Determination of novel reference genes for improving gene expression data normalization in selected canine reproductive tissues – a multistudy analysis

Background Real time RT-PCR (qPCR) is a useful and powerful tool for quantitative measurement of gene expression. The proper choice of internal standards such as reference genes is crucial for correct data evaluation. In female dogs, as in other species, the reproductive tract is continuously undergoing hormonal and cycle stage-dependent morphological changes, which are associated with altered gene expression. However, there have been few attempts published so far targeted to the dog aimed at determining optimal reference genes for the reproductive organs. Most of these approaches relied on genes previously described in other species. Large-scale transcriptome-based experiments are promising tools for defining potential candidate reference genes, but were never considered in this context in canine research. Results Here, using available microarray and RNA-seq datasets derived from reproductive organs (corpus luteum, placenta, healthy and diseased uteri) of dogs, we have performed multistudy analysis to identify the most stably expressed genes for expression studies, in each tissue separately and collectively for different tissues. The stability of newly identified reference genes (EIF4H, KDELR2, KDM4A and PTK2) has been determined and ranked relative to previously used reference genes, i.e., GAPDH, β-actin and cyclophillin A/PPIA, using RefFinder and NormFinder algorithms. Finally, expression of selected target genes (luteal IL-1b and MHCII, placental COX2 and VEGFA, and uterine IGF2 and LHR) was re-evaluated and normalized. All proposed candidate reference genes were more stable, ranked higher and introduced less variation than previously used genes. Conclusions Based on our analyses, we recommend applying KDM4A and PTK2 for normalization of gene expression in the canine CL and placenta. The inclusion of a third reference gene, EIF4H, is suggested for healthy uteri. With this, the interpretation of qPCR data will be more reliable, allowing better understanding of canine reproductive physiology

Učinak nepravilne primjene intravaginalnog umetka za otpuštanje progesterona na plodnost holštajnsko-frizijske krave: prikaz slučaja.

In this case report, the surgical removal of a progesterone-releasing intravaginal device (PRID®, 1.55 g of progesterone) found adhered to the uterus was performed, and its effect on fertility in a Holstein-Friesian cow evaluated. No data older than six months was available on the cow; however, during the preceding sixmonth period, the animal showed no signs of estrus. Interestingly, in the course of a reproductive examination, a PRID-like spiral mass was identified on the right uterine horn. Following diagnosis of the mass, surgical removal was considered. Right flank laparotomy was selected as the surgical entry site. Approximately one month after surgery, the cow showed signs of estrus, but no insemination was performed until the third cycle. After insemination, the cow become pregnant and completed the gestation period. Therefore, we concluded that the PRID may have caused anestrus, and that right flank laparotomy was a safe procedure for the removal of a PRID from the uterus, and particularly to recover fertility, in this case.U prikazu slučaja opisano je kirurško uklanjanje intravaginalnog umetka za otpuštanje progesterona (PRID®, 1,55 g progesterona). Umetak je bio pričvršćen za maternicu krave holštajnsko-frizijske pasmine u koje je obavljena kontrola plodnosti. U prethodnom šestomjesečnom razdoblju nije bilo podataka o kravi, a tijekom sljedećih šest mjeseci kontrole nije bilo znakova estrusa. Tijekom pregleda reprodukcijskih organa, u desnom rogu maternice otkrivena je spiralna tvorevina, slična umetku. Nakon postavljene dijagnoze razmatrana je mogućnost kirurškog uklanjanja. Odabrana je laparotomija s pristupom na desnoj bočnoj strani. Približno mjesec dana nakon operacije kod krave su opaženi znakovi estrusa, no osjemenjivanje nije obavljeno prije trećeg ciklusa. Nakon osjemenjivanja, krava je postala bređa s normalnim trajanjem gravidnosti. Autori zaključuju da PRID može biti uzrokom izostanka estrusa kod krava. Laparoskopska operacija s desne bočne strane je sigurna metoda za uklanjanje PRID-a iz maternice, posebno s obzirom na uspostavljenu plodnost krave u ovom prikazu

A spatially uniform illumination source for widefield multi-spectral optical microscopy

Illumination uniformity is a critical parameter for excitation and data extraction quality in widefield biological imaging applications. However, typical imaging systems suffer from spatial and spectral non-uniformity due to non-ideal optical elements, thus require complex solutions for illumination corrections. We present Effective Uniform Color-Light Integration Device (EUCLID), a simple and cost-effective illumination source for uniformity corrections. EUCLID employs a diffuse-reflective, adjustable hollow cavity that allows for uniform mixing of light from discrete light sources and modifies the source field distribution to compensate for spatial non-uniformity introduced by optical components in the imaging system. In this study, we characterize the light coupling efficiency of the proposed design and compare the uniformity performance with the conventional method. EUCLID demonstrates a remarkable illumination improvement for multi-spectral imaging in both Nelsonian and Koehler alignment with a maximum spatial deviation of ~1% across a wide field-of-view

Promjene fizikalnih i kemijskih pokazatelja u mlijeku koza pasmine damask tijekom ranog postpartalnog razdoblja

The aim of this study was to reveal the milk components and the changes in their characteristics in Damascus goats during the early postpartum period. For this purpose, 50 Damascus goats (aged 2 to 8 years) were selected for this study. A total of 291 milk samples were collected under sterile conditions during the mornings of the 10th, 20th and 30th days postpartum. Results were statistically analysed using the Mann-Whitney U Test and repeated measures ANOVA. All collected samples were analysed for milk composition (lactose (L), protein (P), casein (C), fat (F), total solid (TS), solid not fat (SNF), freezing point (FP), acidity (A), density (D), free fatty acid (FFA), and citric acid (CA)). A significant decrease was detected in P, C, F, TS and SNF values between the 10th and 30th days postpartum (pp) (P <0.01). Similar results were also observed in FP, A, D and CA values between the same days (P <0.05; P <0.01). A significant positive correlation was detected between FP, A, P, C, F and TS (0.812 and 0.980; P<0.05 and P<0.01). Additionally, a correlation was detected between C and P (0.983; P <0.01). These results reveal that these parameters decreased significantly in the early puerperal period compared to the first 10 days, and these decreases were not associated with an increase during milk yield in the early period of lactation (up to the 30th-day pp).Cilj rada bio je prikazati sastojke mlijeka i njihove promjene tijekom ranog postpartalnog razdoblja u koza pasmine damask. Istraživanjem je obuhvaćeno 50 koza u dobi od 2 do 8 godina. Ukupno 291 uzorak mlijeka prikupljen je pod sterilnim uvjetima, tijekom jutra 10., 20. i 30. dana postpartalnog razdoblja. Rezultati su statistički analizirani Mann-Whitney U testom i ANOVA metodom s ponovljenim mjerenjima. Svi prikupljeni uzorci mlijeka analizirani su s obzirom na sastojke odnosno pokazatelje: laktoza (L), protein (P), kazein (C), mast (F), ukupna suha tvar (TS), bezmasna suha tvar (SNF), točka smrzavanja (FP), kiselost (A), gustoća (D), slobodne masne kiseline (FFA) i limunska kiselina (CA). Znakovito sniženje (P<0,01) između 10. i 30. postpartalnog razdoblja uočeno je u vrijednostima P, C, F, TS i SNF. Slični rezultati (P<0,05; P<0,01), između istih dana, utvrđeni su i za FP, A, D te CA. Statistički znakovita pozitivna korelacija (P<0,05 i P<0,01; 0,812 i 0,980;) uočena između FP, A, P, C, F i TS. Osim toga, potvrđena je korelacija između C i P (0,983; P<0,01). Rezultati pokazuju da su se razine istraženih sastojaka/pokazatelja tijekom ranog puerperija znakovito snižavale u odnosu na razdoblje prvih 10 dana. Navedeno snižavanje nije povezano s povećanjem koje se javlja tijekom proizvodnje mlijeka u ranom razdoblju laktacije (do 30. dana postpartalnog razdoblja)

Evaluation of disease and medication knowledge levels of diabetic patients

Background and Aims: Diabetes mellitus is a public health problem with a significant economic burden to society. Patient attitude is a significant determinant of adherence and clinical outcomes. We aimed to evaluate the disease and medication knowledge level of diabetic patients and possible patient-related predictive factors. Methods: Adult diabetic patients were included in the study. Patients' demographic and health-related data were collected using a pre-prepared form. Two questionnaires, KAP knowledge tool and Medication Assessment tool were used to assess the disease and medication knowledge levels of patients. Results: The mean age of 159 patients was 54.44 & PLUSMN;12.24 years. The disease and medication knowledge scores were high with an average of 13.9 & PLUSMN;1.74 and 5.26 & PLUSMN;0.53, respectively. Higher scores were linked to being male (p=0.042; p=0.007) and higher educational status (p<0.001; p=0.006). Lower scores were recorded in patients with comorbidities (p=0.002; p<0.001), older patients (p<0.001), longer disease duration (p<0.001; p=0.009), longer antidiabetic drug use (p<0.001; p=0.009) and using more drugs (p=0.002; p=0.006). None of the patients could mention any possible side effects of their medications. Conclusion: It can be deduced that patient-related factors are significant predictors of patients' disease and medication knowledge. Patient-specific education in addition to rational pharmacological intervention is necessary to achieve better clinical outcomes

Relevance of components of Investment Model and some variables in close relationships with high and low self-monitoringYakın ilişkilerde Yatırım Modeli unsurlarının ve bazı değişkenlerin yüksek ve düşük izlenim ayarlamacılığı ile ilgisi

The aim of this study is to investigate the relationship between some variables related to close relationships (expectation of the relationship duration, loyalty to the relation, finding the partner attractive, evaluating the quality of the relationship positively, having issues in the relationship, reliance on the relationship, the wish that she/he had never started the relationship, finding the relationship important, happiness with the partner) and the components of Investment Model (relationship satisfaction, quality of alternatives, relationship investment) with self-monitoring which is a variable related to the self concept. For this aim, married or not, 472 participants which have a romantic relationship, completed a personal information form, Snyder’s Self-monitoring Scale (Bacanlı, 1990; Snyder, 1974) and Relationship Stability Scale (Büyükşahin, Hasta,&amp; Hovardaoğlu, 2005; Rusbult, Martz, &amp;Agnew,1998). A logistic regression is conducted to investigate whether the components of Investment Model are relevant to any self-monitoring groups (high self-monitoring or low self-monitoring). Results indicated that quality of alternatives (positively) and having issues in the relationship (negatively) are related to high self-monitoring. Results are discussed in terms of theoretical approaches. ÖzetBu araştırmada yakın ilişkilerle ilgili bazı değişkenlerin (ilişki süresine ilişkin beklenti, ilişkiye bağlılık, partneri çekici bulmak, ilişkinin niteliğini olumlu değerlendirmek, ilişkide sorun yaşamak, ilişkide güvende hissetmek, ilişkiye hiç başlamamış olmayı istemek, ilişkiyi önemli bulmak, partnerle mutluluk) ve Yatırım Modeli’nin (Investment Model) unsurları (ilişki doyumu, seçeneklerin niteliğinin değerlendirme, ilişki yatırımı) ile benlik kavramıyla ilişkili izlenim ayarlamacılığı (self-monitoring) değişkeninin ilgili olup olmadığının test edilmesi amaçlanmıştır. Bu amaçla, (evli ya da evli olmayan) bir romantik ilişkisi bulunan 472 kişiye kişisel bilgi formu, Kendini Ayarlama Ölçeği (Bacanlı, 1990; Snyder, 1974) ve İlişki İstikrarı Ölçeği (Büyükşahin, Hasta ve Hovardaoğlu, 2005; Rusbult, Martz ve Agnew,1998) uygulanmıştır. Yapılan lojistik regresyon analizi ile araştırma kapsamında ele alınan ve yakın ilişkilerle ilgili değişkenlerin ve Yatırım Modeli unsurlarının yüksek ya da düşük izlenim ayarlamacılığı gruplarından herhangi biriyle ilgili olup olmadığı test edilmiştir. Analiz sonucunda seçeneklerin niteliğini değerlendirme (pozitif olarak) ve ilişkide sorun yaşama  (negatif olarak) değişkenlerinin yüksek izlenim ayarlamacılığı ile ilgili olduğu bulunmuştur. Sonuçlar kuramsal yaklaşımlar çerçevesinde tartışılmıştır

The effects of Meloxicam application on quality of life and development in calves

Although it is stated that there is an increase in calf loss rates at the global level and especially in modern dairy farms, this issue, which is an international animal welfare issue, has not been adequately defined at the national, large farm, or small farm level in most developing Countries. The rate of calf deaths before, during, and after birth is between 2–20% in heifers and cows. Previous studies showed that 75% of deaths occur during birth or within the first hour after birth, 10% before birth, and 15% within 48 hours after birth. This study aimed to determine the effect of Meloxicam administration on the quality of life and development of calves. An experimental method was used to achieve the aim of this research. A total of 60 Holstein calves, 30 in the experimental group and 30 in the control group, were included in the study for the study group. MELOXICAM (0.5 mg·kg-1) was administered to the mother cow and the calf immediately after birth, 2 days before the expected delivery. In this study, an 8 week process was passed to collect the data. It was concluded that Meloxicam did not affect the BHBA values of maternal cows. No calves died from pneumonia and diarrhea–related diseases after being treated with Meloxicam. From the total protein and total albumin values of the calves, it was concluded that Meloxicam did not affect the protein ratio, but did affect the albumin ratio. It was concluded that Meloxicam application increased the vital activity of calves

Organik Üzüm Yetiştiriciliği

Bu projenin amacı Güneydoğu Anadolu Bölgesinde üzüm yetiştiriciliğinde bitki besleme ile hastalık ve zararlılara karşı organik tarımın uygun gördüğü preparatları kullanarak organik üzüm yetiştiriciliğine veri tabanı oluşturmaktır. Araştırma, Gaziantep/Oğuzeli ilçesinde bulunan, çiftçi bağında 2002-2009 yılları arasında, yüksek goble şekli verilen ve 4 x 4 m dikilmiş Dımışkı üzüm çeşidi bağında yürütülmüştür. Çalışmada çiftlik gübresi, yeşil gübre ve konvansiyonel uygulamalar karşılaştırılmıştır. Her parselden ayrı ayrı toprak alınmış ve analiz yapılmıştır. Konvansiyonel üretimin yapıldığı parsele üreticilerin kullandığı ve pratikte kullanılan kimyasal gübreler uygulanmıştır. Bağda külleme hastalığı ve salkım güvesine karşı mücadele yapılmıştır. Uygulama alanlarında dekara verim, dane ağırlığı, SÇKM ve serbest asitlik değerlerine bakılmıştır. Verim yönünden yıllar itibariyle yapılan analiz sonucunda istatistiksel olarak fark çıkmamakla birlikte, yeşil gübre uygulanan alandan ortalama 481,63 kg/da verim alınırken, çiftlik gübresi uygulanan alandan 474,75 kg/da, Konvansiyonel alandan ise 454,10 kg/da verim alınmıştır. Sonuç olarak yeşil gübre uygulamasının maliyetinin düşük, organik olarak yetiştirilen üzüm fiyatının yüksek olması, yeşil gübre uygulamasını diğer uygulamalara göre ön plana çıkarmıştır. Organik üzüm yetiştiriciliği konusunda farklı besleme ve mücadele materyali ile farklı yöntemler alternatif olarak araştırma konusu yapılarak çalışmalara devam edilmelidir. Özellikle organik ürünün talep ve fiyat durumunu belirleyecek araştırma çalışmaları yapılmalıdır

The Effects of Separate and Combined Use of PGF2α and GnRH Hormones and the Addition of Βeta-Carotene on Fertility Parameters in Dairy Cows with Ovarian Cysts

Background: Ovarian cysts are commonly observed pathologies, which interfere with normal cyclic activity and adversely affect fertility in cows. Beta-carotene is effective in the reduction of reproductive problems by inducing the natural defence mechanisms of the body. There are several methods that can be used for the treatment of ovarian cysts. The separate and combined use of GnRH and PGF2α commonly uses in the treatment of ovarian cysts. Therefore, in the presented study the effects of Beta-carotene (βC) addition for the treatment of ovarian cysts either with GnRH solely or GnRH and PGF2α in combination on the fertility parameters of dairy cows were investigated.Materials, Methods &amp; Results: Seventy-six Holstein Friesian cows having ovarian cysts diagnosed by ultrasonography (USG) were divided into three groups. Cows in Group I (GI, n = 27), were injected with GnRH (Buserelin acetate, 5 mL, im), PGF2α (Tiaprost-trometamol, 5 mL, im) and βC (20 mL/cow, into 4 regions by im route). In Group II (GII, n = 25) GnRH (Buserelin acetate, 5 mL, im) and PGF2α (Tiaprost-trometamol, 5mL, im) were administrated while GnRH (Buserelin acetate, 5 mL, im) solely in Group III (GIII, n = 24). Cysts were monitored via USG, and blood samples were collected on the on day of treatment (day 0) and on the 7th and 14th days following the administrations. Cows shoving oestrous were inseminated and pregnancy diagnoses were performed on the 40th day following insemination. Treatment results showed that there were statistically no significant differences between GI and GII (P &gt; 0.05). Only numerical difference obtained in time from therapy to pregnancy and overall pregnancy index (P &gt; 0.05). Overall pregnancy rate (85 %), first service pregnancy rates (40 %) and overall pregnancy index (2.11) in GI were found significantly higher than GIII (53.3 %; 20 %; 4.12) [P &lt; 0.05]. No significant difference was observed in progesterone (P4) levels between the groups (P &gt; 0.05). It was found that βC administrations significantly increased βC levels in GI than GII and GIII on the 7th and 14th days (P &lt; 0.05).Discussion: One of the most common problems encountered in modern dairy production is the development of ovarian cysts. Treatments for ovarian cyst are numerous and variable, and have changed considerably over the years. In the present study, GnRH and PGF2α were administered together as a combination, and as a result of this combined use, higher percentages were obtained for both pregnancy rate and fertility parameters in GI and GII, in comparison to the group administered with GnRH alone (GIII). Better outcome from combination therapy (GnRH and PGF2α) may be due to the fact that luteal thickening in the walls of cysts was determined by ultrasonography, but P4 values were not identified immediately and the treatments were not categorized in accord with these values. No statistically significant differences were determined between GI and GII concerning the fertility parameters investigated, however numerical and proportional differences were observed. βC levels were significantly higher on day 7 and 14 after treatment in GI which were administered βC additionally to the treatment protocol for ovarian cysts. This statistical difference suggests that administration of βC in combination therapy is also effective in the treatment of ovarian cysts. In conclusion, it was determined that high pregnancy rates were obtained by the combined treatment of ovarian cysts (GnRH + PGF2α) and number of inseminations per conception were at desired limits. Better percentile and numerical fertility parameters were achieved in the group, which additionally received βC, high numbers of infertility cases, βC supplementation could be a viable option for treatment