Search

Antimicrobial resistance (AMR) is an ongoing phenomenon. It is a significant public health issue that has existed long before the coronavirus disease of 2019 (COVID-19) pandemic. It develops as microorganisms undergo genetic mutations that allow them to survive despite antimicrobial treatment. This process is highly associated with excessive and often unnecessary antimicrobial pharmacotherapy, which was often discussed during the COVID-19 pandemic. This article explores how the pandemic has affected antimicrobial stewardship by shifting the focus away from antimicrobial resistance, as well as the impact of enhanced antibiotic usage and measures such as lockdowns, mandatory testing and vaccination on antimicrobial resistance. Although these measures were regarded as successful in terms of limiting the pandemic, they have significantly contributed to an already escalating AMR issue. Outpatient methods in primary care and intensive care units aiming to prevent severe COVID-19 disease have contributed to the spread of multidrug-resistant bacteria, while laboratories burdened with COVID- 19 testing have indirectly interrupted the detection of these bacteria. In this review, we summarize the pathogens whose AMRe has been greatly affected by COVID-19 measures and emphasize the importance of efficient antimicrobial stewardship in future pandemic and non-pandemic states to promote the responsible use of antibiotics and minimize AMR.

Background: With the increasing prevalence of obesity, fatty liver injury, and drug use, appropriate treatment of non-alcoholic (NAFLD) and drug-induced fatty liver disease (DIFLD) is becoming more important. Numerous studies have proposed and tested a number of potential hepatoprotective agents. The aim of this study was to establish reliable cell culture models for NAFLD and DIFLD and to determine the effect of the GLP-1 agonist liraglutide in these models. Methods: Huh7 cells were grown overnight to reach 90% confluency, followed by 24h of incubation according to further described protocols. Experiments were performed in triplicates, with the following cell subgroups: Huh7 cells incubated in medium only as a negative control, cells incubated with increasing concentrations of liraglutide (5nM, 10nM, 20nM), cells treated with 0, 5 mM oleic acid (OA) (NAFLD model) , and/or cotreated with liraglutide, and cells treated with 20μM amiodarone (DIFLD model), and/or cotreated with liraglutide. The antiproliferative and cytotoxic effects were determined using a colorimetric MTT assay. Cells were stained with Oil-Red-O dye and Fluorescent mounting medium with DAPI, and visualized under a microscope. ImageJ software was used to count the cell nuclei and measure the integrated density relative to the cell count. After extraction of total RNA with NucleoZol, cDNA synthesis, PCR method and gel electrophoresis were performed, and finally the signals were analyzed by ImageJ software. Results: Both oleic acid and amiodarone reduced cell viability by 30% (p

Background and aims: The impact of non-alcoholic fatty liver disease (NAFLD) on global health is becoming more significant with the growing incidence of obesity and drugs consumption. Therefore, more research is necessary in order to enlighten underlying pathophysiologic mechanisms for these conditions and possible therapeutic approaches. Aims of our study were to establish reliable in vitro models of non-alcoholic and drug-induced fatty liver disease (DIFLD) and also to investigate possible therapeutic solutions in these in vitro models. Method: HuH7 cell culture models of NAFLD and DIFLD were established by incubation of Huh7 cells with 0, 5 mM oleic acid (OA), 5 μM to 20 μM of amiodarone, 1 μM to 10 μM of tamoxifen for various time periods (24 h, 48 h), respectively. Cells where cotreated with 1 nM to 100 nM of liraglutide in order to assess its potential beneficial effect in fatty liver models. Cell viability was measured by MTT ((3- (4, 5-dimethylthiazol-2-yl)-2, 5- diphenyltetrazolium bromide) assay. Changes in cell shape and the extent of hepatosteatosis were assessed by fluorescent microscopy as shown in Figure. Cell nuclei were stained with Hoechst, and fat droplets were stained with Oil-Red-O. Trigliceryde (TG) accumulation was measured by TGO-PAP method and the results were read on the microplate reader. Results: Tamoxifen significantly reduced cell viability in a dosedependent manner after just 24 h treatment, compared to amiodarone and OA. Liraglutide co-treatment improved cell viability and the effect was greater after a longer period of treatment, especially in tamoxifen-treated cells for almost 30% (p

Češnjak (Allium sativum) ima malo kalorija, bogat je vitaminom C, vitaminom B6 i manganom, te sadrži količine različitih hranjivih sastojaka u tragovima. Brojne studije su pokazale kako smanjuje veličinu čira na želucu, ukupnu kiselost te količinu želučanog soka. Cilj našeg istraživanja bio je smanjiti oksidativno oštećenje nastalo primjenom žučne soli (NaT-a) u ljudskoj želučanoj staničnoj liniji (AGS) uz primjenu inhibitora protonske pumpe lansoprazola (LPZ) te antikoksidativnim djelovanjem češnjakova ulja. Stanice su prethodno tretirane LPZ-om i rastućim koncentracijama češnjaka, te potom izložene 4mM NaT-u. Sposobnost češnjakova ulja u zaštiti AGS stanica od oštećenja izazvanog NaT-om procijenjena je određivanjem koncentracije glutationa (GSH) i ukupnom količinom prostaglandina (PGE2) koristeći ELISA kit, preživljenje AGS stanica brojanjem stanica Neubauerova hemocitometrom te genska ekspresija superoksid dismutaze (SOD) i Nuclear Factor Kappa B Subunit 2 (NFKB2) RT PCR-om. Stanice prethodno tretirane LPZ-om i češnjakovim uljem pokazale su značajno preživljenje u usporedbi s odgovarajućom kontrolom. Predtretman uljem češnjaka povećava proizvodnju PGE2 i suzbija iscrpljivanje GSH. Tretman stanica Nat-om smanjuje ekspresiju SOD-a te NFKB2-a, dok je kod predtretmana uljem češnjaka i LPZ-om dokazana pozitivna korelacija. Predtretman češnjkovim uljem pokazao je gastroprotektivni učinak, premda su potrebni daljnji eksperimenti kako bi se u potpunosti rasvijetlila njegova zaštitna uloga u ulkusnoj bolesti.

Proton pump inhibitors, such as lansoprazole (LPZ) are gold standard in the treatment of ulcer disease. However, various studies have shown effectiveness of garlic extracts (GE) in the treatment of ulcer disease. The aim was to determine inhibition of sodium taurocholate (NaT) caused oxidative stress in human gastric cell line (AGS) by treatment of GE and LPZ. Cells were pre- treated with LPZ and increasing concentrations of GE. The ability of GE to protect the AGS cells against the damage induced by NaT was evaluated by glutathione (GSH) and prostaglandin E(2) content by ELISA, AGS proliferation by cell counting and by expression of superoxide dismutase (SOD) and factor kappa B subunit 2 (NFKB2) by RT PCR. Cells pre-treated with LPZ (p

Background: Cancer poses a major public health problem worlwide. Breast cancer (BC) is the second most common cancer diagnosed in women in Croatia. Anastrozole has demonstrated a longer disease-free survival and better overall safety in the adjuvant treatment of BC. Single nucleotide polymorphism (SNP) in genes encoding drug metabolizing enzymes could have key role in the individual response to anastrozole therapy and drug safety. Objective: To investigate rates of CYP3A4*1B, CYP3A5*3 and UGT1A4*2 SNPs significant in anastrozole metabolism in BC women in Croatia and its potential correlation to anastrozole induced side effects. Methods: 82 postmenopausal women with estrogen receptor (ER)-positive BC treated with anastrozole and 44 BC controls before therapy were included in the study. DNA for SNPs was genotyped by TaqMan RT-PCR and bone mineral density (BMD) was measured by dual-energy x-ray absorptiometry (DXA). Results: Mutant G allele homozygotes were predominant 88%, and homozygotes for the wild type A allele of CYP3A5*3 were not found. Furthermore wild type homozygotes of CYP3A4*1B were predominant with 94% , and mutant homozygotes were not detected. CYP3A4*1B and CYP3A5*3 SNPs were in 84.3% linkage disequilibrium and 95.1% in group treated with anastrozole and without treatment, respectively. Total hip and femoral neck BMD were lower in the group treated with anastrozole. However, potential correlation of BMD changes induced by anastrozole therapy with prevalence of explored SNPs was not shown. Conclusion: No significant differences in BMD among the groups were confirmed. Although, the mutant CYP3A5*3 SNP was predominant and may result in low anastrozole metabolism.

Garlic (Allium sativum) is one of the oldest cultivated plants with significant dietary and medicinal role. It has been used as a spice, food, and herbal medicine for over 4000 years. Numerous studies have shown the gastroprotective effect of garlic extracts (GE). Therefore, the aim was to establish a cell culture model of ulcer disease using bile salts such as sodium taurocholate (NaT) and to determine inhibition of NaT caused oxidative stress by antioxidant treatment of GE. Human gastric cell line (AGS) was used as a cellular model of ulcer disease. Cells were co-treated with different concentrations of garlic extracts (GE). The cytotoxicity of NaT against AGS cells was determined by cell counting. The oxidative stress was evaluated by expression of superoxide dismutase (SOD) by RT PCR. NaT caused a significant reduction in the viability of AGS. It was demonstrated that cells treated with NaT and co-treated with GE showed significant increase in viability, compared to the cells treated solely with NaT. It has been demonstrated a positive correlation of SOD with antioxidant treatment. Garlic extracts co- treatment showed gastroprotective effect on AGS cell culture model of ulcer disease. Nevertheless, further experiments are needed to confirm protective role of garlic extracts in gastric ulcer disease.