Seminal plasma (SP) contains several types of compounds derived from the epididymides and accessory glands. The aim of this study was to examine the protein composition of different ejaculate fractions. Trial I: fractionated ejaculates were collected from two normal and two subfertile stallions. Samples containing pre-sperm fluid and the first sperm-rich jets (HIGH-1), the main sperm-rich portion (HIGH-2), the jets with low sperm concentrations (LOW), and a combined whole-ejaculate (WE) sample was centrifuged, and the SP was filtered and frozen. A part of each SP sample was stored (5°C, 24 h) with spermatozoa from HIGH-2 and skim milk extender. Sperm motility was evaluated after storage in extender mixed with the stallion's own SP or SP from one of the other stallions (sperm from a normal stallion stored in SP from a subfertile stallion and vice versa). Protein composition was analysed using reverse-phase liquid chromatography (RP-HPLC), N-terminal sequencing and mass spectrometry. The area-under-the-curve (AUC) was used for quantitative comparison of proteins within fractions. Trial II: semen samples were collected from seven stallions. Fractions with the highest (HIGH) and lowest (LOW) sperm concentrations and WE samples were examined using SDS-PAGE and densitometry. No significant differences emerged between fractions in the AUC-values of the Horse Seminal Protein-1 (HSP-1) and HSP-2 peaks, or the peak containing HSP-3 and HSP-4 (HSP-3/4). Levels of HSP-1, HSP-2 and HSP-3/4 were not significantly correlated with total sperm motility, progressive sperm motility or average path velocity after storage. Significant differences between ejaculate fractions in the amount of different protein groups present in SP were not found in Trial I; but in Trial II, the proteins in the 60-70 kDa range were more abundant in LOW than in HIGH and WE, indicating that this band contained proteins derived mainly from the seminal vesicles, which produce most of the SP in LOW.
Seminal plasma (SP) is a mixture of contents from the testes, epididymides and accessory sex glands. The sperm concentration is highest in the first few jets, or fractions, of the ejaculate, and the composition of SP varies between these fractions because accessory gland secretions are released in a specific order. The aim of this study was to compare the levels of Na, Cl, K, Mg, Ca, inorganic phosphate (Pi) and the enzymes alkaline phosphatase (AP), acid phosphatase (ACP) and β-glucuronidase (BG) in the different fractions of the ejaculate and in different stallions. All semen collections were done using a computer-controlled phantom that collects the ejaculatory jets separately in five cups. The cups with the highest (HIGH) and the lowest (LOW) sperm concentration were analysed. In Trial I, semen was collected from three reproductively normal stallions. In Trial II, ejaculates of two reproductively normal stallions were compared to those of two subfertile stallions. In Trial III, semen was collected from seven stallions with varying reproductive history. The sperm-rich fractions contained the highest levels of AP, ACP, BG and inorganic phosphate, and the values were positively correlated to the sperm concentration. Significant differences between the subfertile and the fertile stallions pairs in HIGH:LOW ratios were found in Pi and Cl concentrations. The highest concentrations of Ca and Mg were found in the last fractions with low sperm concentrations, with no significant differences between the fertile and the subfertile stallion pairs. The concentrations of K, Na and Cl were similar in HIGH and LOW fractions and in whole ejaculate samples. Pre-sperm fluid contained the highest concentrations of Na and Cl. Some of the possible variation in storage tolerance between ejaculates and ejaculatory fractions could perhaps be explained by differences in the composition of SP.
-Pregnancy rates after frozen semen inseminations (AI), particularly in older and problem mares, are lower than after fresh semen AI. Uterine contractility and the inflammatory reaction after frozen semen insemination were studied in two groups of mares: the abnormal group comprised of 6 old barren mares categorized in biopsy category IIB or III, and the control group including 6 reproductively normal young maiden mares in biopsy category I or IIA. All 12 mares were inseminated in the first cycle with 2 mL of phosphate-buffered saline (PBS) and in their second cycle with 2 mL of frozen semen containing 800 × 10 6 spermatozoa. Before and 1, 2, 4, 8, and 20 to 24 h after this treatment, all mares were examined by ultrasonography for intrauterine fluid accumulations (IUFA). The examinations were videotaped to count the number of uterine contractions later. Uterine fluid was obtained by tampon before treatment, and by the tampon method followed by uterine lavage after the last examination. Fluids were cultured bacteriologically, and polymorphonuclear leukocytes (PMN) were counted. Trypsin-inhibitor capacity (TIC), lysozyme concentration, and β-glucuronidase (BGase) and N-acetyl-β-D-glucosaminidase (NAGase) activities were determined in frozen-thawed tampon and lavage fluids. Both treatments induced significant neutrophilia in the uterine lumen. Although PMN concentrations were numerically higher after frozen semen AI than after PBStreatment, the difference was not significant. There was not any difference between the mare groups either. The amount of IUFA differed only in the normal group between frozen semen AI and PBS treatment, and between 0-and 24-h samples for frozen semen AI. Although abnormal mares showed consistently more fluid than normal mares, this difference was not significant. Uterine contractions and enzyme concentrations between groups did not differ. None of the variables showed significant differences between the normal and abnormal mares in their reaction to frozen semen AI.horse / insemination / frozen semen / uterine fluid / inflammation / leukocyte
Bu çalýşmada üreme mevsimi dýşýnda bulunan Tuj ýrký koyunlarda Testosteron antikoru, β-karoten ve E vitamini uygulamalarý ile çoğul gebeliğin artýrýlmasý ve MDA (Malondialdehit) düzeylerinin azaltýlmasý amaçlanmýştýr. Testosteron antikoru kastre edilmiş Tuj ýrký bir koçtan elde edildi. Çalýşma 29 adet koyunda 3 grup halinde yapýldý. Grup I'deki koyunlara (n=10) vaginal süngerlerin uygulanmasýndan 1 hafta önce testosteron antikoru, β-karoten ve E vitamini uygulandý. Grup II'deki (n=9) koyunlara testosteron antikoru uygulandý. Grup III'deki (n=10) koyunlar ise kontrol grubunu oluşturdu. Koyunlardaki östrus oranlarý; I. Gruptaki koyunlarda %90.0, II. Gruptaki koyunlarda %88.9 ve III. Gruptaki koyunlarda %100.0 olarak belirlendi. Gebelik oranlarý ise Grup I, II ve III'de sýrasýyla %40.0, %33.3 ve %50.0 oranýnda belirlendi. Grup I ve Grup II'deki koyunlarda çoğul gebelik elde edilirken kontrol grubu koyunlarda çoğul gebelik elde edilmedi. Gebe ve gebe olmayan tüm koyunlarda vaginal sünger uygulamasýndan 28 gün sonra plazma MDA düzeylerinin arttýğý belirlendi ancak gruplar arasýnda istatistiksel olarak önemli bir fark tespit edilmedi (P>0.05). Sonuç olarak testosteron antikoru, β-karoten ve E vitamini uygulamasýnýn çoğul gebelik görülme oranýnda artýşa neden olduğu görüldü.
Detection of the circulating tumour cells (CTC) in dogs with a mammary tumour is a useful tool to reveal the micrometastases long before metastases are recognised clinically. The aim of this study was to evaluate the association of the epidermal growth factor receptor (EGFR), claudin 7 (CLND7) and epithelial cell adhesion molecule (EPCAM) with the clinical indices and to reveal the diagnostic importance of these biomarkers in canine mammary tumours (CMTs). Peripheral blood (PB) samples were collected from 45 bitches (group MT) which had single mass with malignant epithelial tumours and 9 healthy bitches (group H). Real time PCR (rt-PCR) was performed to determine the expression levels of EGFR, CLDN7, and EPCAM. Mean values of EGFR and CLDN7 expressions were significantly higher in group MT compared to group H (P < 0.01 and P < 0.001, respectively). The expression level of CLDN7 was positively correlated with EGFR and EPCAM (P < 0.001 and P < 0.05, respectively). The EPCAM expression was associated with increased tumour size (P < 0.05) and EPCAM tended to decrease in the presence of skin ulceration on tumour (P = 0.05). Furthermore, expression levels of EGFR in intact dogs were significantly higher compared to spayed dogs in group MT (P < 0.01). The EGFR expression was significantly higher in the presence of metastases (P < 0.05). Also, increased EGFR was determined in grade 2 compared to grade 1 (P < 0.05). In conclusion, these results show that EGFR, CLDN7, EPCAM markers are measureable in PB and they may provide valuable information about the clinical pathophysiology of CMT.
Ö ÖZ ZE ET T A Am ma aç ç: : Bu çalışmada; sağlıklı ve meme tümörlü köpeklerde beyaz kan hücreleri [white blood cell (WBC)], platelet (PLT) ve kandan izole edilen total ribonükleik asit (RNA) miktarlarının karşılaştırılması ve ayrıca; tümör tipi, kitle büyüklüğü, kitle sayısı, yaş, ülserasyon ve metastaz varlığı gibi klinik bulgular ile belirtilen hematolojik parametreler arasındaki ilişkinin ortaya konulması amaçlandı. G Ge er re eç ç v ve e Y Yö ön nt te em ml le er r: : Araştırmanın materyalini, ortalama 5,94±1,21 yaşında 9 adet sağlıklı dişi köpek ve ortalama 11,19±0,41 yaşında 45 adet meme tümörlü dişi köpek oluşturdu. Bunun yanında, kısırlaştırma amacıyla kliniğimize getirilen köpekler ise kontrol grubu olarak kullanıldı. Operasyon öncesi dönemde hemogram ve total RNA izolasyonu amacıyla kan örnekleri alındı. Total RNA izolasyonu kit prosedürüne uygun olarak gerçekleştirildi. B Bu ul lg gu ul la ar r: : Çalışma ve kontrol grubundaki köpekler yaş, PLT, WBC, total RNA miktarı açısından değerlendirildi. Bunun sonucunda; yaş (p<0,001), PLT (p<0,05), total RNA (p<0,05) miktarları meme tümörlü köpeklerde daha yüksek miktarda ölçüldü. Ayrıca, meme tümörlü köpeklerde WBC değerleri ile kitle büyüklüğü (p<0,05) ve total RNA miktarları (p<0,001) arasında pozitif korelasyon saptandı. Bunun yanı sıra, ülsere tümörlü köpeklerde WBC miktarının daha yüksek olduğu belirlendi (p<0,01). S So on nu uç ç: : Bu çalışmada; PLT miktarının karsinojenik olgularda arttığı tespit edildi. Ayrıca, meme tümörlü köpeklerde prognostik tümör belirteçlerinin (biyomarkerların) saptanabilmesi için kandan izole edilen total RNA miktarının, WBC miktarı ile ilişkili olduğu sonucuna varıldı. Elde edilen bulguların tümör patofizyolojisi ile ilgili moleküler düzeydeki çalışmalara katkı sağlayacağı düşünülmektedir.A An na ah h t ta ar r K Ke e l li i m me e l le er r: : Kan trombositleri; lökositler; meme neoplazmaları; total RNA A AB BS S T TR RA AC CT T O Ob bj je ec ct ti iv ve e: : The amount of white blood cell (WBC), platelet (PLT) and total ribonucleic acid (RNA) isolated from blood in healthy bitches and bitches with mammary tumors (MT) were compared in this study. Also, determination of the relationship between hematological parameters and clinical findings such as tumor type, mass size, number of mass, age, ulceration and metastasis was aimed. M Ma at te er ri ia al l a an nd d M Me et th ho od ds s: : The material of the research was composed of the data belonging to the 9 healthy bitches approximately 5.95±1.21 years old and 45 bitches with MT approxiamtely 11.19±0.41 years old. Control group included the bithces which were presented to our clinic for ovariohysterectomy. Blood samples were collected in preoperative term for total RNA isolation and hemogram. Total RNA isolation was performed according to kit procedure. R Re es su ul lt ts s: : Bitches in study and control group were evaluated in terms of age, PLT, WBC and total RNA values. Consequently, age (p<0.001), PLT (p<0.05) and total RNA (p<0.05) values were detected higher in bitch...
Özet: Çalışmanın amacı, kısırlaştırılmış kedilerde krom pikolinatın (CrPic) morfometrik parametreler, ağırlık artışı ile serum lipit ve leptin seviyeleri üzerine etkisinin belirlenmesidir. Kısırlaştırılıp, 2 ay adaptasyon süresini takiben çalışmaya dahil edilen 32 (22 dişi ve 10 erkek) adet, yaşları 1-5 arasında değişen ve ağırlık ortalaması 3021 ± 453 g olan kediler 4 gruba ayrıldı. Kontrol grubu olarak tutulan kediler (Kontrol I), günlük enerji gereksinimi (60 kcal ME/kg) hesaplanarak ticari kuru mama ile beslendi. Ad-libitum beslenen diğer gruptaki kedilere sırasıyla CrPic kapsülden 0 (Kontrol II), 200 ya da 600 µg Cr/gün, 16 hafta süreyle uygulandı. En fazla ağırlık artışının belirlendiği Kontrol II'de, deri kalınlığı, abdominal çevre ve leptin düzeyinde düşük oranda bir artış gözlenirken, vücut kitle indeksi (BMI) ve göğüs çevresindeki artış önemli bulundu (P<0.05). CrPic verilen gruplarda ise bu değişkenlerdeki azalma önemli değildi. Plazma leptin seviyesi ile vücut ağırlığı (r = 0.510, P<0.01) ve aterojenik indeks (AI) arasında pozitif korelasyon (r = 0.479, P<0.05) belirlenirken, Cr arasında negatif korelasyon (r =-0.445, P<0.05) belirlendi. Kontrol II'de trigliserit (TG), total kolesterol (TC), düşük dansiteli lipoprotein (LDL), LDL ve yüksek dansiteli lipoprotein (HDL) oranı ve AI değerleri en yüksek seviyede olmakla birlikte, gruplar arasında fark gözlenmedi. Krom ilavesinin de bu parametrelere önemli bir etkisinin olmadığı saptandı. Serum Cr ve total kolesterol düzeyleri arasında negatif ilişki belirlendi (r =-0.468, P<0.05). Sonuç olarak, kısırlaştırıldıktan sonra ad-libitum beslenen tüm gruplarda canlı ağırlık, BMI, deri altı yağ doku ve plazma leptin seviyesinde artış belirlenirken, krom ilavesinin bu parametrelere önemli bir etkisinin olmadığı belirlendi.
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