Fluoroquinolones' Biological Activities against Laboratory Microbes and Cancer Cell Lines.

Development of novel derivatives to rein in and fight bacteria have never been more demanding, as microbial resistance strains are alarmingly increasing. A multitude of new fluoroquinolones derivatives with an improved spectrum of activity and/or enhanced pharmacokinetics parameters have been widely explored. Reporting novel antimicrobial agents entails comparing their potential activity to their parent drugs; hence, parent fluoroquinolones have been used in research as positive controls. Given that these fluoroquinolones possess variable activities according to their generation, it is necessary to include parent compounds and market available antibiotics of the same class when investigating antimicrobial activity. Herein, we provide a detailed guide on the in vitro biological activity of fluoroquinolones based on experimental results published in the last years. This work permits researchers to compare and analyze potential fluoroquinolones as positive control agents and to evaluate changes occurring in their activities. More importantly, the selection of fluoroquinolones as positive controls by medicinal chemists when investigating novel FQs analogs must be correlated to the laboratory pathogen inquest for reliable results.

In Vitro and In Vivo Antidiabetic Activity, Phenolic Content and Microscopical Characterization of Terfezia claveryi.

Terfezia claveryi (T. claveryi) is used by traditional healers in the Middle East region to treat several diseases, including diabetes. The present study evaluated the total phenolic and investigated the blood-glucose-lowering potential of different aqueous extracts of this selected truffle using in vitro and in vivo models. The phytochemical profile was examined using UPLC-MS. The macerate and the microwave-assisted extract were the richest in phenolic compounds. All T. claveryi extracts exhibited a remarkable α-glucosidase inhibitory effect in vitro, with an IC50 of 2.43, 3.26, 5.18 and 3.31 mg/mL for the aqueous microwave-assisted extract macerate, infusion and decoction, respectively. On the other hand, in the high-fat diet alloxan-induced diabetic mice model, all tested crude aqueous extracts exhibited a significant antihyperglycemic activity (p < 0.05). Four hours after the administration of the 250 mg/kg dose, the macerate was able to induce a 29.4% blood-glucose-lowering effect compared to a 24.8% reduction induced by the infusion, which was sustained for a further two hours. The hypoglycemic effect (29.3% and 32.4%) was also recorded six hours after the administration of the single dose 500 mg/kg of the macerate and the infusion, respectively. Truffle extracts exhibited antidiabetic activity both in vitro and in vivo, providing a rationale for the traditional use as a natural hypoglycemic.

Simple and rapid peptide nanoprobe biosensor for the detection of Legionellaceae.

This study demonstrates the development of a sensitive, specific, and quantitative peptide-based nanoprobe prototype assay for the detection of Legionellaceae in a simple way and in a short time. In this work, proteases present in the culture supernatants of Legionella spp. were used as a biomarker. Fluorogenic peptide substrates, specific to Legionella strains culture supernatant proteases, were identified. Peptidases produced a significant increase in the fluorescence intensity following the cleavage of the dipeptide fluorogenic substrates. The specific substrates were identified and coupled with carboxyl-terminated nano-magnetic particles (NMPs). On the other hand, the C-terminal was conjugated with the cysteine residue to covalently integrate with a gold sensing platform via the Au-S linkage. Four different sensors were fabricated from the four specific substrates, which were treated with the protesase of six different species of Legionella. In the presence of specific protease, the peptide sequence is digested and the magnetic nanobeads moved out of the gold surface, resulting in the apparence of gold color. One of the nanoprobes sensitivity detects as low as 60 CFU mL-1 of Legionella anisa, Legionella micdadei, and Fluoribacter dumoffii. The cross-reactivity of the sensors was tested using other closely associated bacterial species and no significant cross-reactivity of the sensors was found. It is envisaged that this assay could be useful for screening purposes or might be supportive for the fast and easy detection of Legionella protease activity for water monitoring purposes.

Plants Secondary Metabolites as Blood Glucose-Lowering Molecules.

Recently, significant advances in modern medicine and therapeutic agents have been achieved. However, the search for effective antidiabetic drugs is continuous and challenging. Over the past decades, there has been an increasing body of literature related to the effects of secondary metabolites from botanical sources on diabetes. Plants-derived metabolites including alkaloids, phenols, anthocyanins, flavonoids, stilbenoids, saponins, tannins, polysaccharides, coumarins, and terpenes can target cellular and molecular mechanisms involved in carbohydrate metabolism. In addition, they can grant protection to pancreatic beta cells from damage, repairing abnormal insulin signaling, minimizing oxidative stress and inflammation, activating AMP-activated protein kinase (AMPK), and inhibiting carbohydrate digestion and absorption. Studies have highlighted many bioactive naturally occurring plants' secondary metabolites as candidates against diabetes. This review summarizes the current knowledge compiled from the latest studies published during the past decade on the mechanism-based action of plants-derived secondary metabolites that can target various metabolic pathways in humans against diabetes. It is worth mentioning that the compiled data in this review will provide a guide for researchers in the field, to develop candidates into environment-friendly effective, yet safe antidiabetics.

Fluoroquinolones structural and medicinal developments (2013-2018): Where are we now?

Fluoroquinolones are considered one of the widely utilized antibiotics, specifically, with the recent development of newer generations such as moxifloxacin, delafloxacin and finafloxacin which possess a wider spectrum of activity and improved bioavailability. However, alarming increase in bacterial resistance reports worldwide incites medicinal chemists to employ guided-biologically-oriented-synthesis-strategies toward the modulation of quinolones derivatives activity. At present, quinolone scaffold diverse biological activities prompted the fortuitous discovery of novel antimicrobial and anticancer derivatives. Herein, the opponent arguments (2013-2015) regarding the structure activity relationships of fluoroquinolones' antibacterial, anticancer, antifungal activities are inscribed.

Portable paper-based colorimetric nanoprobe for the detection of Stachybotrys chartarum using peptide labeled magnetic nanoparticles.

A colorimetric assay is presented for the detection of Stachybotrys chartarum proteases as biomarkers. The assay comprises a gold film acting as solid support and carrying an immobilized peptide substrate that is specific for S. chartarum protease. The substrate was conjugated to black magnetic nanoparticles (MNPs) to form a monolayer on the gold film. Hence, detection nanoprobe is black. If, however, the peptide-MNP fragments are cleaved by S. chartarum proteases present in a sample, the golden color of the detecting nanoprobe becomes apparent so that positive visual readout is enabled. The method was applied to the determination of S. chartarum in (spiked) environmental samples. The limit of detection ranges from 10 to 100 spores·mL-1 depending on the kind of sample (culture, dust, mold and soil). Assay specificity was examined for Aspergillus flavus, Fusarium solani. Penicillin chrysogenum, and Saccharomyces cerevisiae, and negative readouts were observed visually for all samples, except for those also containing S. chartarum. Graphical abstract Schematic presentation of S. chartarum colorimetric nanoprobe.

Design, synthesis, and biological evaluation of 1,8-naphthyridine glucosamine conjugates as antimicrobial agents.

In the quest for discovering potent antimicrobial agents with lower toxicity, we envisioned the design and synthesis of nalidixic acid-D-(+)-glucosamine conjugates. The novel compounds were synthesized and evaluated for their in vitro antimicrobial activity against Gram positive bacteria, Gram negative bacteria and fungi. Cytotoxicity using MTT assay over L6 skeletal myoblast cell line, ATCC CRL-1458 was carried out. In vitro antimicrobial assay revealed that 1-ethyl-7-methyl-4-oxo-N-(1,3,4,6-tetra-O-acetyl-2-deoxy-D-glucopyranose-2-yl)-[1,8]-naphthyridine-3-carboxamide (5) and 1-ethyl-7-methyl-4-oxo-N-(2-deoxy-D-glucopyranose-2-yl)-[1,8]-naphthyridine-3-carboxamide(6) possess growth inhibitory activity against resistant Escherichia coli NCTC, 11954 (MIC 0.1589 mM) and Methicillin resistant Staphylococcus aureus ATCC, 33591 (MIC 0.1589 mM). Compound (5) was more active against Listeria monocytogenes ATCC 19115 (MIC 0.1113 mM) in comparison with the reference nalidixic acid (MIC 1.0765 mM). Interestingly, compound (6) had potential antifungal activity against Candida albicans ATCC 10231 (MIC <0.0099 mM). Remarkably, the tested compounds had low cytotoxic effect. This study indicated that glucosamine moiety inclusion into the chemical structure of the marketed nalidixic acid enhances antimicrobial activity and safety.

On site visual detection of Porphyromonas gingivalis related periodontitis by using a magnetic-nanobead based assay for gingipains protease biomarkers.

Porphyromonas gingivalis (P. gingivalis) is a pathogen causing periodontitis. A rapid assay is described for the diagnosis of periodontal infections related to P. gingivalis. The method is making use of gingipains, a group of P. gingivalis specific proteases as a detection biomarker. Magnetic-nanobeads were labeled with gingipain-specific peptide substrates and immobilized on a gold biosensing platform via gold-thiol linkage. As a result of this, the color of the gold layer turns black. Upon cleavage of the immobilized substrates by gingipains, the magnetic-nanobeads-peptide fragments were attracted by a magnet so that the golden surface color becomes visible again. This assay is highly sensitive and specific. It is capable of detecting as little as 49 CFU·mL-1 of P. gingivalis within 30 s. Examination of periodontitis patients and healthy control saliva samples showed the potential of the assay. The simplicity and rapidity of the assay makes it an effective point-of-care device. Graphical abstract Schematic of the assay for the detection of P. gingivalis proteases as one of the promising biomarkers associated with periodontal diseases.

Physicochemical characterization of emulgel formulated with SepineoP 600, SepineoSE 68 and cosolvent mixtures.

The combined properties of SepineoP 600 (S600), a self-gelling dispersion and SepineoSE 68 (M68), a natural liquid crystal forming surfactant, were utilized in the development of emulgel base for topical application. The emulgels were prepared in water alone or combined with propylene glycol (PG), polyethylene glycol 400 (PEG400) and glycerol (G) as cosolvents. Emulgels were characterized for their optical and flow behavior. Two model drugs: caffeine (CF) and methylparaben (MP) were used in the evaluation of drug permeation across the stratum corneum (SC). The results showed that emulgel prepared using 70% PG:water (1:1) and 30% S600 has the best flow behavior compared to other cosolvents. Also the permeability coefficient of CF was found to be higher than that of MP and the addition of 3% M68 improved the physical stability of the emulgel, but it did not affect the drug diffusion profile.

Active educational intervention as a tool to improve safe and appropriate use of antibiotics.

Misconception about antibiotics use among the public has been widely outlined to be a main reason for inappropriate use of antibiotics including failure to complete treatment, skipping of doses, re-use of leftover medicines and overuse of antibiotics. The study was devised to evaluate whether education might be a potential strategy to promote safer use of antibiotics and reducing self-medication. Two hundred seventy one adults were asked to complete two questionnaires; a pre and posteducation. The questionnaires comprised of three parts consisting of 17 statements assessing the knowledge on: appropriate use, safe use and resistance of antibiotics. Knowledge score was estimated by calculating the percentage of correct responses. The mean (SD) knowledge score pre-education was 59.4% (20.3). However, posteducation the score was 65.9% (17.9), p < 0.001(t-test). Knowledge scores were classified as poor, adequate and good. Posteducation, participants within poor and adequate knowledge categories were significantly shifted to the good category describing better knowledge, McNemar-χ2 = 28.7, df = 3, p < 0.001. It is concluded that using tailored education material targeting antibiotic need and use with a major aim of improving the public knowledge about antibiotics can be an effective and feasible strategy. This pilot study could be considered as the starting point for a wider scale public educational intervention study and national antibiotic campaign. However, the improvement in participant's knowledge might not reflect an actual change in antibiotics-seeking behaviour or future retention of knowledge. Future research should seek to assess the impact of education on participant's behaviour.

Colorimetric Assay for the Detection of Typical Biomarkers for Periodontitis Using a Magnetic Nanoparticle Biosensor.

Periodontitis is a chronic disease which affects at least 10% of the population. If untreated, periodontitis can lead to teeth loss. Unfortunately, current diagnostic tests are limited in their sensitivity and specificity. In this study, a novel multiplex hand-held colorimetric diagnostic biosensor, using two typical inflammatory salivary biomarkers, Human Neutrophil Elastase (HNE) and Cathepsin-G, was constructed as proof of concept to potentially detect periodontitis. The biosensing method was based on the measurement of proteolytic activity using specific proteases probes. These probes consist of specific proteases substrates covalently bound to a magnetic bead from one end and to the gold sensor surface by the other end. When intact, this renders the golden sensor black. Upon proteolysis, the cleaved magnetic beads will be attracted by an external magnet revealing the golden color of the sensor surface observable by the naked eye. The biosensor was capable of specific and quantitative detection of HNE and Cathepsin-G in solution and in spiked saliva samples with a lower detection limit of 1 pg/mL and 100 fg/mL for HNE and Cathepsin-G, respectively. Examination of periodontitis patients' sample and a healthy control showed the potential of the multiplex biosensor to detect the presence of HNE and Cathepsin-G activity in situ. This approach is anticipated to be a useful biochip array amenable to low-cost point-of-care devices.

Design, Synthesis and in Vivo Evaluation of Novel Glycosylated Sulfonylureas as Antihyperglycemic Agents.

Sulphonylurea compounds have versatile activities such as antidiabetic, diuretic, herbicide, oncolytic, antimalarial, antifungal and anticancer. The present study describes the design, synthesis and in vivo testing of novel glycosylated aryl sulfonylurea compounds as antihyperglycaemic agents in streptozocine-induced diabetic mice. The rational for the introduction of the glucosamine moiety is to enhance selective drug uptake by pancreatic ß-cells in order to decrease the cardiotoxic side effect commonly associated with sulfonylurea agents. 2-Deoxy-2-(4-chlorophenylsulfonylurea)-D-glucopyranose was found to be the most potent antihyperglycaemic agents among the synthesized compounds in diabetic mice. This investigation indicates the importance of this novel class as potential antihyperglycaemic agents.

Glucosamine substituted sulfonylureas: IRS-PI3K-PKC-AKT-GLUT4 insulin signalling pathway intriguing agent.

Normally, skeletal muscle accounts for 70-80% of insulin-stimulated glucose uptake in the postprandial hyperglycemia state. Consequently, abnormalities in glucose uptake by skeletal muscle or insulin resistance (IR) are deemed as initial metabolic defects in the pathogenesis of type 2 diabetes mellitus (T2DM). Globally, T2DM is growing in exponential proportion. The majority of T2DM patients are treated with sulfonylureas in combination with other drugs to improve insulin sensitivity. Glycosylated sulfonylureas (sulfonylurea-glucosamine analogues) are modified analogues of sulfonylurea that have been previously reported to possess antidiabetic activity. The aim of this study was to evaluate the impact of glycosylated sulfonylureas on the insulin signalling pathway at the molecular level using L6 skeletal muscle cell (in vitro) and extracted soleus muscle (ex vivo) models. To create an in vitro model, insulin resistance was established utilizing a high insulin-glucose approach in differentiated L6 muscle cells from Rattus norvegicus. Additionally, for the ex vivo model, extracted soleus muscles, adult Sprague-Dawley rats were subjected to a solution containing 25 mmol L-1 glucose and 100 mmol L-1 insulin for 24 hours to induce insulin resistance. After insulin resistance, compounds under investigation and standard medicines (metformin and glimepiride) were tested. The differential expression of PI3K, IRS-1, PKC, AKT2, and GLUT4 genes involved in the insulin signaling pathway was evaluated using qPCR. The evaluated glycosylated sulfonylurea analogues exhibited a significant increase in the gene expression of insulin-dependent pathways both in vitro and ex vivo, confirming the rejuvenation of the impaired insulin signaling pathway genes. Altogether, glycosylated sulfonylurea analogues described in this study represent potential therapeutic anti-diabetic drugs.

A cross-sectional study on adult lifestyle habits during the COVID-19 pandemic.

COVID-19 has spread and developed into a pandemic disease, forcing countries to impose challenging protocols and lockdowns. This study assessed shopping, food consumption behavior, and feelings in Jordan and several Arab countries during the COVID-19 pandemic. A cross-sectional web-based survey among the Middle East population was conducted using an online questionnaire between July and September 2022. Participants were requested to answer a standardized and validated structured questionnaire. Demographic information, shopping behavior information, and mental health data were requested. A total of 542 respondents were included in the study. During COVID-19 quarantine, participants (68.6%) reported decreased shopping frequency and buying more food than usual (37.5%). Cereals and legumes were the primary food types stored by participants (76.9%). Multiple logistic regression revealed the age of the participant as a significant factor affecting storing of food (being ≤ 25 years old (OR = 0.456, p value = 0.038)). 75.7% of female participants eat less frequently in restaurants than usual. In contrast, among males, 48.5% reported that they eat at restaurants less frequently than usual. The country of residency and gender were the significant factors affecting negative feelings and emotions. Participants in countries other than Jordan had a higher negative feeling score (Beta = 0.086, p value = 0.042). Furthermore, females had a higher negative feeling score (Beta = -0.128, p value = 0.003) as the negative feelings score for females was 3.58 (SD = 5.443). On the other hand, it was 2.10 (SD = 5.091) for males. The COVID-19 pandemic has altered Jordanians' attitudes, shopping, and food consumption habits. Although positive behaviors have improved, such as shopping less frequently, eating home-cooked meals, and dining with family, frequent snacking and food storage have increased. Finally, public awareness of shopping and food consumption habits should be promoted.

Rapid Colorimetric Quantita2tive Portable Platform for Detection of Brucella melitensis Based on a Fluorescence Resonance Energy Transfer Assay and Nanomagnetic Particles.

Brucellosis is a bacterial zoonotic disease that requires major attention for both health and financial facilities in many parts of the world including the Mediterranean and the Middle East. The existing gold standard diagnosis relies on the culturing technique, which is costly and time-consuming with a duration of up to 45 days. The Brucella protease biosensor represents a new detection approach that will lead to low-cost point-of-care devices for sensitive Brucella detection. In addition, the described diagnostic device is portable and simple to operate by a nurse or non-skilled clinician making it appropriate for the low-resource setting. In this study, we rely on the total extracellular protease proteolytic activity on specific peptide sequences identified using the FRET assay by high-throughput screening from the library of peptide (96 short peptides such as dipeptides and tripeptides) substrates for Brucella melitensis (B. melitensis). The B. melitensis-specific protease substrate was utilized in the development of the paper-based colorimetric assay. Two specific and highly active dipeptide substrates were identified (FITC-Ahx-K-r-K-Ahx-DABCYL and FITC-Ahx-R-r-K-Ahx-DABCYL). The peptide-magnetic bead nanoprobe sensors developed based on these substrates were able to detect the Brucella with LOD as low as 1.5 × 102 and 1.5 × 103 CFU/mL using K-r dipeptide and R-r dipeptide substrates, respectively, as the recognition element. The samples were tested using this sensor in few minutes. Cross-reactivity studies confirmed that the other proteases extracted from closely related pathogens have no significant effect on the sensor. To the best of our knowledge, this assay is the first assay that can be used with low-cost, rapid, direct, and visual detection of B. melitensis.

Ultra-rapid colorimetric assay for protease detection using magnetic nanoparticle-based biosensors.

Sensitive protease detection methods often require time-consuming techniques and expensive instrumentation. To overcome this limitation, a novel, simple, sensitive and selective colorimetric detection approach was developed. This biosensing configuration was validated by the use of prostate specific antigen (PSA) protease as a model target. In this method, proteolytically active PSA capable of cleaving PSA substrates caused the release of black-colored magnetic carrier complexes, exposing the gold color sensor surface visible to the naked eye. The assay showed excellent sensitivity as well as specificity, capable of discriminating between different types of protease targets. The biosensor was able to quantitatively detect different PSA concentrations with a detection limit as low as 10 ng mL(-1). The sensor offers the possibility of developing a wash-less and cost-effective point-of-care device due to the simplicity of the probe immobilization process and the elimination of labeling and reporter molecules during the biosensing step.

Complementary and alternative medicine utilization by a sample of infertile couples in Jordan for infertility treatment: clinics-based survey.

BACKGROUND: Although there is little information available to quantify the use of complementary and alternative medicine (CAM), growing evidence suggests that CAM prevalence among patients seeking infertility treatment is increasing worldwide. There are many products available on the market and many infertile patients demand information about CAM from their health care providers. This paper investigates the prevalence of CAM use among infertile couples in Jordan. Additionally, trends and factors contributing to CAM use for infertility treatment among these couples have been evaluated. METHODS: A face-to-face questionnaire inquiring demographic information, use of CAM for medical conditions, in general, and types of CAM used for infertility treatment, in specific, was completed by one thousand twenty one infertile patients attending at two types of facilities; in vitro Fertilization (IVF) centers at both public and private hospitals and infertility private clinics. Both types of facilities were distributed in different areas of Amman, the capital city of Jordan. The study was conducted between May and August 2012. RESULTS: Our results show that CAM therapies for infertility treatment were encountered in 44.7% of the study sample. The vast majority of CAM users were females. The most commonly used CAM therapies were herbs and spiritual healing. A clear correlation between the use of CAM for infertility versus the use of CAM for other chronic medical conditions has been found. CONCLUSIONS: The prevalence of CAM use for infertility treatment in Jordan is relatively high, particularly among young females, well educated and with a low income, in consistence with the studies reported elsewhere. Herbs and spiritual healing are widely used among patients in adjunct to conventional medical interventions. As CAM use is prevalent among patients, there is a clear need for health providers to become more aware of this phenomenon and for further research in this field.

Development of Rapid Aptamer-Based Screening Assay for the Detection of Covid-19 Variants.

The development of a colorimetric severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) detection assay with the WHO published ASSURED criteria is reported, in which the biosensor should have the following characteristics of (i) being affordable for low-income communities, (ii) sensitive, (iii) specific, (iv) user-friendly to be used by non-skilled personnel, (v) rapid and robust, (vi) equipment-free, and (vii) delivered to the end-users as a simple and easy to use point-of-care tool. Early viral infection detection prevents virus spread and controls the epidemic. We herein report the development of a colorimetric assay in which SARS-COV-2 variants can be detected by colorimetric observation of color on the sensing cotton swab surface. Using the developed biosensor assay, it is possible to discriminate between the various SARS-CoV-2 variants with a LOD of 100 ng/mL within 4 min including sample preconcentration and incubation step. The results illustrated the development of a SARS-CoV-2 colorimetric biosensor that can be mass produced, with low-reagent cost, and can be read-out visually in the field by nonskilled personnel.

Antibiotic-Lysobacter enzymogenes proteases combination as a novel virulence attenuating therapy.

Minimizing antibiotic resistance is a key motivation strategy in designing and developing new and combination therapy. In this study, a combination of the antibiotics (cefixime, levofloxacin and gentamicin) with Lysobacter enzymogenes (L. enzymogenes) bioactive proteases present in the cell- free supernatant (CFS) have been investigated against the Gram-positive methicillin-sensitive Staphylococcus aureus (MSSA), methicillin-resistant Staphylococcus aureus (MRSA) and the Gram-negative Escherichia coli (E. coli O157:H7). Results indicated that L. enzymogenes CFS had maximum proteolytic activity after 11 days of incubation and higher growth inhibitory properties against MSSA and MRSA compared to E. coli (O157:H7). The combination of L. enzymogenes CFS with cefixime, gentamicin and levofloxacin at sub-MIC levels, has potentiated their bacterial inhibition capacity. Interestingly, combining cefixime with L. enzymogenes CFS restored its antibacterial activity against MRSA. The MTT assay revealed that L. enzymogenes CFS has no significant reduction in human normal skin fibroblast (CCD-1064SK) cell viability. In conclusion, L. enzymogenes bioactive proteases are natural potentiators for antimicrobials with different bacterial targets including cefixime, gentamicin and levofloxacin representing the beginning of a modern and efficient era in the battle against multidrug-resistant pathogens.