Combined Gastric and Colorectal Cancer Screening—A New Strategy

Background: Our aim was to evaluate the feasibility of a serological assessment of gastric cancer risk in patients undergoing colonoscopy in countries with low-to-moderate incidence rates. Methods: Serum samples were prospectively collected from 453 patients (>50 years old) undergoing colonoscopies. Of these, 279 (61.6%) also underwent gastroscopy to correlate the results for serum pepsinogen I and II (sPG-I and sPG-II), sPG-I/II ratio, and anti-H. pylori antibodies with gastric histopathology findings (graded according to the updated Sydney classification and the Operative Link of Gastritis Assessment (OLGA) and the Operative Link for Gastric Intestinal Metaplasia assessment (OLGIM) systems). Results: H. pylori was found in 85 patients (30.5%). Chronic atrophic gastritis was diagnosed in 89 (31.9%) patients. High-risk OLGA (III-IV) stages were present in 24 patients, and high-risk OLGIM stages were present in 14 patients. There was an inverse correlation of sPG-I with the degree of atrophy and intestinal metaplasia (IM), as well as with the respective OLGA (r = -0.425; p < 0.001) and OLGIM (r = -0.303; p < 0.001) stages. A pathological sPG-I result was associated with a relative risk (RR) of 12.2 (95% confidence interval: 6.29-23.54; p < 0.001) for gastric preneoplastic changes. Conclusions: The assessment of serum pepsinogen allows the identification of patients at increased risk of gastric cancer. A prevention strategy of combining a screening colonoscopy with a serological screening for preneoplastic gastric changes should be considered in the general population

Hemijski sastav i antioksidantna aktivnost metanolnog ekstrakta kore stabla Sorbus torminalis

Sorbus torminalis (L.) Crantz (Rosaceae), also known as wild service tree, is a deciduous tree widespread in central and southern Europe, north-western Africa, western Asia, the Balkan Peninsula and the Caucasus (1, 2). Species of the genus Sorbus represent a rich source of polyphenols that are responsible for their numerous proven effects, including anti-diarrhoeal, anti-inflammatory, antidiabetic, antioxidant and anti-atherogenic effect. Accordingly, the bark of S. torminalis could have interesting composition and effects, however, data about this herbal drug is insufficient (1). Bark dry methanol extract (DER 9.74:1), obtained after successive extraction with cyclohexane and dichloromethane, was analyzed for antioxidant potential using DPPH and FRAP assay. Contents of total polyphenols, flavonoids and tannins were determined using spectrophotometric methods based on reaction with Folin-Ciocalteu reagent (FC), reaction with aluminum-chloride and adsorption by hide powder in combination with reaction with FC, respectively. LC-DAD-ESI- MS analysis was used for identification of main extract constituents. In DPPH test IC50 value 8.66±0.07 μg/mL was shown, while result of FRAP test was 3.01±0.11 μmol Fe 2+ /mg of dry extract. Contents of total polyphenols, flavonoids and tannins were 0.39±0.06 mg GA/mg of dry extract, 0.035±0.019% and 1.3±0.08%, respectively. By LC-DAD-ESI-MS analysis 25 compounds were detected among which procyanidins (dimers, trimers and tetramers) are dominant. The results of this study showed that dry methanol extract of S. torminalis bark is rich in procyanidins which potentially contribute to its antioxidant activity. Additional significant effects of this herbal drug and its isolated compounds should be investigated in future studies.Sorbus torminalis (L.) Crantz (Rosaceae) je listopadno drvo, u narodu poznato pod nazivom brekinja. Rasprostranjeno je širom centralne i južne Evrope, severozapadne Afrike, zapadne Azije, Balkanskog poluostrva i Kavkaza (1, 2). Vrste roda Sorbus predstavljaju bogat izvor polifenola koji su odgovorni za njihove brojne dokazane efekte, uključuju antidijaroični, antiinflamatorni, antidijabetski, antioksidantni i antiaterogeni efekat. U skladu sa tim, kora S. torminalis mogla bi imati interesantan sastav i efekte, međutim, o ovoj biljnoj drogi nema dovoljno podataka (1). Antioksidantna aktivnost suvog metanolnog ekstrakta kore stabla (DER 9,74:1), dobijenog nakon sukcesivne ekstrakcije cikloheksanom i dihlormetanom, analizirana je DPPH i FRAP testom. Sadržaji ukupnih polifenola, flavonoida i tanina određeni su spektrofotometrijskim metodama koje se zasnivaju na reakciji sa Folin‐Ciocalteu reagensom (FC), reakciji sa aluminijum-hloridom i na adsorpciji na kožni prah u kombinaciji sa reakcijom sa FC reagensom, redom. LC‐DAD‐ESI‐MS analiza korišćena je za identifikaciju glavnih sastojaka ekstrakta. U DPPH testu dobijena je IC50 vrednost 8,66±0,07 μg/mL, a rezultat FRAP testa bio je 3,01±0,11 μmol Fe 2+ /mg suvog ekstrakta. Sadžaj ukupnih polifenola bio je 0,39±0,06 mg GA/mg suvog ekstrakta, flavonoida 0,035±0,019%, a tanina 1,3±0,08%. LC‐DAD‐ESI‐MS analizom detektovano je 25 jedinjenja od kojih su dominantni procijanidini (dimeri, trimeri i tetrameri). Rezultati ovog istraživanja pokazali su da je suvi metanolni ekstrakt kore stabla S. torminalis bogat procijanidinima koji potencijalno doprinose njegovoj antioksidantnoj aktivnosti. Dodatne značajne efekte ove biljne droge i njenih izolovanih jedinjenja trebalo bi ispitati u budućim istraživanjima.VIII Kongres farmaceuta Srbije sa međunarodnim učešćem, 12-15.10.2022. Beogra

Stabilization of Black Locust Flower Extract via Encapsulation Using Alginate and Alginate–Chitosan Microparticles

Black locust flower extract contains various polyphenols and their glucosides contribute to the potential health benefits. After intake of these bioactive compounds and passage through the gastrointestinal tract, their degradation can occur and lead to a loss of biological activity. To overcome this problem, the bioactive compounds should be protected from environmental conditions. This study aimed to encapsulate the black flower extract in the microparticles based on biodegradable polysaccharides, alginate, and chitosan. In the extract, the total antioxidant content was found to be 3.18 ± 0.01 g gallic acid equivalent per 100 g of dry weight. Also, the presence of lipids (16), phenolics (27), organic acids (4), L-aspartic acid derivative, questinol, gibberellic acid, sterol, and saponins (2) was confirmed using the UHPLC–ESI–MS analysis. In vitro assays showed that the extract has weak anti-α-glucosidase activity and moderate antioxidant and cytotoxic activity against the HeLa cell line. The extrusion method with secondary air flow enabled the preparation of microparticles (about 270 μm) encapsulated with extract. An encapsulation efficiency of over 92% was achieved in the alginate and alginate–chitosan microparticles. The swelling study confirmed a lower permeability of alginate–chitosan microparticles compared with alginate microparticles. For both types of microparticles, the release profile of antioxidants in the simulated gastrointestinal fluids at 37 °C followed the Korsmeyer–Peppas model. A lower diffusion coefficient than 0.5 indicated the simple Fick diffusion of antioxidants. The alginate–chitosan microparticles enabled a more sustained release of antioxidants from extract compared to the alginate microparticles. The obtained results indicated an improvement in the antioxidant activity of bioactive compounds from the extract and their protection from degradation in the simulated gastric conditions via encapsulation in the polymer matrixes. Alginate–chitosan showed slightly slower cumulative antioxidant release from microparticles and better antioxidant activity of the extract compared to the alginate system. According to these results, alginate–chitosan microparticles are more suitable for further application in the encapsulation of black locust flower extract. Also, the proposed polymer matrix as a drug delivery system is safe for human use due to its biodegradability and non-toxicity

Antiproliferative activity and mode of action analysis of novel amino and amido substituted phenantrene and naphtho[2,1-b]thiophene derivatives

Herein we present and describe the design and synthesis of novel phenantrene derivatives substituted with either amino or amido side chains and their biological activity. Antiproliferative activities were assessed in vitro on a panel of human cancer cell lines. Tested compounds showed moderate activity against cancer cells in comparison with 5-fluorouracile. Among all tested compounds, some compounds substituted with cyano groups showed a pronounced and selective activity in the nanomolar range of inhibitory concentrations against HeLa and HepG2. The strongest selective activity against HeLa cells was observed for acrylonitriles 8 and 11 and their cyclic analogues 15 and 17 substituted with two cyano groups with a corresponding IC50 = 0.33, 0.21, 0.65 and 0.45 μM, respectively. Compounds 11 showed the most pronounced selectivity being almost non cytotoxic to normal fibroblasts. Additionally, mode of biological action analysis was performed in silico and in vitro by Western blot analysis of HIF-1-α relative expression for compounds 8 and 11

Inhibicija Α‐amilaze in vitro i in vivo hipoglikemijski efekat metanolnog ekstrakta herbe Alchemilla viridiflora Rothm

Dry methanol extract (DER 3.95:1) of aerial flowering parts of Alchemilla viridiflora Rothm., Rosaceae, obtained after successive extraction with cyclohexane and dichloromethane, predominantly contains polyphenols, tannins (3.74%), mostly ellagic, and flavonoids (0.30%), hexosides of quercetin and kaempferol. It shows ACE inhibitory and anti-Helicobacter pylori activity in vitro (1). Based on the traditional use of the common A. vulgaris in the treatment of diabetes (2), the effect of this A. viridiflora extract on α-amylase activity in vitro and lowering blood glucose in rats were investigated. α-Amylase inhibitory activity was assessed using the 3,5-dinitrosalicylic acid method. Extract in doses of 50, 100 and 200 mg/kg, p.o., was administered to streptozotocin-induced diabetic rats for 20 days, and blood glucose level and weight of rats were monitored during treatment. Extract inhibited α-amylase activity at IC 50 5.47 ± 0.30 mg/mL. A dose of 200 mg/kg significantly decreased blood glucose level after 10 (32.2%) and 20 days of treatment (38.3%). The effect of this dose was similar to the effect of the reference drug, glibenclamide. A dose of 100 mg/kg significantly increased glucose level by day 10 (50.5%) and a reduction in blood sugar level of 51.2% was observed only after 10 days. Body weight was correlated with changes in blood sugar levels. A dose of 50 mg/kg had no statistically significant effect. Results indicate a significant potential of A. viridiflora aerial parts in the treatment of diabetes, which must be confirmed by additional studies, and active ingredients need to be identified.Suvi metanolni ekstrakt (DER 3,95:1) nadzemnih delova u cvetu Alchemilla viridiflora Rothm., Rosaceae, dobijen nakon sukcesivne ekstrakcije cikloheksanom i dihlormetanom, sadrži visok nivo polifenola, tanina (3,74%), uglavnom elagnih, i flavonoida (0,30%), heksozida kvercetina i kemferola. Pokazuje ACE inhibitornu i anti-Helicobacter pylori aktivnost in vitro (1). Na osnovu podataka o tradicionalnoj upotrebi herbe A. vulgaris, najčešće korišćene Alchemilla vrste, u lečenju dijabetesa (2), ispitivan je uticaj navedenog ekstrakta A. viridiflora na aktivnost α-amilaze in vitro i na snižavanje nivoa glukoze u krvi kod pacova. Uticaj na aktivnost α-amilaze procenjen je primenom metode sa 3,5- dinitrosalicilnom kiselinom. Ekstrakt A. viridiflora u dozama od 50, 100 i 200 mg/kg, p.o., aplikovan je pacovima sa dijabetesom izazvanim streptozotocinom tokom 20 dana, a nivoi glukoze u krvi i težina pacova su praćeni u određenim vremenskim intervalima tokom trajanja tretmana. Ekstrakt je inhibirao aktivnost α-amilaze, a vrednost IC 50 iznosila je 5,47±0,30 mg/mL. Doza od 200 mg/kg značajno je smanjila nivo glukoze u krvi nakon 10 (32,2%) i 20 dana tretmana (38,3%). Efekat ove doze bio je sličan efektu referentnog leka, glibenklamida. S druge strane, doza od 100 mg/kg značajno je povećala nivo glukoze do 10. dana (50,5%) a do značajnog poboljšanja, tj. sniženja nivoa šećera od 51,2% dovela je tek nakon 10 dana. Telesna težina bila je u korelaciji sa promenama nivoa šećera u krvi. Doza od 50 mg/kg nije imala statistički značajan efekat. Rezultati ukazuju na značajan potencijal herbe A. viridiflora u lečenju dijabetesa, što mora biti potvrđeno dodatnim studijama, a aktivni sastojci identifikovani.VIII Kongres farmaceuta Srbije sa međunarodnim učešćem, 12-15.10.2022. Beogra

The influence of gastric atrophy on Helicobacter pylori antibiotics resistance in therapy-naïve patients

Background: Antibiotic susceptibility of Helicobacter pylori to antibiotics may vary among different niches of the stomach. The progression of chronic H. pylori gastritis to atrophy changes intragastric physiology that may influence selection of resistant strains. Aim: To study the antibiotic resistance of H. pylori taking the severity of atrophic gastritis in antrum and corpus into account. Methods: Helicobacter pylori-positive patients (n = 110, m = 32, mean age 52.6 ± 13.9 years) without prior H. pylori eradication undergoing upper gastrointestinal (GI) endoscopy for dyspeptic symptoms were included in a prospective study. Patients were stratified into three groups depending on the grade of atrophy: no atrophy (OLGA Stage 0), mild atrophy (OLGA Stage I–II) and moderate/severe atrophy (OLGA Stage III–IV). Two biopsies each from the antrum and the corpus and one from the angulus were taken and assessed according to the updated Sydney system. H. pylori strains were isolated from antrum and corpus biopsies and tested for antibiotic susceptibility (AST) for amoxicillin, clarithromycin, metronidazole, levofloxacin, tetracycline, and rifampicin by the agar dilution methods. A Chi-square test of independence with a 95% confidence interval was used to detect differences in the proportion of patients with susceptible and resistant H. pylori strains. Results: Among 110 patients, primary clarithromycin resistance (R) was 30.0%, both in the antrum and corpus; metronidazole resistance accounted for 36.4 and 34.5% in the antrum and corpus; and levofloxacin was 19.1 and 22.7% in the antrum and corpus, respectively. Resistance rates to amoxicillin, tetracycline, and rifampicin were below 5%. Dual antibiotic resistance rate was 21.8%, and triple resistance rate was 9.1%. There was a significant difference in the resistance rate distribution in antrum (p < 0.0001) and corpus (p < 0.0001). With increasing severity of atrophy according to OLGA stages, there was a significant increase in clarithromycin-R and metronidazole-R. Conclusion: In treatment-naïve patients, antibiotic resistance and heteroresistance were related to the severity of atrophy. The high clarithromycin resistance in atrophic gastritis suggests that H. pylori antibiotic susceptibility testing should always be performed in this condition before selecting the eradication regimen

Epigenetic regulation of Delta-Like1 controls Notch1 activation in gastric cancer

The Notch signaling pathway drives proliferation, differentiation, apoptosis, cell fate, and maintenance of stem cells in several tissues. Aberrant activation of Notch signaling has been described in several tumours and in gastric cancer (GC), activated Notch1 has been associated with de-differentiation of lineage-committed stomach cells into stem progenitors and GC progression. However, the specific role of the Notch1 ligand (DLL1) in GC has not yet been elucidated. To assess the role of DLL1 in GC cancer, the expression of Notch1 and its ligands DLL1 and Jagged1, was analyzed in 8 gastric cancer cell lines (KATOIII, SNU601, SNU719, AGS, SNU16, MKN1, MKN45, TMK1). DLL1 expression was absent in KATOIII, SNU601, SNU719 and AGS. The lack of DLL1 expression in these cells was associated with promoter hypermethylation and 5-aza-2’deoxycitidine caused up-regulation of DLL1. The increase in DLL1 expression was associated with activation of Notch1 signalling, with an increase in cleaved Notch1 intracellular domain (NICD) and Hes1, and down-regulation in Hath1. Concordantly, Notch1 signalling was activated with the overexpression of DLL1. Moreover, Notch1 signalling together with DLL1 methylation were evaluated in samples from 52 GC patients and 21 healthy control as well as in INS-GAS mice infected with H. pylori and randomly treated with eradication therapy. In GC patients, we found a correlation between DLL1 and Hes1 expression, while DLL1 methylation and Hath1 expression were associated with the diffuse and mixed type of gastric cancer. Finally, none of the samples from INS-GAS mice infected with H. pylori, a model of intestinal-type gastric tumorigenesis, showed promoter methylation of DLL1. This study shows that Notch1 activity in gastric cancer is controlled by the epigenetic silencing of the ligand DLL1, and that Notch1 inhibition is associated with the diffuse type of gastric cancer

In Silico and In Vitro Studies of Alchemilla viridiflora Rothm-Polyphenols' Potential for Inhibition of SARS-CoV-2 Internalization

Since the outbreak of the COVID-19 pandemic, it has been obvious that virus infection poses a serious threat to human health on a global scale. Certain plants, particularly those rich in polyphe- nols, have been found to be effective antiviral agents. The effectiveness of Alchemilla viridiflora Rothm. (Rosaceae) methanol extract to prevent contact between virus spike (S)-glycoprotein and angiotensin- converting enzyme 2 (ACE2) and neuropilin-1 (NRP1) receptors was investigated. In vitro results revealed that the tested samples inhibited 50% of virus-receptor binding interactions in doses of 0.18 and 0.22 mg/mL for NRP1 and ACE2, respectively. Molecular docking studies revealed that the compounds from A. viridiflora ellagitannins class had a higher affinity for binding with S-glycoprotein whilst flavonoid compounds more significantly interacted with the NRP1 receptor. Quercetin 3-(6”-ferulylglucoside) and pentagalloylglucose were two compounds with the highest exhibited interfering potential for selected target receptors, with binding energies of −8.035 (S-glycoprotein) and −7.685 kcal/mol (NRP1), respectively. Furthermore, computational studies on other SARS-CoV-2 strains resulting from mutations in the original wild strain (V483A, N501Y-K417N-E484K, N501Y, N439K, L452R-T478K, K417N, G476S, F456L, E484K) revealed that virus internalization activity was maintained, but with different single compound contributions

Hepatobiliary MRI: Signal intensity based assessment of liver function correlated to 13C-Methacetin breath test

Gadoxetic acid (Gd-EOB-DTPA) is a paramagnetic MRI contrast agent with raising popularity and has been used for evaluation of imaging-based liver function in recent years. In order to verify whether liver function as determined by real-time breath analysis using the intravenous administration of C-13-methacetin can be estimated quantitatively from Gd-EOB-DTPA-enhanced MRI using signal intensity (SI) values. 110 patients underwent Gd-EOB-DTPA-enhanced 3-T MRI and, for the evaluation of liver function, a C-13-methacetin breath test (C-13-MBT). SI values from before (SIpre) and 20 min after (SIpost) contrast media injection were acquired by T1-weighted volume-interpolated breath-hold examination (VIBE) sequences with fat suppression. The relative enhancement (RE) between the plain and contrast-enhanced SI values was calculated and evaluated in a correlation analysis of C-13-MBT values to SIpost and RE to obtain a SI-based estimation of C-13-MBT values. The simple regression model showed a log-linear correlation of C-13-MBT values with SIpost and RE (p < 0.001). Stratified by 3 different categories of C-13-MBT readouts, there was a constant significant decrease in both SIpost (p <= 0.002) and RE (p <= 0.033) with increasing liver disease progression as assessed by the C-13-MBT. Liver function as determined using real-time C-13-methacetin breath analysis can be estimated quantitatively from GdEOB- DTPA-enhanced MRI using SI-based indices