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Background Green fluorescent protein (GFP) is a valuable macromolecule widely utilized in biomedical diagnostics and diverse biotechnological applications due to its low toxicity and intrinsic fluorescence, which does not require exogenous substrates or cofactors. Nevertheless, the conventional Aequorea victoria –derived GFP exhibits limitations, including relatively low brightness and restricted applicability under low-oxygen conditions. Consequently, there is increasing interest in identifying alternative and less explored microbial sources of GFP, particularly from marine environment. Results In the current study, eleven fluorescent bacteria were isolated from different anatomical parts of squid collected from Miami Beach (Alexandria, Egypt) and from seawater sample from Atubia island (Safaga, Egypt). Among the three isolation media evaluated, Sea Water Complete agar medium (SWC) supported the highest recovery of fluorescent bacteria. Fluorescence-based primary and secondary screening identified isolate E2 as the most efficient GFP producer. It was identified phenotypically using the VITEK ® 2 automated system and genotypically using 16 S rRNA studies as Pseudomonas gessardii with 99.34% similarity. Sequences of the 16 S rRNA gene were deposited in the GenBank as OQ285875. Optimization of environmental condition using a one-factor-at-a-time approach revealed the maximum GFP production was achieved after 30 h of incubation at 25 °C and pH 7.5 under static conditions (0 rpm). These optimized parameter enhanced the fluorescence intensity of GFP by 1.51-fold compared to the basal medium. Subsequent purification using an aqueous two-phase system and fast protein liquid chromatography successfully recovered GFP with a high purity of 96.3%. Conclusion Overall, the current study identifies Pseudomonas gessardii E2 strain as a novel marine-derived source of high-quality pure GFP and establishes an effective strategy for its production and purification. The findings provide a promising foundation for the potential application of this GFP in bioimaging, biosensing, and fluorescence-based reporter systems, including applications in oxygen-limited environments.

Apocynin (APO) is a bioactive phytochemical with prominent anti-inflammatory and anti-oxidant activities. Designing a nano-delivery system targeted to potentiate the gastric antiulcerogenic activity of APO has not been investigated yet. Chitosan oligosaccharide (COS) is a low molecular weight chitosan and its oral nanoparticulate system for potentiating the antiulcerogenic activity of the loaded APO has been described here.  COS-nanoparticles (NP ) loaded with APO (using tripolyphosphate [TPP] as a cross-linker) were prepared by ionic gelation method and fully characterized. The chosen formula was extensively evaluated regarding in vitro release profile, kinetic analysis, and stability at refrigerated and room temperatures. Ultimately, the in vivo antiulcerogenic activity against ketoprofen (KP)-induced gastric ulceration in rats was assessed by macroscopic parameters including Paul's index and antiulcerogenic activity, histopathological examination, immunohistochemical (IHC) evaluation of cyclooxygenase-2 (COX-2) expression level in ulcerated gastric tissue, and biochemical measurement of oxidative stress markers and nitric oxide (NO) levels. The selected NP formula with COS (0.5 % w/v) and TPP (0.1% w/v) was the most appropriate one with drug entrapment efficiency percentage of 35.06%, particle size of 436.20 nm, zeta potential of +38.20 mV, and mucoadhesive strength of 51.22%. It exhibited a biphasic in vitro release pattern as well as high stability at refrigerated temperature for a 6-month storage period. APO-loaded COS-NP provoked marvelous antiulcerogenic activity against KP-induced gastric ulceration in rats compared with free APO treated group, which was emphasized by histopathological, IHC, and biochemical studies.  In conclusion, APO-loaded COS-NP could be considered as a promising oral phytopharmaceutical nanoparticulate system for management of gastric ulceration.

Recently, Apocynin (APO) has emerged as a bioactive phytochemical with potent antioxidant and anti-inflammatory properties. No reports have been published so far concerning its topical application as a pharmaceutical dosage form for prospective use. To the best of our knowledge, this is the first study to fabricate novel anti-inflammatory film for external medication with APO. APO film was prepared using casein (CAS) as a natural protein film former by solvent casting technique. The medicated film was extensively evaluated in terms of its various physicochemical characteristics, ex vivo skin permeation profile, stability, and finally in vivo anti-inflammatory activity on carrageenan-induced rat paw edema. The film represented satisfactory mechanical properties along with good flexibility. Fourier transform-infrared spectroscopy, differential scanning calorimetry, and X-ray diffractometry revealed possible solubility of APO in the amorphous CAS and inter- and intramolecular hydrogen bonding between the film components. The ex vivo skin permeation results of the medicated film demonstrated non-Fickian diffusion mechanism of the permeated drug. Application of APO film to rat paw before carrageenan-induced paw edema or after induction disclosed eminent anti-inflammatory activity expressed by marked decrease in paw swelling (%) and increase in edema inhibition rate (%). In addition, histopathologic examination revealed a significant decrease in inflammatory scores. The immunohistochemical expression levels of both nuclear factor kappa B and cyclooxygenase-2 were significantly suppressed. These results indicated that CAS film could be applied as a promising external delivery system for the anti-inflammatory APO.

Chemotherapy, particularly cisplatin, is a prevalent cancer treatment. Unfortunately, many tissues, for instance the submandibular salivary glands, are toxically affected by cisplatin. Of significant interest, phytopharmaceuticals rich in flavonoids have demonstrated exceptional defense against chemotherapy induced toxicity, like chrysin (Chr); nevertheless, its low solubility and poor bioavailability have remained cornerstone issues. Accordingly, Chr was successfully encapsulated in the poly(d, l-lactide-co-glycolide) nanoparticles (PLGA NPs) scaffold. The developed chrysin-loaded poly(d, l-lactide-co-glycolide) nanoparticles (Chr-loaded PLGA NPs) were meticulously evaluated via comprehensive in vitro-in vivo investigations. Saliently, the outcomes of in vivo studies exhibited admirable in vivo counteraction effectiveness against cisplatin-induced toxicity in submandibular salivary glands in Albino rats upon comparing Chr-loaded PLGA NPs treated group with pure Chr as well as blank NPs treated ones. Inclusively, Chr-loaded PLGA NPs can be regarded as promising therapy to create recent vistas for dampening myriad adverse effects of different chemotherapies.

Background: Apocynin (APO) is a bioactive phytochemical with prominent anti-inflammatory and anti-oxidant activities. Designing a nano-delivery system targeted to potentiate the gastric antiulcerogenic activity of APO has not been investigated yet. Chitosan oligosaccharide (COS) is a low molecular weight chitosan and its oral nanoparticulate system for potentiating the antiulcerogenic activity of the loaded APO has been described here. Methods: COS-nanoparticles ([NP.sub.s]) loaded with APO (using tripolyphosphate [TPP] as a cross-linker) were prepared by ionic gelation method and fully characterized. The chosen formula was extensively evaluated regarding in vitro release profile, kinetic analysis, and stability at refrigerated and room temperatures. Ultimately, the in vivo antiulcerogenic activity against ketoprofen (KP)-induced gastric ulceration in rats was assessed by macroscopic parameters including Paul's index and antiulcerogenic activity, histopathological examination, immunohistochemical (IHC) evaluation of cyclooxygenase-2 (COX-2) expression level in ulcerated gastric tissue, and biochemical measurement of oxidative stress markers and nitric oxide (NO) levels. Results: The selected [NP.sub.s] formula with COS (0.5 % w/v) and TPP (0.1% w/v) was the most appropriate one with drug entrapment efficiency percentage of 35.06%, particle size of 436.20 nm, zeta potential of +38.20 mV, and mucoadhesive strength of 51.22%. It exhibited a biphasic in vitro release pattern as well as high stability at refrigerated temperature for a 6-month storage period. APO-loaded COS-[NP.sub.s] provoked marvelous antiulcerogenic activity against KP-induced gastric ulceration in rats compared with free APO treated group, which was emphasized by histopathological, IHC, and biochemical studies. Conclusion: In conclusion, APO-loaded COS-[NP.sub.s] could be considered as a promising oral phytopharmaceutical nanoparticulate system for management of gastric ulceration. Keywords: apocynin, chitosan oligosaccharide, tripolyphosphate, nanoparticles, antiulcerogenic activity

Background Studies about perimenopausal migraine are limited. In this study, we aimed to explore the characteristics of perimenopausal migraine and to compare perimenopausal women with and without migraine in terms of the levels of Estradiol and Follicle-Stimulating Hormone (FSH). We also aimed to correlate migraine severity scores with Estradiol and FSH levels. Methods A case-control study of 66 subjects divided as Group 1 ( n  = 22): migraine onset during perimenopause, Group 2 ( n  = 22) migraine onset before perimenopause, and Group 3 ( n  = 22) age-matched healthy controls. Perimenopausal women with any type of migraine (age 40–55 years) were eligible. We excluded secondary and other primary headaches, hormonal therapy and chronic illness. Subjects underwent a detailed history taking of headache and vasomotor symptoms, MIDAS and VAS scores. Measurement of Estradiol (E2) and FSH levels was done for all participants. Comparisons were done between group 1 and group 2 by migraine characteristics, vasomotor symptoms and Estradiol and FSH levels. Group 3 (control) was compared with both migraine groups 1 and 2 by vasomotor symptoms and Estradiol and FSH levels. Statistical analysis was performed by using SPSS 27th edition Statistical tests used include mean, median, count, Mann-Whitney U, Kruskal-Wallis test, Chi-squared test and the Spearman correlation test (The suitable test was chosen based on the type of variables and data, and the number of comparison groups). Results Migraine with onset during perimenopause was characterized by throbbing pain, alternating laterality, orbito-fronto-temporal site, moderate to severe intensity, high frequency, with associated phonophobia and photophobia, while autonomic symptoms were rare. There was analgesic overuse and an unsatisfactory response. Group 2 significantly had more postdrome symptoms, less analgesic response and more menstrually-triggered migraine than Group 1 (P < 0.05). Both perimenopausal migraine groups (Group 1 and 2) had significantly more perimenopausal symptoms, including fatigue, sleep disturbance, mood symptoms and anxiety (P < 0.05) compared to the control (Group 3). Hot flashes, night sweating and vaginal dryness were more common in migraineurs without statistical significance. Migraineurs had significantly lower Estradiol levels and a higher proportion of subjects with Estradiol < 50 pg./ml than controls (P < 0.05). There was a significant negative correlation between Estradiol and scores, including MIDAS (Rho=-0.406, P  = 0.007) and VAS (Rho= -0.331, P  = 0.030). No difference in the FSH levels among the Groups. Conclusion Perimenopause is associated with high-frequency, moderate to severe migraine headache. Lower Estradiol levels were correlated with high migraine severity scores (VAS and MIDAS). The frequency of perimenopausal symptoms is higher, whereas the Estradiol level is lower in migrainous compared to nonmigrainous women.