Repaglinide-Solid Lipid Nanoparticles in Chitosan Patches for Transdermal Application: Box-Behnken Design, Characterization, and In Vivo Evaluation.

Background: Repaglinide (REP) is an antidiabetic drug with limited oral bioavailability attributable to its low solubility and considerable first-pass hepatic breakdown. This study aimed to develop a biodegradable chitosan-based system loaded with REP-solid lipid nanoparticles (REP-SLNs) for controlled release and bioavailability enhancement via transdermal delivery. Methods: REP-SLNs were fabricated by ultrasonic hot-melt emulsification. A Box-Behnken design (BBD) was employed to explore and optimize the impacts of processing variables (lipid content, surfactant concentration, and sonication amplitude) on particle size (PS), and entrapment efficiency (EE). The optimized REP-SLN formulation was then incorporated within a chitosan solution to develop a transdermal delivery system (REP-SLN-TDDS) and evaluated for physicochemical properties, drug release, and ex vivo permeation profiles. Pharmacokinetic and pharmacodynamic characteristics were assessed using experimental rats. Results: The optimized REP-SLNs had a PS of 249±9.8 nm and EE of 78%±2.3%. The developed REP-SLN-TDDS demonstrated acceptable characteristics without significant aggregation of REP-SLNs throughout the casting and drying processes. The REP-SLN-TDDS exhibited a biphasic release pattern, where around 36% of the drug load was released during the first 2 h, then the drug release was sustained at around 80% at 24 h. The computed flux across rat skin for the REP-SLN-TDDS was 2.481±0.22 µg/cm2/h in comparison to 0.696±0.07 µg/cm2/h for the unprocessed REP, with an enhancement ratio of 3.56. The REP-SLN-TDDS was capable of sustaining greater REP plasma levels over a 24 h period (p<0.05). The REP-SLN-TDDS also reduced blood glucose levels compared to unprocessed REP and commercial tablets (p<0.05) in experimental rats. Conclusion: Our REP-SLN-TDDS can be considered an efficient therapeutic option for REP administration.

Genome sequencing of Enterococcus faecium NT04, an oral microbiota revealed the production of enterocin A/B active against oral pathogens.

Objective: This study aimed to isolate and investigate a bacterium from an Egyptian adult's healthy oral cavity, focusing on its probiotic properties, especially its antagonistic activity against oral pathogens. Methods: The isolated bacterium NT04 using 16S rRNA gene sequencing, was identified as Enterococcus faecium. In this study, the whole genome of Enterococcus faecium NT04 was sequenced and annotated by bioinformatics analysis tools. Results: Numerous genes encoding the production of diverse metabolic and probiotic properties, such as bacteriocin-like inhibitory substances (Enterocin A and B), cofactors, antioxidants, and vitamins, were confirmed by genomic analysis. There were no pathogenicity islands or plasmid insertions found. This strain is virulent for host colonization rather than invasion. Conclusion: Genomic characteristics of strain NT04 support its potentiality as an anti-oral pathogen probiotic candidate.

The mitigating effect of Spirulina (Arthrospira platensis) on the hemotoxicity of gibberellic acid on juvenile tilapia (Oreochromis niloticus).

The use of plant growth regulators has led to environmental contamination of water bodies that occur adjacent to agricultural areas. Some of these chemicals are bioactive, not only to plants, but also to non-target exposed biota, namely of the aquatic compartment. Previous work demonstrated the establishment of hepato- and nephrotoxic effects in juvenile tilapia (Oreochromis niloticus) exposed via aquatic media to gibberellic acid (GA3), which is among the most used plant growth regulators, in agricultural practices. Here, we investigated the effect of GA3 on hematological indices, poikilocytosis, nuclear abnormalities, and genotoxic indices measured in Nile tilapia (Oreochromis niloticus), as well as the putative protective effects of dietary supplementation of Spirulina (Arthrospira platensis). Fish were evenly assorted into 5 groups: group I served as a control, and groups II-V were fed diets supplemented with Spirulina at rates of 0 g/kg, 5 g/kg, 20 g/kg, and 100 g/kg, respectively, for 2 months before being exposed to 150 mg/L GA3. The results revealed that GA3 exposure decreased significantly all hematological indices (P < 0.05), except leucocytes and mean corpuscular hemoglobin concentration (MCHC), compared to the control group (P > 0.05). GA3 exposure increased significantly the percentage of nuclear abnormalities, altered erythrocytes and the percentages of tail DNA, compared to the control group (P < 0.05). Spirulina supplementation restored the hematological, poikilocytosis, nuclear abnormalities, and the percentages of tail DNA to near normal levels. The 100 g/kg SP treatment was the most effective in attaining such effect, showing concentration-dependency. The present study reinforces our findings of the toxicity of GA3 on O. niloticus and suggests that the addition of Spirulina to fish diet can mitigate the hemotoxic effects of GA3.

Biosimilars according to Jordan Food and Drug Administration regulations.

In 2015, JFDA approved its biosimilars registration guidelines [1] officially. Many steps have been taken before achieving this progress. This paper summarizes briefly JFDA efforts that were done in the previous years and the actions taken to approve JFDA biosimilars registration guidance which was based on EMA related guidelines [2-5], and its impact on enhancing the affordability of safe, effective and high quality biosimilars for patients.

Potent synergistic combination of rosuvastatin and levofloxacin against Staphylococcus aureus: in vitro and in vivo study.

AIMS: The present study aims to evaluate the capability of rosuvastatin to synergize with levofloxacin against Staphylococcus aureus. METHODS AND RESULTS: Rosuvastatin inhibited the growth of S. aureus with minimum inhibitory concentration of 16 µg ml-1 . Additionally, it showed a bactericidal effect at 4x minimum inhibition concentration. Using a checkerboard method, a synergistic effect was recorded when rosuvastatin was combined with levofloxacin showing against S. aureus isolate 28 (S 28). Furthermore, this combination was also able to display a significant reduction in biofilm formation (92·8%) and suppress the production of coagulase and ß-haemolysin, and virulence factors of S. aureus isolate 28. An animal model for wound infection was used to assess the therapeutic effect of the test combination, in vivo. It was found that the test combination reduced the bacterial burden in the infected wounds by 91·3%. Pathological and histological analyses have revealed a decline in cell infiltration in the excisional wound skin tissue after treatment with rosuvastatin and levofloxacin combination. CONCLUSIONS: Rosuvastatin combined with levofloxacin can be considered as a promising solution to combat S. aureus antibiotic resistance phenomenon. SIGNIFICANCE AND IMPACT OF THE STUDY: This study unveils the potential effect of rosuvastatin when used in combination with levofloxacin can be used as a topical antibacterial agent to treat S. aureus skin infections.

Lung ultrasound versus chest radiography for the diagnosis of pneumothorax in critically ill patients: A prospective, single-blind study.

BACKGROUND: Radiologic data remains the gold standard for the diagnosis of pneumothorax (PTX). The use of ultrasonography (US) has recently emerged as the method of choice with physicians who can perform bedside US. PURPOSE: To compare the diagnostic accuracy of lung US against bedside chest radiography (CR) for the detection of PTX using thoracic computed tomography (CT) as the gold standard. MATERIALS AND METHODS: We conducted a prospective, single-blind study on 192 critically ill patients; each patient received lung US examination, bedside CR, followed by thoracic CT scan searching for PTX. RESULTS: Of the studied patients, CT of the chest confirmed the diagnosis of PTX in 36 (18.75%) patients of which 31 were diagnosed by thoracic US while CR detected only 19 cases. Overall lung US showed a considerable higher sensitivity than bedside CR (86.1% vs. 52.7%), lung US also showed higher, negative predictive values, and diagnostic accuracy against CR (96.8% vs. 90.1%), and (95.3% vs. 90.6%), respectively. CR had a slightly higher specificity than lung US (99.4% vs. 97.4%), and higher positive predictive values (95.0% vs. 88.6%). CONCLUSION: Lung US is an accurate modality more than anteroposterior bedside CR in comparison with CT scanning when evaluating critically ill mechanically ventilated patients, patients underwent thoracocentesis, central venous catheter insertion, or patients with polytrauma.

Pathological findings in equine herpesvirus 9-induced abortion in rats.

Pregnant rats were infected experimentally with equine herpesvirus (EHV)-9, a new neurotropic equine herpesvirus serologically similar to EHV-1, during the first and third trimesters. The inoculated dams had mild to severe neurological signs and gave birth to dead fetuses or undersized pups. Rats inoculated during the first and last trimesters had varying degrees of encephalitis as well as abnormalities of the placentas in the form of marked dilation of maternal blood sinusoids and varying degrees of atrophy and necrosis of the trophoblast cells of the labyrinth, the spongiotrophoblasts and the giant cell layer. Virus antigen was detected by immunohistochemistry in the brain and the trophoblast cells of labyrinth, the spongiotrophoblasts and giant cell layer of the placenta in rats inoculated during the first trimester. Virus antigen was detected in fetuses from rats inoculated in the first and last trimesters. Virus DNA was amplified by polymerase chain reaction from the placenta and fetuses of inoculated rats. EHV-9 may induce fetal death and abortion in pregnant dams, possibly caused by direct EHV-9 infection of the placenta and/or fetus as well as the secondary effect of vascular injury.

Kinetics and pathogenicity of oral infection by equine herpesvirus-9 in mice and suckling hamsters.

The pathogenesis and kinetics of oral infection by equine herpesvirus (EHV)-9 were studied in mice and hamsters. After oral inoculation of 10(5) plaque-forming units (PFU) of virus, 1-week-old suckling hamsters showed varying severity of neurological disease from 72 hours post inoculation (hpi) and all of these animals had died by 96 hpi. Four-week-old ICR mice inoculated orally with 4 × 10(4)PFU of virus showed no clinical signs, but they developed erosive and ulcerative gastritis from 36 hpi. Varying degrees of encephalitis were seen in infected mice and hamsters, and the hamsters also developed myelitis by 96 hpi. Immunohistochemistry performed on whole body sections of suckling hamsters revealed the kinetics of spread of the virus to the central nervous system. EHV-9 antigen was detected initially in macrophages of the oral and lingual submucosa. At 36 hpi virus antigen was detected in the nerve fibres and pseudounipolar neurons of the trigeminal ganglion and at 96 hpi antigen was present in the myenteric plexuses of the intestine. Virus antigen was also detected in the liver, lungs and heart of affected animals. EHV-9 DNA was detected by polymerase chain reaction in the brain, blood and spinal cord of suckling hamsters at 36, 48 and 96 hpi. These findings show that EHV-9 may spread via the trigeminal nerve when mice and hamsters are inoculated orally with virus.

The effect of oral activated charcoal on the systemic clearance of gentamicin in rabbits with acute renal failure.

The effect of oral administration of activated charcoal on total body clearance of gentamicin administered intravenously (2 mg kg-1) has been studied in normal rabbits and rabbits with induced renal failure. Gastric intubation of a single dose (10 g) of activated charcoal to normal rabbits produced a significant reduction in gentamicin serum concentrations compared to control. Significant differences between treated and control groups, compatible with enhancement of gentamicin elimination, were observed in the calculated pharmacokinetic parameters (Kel, t 1/2, CL and AUC). To examine whether renal failure could augment the effect of activated charcoal in enhancing the systemic clearance of gentamicin, uranyl nitrate was used (0.75 mg kg-1, i.v.) to induce acute renal failure in rabbits. The derived pharmacokinetic parameters of gentamicin during the control phase in these animals were consistent with severe renal failure. The administration of activated charcoal, 2.5 h following gentamicin injection, produced a steeper decline in gentamicin concentration-time profiles and significant changes in Kel, t 1/2 and CL. The Kel and CL values increased to about 200%, while the t 1/2 value decreased to about 50%. The apparent changes in the pharmacokinetic parameters induced by charcoal administration were more marked in rabbits with renal failure than in normal rabbits; however, induction of renal failure did not augment the charcoal-induced clearance of gentamicin quantitatively.

Cytotoxicity and mutagenicity of 'Al-Kohl', an eye cosmetic commonly used in Jordan.

We examined 50 samples of 'Al-Kohl', collected from northern Jordanian provinces, for their cytotoxicity and mutagenicity using the brine shrimp and Ames Salmonella mutagenicity bioassays, respectively. Twenty were unopened, ready-to-use, samples purchased from retail outlets, 20 were in-use samples obtained from ladies of different socioeconomic standards, and 10 samples were from the original stones used to prepare Al-Kohl. The frequency of positive samples for both cytotoxicity and mutagenicity was much higher in the ready-to-use and in-use samples of Al-Kohl than in the original stones. Out of the 50 samples examined, 20 (40%) showed some degree of cytotoxicity almost all involving ready-to-use or in-use samples. Among those samples, 12 (24%) were strongly cytotoxic, while eight samples (16%) showed mild cytotoxic activity in the brine shrimp bioassay. The results of the mutagenicity testing were obtained, without using any metabolic activation, with four test strains of Salmonella, namely TA97, TA98, TA100 and TA102. Wide variability in results was observed depending on the type of samples tested and the Salmonella strain used. More than 80% of the original stone samples were negative and the positive ones were mildly mutagenic while the ready-to-use and in-use samples showed similar mutagenicity, which was much more than the original stones against the four strains of Salmonella typhimurium used. Strain TA97 was particularly sensitive to the samples tested. Twelve per cent of the ready-to-use and in-use samples of Al-Kohl showed a strong mutagenic effect against the pre-mentioned strains. We recommended abandoning Al-Kohl as a cosmetic for application to the eye areas, based on these findings and the previous microbial contamination studies (2).

Microbial contamination of cosmetics and personal care items in Egypt--body lotions and talcum powders.

We examined a total of 54 samples, including 18 body lotions and 36 talcum powders, for their total aerobic bacterial, coliform and fungal counts. We also carried out anaerobic bacterial counts for talcum powder as well as tests to detect some potentially hazardous bacteria in all tested samples. Talcum powders were more heavily contaminated with bacteria and fungi than body lotions. More than 40% of the tested body lotions contained no viable bacteria or less than 100 c.f.u./g. while all the talcum powders tested contained more than 100 c.f.u./g. Thirty per cent of the talcum powders were contaminated with 10(4) c.f.u./g. and none of the body lotions were contaminated to that extent. No coliforms were recovered from any of the body lotions, while 17% of the talcum powder examined contained coliforms in the range of 230-500 c.f.u./g. Staphylococcus spp. were detected in 18 samples of both talcum powders and body lotions, three of these Staphylococci were of the aureus type. Three samples of talcum powder contained E. coli, two samples contained Enterobacter agglomerans and one sample contained Citrobacter freundii. Seventy per cent of the body lotions showed no fungal counts, while 83% of the talcum powders examined were contaminated with fungi and most of the contaminated talcum powders contained more than 100 fungal cells/g. With regard to the anaerobic bacterial counts for talcum powders, 50% of the samples showed no counts while the other 50% contained less than 100 c.f.u./g. Four samples were contaminated with Clostridium perfringens, although C. tetani was not recovered from any of the samples.

Microbiological profile of selected samples of "Al-Kohl" eye cosmetics in northern Jordanian provinces before and after use.

Fifty items of Al-Kohl collected from northern Jordanian provinces representing: 20 unopened ready to use samples purchased from retail outlets, 20 in-use samples obtained from ladies of different socioeconomic standards and 10 samples of the original stones (used for Al-Kohl preparation) were examined for their microbial contents. Ready to use and in-use samples were much more contaminated than the original stones. On sterility testing, more than 85% of the unused and in-use samples were contaminated with bacteria and or fungi comparing to 50% of the original stone samples. Quantitatively, 90% of the original stones contained less than 100 bacterial or fungal cells/g and the other remaining 10% were in the range of 10(2)-10(3) cfu/g either for bacterial or fungal counts. The level and distribution of the viable microbial counts in unused and in-use samples were comparable and much higher than original stones. More than 70% and 20% of those items contained more than 100 cfu/g of bacteria and fungi respectively. Among those samples 20% and 5% were heavily contaminated (contain more than 10(4) cfu/g) with bacteria and fungi respectively. Coliform bacteria in a number of 100 cfu/g or more were recovered from 10% of the unused and 20% of the in-use samples, none were recovered from original stones. The results of qualitative tests for identification of isolated microorganisms showed that 7 different species of Bacillus were found in the 50 examined samples. Approximately 50% of the examined samples contained Bacillus spp., Staphylococcus spp., Pseudomonas spp., P. vulgaris, S. marcescens were recovered from unused and in-use samples in different percentages, none from original stones, some of the detected Staphylococcus were aureus type and also one isolate of P. aeruginosa was detected in one of the in-use samples. The relationship between the detected level of microbial contamination in the tested samples with the proposed allowable international limits of contamination as well as the possible sources of contamination and the hygienic implications of using such products by the public were discussed.

Microbial contamination of cosmetics and personal care items in Egypt--eye shadows, mascaras and face creams.

We examined a total of 150 samples, including 27 eye shadows, 27 mascaras and 96 face creams, for their microbial contents. Mascaras were generally more contaminated than eye shadows. More than 75% of the examined eye shadows contained fewer than 100 c.f.u./g aerobic bacterial count compared to 63% of the mascaras examined. Viable bacteria were not recovered from 61% and 48% of the eye shadows and mascaras respectively. While 4% of the eye shadows were heavily contaminated (contained more than 10(4) c.f.u./g), 15% of the mascaras were as heavily contaminated (with more than 10(4) c.f.u./ml of bacteria). Face creams were generally more heavily contaminated than eye shadows and mascaras. More than 70% of the examined creams contained more than 100 c.f.u./g of bacteria compared to 23% and 37% of eye shadows and mascaras respectively. Only 5% of the face creams were heavily contaminated. However, 27% of the creams were contaminated with more than 10(3)-10(4) c.f.u./g of bacteria compared to none in this range for both eye shadows and mascaras. Qualitative tests for detection of hazardous bacteria showed that none of the eye shadows were contaminated with any of those micro-organisms. Out of nine items of a specific brand of mascara, three isolates of Pseudomonas aeruginosa, one isolate of Citrobacter freundii and one isolate of Klebsiella pneumonia were detected. Among the creams, two brands showed the highest contamination levels with more than 85% of the tested samples containing more than 10(3) c.f.u./g fungi and at least 10(4) c.f.u./g bacteria.(ABSTRACT TRUNCATED AT 250 WORDS)

Microbial contamination of cosmetics and personal care items in Egypt--shaving creams and shampoos.

We examined a total of 192 samples, including eight different brands of shaving cream and eight brands of shampoo, for their total aerobic bacterial, coliforms and fungal counts. Shaving creams were more heavily contaminated with bacteria than shampoos. Viable bacterial were not recovered from 57% and 10% of shampoos and shaving creams, respectively. Only 3% of shaving creams were heavily contaminated with more than 10(4) c.f.u./g, while none of the shampoos contained such a high number of bacteria. With regard to the medium range contamination levels, 52% of shaving creams showed bacterial counts ranging from 10(2) to 10(3) c.f.u./g or ml, compared to 15% of shampoos which were contaminated to the same level. Fourteen per cent of shaving creams were contaminated with greater than 10(3)-10(4) c.f.u./g or ml, compared to 1% of the shampoos. No coliforms were recovered from either the shaving creams or the shampoos; however, Staphylococcus spp. were detected in six samples of both shampoos and shaving creams. Some of these Staphylococci, were aureus type. One isolate of Pseudomonas aeruginosa was also detected in a sample of shampoo. The incidence of fungal contamination was much less than the bacterial contamination. No viable fungi were recovered from 88% and 76% of the shaving creams and shampoos, respectively. The majority of the remaining samples, for both products, were contaminated with less than 100 fungal cell/g or ml. The pH of all the tested samples was alkaline (pH 7.2-9), which is well known to inhibit fungal contamination.

Sporicidal activity of local anaesthetics and their binary combinations with preservatives.

The sporicidal activity of a 1% solution of five local anaesthetics and five preservatives (cetrimide, chlorocresol, chlorhexidine, phenoxyethanol and phenylmercuric nitrate) at their commonly used concentrations, alone and in binary combinations, was determined against Bacillus subtilis and Aspergillus niger spores at different temperature levels by surface viable count technique. The sporicidal activity of all tested systems was temperature dependent and A. niger spores were much more sensitive to the effect of the test systems than B. subtilis spores. The temperatures at which 99% kill is achieved after 30 min exposure were calculated. For local anaesthetics used singly against A. niger the recorded temperatures were 30 degrees C for amethocaine, 45 degrees C for amylocaine, 43 degrees C for cincochaine, 48 degrees C and 50 degrees C for lignocaine and procaine, respectively. A control temperature of 58 degrees C for saline solution was observed. Much higher temperatures were recorded for B. subtilis spores. Cincochaine was the most effective local anaesthetic with a recorded temperature of 60 degrees C for a 99% kill while amylocaine and amethocaine showed temperatures of 84 and 90 degrees C respectively. Procaine, lignocaine as well as the control saline solution recorded temperatures higher than 100 degrees C. Among the 25 binary combinations of local anaesthetics and preservatives tested, the most pronounced potentiation of the sporicidal activity against fungal spores was recorded with chlorocresol combinations, while other combinations of the four remaining preservatives showed different types of interactions at various temperature levels.

Microbial contamination of a hexetidine mouth wash.

Twenty different bottles of a mouth wash containing 0.05% hexetidine as an active ingredient were examined for their microbiral contents. The results showed that all the tested bottles were contaminated with bacteria. Nine out of the twenty bottles contained more than 10(4) CFU/ml. Staphylococcus aureus was detected in one bottle, while Pseudomonas species were found in four bottles. Fungi were detected in 10 bottles, a fungal count of more than 100 fungus/ml were found in 12 out of the 19. Yeasts were detected in 16 bottles, Candida species were the most predominant with a rate of 11 out of the 16, while Saccharomyces species were found in 5 out of the 16. C. albicans, a definite oral pathogen, was found in 3 bottles.

Sporicidal activity of local anaesthetics and their binary combinations with preservatives.

The sporicidal activity of 1% solutions of five local anaesthetics and five preservatives (cetrimide, chlorocresol, chlorhexidine, phenoxyethanol and phenylmercuric nitrate) at their pharmacopeial concentrations, alone and in binary combinations was determined against Bacillus subtilis and Aspergillus niger spores at different temperature levels by the surface viable count technique. The sporicidal activity of all tested systems were temperature dependent. A. niger spores were much more sensitive to the tested systems than B. subtilis spores. The temperature at which 99% of all spores were killed after 30 min exposure time of the anaesthetics were calculated. In case of the local anaesthetics alone against spores of A. niger the recorded temperatures were 30 degrees C for amethocaine, 45 degrees C for both amylocaine and cincochaine, 48 degrees C and 50 degrees C for lignocaine and procaine, respectively, in contrast to 58 degrees C in a control with saline solution. Much higher temperatures were calculated against B. subtilis spores. Cincochaine was the most effective local anaesthetic with a recorded temperature of 60 degrees C, where 99% killing occurred. Amylocaine and amethocaine showed temperatures of 84 degrees C and 90 degrees C, respectively. Procaine, lignocaine as well saline solution as a control caused a 99% killing effect at temperatures higher than 100 degrees C. Among the 25 tested binary combinations of local anaesthetics and preservatives, the highest incidence of potentiation of the sporicidal activity was recorded with chlorocresol combinations, while other combinations of the four remaining preservatives showed different types of interactions at different percentages.