Isolierung, Strukturaufklärung, Quantifizierung und Formulierung der Saponine und Flavonoide der Samen von Glinus Lotoides

Suitable extraction methods for the seeds of Glinus lotoides have been developed and the major saponins and flavonoids have been isolated. In this, four new hopane-type saponins, glinusides F, G, H and K, and the known succulentoside B as well as the two known flavones, 5,7,4'f-trihydroxyflavone-6,8-di-C-glucoside (vicenin-2) and 5,7,4'f-trihydroxy-flavone-8-C-sophoroside (vitexin-2''-O-glucoside), have been isolated from the seeds of the plant. Based on the spectral analyses including 2D NMR and HRESI mass spectroscopy, the new structures have been characterized as 3alpha-O-alpha-Dxylopyranosyl-6beta-O-alpha-D-xylopyranosyl-16alpha-O-alpha-D-xylopyranosyl-22-hydroxy-hopane (glinuside F), 3alpha-O-beta-L-rhamnopyranosyl-(1->2)-alpha-D-xylopyranosyl-6beta,16alpha-dihydroxy-22-O-beta-L-rhamnopyranosyl-hopane (glinuside G), 3alpha-O-beta-l(TM)hamnopyranosyl-( 1->2)-alpha-D-xylopyranosyl-6beta-O-alpha-D-xylopyranosyl-16alpha-hydroxy-22-O-beta-L-rhamnopyranosyl-hopane (glinuside H) and 3alpha-O-beta-L-rhamnopyranosyl-(1->2)-alpha-D-xylopyranosyl-6beta-O-alpha-D-xylopyranosyl-16alpha-O-alpha-D-xylopyranosyl-22-hopane (glinuside I). The total flavonoids and saponins of the seeds of G. lotoides in the crude extracts and tablet formulations thereof have been quantified by reversed-phase high performance liquid chromatographic (RP-HPLC) methods with UV detection. The saponins were analyzed after acid hydrolysis in 3M HCl at 100 °C for 1 hour. Vicenin-2 and mollugogenol B have been isolated and used as reference substances for the quantification of total flavonoids and saponins, respectively. The identities of vicenin-2 and mollugogenol B have been confirmed using UV, MS and NMR spectral analyses and comparison with respective published data. The purity of the isolated mollugogenol B has been determined by TLC, HPLC and UV spectrophotometry as 98%, calculated from HPLC peak area. The molar absorptivity of the methanol solution of mollugogenol B at a concentration of 0.948 mg/100ml was found to be 3.98, 4.16 and 4.12 at ƒÉmax of 260sh, 251 and 243 nm, respectively, which is in good agreement with literature values, indicating the purity of mollugogenol B. Similarly, the purity of vicenin-2 has been determined as 97% using HPLC, which is confirmed by TLC and UV methods. Satisfactory separation of the components of the flavonoids and saponins compounds has been obtained in less than 30 and 25 min, for the flavonoids and saponins, respectively. The HPLC methods were validated for linearity, repeatability, limits of detection (LOD) and quantification (LOQ). Repeatability (inter- and intra-day, n = 6 and 9, respectively) showed less than 2% relative standard deviation (RSD). The LOD and LOQ were found to be 0.075 and 0.225 mg/mL, respectively, for vicenin-2 and 0.027 and 0.082 mg/100mL, respectively, for mollugogenol B. The total amounts of saponins of the crude extract and tablet formulation have been determined as glinuside G equivalent (mol. Wt. 900) which approximates the average molecular weight of the saponins of G. lotoides. The content of flavonoids and saponins of six single tablets has been found to be between 95 and 103% for flavonoids and 94 to 98% for saponins. The validated HPLC methods could be employed to standardize a laboratory produced purified extract, which could be used as a secondary standard for the routine quality control. Accordingly, the secondary standard (extract F) has been found to contain 10% vicenin-2 and 21.3% total flavonoids calculated as vicenin-2 equivalent. The amount of saponins in the secondary standard was 6.3% mollugogenol B and 12.4% total aglycons calculated as mollugogenol B, 14.2% glinuside G and 25.4% total saponins calculated as glinuside G equivalent. Admixing of the crude liquid extract of the seeds of G. lotoides with Aeroperl(TM) 300 Pharma, as an inert carrier material, and subsequent drying the mixture has provided a non-adherent and free-flowing powder. The required amount of Aeroperl(TM) 300 Pharma has been optimized as 30%, based on the physicochemical properties such as particle size distribution, surface morphology, moisture content and sorption kinetics of the dry extract preparations containing 10, 20, 30, 40, 50 and 60% w/w Aeroperl(TM) 300 Pharma. A tablet formulation containing granules of the dry extract preparation (947 mg), Avicel(TM) PH101 (363 mg) and Ac-Di-sol (90 mg) provided the best tablet properties such as disintegration time of 2.4 min at a tablet hardness of 73 N. Granulation by roller compaction of the dry extract preparation has improved the disintegration time of the tablets. Oblong tablets which could administer the traditional dose as a single tablet have been developed and enteric coated. Enteric coated tablets of G. lotoides with pharmacopoeial requirements could be prepared using methacrylic acid/ ethylacrylate co-polymer (Eudragit(TM) L 100/55 or Kollicoat(TM) MAE 100P). Propylene glycol and acetyl tributyl citrate could be used as plasticizers at concentrations of about 11% in the dry film.Glinus lotoides wird in Athiopien traditionell als Anthelminthikum eingesetzt. Das Ziel der vorliegenden Arbeit ist es, Auszuge von G. lotoides herzustellen, diese zu charakterisieren und zu standardisieren. In einem weiteren Abschnitt sollen aus den Auszügen Arzneiformulierungen hergestellt werden. Um die Ziele der Arbeit zu erreichen, muss ein standardisiertes Extraktionsverfahren entwickelt werden. Des Weiteren ist ein umfangreiches Screening der Inhaltsstoffe durchzuführen, was die Isolierung und Identifizierung der pharmakologisch aktiven Substanzen einschliesst. Die Isolierung und die Strukturaufklarung der Saponine und der Flavonoide als wirksame Inhaltsstoffgruppen von G. lotoides liefern vier neue Hopan-Saponine, die Glinuside F, G, H und K, das bekannte Succulentosid B sowie zwei bekannte Flavonoidglykoside; 5,7,4'-trihydroxyflavon-6,8-di-C-glucosid (Vicenin-2) und 5,7,4'-trihydroxy-flavon-8-C-sophorosid (Vitexin-2"-O-glucosid). Die Strukturen der Saponine werden mittels NMR, HRESI Massenspektroskopie und Zucker-Analytik als 3beta-O-beta-D-xylopyranosyl-6alpha-O-beta-D-xylopyranosyl-16beta-O-beta-D-xylopyranosyl-22-hydroxy-hopan (Glinusid F), 3beta-O-alpha-L-rhamnopyranosyl-(1->2)-beta-D-xylopyranosyl-6alpha,16beta-dihydroxy-22-O-alpha-L-rhamnopyranosyl-hopan (Glinusid G), 3beta-O-alpha-l(TM)hamnopyranosyl-( 1->2)-beta-D-xylopyranosyl-6alpha-O-beta-D-xylopyranosyl-16beta-hydroxy-22-O-alpha-L-rhamnopyranosyl-hopan (Glinusid H) and 3beta-O-alpha-L-rhamnopyranosyl-(1->2)-beta-D-xylopyranosyl-6alpha-O-beta-D-xylopyranosyl-16beta-O-beta-D-xylopyranosyl-22-hopan (Glinusid I) aufgeklart. Die Gesamtsaponin- und Favonidgehalte der Samen von G. lotoides werden mittels RPHPLC in den Extrakten und in den Tabletten bestimmt. Die quantitative Bestimmung der Saponine wird nach saurer Hydrolyse (3M HCl, 100 °C fur 1 Stunde) durchgeführt, wobei Mollugogenol B entstehen. Für die quantitative Bestimmung der Flavonoide wird Vicenin-2 als externer Standard eingesetzt. Mittels DC und HPLC unter verschiedenen Bedingungen wird eine mindestens 97 %ige Reinheit des isolierten Vicenin-2 und Mollugogenol B gefunden. Die Berechnung der Konzentrationen erfolgt über die Peakflachen von Vicenin-2 und von Mollugogenol B nach der Methode des externen Standards. Der Gesamtflavonoidgehalt und der Gesamtsaponingehalt von G. lotoides werden über die Gesamtpeakfläche des jeweiligen Chromatogramms ermittelt. Im Rahmen der Validierung der Gehaltsbestimmungsmethode erfolgt die Bestimmung von Selektivität, Wiederfindung, System- und Methodepräzision, Bestimmungsgrenze und Nachweisgrenze. Als Sekundärstandard wird der aufgereinigte Extrakt F charakterisiert. Der Flavonoid- und Saponingehalt der Tabletten beträgt zwischen 95 und 103 % für die Flavonoide und zwischen 94 und 98 % für die Saponine. Eine zur Formulierung von Tabletten geeignete Extraktzubereitung wird durch Mischung des Extraktes mit Aeroperl(TM) 300 Pharma als Träger-Material hergestellt. Die benötigte Menge von Aeroperl(TM) 300 Pharma wird durch Untersuchungen zur Wasseraufnahme und REM-Untersuchungen optimiert. Hierbei ergibt eine Extraktzubereitung mit 30% (m/m) Aeroperl(TM) 300 Pharma die besten Eigenschaften im Hinblick auf die weitere Verarbeitung. Zusätzlich, durch Walzen-Kompaktierung der Extrakt-Aeroperl-Mischung wird die Fliessfähigkeit der Tablettiermischung und der Zerfall der Tabletten verbessert. Eine geeignete Tablettenformulierung, die 947 mg Extraktzubereitung, 363 mg Avicel(TM) PH101, und 90 mg Ac-Di-sol enthält, wird durch mehrere Optimierungsschritte entwickelt. Diese Tabletten zeigen einen Zerfall der Tabletten innerhalb von 2,4 Minuten bei einer Harte von 73 N. Zusätzlich erhalten die Oblong-Tabletten einen Magensaftresistenten Überzug, der den Arzneibuchanforderungen entspricht. Dieser Überzug wird mittels der hier für geeigneten Filmbildner Eudragit(TM) L 100/55 oder Kollicoat(TM) MAE 100P hergestellt. Der Filmüberzug enthält Propylenglykol und Acetyltributylcitrat als Weichmacher. Durch die Prüfung der Zerfallzeit wird sichergestellt, dass die überzogenen Tabletten in künstlichen Magensaft Ph. Eur. (0,1 M Salzsäure, pH 1,0) innerhalb 2 Stunden nicht zufallen jedoch anschliessend in künstlichen Darmsaft (Phosphat-Pufferlosung, pH 6,8) innerhalb von 15 Minuten zufallen. Zusammenfassend kann festgestellt werden, dass es gelungen ist, einen Auszug von G. lotoides herzustellen, dieser zu einer zur Formulierung von Tabletten geeigneten Extraktzubereitung zu verarbeiten, eine Tablettenformulierung zu entwickeln und die Tabletten mit einem magensaftresistenten Überzug zu versehen

Проблемы и перспективы вступления Эфиопии в ВТО

The article is devoted to the analysis of problems and prospective of Ethiopia's joining WTO. The advantages and disadvantages of the process, its costs and benefits are considered. The author takes into account that the country is in the stage of infant industrialization of its economic development. One of the paper's conclusions is that the industrialization of their economy is of vital necessity for least developed countries, which should be taken into account in their aspiration to become WTO members. The general conclusion is that WTO membership of least developed countries including Ethiopia may have a negative consequence in the short and medium term and positive consequence in the long-term development.Статья посвящена анализу проблем и перспектив присоединения Эфиопии к ВТО. Рассматриваются преимущества и трудности этого процесса, затраты и выгоды. Автор учитывает то обстоятельство, что страна находится на начальном этапе своего индустриального развития. Наименее развитые страны должны делать упор на индустриализацию экономики. Общий вывод состоит в том, что для наименее развитых стран, включая Эфиопию, членство в ВТО может иметь отрицательные последствия для их экономического развития в краткосрочной и среднесрочной перспективе и положительные в долгосрочной перспективе

Brain magnetic resonance imaging findings in patients with developmental delay in Addis Ababa, Ethiopia

BACKGROUND: Developmental delay is a major health problem throughout the world causing significant individual disability. Even though physical examination and patient history are the most important and basic evaluations of patients with developmental delay, additional investigations are usually required in supporting or reaching a diagnosis among which is neuroimaging. This study aims to assess brain Magnetic resonance imaging (MRI) patterns in patients presented with developmental delay.METHOD: A retrospective analysis of 164 patients who had undergone brain Magnetic Resonance Imaging (MRI) evaluation for the developmental delay was done. The study was conducted between March to November 2021 G.C at Tikur Anbessa Specialized Hospital (TASH). The patients’ clinical history and magnetic resonance imaging reports were reviewed from their medical records. All patients with developmental delay who had brain MRI evaluation at TASH and at one private diagnostic center in Addis Ababa were included in the study.RESULTS: A total of 164 patients were included in this study of which 95(57.9%) were male and 69(42.1%) female patients were seen. A total of 120 patients (73.2%) showed abnormal brain MRI studies. Previous neurovascular insults were the most common abnormalities seen in 75(45.7%) patients followed by imaging findings of congenital and developmental abnormalities seen in 20(12.2%) patients.CONCLUSION: Brain MRI is an important input in the evaluation of patients with developmental delay. It can give evidence for the cause of developmental delay, especially in the diagnosis of perinatal/hypoxic-ischemic insults, and congenital and developmental malformations

Breast cancer in Ethiopia: evidence for geographic difference in the distribution of molecular subtypes in Africa

Abstract Background Breast cancer is a heterogeneous disease with several morphological and molecular subtypes. Widely accepted molecular classification system uses assessment of estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 (HER2) and proliferation marker Ki67. Few studies have been conducted on the incidence and molecular types of breast cancer in Sub-Saharan Africa. Previous studies mainly from Western and Central Africa, showed breast cancer to occur at younger ages and to present with aggressive features, such as high-grade, advanced stage and triple-negative phenotype (negative for ER, PR and HER2). Limited data from East Africa including Ethiopia however shows hormone receptor negative tumors to account for a lower proportion of all breast cancers than has been reported from elsewhere in Africa. Methods In this study from Tikur Anbessa Specialized Hospital, 114 breast cancer patients diagnosed between 2012 and 2015 were enrolled. ER, PR, Ki67 and HER2 receptor status were assessed using immunohistochemistry from tissue microarrays. FISH was used for assessment of gene amplification in all equivocal tumor samples and for confirmation in HER2-enriched cases. Results The distribution of molecular subtypes was: Luminal A: 40%; Luminal B: 26%; HER2-enriched: 10%; TNBC: 23%. ER were positive in 65% of all tumors and 43% the cases were positive for PR. There was statistically significant difference in median age at diagnosis between the molecular subtypes (P < 0.05). There was a bimodal distribution of molecular subtypes in different age ranges with Luminal B subtype being more common at younger ages (median = 36) and Luminal A subtype more prevalent at older ages (median = 42). There were no statistically significant differences in tumor grade, histology, and stage between the molecular subtypes of breast cancer. Conclusion The present study detected Luminal A breast cancer to be the most common subtype and reveals a relatively low rate of hormone receptor negative and TNBC. Our findings and results from other East African studies suggest geographic variability in the distribution of the molecular subtypes of breast cancer in Africa and hence have important clinical and policy implications for breast cancer control and treatment in Ethiopia.https://deepblue.lib.umich.edu/bitstream/2027.42/142374/1/12905_2018_Article_531.pd

The Effects of Prednisolone Treatment on Cytokine Expression in Patients with Erythema Nodosum Leprosum Reactions.

Erythema nodosum leprosum (ENL) is a systemic inflammatory complication occurring mainly in patients with lepromatous leprosy (LL) and borderline lepromatous leprosy. Prednisolone is widely used for treatment of ENL reactions but clinical improvement varies. However, there is little good in vivo data as to the effect of prednisolone treatment on the pro-inflammatory cytokines in patients with ENL reactions. As a result, treatment and management of reactional and post-reactional episodes of ENL often pose a therapeutic challenge. We investigated the effect of prednisolone treatment on the inflammatory cytokines TNF, IFN-γ, IL-1β, IL-6, and IL-17 and the regulatory cytokines IL-10 and TGF-β in the skin lesion and blood of patients with ENL and compared with non-reactional LL patient controls. A case-control study was employed to recruit 30 patients with ENL and 30 non-reactional LL patient controls at ALERT Hospital, Ethiopia. Blood and skin biopsy samples were obtained from each patient before and after prednisolone treatment. Peripheral blood mononuclear cells from patients with ENL cases and LL controls were cultured with M. leprae whole-cell sonicates (MLWCS), phytohemagglutinin or no stimulation for 6 days. The supernatants were assessed with the enzyme-linked immunosorbent assay for inflammatory and regulatory cytokines. For cytokine gene expression, mRNA was isolated from whole blood and skin lesions and then reverse transcribed into cDNA. The mRNA gene expression was quantified on a Light Cycler using real-time PCR assays specific to TNF, IFN-γ, IL-β, TGF-β, IL-17A, IL-6, IL-8, and IL-10. The ex vivo production of the cytokines: TNF, IFN-γ, IL-1β, and IL-17A was significantly increased in untreated patients with ENL. However, IL-10 production was significantly lower in untreated patients with ENL and significantly increased after treatment. The ex vivo production of IL-6 and IL-8 in patients with ENL did not show statistically significant differences before and after prednisolone treatment. The mRNA expression in blood and skin lesion for TNF, IFN-γ, IL-1β, IL-6, and IL-17A significantly reduced in patients with ENL after treatment, while mRNA expression for IL-10 and TGF-β was significantly increased both in blood and skin lesion after treatment. This is the first study examining the effect of prednisolone on the kinetics of inflammatory and regulatory cytokines in patients with ENL reactions before and after prednisolone treatment. Our findings suggest that prednisolone modulates the pro-inflammatory cytokines studied here either directly or through suppression of the immune cells producing these inflammatory cytokines

Global, regional, and national burden of disorders affecting the nervous system, 1990–2021: a systematic analysis for the Global Burden of Disease Study 2021

BackgroundDisorders affecting the nervous system are diverse and include neurodevelopmental disorders, late-life neurodegeneration, and newly emergent conditions, such as cognitive impairment following COVID-19. Previous publications from the Global Burden of Disease, Injuries, and Risk Factor Study estimated the burden of 15 neurological conditions in 2015 and 2016, but these analyses did not include neurodevelopmental disorders, as defined by the International Classification of Diseases (ICD)-11, or a subset of cases of congenital, neonatal, and infectious conditions that cause neurological damage. Here, we estimate nervous system health loss caused by 37 unique conditions and their associated risk factors globally, regionally, and nationally from 1990 to 2021.MethodsWe estimated mortality, prevalence, years lived with disability (YLDs), years of life lost (YLLs), and disability-adjusted life-years (DALYs), with corresponding 95% uncertainty intervals (UIs), by age and sex in 204 countries and territories, from 1990 to 2021. We included morbidity and deaths due to neurological conditions, for which health loss is directly due to damage to the CNS or peripheral nervous system. We also isolated neurological health loss from conditions for which nervous system morbidity is a consequence, but not the primary feature, including a subset of congenital conditions (ie, chromosomal anomalies and congenital birth defects), neonatal conditions (ie, jaundice, preterm birth, and sepsis), infectious diseases (ie, COVID-19, cystic echinococcosis, malaria, syphilis, and Zika virus disease), and diabetic neuropathy. By conducting a sequela-level analysis of the health outcomes for these conditions, only cases where nervous system damage occurred were included, and YLDs were recalculated to isolate the non-fatal burden directly attributable to nervous system health loss. A comorbidity correction was used to calculate total prevalence of all conditions that affect the nervous system combined.FindingsGlobally, the 37 conditions affecting the nervous system were collectively ranked as the leading group cause of DALYs in 2021 (443 million, 95% UI 378–521), affecting 3·40 billion (3·20–3·62) individuals (43·1%, 40·5–45·9 of the global population); global DALY counts attributed to these conditions increased by 18·2% (8·7–26·7) between 1990 and 2021. Age-standardised rates of deaths per 100 000 people attributed to these conditions decreased from 1990 to 2021 by 33·6% (27·6–38·8), and age-standardised rates of DALYs attributed to these conditions decreased by 27·0% (21·5–32·4). Age-standardised prevalence was almost stable, with a change of 1·5% (0·7–2·4). The ten conditions with the highest age-standardised DALYs in 2021 were stroke, neonatal encephalopathy, migraine, Alzheimer's disease and other dementias, diabetic neuropathy, meningitis, epilepsy, neurological complications due to preterm birth, autism spectrum disorder, and nervous system cancer.InterpretationAs the leading cause of overall disease burden in the world, with increasing global DALY counts, effective prevention, treatment, and rehabilitation strategies for disorders affecting the nervous system are needed

Problems and Prospective for Ethiopia's Joining WTO

The article is devoted to the analysis of problems and prospective of Ethiopia's joining WTO. The advantages and disadvantages of the process, its costs and benefits are considered. The author takes into account that the country is in the stage of infant industrialization of its economic development. One of the paper's conclusions is that the industrialization of their economy is of vital necessity for least developed countries, which should be taken into account in their aspiration to become WTO members. The general conclusion is that WTO membership of least developed countries including Ethiopia may have a negative consequence in the short and medium term and positive consequence in the long-term development

The place of children through the lens of generational and gender relationships in Awra Amba community, northern Ethiopia

This study explores the place of children through the lenses of generational and gender relationships in Awra Amba community, Northern Ethiopia. It discusses what children’s life from gender and generational perspective look like in Awra Amba community. The study is based on the following research questions. What are the social values and roles of children in the community? What does gender relationship among children and between children and adults look like in Awra Amba community? What is “special” about children of Awra Amba Community? To document the place of children through answering these questions, the study employs qualitative research methods such as in-depth interview, focus group discussion observation, informal dialogue and home visit. These qualitative methods were used as the issue to be explored needed a complex and detailed understanding. In other words, the research project was qualitative in its nature because the researcher needed an intricate and thorough understanding of the issue. Children whose age ranges from12-15 years and their parents (both mothers and fathers) whose age ranges from 32-40 and 48-72 respectively including the founder and the leader of the community, Zumra Nuru, were included in the study as participants (research subjects). The study is informed by social constructivism through viewing children’s place and childhood among the Awura Amba community from discursive construction, structural construction, generational construction and children as social actors perspectives. The study observes children’s place among the community from three angles: gender equality, social order and intergenerational relationships. It argues that children are active participants in activation and perpetuation of gender equity, social order and egalitarian intergenerational relationships. Such perspectives of children indicate that children are well aware of the collective values among the community and are exercising them as part of their everyday lives

Utilization of post abortal contraceptive use and associated factors among women who came for abortion service at Debre Berhan Hospital, Debre Berhan, Ethiopia March 2019: Institution based cross sectional study

Background: Post abortion family planning (PAFP) is the initiation and use of family planning methods immediately after, and within 48 hours of an abortion, before fertility returns. In most women fertility returns on average about two weeks after an abortion; however, ovulation can occur as early as 11 days post-abortion. Objective: To assess utilization of post abortal contraceptive use and associated factors among women who came for abortion service at Debre Berhan Referral Hospital, Debre Berhan, Ethiopia March 2019. Methodology: Institutional based cross sectional study design was conducted using hospital data obtained from Gynecology ward in Debre Berhan Referral Hospital, Debre Berhan, Ethiopia, from March 1 -10, 2019. A systematic random sampling technique was used from the abortion register log book. Data was cleaned manually, coded and entered into Epi-data version 3.1 then exported to and analyzed by SPSS version 21 software. Multivariate analysis with AOR, 95% CI and p-value&lt; 0.05 were used to identify variables which have significant association. Result: The finding of the current study showed that among 371 study subjects 170(45.8%) utilized post abortal family planning. There was a significant association between utility of post abortal family planning and post abortion family planning counseling [AOR: 19.245, 95% CI: (10.199, 36.313), p-value= 0.001] and women who were primiparous had 5 times more likely to utilize post abortal family planning as compared to the women who were nullyparous [AOR: 5.314, 95%CI (1.089, 24.210), p value=0.001]. Conclusion and Recommendation: From a total of 371 study subjects 45.8% have utilize contraceptive after abortion service received. This study also showed that parity and counseling&rsquo;s of family planning were statistically significant associated with utilization of post abortal family planning. We recommend Debre Berhan hospital to scale up activities on post abortal care to increase the number of clients who post abortal family planning