Mobility-shift-Affinitätskapillarelektrophorese zur Untersuchung von Protein-Metallionen-Wechselwirkungen: Aspekte der Methodenentwicklung, -validierung und des High-Throughput-Screenings

Approximately one third of all proteins are metalloproteins, which means the metal ions are important for their biological functions. Therefore, investigation and characterization of interactions between proteins and metal ions are interesting. Several techniques can be used for these purposes. Mobility shift-affinity capillary electrophoresis has been selected over the other techniques due to its rapid analysis, small injection volume (nano-level), high separation efficiency, direct injection of impure samples and the ability to perform under physiological-like conditions. In order to facilitate the implementation of a mobility shift-affinity capillary electrophoresis method into routine binding studies, method acceleration, transfer and precision improvement were investigated. The mobility shift-affinity capillary electrophoresis method was successfully accelerated by using shorter capillaries, lower sample concentrations and smaller injection volumes. Fast run time of 4 min has been achieved. The precision of results was enhanced by slightly pushing the sample into the capillary, refreshing the buffer vials after each 30 runs and employing extra flushing after each 60 subsequent runs and using 0.1 M EDTA within the rinsing protocols. The accelerated mobility shift-affinity capillary electrophoresis method has been successfully applied for the interaction study of Li+, Na+, Mg2+, Ca2+, Ba2+, Al3+, Ga3+, La3+, Pd2+, Ir3+, Ru3+, Rh3+, Pt2+, Pt4+, Os3+, Au3+, Au+, Ag+, Cu+, Cu2+, Fe2+, Fe3+, Co2+, Ni2+, Cr3+, V3+, MoO42- and SeO32- with bovine serum albumin, β-lactoglobulin, human serum albumin, myoglobin and ovalbumin. Excellent precision for mobility ratios was achieved for all protein-metal ion interactions (RSD% of 0.05-1.0%, n > 324). The calculated normalized difference of mobility ratio and its sign (positive and negative) for each protein-metal ion interaction have been successfully used to detect the interaction and estimate further coordination of the bound metal ion, respectively. Furthermore, the results were successfully discussed in view of the properties of the metal ions and proteins, and the HSAB concept as well. The comprehensive platform summarizes all the obtained interaction results, and is valuable for any future protein-metal ion investigation.Schätzungsweise ein Drittel aller Proteine sind Metalloproteine, dies bedeutet, dass Metallionen für ihre biologische Funktion von Bedeutung sind. Somit ist die Untersuchung und Charakterisierung der Wechselwirkungen zwischen Proteinen und Metallionen von Interesse. Einige Techniken können für diesen Zweck verwendet werden. Der Mobility-shift-Affinitätskapillarelektrophorese wurde gegenüber anderen Techniken der Vorzug gegeben, da sie kurze Analysenzeiten, Injektionen von geringen Probenvolumina (im Nanoliterbereich), hohe Trennungseffizienz, direkte Injektion von nicht aufgearbeitet Proben und die Möglichkeit unter physiologischen Bedingungen zu arbeiten vereint. Um das Einbinden einer Mobility-shift-Affinitätskapillarelektrophoresemethode in Routinebindungsstudien zu erleichtern, wurden eine Verkürzung der Analysendauer, der Methodentransfer und die Verbesserung der Präzision untersucht. Die Mobility-shift-Affinitätskapillarelektrophoresemethode wurde erfolgreich beschleunigt durch Verkürzen der Kapillare, Verwenden geringerer Probenkonzentrationen und geringerer Probenvolumina. Hierbei wurden kurze Analysezeiten von 4 Minuten erreicht. Die Präzision der Ergebnisse wurden durch ein leichtes Drücken der Probe in die Kapillare nach der Injektion, das Tauschen der Puffergefäße nach 30 Läufen, Einführen eines weiteren Spülschrittes nach 60 aufeinanderfolgenden Läufen und Verwendung von 0.1 M EDTA-Lösungen im Spülprotokoll verbessert. Diese verbesserte Mobility-shift-Affinitätskapillarelektrophoresemethode wurde erfolgreich verwendet um Interaktionsstudien zwischen Li+, Na+, Mg2+, Ca2+, Ba2+, Al3+, Ga3+, La3+, Pd2+, Ir3+, Ru3+, Rh3+, Pt2+, Pt4+, Os3+, Au3+, Au+, Ag+, Cu+, Cu2+, Fe2+, Fe3+, Co2+, Ni2+, Cr3+, V3+, MoO42- und SeO32- mit bovinem Serumalbumin, β-Lactoglobulin, humanem Serumalbumin, Myoglobin und Ovalbumin. Ausgezeichnete Präzision wurde für den Mobilitätsquotienten jeder Protein-Metallionen-Interaktion (RSD% von 0,05-1,0%, n > 324). Die berechnete, normalisierte Differenz der Mobilitätsquotienten und deren Vorzeichen (positiv und negativ) jeder Protein-Metalionen-Interaktion konnten erfolgreich verwendet werden um Interaktionen zu erkennen und die weitere Koordination der Metallionen abzuschätzen. Des Weiteren konnten die Ergebnisse in Hinblick auf die Eigenschaften der Metallionen und Proteine sowie des HSAB-Konzepts erfolgreich diskutiert werden. Die Gesamtübersicht fasst alle erhaltenen Interaktionsergebnisse zusammen und ist wertvoll für zukünftige Protein-Metallionenuntersuchungen

Synthesis of some novel pyrimidine, thiophene, coumarin, pyridine and pyrrole derivatives and their biological evaluation as analgesic, antipyretic and anti-inflammatory agents

Pyrimidine derivative 3 was afforded through the reaction of compound (1) with 5-ureidohydantion (2). Product 3 underwent a cyclization to produce fused pyrimidine derivative 7, although the latter product 7 was synthesized through one step via the reaction of compound (1) with 5-ureidohydantion (2) using another catalyst. Compound 3 was oriented to react with cyclic ketones 8a,b in the presence of elemental sulfur, salicylaldehyde (10), aryldiazonium chlorides 12a,b and ω-bromo-4-methoxy- acetophenone (14), which afforded, fused thiophene derivatives 9a,b, coumarin derivative 11, arylhdrazono derivatives 13a,b and 4-methoxyphenyl butenyl derivative 15, respectively. The latter product 15 was reacted with either potassium cyanide (16a) or potassium thiocyanide (16b) to form cyano and thiocyano derivatives 17a,b, respectively. Compound 17a underwent further cyclization to afford pyridopyrimidine derivative 19. Compound 15 was reacted with either hydrazine (20a) or phenylhydrazine (20b) to produce hydrazo derivatives 21a,b and these products were cyclize to produce pyrrole derivatives 23a,b. Finally, 5-ureidohydantion (2) was reacted with compounds 24a,b,c to afford pyrimidine derivatives 25a,b,c. The structures of the synthesized compounds were confirmed using IR, 1 H NMR, 13C NMR and mass spectrometry techniques. Compounds 11 and 19 have promising as analgesic and antipyretic activities

The Benign Prostatic Hyperplasia and Its Aetiologies

This study aimed at investigating the Benign Prostatic Hyperplasia and Its Aetiologies, therefore th prostatic hyperplasia predominantly involves the stromal compartment of the gland and affects more than 70% of men of 70 years or older with or without obstructive symptoms of benign prostatic hyperplasia. A consensus view is emerging concerning the factors and control systems that modulate cell proliferation and connective tissue biology in the prostate. The purpose of this review is to discuss some of the recent work contributing to the latter in the context of the aetiology of benign prostatic hyperplasia. The current study also reviews the most important findings regarding the key mechanisms involved in the pathophysiology of BPH. The study concluded that although the pathogenesis of BPH is not yet fully understood, several mechanisms seem to be involved in the development and progression of the disease. These mainly include systemic and local hormonal and vascular alterations as well as prostatic inflammation that would stimulate cellular proliferation

Gestational diabetes prevalence and risk factors among pregnant women — Jazan Region, Saudi Arabia

Background. The gestational diabetes (GDM) is considered as a global public health problem that affects pregnant women. GDM can become chronic type II and usually it is associated with many risk factors that may lead to many serious complications for the mother and the fetus. The main objective of this study was to estimate the prevalence of GDM among pregnant women in Jazan region and to determine the possible associated factors of the GDM. Material and methods. The study involved pregnant women attending department of Obstetrics and Gynecology at government hospitals of Jazan region. A sample of 440 pregnant women were randomly selected. Interviews were conducted using a questionnaire prepared to measure the GDM prevalence, risk factors, awareness and adherence to the advice of the doctor and medications. Results. The prevalence of gestational diabetes among pregnant women in Jazan is estimated at 8.2%. The GDM prevalence was significantly higher among obese women (20.2%; 95% CI 13.2–29.2) compared with women with normal weight (7.1%; 95% CI 1.7–7.6). The analysis showed that GDM was significantly associated with child weight more than 3.5 kg (OR 4.315; p = 0.004), mother’s BMI more than 30 kg/m2 (OR 4.703; p = 0.001), and family history of GDM (OR 2.606; p = 0.046). Conclusion. In conclusion, the GDM prevalence obtained in this study is more than global prevalence and less than other studies in KSA. The BMI of mothers and having neonates that weight more than 3.5 kg are the main risk factors for GDM. Suitable interventions programs are highly required for control and risk factor modifications

Overview On Surgical Management Of Overactive Bladder

Overactive bladder syndrome is a persistent and incapacitating disorder that has profound medical, psychological, and social implications, greatly impacting the wellbeing of countless individuals globally. A significant number of individuals experience urine urgency, which can be extremely bothersome. The primary indicator of overactive bladder (OAB) is a sense of urgency, often accompanied by increased urine frequency and nocturia. After ruling out other medical conditions with similar symptoms, the initial approach to managing OAB is providing guidance on fluid consumption and bladder training. If needed, antimuscarinic medicines may be added as a supplement. If patients have significant distress from OAB symptoms even after maximizing medicinal treatment, they may choose to undergo invasive procedures. There is currently a limited understanding of the hierarchical structure of central nervous system control. However, the use of functional imaging is starting to reveal the difficulties that need to be addressed in this area. Current research is exploring the use of botulinum neurotoxin-A injection, oral β3-adrenergic agonists, and innovative methods for nerve stimulation as potential therapies. The inherent subjectivity of urine urgency, the absence of animal models, and the complex pathophysiology of overactive bladder (OAB) pose substantial obstacles to achieving effective clinical therapy

Integrating network pharmacology with molecular docking to rationalize the ethnomedicinal use of Alchornea laxiflora (Benth.) Pax & K. Hoffm. for efficient treatment of depression

Background: Alchornea laxiflora (Benth.) Pax & K. Hoffm. (A. laxiflora) has been indicated in traditional medicine to treat depression. However, scientific rationalization is still lacking. Hence, this study aimed to investigate the antidepressant potential of A. laxiflora using network pharmacology and molecular docking analysis.Materials and methods: The active compounds and potential targets of A. laxiflora and depression-related targets were retrieved from public databases, such as PubMed, PubChem, DisGeNET, GeneCards, OMIM, SwissTargetprediction, BindingDB, STRING, and DAVID. Essential bioactive compounds, potential targets, and signaling pathways were predicted using in silico analysis, including BA-TAR, PPI, BA-TAR-PATH network construction, and GO and KEGG pathway enrichment analysis. Later on, with molecular docking analysis, the interaction of essential bioactive compounds of A. laxiflora and predicted core targets of depression were verified.Results: The network pharmacology approach identified 15 active compounds, a total of 219 compound-related targets, and 14,574 depression-related targets with 200 intersecting targets between them. SRC, EGFR, PIK3R1, AKT1, and MAPK1 were the core targets, whereas 3-acetyloleanolic acid and 3-acetylursolic acid were the most active compounds of A. laxiflora with anti-depressant potential. GO functional enrichment analysis revealed 129 GO terms, including 82 biological processes, 14 cellular components, and 34 molecular function terms. KEGG pathway enrichment analysis yielded significantly enriched 108 signaling pathways. Out of them, PI3K-Akt and MAPK signaling pathways might have a key role in treating depression. Molecular docking analysis results exhibited that core targets of depression, such as SRC, EGFR, PIK3R1, AKT1, and MAPK1, bind stably with the analyzed bioactive compounds of A. laxiflora.Conclusion: The present study elucidates the bioactive compounds, potential targets, and pertinent mechanism of action of A. laxiflora in treating depression. A. laxiflora might exert an antidepressant effect by regulating PI3K-Akt and MAPK signaling pathways. However, further investigations are required to validate

Częstość występowania cukrzycy ciążowej i jej czynników ryzyka u ciężarnych kobiet w prowincji Dżazan w Arabii Saudyjskiej

Wstęp. Cukrzycę ciążową (GDM) uważa się za globalny problem zdrowotny dotyczący ciężarnych kobiet. Może ona przejść w cukrzycę typu 2 i zwykle wiąże się z wieloma czynnikami ryzyka, które mogą prowadzić do poważnych powikłań u matki i płodu. Głównym celem badania było oszacowanie częstości GDM u kobiet w ciąży mieszkających w prowincji Dżazan oraz określenie czynników, które mogą wiązać się z GDM. Materiał i metody. Badanie obejmowało kobiety w ciąży będące pod opieką oddziałów położniczo-ginekologicznych szpitali państwowych w prowincji Dżazan. Wybrano losowo próbę liczącą 440 ciężarnych kobiet. Przeprowadzając wywiady z uczestniczkami badania, korzystano z kwestionariusza opracowanego w celu oceny częstości występowania GDM, czynników ryzyka, wiedzy kobiet na temat GDM i przestrzegania zaleceń lekarskich oraz przyjmowania leków. Wyniki. Częstość GDM wśród ciężarnych kobiet w prowincji Dżazan wynosiła 8,2%. Była ona istotnie wyższa u kobiet otyłych [20,2%; 95-procentowy przedział ufności (CI) 13,2–29,2) niż u osób z prawidłową masą ciała (7,1%; 95% CI 1,7–7,6). Analiza wykazała, że GDM wiązała się istotnie z urodzeniem dziecka o masie ciała wynoszącej ponad 3,5 kg [iloraz szans (OR) 4,315; p = 0,004), wskaźnikiem masy ciała (BMI) matki wynoszącym ponad 30 kg/m2 (OR 4,703; p = 0,001) oraz dodatnim wywiadem rodzinnym dotyczącym GDM (OR 2,606; p = 0,046). Wnioski. Podsumowując, częstość GDM stwierdzona w badaniu była większa niż globalna częstość występowania GDM, lecz mniejsza niż obserwowana w innych badaniach w populacji saudyjskiej. Wskaźnik BMI matki i urodzenie dziecka o masie ciała powyżej 3,5 kg to główne czynniki ryzyka GDM. Potrzebne są odpowiednie programy interwencyjne w celu kontroli i modyfikacji czynników ryzyka

Antimicrobial resistance among migrants in Europe: a systematic review and meta-analysis

BACKGROUND: Rates of antimicrobial resistance (AMR) are rising globally and there is concern that increased migration is contributing to the burden of antibiotic resistance in Europe. However, the effect of migration on the burden of AMR in Europe has not yet been comprehensively examined. Therefore, we did a systematic review and meta-analysis to identify and synthesise data for AMR carriage or infection in migrants to Europe to examine differences in patterns of AMR across migrant groups and in different settings. METHODS: For this systematic review and meta-analysis, we searched MEDLINE, Embase, PubMed, and Scopus with no language restrictions from Jan 1, 2000, to Jan 18, 2017, for primary data from observational studies reporting antibacterial resistance in common bacterial pathogens among migrants to 21 European Union-15 and European Economic Area countries. To be eligible for inclusion, studies had to report data on carriage or infection with laboratory-confirmed antibiotic-resistant organisms in migrant populations. We extracted data from eligible studies and assessed quality using piloted, standardised forms. We did not examine drug resistance in tuberculosis and excluded articles solely reporting on this parameter. We also excluded articles in which migrant status was determined by ethnicity, country of birth of participants' parents, or was not defined, and articles in which data were not disaggregated by migrant status. Outcomes were carriage of or infection with antibiotic-resistant organisms. We used random-effects models to calculate the pooled prevalence of each outcome. The study protocol is registered with PROSPERO, number CRD42016043681. FINDINGS: We identified 2274 articles, of which 23 observational studies reporting on antibiotic resistance in 2319 migrants were included. The pooled prevalence of any AMR carriage or AMR infection in migrants was 25·4% (95% CI 19·1-31·8; I2 =98%), including meticillin-resistant Staphylococcus aureus (7·8%, 4·8-10·7; I2 =92%) and antibiotic-resistant Gram-negative bacteria (27·2%, 17·6-36·8; I2 =94%). The pooled prevalence of any AMR carriage or infection was higher in refugees and asylum seekers (33·0%, 18·3-47·6; I2 =98%) than in other migrant groups (6·6%, 1·8-11·3; I2 =92%). The pooled prevalence of antibiotic-resistant organisms was slightly higher in high-migrant community settings (33·1%, 11·1-55·1; I2 =96%) than in migrants in hospitals (24·3%, 16·1-32·6; I2 =98%). We did not find evidence of high rates of transmission of AMR from migrant to host populations. INTERPRETATION: Migrants are exposed to conditions favouring the emergence of drug resistance during transit and in host countries in Europe. Increased antibiotic resistance among refugees and asylum seekers and in high-migrant community settings (such as refugee camps and detention facilities) highlights the need for improved living conditions, access to health care, and initiatives to facilitate detection of and appropriate high-quality treatment for antibiotic-resistant infections during transit and in host countries. Protocols for the prevention and control of infection and for antibiotic surveillance need to be integrated in all aspects of health care, which should be accessible for all migrant groups, and should target determinants of AMR before, during, and after migration. FUNDING: UK National Institute for Health Research Imperial Biomedical Research Centre, Imperial College Healthcare Charity, the Wellcome Trust, and UK National Institute for Health Research Health Protection Research Unit in Healthcare-associated Infections and Antimictobial Resistance at Imperial College London

Burnout among surgeons before and during the SARS-CoV-2 pandemic: an international survey

Background: SARS-CoV-2 pandemic has had many significant impacts within the surgical realm, and surgeons have been obligated to reconsider almost every aspect of daily clinical practice. Methods: This is a cross-sectional study reported in compliance with the CHERRIES guidelines and conducted through an online platform from June 14th to July 15th, 2020. The primary outcome was the burden of burnout during the pandemic indicated by the validated Shirom-Melamed Burnout Measure. Results: Nine hundred fifty-four surgeons completed the survey. The median length of practice was 10 years; 78.2% included were male with a median age of 37 years old, 39.5% were consultants, 68.9% were general surgeons, and 55.7% were affiliated with an academic institution. Overall, there was a significant increase in the mean burnout score during the pandemic; longer years of practice and older age were significantly associated with less burnout. There were significant reductions in the median number of outpatient visits, operated cases, on-call hours, emergency visits, and research work, so, 48.2% of respondents felt that the training resources were insufficient. The majority (81.3%) of respondents reported that their hospitals were included in the management of COVID-19, 66.5% felt their roles had been minimized; 41% were asked to assist in non-surgical medical practices, and 37.6% of respondents were included in COVID-19 management. Conclusions: There was a significant burnout among trainees. Almost all aspects of clinical and research activities were affected with a significant reduction in the volume of research, outpatient clinic visits, surgical procedures, on-call hours, and emergency cases hindering the training. Trial registration: The study was registered on clicaltrials.gov "NCT04433286" on 16/06/2020

Surgical site infection after gastrointestinal surgery in high-income, middle-income, and low-income countries: a prospective, international, multicentre cohort study

Background: Surgical site infection (SSI) is one of the most common infections associated with health care, but its importance as a global health priority is not fully understood. We quantified the burden of SSI after gastrointestinal surgery in countries in all parts of the world. Methods: This international, prospective, multicentre cohort study included consecutive patients undergoing elective or emergency gastrointestinal resection within 2-week time periods at any health-care facility in any country. Countries with participating centres were stratified into high-income, middle-income, and low-income groups according to the UN's Human Development Index (HDI). Data variables from the GlobalSurg 1 study and other studies that have been found to affect the likelihood of SSI were entered into risk adjustment models. The primary outcome measure was the 30-day SSI incidence (defined by US Centers for Disease Control and Prevention criteria for superficial and deep incisional SSI). Relationships with explanatory variables were examined using Bayesian multilevel logistic regression models. This trial is registered with ClinicalTrials.gov, number NCT02662231. Findings: Between Jan 4, 2016, and July 31, 2016, 13 265 records were submitted for analysis. 12 539 patients from 343 hospitals in 66 countries were included. 7339 (58·5%) patient were from high-HDI countries (193 hospitals in 30 countries), 3918 (31·2%) patients were from middle-HDI countries (82 hospitals in 18 countries), and 1282 (10·2%) patients were from low-HDI countries (68 hospitals in 18 countries). In total, 1538 (12·3%) patients had SSI within 30 days of surgery. The incidence of SSI varied between countries with high (691 [9·4%] of 7339 patients), middle (549 [14·0%] of 3918 patients), and low (298 [23·2%] of 1282) HDI (p < 0·001). The highest SSI incidence in each HDI group was after dirty surgery (102 [17·8%] of 574 patients in high-HDI countries; 74 [31·4%] of 236 patients in middle-HDI countries; 72 [39·8%] of 181 patients in low-HDI countries). Following risk factor adjustment, patients in low-HDI countries were at greatest risk of SSI (adjusted odds ratio 1·60, 95% credible interval 1·05–2·37; p=0·030). 132 (21·6%) of 610 patients with an SSI and a microbiology culture result had an infection that was resistant to the prophylactic antibiotic used. Resistant infections were detected in 49 (16·6%) of 295 patients in high-HDI countries, in 37 (19·8%) of 187 patients in middle-HDI countries, and in 46 (35·9%) of 128 patients in low-HDI countries (p < 0·001). Interpretation: Countries with a low HDI carry a disproportionately greater burden of SSI than countries with a middle or high HDI and might have higher rates of antibiotic resistance. In view of WHO recommendations on SSI prevention that highlight the absence of high-quality interventional research, urgent, pragmatic, randomised trials based in LMICs are needed to assess measures aiming to reduce this preventable complication