CRISPR-Of-Things%253A Applications and Challenges of the Most Popular Gene Editing Tool in the Fields of Health, Agriculture and Environment

Almost all cells of any living organism contain DNA, a hereditary molecule that passes from generation to generation during reproduction. The term quot%253Bgenomequot%253B generally refers to the total DNA sequences in an organism. The genome consists of DNA sequences called gene, which plays a role in the basic biological processes involved in many phenotypic and genotypic characteristics, such as performing cellular functions, controlling numbers and species, regulating energy production, metabolism, and combating diseases. Gene editing is the process of pre-designing and modifying a particular DNA sequence in a targeted gene. The most widely used technique is CRISPR-Cas technology. For this purpose, the DNA helix is ​​cut at a certain point, to form a double-strand break (DSB), and naturally existing cellular repair mechanisms repair the DSB. Modes of the repair mechanisms may affect the gene function. When DSB is formed, gene editing techniques can be applied to remove, insert, or replace a newly modified sequence using a synthetic donor template DNA. In developed and developing countries, CRISPR-Cas studies in addition to research and development studies are rapidly increasing. In addition to increasing population, changing weather conditions, declining farmland, increasing biotic and abiotic stresses are other important barriers to agricultural production, food, and feed supply. In this report, CRISPR-Cas applications are introduced in detail from the studies that carried out gene modifications in the fields of health, animals, plants, microorganisms, and food supply. Besides, these technologies and applications have been examined in terms of world biosafety legislation and the scientific risk assessment of the products developed using the CRISPR-Cas technique

Çiğ köftede Listeria monocytogenes'in gelişimi

Çiğ köfte (raw meat ball), a traditional food of Turkey, has become a commercial success in recent decades. This study was undertaken to determine the growth of Listeria monocytogenes in çiğ köfte at 4 ºC and 25 ºC. For this purpose çiğ köfte samples were divided into four groups and contaminated with 103 cfu/g, 104 cfu/g and 105 cfu/g, respectively. Also there was a control group for detecting the contamination of L. monocytogenes. Samples were analyzed at 0, 4, 8, 12 and 24 hours by using Most Probable Number (MPN) Technique. The results were shown that there was a 1-2 log MPN/g increase within 8 hours in samples which were incubated at 25 ºC and also after 24 hours of incubation 2 log MPN/g increase were observed in all groups. In Group “A” L. monocytogenes level increased to 5.66 log MPN/g from the initial number of 2.97 log MPN/g within 24 hour. Similarly L. monocytogenes levels increased to 6.17 log MPN/g from 4.32 log MPN/g and 7.04 log MPN/g from the initial number of 5.17 log MPN/g in Group “B” and Group “C” within 8 hour period at 25 ºC, respectively. The results of this study indicate that if the contamination occur during the production process of çiğ köfte with L. monocytogenes, çiğ köfte should be considered as an important source of public health problems, even its initial contamination level is low. As the lack of legislation on çiğ köfte, it is necessary to establish microbiological standards by regarding the measures of European Community Standards also hygienic measures must be taken to ensure a wholesome product.Geleneksel bir ürün olan çiğ köfte son yıllarda daha çok ticari önem kazanarak geniş bir tüketim alanına sahip olmuştur. Bu çalışmada 4 ºC ve 25 ºC’lerde muhafaza edilen çiğ köftelerde L. monocytogenes gelişiminin belirlenmesi amaçlanmıştır. Çalışmada çiğ köfte örnekleri biri kontrol olmak üzere 4 gruba ayrılmıştır. Birinci grup 103 kob/g, ikinci grup 104 kob/g ve üçüncü grup 105 kob/g düzeyinde L. monocytogenes (NCTC-2167) ile kontamine edildikten sonra 0., 4., 8., 12. ve 24. saatlerde USDA/FSIS tarafından önerilen yöntem ile çiğ köftelerdeki L. monocytogenes gelişimi analiz edilmiştir. L. monocytogenes’in düzeyinin belirlenmesinde ise Most Probable Number (MPN) tekniği kullanılmıştır. Çalışmada 25 ºC’deki üç grupta da ilk 8 saat içerisinde 1-2 log MPN/g düzeyinde artış olduğu, 24 saat sonunda ise bu artışın 2 log MPN/g düzeyine ulaştığı saptanmıştır. Bu kapsamda A grubunda 2.97 log MPN/g olan başlangıç kontaminasyon düzeyi 25 ºC’de 24 saat sonunda 5.66 log MPN/g seviyesine yükselmiştir. B ve C gruplarında ise sırasıyla 4.32 log MPN/g ve 5.17 log MPN/g’dan 8. saat sonunda 6.17 log MPN/g ve 7.04 log MPN/g düzeyine ulaşmıştır. Çalışma bulgularından elde edilen sonuçlar ışığında yetersiz hijyenik koşullar altında üretilen çiğ köftelerde düşük düzeyde dahi L. monocytogenes ile bir kontaminasyon durumunda halk sağlığı riski oluşabileceği bu nedenle üretimden tüketime kadar her aşamada asgari hijyenik şartlar ile uygun muhafaza koşullarının sağlanması gerektiği düşünülmektedir. Bu amaca yönelik olarak, Türkiye’de çiğ köftelerin de yer aldığı hazır gıdalara yönelik yasal düzenlemelerin Avrupa Birliği mevzuatı dikkate alınarak hazırlanmasının halk sağlığı ve gıda güvenliği açısından yararlar sağlayabileceği görüşüne varılmıştır

The effect of ascorbic acid, storage period and packaging material on the formation of volatile N-nitrosamine in sausages

This study aims to determine the amounts of seven volatile N-nitrosamine (VNA) derivatives which are in the risk group, in processed sausages. It also aimed to investigate the effects of the amount of added ascorbic acid, on the VNA level during the sausage manufacturing processes. For this purpose, meat doughs were prepared with two different levels of ascorbic acid. These meat doughs were put into the production process and packaged in two different packages. Thus, VNA derivatives and their amounts were determined according to production stages (heat treatments), packaging method (vacuum, MAP), storage process (Day 1, 7, 15, 30, 60, 90). As a result, it was found that the sausage product carries risk of VNA formation from the beginning of its production until the last day of storage before consumption for up to 90 days

Yumurta tavuklarında gentamisinin karşılaştırmalı farmakokinetiği

Çalışmada, yumurtlayan tavuklara damar içi, kas içi ve deri altı yolla verilen gentamisin sülfatın (5 mg/kg canlı ağırlık) karşılaştırmalı farmakokinetiğinin belirlenmesi amaçlandı. 24 tavuğa ilaç uygulandıktan sonra kan örnekleri 0 (uygulama öncesi), 0.083, 0.166, 0.25, 0.5, 0.75, 1, 2, 4, 8, 12, 24, 36 ve 48. saatlerde toplandı. Gentamisin konsantrasyonları Avrupa Birliği tarafından önerilen yöntemde modifikasyon yapılarak belirlendi. Gentamisinin toplam konsantrasyonu (C1, C2 ve C1a) ölçüldü. En düşük belirlenme limiti 0.01 ?g/ml olarak hesaplandı. Non-kompartmental farmakokinetik analizleri Excel ile çalışan program olan PK solver ile belirlendi. Damar içi uygulamayı takiben plazma konsantrasyonu zaman eğrisinin altında kalan alan (AUC0-?), ilk hız atılma sabitesi (?z), yarı-ömür (t1/2?z) ve ortalama tutulma zamanı (MRT) sırasıyla 224.46 ?g/mL saat, 0.06 saat-1, 11.52 saat ve 9.50 saat olarak ölçüldü. Kas içi ve deri altı uygulamadan sonra ortalama maksimum plazma konsantrasyonu (Cmax) sırasıyla26.64 ve 36.92 ?g/mL, maksimum konsantrasyona ulaşmak için geçen zaman (Tmax) ise aynı (0.75 saat) olarak belirlendi. Yine kas içi ve deri altı uygulamadan sonra sırasıyla t1/2?z 8.35 ve 8.24 saat, MRT ise 11.05 ve 9.79 saat olarak ölçüldü. Kas içi ve deri altı uygulama arasında Cmax değerleri hariç ve damar içi ile diğer uygulamalar arasında t1/2?z değerleri hariç tutulursa önemli farklılıklar olmadığı sonucuna varılmıştır.The aim of this study was to compare pharmacokinetics of gentamicin sulphate (5 mg/kg body weight) after single intravenous, intramuscular and subcutaneous administration in laying hens. Blood samples were collected at time 0 (pretreatment), and at 0.083, 0.166, 0.25, 0.5, 0.75, 1, 2, 4, 8, 12, 24, 36 and 48 h after drug administration in 24 laying hens. Gentamicin concentrations were determined using the HPLC method recommended by the European Union by some modifications. The total concentration of the gentamicin (C1, C2 and C1a) was calculated. The lowest detection limit was 0.01 μg/ml. Noncompartmental pharmacokinetic analyses were performed using Excel add-in program, PK solver. Following IV administration the area under the plasma time-concentration curve from time zero to infinity (AUC0-∞), first-order elimination rate constant (λz), terminal half-life (t1/2λz) and mean residence time (MRT) were 224.46 μg/mL h, 0.06 h-1, 11.52 h and 9.50 h, respectively. After i.m. and s.c. dosing, the mean maximum plasma concentrations (Cmax) were 26.64 and 36.92 μg/mL, achieved at a same post-injection times (Tmax) of 0.75 h, respectively. The t1/2λz was 8.35 and 8.24 h, the MRT was 11.05 and 9.79 h, respectively, after IM and SC administration. There are no significant between IM and SC administration excluding the Cmax values and between i.v. and other administration excluding the t1/2λz values

In vitro Effects of Chitosan on the Survival of Listeria monocytogenes

The nonlinear robust stability theory of Georgiou and Smith (IEEE Trans. Auto. Control, 42(9):1200--1229, 1997) is generalized to the case of notions of stability with bias terms. An example from adaptive control illustrates non trivial robust stability certificates for systems which the previous unbiased theory could not establish a non-zero robust stability margin. This treatment also shows that the BIBO robust stability results for adaptive controllers in French (IEEE Trans. Auto. Control, 53(2):461--478, 2008) can be refined to show preservation of biased forms of stability under gap perturbations. In the nonlinear setting, it also is shown that, in contrast to LTI systems, the problem of minimizing nominal performance is not equivalent to maximizing the robust stability margin

In Vitro Effects of Phthalate Mixtures on Colorectal Adenocarcinoma Cell Lines

KUZUKIRAN, OZGUR/0000-0001-9294-2801; Filazi, Ayhan/0000-0002-2800-6215WOS: 000368384400003PubMed: 26081030Among endocrine-disrupting chemicals, phthalates are an important concern because of their widespread exposure in humans and environmental contamination. Even though the use of some phthalates has been restricted for toys, some plastics, and food contact materials, exposure to the mixture of these contaminants at very low concentrations in various matrices are still being reported. In the current research, the effects of the mixture of some phthalates were studied. Di-n-butyl phthalate (DBP), n-butyl benzyl phthalate (BBP), di-2-ethylhexyl phthalate (DEHP), diisononyl phthalate (DiNP), di-n-octyl phthalate (DNOP), and diisodecyl phthalate (DiDP) were tested on two colorectal adenocarcinoma cell lines; DLD-1 and HT29 were studied as described before. Cells were treated with increasing log concentrations (0.33 ppt to 33.33 ppb) of the phthalate mixture, cell viability/proliferation was measured by MTT and staining with neutral red and crystal violet; lactate dehydrogenase (LDH) activity was measured following 24-h exposure. Cell viability/proliferation increased from phthalate treatment at concentrations less than 33.33 ppt. The phthalate mixture induced increases in HT29 proliferation of 10.94% at 33.33 ppt and 60.87% at 3.33 ppt, whereas this proliferation relation at lower concentrations was not found for DLD1 cells. The present study demonstrates preliminary information regarding the low dose induction of proliferation of the cancer cells by phthalate mixtures. Because non-monotonic dose responses are still being debated, further studies are required to re-evaluate the reference doses defined by governments for phthalates