Repurposing carvacrol, cinnamaldehyde, and eugenol as potential anti-quorum sensing agents against uropathogenic Escherichia coli isolates in Alexandria, Egypt.

BACKGROUND: Urinary tract infections represent one of the most frequent hospital and community-acquired infections with uropathogenic Escherichia coli (UPEC) being the main causative agent. The global increase in the emergence of multidrug-resistant (MDR) UPEC necessitates exploring novel approaches. Repurposing natural products as anti-quorum sensing (QS) agents to impede bacterial virulence is gaining momentum nowadays. Hence, this study investigates the anti-QS potentials of carvacrol, cinnamaldehyde, and eugenol against E. coli isolated from urine cultures of Egyptian patients. RESULTS: Antibiotic susceptibility testing was performed for 67 E. coli isolates and 94% of the isolates showed MDR phenotype. The usp gene was detected using PCR and accordingly, 45% of the isolates were categorized as UPEC. Phytochemicals, at their sub-inhibitory concentrations, inhibited the swimming and twitching motilities of UPEC isolates, with eugenol showing the highest inhibitory effect. The agents hindered the biofilm-forming ability of the tested isolates, at two temperature sets, 37 and 30 °C, where eugenol succeeded in significantly inhibiting the biofilm formation by > 50% at both investigated temperatures, as compared with untreated controls. The phytochemicals were shown to downregulate the expression of the QS gene (luxS) and critical genes related to motility, asserting their anti-QS potential. Further, the combinatory activity of the phytoproducts with five antibiotics was assessed by checkerboard assay. The addition of the phytoproducts significantly reduced the minimum inhibitory concentrations of the antibiotics and generated several synergistic or partially synergistic combinations, some of which have not been previously explored. CONCLUSIONS: Overall, carvacrol, cinnamaldehyde, and eugenol could be repurposed as potential anti-QS agents, which preferentially reduce the QS-based communication and attenuate the cascades of gene expression, thus decreasing the production of virulence factors in UPEC, and eventually, subsiding their pathogenicity. Furthermore, the synergistic combinations of these agents with antibiotics might provide a new perspective to circumvent the side effects brought about by high antibiotic doses, thereby paving the way for overcoming antibiotic resistance.

Promising biotherapeutic prospects of different probiotics and their derived postbiotic metabolites: in-vitro and histopathological investigation.

BACKGROUND: Probiotics and their derived postbiotics, as cell-free supernatants (CFS), are gaining a solid reputation owing to their prodigious health-promoting effects. Probiotics play a valuable role in the alleviation of various diseases among which are infectious diseases and inflammatory disorders. In this study, three probiotic strains, Lactiplantibacillus plantarum, Lacticaseibacillus rhamnosus, and Pediococcus acidilactici, were isolated from marketed dietary supplements. The antimicrobial activity of the isolated probiotic strains as well as their CFS was investigated. The neutralized CFS of the isolated probiotics were tested for their antibiofilm potential. The anti-inflammatory activity of the isolated Lactobacillus spp., together with their CFS, was studied in the carrageenan-induced rat paw edema model in male Wistar rats. To the best of our knowledge, such a model was not previously experimented to evaluate the anti-inflammatory activity of the CFS of probiotics. The histopathological investigation was implemented to assess the anti-inflammatory prospect of the isolated L. plantarum and L. rhamnosus strains as well as their CFS. RESULTS: The whole viable probiotics and their CFS showed variable growth inhibition of the tested indicator strains using the agar overlay method and the microtiter plate assay, respectively. When tested for virulence factors, the probiotic strains were non-hemolytic lacking both deoxyribonuclease and gelatinase enzymes. However, five antibiotic resistance genes, blaZ, ermB, aac(6')- aph(2"), aph(3'')-III, and vanX, were detected in all isolates. The neutralized CFS of the isolated probiotics exhibited an antibiofilm effect as assessed by the crystal violet assay. This effect was manifested by hindering the biofilm formation of the tested Staphylococcus aureus and Pseudomonas aeruginosa clinical isolates in addition to P. aeruginosa PAO1 strain. Generally, the cell cultures of the two tested probiotics moderately suppressed the acute inflammation induced by carrageenan compared to indomethacin. Additionally, the studied CFS relatively reduced the inflammatory changes compared to the inflammation control group but less than that observed in the case of the probiotic cultures treated groups. CONCLUSIONS: The tested probiotics, along with their CFS, showed promising antimicrobial and anti-inflammatory activities. Thus, their safety and their potential use as biotherapeutics for bacterial infections and inflammatory conditions are worthy of further investigation.

Genomic characterization of colistin-resistant Klebsiella pneumoniae isolated from intensive care unit patients in Egypt.

BACKGROUND: Egypt has witnessed elevated incidence rates of multidrug-resistant Klebsiella pneumoniae infections in intensive care units (ICUs). The treatment of these infections is becoming more challenging whilst colistin-carbapenem-resistant K. pneumoniae is upsurging. Due to the insufficiently available data on the genomic features of colistin-resistant K. pneumoniae in Egypt, it was important to fill in the gap and explore the genomic characteristics, as well as the antimicrobial resistance, the virulence determinants, and the molecular mechanisms of colistin resistance in such a lethal pathogen. METHODS: Seventeen colistin-resistant clinical K. pneumoniae isolates were collected from ICUs in Alexandria, Egypt in a 6-month period in 2020. Colistin resistance was phenotypically detected by modified rapid polymyxin Nordmann/Poirel and broth microdilution techniques. The isolates susceptibility to 20 antimicrobials was determined using Kirby-Bauer disk diffusion method. Whole genome sequencing and bioinformatic analysis were employed for exploring the virulome, resistome, and the genetic basis of colistin resistance mechanisms. RESULTS: Out of the tested K. pneumoniae isolates, 82.35% were extensively drug-resistant and 17.65% were multidrug-resistant. Promising susceptibility levels towards tigecycline (88.24%) and doxycycline (52.94%) were detected. Population structure analysis revealed seven sequence types (ST) and K-types: ST383-K30, ST147-K64, ST17-K25, ST111-K63, ST11-K15, ST14-K2, and ST525-K45. Virulome analysis revealed yersiniabactin, aerobactin, and salmochelin siderophore systems in ˃ 50% of the population. Hypervirulence biomarkers, iucA (52.94%) and rmpA/A2 (5.88%) were detected. Extended-spectrum ß-lactamase- and carbapenemase-producers accounted for 94.12% of the population, with blaCTX-M-15, blaNDM-5, and blaOXA-48 reaching 64.71%, 82.35%, and 82.35%, respectively. Chromosomal alterations in mgrB (82.35%) were the most prevailing colistin resistance-associated genetic change followed by deleterious mutations in ArnT (23.53%, L54H and G164S), PmrA (11.76%, G53V and D86E), PmrB (11.76%, T89P and T134P), PmrC (11.76%, S257L), PhoQ (5.88%, L322Q and Q435H), and ArnB (5.88%, G47D) along with the acquisition of mcr-1.1 by a single isolate of ST525. CONCLUSIONS: In this study, we present the genotypic colistin resistance mechanisms in K. pneumoniae isolated in Egypt. More effective antibiotic stewardship protocols must be implemented by Egyptian health authorities to restrain this hazard and safeguard the future utility of colistin. This is the first characterization of a complete sequence of mcr-1.1-bearing IncHI2/IncHI2A plasmid recovered from K. pneumoniae clinical isolate belonging to the emerging high-risk clone ST525.

Biological effect of bone marrow mesenchymal stem cell- derived extracellular vesicles on the structure of alveolar bone in rats with glucocorticoid-induced osteoporosis.

BACKGROUND: Glucocorticoids are used for the treatment of autoimmune disorders; however, they can elicit several side effects such as osteoporosis. Several approaches can be made to treat glucocorticoid-induced osteoporosis, including the use of stem cells. However, the therapeutic effect of mesenchymal stem cells depends on its released factors, including extracellular vesicles. Extracellular vesicles have been recognized as important mediators of intercellular communication as they participate in many physiological processes. The present study was designed to investigate the effect of bone marrow mesenchymal stem cells derived extracellular vesicles on the structure of alveolar bone in rats with glucocorticoid-induced osteoporosis. METHODS: Thirty adult albino male rats were divided into 3 groups: control group (CG), glucocorticoid-induced osteoporosis (GOG) and extracellular vesicles treated group (ExTG). Rats in the GOG and ExTG groups were injected with methylprednisolone acetate (40 mg/kg) intramuscularly in the quadriceps muscle 3 times per week for three weeks in the early morning. Afterwards, the rats in GOG group received a single vehicle injection (PBS) while each rat in the ExTG group received a single injection of extracellular vesicles (400 µg/kg suspended in 0.2 ml PBS) in the tail vein. Rats were euthanized 1 month after injection. Mandibles were dissected and the molar segments were prepared for histological preparation, scanning electron microscopy (SEM), and energy dispersive x-ray (EDX). RESULTS: Histology and scanning electron microscopyof bone tissue showed alveolar bone loss and bone resorption in the GOG group. while in the ExTG group, alveolar bone demostrated normal bone architecture. EDX showed that calcium percentage in GOG group was lower than ExTG group,which showed no statistically significant difference from the control group. CONCLUSIONS: Extracellular vesicles may be a promising treatment modality in the treatment of bone diseases and in bone regeneration. However, further research is needed before stating that extracellular vesicles s can be used to treat bone disorders especially when translating to humans.

Short-Term Outcomes of Neuromuscular Electrical Stimulation in Critically Ill Patients.

Muscle weakness acquired in the intensive care unit (ICU) adversely affects outcomes of ICU patients. This article reports the short-term respiratory effects of neuromuscular electrical stimulation (NMES) in critically ill patients. Patients were randomly assigned to an intervention group (NMES + conventional physiotherapy) and a control group (sham NMES + conventional physiotherapy). The application of NMES in the intervention group resulted in a significant decrease in the duration of mechanical ventilation and reduced the number of weaning trial failures. Other positive outcomes included reductions in the length of ICU stays and decreased mortality when compared with the control group.

Pathognomonic features of Pasteurella multocida isolates among various avian species in Sharkia Governorate, Egypt.

The present study aimed to isolate Pasteurella multocida (P. multocida) from pulmonary cases in several avian species and then investigate the histopathological features, antimicrobial resistance determinants, virulence characteristics, and risk factors analysis of the isolates in each species in correlation with epidemiological mapping of pasteurellosis in Sharkia Governorate, Egypt. The obtained data revealed a total occurrence of 9.4% (30/317) of P. multocida among the examined birds (chickens, ducks, quails, and turkeys). The incidence rate was influenced by avian species, climate, breed, age, clinical signs, and sample type. Antimicrobial susceptibility testing revealed that all isolates were sensitive to florfenicol and enrofloxacin, while 86.6 and 73.3% of the isolates displayed resistance to amoxicillin-clavulanic acid and erythromycin, respectively. All of the P. multocida isolates showed a multiple-drug resistant pattern with an average index of 0.43. Molecular characterization revealed that the oma87, sodA, and ptfA virulence genes were detected in the all examined P. multocida isolates. The ermX (erythromycin), blaROB-1 (ß-lactam), and mcr-1(colistin) resistance genes were present in 60, 46.6, and 40% of the isolates, respectively. Ducks and quails were the most virulent and harbored species of antimicrobial-resistant genes. These results were in parallel with postmortem and histopathological examinations which detected more severe interstitial pneumonia lesions in the trachea and lung, congestion, and cellular infiltration especially in ducks. Epidemiological mapping revealed that the Fakous district was the most susceptible to pasteurellosis infection. Thus, farmers are recommended to monitor their flocks for signs of respiratory disease, seek veterinary care promptly if any birds are sick, and avoid the random usage of antibiotics. In conclusion, this study presents a comprehensive picture of the risk factors in correlation to the pathognomonic characteristics of P. multocida infection in poultry sectors to help in developing more effective strategies for prevention and control.

An Alternative Approach Using Nano-garlic Emulsion and its Synergy with Antibiotics for Controlling Biofilm-Producing Multidrug-Resistant Salmonella in Chicken.

Surface-growing antibiotic-resistant pathogenic Salmonella is emerging as a global health challenge due to its high economic loss in the poultry industry. Their pathogenesis, increasing antimicrobial resistance, and biofilm formation make them challenging to treat with traditional therapy. The identification of antimicrobial herbal ingredients may provide valuable solutions to solve this problem. Therefore, our aim is to evaluate the potency of nano garlic as the  alternative of choice against multidrug-resistant (MDR) Salmonella isolates using disc diffusion and microdilution assays. Then, checkerboard titration in trays was applied, and FIC was measured to identify the type of interaction between the two antimicrobials. A disc diffusion assay revealed that neomycin was the drug of choice. The range of nano garlic MIC was 12.5-25 µg/ml, while the neomycin MIC range was 32-64 µg/ml. The FIC index established a synergistic association between the two tested drugs in 85% of isolates. An experimental model was used including nano garlic and neomycin alone and in combination against Salmonella infection. The combination therapy significantly improved body productivity and inhibited biofilm formation by more than 50% down regulating the CsgBAD, motB, and sipA operons, which are responsible for curli fimbriae production and biofilm formation in Salmonella serotypes.

Hantzsch condensation reaction as a spectrofluorometric method for determination of alogliptin, an antidiabetic drug, in pure form, tablet form, and human and rat plasma.

To analyze alogliptin in its pharmaceutical dosage forms and human plasma, a sensitive, inexpensive, simple, and precise spectrofluorimetric method was developed and tested. This method was also used to investigate the drug's pharmacokinetic behaviour in the blood of rats. This was based on the Hantzsch reaction, which produces yellowish luminous products that can be detected spectrofluorometrically at 480 and 415 nm for emission and excitation, respectively, when the primary amine group in the examined drug reacts with acetylacetone and formaldehyde. Several experimental parameters that affect the reaction product's development and stability were explored and improved. The curve of fluorescence and concentration for alogliptin was linear in the concentration range 0.05-3.60 µg ml-1 . The proposed approach was validated according to International Council for Harmonization criteria. The method was successfully utilized to evaluate the examined drug in dose formulations and spiked human plasma with high accuracy.

Efficacy of platelet rich plasma on pancreatic injury induced by renal ischemia reperfusion in adult male rats.

Renal ischemia-reperfusion (I-R) injury is the main cause of acute renal failure. Acute pancreatitis is one of the fatal remote lesions that occurs in patients with renal I-R injury. Platelet-rich plasma (PRP) is a hopeful aiding therapy for tissue injuries. Forty adult rats were utilized in this study, ten for PRP preparation and thirty were divided into three groups; Control: subdivided into three equal subgroups, I-R group: exposed to bilateral renal pedicles clamping and I-R+ PRP group: were experienced identical procedures as I-R group then subcutaneously (S.C) injected with 0.5 ml PRP two times weekly for three weeks. Blood samples were taken for detection of serum urea and creatinine, blood glucose level and serum amylase. The pancreas was dissected and prepared for histopathological examination by light and electron microscope. Statistical analysis of all collected results was performed. Our biochemical results revealed deleterious effects of renal I-R on the pancreas as evidenced by a highly significant increase in serum amylase and blood glucose level. In I-R group, histopathological examination revealed wide septa and congested blood vessels, acinar cells showed disrupted rough endoplasmic reticulum and few secretory granules. Some islet cells showed pyknotic nuclei and vacuolated cytoplasm. PRP treated group showed nearly normal structure in the form of numerous acinar cells' granules, extensive rough endoplasmic reticulum and mitochondria. Most of Beta cells had euchromatic nuclei and numerous secretory granules. Accordingly, PRP treatment reduced the pancreatic biochemical and histopathological alterations caused by renal I-R injury and so considered a promising therapy for pancreatic damage.

Can pregnancy aggravate the criticality of COVID-19 infection in obese asthmatic peripartum woman? A peripartum COVID-19 mortality case report.

As coronavirus disease 2019 (COVID-19) sweeps all over the world, Information about COVID-19 is evolving rapidly and interim guidance by multiple organisations is constantly being updated and expanded. Early with discovery of COVID 19, it was reported that pregnancy did affect the progress of the disease severity. Recently, Centres for Disease Control and Prevention (CDC) reported that pregnancy is a risk factors for COVID-19 severity. The current case report is presenting a peripartum COVID-19 positive mortality case.

Prasugrel anti-ischemic effect in rats: Modulation of hippocampal SUMO2/3-IкBα/Ubc9 and SIRT-1/miR-22 trajectories.

The beneficial role of prasugrel, a P2Y12 receptor blocker, in several neurointerventional procedures has been reviewed clinically. Beyond its antiplatelet capacity, the potential neuroprotective mechanisms of prasugrel are poorly addressed experimentally. Relevant to the imbalance between neuro-inflammation and neuroprotective pathways in cerebral ischemia/reperfusion (I/R), our study evaluated the anti-ischemic potential of prasugrel treatment through tackling novel targets. Male Wistar rats were allocated into 2 sets; set 1 (I/R 60 min/3 days) to assess the neurological deficits/biochemical impact of prasugrel and set 2 (I/R 60 min/5 days) for evaluating short memory/morphological/immunoreactive changes. Each set comprised 4 groups designated as sham, sham + prasugrel, I/R, and I/R + prasugrel. Post-administration of prasugrel for 3 and 5 days reduced neurological deficit scores and improved the spontaneous activity/short term spatial memory using the Y-maze paradigm. On the molecular level, prasugrel turned off SUMO2/3-inhibitory kappa (Iκ)Bα, Ubc9 and nuclear factor kappa (NF-κ)B. Besides, it inhibited malondialdehyde (MDA) and inactivated astrocytes by downregulating the glial fibrillary acidic protein (GFAP) hippocampal immune-expression. Conversely, it activated its target molecule cAMP, protein kinase (PK)A, and cAMP response element-binding protein (CREB) to enhance the brain-derived nuclear factor (BDNF) hippocampal content. Additionally, cAMP/PKA axis increased the hippocampal content of deacetylator silent information regulator 1 (SIRT1) and the micro RNA (miR)-22 gene expression. The crosstalk between these paths partakes in preserving hippocampal cellularity. Accordingly, prasugrel, regardless inhibiting platelets activity, modulated other cellular components; viz., SUMO2/3-IκBα/Ubc9/NF-κB, cAMP/PKA related trajectories, CREB/BDNF and SIRT1/miR-22 signaling, besides inhibiting GFAP and MDA to signify its anti-ischemic potential.

Virulence attitude estimation of Pasteurella multocida isolates in embryonated chicken eggs.

A total of 220 birds of age ranging from 3 to 14 weeks old were collected from several backyards and different farms in Sharkia Governorate, Egypt, and surveyed for the presence of fowl cholera. Twenty Pasteurella multocida from chickens (15/145, 10%) and ducks (5/75, 6%) were bacteriologically isolated, and it was shown that the infection was significantly related to age and breed. Capsular typing, using multiplex polymerase chain reaction (PCR), demonstrated that all strains were type A (100%). Disk diffusion assay towards ten antimicrobials revealed high susceptibilities to amikacin, doxycycline, chloramphenicol, and neomycin with varying degrees. Doxycycline was effective at the lowest concentration (MIC 0.125-1 µg/ml). Multidrug resistance was detected with a percentage of 25%. Multidrug-resistant isolates (five isolates) were subjected to study their pathogenicity in embryonated chicken eggs (ECE). The results showed a variation in indices between different dilutions of the tested strains. The resulting pathogenicity indices showed significant differences (P < 0.05) according to the origin and dilution of the isolate. From the original inoculum to 10-4 dilutions, the mortality of inoculated embryos occurred within 1-2 days with pathological findings, including maceration and lesions on chorioallantoic membrane (CAM). From dilutions ranging from 10-5 to 10-9, no death occurred until 7 days post-inoculation, but a variation in the lesions on CAM was observed. In conclusion, P. multocida serogroup A could be intensely pathogenic for mature chickens thus causing considerable economic losses, and PCR provides a suitable technique for early and rapid diagnosis of fowl cholera.

Rosuvastatin and low-dose carvedilol combination protects against isoprenaline-induced myocardial infarction in rats: Role of PI3K/Akt/Nrf2/HO-1 signalling.

Rosuvastatin has been shown to activate PI3K/Akt/Nrf2/HO-1 pathway, which promotes cell survival in the myocardium. This study investigated the therapeutic benefit of adding rosuvastatin to low-dose carvedilol in protection against myocardial infarction (MI). Rosuvastatin (RSV) and carvedilol (CAR) were given for 7 consecutive days with concurrent administration of two doses of isoprenaline (ISP) on 6th and 7th days to induce MI. Isoprenaline injections caused detrimental alterations in the myocardial architecture and electrocardiogram (ECG) pattern and significantly increased the infarct size, heart index and serum levels of cardiotoxicity markers compared to the control group. ISP induced oxidative damage, inflammatory and apoptotic events and downregulated PI3K/Akt/Nrf2/HO-1 signalling pathway compared to the control values. Treatment with low-dose CAR and/or RSV prevented the ECG and histopathological alterations induced by ISP, and also reduced the infarct size, heart index, serum creatine kinase-MB, cardiac troponin-I and C-reactive protein levels compared to ISP group. CAR and/or RSV treatment restored the activity of superoxide dismutase and total antioxidant capacity with a consequent reduction in lipid peroxides level. Further, they decreased the expression of nuclear factor (NF)-κB (p65) and increased the phosphorylated PI3K and Akt, which may activate the anti-apoptotic signalling as evidenced by the decreased active caspase 3 level. The combination therapy has a more significant effect in the most studied parameters than their monotherapy, which may be because of the activation of PI3K/Akt Nrf2/HO-1 pro-survival signalling pathway. This study highlights the potential benefits of combining RSV with low-dose CAR in case of MI.

CoQ10 augments candesartan protective effect against tourniquet-induced hind limb ischemia-reperfusion: Involvement of non-classical RAS and ROS pathways.

Tourniquet is a well-established model of hind limb ischemia-reperfusion (HLI/R) in rats. Nevertheless, measures should be taken to alleviate the expected injury from ischemia/ reperfusion (I/R). In the present study, 30 adult male Sprague-Dawley rats were randomly divided into 5 groups (n = 6): control, HLI/R, HLI/R given candesartan (1 mg/kg, P.O); HLI/R given Coenzyme Q10 (CoQ10) (10 mg/kg, P.O); HLI/R given candesartan (0.5 mg/kg) and CoQ10 (5 mg/kg). The drugs were administered for 7 days starting one hour after reperfusion. Candesartan and CoQ10 as well as their combination suppressed gastrocnemius content of angiotensin II while they raised angiotensin-converting enzyme 2 (ACE2) activity, angiotensin (1-7) expression, and Mas receptor mRNA level. Consequently, candesartan and/or CoQ10 reversed the oxidative stress and inflammatory changes that occurred following HLI/R as demonstrated by the rise of SOD activity and the decline of MDA, TNF-α, and IL-6 skeletal muscle content. Additionally, candesartan and/or CoQ10 diminished gastrocnemius active caspase-3 level and phospho-p38 MAPK protein expression. Our study proved that CoQ10 enhanced the beneficial effect of candesartan in a model of tourniquet-induced HLI/R by affecting classical and non-classical renin-angiotensin system (RAS) pathway. To our knowledge, this is the first study showing the impact of CoQ10 on skeletal muscle RAS in rats.

Validation of Camel's Fetal Fluids as Antimicrobial Agents.

The present work investigated the effect of camel's fetal fluids on a variety of bacterial and fungal pathogens. Ten samples of camel's amniotic and allantoic fluids were collected aseptically during parturition and their antimicrobial activities were evaluated by disc diffusion method and minimum inhibitory concentration (MIC) assay. The majority of tested pathogens were inhibited by both fluids up to 25% concentration. The fluids showed zones of inhibition ranging from 8 to 30 mm. The most pronounced inhibition was detected for Staphylococcus aureus, Listeria monocytogenes, Klebsiella pneumonia, and Aspergillus niger but the weak inhibition was obtained for Enterococcus faecalis, Bacillus subtilis, and Candida albicans. Also, the MIC values of amniotic fluids (0.25-2 µg/ml) against Gram-positive bacteria and yeast were lower than the values of allantoic fluids (2-8 µg/ml). But in Gram-negative bacteria and molds, the MIC values of allantoic fluids (0.5-2 µg/ml) were the lowermost. Mucor circinelloides was the only pathogen that resisted both fluids. Analysis of fluid samples revealed the presence of several factors that are known to act as antimicrobial. All tested camel's fetal fluids harbored immunoglobulins, complements, and transferrin. Lysozyme was present in only one of 10 examined samples. We firstly report the prevalence of a profound in-vitro antimicrobial activity of camel's fetal fluids. This activity encourages their use as therapeutic alternative agents to overcome multidrug resistance problems.

Antimicrobial Effect of Propolis Administered through Two Different Vehicles in High Caries Risk Children: A Randomized Clinical Trial.

OBJECTIVE: To investigate the effect of two methods of propolis administration on plaque accumulation and microbial count as well as patient acceptance of each vehicle. STUDY DESIGN: A randomized clinical trial with two parallel arms was used with a sample of 60 high caries risk children 6-8 years old. Children were divided randomly into two groups. Group I: Children who received propolis chewing gum and instructed to chew it twice daily for at least twenty minutes, for two weeks. Group II: children who received propolis mouthwash and instructed to rinse twice daily for one minute. A plaque index was recorded and a plaque sample was collected from all participants at base line and after two weeks of treatment. All participants were asked to rate the preparation they received during treatment period on a Visual Analogue Scale chart. RESULTS: Data showed that propolis had a significant effect on reducing plaque scores and colony counts in both vehicles. There was no significant difference between both vehicles neither on plaque reduction nor on microbial count. However children preferred the gum formula. CONCLUSION: Propolis in both vehicles reduced plaque accumulation and microbial count which recommends its use as an antimicrobial agent in different vehicles.

Ambroxol attenuates cisplatin-induced hepatotoxicity and nephrotoxicity via inhibition of p-JNK/p-ERK.

Hepatotoxicity and nephrotoxicity are important drawbacks of cisplatin. The objective of this study is to evaluate the ability of ambroxol in 2 different doses (35 and 70 mg/kg, i.p.) to protect liver and kidney from damage induced by a single dose of cisplatin (10 mg/kg, i.p.) in comparison with N-acetylcysteine (250 mg/kg, i.p.). Inflammatory, oxidative stress, and apoptotic biomarkers were investigated to show the influence of ambroxol on hepatotoxicity and nephrotoxicity. Ambroxol decreased the elevated activity of liver enzymes (aspartate aminotransferase and alanine aminotransferase) and kidney function tests (blood urea nitrogen and creatinine). Ambroxol mitigated cisplatin inflammatory damage by inhibition of tumor necrosis factor-α, interleukin-1ß, and nuclear factor kappa-B and elevation of nuclear factor erythroid 2-related factor 2. Moreover, ambroxol inhibited oxidative damage indicated by reduction of malondialdehyde and replenished the store of reduced glutathione likely by upregulating glutathione reductase and superoxide dismutase. Elevation of phosphorylated c-Jun N-terminal kinases (p-JNK) and phosphorylated extracellular signal-regulated kinase (p-ERK) were attenuated by ambroxol associated with a decrease in the expression of caspase-3; these results were consistent with histopathological results. These results recommend ambroxol to be co-administered with cisplatin in cancer patients to ameliorate liver and kidney damage, and this was confirmed by MTT assay.

Involvement of Nrf2/HO-1 antioxidant signaling and NF-κB inflammatory response in the potential protective effects of vincamine against methotrexate-induced nephrotoxicity in rats: cross talk between nephrotoxicity and neurotoxicity.

Methotrexate (MTX) is a cytotoxic chemotherapeutic agent widely used in the treatment of cancer and autoimmune diseases like rheumatoid arthritis. However, its use has been limited by its nephrotoxicity. MTX-induced renal injury results in uremia which may influence both the peripheral and central nervous systems causing cognitive and memory problems. The nephroprotective and neuroprotective activities of vincamine (10, 20 and 40 mg/kg), a natural alkaloid with known anti-oxidant, anti-apoptotic and neuroprotective properties, were investigated against MTX-induced toxicity. MTX treatment increased the markers of kidney injury and relative kidney weight, lipid peroxidation, nuclear factor-κB (NF-κB), inflammatory markers, tumor necrosis factor-α, interleukin-1ß, myeloperoxidase and cyclooxygenase-2 and caspase-3 expressions, decreased catalase and superoxide dismutase activities, interleukin-10 and ATP levels and antioxidant proteins, nuclear factor erythroid 2-related factor 2 (Nrf2) and hemeoxygenase-1 (HO-1). Moreover, it disturbed rats' behavior in the locomotor activity test, Y-maze and passive avoidance task. Treatment with vincamine (40 mg/kg) effectively ameliorated MTX-induced renal injury via increasing the expression of Nrf2 and HO-1 suppressing oxidative stress, decreasing the expression of inflammatory markers, NF-κB and caspase-3 pathways and enhancing ATP levels. Additionally, it restored locomotor activity in the locomotor test and memory functions in passive avoidance and Y-maze tests.

Antimicrobial Effect of Garlic (Allium sativum) and Thyme (Zataria multiflora Boiss) Extracts on Some Food Borne Pathogens and Their Effect on Virulence Gene Expression.

The incrementing scope of pathogenic resistance to antibiotics has encouraged the search for antivirulence natural extracts. Therefore, our study designed to demonstrate the antimicrobial activity of an aqueous-garlic and thyme oil extracts against Gram-positive (Staphylococcus aureus) and Gram-negative (Salmonella spp.) bacteria by evaluating the influence of sub-inhibitory concentrations on the expression of the most critical virulence genes of the tested isolates. The antibacterial potential of both herbs was checked by the agar well diffusion method and minimum inhibitory concentration (MIC) assay. Interestingly, all isolates were inhibited by both extracts up to 50% concentration. Also, the MIC values of garlic extract (0.125-1µg/ml) against Salmonella isolates were lower than the values of thyme extract (0.5- 8µg/ml). But in S. aureus isolates, the MIC values of thyme extract (0.25- 2µg/ml) were the lowermost. Conventional PCR investigated that all S. aureus isolates carried the hlg (hemolysin) and icaA (intracellular adhesion) genes, but only six Salmonella isolates (three S. typhimurium and one each of S. kentucky, S. anatum, and S. lagos) had both the sopB (Salmonella outer protein B) and mgtC (membrane protein) genes. Real-time RT-PCR assays were performed to evaluate the extract's effect on the virulence genes. The thyme-oil extract has significantly repressed S. aureus virulence genes expression more than aqueous-garlic extract, which later one has effectively more than thyme-oil extract in downregulating the Salmonella virulence genes. In conclusion, garlic and thyme extracts can be used not only as a flavor, but also as potential antimicrobial agents against Gram-positive and negative bacteria.