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Altered luteal expression patterns of genomic and non-genomic progesterone receptors in bitches at different reproductive states
(Elsevier Inc., 2024) Uçar, Eyyüp Hakan; Peker, Cevdet; Hitit, Mustafa; Köse, Mehmet; Tatar, Musa; Bozkaya, Faruk; Atlı, Mehmet Osman
The binding of steroid hormones to their specific receptors is necessary to exert their effects on target cells. Progesterone (P4), a steroid hormone, carries out its effects through both genomic and non-genomic (the cell membrane-associated) receptors. This study aimed to ascertain luteal expression patterns of genomic and non-genomic progesterone receptors in bitches in physiological (early dioestrus and early pregnant) and pathological (pyometra) reproductive states. Luteal tissue was collected from the bitches at early dioestrus (ED, n = 5), early pregnant (EP, n = 5), and pyometra (PY, n = 5). The expression profiles of Steroidogenic Acute Regulator Protein (STAR), Progesterone Receptor (PGR), Membrane Progestin Receptors (PAQR5, PAQR7 and PAQR8), and Progesterone Membrane Components (PGMRC1 and PGMRC2) were examined at the mRNA levels using Real-Time Polymerase Chain Reaction (RT-PCR). Protein levels of PGR, PGMRC1 and PGMRC2 were detected by western blotting (WB). The STAR expression was found in all groups, with a statistical difference observed between EP and PY groups (P < 0.05). The protein level of PGR was determined to be highest in the EP group and lowest in the PY group. The expression of PAQR8 increased in the EP group (P < 0.05). The PAQR5 exhibited high expression in the EP group and low expression in the PY group (P < 0.05). PGRMC1 was more elevated in the EP group and lower in the PY group (P < 0.05). Protein levels of PGMRC1 and PGMRC2 were also observed at the highest expression in EP group. According to the altered expression profiles for examined receptors, we suggest that those progesterone receptors have roles in early pregnancy or pyometra in bitches.
Bazı kaba yemlere ilave edilen probiyotiklerin in vitro organik madde sindirimi ve metan üretimi üzerine etkileri
(2019) Kaplan, Oktay; Bozkaya, Faruk; Güler, Ali Murat
Bu çalışma ruminantlarda yaygın olarak kullanılan bazı kaba yemlere katılan probiyotiklerin (Lactobacillus rhamnosus, Bifidobacterium lactis ve Saccharomyces boulardii) in vitro ortamda metan gazı oluşumuna etkisini tespit etmek için yapılmıştır. Bu amaçla %0.1 oranında probiyotik ilave edilen öğütülmüş kaba yem örnekleri rumen sıvısı içeren özel cam tüpler içerisinde 39 °C’de 24 saat inkube edilmiştir. İnkubasyon sonunda oluşan toplam gaz içerisindeki metan (CH4) gazı ve karbondioksit (CO2) yüzdesi CH4 ölçüm cihazı yardımıyla belirlenmiştir. Ayrıca her bir deneme grubundaki in vitro organik madde sindirilebilirliği (IVOMS), amonyak azotu miktarı (NH3-N), metabolik enerji (ME) ve pH değerleri belirlenmiştir. Buğday samanına ilave edilen B. lactis’in oluşan toplam gaz, CH4 ve CO2 hacmini ve IVOMS’ni düşürdüğü, S. boulardii’nin ise CH4 yüzdesini yükseltirken, CO2 yüzdesini düşürdüğü gözlenmiştir. Çayır kuru otuna ilave edilen L. rhamnosus oluşan toplam gaz miktarını, CH4 miktarını ve IVOMS’ni yükseltirken CH4 yüzdesini etkilememiştir. Silaj ve yonca kuru otuna ilave edilen probiyotikler CH4 ve CO2 düzeylerini etkilememiştir. Sonuç olarak çalışmada buğday samanına katılan B. lactis dışındaki probiyotik mikroorganizmalar CH4 üretimini arttırmış ya da etkilememiştir. Buğday samanına ilave edilen B. lactis’in CH4 miktarını azaltması, Çayır kuru otuna ilave edilen L. rhamnosus’un ise CH4 miktarını arttırmasının söz konusu yemlerin IVOMS’ni etkilemesinden kaynaklandığı, bu nedenle sunulan çalışmada kullanılan probiyotiklerin CH4 miktarını azaltmakta etkili olmadığı sonucuna varılmıştır.
Cell-specific expression pattern of toll-like receptors and their roles in animal reproduction
(Springer Science and Business Media Deutschland GmbH, 2022) Atlı, Mehmet Osman; Hitit, Mustafa; Özbek, Mehmet; Köse, Mehmet; Bozkaya, Faruk; 0000-0001-9853-5334; 0000-0001-5636-061X; 0000-0003-0070-8458
Toll-like receptors (TLRs), a part of the innate immune system, have critical roles in protection against infections and involve in basic pathology and physiology. Secreted molecules from the body or pathogens could be a ligand for induction of the TLR system. There are many immune and non-immune types of cells that express at a least single TLR on their surface or cytoplasm. Those cells may be a player in a defense system or in the physiological regulation mechanisms. Reproductive tract and organs contain different types of cells that have essential functions such as hormone production, providing an environment for embryo/fetus, germ cell production, etc. Although lower parts of reproductive organs are in a relationship with outsider contaminants (bacteria, viruses, etc.), upper parts should be sterile to provide a healthy pregnancy and germ cell production. In those areas, TLRs bear controller or regulator roles. In this chapter, we will provide current information about physiological functions of TLR in the cells of the reproductive organs and tract, and especially about their roles in follicle selection, maturation, follicular atresia, ovulation, corpus luteum (CL) formation and regression, establishment and maintenance of pregnancy, sperm production, maturation, capacitation as well as the relationship between TLR polymorphism and reproduction in domestic animals. We will also discuss pathogen-associated molecular patterns (PAMPs)-induced TLRs that involve in reproductive inflammation/pathology.
Early pregnancy diagnosis using a commercial ELISA test based on pregnancy-associated glycoproteins in Holstein-Friesian heifers and lactating cows
(Tubitak Scientific & Technological Research Council Turkey, 2016) Kaya, Mehmet Salih; Kose, Mehmet; Bozkaya, Faruk; Mutlu, Hasan; Ucar, Eyyup Hakan; Atli, Mehmet Osman
In this study, we aimed to investigate the efficacy of a commercial ELISA test kit for detecting pregnancy-associated glycoproteins (PAGs) in peripheral blood for early pregnancy diagnosis and compare plasma PAG levels during early pregnancy in both Holstein-Friesian heifers and lactating cows. A total of 231 plasma samples were collected from heifers and lactating cows on days 25, 28, and 32 after insemination. Pregnancies were confirmed 30 days after the collection of plasma samples. Plasma PAG levels were measured using a commercial ELISA test kit for diagnosing bovine pregnancy. The effects of examination date and animal status (heifers vs. lactating cows) on PAG levels in heifers and lactating cows were analyzed using two-way ANOVA. Plasma PAG levels were significantly higher in heifers than in lactating cows; levels also increased significantly with the date of examination, i.e. days 25, 28, and 32 for heifers (P < 0.001), but not for lactating cows (P > 0.05). Although the sensitivity, specificity, and accuracy of pregnancy diagnosis using the ELISA test were acceptable in both groups, the performance of the test was superior in pregnant heifers compared to lactating cows.
Effect of long term heat stress and dietary restriction on the expression of small heat shock protein (sHSP) genes in rat liver tissue
(2016) Güzeloğlu, Aydın; Aydilek, Nurettin; Bozkaya, Faruk; Kayış, Seyit Ali; Atlı, Mehmet Osman; Kaya, Mehmet Salih
Amaç: Bu çalışmanın amacı uzun süreli sıcaklık stresi ve yem kısıtla-masının rat karaciğer dokusunda bazı küçük ısı şoku protein (sHSP) genlerinin mRNA düzeyindeki ekspresyonu düzeyleri üzerine etkisi-nin araştırılmasıdır. Gereç ve Yöntem: Bu amaçla on haftalık yaştaki toplam 24 Spra-gue-Dawley rat 4 gruba ayrıldı. Grup I ve Grup II'deki ratlar 22°C'lik ortam sıcaklığında, Grup III ve Grup IV'teki ratlar ise 38°C'lik ortam sıcaklığında tutuldu. Grup I ve III'teki ratlar ad libitum olarak beslen-di, Grup II ve IV'teki ratlara ise ad libitum grupların tükettiği yemin %60'ı kadar yem verildi. Uygulama 9 hafta sürdürüldükten sonra karaciğer doku örnekleri alınarak sıvı azot içerisinde donduruldu ve RNA izolasyonuna kadar muhafaza edildi. Doku örneklerinden total RNA izole edildikten sonra HspB1, HspB5, HspB6, Hsp10 ve Hsp11 genlerinin ekspresyon düzeyleri gerçek zamanlı nicel polimeraz zin-cir reaksiyonu (RT-qPCR) yöntemi ile incelendi.Bulgular: Sıcaklık stresi HspB2, HspB8 ve Hsp70 genlerinin ekspres-yonunu önemli ölçüde arttığı, HspB1, HspB5, HspB6, Hsp10 ve Hsp11 genlerinin ekspresyonunu ise etkilemediği belirlendi. Yem kısıtla-ması HspB6 geninin expresyonunu arttırırken HspB1, HspB2, HspB5, HspB8 HspB10, HspB11 ve Hsp70 genlerinin ekspresyonunu etkile-mediği gözlendi. Uygulamalar arasında interaksiyon gözlenmedi.Öneri: Çalışmanın sonuçları uzun süreli sıcaklık stresinin rat kara-ciğer dokusundaki sHSP genlerinin ekspresyonlarını değişik düzey-lerde etkilediğini, yem kısıtlamasının sHSP genlerinin sıcaklık stresi tarafından etkilenen ekspresyonlarını değştirmediğini göstermiştir.
Effects of long-term heat stress and dietary restriction on the expression of genes of steroidogenic pathway and small heat-shock proteins in rat testicular tissue
(Blackwell Publishing Ltd, 2017) Bozkaya, Faruk; Atlı, Mehmet Osman; Güzeloǧlu, Aydın; Kayış, Seyit Ali; Yıldırım, Mehmet Erol; Kurar, Ercan; Yılmaz, Rahşan; Aydilek, Nurettin
The aim was to investigate the effects of long-term heat stress and dietary restriction on the expression of certain genes involving in steroidogenic pathway and small heat-shock proteins (sHSPs) in rat testis. Sprague Dawley rats (n = 24) were equally divided into four groups. Group I and II were kept at an ambient temperature of 22°C, while Groups III and IV were reared at 38°C for 9 weeks. Feed was freely available for Group I and Group III, while Group II and Group IV were fed 60% of the diet consumed by their ad libitum counterparts. At the end of 9 weeks, testicles were collected under euthanasia. Total RNA was isolated from testis tissue samples. Expression profiles of the genes encoding androgen-binding protein, follicle-stimulating hormone receptor, androgen receptor, luteinising hormone receptor, steroidogenic acute regulatory protein (StAR), cyclooxygenase-2 and sHSP genes were assessed at mRNA levels using qPCR. Long-term heat stress decreased the expression of StAR and HspB10 genes while dietary restriction upregulated StAR gene expression. The results suggested that long-term heat stress negatively affected the expression of StAR and HspB10 genes and the dietary restriction was able to reverse negative effect of heat stress on the expression of StAR gene in rat testis.
Expression patterns of Toll-like receptors in the ovine corpus luteum during the early pregnancy and prostaglandin F2?-induced luteolysis
(Elsevier Science Inc, 2018) Atli, Mehmet O.; Kose, Mehmet; Hitit, Mustafa; Kaya, M. Salih; Bozkaya, Faruk
The aim of this study was to elucidate the expression profiles of Toll-like receptors (TLRs) in the ovine corpus luteum (CL) during early pregnancy and prostaglandin F2 alpha (PGF2 alpha)-induced luteolysis. For this purpose, multiparous Anatolian Merino ewes were selected and randomly allotted into cyclic (including those in the induced luteolysis group, n = 20) and pregnant (n = 12) groups. All of the ewes were scheduled to be slaughtered for predetermined days/hour during the estrous cycle, early pregnancy, and PGF2 alpha induced luteolysis. The CLs were collected from both cyclic and pregnant ewes on days 12 (C12 and P12; n = 8) and 16 (C16 and P16; n = 8) and pregnant ewes on day 22 (P22; n = 4). For the induced luteolysis model, ewes were injected with PGF2 alpha on day 12 of the estrous cycle and CLs were collected at 1 h (PG1h; n = 4), 4 h (PG4h; n = 4), and 16 h (PG16h, n = 4) after injection. Quantitative polymerase chain reaction (VCR) was used to evaluate the expression profiles of TLR2, TLR4, TLR6, TLR8, and TLR10, while free-floating in situ hybridization and immunohistochemistry were used to define the spatial localization of TLR2, TLR4, and TLR7 in the CL. Data were then analyzed by one-way ANOVA and were considered statistically significant when P values were lower than 0.05. Expression of TLR2 was upregulated in both early and late stages of luteolysis (P < .05). An upregulation of TLR4 was detected at PG16h, while TLR6 was decreased at PG4h (P < .05). Expression of TLR7 and TLR8 was significantly increased during early pregnancy, at both PG16h and regressed groups (C16, P < .05). In contrast, 711210 was downregulated during PGF2 alpha-induced luteolysis and on P16 (P < .05). TLR4 and TLR7 proteins were particularly localized in endothelial cells on C12/PGOh, but prominent signals corresponding to TLR4 and TLR7 were detected in luteal cells at PG16h. The results suggest an involvement of TLRs in the luteolytic mechanism in ovine CL, as indicated by differential expression levels of TLRs during PGF2 alpha-induced luteolysis. Moreover, the present study indicates that early pregnancy-mediated changes in TLR expression in the CL may contribute to the establishment and maintenance of ovine pregnancy. (C) 2018 Elsevier Inc. All rights reserved.
Expression profiles of interferon-tau stimulated genes (ISGs) in peripheral blood leucocytes (PBLs) and milk cells in pregnant dairy cows
(Kafkas Üniversitesi Veteriner Fakültesi, 2014) Köse, Mehmet; Görgülü, Mehmet; Kaya, Mehmet Salih; Aydilek, Nurettin; Bozkaya, Faruk; Bayrıl, Tahir; Kurar, Ercan; Kıyma, Zekeriya; Güzeloğlu, Aydın; Atlı, Mehmet Osman
In previous reports, it was indicated that measurement of activity of Interferon-tau Stimulated Genes (ISGs) in Peripheral Blood Leucocytes (PBLs) may be used as an alternative early pregnancy detection method in dairy cows. However, there are no data showing the expression profiles of ISGs in other body fluids containing leucocytes such as milk. In the present study, it was hypothesized that leucocytes in milk samples may reflect the increases in expression profiles of ISGs as shown in PBLs. For this purpose, nine pregnant lactating Holstein cows were used. Insemination day was accepted as day zero (day 0). Blood and milk samples were collected on day 0 and 18 after insemination for cell isolation. Total RNA was extracted from isolated cells and converted to cDNA. Steady state levels of Interferon-tau Stimulated Gene 15 (ISG15), Myxovirus (influenza virus) resistance 1 (MX1) and 2 (MX2) mRNA transcripts were assayed by using real- time reverse transcriptase PCR. Relative Expression Software Tool (REST2009) was used for statistical analyses. There was no statistical significant difference for expression levels of ISG15, MX1 and MX2 mRNAs between days 0 and 18 in milk samples. However, when compared to day 0, levels of ISG15 and MX2 transcripts were increased 6.97±0.68 fold and 5.84±1.27 fold on day 18 in PBLs in pregnant cows, respectively (P<0.05). According to this result, it may be suggested that milk cells are not suitable measurement of expression profiles of ISGs to detect early pregnancy in lactating dairy cows.
IMPACTS OF SPECIFIC CRYOPROTECTANTS ON SPERM FREEZING AND RELATIONSHIPS BETWEEN CRYODAMAGE AND OXIDATION STRESS PARAMETERS IN AWASSI RAM SPERM
(Cryo Letters, 2021) Varisli, Omer; Erat, Serkan; Bozkaya, Faruk; Aydilek, Nurettin; Taskin, Abdullah
BACKGROUND: The role of oxidative stress during cryoprotectant treatment has received little attention. OBJECTIVE: To assess the effects of different cryoprotectants and discover relationships between cryodamage and oxidative stress parameters on Awassi ram sperm. MATERIALS AND METHODS: The sperm samples diluted with Salamon's tris-citrate (TRIS) containing 20% centrifuged egg yolk and 0.5, 1.0 or 1.5 M Glycerol (Gly), methanol (M), 2-methoxyethanol (2-ME), dimethylacetamide (DMA) and 1.2 propanediol (PR). After 2 h of equilibration at +4 degrees C, the sperm samples were frozen in liquid nitrogen vapour and stored. RESULTS: The best post-thaw motility (43.3%, 41.7%) of sperm was achieved when protected with 0.5 and 1.0 M glycerol. Arylesterase and ceruloplasmin parameters were significantly different after equilibration, whereas sulfhydryl groups were significantly different after freezing in their respective groups (P< 0.05). CONCLUSION: The increased use of glycerol caused greater loss of motility. The role of oxidative stress in freezing was also found to be limited.
Impacts of specific cryoprotectants on sperm freezing and relationships between cryodamage and oxidation stress parameters in awassi ram sperm
(Cryo Letters, 2021) Varışlı, Ömer; Erat, Serkan; Bozkaya, Faruk; Aydilek, Nurettin; Taşkın, Abdullah
BACKGROUND: The role of oxidative stress during cryoprotectant treatment has received little attention. OBJECTIVE: To assess the effects of different cryoprotectants and discover relationships between cryodamage and oxidative stress parameters on Awassi ram sperm. MATERIALS AND METHODS: The sperm samples diluted with Salamon's tris-citrate (TRIS) containing 20% centrifuged egg yolk and 0.5, 1.0 or 1.5 M Glycerol (Gly), methanol (M), 2-methoxyethanol (2-ME), dimethylacetamide (DMA) and 1.2 propanediol (PR). After 2 h of equilibration at +4 degrees C, the sperm samples were frozen in liquid nitrogen vapour and stored. RESULTS: The best post-thaw motility (43.3%, 41.7%) of sperm was achieved when protected with 0.5 and 1.0 M glycerol. Arylesterase and ceruloplasmin parameters were significantly different after equilibration, whereas sulfhydryl groups were significantly different after freezing in their respective groups (P< 0.05). CONCLUSION: The increased use of glycerol caused greater loss of motility. The role of oxidative stress in freezing was also found to be limited
Influence of humic acid addition to drinking water on laying performance and egg quality in Japanese quails
(Agricultural Research Communication Centre, 2018) Kaplan, Oktay; Avci, Mehmet; Denek, Nihat; Baran, Murat Sedat; Nursoy, Huseyin; Bozkaya, Faruk
The experiment was performed to investigate the effect of humic acid to drinking water on the laying performance and quality of eggs in Japanese quails. Hundred and forty, 9-weeks old female Japanese quails (Coturnix coturnix japanica) were randomly designated to four treatment groups forming 5 replicates each containing seven birds. Group I was the control group. Group II, III, IV were administered 250 ml/ton, 350 ml/ton, 450 ml/ton humic acid into drinking water, respectively. The egg characteristics were determined after 8th and 10th weeks of experiment. Addition of humic acid had a positive effect on the live weight change (p < 0.05) and yolk weight (p < 0.01). Egg shell thickness, albumen weight and yolk color were significantly (p < 0.01) lower in the experimental groups than that of control group. However humic acid had no effect on egg production, feed consumption, feed efficiency and other egg quality. In conclusion, data of our study demonstrated that liquid humate applied in drinking water has significantly improved live weight change and yolk weight. The positive effects of liquid humates, possibly related to improved nutrient utilization through various metabolic activities in the body are yet to be further investigated.
Kilis, Halep ve Kıl Keçilerinde Beta-Kazein (CSN2) Genindeki Çeşitliliğin Allel Spesifik PCR, Real-Time PCR ve Sekans Analizi ile Araştırılması
(2019) Atlı, Mehmet Osman; Yiğin, Akın; Bozkaya, Faruk
Bu çalışmada Kilis, Halep ve Kıl keçilerinde Beta-kazein geninin (CSN2) promotor bölgesi ile 7. eksonunun belirli nükleotitlerindeki farklılıkların allel spesifik polimeraz zincir reaksiyonu (AS-PCR), Real-Time PCR ve sekans analizi yöntemleriyle araştırılarak söz konusu keçi populasyonlarının bu açıdan genetik çeşitliliğinin belirlenmesi amaçlanmıştır. Bu amaçla toplam 246 DNA örneği kullanılmıştır. CSN2 geninin promotor bölgesindeki g1311T>C mutasyonu AS-PCR ile, 7. eksonun 404. pozisyonundaki C>T mutasyonunun (CSN2C alleli) varlığı ise Real-Time PCR ve sekans analizi ile incelenmiştir. AS-PCR işlemi sonucunda CSN2 geni promotor bölgesi 1311. nükleotitte T allelinin sıklığı bütün populasyonlarda yüksek bulunmakla birlikte Kıl keçilerinde C allelinin sıklığı (0.0489) Halep (0.0190) ve Kilis (0.0188) keçilerinden daha yüksek bulunmuştur. Bütün populasyonlar değerlendirildiğinde 7. eksonunun 404. pozisyonundaki T nükleotitinin (CSN2C alleli) sıklığının (0.824) C nükleotitinden (0.176) daha yüksek olduğu gözlenmiştir. En yüksek CSN2C alleli sıklığı Kıl keçilerinde (0.8864) en düşük ise Kilis keçilerinde (0.6364) gözlenmiştir. Sonuç olarak Kilis, Halep ve Kıl keçilerinde CSN2 geninin promotor bölgesindeki g1311T>C mutasyonu ile 7. eksonun 404. pozisyonundaki C>T mutasyonu açısından çeşitlilik gösterdiği tespit edilmiştir. Bu çeşitliliğin keçi sütünün bileşimi ve beta kazein içeriğini etkileyip etkilemediğinin belirlenmesi için daha fazla araştırmaya ihtiyaç bulunmaktadır.
Koyunlarda Maternal Plazmadan Fötal Cinsiyet Tayininde Fenol-Kloroform İzolasyon Metodu İle İki Farklı Ticari İzolasyon Kitinin Kullanılabilirliği
(2016) Dursun, Şükrü; Köse, Mehmet; Bozkaya, Faruk; Bayrıl, Tahir; Atlı, Mehmet Osman; Kaya, Mehmet Salih; Kırbaş, Mesut
Koyunlarda fötal cinsiyetin belirlenmesinde kullanılan yöntemlerden biri maternal plazmadaki fötal DNA'ların Polimeraz Zincir Reaksiyonu (PZR) ile çoğaltılmasına dayanır. Ancak, maternal plazmadan izole edilen DNA miktarı ve kalitesi, kullanılan DNA izolasyon yöntemine göre önemli ölçüde değişebilmektedir. Bu çalışmada, üç farklı DNA izolasyon metoduyla koyun maternal plazmasından izole edilen DNA'ların PZR ile çoğaltılarak fötüs cinsiyetlerinin belirlenmesi amaçlandı. Bu amaçla, gebeliğin 20. haftasındaki 13 adet koyundan plazma örnekleri alındı ve doğum sonrası yavruların cinsiyetleri doğrulandı. Pozitif kontrol amacıyla iki adet koç plazması, negatif kontrol amacıyla ise iki adet gebe olmayan koyun plazması kullanıldı. Maternal plazmadan fenol-kloroform izolasyon metodu ve iki farklı ticari nükleik asit izolasyon kiti (QIAamp ve FitAmp) kullanılarak DNA izole edildi. İzole edilen DNA örneklerinden X ve Y kromozomları üzerindeki Amelogenin ve SRY (Sex-determining Region Y) genlerinin hedef bölgeleri PZR yöntemi ile çoğaltıldı. Ticari nükleik asit izolasyon kitleriyle elde edilen DNA'lardan SRY ve Amelogenin primerleri ile yapılan farklı PZR işlemlerinde fötüs cinsiyetinin belirlenmesine yönelik spesifik PZR ürünleri elde edilemedi. Diğer taraftan, fenol-kloroform izolasyon metoduyla izole edilen DNA örneklerinden 10'unda (7 erkek ve 3 dişi fötal DNA içeren örnekte) fötüs cinsiyeti doğru bir şekilde teşhis edildi (10/13). Ancak erkek fötüs taşıyan üç koyundan elde edilen DNA örneklerinden erkek cinsiyet teşhisine özgü PZR ürünleri elde edilemedi (3/13). Çalışmanın sonuçları koyunlarda maternal plazmadan DNA izolasyonunda iki farklı ticari nükleik asit izolasyon kiti kullanıldığında fötal cinsiyetin tespit edilemediğini, diğer taraftan fenol-kloroform izolasyon metodu ile fötal cinsiyet teşhisi yapılabildiğini, ancak bu metodun da hatalı sonuçlara neden olabildiğini göstermiştir
Plasma profile of Pregnancy Associated Glycoproteins during Postpartum period in Konya Merino ewes lambing single and twin
(Univ Zulia, Facultad Ciencias Veterinarias, 2023) Ucar, Eyyup Hakan; Kose, Mehmet; Kirbas, Mesut; Bozkaya, Faruk; Atli, Mehmet Osman
This study investigated the plasma profile of Pregnancy Associated Glycoproteins (PAGs) after lambing in Konya Merino ewes using specific cattle pregnancy test kit. A total of 16 Konya Merino ewes were used as a material. Four groups were set up, ewes birthing a male lamb (SM group, n=4), a female lamb (SF group, n=4), twin male lambs (TM group, n=4) or twin female lambs (TF group, n=4). Blood plasma samples were taken on days 0 (lambing day), 3, 7, 10, 14 and then weekly until day 35. All samples were analysed using a bovine pregnancy test kit to detect of PAGs level. It was determined that there was a strong negative correlation between PAGs levels in peripheral blood and days after lambing (r2=0.969; P<0.01). However, no relationship was found between PAGs level and lamb gender or birth type. In conclusion, plasma PAGs level decreases rapidly in Konya Merino ewes regardless of lamb gender and birth type. Moreover, the results showed that the plasma profile of PAGs after lambing in Konya Merino ewes can be monitored with a bovine commercial ELISA-based pregnancy test kit, and the test results can be used in decisions and assessments based on the levels of PAG molecules.
Rating of putative housekeeping genes for quantitative gene expression analysis in cyclic and early pregnant equine endometrium
(Elsevier Science Bv, 2011) Kayis, Seyit A.; Atli, Mehmet O.; Kurar, Ercan; Bozkaya, Faruk; Semacan, Ahmet; Aslan, Selim; Guzeloglu, Aydin
The aim was an evaluation of a set of housekeeping genes (HKGs) to be used in the normalization of gene expression in the equine endometrium. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), hypoxanthine ribosyl transferase 1 (HPRT1), ubiquitin B (UBB), tubulin alpha 1 (TUBA1), ribosomal protein L32 (RPL32), beta-2-microglobulin (B2M), 18S rRNA (18S), and 285 rRNA (28S) HKGs were evaluated using real-time PCR and were compared in different physiological stages of the endometrium. Endometrial biopsies were obtained from mares on day of ovulation (d0, n = 4), at late diestrus (LD, n = 4), after luteolyis (AL, n = 4) of the cycle and on days 14 (P14; n = 3), 18 (P18, n = 3) and 22 (P22; n = 3) of pregnancy. A model based on REML with support of descriptive statistics was proposed in accordance with experimental design and was further confirmed with principal component analysis (PCA). Results were compared with widely used software including geNorm, BestKeeper, and NormFinder. Results indicated that GAPDH was the most stable HKG and RPL32 was ranked as the second best. 18S and 285 were found to be the least stable. The proposed model, PCA, geNorm, and BestKeeper were in agreement in detecting the most stable and the least stable HKGs in the equine endometrium during the estrous cycle and early pregnancy. (C) 2011 Elsevier B.V. All rights reserved.
RELATIONSHIP BETWEEN TOXICITY OF CRYOPROTECTANTS, OSMOTIC AND OXIDATIVE STRESSES IN AWASSI RAM SPERM
(Cryo Letters, 2022) Varisli, Omer; Bozkaya, Faruk; Aydilek, Nurettin; Taskin, Abdullah
BACKGROUND: The relationship between the toxicity of cryoprotectants and their osmotic and/or oxidative stresses remains to be further investigated. OBJECTIVE: To investigate the toxic effects of different cryoprotectants and osmotic stress on Awassi ram sperm and to determine the relationship between oxidative and antioxidative status of the sperm. MATERIALS AND METHODS: Pooled sperm samples were exposed to sucrose solutions of different concentrations (75 to 900 mOsm) and isosmotic condition (290-325 mOsm) was re-established by adding HEPES buffered Tyrode's lactate. Sperm samples were mixed with 0.5, 1.0 and 1.5 M of glycerol, methanol, 2-methoxyethanol, dimethylacetamide or 1,2-propanediol for 5 min and returned to isosmotic condition. Sperm samples were exposed to cryoprotectants at 4 degrees C for 2 hours and isosmotic conditions were re-established. Motility, viability, acrosome integrity and oxidative or antioxidative parameters were determined. RESULTS: Treatment with hypo- or hyperosmotic sucrose solution reduced motility and viability without affecting acrosome integrity. The addition and removal of glycerol and dimethylacetamide (1.0 or 1.5 M) decreased sperm motility, while cryoprotectants had no effect on viability except for 1.5 M glycerol. Chilling significantly reduced the motility and viability of the sperm, but not the acrosome integrity. Rapid addition or removal of cryoprotectants also did not affect the acrosome integrity. Cryoprotectants changed only the ceruloplasmin level, while there were significant post-chilling differences in lipid hydroperoxide, paraoxonase and ceruloplasmin levels. CONCLUSION: Cryoprotectants without other additives have limited protection and glycerol can be toxic to spermatozoa. The oxidative stress plays a role in cryoprotectant toxicity and chilling stress.

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