Long-term therapeutic outcomes of facial arteriovenous malformations: A retrospective bi-centric study of 30 cases.

INTRODUCTION: Among vascular malformations, arteriovenous malformations (AVM) are potentially the most invasive and destructive especially when located on the face. Their management is still subject to controversy and yet no consensus exists. Our aim was to report long-term therapeutic outcomes for patients with facial AVM managed either by embolization alone or by resection with/without preoperative embolization. MATERIAL AND METHODS: A bi-centric retrospective study was carried out covering the period from 2001 to 2018 including 30 patients with a facial AVM. Outcomes were categorized as follows: with 1=controlled disease, 2=improved disease (residual, no expansion), 3=persistent or stable disease (neither improved nor worsened), and 4=recurrent or worsened disease. RESULTS: The initial treatment modality was embolization (n=5, 16.7%), surgical resection (n=16, 53.3%), and surgical resection after embolization (n=9, 30%). The follow-up period ranged from 12 to 216 months with a median of 54.9 months. Taking all treatment modalities together, disease control was achieved in 60% of the cases. Disease control was achieved in 77.8% of the cases after embolization followed by surgery, in 68.7% after surgery alone and in none of the cases after embolization alone. CONCLUSIONS: According to our results, optimal treatment is based on a combination of embolization followed by a well-conducted surgical treatment.

Implication of efflux pumps and ERG11 genes in resistance of clinical Trichosporon asahii isolates to fluconazole.

Introduction. Trichosporon asahii has been recognized as an opportunistic agent having a limited sensitivity to antifungal treatment.Hypothesis/Gap Statement. Molecular mechanisms of azole resistance have been rarely reported for Trichosproron asahii. Similar to other fungi, we hypothesized that both ERG11 gene mutation and efflux pumps genes hyper-expression were implicated.Aim. The current work aimed to study the sensitivity of clinical T. asahii isolates to different antifungal agents and to explore their resistance mechanisms by molecular methods including real-time PCR and gene sequencing.Methods. The sensitivity of T. asahii isolates to fluconazole, amphotericin B and voriconazole was estimated by the Etest method. Real-time PCR was used to measure the relative expression of Pdr11, Mdr and ERG11 genes via the ACT1 housekeeping gene. Three pairs of primers were also chosen to sequence the ERG11 gene. This exploration was followed by statistical study including the receiver operating characteristic (ROC) curve analysis to identify a relationship between gene mean expression and the sensitivity of isolates.Results. In 31 clinical isolates, the resistance frequencies were 87, 16.1 and 3.2 %, respectively, for amphotericin B, fluconazole and voriconazole. Quantitative real-time PCR demonstrated that only Mdr over-expression was significantly associated with FCZ resistance confirmed by univariate statistical study and the ROC curve analysis (P <0.05). The ERG11 sequencing revealed two mutations H380G and S381A in TN325U11 (MIC FCZ=8 µg ml-1) and H437R in TN114U09 (MIC FCZ=256 µg ml-1) in highly conserved regions (close to the haem-binding domain) but their involvement in the resistance mechanism has not yet been assigned.Conclusion. T. asahii FCZ resistance mechanisms are proven to be much more complex and gene alteration sequence and/or expression can be involved. Only Mdr gene over-expression was significantly associated with FCZ resistance and no good correlation was observed between FCZ and VCZ MIC values and relative gene expression. ERG11 sequence alteration seems to play a major role in T. asahii FCZ resistance mechanism but their involvement needs further confirmation.

Unusual case of otomycosis caused by Saksenaea vasiformis.

Saksenaea vasiformis is a species of the order Mucorales rarely reported as a cause of human mucormycosis. We report an unusual case of S. vasiformis otitis occurring in a diabetic woman after penetration of an insect in the right ear. Direct microscopic examination of the clinical sample showed hyaline and non septate hyphae belonging to the order Mucorales. Fungal identification was performed by sequencing the ITS region of the rDNA. To our knowledge, this is the first report of S. vasiformis infection in Tunisia.

Microsatellite analysis of the population structure in Rhizopus arrhizus.

OBJECTIVES: Rhizopus arrhizus is recognized as an emergent agent of superficial and invasive mucormycosis. Despite an increasing number of these infections, the molecular epidemiology of Rhizopus species has not been well studied. MATERIALS AND METHODS: In this study, 43 R. arrhizus strains (25 environmental and 18 clinical isolates) were genotyped using six novel panels of microsatellite markers. RESULTS: Upon the analysis of 43 isolates, 4-8 distinct alleles were detected for each marker. The discriminatory power for the individual markers ranged from 0·522 to 0·830. The combination of all six markers yielded 33 different haplotypes with a high degree of discrimination (0·989 D value). A four-marker combination were selected as the most parsimonious panel achieving D > 0·95. One clinical isolate and one environmental isolate shared the same genotype suggesting the possible nosocomial outbreak of mucormycosis in hospitalized patients. We have noted that the strains isolated from cutaneous mucormycosis were different from the strains isolated from rhino-orbito-cerebral mucormycosis. Then, the hypothesis of particular tropism of infectious strains for a given site is not excluded. The standardized indices of association IA and rBarD were significantly different from zero (P < 0·01), suggesting a prevailing clonal reproduction. The environmental population was significantly differentiated from clinical populations (Fst = 0·2249). CONCLUSIONS: Microsatellite typing method described in our study showed an excellent degree of discriminatory power. It is a promising tool for illuminating the molecular epidemiology of R. arrhizus species, including strain relatedness and transmission pathways.

Detection of virulence factors and antifungal susceptibility of human and avian Aspergillusflavus isolates.

Aspergillusflavus is the second leading cause of invasive and non-invasive aspergillosis. Secretion of hydrolytic enzymes is considered as a virulence factor in this species. Our work aimed to study in vitro production of some virulence factors, to evaluate the biofilm production against human and avian A. flavus isolates and to investigate the antifungal susceptibility agents. Hydrolytic enzymes, biofilm production and molecular typing were studied for 62 human and 36 avian A. flavus isolates by specific solid media and six microsatellite markers. The susceptibility to antifungal agents was evaluated for 37 human isolates. All human and avian A. flavus isolates showed positive activities of extracellular hydrolase: phospholipase, protease and hemolysin. A positive elastase activity was seen in 64.51% of human A. flavus isolates and 86.1% of avian A. flavus isolates. All A. flavus in these two populations formed biofilms. Statistical significant difference was observed for the mean phospholipase activities (P=0.025) and biofilm quantification (P=0.0001) between human and avian A. flavus isolates. The in vitro susceptibility results showed a resistance in 83.7%, 81.08% and 16.21% of A. flavus isolates respectively to amphotericin B, itraconazole and posaconazole. No association was noted between all virulence factors and the genotypes of human and avian isolates. Our study allowed us to show that human strains have a higher production of extracellular hydrolases and biofilm then avian strains. These virulence factors appear to act synergistically to contribute to the virulence of A. flavus strains. Moreover, significant correlation between virulence patterns and antifungal susceptibility profiles was observed.

Contribution of the internal transcribed spacer regions to the detection and identification of human fungal pathogens.

Fungi are morphologically and phylogenetically diverse. There identification is largely based on phenotypic methods. Thus, related species, phenotypic variants and rare species may be unidentified. So, molecular methods have been introduced for identification of pathogenic molds to overcome these problems. In this study, we report the contribution of molecular tools (PCR sequencing) to identify fungal pathogens in both clinical and environmental samples. A total of 82 mold isolates were used (50 clinical samples and 32 environmental samples). PCR and direct sequencing, targeting the internal transcribed spacer (ITS) regions, were performed. We employed comparative sequence analysis to identify molds by using the GenBank database. 89% of isolates were identified by phenotypic methods. PCR- sequencing allowed the fungal identification in all cases. The concordance between molecular and morphological identification was obtained for 33 cases (40.2%). In 36 cases (43.9%), the molecular study gave the exact species identification. PCR sequencing allowed as revising mycological identification for 13 fungi strains (15.9%). The concordance of identification at species level by phenotypic method and by sequence analysis was obtained for 28% of clinical samples and for 59% of environmental samples. The phylogenetic tree for the ITS sequences showed six different clusters that are composed of isolates belonging to the same genus or species. PCR sequencing has been shown to be useful for the detection of the presence of fungal DNA in both environmental and clinical samples. It is rapid and more sensitive for the identification of medically important fungi.

Investigations of the charge transfer phenomenon at the hybrid dye/BiVO4 interface under visible radiation.

Photocatalytic hybrid systems were realized by associating bismuth vanadate (BiVO4) nanostructured thin films with anchored organic and metal-organic complex molecules. The chosen dyes are based on indoline and azo-based moieties. Optical and photoinduced charge transfer features were investigated experimentally and analysed theoretically through the electron band alignment on the organic/inorganic interface. Quantum calculations were carried out for the studied hybrid systems by using DFT and semi-empirical approaches. The calculations were performed by implementing a cluster model applied for the nanostructures and hybrid systems. The electronic density peculiarities point out efficient charge transfer for D149 based hybrids compared to azo-based systems. The electron distribution in hybrid systems inferred from the computational analysis and their experimental probing using Kelvin Force Microscopy (KFM) maps the way to understanding the photoinduced charge transfer occurring at the interfaces between organic dyes and an inorganic photocatalyst. The presented approach helps to predict suitable photoactive hybrid materials leading to efficient photocatalytic devices.

Prevalence of free living amoeba in the domestic waters reservoirs in Sfax, Tunisia.

Free living amoebas (FLA) are opportunistic pathogen found in different water sources in the environment. The aim of this study was to investigate the prevalence of free living amoeba in different samples of domestic water reserves (DWR) in Sfax region from Tunisia. It was a prospective study dealing with 486 water samples collected from different DWR. After filtration through a cellulose acetate membrane samples were cultured on non-nutrient agar and the FLA were detected and strained with Giesma, Trichrome and red nuclear stain for morphological and morphotypic studies. FLA were found in 62% of samples. The Acanthopodial morphotype was detected in 43%, Polytactic (38%), Monotactic (28%), Fan-shaped (17%), Rugose (11%), Dactilopodial (10%) and Eruptive (9%). These results demonstrate that domestic water reserves are a significant source of the FLA and maintenance of DWR is recommended.

Epidemiology of antifungal susceptibility: Review of literature.

Fungal infections are a major cause of morbidity and mortality despite the latest developments of diagnostic tools and therapeutic options. Early initiation of the appropriate antifungal therapy has been demonstrated to have a direct impact on the patient's outcome. Antifungal susceptibility testing methods are available to detect antifungal resistance and to determine the best treatment for a specific fungus. American and European standards have been developed, as well as equivalent commercial systems, which are more appropriate for clinical laboratories. These studies have allowed the development of interpretative breakpoints against the most frequent agents of fungal infections in the world. Surveillance of antifungal susceptibility patterns can provide the local drug resistance data to the clinicians, which can further aid better management of patients. Antifungal susceptibility tests have become essential tools to identify resistance to antifungals, to know the local and global disease epidemiology and to guide the treatment of fungal diseases. The distribution of species and the prevalence of antifungal resistance in fungi isolates varied among different areas. Here we summarize the epidemiology of antifungal susceptibility pattern of different fungal species.

Orbital cellulitis secondary to a fungal sinusitis caused by Scopulariopsis: The first case in Tunisia.

We report a case of invasive fungal sinusitis caused by Scopulariopsis in 57 year-old man who had recurrence of orbital cellulitis. CT-scan and magnetic resonance imaging found an orbital cellulitis associated to a left frontal sinusitis with bone erosion and calcification. Patient was treated by surgical debridement and voriconazole. Culture of excised tissue was positive for Scopulariopsis.

Postpartum hemorrhage: new insights for definition and diagnosis.

The current definition of is inadequate for early recognition of this important cause of maternal death that is responsible for >80,000 deaths worldwide in 2015. A stronger definition of postpartum hemorrhage should include both blood loss and clinical signs of cardiovascular changes after delivery, which would help providers to identify postpartum hemorrhage more promptly and accurately. Along with the amount of blood loss, clinical signs, and specifically the shock index (heart rate divided by systolic blood pressure) appear to aid in more accurate diagnosis of postpartum hemorrhage.

Isolation and molecular identification of Acanthamoeba spp from oasis water in Tunisia.

In the southern Tunisia Oasis, we conducted 211 water with drawals from various water traffic sites. This water is used for agriculture, swimming or various other human activities. Acanthamoeba genus was detected in 82% of collected samples. Sequencing of the amplification products with primers P892C/P892 has allowed us to detect genotypic variation with predominance of T4 genotype (51%) and presence of the genotypes T14, T5, T3, T16, T15, T10, T11, T9 and T7. They T4, T3, T5, T15, T11 and T10 genotypes have a high potential for pathogenicity and a very high degree of virulence due to their production of serine proteases and extracellular cysteine enzymes involved in tissue degradation of the host. T4 genotype was the most abundant in the environment as well as in infections caused by Acanthamoeba spp. T5 genotype was ranked second and T3 genotype was less abundant in the environment and its pathogenicity is discussed. Acanthamoeba strains with the genotypes T16, T9 and T7 were considered non pathogenic. In fact, they have been isolated only from the environment. However, for these strains, their role as a reservoir can be a real risk to human health.

Environmental and molecular study of fungal flora in asthmatic patients.

The aim of the present study was to investigate the epidemiological and fungal environmental profile in asthmatic patients. We conducted a prospective study involving 49 patients with allergic asthma. One hundred and forty-five clinical samples and 289 environmental samples were performed. Only 30 patients accepted to participate to the environmental study at their home. For specific IgE antibodies, ELISA assay was conducted for 21 patients. Molecular ITS sequencing was performed for 37 isolates. The frequency of attacks was significantly associated with the seasonality, which was closely related to climate (P=0.024), exposure to animals (cats, P=0.025), plants (olive, P=0.018), physical effort (P=0.04) and the number of permanent occupants in house (>6) (P=0.026). Fungal contaminants were detected from 78.6% of biological samples and 97.8% of environmental samples. Antibodies corresponding to the studied allergens were detected in 10 patients (10/21). PCR sequencing allowed as rectified morphological identification for 27.02% (10/37) strains of Aspergillus. The allergy in molds is an indisputable reality that is necessary to look for in front of any severe asthma. So, it is important to establish clearly a relationship between exposure to fungi and health disorders in order to set up specific and effective preventive measures.

Influence of the 2014-2015 Ebola outbreak on the vaccination of children in a rural district of Guinea.

Setting: All health centres in Macenta District, rural Guinea. Objective: To compare stock-outs of vaccines, vaccine stock cards and the administration of various childhood vaccines across the pre-Ebola, Ebola and post-Ebola virus disease periods. Design: This was an ecological study. Results: Similar levels of stock-outs were observed for all vaccines (bacille Calmette-Guérin [BCG], pentavalent, polio, measles, yellow fever) in the pre-Ebola and Ebola periods (respectively 2760 and 2706 facility days of stock-outs), with some variation by vaccine. Post-Ebola, there was a 65-fold reduction in stock-outs compared to pre-Ebola. Overall, 24 facility-months of vaccine stock card stock-outs were observed during the pre-Ebola period, which increased to 65 facility-months of stock-outs during the Ebola outbreak period; no such stock-out occurred in the post-Ebola period. Apart from yellow fever and measles, vaccine administration declined universally during the peak outbreak period (August-November 2014). Complete cessation of vaccine administration for BCG and a prominent low for polio (86% decrease) were observed in April 2014, corresponding to vaccine stock-outs. Post-Ebola, overall vaccine administration did not recover to pre-Ebola levels, with the highest gaps seen in polio and pentavalent vaccines, which had shortages of respectively 40% and 38%. Conclusion: These findings highlight the need to sustain vaccination activities in Guinea so that they remain resilient and responsive, irrespective of disease outbreaks.

Contexte: Tous les centres de santé de la Préfecture de Macenta, en Guinée rural.Objectif: Comparer la rupture en vaccins, en cartes de stock de vaccins et l'administration des différents vaccins d'enfance pendant les périodes pré-Ebola, Ebola et post-Ebola.Schéma: Une étude écologique.Résultats: Des niveaux similaires de rupture étaient observés pour tous les vaccins (bacille Calmette-Guérin [BCG], pentavalent, polio, rougeole, fièvre jaune) dans les périodes pré-Ebola et Ebola (respectivement 2760 et 2706 jours-structure de rupture), avec quelques variations par vaccin. Post-Ebola, il y avait 65 fois plus de réduction en rupture, comparé à la période pré-Ebola. Un total de 24 mois-structure de rupture en cartes de stock de vaccins était observé pendant la période pré-Ebola, qui a augmenté à 65 mois-structure de rupture pendant la période Ebola ; une telle rupture ne s'est pas produite dans la période post-Ebola. Excepté la fièvre jaune et la rougeole, l'administration de vaccin a diminué universellement pendant la période de pointe de l'épidémie (août­novembre 2014). L'arrêt complet de l'administration de vaccin pour le BCG et une baisse marquée pour la polio (diminution de 86%) étaient observés en avril 2014, correspondant à une rupture de vaccins. Post-Ebola, l'administration globale de vaccins n'a pas atteint les niveaux pré-Ebola, avec les plus grands écarts observés aux niveaux de la polio et du pentavalent (respectivement des baisses de 40% et 38%).Conclusion: Ces résultats soulignent le besoin de maintenir les activités de vaccination en Guinée afin qu'elles restent résilientes et réactives, indépendamment de l'épidémie d'une maladie.

Marco de referencia: Todos los centros de atención de salud del distrito de Macenta en una zona rural de Guinea.Objetivo: Comparar el desabastecimiento de vacunas, las tarjetas de existencias de vacunas y la administración de las diversas vacunas de la infancia durante diferentes períodos, en función de la epidemia de fiebre hemorrágica del Ébola, a saber: antes, durante el brote y después del mismo.Método: Un estudio ecológico.Resultados: Se observaron niveles equivalentes de desabastecimientos de todas las vacunas (BCG, pentavalente, antipoliomielítica, antisarampionosa y antiamarílica) antes de la epidemia del Ébola y durante la misma (2760 y 2706 días de desabastecimiento por establecimiento, respectivamente), con alguna variación en función de las vacunas. En el período posterior a la epidemia se presentó una tasa de desabastecimientos 65 veces menor, en comparación con el período anterior a la epidemia. En general, se observaron 24 meses-centro de desabastecimiento en las tarjetas de existencias vacunales durante el período pre-Ébola, que aumentaron a 65 meses-centro de desabastecimiento durante la epidemia; en el período posterior al brote no ocurrió este tipo de desabastecimiento. Con la excepción de la vacuna antiamarílica y la antisarampionosa, la administración de vacunas disminuyó globalmente durante el período de máxima actividad de la epidemia (de agosto a noviembre del 2014). Se observó una interrupción total de la administración de BCG y una tasa considerablemente baja de administración de vacuna antipoliomielítica (disminución de un 86%) en abril del 2014, que correspondió con el desabastecimiento de vacunas. Después de la epidemia del Ébola, la administración general de vacunas no recuperó el nivel anterior al brote y las mayores carencias se observaron con la vacuna antipoliomielítica y la pentavalente (40% y 38% de déficit, respectivamente).Conclusión: Los resultados del presente estudio destacan la necesidad de sostener las actividades de vacunación en Guinea, de manera que conserven su capacidad de recuperación y de respuesta, con independencia de los brotes epidémicos.

Molecular Identification of Malassezia Species in Patients with Malassezia folliculitis in Sfax, Tunisia.

AIM: Malassezia folliculitis is caused by the invasion of hair follicles by large numbers of Malassezia cells. Several Malassezia researches still use cultures, morphology and biochemical techniques. The aim of this study was to identify Malassezia species isolated from patients diagnosed with folliculitis, at the Parasitology and Mycology Laboratory of Sfax University Hospital, and to explore the genetic diversity of Malassezia by using PCR-RFLP and PCR-sequencing targeting the rDNA region of the Malassezia genome. PATIENTS AND METHODS: Specimens were taken from 27 patients with Malassezia folliculitis. For the molecular identification, PCR amplification of the 26S rDNAD1/D2 region was carried out using the Malup and Maldown primers and three restriction enzymes (BanI, MspI and HeaII) for RFLP analysis. The nucleotide sequences of each isolate were compared to those in the NCBI GenBank by using BLASTIN algorithm. RESULTS: Three species of Malassezia yeasts were identified among the 31 Malassezia strains isolated: M. globosa (83.9%), M. sympodialis (12. 9%) and M. furfur (3.2%). The sequence analysis of M. globosa showed six genotypes. CONCLUSION: There is a high genotypic variability of M. globosa colonizing patients with folliculitis.

Analysis of virulence factors and in vivo biofilm-forming capacity of Yarrowia lipolytica isolated from patients with fungemia.

Yarrowia lipolytica is ubiquitous in the environment, opportunistic, and might be considered as one of the causative agents of catheter-related candidemia. Our work aimed to study some virulence factors of Y. lipolytica such as hydrolases production and biofilm formation with comparison to the most frequent Candida specie in human disease. In sum, 58 clinical isolates of Y. lipolytica, 16 C. glabrata, and 12 C. albicans were collected from Intensive care unit (ICU). All were tested for enzymatic production and biofilm formation. All tested isolates of C. albicans and C. glabrata were able to degrade casein, and 98.2% of Y. lipolytica showed caseinase activity but no gelatinase activity was detected in all isolates. Y. lipolytica strains showed significantly lower (3.4%) in vitro phospholipase activity than C. albicans and C. glabrata (P < .05). No significant differences of the hemolytic activity were detected between the three species (P > .05). Concerning biofilm formation, and unlike the results obtained on polystyrene plate, the number of adhered and biofilm cultivable cells obtained by Y. lipolytica after 168 hours of catheter subcutaneous implantation is significantly greater and tends to be more compact and structured hyphal layer. Although C. albicans remains the most pathogenic yeast, development of selective ability of Y. lipolytica to adhere, to form a biofilm on catheter medical devices, and to produce phospholipase and hemolytic enzyme is of particular interest, and it is strongly recommended to be vigilant in the use of medical implanted medical devices, particularly in ICU.