Docking Study to Predict the Efficacy of Phosphatidylinositol 3-Kinase α Inhibitors

The phosphatidylinositol 3-kinase (PI3K) family comprises lipid kinases that cross-link signals between living cells and their surroundings. PI3Ks are classified into several groups and isoforms with specific characteristics and functions. Genes encoding PI3Ks are mutated in several types of cancer, and their isoforms have varying capacity in promoting cell signaling and cancer progression. Many compounds have been introduced as PI3Kα inhibitors, but not all of them have the same inhibitory effects. For successful PI3K-related biomedical experiments, it is vital to select the most specific and potent compounds with the highest inhibitory effects for targeting this kinase. In this study, we investigate 28 well-recognized PI3Kα inhibitors through predicting their specificity and potency using the docking software AutoDock Vina. Our data showed that PF 05212384 had the highest docking score (−9.2 kcal/mol), and 3-methyladenine had the lowest docking score (−4.8 kcal/mol). Our data also showed different types of interactions and bonds formed between the inhibitors and protein residues. In conclusion, PF 05212384 and AZD 6482 compounds are the best candidates for targeting PI3Kα. In addition to hydrophobic interactions in the PI3Kα binding pocket, the formation of hydrogen bonds between these inhibitors and binding pocket residues was confirmed

Role of Laser Produced Silver Nanoparticles in Reversing Antibiotic Resistance in Some MultidrugResistant Pathogenic Bacteria

Silver nanoparticles (Ag NPs) were produced through nanosecond laser in deionized water. These nanoparticles were characterized by UV–VIS spectrometer and transmission electron microscopy. VITEK®2 compact system was used to identify Escherichia coli (ESBL strain) and Staphylococcus aureus (MRSA strain) as multidrug-resistance (MDR) bacteria. The antibacterial activity of Ag NPs, ampicillin (AMP), and their combinations was tested against both bacterial isolates through standard microbiological culturing techniques. Our data show that both of E. coli and S. aureus were highly resistant to AMP. Ag NPs alone reduced growth in both bacterial isolates considerably. Growth declined drastically in both bacteria when AMP was used in combination with Ag NPs. The minimal inhibitory concentration of combined agents for E. coli was 20 µg/ml Ag NPs + 1 mg AMP/ml and for S. aureus was 10 µg/ml Ag NPs + 1 mg AMP/ml. The results show that the Ag NPs have great potentials in enhancing the antimicrobial activities of drugs that used to be ineffective against MDR bacteria. Administering combinations of antibiotic(s) with AgNPs may help in treating patients suffering from infections caused by MDR bacteria. Further in vivo and in vitro investigations are required to evaluate the side effects of these combinations

Bacterial Profile and Antimicrobial Susceptibility of Isolates Recovered from Lower Respiratory Tract Infection for Patients in Rizgary Hospital, Erbil

Recognition of etiologies of lower respiratory tract infection (LRTI) may help in delivering effective treatment options and circumvent emergence of antibiotic resistance. This study was carried out to uncover bacterial profile and antibiotic sensitivity patterns among 310 LRTI patients attended Rizagary Hospital between January 2014 to December 2016. Standard laboratory techniques were applied in collecting, processing, and culturing sputum and bronchial wash specimens. VITEK® 2 compact systems were used to identify bacteria and their antibiotic sensitivity patterns. Results showed that Streptococcus parasanguinis and Acinetobacter baumannii were the most abundant gram-positive and gram-negative bacteria (GPB & GNB), respectively, isolated from sputum specimens. From bronchial wash specimens, only GNB were detected and Serratia marcescens was the most abundant one. Antibiotic sensitivity tests revealed that Streptococcus parasanguinis was the most resistant GPB and Acinetobacter baumannii was the most resistant GNB. Sputum recovered GPB were highly resistant to Ampicillin, Erythromycin, Levofloxacin, Trimethoprim/Sulfamethoxazole, and Tetracycline. Bronchial wash recovered GNB were highly resistant to Ampicillin, Minocycline, Pefloxacin, Piperacillin, and Ticarcillin. In conclusion, LRTIs are mainly associated with GNB rather than GPB. The recovered Streptococcus parasanguinis and Acinetobacter baumannii were found to be multidrug-resistant pathogens. Ampicillin was ineffective against any of recovered pathogenic bacteria

Categorization of Bacterial Pathogens Present in Infected Wounds and their Antibiotic Resistance Profile Recovered from Patients Attending Rizgary Hospital-Erbil

Wound infection with antibiotic-resistant bacteria can extend a patients’ debility and increase the expense of treatment in the long term; therefore, careful management of patients with wound infections is necessary to avoid complications. The usage of antimicrobial agent is a major factor in resistance development. This study aims to understand the causes of wound infections, as well as the criteria for diagnosing them for more sensible antibiotic prescribing. Samples from 269 wound patients were collected, and cultured for bacterial growth. Gram stain technique, bacterial identification via VITEK 2 compact system were investigated in this study. Gram negative bacteria accounted for 59.15% of the total isolates, while pathogenic gram positive bacteria accounted for 40.85% of total isolates. Escherichia coli and Pseudomonas aeruginosa are the dominant pathogenic gram negative bacteria in wounds, while Staphylococcus aureus, and Staphylococcus epidermidis are the dominant pathogenic gram positive bacteria. Pseudomonas aeruginosa showed 100% resistance to the majority of antibiotic tested, including Ampicillin, Amoxicillin/Clavulanic Acid, Aztreona, Ceftriaxone, and others. Staphylococcus aureus and Staphylococcus epidermidis are 100% resistant to Ampicillin, Ceftriaxone, and Cefotaxime. For more efficient antibiotic prescriptions, the causative microorganisms, and their current susceptibility patterns need to be mandated for testing before prescribing any antibiotics to patients. Prescriptions are frequently based solely on general information about the antibiotic's function, rather than on individual response variation to the pathogen and the antibiotic. Particularly when the common pathogens in this study show multidrug resistance in wounds

mTOR modulates resistance to gemcitabine in lung cancer in an MTORC2 dependent mechanism.

Lung cancer has a poor prognosis partly due to a lack of response to treatments such as the chemotherapy drug gemcitabine. Combinations of chemotherapy drugs with signal transduction inhibitors may be more effective treatments. In this study we have investigated the impact of targeting the mTOR signalling pathway on the efficacy of gemcitabine in different cancer cell lines. Time-lapse microscopy, immuno-staining, and western blot techniques were used to evaluate the efficacy of applied treatments either in measuring phosphorylation levels of mTOR down-stream targets or in tracking down the fate of targeted cells. Reactive oxygen species and relative levels of protein phosphorylation were also quantified. For comparison between treated groups t-test and analysis of variance test were applied. Our data showed that mTORC1 has no role in sensitising A549 lung cancer cells to gemcitabine. However, targeting mTORC1/2 with the pharmacological inhibitor torin1 or by over-expressing Deptor, the negative regulator of mTOR signalling, sensitised these cells to gemcitabine. Silencing mTORC2, but not mTORC1, induced apoptosis and significantly improved the apoptosis-inducing effects of gemcitabine. Results also suggest that Rictor is required to maintain cell survival through modulating p38α, ERK1/2, RSK1/2/3 and the transcription factor STAT3. Multiple cell line comparisons revealed that PANC-1 pancreatic cancer cells were also sensitive to mTOR inhibition, but MCF7 breast cancer, MCF10A breast epithelial and H727 lung cancer cell lines were more resistant to the treatment. Inhibition of mTORC2 may have benefits in the treatment of gemcitabine resistant cancers, and the genetic background of the cell line may determine its response to mTOR inhibition. [Abstract copyright: Copyright © 2021 Elsevier Inc. All rights reserved.

Prevalence and antimicrobial susceptibility of bacterial pathogens isolated from urine specimens received in rizgary hospital — Erbil

Background: Urinary tract infection (UTI) is a common health-associated problem worldwide. Like other medical conditions, UTI patients may suffer from poor treatment outcomes due to the emergence of antimicrobial resistance. Determining patterns of antimicrobial susceptibility in uropathogens will guide physicians to choose the best antibiotics for treating affected patients. In this project we aimed to evaluate the frequencies of pathogens associated with UTI and their antimicrobial susceptibility patterns. Methods: This study was conducted on 2692 urine samples of patients visited Rizgary Teaching Hospital in Erbil city. Aerobic bacterial growth identification and antimicrobial susceptibility tests were performed using VITEK®2 compact system. Results: Our data show that more than 20% of all studied samples were negative for bacterial growth; only 16.72% of them were pathogenic bacteria in which 82.44% of them were Gram negative bacteria (GNB) and the rest were Gram positive bacteria (GPB). Escherichia coli was the most frequent, and Acinetobacter baumannii was the most resistant GNB. Staphylococcus haemolyticus was the most frequent, and Enterococcus faecalis was the most resistant GPB. In general GNB were highly resistant to Ticarcillin and Cefepime, and GPB were also resistant to Ticarcillin, and Tigecycline antibiotics. Conclusions: The amount of negative culture growth indicates that symptoms only based diagnosis for UTI detection is unreliable. E. coli is the most UTI related pathogen, E. faecalis and A. baumannii were among highly antibiotic resistant bacteria. Finally, since many of GNG and GPB isolates were resistant to several antibiotics, there might be a high possibility for multi drug resistant among local population in Erbil. Keywords: Antimicrobial susceptibility, Bacterial profile, E. coli, Multi drug resistant, Urinary Tract Infection (UTI

Fenretinide induces apoptosis and synergises the apoptosis inducing effect of gemcitabine through inhibition of key signalling molecules involved in A549 cell survival in in silico and in vitro analyses.

Fenretinide is a synthetic retinoid compound, which induces apoptosis via generating reactive oxygen species (ROS) and modulating PI3K/Akt/mTOR signalling pathway. We hypothesise that fenretinide's mechanism of action in triggering apoptosis may involve other targets, beside mTOR signalling pathway and it may augment apoptosis inducing effects of chemotherapeutic drugs in lung cancer. Time-lapse microscopy and Western blotting were used to evaluate apoptosis and apoptotic marker cleaved-Caspase 3 in A549 cells. Relative levels of protein phosphorylation and ROS were quantified by Human Phospho-Kinase Array Kit and CellROX® Green Reagent, respectively. Docking and simulation analyses of proteins and fenretinide interactions were identified and visualised by Discovery Studio Visualizer and AutoDock Vina software. Our results showed that fenretinide induced apoptosis in a dose dependant manner and combinations of fenretinide (5 μg/mL) and gemcitabine (1, 2, 4, 8 and 16 μg/mL) synergistically enhanced apoptosis in A549 cells. Fenretinide caused significant increase of cleaved-Caspase 3, de-phosphorylated p-S473 of Akt and failed to inhibit mTORC1 downstream targets. In silico results revealed that Akt required the lowest energy (-10.2 kcal/mol) to interact with fenretinide in comparison with other proteins. In conclusion, Akt may be exploited as a good target for induction of apoptosis in A549 cells and fenretinide has great potentials to fulfil this task. The mechanism by which fenretinide boosts the apoptosis inducing effects of gemcitabine, which is likely expected to be via inhibiting mTORC2 downstream targets. However, docking investigation revealed that fenretinide lacks specificity as it may also interact with several secondary targets beside Akt