Bioassay guided isolation of compounds with acetylcholinesterase inhibitory activity from selected medicinal plants used in Iranian traditional medicine

Kognitive Störungen sind eine der Ursachen von mentalem Abbau, der von Symptomen wie Erinnerungsverlust und Gedächnisschwund z.b. im Rahmen der Alzheimer-Krankheit begleitet wird. Die Steigerung von cholinergen Funktionen durch Inhibierung von Acetylcholinesterase (AChE) stellt eine therapeutische Möglichkeit zur Behandlung von neurologischen Störungen wie Alzheimer-Krankheit und Demenz dar. In den letzten zwei Jahrzehnten rückte auch der Einsatz traditioneller Heilpflanzen zur Therapie von Demenz-Krankheiten in den Mittelpunkt des wissenschaftlichen Interesses. Der Iran ist eine der Kulturen mit einer langen und reichen Geschichte der traditionellen Medizin, und speziell der Pflanzenheilkunde. In dieser Studie wurden 40 Heilpflanzen bzw. Pflanzenprodukte wie Harze, die in der Iranischen Traditionellen Medizin zur Behandlung von kognitiven Störungen eingesetzt wurden, untersucht. Zuerst wurden polare methanolische und apolare Dichlormethan-Extrakteaus den selektierten Pflanzen hergestellt. Die Aktivität der Extrakte, AChE zu hemmen, wurde mittels Dünnschichtchromatographie und in einem Microplate Assay analysiert. Die stärkste Inhibierungs wurde für Extrakte aus den Samen von Peganum harmalaund aus Harzen von Semecarpus anacardium, Dorema ammoniacum und Ferula gummosa festgestellt. Diese vier Ausgangsmaterialien wurden für die weiteren Analysen verwendet. Die aktiven Inhaltsstoffe wurden mittels chromatographischer Methoden wie Vakuumflüssigchromatographie, Säulenchromatographie, Gel-Permeations-Chromatographie, Festphasenextraktion, Gegenstromverteilungschromatographie und automatisierter HPTLC isoliert. Die Charakterisierung der isolierten bioaktiven Komponten erfolgte durch ein- und zwei dimensionale 1H and 13C NMR Spektroskopie (COSY, TOCSY, HSQC, HMBC, NOESY) und Massenpektrometrie. Die Hemmwirkung der aktiven Extrakte und isolierten Verbindungen auf AChE wurde durch die Bestimmung der IC50 Werte quantifiziert. Ausserdem wurden die Konzentrationen dieser Wirkkomponenten in den einzelnen Drogen mittels validierter HPLC-Verfahren bestimmt. In Samen von Peganum harmala wurden die Alkaloide Harmine und Harmaline als die aktiven Komponente identifiziert. Die Korrelation zwischen den IC50 Werten der Extrakte und den Konzentrationen der aktiven Komponenten zeigte, dass die Hemmung der AChE durch Peganum harmala überwiegend auf diese zwei Alkaloide zurückzuführen ist. Im Harz der Früchte von Semecarpus anacardium wurden 1’,2’-dihydroxy-3’-pentadec-8-enylbenzene und 1’,2’-dihydroxy-3’-pentadeca-8,11-dienylbenzene als Komponenten mit inhibitorischer Wirkung auf AchE identifiziert. Dieser Effekt konnte zusätzlich durch ein in Silico Experiment bestätigt werden. Weitere Experimente zeigten, dass diese Komponenten nur AChE inhibieren, während sie keine Hemmung der Butyrylcholinesterase (BchE) bewirkten. Zusätzlich konnte eine Korrelation zwischen der AchE Inhibierung und der Konzentration dieser Substanzen im Harz nachgewiesen werden. Diese Resultate deuten erstmalig darauf, dass diese zwei Substanzen für die AChE Inhibierung durch das Harz der Früchte von Semecarpus anacardium hauptverantwortlich sind. Im Harz von Dorema ammoniacum wurden 2 Inhaltstoffe (2'S,5'S)-2'-ethenyl-5'-(3-hydroxy-6-methyl-4-oxohept-5-en-2-yl)-7-methoxy-2'-methyl-4H-spiro[chromene-3,1'-cyclopentane]-2,4-dione und (2'S,5'R)-2'-ethenyl-5'-[(2R,4R)-4-hydroxy-6-methyl-3-oxohept-5-en-2-yl]-7-methoxy-2'-methyl-4H-spiro[chromene-3,1'-cyclopentane]-2,4-dione mit AChE-hemmender Wirkung identifiziert. Der zweite Inhaltstoff ist ein neuer Naturstoff, der zum ersten Mal in dieser Arbeit identifiziert werden konnte. Beide Inhaltstoffe waren in relativ hoher Konzentration im Harz enthalten und die Korrelation zwischen den Konzentrationen und dem Inhibierungseffekt war wiederum ein starkes Indiz dafür, dass diese Inhaltstoffe für die AchE Inhibierung verantwortlich sind. Aus dem Harz von Ferula gummosa wurden die Komponenten 7-{[(2E)-3,7-dimethylocta-2,6-dien-1-yl]oxy}-2H-chromen-2-one und 7-(((1R,4aR,6S,8aS)-6-hydroxy-5,5,8a-trimethyl-2-methylenedecahydronaphthalen-1-yl)methoxy)-2H-chromen-2-one isoliert und charakterisiert. Die relativ hohen Konzentrationen dieser Komponenten in Korrelation mit der AChE-Inhibierung lassen die Anwendung dieses Harzes in der Iranischen traditionellen Medizin plausibel erscheinen. Diese Studie zeigte ausserdem, dass die HPTLC-MS/NMR als eine sehr effiziente Methode zur direkten und schnellen Identifizierung von Inhaltstoffen aus komplexen Pflanzenextrakten herangezogen werden kann, wobei eine optimierte Trennung in der HPTLC eine wichtige Voraussetzung ist. Im Rahmen dieser Dissertation wurde zum ersten Mal die Aktivität der Inhaltstoffe von vier Heilpflanzen bzw. Pflanzenprodukten aus der Iranischen Traditionellen Medizin auf AChE-Hemmung nachgewiesen. Die Isolierung, Charakterisierung und Bestimmung der IC50 Werte dieser Inhaltstoffe wurde erfolgreich durchgeführt. Durch diese Arbeit konnte der Einsatz dieser Heilpflanzen in der Iranischen Traditionellen Medizin zur Behandlung von kognitiven Störungen wissenschaftlich belegt werden.Cognitive deficits are one of the most common causes of mental deterioration such as learning impairment, delayed amnesia and memory problems e.g. in Alzheimer’s disease. The enhancement of cholinergic function by inhibition of acetylcholinesterase (AChE) is considered as a rational approach for the treatment of neurological disorders such as Alzheimer´s disease and senile dementia. During the last two decades the use of herbal medicinal substances in dementia therapy has been studied well. Iran is among those countries with the longest and richest history in Traditional Medicine and especially in the use of herbal medicinal preparations. In this thesis 40 herbal drugs or plant products that are reported in Iranian traditional medicine (ITM) or ethnomedicine as sources for the treatment of cognitive disorders were examined. In the first step, the bioactivity of polar methanolic and non-polar dichloromethane extracts of all selected herbal samples from ITM was studied on AChE inhibition by TLC bioautography and in a microplate assay. The most active herbal drugs were the seeds from Peganum harmala, the fruit resin from Semecarpus anacardium, the gum-resin from Dorema ammoniacum and the oleo gum-resin from Ferula gummosa. Those were selected for further study. In detailed investigations, the active compounds were identified and isolated using several chromatographic techniques such as thin layer chromatography, vacuum liquid chromatography, column chromatography, size exclusion chromatography, solid phase extraction, high performance liquid chromatography, counter current chromatography and automated high performance thin layer chromatography. The structures of the active components were characterized by one and two-dimensional 1H and 13C NMR spectroscopy (COSY, TOCSY, HSQC, HMBC, NOESY) and mass spectrometry. The IC50 values for active compounds were determined by a quantitative colorimetric assay. Additionally, the concentrations of active components in the four different sources were determined by HPLC analysis. The active compounds in the seeds of Peganum harmala were identified as harmine and harmaline. From the correlation of IC50 values of the extracts in AChE inhibition with the concentrations of harmaline and harmine in the respective extracts can be concluded that these two alkaloids are the major AChE inhibitory compounds in Peganum harmala. The characterization of the active compounds in the fruit resin of Semecarpus anacardium resulted in 1’,2’-dihydroxy-3’-pentadec-8-enylbenzene and 1’,2’-dihydroxy-3’-pentadeca-8,11-dienylbenzene. In addition, an in silico study confirmed their AChE inhibitory effect. Further experiments showed their selective inhibitory activity for AChE versus BChE. The correlation of AChE inhibition with the percentage of the active compounds in the fruit resin, can explain their responsibility for the AChE inhibition of this resin. In the gum-resin of Dorema ammoniacum the active components were identified as (2'S,5'S)-2'-ethenyl-5'-(3-hydroxy-6-methyl-4-oxohept-5-en-2-yl)-7-methoxy-2'-methyl-4H-spiro[chromene-3,1'-cyclopentane]-2,4-dione and (2'S,5'R)-2'-ethenyl-5'-[(2R,4R)-4-hydroxy-6-methyl-3-oxohept-5-en-2-yl]-7-methoxy-2'-methyl-4H spiro[chromene-3,1'-cyclopentane]-2,4-dione. The second compound is a new natural substance. From the high concentration of these two components in this gum-resin can be deduced they are of among the major compounds responsible for the AChE inhibitory activity of this drug. The structure elucidation of the active substances in the oleo gum-resin of Ferula gummosa gave 7-{[(2E)-3,7-dimethylocta-2,6-dien-1-yl]oxy}-2H-chromen-2-one and 7-(((1R,4aR,6S,8aS)-6-hydroxy-5,5,8a-trimethyl-2-methylenedecahydronaphthalen-1-yl)methoxy)-2H-chromen-2-one. This is the first report of these two compounds in F. gummosa and from the result can be concluded that they are of importance for the activity. The study showed that HPTLC-MS/NMR can be considered as a fast method for dereplication of natural compounds, when an optimum separation on HPTLC plate is provided. The AChE inhibition of the isolated compounds from the investigated herbal drugs and their IC50 values were reported for the first time in this thesis. The achieved results confirmed that the compounds considerably contribute to the effects of these drugs and underline the plausibility of their use in the treatment of cognitive deficits in ITM

Evaluation of Patient Satisfaction Referred to Health Center Traditional Medicine in Shahid Beheshti University of Medical Sciences in 2016

زمینه و هدف: رضایتمندی از خدمات سلامت یک معیار مهم برای سنجش کیفیت مراقبت‌های بهداشتی شناخته شده است و تاکنون بررسی از نظر میزان رضایتمندی بیماران مراجعه‌کننده به سلامتکده‌های طب سنتی انجام نشده است، لذا در این مطالعه برای نخستین‌بار به سنجش کیفیت خدمات در سلامتکده طب سنتی دانشگاه شهید بهشتی پرداخته شد. مواد و روش‌ها: مطالعه حاضر یک مطالعه توصیفی ـ مقطعی (Cross–Sectional) است که جهت سنجش رضایت بیماران، از 150 بیمار مراجعه‌کننده به سلامتکده طب سنتی دانشگاه شهید بهشتی در سال 1395 انجام شد. نمونه‌ها از طریق نمونه‌گیری در دسترس، پس از اخذ رضایت‌نامه، با پرسشنامه رضایتمندی محقق‌ساخته بررسی شدند، سپس اطلاعات جمع‌آوری‌شده توسط نرم‌افزار SPSS 20 و شاخص‌های آماری توصیفی و استنباطی مورد تجزیه و تحلیل قرار گرفت. ملاحظات اخلاقی: این پژوهش حاصل طرح تحقیقاتی مصوب معاونت پژوهش دانشگاه علوم پزشکی شهید بهشتی با کد IR.SBMU.RETECH.REC.1395.487 می‌باشد و کلیه اطلاعات بیماران به صورت محرمانه حفظ شد. یافته‫ها: میزان رضایتمندی بیماران نسبت به عملکرد کلی سلامتکده خوب (میانگین 4/54) گزارش شد. میانگین امتیاز رضایتمندی در ابعاد مختلف، شامل وضعیت برخورد پزشکان با بیماران 4/73، وضعیت برخورد دستیاران با بیماران 4/69، وضعیت فضا و امکانات بهداشتی 4/66، وضعیت خدمات تشخیصی 4/74، وضعیت خدمات درمانی 4/50، نحوه وقت‌دهی و برخورد پرسنل 4/59، رضایت از داروخانه 4/62 که نشان‌دهنده طیف نسبتاً راضی به سمت کاملاً راضی بوده است. میانگین بیشترین میزان رضایت در بین بخش‌های مختلف تخصصی، از نظر وضعیت برخورد پزشکان با بیماران مربوط به بخش گوارش 4/82 می‌باشد. بیشترین میزان رضایت مراجعه‌کنندگان در بین بخش‌های مختلف تخصصی از نظر وضعیت خدمات تشخیصی (بخش مفاصل، زنان، گوارش و اعصاب و روان) مربوط به بخش اعصاب و روان بود (P=0/039). نتیجه‌گیری: رضایت بیماران از ارائه خدمات سلامتکده مورد مطالعه در سطح مطلوبی قرار داشت، اما در جهت ارائه خدمات با کیفیت بالاتر به بیماران انجام برنامه‌ریزی دقیق‌تر ضروری به نظر می‌رسد.Background and Aim: Satisfaction from health services is an important criterion for measuring the quality of health care and so far, there has been no study on the satisfaction of patients referring to traditional medicine health centers. Therefore, in this study, for the first time, the quality of services in the traditional medicine health center of Shahid Beheshti University was assessed. Materials and Methods: The present study is a cross-sectional study, in order to measure patients satisfaction, out of 150 patients referred to Shahid Beheshti University Traditional Medicine Health Center, it was performed in 2016. Samples were examined through available sampling, after obtaining consent, with the researcher-made satisfaction questionnaire. Data were analyzed by SPSS 20 software and descriptive and inferential statistical indices. Ethical Considerations: This research is the result of a research project approved by the Research Vice Chancellor of Shahid Beheshti University of Medical Sciences with the code IR.SBMU.RETECH.REC.1395.487 and all patient information was kept confidential. Findings: Patients' satisfaction with good general health center performance was reported (Mean 4/54). Mean of satisfaction in different dimensions, including The situation of doctors with patients was 4/73, the situation of patients with patients 4/69, the condition of space and health facilities 4/66, the status of diagnostic services, 4/74, the state of health care 4/40, the manner of staffing and staffing 4/59, satisfaction from the pharmacy 4/62 that this range represents a fairly pleasingly satisfactory range. The average of the highest satisfaction among different specialized departments, in terms of the situation of doctors, with patient’s gastrointestinal tract are 4/82. The highest satisfaction rate was observed in the diagnostic services status in the various specialized departments, which is related to the neurology and psychiatry is significant (P=0/039). Conclusion: Patients' satisfaction from the health services provided by the study center was at a desirable level .In order to provide higher quality services, patients need to be more accurately planned.   Please cite this article as: Tansaz M, Adhami S, Choopani R, Mokaberinejad R, Movahhed M, Ghasemi E, Ilkhani R. Evaluation of Patient Satisfaction Referred to Health Center Traditional Medicine in Shahid Beheshti University of Medical Sciences in 2016. Med Hist J 2020; 12(43): 61-72

Classifying Mammographic Mass Shapes Using the Wavelet Transform Modulus-maxima Method

In this article, multiresolution analysis, specifically the discrete wavelet transform modulus-maxima (mod-max) method, is utilized for the extraction of mammographic mass shape features. These shape features are used in a classification system to classify masses as round, nodular, or stellate. The multiresolution shape features are compared with traditional uniresolution shape features for their class discriminating abilities. The study involved 60 digitized mammographic images. The masses were segmented manually by radiologists, prior to introduction to the classification system. The uniresolution and multiresolution shape features were calculated using the radial distance measure of the mass boundaries. The discriminating power of the shape features were analyzed via linear discriminant analysis (LDA). The classification system utilized a simple Euclidean metric to determine class membership. The system was tested using the apparent and leave-one-out test methods. The classification system when using the multiresolution and uniresolution shape features resulted in classification rates of 83% and 80% for the apparent and leave one-out test methods, respectively. In comparison, when only the uniresolution shape features were used, the classification rates were 72 and 68% for the apparent and leave-one-out test methods, respectively

Feature Point Extraction from the Local Frequency Map of an Image

We propose a novel technique for detecting rotation- and scale-invariant interest points from the local frequency representation of an image. Local or instantaneous frequency is the spatial derivative of the local phase, where the local phase of any signal can be found from its Hilbert transform. Local frequency estimation can detect edge, ridge, corner, and texture information at the same time, and it shows high values at those dominant features of an image. For each pixel, we select an appropriate width of the window for computing the derivative of the phase. In order to select the width of the window for any given pixel, we make use of the measure of the extent to which the phases, in the neighborhood of that pixel, are in the same direction. The local frequency map, thus obtained, is then thresholded by employing a global thresholding approach to detect the interest or feature points. Repeatability rate, a performance evaluation criterion for an interest point detector, is used to check the geometric stability of the proposed method under different transformations. We present simulation results of the detection of feature points from image utilizing the suggested technique and compare the proposed method with five existing approaches that yield good results. The results prove the efficacy of the proposed feature point detection algorithm. Moreover, in terms of repeatability rate; the results show that the performance of the proposed method with respect to different aspect is compatible with the existing methods