Novel sprayable thermosensitive benzydamine hydrogels for topical application: Development, characterization, and in vitro biological activities

Benzydamine hydrochloride (BZD) having analgesic, anesthetic, and anti-inflammatory effects is used orally or topically in the treatment of disorders such as joint inflammation and muscle pain. Within the scope of this study, sprayable thermosensitive BZD hydrogels were developed using thermoresponsive poloxamers to avoid systemic side effects and to provide better compliance for topical administration. Also, hydroxypropyl methyl cellulose (HPMC) was employed to improve the mechanical strength and bioadhesive properties of the hydrogel. The addition of BZD generally decreased the viscosity of the formulations (p 0.05); the release profiles of BZD from the formulations were similar according to model-independent kinetic (f2 > 50). HPMC and poloxamers had important roles in the accumulation of BZD in the skin. In vitro biological activity studies demonstrated that the formulations presented their anti-inflammatory activity with TNF-α inhibition but did not have any effect on the inhibition of COX enzymes as expected. As a result, thermosensitive hydrogels containing BZD might be an appropriate alternative, providing an advantage in terms of easier application compared to conventional gels. Graphical Abstract: [Figure not available: see fulltext.]BAS

Determination of Viable Salmonella Typhimurium Cells in Heat Treated Milk By PMA/Real-Time PCR Method

Applying different technological processes during the production of food has a lethal effect on the bacteria but DNA of these bacterial strains may cause false positive results when detected by real time PCR technique because they preserve their existence for a certain period of time. To overcome this shortcoming of the real time PCR technique, a new method has been developed in recent years, based on the removal of dead cell DNA from the medium by treatment with Propodium Monoazide (PMA) before DNA extraction. In this study, real-time PCR method was combined with PMA application for the detection of live cells of Salmonella Typhimurium in heat treated milk samples. For this purpose, milk samples inoculated with S. Tyhimurium were heat treated at different temperatures (60, 65, 70 and 75°C) and times (15, 60, 300, 900 sec) and number of live bacteria was determined comparatively by direct real-time PCR, PMA/real-time PCR and conventional cultural method. As a result, unlike the direct real time PCR technique, PMA/real-time PCR method prevents to a certain extent of false positive results from dead cells at all tested temperatures and times but higher results were obtained from PMA/real-time PCR method when compared to conventional cultural results. Therefore, further studies should be carried out to optimize the conditions of the PMA application in order to eliminate the high positive results detected by the PMA / real-time PCR metho

Tavuk yumurtasından immunoglobulin y (ıgy) eldesi ve etkinliğinin gösterilmesi

1. Dünya Sağlık Örgütü tarafından antimikrobiyel direnç mekanizmaları nedeniyle yeni nesil antibiyotiklerin hayvanlarda, bitkilerde ve akuakültürde kullanılmaması, mevcut antibiyotiklerin kullanımının sınırlandırılması önerilmektedir. Antibiyotiklerin amacı dışında kullanımı; gıda zincirinde antibiyotik rezidü sorunu oluşturmakta, duyarlı bireylerde sağlık problemlerini artırmakta ve kamunun kıt kaynaklarının tüketilmesine neden olmaktadır. Pasif immunoterapi yoluyla antimikrobiyel kalıntı ve direnç gelişim mekanizmalarının bertarafı sayesinde özellikle riskli dönemlerde antibiyoterapiye alternatif olarak kullanılabilecek bir mücadele yöntemidir. Araştırmamızda Salmonella-free olarak temin edilen Ross 308 damızlık tavuklar kullanıldı. Salmonella gallinarum, Salmonella pullorum, Salmonella gallinarum + Salmonella pullorum şeklinde Freund’un Adjuvanı kullanılarak aşı hazırlandı ve 21 gün arayla 2 aşı uygulaması yapıldı. Aşılanan tavuklardan yumurtalar günlük olarak toplandı ve +4°C’de muhafaza edildi. Amonyum sülfat çöktürme ve diyaliz ile IgY saflaştırıldı. Toplam protein miktarı Bradford yöntemi ile tayin edildi. Elde edilen proteinler SDS-PAGE ile tanımlandı. Daha sonra yapılan in vitro etkinlik denemelerinde tüm fraksiyonların mikrobiyel büyümeyi inhibe ettiği görüldü. Gruplardan özellikle Salmonella gallinarum + Salmonella pullorum antikorlarının etkinliğinin yüksek olduğu gözlendi. Araştırmamız sonucunda, yumurta sarısı antikorlarının kısa süre içerisinde elde edilebileceği, Salmonella dışında da etken spesifik antikor üretilebileceği ve mikrobiyel büyümeyi inhibe edici yönleriyle antibiyotiklere alternatif olarak kullanılabileceği bir yöntem ve süreç geliştirildi. Anahtar Kelimeler: Antikor üretimi, Bakteriyel Büyüme İnhibisyonu, Immunoglobulin Y, Immunoterapi, Salmonella. 2. SUMMARY Isolation of Immunoglobulin Y from Hen Egg and Determination of Its Efficiency The World Health Organization recommends not to use the new age antibiotics on animals, plants and aquaculture because of their antimicrobial resistance mechanisms. It further recommends to restrict the use of existing antibiotics. The misuse of antibiotics causes a problem of antibiotic residual in the food chain. It also causes an increase in health problems of individuals, a decrease of effectiveness and reduces low public budgetary. Passive immunotherapy that counteracts pathogens helps destroying antimicrobial residual and resistance mechanisms. It can be used even in the presence of infection so it is a good alternative for antibiotic therapy, especially in hazardous periods. In our research a Salmonella-free Ross 308 parent stock was used. Immunization was achieved using Salmonella gallinarum, Salmonella pullorum, Salmonella gallinarum+Salmonella pullorum and Freund's Adjuvant. Two doses of immunization was given for 21 days. Eggs were collected daily and saved. The amount of total protein was measured by Bradford method. Ammonium sulfate precipitation and dialysis methods were used for IgY purification.Proteins were identified by SDS-PAGE. Subsequently in vitro activity of all fractions in inhibiting microbial growth was determined. Especially Salmonella gallinarum + Salmonella pullorum antibodies were observed to have high activity. In conclusion, we have developed a method and process whereby egg yolk antibodies can be obtained in a short time and can be produced as specific antibodies against other antigens than Salmonella. Key Words: Antibody Generation, Bacterial Growth Inhibition, Immunoglobulin Y, Immunotherapy, Salmonella