Endophytic fungal community of Rosa damascena Mill. as a promising source of indigenous biostimulants: Elucidating its spatial distribution, chemical diversity, and ecological functions.

The role of endophytes in maintaining healthy plant ecosystems and holding promise for agriculture and food security is deeply appreciated. In the current study, we determine the community structure, spatial distribution, chemical diversity, and ecological functions of fungal endophytes of Rosa damascena growing in the North-Western Himalayas. Culture-dependent methods revealed that R. damascena supported a rich endophyte diversity comprising 32 genera and 68 OTUs. The diversity was governed by climate, altitude, and tissue type. Species of Aspergillus, Cladosporium, Penicillium, and Diaporthe were the core endophytes of the host plant consisting of 48.8% of the endophytes collectively. The predominant pathogen of the host was Alternaria spp., especially A. alternata. GC-MS analyses affirmed the production of diverse arrays of volatile organic compounds (VOC) by individual endophytes. Among the primary rose oil components, Diaporthe melonis RDE257, and Periconia verrucosa RDE85 produced phenyl ethyl alcohol (PEA) and benzyl alcohol (BA). The endophytes displayed varied levels of plant growth-promoting, colonization, and anti-pathogenic traits. Between the selected endophytes, P. verrucosa and D. melonis significantly potentiated plant growth and the flavonoids and chlorophyll content in the host. The potential of these two endophytes and their metabolites PEA and BA was confirmed on Nicotiana tabacum. The treatments of the metabolites and individual endophytes enhanced the growth parameters in the model plant significantly. The results imply that P. verrucosa and D. melonis are potential plant growth enhancers and their activity may be partially due to the production of PEA and BA. Thus, R. damascena harbors diverse endophytes with potential applications in disease suppression and host growth promotion. Further investigations at the molecular level are warranted to develop green endophytic agents for sustainable cultivation of R. damascena and biocontrol of leaf spot disease.

Vasorelaxant property of 2-phenyl ethyl alcohol isolated from the spent floral distillate of damask rose (Rosa damascena Mill.) and its possible mechanism.

ETHNOBOTANICAL RELEVANCE: Rosa damascena Mill. (Rosaceae), commonly known as damask rose, is an ancient medicinal and perfumery plant used in Traditional Unani Medicine due to various therapeutic effects, including cardiovascular benefits. AIM OF THE STUDY: This study aimed to evaluate the vasorelaxant effect of the 2-phenyl ethyl alcohol (PEA) isolated from the spent flowers of R. damascena which remain after the extraction of essential oil. MATERIALS AND METHODS: The freshly collected flowers of R. damascena were hydro-distilled in a Clevenger's type apparatus to extract the rose essential oil (REO). After removing the REO, the spent-flower hydro-distillate was collected and extracted with organic solvents to yield a spent-flower hydro-distillate extract (SFHE), which was further purified by column chromatography. The SFHE and its isolate were characterized by gas chromatography (GC-FID), gas chromatography-mass spectrometry (GC-MS), and nuclear magnetic resonance (NMR) techniques. The PEA, isolated from SFHE, was evaluated for vasorelaxation response in conduit blood vessels like rat aorta and resistant vessels like mesenteric artery. The preliminary screening of PEA was done in aortic preparation pre-constricted with phenylephrine/U46619. Further, a concentration-dependent relaxation response to PEA has been elicited in both endothelium-intact and endothelium-denuded arterial rings, and the mode of action was explored. RESULTS: The SFHE revealed the presence of PEA as the main constituent (89.36%), which was further purified by column chromatography to a purity of 95.0%. The PEA exhibited potent vasorelaxation response both in conduit vessels like the rat aorta and resistance vessels like the mesenteric artery. The relaxation response is mediated without any involvement of vascular endothelium. Further, TEA sensitive BKCa channel was found to be the major target for PEA-induced relaxation response in these blood vessels. CONCLUSIONS: The spent flowers of R. damascena, which remain after the extraction of REO, could be used to extract PEA. The PEA possessed marked vasorelaxation properties in both aorta and mesenteric artery and showed promise for development into an herbal product against hypertension.

Bacterial diversity of a floating vegetation (Phumdi) of Loktak Lake and its extracellular enzymes and bacterial antagonistic property.

134 bacterial strains were isolated from phumdis of Loktak Lake. Through 16S rRNA sequencing, Bacillus sp. (23, 17.1%), Staphylococcus sp. (14, 10.4%), Pseudomonas sp. (11, 8.2%) and Acinetobacter sp. (8, 5.9%) were identified as the predominant bacterial taxa of Loktak Lake. B. pumulis (12, 8.9%), S. arlettae (4, 2.9%), P. knackmussii (6, 4.4%) are the leading species of Bacillus, Staphylococcus and Pseudomonas, respectively. Similarly, A. seifertii (2, 1.4%) and A. calcoaceticus (2, 1.4%) are the common species of Acinetobacter. 75 (55.9%) bacterial strains showed the ability to hydrolyze one or more extracellular enzymes tested. Among the extracellular enzymes produced by the bacterial isolates, the presence of elastase activity cannot be underestimated, since the enzyme is involved in the process of bacterial lung infection. Phosphate solubilizing activity could be seen in 11.1% of the bacterial isolates. 27 (20.1%) of the strains shown to have antagonistic activity against one or more tested pathogens. An isolate, MRC 52 showed antagonistic activity against eleven different pathogens including carbapenem resistant E. coli which was further subjected to extraction and identification of the biomolecule exerting the antimicrobial property. Based on GC-MS analysis, the bioactive compound was identified as phenyl ethyl alcohol.

Unravelling the Novel Effects of Three Volatile Compounds in Preventing Fibril Formation of Disease Related Tau and α-Synuclein Proteins- Towards Identifying Candidate Aromatic Substances for Treating Neurodegenerative Diseases.

Background: The aggregation of tau and α-synuclein into fibrillary assemblies in nerve cells is the molecular hallmark of Alzheimer's and Parkinson's diseases, respectively. In our previous studies, we investigated the anti-amyloidogenic effects of three different aroma-producing (volatile) compounds including cinnamaldehyde, phenyl ethyl alcohol, and TEMED on the fibrillation process of HEWL, as a model protein. Our previous results showed that while TEMED was able to completely stop the process of fibril formation, cinnamaldehyde and phenyl ethyl alcohol gave rise to oligomeric/protofibrillar forms and were involved in the entrapment of intermediate species of HEWL. In this study, we investigated the anti-amyloidogenic effect of the same three volatile compounds on recombinantly produced tau and α-synuclein proteins. Methods: The thioflavin T fluorescence assay, circular dichroism, SDS-PAGE/native-PAGE, dynamic light scattering, and atomic force microscopy were used, where necessary, to further our understanding of the inhibitory effects of the three volatile compounds on the fibril formation of tau and α-synuclein proteins and allow for a comparison with previous data obtained for HEWL. Results: Our results revealed that contrary to the results obtained for HEWL (a globular protein), the volatile compound TEMED was no longer able to prevent fibril formation in either of the natively unstructured tau or α-synuclein proteins, and instead, cinnamaldehye and phenyl ethyl alcohol, in particular, had the role of preventing fibril formation of tau or α-synuclein. Conclusion: The results of this study further emphasized the exclusion of HEWL as a model protein for fibrillation studies and highlighted the importance of studying brain-related proteins such as tau or α-synuclein and the need to assess the effects of volatile compounds such as cinnamaldehye and phenyl ethyl alcohol as potential substances in the treatment of neurodegenerative diseases.

Quantification of the Geranium Essential Oil, Palmarosa Essential Oil and Phenylethyl Alcohol in Rosa damascena Essential Oil Using ATR-FTIR Spectroscopy Combined with Chemometrics.

Rosa damascena essential oil is an essential oil that has the greatest industrial importance due to its unique quality properties. The study used ATR-FTIR (attenuated total reflectance-Fourier transform infrared) spectroscopy coupled with chemometrics of PLSR (partial least squares regression) and PCR (principal component regression) for quantification of probable adulterants of geranium essential oil (GEO), palmarosa essential oil (PEO) and phenyl ethyl alcohol (PEOH). Hierarchical cluster analysis was performed to observe the classification pattern of Rosa damascena essential oil, spiked samples and adulterants. Rosa damascena essential oil was spiked with each adulterant at concentrations of 0-100% (v/v). Excellent R2 (regression coefficient) values (≥0.96) were obtained in all PLSR and PCR cross-validation models. The SECV (standard error of cross-validation) values ranged between 0.43 and 4.15. The lowest SECV and bias values were observed in the PLSR and PCR models, which were built by using the raw FTIR spectra of all samples. Hierarchical cluster analysis through Ward's algorithm and Euclidian distance had high potential to observe the classification pattern of all adulterated and authentic samples. In conclusion, the combination of ATR-FTIR spectroscopy with multivariate analysis can be used for rapid, cost-effective, easy, reliable and high-throughput detection of GEO, PEO and PEOH in Rosa damascena essential oil.

Subcritical Extracts from Major Species of Oil-Bearing Roses-A Comparative Chemical Profiling.

A comprehensive chemical profiling of 1,1,1,2-tetrafluoroethane (freon R134a) subcritical extracts from the main genotypes of oil-bearing roses, was performed by gas chromatography-mass spectrometry (GC/MS) and gas chromatography with flame ionization detection (GC-FID) in order to reveal the differences in their chemical composition. One hundred and three individual compounds were identified using GC/MS and their quantitative content was determined using GC-FID, representing 89.8, 92.5, 89.7 and 93.7% of the total content of Rosa gallica L., Rosa damascena Mill., Rosa alba L. and Rosa centifolia L. extracts, respectively. The compounds found in the extracts are representatives of the following main chemical classes: mono-, sesqui- and triterpenoids, phenylethanoids and phenylpropanoids and aliphatic hydrocarbons. Fatty acids, esters and waxes were found, as well. The study revealed that 2-phenylethanol is the most abundant component, ranging 9.0-60.9% followed by nonadecane and nonadecene with 5.1-18.0% geraniol (2.9-14.4%), heneicosane (3.1-11.8%), tricosane (0.1-8.6%), nerol (1.3-6.1%) and citronellol (1.7-5.3%). The extracts demonstrate a specific chemical profile, depending on the botanical species-phenylethanoids and phenyl propanoids are the main group for R. damascena, aliphatic hydrocarbons for R. alba and R. centifolia, while both are found in almost equal amounts in R. gallica. The terpenoid compounds show relatively broad variations: monoterpenes-11.9-25.5% with maximum in R. centifolia; sesquiterpenes-0.6-7.0% with maximum in R. gallica and triterpenes-0.4-3.7% with maximum in R. gallica extract.

A study of 30 odors panel smell identification test, smell detection threshold and University of Pennsylvania Smell Identification Test (UPSIT) in Thailand.

OBJECTIVES: This study aims to evaluate the application of the University of Pennsylvania Smell Identification Test (UPSIT) in Thailand, as well as to conduct an odor detection threshold test using phenyl ethyl alcohol and a 30-odor smell identification test. METHODS: This study was conducted from May 2019 to March 2020. We included healthy volunteers without any olfactory complaints. All participants underwent the UPSIT, an evaluation of odor detection threshold, and a 30-odor smell identification test. RESULTS: One hundred fifty participants were included in our study. The overall mean score on the UPSIT was 26.04 ± 6.59 points and ranged from 9 to 39.7. For participants aged under 60 years, the mean UPSIT score was 29.08 ± 4.67 points, while the mean score of those 60 years of age and over was 19.20 ± 4.97 points (mean difference of 9.88 points), a difference that was statistically significant (95% CI 8.23 to 11.53, p<0.0001). The mean lowest log value on the odor detection threshold test was -7.12 ± 1.64. This was -7.53 ± 1.05 in participants under 60 years of age and -6.20 ± 2.27 in those 60 years of age and over (mean difference of 1.33; 95% CI 0.80 to 1.86, p<0.0001). Items in the 30-odor smell identification test with correct response rates greater than 70% included fish sauce, banana, coffee, patchouli water, coconut, lemongrass, orange, ammonia, vinegar, tea leaf, Thai perfume, jasmine, pandan, curry, lime, durian, cola, corn, pineapple, strawberry, and grape. CONCLUSIONS: This study identified the odor detection threshold, UPSIT scores, and suitable odors to use in smell identification in a Thai population.

The role of Cinnamaldehyde and Phenyl ethyl alcohol as two types of precipitants affecting protein hydration levels.

Release of water is the main force which drives proteins towards crystallisation (giving rise to protein crystals for crystallography) and aggregation (main cause of neurodegenerative diseases), and as such it is possible to make changes in the crystallisation/aggregation process by using compounds which are able to reduce the amount of water molecules around proteins. Cinnamaldehyde and Phenyl ethyl alcohol are the active constituents of cinnamon and rose flower, respectively. Traditional Iranian Medicine (TIM) suggests the use of cinnamon and rose flower for the reduction of excess coldness and wetness from the brain of patients suffering from Dementia. Using crystallisation as a model system and X-ray crystallography, we tested whether Cinnamaldehyde or Phenyl ethyl alcohol can mimic the role of precipitants resulting in the formation of crystals of HEWL (as a model protein) by releasing water from the surrounding protein environment. Results have revealed that both Cinnamaldehyde and Phenyl ethyl alcohol, in particular, were capable to adequately act as 'precipitants' but in the presence of NaCl (as a salt), resulting in better crystals of HEWL by changing the amount of charge and/or making water molecules unavailable in the symmetry related position, in line with the role suggested for these compounds by TIM.

The effect of olfactory training on odor identification in patients with traumatic anosmia.

BACKGROUND: This study investigates the effect of olfactory training on odor identification in patients with traumatic anosmia. METHODS: Patients with a clear history of loss of smell after head injury, and whose phenyl ethyl alcohol (PEA) odor detection thresholds remained at -1 after steroid and zinc treatment, were included in this study between July 2016 and July 2018. They were randomly divided into 2 groups, with patients in the 4-odorant group given 4 bottles of PEA, lemon, eucalyptus, and clove oils and those in the PEA group given a bottle of PEA for 6-month olfactory training. After 3-month and 6-month training, the olfactory function was evaluated by both the PEA threshold test and the traditional Chinese version of the University of Pennsylvania Smell Identification Test (UPSIT-TC). Magnetic resonance imaging was performed to measure the volume of olfactory bulbs after training. RESULTS: There were 45 patients completing 4-odorant training and another 45 completing PEA training. The birhinal PEA threshold decreased significantly in both groups after 6-month training, but the decrease was not significantly different between the 2 groups. The UPSIT-TC score increased significantly in the PEA group but not in the 4-odorant group. The volume of olfactory bulbs was not significantly different between these 2 groups. CONCLUSION: Our results show that olfactory training can slightly improve odor threshold levels in patients with traumatic anosmia, but did not improve the odor identification ability. Nevertheless, clinical improvement or benefit in quality of life from olfactory training needs further investigation.

FMRI correlates of olfactory processing in typically-developing school-aged children.

Human olfactory processing is understudied relative to other sensory modalities, despite its links to neurodevelopmental and neurodegenerative disorders. To address this limitation, we developed a fast, robust fMRI odor paradigm that is appropriate for all ages and levels of cognitive functioning. To test this approach, thirty-four typically developing children aged 7-12 underwent fMRI during brief, repeated exposure to phenylethyl alcohol, a flower-scented odor. Prior to fMRI scanning, olfactory testing (odor detection and identification) was conducted. During fMRI stimulus presentation, odorant release was synchronized to each participant's inspiratory phase to ensure participants were inhaling during the odorant exposure. Between group differences and correlations between activation and odor detection threshold scores were tested using the FMRIB Software Library. Results demonstrated that our 2-min paradigm significantly activated primary and secondary olfactory regions. In addition, a significant relationship between odor detection threshold and higher activation in the right amygdala and lower activation in the left frontal, insular, occipital, and cerebellar regions was observed, suggesting that this approach is sensitive to individual differences in olfactory processing. These findings demonstrate the feasibility of studying olfactory function in children using brain imaging techniques.

Favorite odor induces negative dream emotion during rapid eye movement sleep.

OBJECTIVE: As connections between nightmares and various psychiatric disorders have been reported, the clinical significance of studying dream emotionality is now growing in importance. Because the olfactory bulb connects directly to the amygdala odor presentation may be a crucial tool to study dream emotions. Previous studies have demonstrated that presentation of positive/negative odors during rapid eye movement (REM) sleep affects various aspects of dreaming. Although olfactory perception can be influenced by personal experiences, the role of individual preferences in the effects of olfactory stimuli on dreaming has not yet been clarified. The purpose of the current study was to clarify the effects of odor on dreaming during REM sleep, taking individual preferences into account. METHODS: Phenyl ethyl alcohol (rose-like smell) airflow was presented as an experimental stimulus, and odorless airflow was presented as the control. Participants who like (n = 7) and dislike (n = 8) the odor of phenyl ethyl alcohol were presented air with and without the odor of phenyl ethyl alcohol, respectively, during REM sleep and then awakened to report and rate their dream contents. Thereafter, the transcribed dream reports were rated by independent raters. RESULTS AND CONCLUSIONS: Participants who liked the odor of phenyl ethyl alcohol reported more emotionally negative dreams when they were presented with phenyl ethyl alcohol airflow than that with odorless airflow. In other words, the participant's favorite odor was associated with emotionally negative dreams. These findings could be attributed to the nature of odor perception and the characteristics of brain activities during REM sleep.

Establishment of olfactory diagnosis for the traditional Chinese version of the University of Pennsylvania Smell Identification Test.

BACKGROUND: This study attempted to establish an olfactory diagnosis for the traditional Chinese version of the University of Pennsylvania Smell Identification Test (UPSIT-TC). METHODS: Volunteers were enrolled to receive a phenyl ethyl alcohol (PEA) threshold test. Their olfactory function was divided into normal (threshold ≤ -6), mildly impaired (-6 < threshold ≤ -4), moderately impaired (-4 < threshold ≤ -2), and poor odor detection (threshold > -2). Subjects were also grouped by age: 20 to 29 years; 30 to 39 years; 40 to 49 years; 50 to 59 years; 60 to 69 years; and 70 to 79 years. They then received the UPSIT-TC to examine their smell identification function. The cutoff scores of UPSIT-TC among the different groups of olfactory function were determined by the receiver operating characteristic curve. RESULTS: A total of 1440 subjects were enrolled. The score of UPSIT-TC significantly decreased in both men and women after the age of 60 years. The cutoff scores were 29.5 between the normal and mildly impaired odor detection groups, 26.5 between the mildly and moderately impaired odor detection groups, and 16.5 between the moderately impaired and poor odor detection groups for males aged 20 to 59 years; the cutoff scores for these detection groups for males aged 60 to 79 years were 23.5. 20.5, and 13.5, respectively, and the cutoff scores for these detection groups for females aged 20 to 59 years were 30.5, 27.5, and 16.5, respectively. However, we did not find adequate cutoff scores for elderly females. CONCLUSION: This study established an olfactory diagnosis of UPSIT-TC for adult subjects. However, further investigation is needed for elderly subjects.

Changes in Olfactory Receptor Expression Are Correlated With Odor Exposure During Early Development in the zebrafish (Danio rerio).

We have previously shown that exposure to phenyl ethyl alcohol (PEA) causes an increase in the expression of the transcription factor otx2 in the olfactory epithelium (OE) of juvenile zebrafish, and this change is correlated with the formation of an odor memory of PEA. Here, we show that the changes in otx2 expression are specific to ßPEA: exposure to αPEA did not affect otx2 expression. We identified 34 olfactory receptors (ORs) representing 16 families on 4 different chromosomes as candidates for direct regulation of OR expression via Otx2. Subsequent in silico analysis uncovered Hnf3b binding sites closely associated with Otx2 binding sites in the regions flanking the ORs. Analysis by quantitative polymerase chain reaction and RNA-seq of OR expression in developing zebrafish exposed to different isoforms of PEA showed that a subset of ORs containing both Otx2/Hnf3b binding sites were downregulated only in ßPEA-exposed juveniles and this change persisted through adult life. Localization of OR expression by in situ hybridization indicates the downregulation occurs at the level of RNA and not the number of cells expressing a given receptor. Finally, analysis of immediate early gene expression in the OE did not reveal changes in c-fos expression in response to either αPEA or ßPEA.

Medical treatment of traumatic anosmia.

OBJECTIVES: To study the effects of zinc and steroid in the treatment of traumatic anosmia. STUDY DESIGN: A prospective, randomized study. SETTING: Academic medical center. SUBJECTS AND METHODS: Patients with a clear history of loss of smell after head injury and whose thresholds were -1 measured by the phenyl ethyl alcohol threshold test were included in this study from January 2010 to May 2013. They were randomly divided into 4 groups. Patients in group 1 were treated with zinc gluconate for a month and high-dose prednisolone with tapering for 2 weeks. Those in group 2 took only zinc gluconate, and those in group 3 took only prednisolone. Patients in group 4 did not take any medicine. All patients were followed up by phenyl ethyl alcohol threshold testing, and magnetic resonance imaging was performed to measure the volume of olfactory bulbs. RESULTS: Thirty-nine patients in group 1, 35 in group 2, 34 in group 3, and 37 in group 4 completed the study. The recovery of olfactory function was observed in 11 patients (28.2%) in group 1, in 9 (25.7%) in group 2, in 4 (11.8%) in group 3, and in 1 (2.7%) in group 4. The recovery rates of olfactory function of groups 1 and 2 were significantly higher than the recovery rate of group 4. The volume of olfactory bulbs was not significantly different between those with and without improved olfactory function. CONCLUSION: Our results show that zinc gluconate has a promising effect in treating traumatic anosmia.