of Rhizoctonia solani mycelia (86 %) at 5 day-post-confrontation (dpc) and strongly reduced fungi infection on potato slices at 10(7) bacteria ml(-1) for 3 and 7 dpc. P2 strain was also positive for protease activity as well as siderophore production. Light microscopy analysis showed that treatment of R. solani mycelia with P2 strain induced thickening of the cell-wall, vesiculation of protoplasm and blockage of fungal hyphae branching. VOCs analysis using GC-MS allowed the detection of two major products with m/z of 93.9910 and 125.9630 corresponding to dimethyl disulfide and dimethyl trisulfide respectively. VOCs-producing P2 strain could be a promising agent in the protection of tuber crops against fungal diseases.

The zoonotic potential of Cryptosporidium parvum was studied in an extensive cattle farming region of northern Tunisia. Seventy fecal samples from pre-weaning calves and 403 fecal samples from children were examined by microscopy after modified Ziehl-Neelsen (MZN) staining. Positive Cryptosporidium specimens were identified at a species level using an 18S rRNA nested polymerase chain reaction (PCR) followed by an Restriction Fragment Length Polymorphism (RFLP) analysis. C. parvum isolates were subgenotyped by sequence analysis of the glycoprotein 60 (gp60) gene. Among calf samples, 14 samples were positive by MZN method. C. parvum was identified in all cases. Twelve parvum isolates (85.7%) belonged to family subtype IIa. Subtype IIaA15G2R1 was more prevalent (50%). Two C. parvum isolates corresponded to the IIdA16G1 subtype. Seven human samples were positive by MZN method. C. parvum and C. meleagridis were identified in four and three cases, respectively. Intraspecific characterization of C. parvum identified two subtypes, the IIaA15G2R1 and the IIdA16G1, also found in calves.

Background: The present study was conducted to investigate the serological survey of Toxoplasma antibodies in local.horses from three major regions: a neighbourhood of a city in the North (Sidi Thabet), a neighbourhood of a city on the coast (Monastir) and a neighbourhood of a city in the middle (Battan) of Tunisia (North of Africa)., Methods: A total of 158 serum samples were obtained from clinically healthy horses which consisted of 111 (32 female, 79 male) 2-10 years old and 47 (11 female, 36 male) older than 10 years. All of the horses were tested for antibodies to T. gondii using the Modified Agglutination Test (MAT)., Results: According to MAT results, antibodies to T. gondii were found in 28 (17.7%) of 158 sera with the titers of 1:20 in 20 horses, 1:40 in 1 horse, 1:80 in 2 horses, 1:160 in 2 horses, 1:320 in 1 horse and ≥1:640 in 2 horses. Anti-T. gondii antibodies were found in 18 (16.2%) of 111 horses (2-10 years old) and 10 (21.2%) of 47 horses (older than 10 years old). Six (13.9%) out of 43 female had anti-toxoplasma antibodies and 22 (19.1%) from 115 males remained positive., Conclusion: Statistically significant differences in age groups and genders were observed between the seropositive and seronegative horses using the Chi square X(2) test. Other statistical correlation was also reported concerning horse breed.

Background: In Tunisia, Cryptosporidium is frequently identified in diarrheic stools of children and immunocompromised patients. The infection is usually self-limited in immunocompetent populations, but can be severe and life-threatening in immunocompromised individuals. Cryptosporidiosis is well-documented in patients with the human immunodeficiency virus; however, few data are available concerning children with primary immunodeficiencies (PIDs)., Patients and Methods: A retrospective study was conducted on 5 cryptosporidiosis cases diagnosed in 11 children with PIDs. Cryptosporidium was systematically investigated when patients presented chronic diarrhea. Stool samples were examined for the parasite oocysts by modified Ziehl-Neelsen staining, and DNA was systematically extracted for a nested polymerase chain reaction (PCR). The species were identified by the analysis of restriction patterns. Epidemiological and clinicobiological data were presented for each patient., Results: All cryptosporidiosis cases presented a CMH class II deficiency syndrome. Chronic diarrhea was associated with failure to thrive in all cases. PCR provided the diagnosis in all patients, while Ziehl-Neelsen staining revealed Cryptosporidium oocysts in only 3 cases. Species identification yielded Cryptosporidium hominis in 2 cases, Cryptosporidium meleagridis in 1 case, and Cryptosporidium parvum in 1 case; a C. hominis/C. meleagridis co-infection was observed in the last case. C. hominis was isolated in children from rural areas, suggesting that the infection could have been contracted in the hospital and thus a probability of nosocomial transmission. One of the C. hominis carriers developed sclerosing cholangitis with a high parasite load., Conclusion: Cryptosporidiosis with serious clinical symptoms is observed in PID patients, particularly those with CMH class II deficiency syndrome. Early, regular, and repeated screening, improved by PCR, is recommended in this group of patients. The predominance of C. hominis, the anthropophilic species, in children from rural areas should emphasize hygiene measures in care centers where PID cases are treated., (Copyright © 2011 Elsevier Masson SAS. All rights reserved.)

Microsporidiosis and cryptosporidiosis are emerging opportunistic infections responsible for intestinal manifestations that are often severe in immunocompromised patients. A case of microsporidiosis-cryptosporidiosis coinfection is reported in an HIV-infected newborn. The patient was a 17-day-old female, exclusively breastfed and with no contact with animals. Microsporidiosis and cryptosporidiosis were diagnosed after systematic screening in stool samples using both specific staining and PCR. Two species of microsporidia, Encephalitozoon intestinalis and Enterocytozoon bieneusi, and Cryptosporidium hominis were identified. The contamination of the newborn probably resulted from direct human-to-human transmission during close contact with the mother (who had diarrhea and refused stool sampling). This report highlights the usefulness of the screening of intestinal microsporidiosis and cryptosporidiosis in HIV-infected subjects for better management., (Copyright © 2011 Elsevier Masson SAS. All rights reserved.)