Potential fungicidal and antiaflatoxigenic effects of cinnamon essential oils on Aspergillus flavus inhabiting the stored wheat grains.

Wheat is one of the essential crops for the human and animal nutrition, however, contamination with aflatoxigenic fungi, due to the improper storage conditions and high humidity, was the main global threats. So, preventing the growth of aflatoxigenic fungi in stored wheat grains, by using different essential oils was the main objective of this work. Aspergillus flavus EFBL-MU12 PP087400, EFBL-MU23 PP087401 and EFBL-MU36 PP087403 isolates were the most potent aflatoxins producers inhabiting wheat grains. The effect of storage conditions of wheat grains "humidity, temperature, incubation period, and pH" on growth of A. flavus, was assessed by the response surface methodology using Plackett-Burman design and FCCD. The highest yield of aflatoxins EFBL-MU12 B1 and B2 by A. flavus grown on wheat grains were 145.3 and 7.6 µg/kg, respectively, at incubation temperature 35°C, 16% moisture contents, initial pH 5.0, and incubated for 14 days. The tested oils had a powerful antifungal activity for the growth and aflatoxins production by A. flavus in a concentration-dependent manner. Among these oils, cinnamon oil had the highest fungicidal activity for A. flavus at 0.125%, with about 85-90 % reduction to the aflatoxins B1 and B2, conidial pigmentation and chitin contents on wheat grains. From the SEM analysis, cinnamon oils had the most deleterious effect on A. flavus with morphological aberrations to the conidial heads, vegetative mycelia, alteration in conidiophores identity, hyphae shrank, and winding. To emphasize the effect of the essential oils on the aflatoxins producing potency of A. flavus, the molecular expression of the aflatoxins biosynthetic genes was estimated by RT-qPCR. The molecular expression of nor-1, afLR, pKsA and afLJ genes was suppressed by 94-96%, due to cinnamon oil at 0.062% compared to the control. Conclusively, from the results, cinnamon oils followed by the peppermint oils displayed the most fungicidal activity for the growth and aflatoxins production by A. flavus grown on wheat grains.

Application of artificial neural networks for enhancing Aspergillus flavipes lipase synthesis for green biodiesel production.

Biodiesel is a sustainable, and renewable alternative to fossil fuels that can be produced from various biological sources with the aid of lipases. This study developed a simple and novel fungal system for lipase biosynthesis to be used for catalyzing the oily residuals into biodiesel, employing the artificial neural network (ANN), and semi-solid-state fermentation (SSSF). Nigella sativa was selected among agro-industrial oily residuals as a substrate for lipase biosynthesis by Aspergillus flavipes MH47297. The effect of cultural humidity (X1), the surfactant; Brij 35 (X2), and inoculum density (X3) on lipase biosynthesis were researched based on the matrix of Box-Behnken design (BBD). The ANN together with a new fungal candidate and SSSF were then applied for the first time to model the biosynthesis process of lipase. The optimum predicted cultural conditions varied according to the model. The optimum predicted conditions were estimated separately by BBD (X1 = 5.8 ml water/g, X2 = 46.6 µl/g, and X3 = 62156610 spore/g) and ANN (X1 = 5.4 ml water/g, X2 = 54.2 µl/g, and X3 = 100000000 spore/g) models. Based on the modeling process, the response of lipase was calculated to be 214.95 (BBD) and 217.72 U (ANN), which revealed high consistency with the experimental lipase yield (209.13 ± 3.27 U for BBD, and 218 ± 2.01 U for ANN). Despite both models showing high accuracy, ANN was more accurate and surpassed the BBD model. Gas chromatography analysis showed that lipase successfully converted corn oil to biodiesel (29.5 mg/l).

Production, bioprocessing and antiproliferative activity of camptothecin from Aspergillus terreus, endophyte of Cinnamomum camphora: restoring their biosynthesis by indigenous microbiome of C. camphora.

Fungal producing potency of camptothecin (CPT) raise the hope for their usage to be a platform for industrial production of CPT, nevertheless, attenuation of their productivity of CPT with the subculturing and preservation is the challenge. So, screening for novel endophytic fungal isolates with a reliable CPT-biosynthetic stability was the objective. Among the isolated endophytic fungi from the tested medicinal plants, Aspergillus terreus OQ642314.1, endophyte of Cinnamomum camphora, exhibits the highest yield of CPT (89.4 µg/l). From the NMR, FT-IR and LC-MS/MS analyses, the extracted CPT from A. terreus gave the same structure and molecular mass fragmentation pattern of authentic CPT (349 m/z). The putative CPT had a significant activity against MCF7 (0.27 µM) and HEPG-2 (0.8 µM), with a strong affinity to inhibits the human Topoisomerase 1 activity (IC50 0.362 µg/ml) as revealed from the Gel-based DNA relaxation assay. The purified CPT displayed a strong antimicrobial activity for various bacterial (E. coli and B. cereus) and fungal (A. flavus and A. parasiticus) isolates, ensuring the unique tertiary, and stereo-structure of A. terreus for penetrating the microbial cell walls and targeting the topoisomerase I. The higher dual activity of the purified CPT as antimicrobial and antitumor, emphasize their therapeutic efficiency, especially with growth of the opportunistic microorganisms due to the suppression of human immune system with the CPT uses in vivo. The putative CPT had an obvious activity against the tumor cell (MCF7) metastasis, and migration as revealed from the wound healing assay. The overall yield of A. terreus CPT was maximized with the Blackett-Burman design by twofolds increment (164.8 µg/l). The CPT yield by A. terreus was successively diminished with the multiple fungal subculturing, otherwise, the CPT productivity of A. terreus was restored, and increased over the zero culture upon coculturing with C. camphora microbiome (1.5% w/v), ensuring the restoring of CPT biosynthetic potency of A. terreus by the plant microbiome-derived chemical signals "microbial communication". This is the first report exploring the feasibility of A. terreus "endophyte of C. camphora" to be a preliminary platform for commercial production of CPT with a reliable sustainability upon uses of indigenous C. camphora microbiome.

Furlow Palatoplasty and Tonsillectomy for Treating Patients With Submucous Cleft Palate and Tonsillar Hypertrophy: A One-Stage Procedure.

BACKGROUND: Children with cleft palate are more liable to have obstructive sleep apnea than children with normal palate due to narrow airways. Tonsillar hypertrophy is a common cause of pediatric obstructive sleep apnea; hence, it is not surprising to be encountered during cleft palate repair. The aim of this study was to evaluate the feasibility of tonsillectomy and Furlow palatoplasty performed as a 1-stage operation in patients presenting with submucous cleft palate (SMCP) and tonsillar hypertrophy. MATERIALS AND METHODS: Eleven pediatric patients with SMCP and hypertrophied tonsils were included in this case series study. Furlow palatoplasty and tonsillectomy were performed for the patients in 1 sitting. The evaluation of velopharyngeal function was done preoperatively and postoperatively via auditory-perceptual-assessment, nasometry, and flexible nasopharyngoscopy. In addition, the Epworth sleepiness scale for children/adolescents was administered to the parents to assess daytime sleepiness of their children. RESULTS: The speech improved postoperatively. Auditory-perceptual-assessment showed significant reductions in hypernasal speech, nasal air escape, and weak pressure consonants. In addition, nasometry revealed significantly decreased nasalance scores for nasal and oral sentences. A postoperative increased velar movement was observed with a significant improvement in velopharyngeal closure. The preoperative Epworth sleepiness scale for children/adolescents assessment revealed excessive daytime sleepiness in 8 patients, with significant improvement of scores postoperatively. CONCLUSIONS: Removal of hypertrophied tonsils during the repair of SMCP with Furlow palatoplasty did not negatively affect speech outcome or velar movement postoperatively. It is logical to perform both procedures simultaneously in 1 sitting to avoid postoperative sleep-related breathing disorder, which may necessitate a second stage operation.

The Healing Capability of Clove Flower Extract (CFE) in Streptozotocin-Induced (STZ-Induced) Diabetic Rat Wounds Infected with Multidrug Resistant Bacteria.

Treatment of diabetic foot ulcer (DFU) is of great challenge as it is shown to be infected by multidrug resistant bacteria (MDR bacteria). Sixty four bacterial isolates were isolated from DFU cases; antibiotic susceptibility tests were carried out for all of them. One bacterial isolate (number 11) was shown to resist the action of 8 out of 12 antibiotics used and was identified by both a Vitek-2 system and 16S rRNA fingerprints as belonging to Proteus mirabilis, and was designated Proteus mirabilis LC587231 (P. mirabilis). Clove flower extract (CFE) inhibited distinctively the P. mirabilis bacterium obtained. GC-MS spectroscopy showed that this CFE contained nine bioactive compounds. The effect of CFE on wound healing of Type 1 diabetic albino rats (Rattus norvegicus) was studied. The results indicated that topical application of CFE hydrogel improved wound size, wound index, mRNA expression of the wound healing markers (Coli1, MMP9, Fibronectin, PCNA, and TGFß), growth factor signaling pathways (PPAR-α, PGC1-α, GLP-1, GLPr-1, EGF-ß, EGF-ßr, VEGF-ß, and FGF-ß), inflammatory cytokine expression (IL8, TNFα, NFKß, IL1ß, and MCP1), as well as anti-inflammatory cytokines (IL4 & IL10), pro-apoptotic markers (FAS, FAS-L, BAX, BAX/BCL-2, Caspase-3, P53, P38), as well as an antiapoptotic one (BCL2). Furthermore, it improved the wound oxidative state and reduced the wound microbial load, as the cefepime therapy improved the wound healing parameters. Based on the previous notions, it could be concluded that CFE represents a valid antibiotics alternative for DFU therapy since it improves diabetic wound healing and exerts antibacterial activity either in vitro or in vivo.

Nasopharyngeal Polyp in a Patient With Submucous Cleft Palate.

Encountering a nasopharyngeal polyp in a patient with submucous cleft palate (SMCP) is a difficult problem, as the lesion could support the weak palate. Removal of this lesion may unmask the SMCP with consequent worsening of speech nasality. Nasal septal polyp protruding to the nasopharynx in a patient with SMCP has not been reported before in the literature. This report describes a septal polyp arising from the posterior border of the nasal septum and protruding in the nasopharynx in a 16-year-old girl with submucous cleft palate. The polyp appeared to support the weak palate, and they acted as a ball and socket during speech articulation. Removal of this polyp may result in velopharyngeal insufficiency. Trans-nasal endoscopic removal of the polyp with obturation of the velopharyngeal port with a superiorly-based pharyngeal flap was performed in the same sitting. Pre- and postoperative speech evaluation using auditory perceptual assessment and nasometry revealed no worsening of nasality, also the patient reported improvement of her nasal breathing. We concluded that, the presence of a nasopharyngeal polyp in a patient with SMCP may compensate the speech problem. Removal of the polyp and treatment of SMCP by a pharyngeal flap in one-sitting is an effective procedure without adverse effect on patient's speech.

Detection and Control of Biofilm Formation by Staphylococcus aureus from Febrile Neutropenic Patient.

INTRODUCTION: Febrile neutropenia (FN) is the evolution of fever in a patient with neutropenia over 38.0°C. Neutropenia is diagnosed when absolute neutrophil count (ANC) <1500 cells/µL. FN represents a common complication of cancer treatment. Hence, it is featured to be a major cause of morbidity and mortality in cancer patients. Staphylococcus aureus is one of the most important microorganisms isolated from the blood of febrile neutropenic patients. Infections caused by S. aureus range from mild to life-threatening diseases. Biofilm production by S. aureus is one of the most significant virulence factors of the bacterium as it prevents the penetration of antibiotics. Recently, it has been shown that S. aureus carries the ica operon responsible for biofilm production. The aim of the work is to determine a genotypic characterization that includes not only the detection of icaA and icaD genes in S. aureus but also the determination of their relation to clinical and microbiological features. Empiric antibacterial treatment was recommended for cancer patients receiving chemotherapy. MATERIALS AND METHODS: The relation between the presence of icaA and icaD and biofilm production was determined in a collection of 66 S. aureus samples from febrile neutropenic patients. Biofilm-forming ability was tested on Congo Red agar plates. Also, the effect of the most sensitive antibiotics on the bacterial cells was determined by an electron microscope. RESULTS: Of the bacterial samples, 48 were biofilm-productive and 18 were non-biofilm productive. For the biofilm productive bacteria, 37.5% were positive for icaA, 22.9% were positive for icaD and 10.4% were positive for both. Linezolid was the most effective antibiotic and it is highly recommended for the treatment of febrile neutropenia caused by biofilm-productive S. aureus. Severe changes were found on the bacterial cell after being treated with Linezolid. The icaA and icaD genes were present in only 50% of biofilm-productive bacteria. CONCLUSION: The ica operon is present in only 50% of biofilm-productive S. aureus and Linezolid is the best antibiotic against these bacteria.

Phytochemical, antimicrobial and antiquorum-sensing studies of pulicaria undulata L.: a revision on the structure of 1ß,2α,3ß,19α,23-pentahydroxy-urs-12-en-28-oic acid.

Phytochemical study of the aerial part of Pulicaria undulata L. led to the isolation of nine compounds. The structure of 1ß,2α,3ß,19α,23-pentahydroxy-urs-12-en-28-oic acid (4) was revised and confirmation of the stereochemical configuration of the hydroxyl groups was established using NOESY and selective decoupling experiments. The other compounds were identified as 1,2-dehydro-1,10α-dihydropseudoivalin (1), axillarin (2), grandifloric acid-15-ß-glucoside (3), myrianthic acid (5), caffeic acid (6), quercetin (7), paniculoside IV (8) and caffeic anhydride (9). The structures were characterized by 1 D, 2 D NMR spectroscopy and confirmed with HRMS. Antimicrobial and antiquorum-sensing activities of the different extracts and isolated compounds of the plant were investigated. Generally, the phenolic rather than the terpenoidal compounds exhibited remarkable antimicrobial and antiquorum-sensing activity.[Formula: see text].

Potential of fungal metabolites as a biocontrol agent against cotton aphid, Aphis gossypii Glover and the possible mechanisms of action.

The present study investigated the insecticidal activity of the different organic extracts from the entomopathogenic fungi, Cladosporium cladosporioides, Metarhizium anisopliae, Purpureocillium lilacinum, and Trichoderma longibrachiatum towards cotton aphid, Aphis gossypii. The methanol extracts from the mycelia and spores of C. cladosporioides and P. lilacinum exhibited the highest insecticidal activity against A. gossypii compared with other extracts, which LC50 values were recorded to be 57.60 and 94.18 ppm, respectively. The major constituents identified in both methanol extracts by GC-MS analysis were linoleic acid and palmitic acid. The methanol extracts of C. cladosporioides and P. lilacinum caused a voluminous increase in the total carbohydrates content of A. gossypii adults, while the total protein content was significantly decreased by both extracts. The activity of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were significantly reduced by methanol extracts. The P. lilacinum extract caused a considerable reduction in the activity of glutathione-S-transferase (GST), α- and ß-esterase by 28.9, 27.9 and 23.4%, respectively. Both extracts induced a significant increase in phenoloxidase and chitinase activity of A. gossypii adults. These results suggest that C. cladosporioides and P. lilacinum methanol extracts could be used as a promising approach for the management of A. gossypii in many economically crops.

Palatal eversion for the treatment of combined nasopharyngeal stenosis and tonsillar pillars adhesion.

OBJECTIVE: Rarely the tonsillar pillars and the soft palate became adherent to the posterior nasopharyngeal wall by strong fibrous tissue due to excessive dissection and cauterization during surgery leading to nasopharyngeal stenosis. Therefore, many treatment modalities are being tried to cure this problem. The aim of this study is to explore our results of modifying the basic technique to accommodate those patients with combined nasopharyngeal stenosis and tonsillar pillars adhesions in one stage. STUDY DESIGN: Case series. METHODS: This study was conducted on 10 patients with combined nasopharyngeal stenosis and tonsillar pillars adhesions after adenotonsillectomy. They were subjected to treatment by palatal eversion through dividing the soft palate in the midline to separate each pillar from the pharyngeal wall in continuation with each half of soft palate and removal of the fibrous tissue causing stenosis. This was followed by eversion and fixation of the two palatal divisions on either side to allow complete epithelialization of the stenotic area. Postoperative follow-up was done for one year by the flexible nasopharyngoscopy, perceptual speech analysis, and polysomnography. RESULTS: The flexible nasopharyngosopic examination of the 10 patients at the end of post-operative period revealed a freely mobile soft palate with no nasopharyngeal stenosis or palatal fistula. Velopharyngeal function and speech assessment by perceptual speech analysis was normal in all 10 cases. No obstructive episodes were recorded in polysomnograms. CONCLUSIONS: Palatal eversion is a promising technique in the treatment of post-adenotonsillectomy of combined nasopharyngeal stenosis and tonsillar pillars adhesion. It is recommended to be used on a wider scale of patients and other indications as nasopharyngeal stenosis following uvulopalatoplasty and post nasopharyngeal radiotherapy. The level of evidence: 4 (case series).

Fluorescence enhancement monitoring of pyrromethene laser dyes by metallic Ag nanoparticles.

Fluorescence enhancement monitoring of pyrromethene laser dyes using their complexation with Ag nanoparticles (Ag NPs) was studied. The size of the prepared Ag NPs was determined by transmission electron spectroscopy and UV/Vis absorption spectroscopy. Mie theory was also used to confirm the size of NPs theoretically. The effect of different nanoparticle concentrations on the optical properties of 1 × 10(-4) M PM dyes shows that 40%of Ag NPs concentration (40%C Ag NPs) in complex is the optimum concentration. Also, the effects of different concentrations of PM dyes in a complex was measured. Emission enhancement factors were calculated for all samples. Fluorescence enhancement efficiencies depended on the input pumping energy of a Nd-YAG laser (wavelength 532 nm and 8 ns pulse duration) were reported and showed the lowest energy (28 and 32 mJ) in the case of PM567 and PM597, respectively.

Trichoderma harzianum: a biocontrol agent against Bipolaris oryzae.

Rice brown spot, caused by Bipolaris oryzae, can be a serious disease causing a considerable yield loss. Trichoderma harzianum is an effective biocontrol agent for a number of plant fungal diseases. Thus, this research was carried out to investigate the mechanisms of action by which T. harzianum antagonizes Bipolaris oryzae in vitro, and the efficacy of spray application of a spore suspension of T. harzianum for control of rice brown spot disease under field conditions. In vitro, the antagonistic behavior of T. harzianum resulted in the overgrowth of B. oryzae by T. harzianum, while the antifungal metabolites of T. harzianum completely prevented the linear growth of B. oryzae. Light and scanning electron microscope (SEM) observations showed no evidence that mycoparasitism contributed to the aggressive nature of the tested isolate of T. harzianum against B. oryzae. Under field conditions, spraying of a spore suspension of T. harzianum at 10(8)spore ml(-1) significantly reduced the disease severity (DS) and disease incidence (DI) on the plant leaves, and also significantly increased the grain yield, total grain carbohydrate, and protein, and led to a significant increase in the total photosynthetic pigments (chlorophyll a and b and carotenoids) in rice leaves.