Determination of expression patterns of miR-26a, and preimplantation factor levels for early pregnancy detection in nulliparous and multiparous cows.

For maximum productivity in a dairy farm, the earliest and the most accurate detection of pregnancy is essential. The aim of this study was to determine the efficacy of expression patterns of miR-26a, and serum Preimplantation Factor (PIF) levels for pregnancy diagnosis during the early pregnancy in nulliparous and multiparous cows. A total of 60 cows (30 nulliparous and 30 multiparous Holstein cows) were enrolled in the study. Blood samples were collected for miR-26a on days 8 and 16 (D8 and D16), and for the PIF on days 10 and 20 (D10 and D20) following insemination (D0). Pregnancies were determined by ultrasonography on the 28th day after insemination. Expression levels of miR-26a determined by qPCR. PIF levels were assessed by using commercial ELISA kits. All data were analyzed by using the MIXED procedure of SPSS. The expression levels of miR-26a were 6.64 folds higher on D16 in pregnant compared to non-pregnant multiparous cows (p < .05). On D8 and D16, miR-26a expression levels were found higher 13 folds in pregnant compared to non-pregnant nulliparous cows (p < .05). Additionally, miR-26a expressions were higher 5.42 folds (p < .05) on D8, 7.19 folds higher (p < .01) on D16 in pregnant nulliparous and multiparous cows, and were 6.30 folds higher (p < .001) on D8 and D16 according to non-pregnant animals. PIF levels were greater in pregnant animals (p < .05). Analyzing miR-26a on D8 might be considered as sufficient in nulliparous cows. Pregnancy detection in multiparous cows can be made on the 16th day with this method. Furthermore, PIF evaluations may be sufficient on D10 in multiparous cows. Besides, PIF levels and miR-26a expression levels might be used safely in field conditions and clinical applications.

The presence of virulence factor genes downregulates uterine AQP3 and alters glutathione peroxidase activity and uterine histopathology in canine pyometra.

Present study was designed to evaluate the role of virulence factor genes (papG, cnf1 and hylA) in the pathogenesis of canine pyometra. Antimicrobial susceptibility test and detection of virulence genes were performed Escherichia coli (E. coli) detected in uterine swab samples. Animals were divided into two groups based on the presence (VF+, n:14) or absence (VF-, n:7) of the virulence factor genes papG, cnf1 and hylA. Blood and tissue glutathione peroxidase activity, uterine histopathologic analysis and AQP3, ESR1, PGR, OXTR gene expressions were determined in both groups. Statistical analyses were performed using Stata version 15.1. All E. coli isolates were susceptible to amikacin, whereas resistant to ampicillin, amoxicillin/clavulanic acid and lincomycin. None of the isolates were susceptible to cefotaxime. E. coli isolates had at least one virulence gene. The most prevalent gene was fimH (100%), followed by fyuA (95.8%), usp (83.3%), sfa (75%), cnf1 and hlyA (70.8%) genes. Blood GPx activity was greater in VF+ animals. On the other hand, uterine tissue GPx activity was lower in VF+ group compared to the control group. Expression levels of AQP3 were upregulated more than fivefold in VF-dogs compared to the control group. In addition, AQP3 expression levels were found approximately threefold higher in VF (-) than VF (+) group (p < .05). Varying degree of inflammation noted for all animals with pyometra, but the presence of bacteria noted only in VF+ animals. In conclusion, the presence of virulence factor genes does not play a role in the histopathological degree of inflammation, the presence of bacteria was found to vary. Serum GPx activity increased in VF+ animals. While the hormone receptor expressions were similar, AQP expression was upregulated in the absence of virulence factor genes.

The protective effect of esculetin against aluminium chloride-induced reproductive toxicity in rats.

One of the prominent health problems caused by Aluminium was the decrease in male fertility rates. In the study, the protective effect of Esculetin (ESC) against the reproductive toxicity induced by Aluminium chloride (AlCl3 ) was investigated. For this purpose, AlCl3 was administrated to Wistar Albino rats at a dose of 34 mg/kg and ESC was administrated at a dose of 50 mg/kg for 70 days. It was determined that AlCl3 treatment reduced sperm motility and concentration, increased dead/live rate and abnormal sperm rate. It decreased serum testosterone level, and co-treatment of ESC significantly regulated these values. In the AlCl3 -treated group, MDA level increased and GSH level, GPx and CAT activities decreased compared with those of the control group. However, co-treatment of ESC showed an amelioratory effect on the values except for CAT activity. It was observed that the expression level of NRF-2 increased in the ESC and AlCl3  + ESC groups, and NF-κB increased in the AlCl3 group with the control group. It was determined that Caspase-3 expression decreased, and Bcl-2 expression increased in AlCl3  + ESC group compared to AlCl3 group. It was also determined that AlCl3 -induced tissue injury was significantly prevented by ESC co-treatment.

Effect of essential oil supplementation to diet on meat quality, fatty acid composition, performance parameters and intestinal microbiota of Japanese quails.

The effect of essential oil (EO) supplementation on carcass characteristics of Japanese quails and interactions between ingredients and intestinal morphology were investigated in this study. A total of 250 quails were fed different diet: D1, basal diet (BD); D2, BD plus palmarosa oil (PO; 100 µg/kg diet); D3, BD plus lemon myrtle oil (LMO; 100 µg/kg diet); D4, BD plus α-Tops (mixture of α-terpineol, cineole and terpinene-4-ol; 100 µg/kg diet); and D5, BD plus cyclodextrin. Overall growth performance was determined at multiple time points during 35 days of experiment. Carcass characteristics (fatty acid, pH and colour), intestinal morphology and the expression levels of meat quality-related genes including the insulin-like growth factor (IGF-1), myogenin and avian uncoupling protein (avUCP) were examined at the end of the trial. Additionally, intestinal microbiome of quails was studied by next-generation sequencing-based culture-independent analysis. Although the inclusion of EOs into the diet had no effect on the growth performance of quails and the microbial profile, the significant changes in pH24 and colour (a*) of the quail's breast muscle (p < .05) in the group receiving PO were observed. Additionally, oleic acid content in the breast muscle was significantly higher in the EOs supplemented groups (p < .01). Quails fed the PO supplemented diet had higher villus and relatively rich in oleic acid. The expression levels of IGF-1 and myogenin genes in quail's muscle were not affected, but the expression of avUCP gene was significantly lower in quails fed with LMO and α-Tops (p < .05). The results demonstrated variable effects of these treatments on intestinal morphology. Taken together, dietary inclusion of EOs is found to be beneficial and hence can be recommended for improving the quality of poultry meat.

Evaluation of Emotional Adverse Effects of Antidepressants: A Follow-up Study.

PURPOSE/BACKGROUND: Emotional adverse effects due to antidepressant use may cause difficulties for the clinician in the treatment of depression. In this prospective study, the emotional adverse effects of antidepressants were evaluated in various aspects. METHODS/PROCEDURES: Ninety eight patients diagnosed with major depressive disorder were included in the study. At 2nd, 4th, 8th, 12th, and 16th weeks, patients were assessed with Montgomery-Asberg Depression Rating Scale (MADRS), and the antidepressant dose was increased in patients with less than a 50% reduction at each visit compared with the initial MADRS score. The Oxford Questionnaire on the Emotional Side-effects of Antidepressants (OQESA) was used at the 8th-week and 16th-week visits. FINDINGS/RESULTS: A significant difference is found in the OQESA score at the 8th-week visit compared with the 16th-week assessment (P < 0.001, t = 5.73). There were significant correlations between MADRS scores and OQESA scores both at the 8th (r = 0.346, P = 0.05) and the 16th (r = 0.490, P < 0.001) weeks. In regression analyses, at eighth-week assessment, MADRS score (B = 1.487, P = 0.002) and selective serotonin reuptake inhibitor use (B = 14.014, P = 0.023) had a significantly predicted OQESA score. IMPLICATIONS/CONCLUSIONS: In this study, it is found that, as the rate of remitted patients is increased, OQESA scores get decreased, and furthermore, the OQESA score of the remitted group is statistically low when compared with that of the nonremitted group at the 8th- and 16th-week visits. Oxford Questionnaire on the Emotional Side-effects of Antidepressants and MADRS scores are significantly correlated in all assessments. These results suggest that the score obtained from OQESA may be related not only to the emotional adverse effects of antidepressants but also to the residual symptoms of depression.

Nobiletin attenuates acetaminophen-induced hepatorenal toxicity in rats.

The study aimed to examine the effects of nobiletin on the toxicity model induced with acetaminophen (APAP). For this purpose, 24 adult male rats were equally divided into four groups. The groups were the control group (group 1); dimethyl sulfoxide only, the APAP group (group 2) received a single dose of APAP 1000 mg/kg on the 10th day of experiment; the Nobiletin group (group 3), nobiletin (10 mg/kg) for 10 days; and the APAP + Nobiletin group (group 4), nobiletin (10 mg/kg) for 10 days with a single dose of APAP (1000 mg/kg) administered on the 10th day and the experiment ended after 48 hours. At the end of the study, a significant increase in malondialdehyde, interleukin-1ß (IL-1ß), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) levels and a significant decrease in glutathione levels, glutathione peroxidase activities and nuclear factor erythroid-derived 2-like 2 (Nrf-2) and heme oxygenase-1 (HO-1) expressions were observed with APAP application in liver and kidney tissues. Serum aspartate transaminase (AST), alanine transaminase (ALT), urea, and creatinine levels were also significantly increased in the APAP group. However, nobiletin treatment in group 4 reversed oxidative stress and inflammatory and histopathological signs caused by APAP. It is concluded that nobiletin may be a beneficial substance that confers hepatorenal protection to APAP-induced toxicity via antioxidant and anti-inflammatory mechanisms.

Decreasing glycerol content by co-supplementation of trehalose and taxifolin hydrate in ram semen extender: Microscopic, oxidative stress, and gene expression analyses.

This study aimed to evaluate the comparative effects of taxifolin hydrate and trehalose on the quality of frozen-thawed ram spermatozoa for the first time. Ejaculates collected from six mature rams were pooled, and divided to eight equal aliquots to extend them with different concentrations of glycerol (%5 and %3), taxifolin hydrate (10, 100, and 500 µM), and trehalose (60 mM) as eight groups (G5T0, G5T10, G5T100, G5T500, G3T0, G3T10, G3T100, and G3T500). After freeze-thawing process of cryopreservation, microscopic and oxidative stress parameters, and gene expression levels were investigated for understanding of possible impacts of taxifolin hydrate and trehalose. The study showed that G3T10 resulted in the highest post-thawed viability and mitochondrial activity. Moreover, all extenders with taxifolin hydrate reduced DNA fragmentation in comparison to G5T0, but DNA damage was prevented at the highest rate in presence of G5T10. The level of LPO significantly decreased in the groups G5T500 and G3T100, and the expression levels of NQO1, GCLC, and GSTP1 genes significantly increased in the groups G5T100, G5T500, G3T10, and G3T100 compared to the group G5T0. Finally, co-supplementation of tris-based extender having 3% glycerol with 60 mM trehalose and 10 µM taxifolin hydrate in cryopreservation extender may be recommended to improve the quality of post-thawed ram spermatozoa. However, further in vivo and in vitro studies are suggested to evaluate fertility rates of frozen-thawed ram spermatozoa co-supplemented with trehalose and taxifolin hydrate.

Dietary high calories from sunflower oil, sucrose and fructose sources alters lipogenic genes expression levels in liver and skeletal muscle in rats.

INTRODUCTION AND OBJECTIVES: The objectives of this study were to investigate the underlying mechanism of PPARα, LXRα, ChREBP, and SREBP-1c at the level of gene and protein expression with high-energy diets in liver and skeletal muscle. MATERIALS AND METHODS: Metabolic changes with consumption of high fat (Hfat), high sucrose (Hsuc) and high fructose (Hfru) diets were assessed. Levels of mRNA and protein of PPARα, LXRα, ChREBP, and SREBP-1c were investigated. Body weight changes, histological structure of liver and plasma levels of some parameters were also examined. RESULTS: In Hfru group, body weights were higher than other groups (P<0.05). In liver, LXRα levels of Hsuc and Hfru groups were upregulated as 1.87±0.30 (P<0.05) and 2.01±0.29 (P<0.01). SREBP-1c levels were upregulated as 4.52±1.25 (P<0.05); 4.05±1.11 (P<0.05) and 3.85±1.04 (P<0.05) in Hfat, Hsuc, and Hfru groups, respectively. In skeletal muscle, LXRα and SREBP-1c were upregulated as 1.77±0.30 (P<0.05) and 2.71±0.56 (P<0.05), in the Hfru group. Protein levels of ChREBP (33.92±8.84ng/mg protein (P<0.05)) and SREBP-1c (135.16±15.57ng/mg protein (P<0.001)) in liver were higher in Hfru group. In skeletal muscle, LXRα, ChREBP and SREBP-1c in Hfru group were 6.67±0.60, 7.11±1.29 and 43.17±6.37ng/mg, respectively (P<0.05; P<0.01; P<0.05). The rats in Hfru group had the most damaged livers. CONCLUSION: Besides liver, fructose consumption significantly effects skeletal muscle and leads to weight gain, triggers lipogenesis and metabolic disorders.

Supramolecular GAG-like Self-Assembled Glycopeptide Nanofibers Induce Chondrogenesis and Cartilage Regeneration.

Glycosaminoglycans (GAGs) and glycoproteins are vital components of the extracellular matrix, directing cell proliferation, differentiation, and migration and tissue homeostasis. Here, we demonstrate supramolecular GAG-like glycopeptide nanofibers mimicking bioactive functions of natural hyaluronic acid molecules. Self-assembly of the glycopeptide amphiphile molecules enable organization of glucose residues in close proximity on a nanoscale structure forming a supramolecular GAG-like system. Our in vitro culture results indicated that the glycopeptide nanofibers are recognized through CD44 receptors, and promote chondrogenic differentiation of mesenchymal stem cells. We analyzed the bioactivity of GAG-like glycopeptide nanofibers in chondrogenic differentiation and injury models because hyaluronic acid is a major component of articular cartilage. Capacity of glycopeptide nanofibers on in vivo cartilage regeneration was demonstrated in microfracture treated osteochondral defect healing. The glycopeptide nanofibers act as a cell-instructive synthetic counterpart of hyaluronic acid, and they can be used in stem cell-based cartilage regeneration therapies.

Evaluation and comparison of the effect of hypothermia and ozone on ischemia-reperfusion injury of skeletal muscle in rats.

BACKGROUND: Tourniquet-induced ischemia-reperfusion, which affects local and distant organs, is very common in orthopedic surgery. Hypothermia is used in traumatic tissue during ischemic period commonly. Ozone (O3) has been recommended as a novel therapeutic agent in various medical conditions. The objective of the study was to evaluate and compare the effect of hypothermia (H) and O3 on ischemia-reperfusion injury of skeletal muscle in rats by measuring oxidative parameters and inducible nitric oxide synthase (iNOS) levels. MATERIALS AND METHODS: Eighteen rats (Wistar albino) were separated into five groups randomly (sham, IR, IR + H, IR + O3, IR + H + O3; n = 6). The lower right extremity of all rats was subjected to 2 h of ischemia and 22 h of reperfusion clamping the common iliac artery and using the rubber-band technique at the level of the lesser trochanter under general anesthesia. Two hours of hypothermia were applied during the first 2 h of reperfusion in two groups. O3 was applied in two groups. All rats were sacrificed after the IR period with high dose of anesthesia. The tibialis anterior muscle and blood were saved. Levels of superoxide dismutase, glutathione peroxidase, MDA, NOx, and interleukin-1ß were measured in the muscle. Creatinine kinase, lactate dehydrogenase, aspartate aminotransferase, urea, creatinine, and electrolytes were measured in serum. Immunohistochemical iNOS staining was performed on muscle samples. RESULTS: The levels of MDA, NOx, and interleukin-1ß in muscle were raised in the IR group compared with those in the sham group. The same parameters were lower in the groups of IR + H, IR + O3, and IR + H + O3 compared with those in the IR group. Superoxide dismutase and glutathione peroxidase activities in muscle were lower in the IR group compared with those in the sham group; however, same parameters were higher in the groups of IR + H, IR + O3, and IR + H + O3 compared with those in the IR group. Score and intensity of iNOS staining in skeletal muscle in the IR group was increased compared with that in the sham group and decreased in the groups of IR + H, IR + O3, and IR + H + O3 compared with that in the IR group. Levels of creatinine kinase, aspartate aminotransferase, and K in the three treatment groups decreased compared with those in the IR group. CONCLUSIONS: These findings showed that hypothermia, which has more affect, and O3 decreased the tourniquet-induced IR injury in the rat's muscle-skeletal system by reducing the levels of oxidative and nitrosative stress parameters and enhancing antioxidant enzymes. Hypothermia and O3 had no synergistic effect. Hypothermic reperfusion and O3 preconditioning might be beneficial in skeletal muscle IR injury-associated tourniquet.

Considering potential roles of selected MicroRNAs in evaluating subclinical mastitis and Milk quality in California mastitis test (+) and infected bovine milk.

This study investigates the relationships between subclinical mastitis and milk quality with selected microRNAs in cow milk. California Mastitis Test (CMT)-positive (n = 20) and negative (n = 20) samples were compared (Experiment I). Additionally, samples with CMT-positive but microbiological-negative, as well as positive for only Staphylococcus subspecies (Staph spp.) and only Streptococcus subspecies (Strep spp.) were examined (Experiment II). Four groups were formed in Experiment II: Group I (CMT and microbiological-negative) (n = 20), Group II (CMT-positive but microbiological-negative) (n = 10), Group III (Staph spp.) (n = 5), Group IV (Strep spp.) (n = 5). While electrical conductivity, somatic cell count (SCC), malondialdehyde (MDA) increased, miR-27a-3p and miR-223 upregulated and miR-125b downregulated in the CMT-positive group in Experiment I. SCC and MDA were higher in CMT-positive groups. miR-27a-3p and miR-223 upregulated in Groups III and IV. While miR-155 is upregulated, miR-125b downregulated in Group IV. Milk fat is positively correlated with miR-148a and miR-223. As miR-27a-3p positively correlated with SCC and MDA, miR-125b negatively correlated with electrical conductivity and SCC. miR-148a and MDA were positively correlated. miR-155 was correlated with fat-free dry matter, protein, lactose, and freezing point. miR-223 was positively correlated with SCC and miR-148a. Results particularly highlight miR-27a-3p and miR-223 as potential biomarkers in subclinical mastitis, especially those caused by Staph spp. and Strep spp., while miR-148a, miR-155, and miR-223 stand out in determining milk quality.

Expression patterns and distribution of aquaporin water channels in cervix as a possible mechanism for cervical patency in bitches affected by pyometra.

Pyometra is a life-threatening disease, the severity of which depends on cervical patency status. This study investigated cervical inflammation status as well as the expression patterns and localization of aquaporin (AQP1, AQP2, AQP3, AQP5, and AQP9), and hormone receptors in cervical tissue that influences canine pyometra. Of the 36 animals enrolled in the study, 24 were diagnosed with pyometra and separated into two groups: open cervix pyometra and close cervix pyometra, while 12 healthy animals presented for elective ovariohysterectomies were allocated into the control group. Surgical treatment was performed for treatment of pyometra. After each operation, cervix samples were collected and analyzed for AQP and hormone receptor expression patterns determined by qPCR and protein expression by means of immunohistochemistry. Blood samples were also collected to determine serum progesterone concentrations. AQP9 expression was downregulated approximately 3-fold while and PGR expression was downregulated more than 2 fold in both pyometra groups compared to the control group. AQP3 and AQP5 gene expression levels were upregulated more than 3 fold in the open-cervix pyometra group than the closed-cervix pyometra group (P < 0.05). This is the first study to describe the expression patterns and immunolocalization of AQPs in canine cervical tissue based on pyometra patency status and to report AQP3 and AQP5 expression in cervical tissue linked to cervical patency.

Expression patterns of inflammatory and oxidative stress-related genes in the uterine and ovarian tissues of dogs diagnosed with pyometra based on cervical patency status.

This study aimed to investigate the expression patterns of genes associated with inflammation and oxidative stress in ovarian and uterine tissues of dogs with pyometra, categorized by cervical status (open cervix or closed cervix), which influences disease severity. The control group comprised healthy animals undergoing elective ovariohysterectomy. Tissue inflammatory gene expression and Malondialdehyde (MDA) levels were determined while microbial and histopathological examinations were conducted, along with immunohistochemical evaluations. In the closed-cervix group, uterine TNF and IL6 were upregulated approximately 10-fold while IL10 was upregulated nearly 5-fold. TNF expression differed remarkably between the pyometra groups. In the closed-cervix group, PTGS2 and HMOX1 were upregulated approximately 5-fold whereas NFE2L2 expression was downregulated. The closed-cervix group also had the highest uterine MDA levels. Regarding ovarian tissue, MDA levels were higher in the closed-cervix group than in the open-cervix group while IL10 expression was lower in the closed-cervix group than the open-cervix group. In the closed-cervix group, NFE2L2 was downregulated whereas HMOX1 was upregulated. Uterine TNF levels were positively correlated with IL6, IL10, PTGS2, and HMOX1, but negatively correlated with NFE2L2. IL6 was positively correlated with IL10, PTGS2, and HMOX1. NFE2L2 was negatively correlated with IL6 and HMOX1. IL10 was positively correlated with PTGS2 and HMOX1. MDA was positively correlated with TNF, IL6, IL10, PTGS2, NFE2L2, and HMOX1. TNF levels were positively correlated with ovarian PTGS2, and with IL6 and NFE2L2. MDA was positively correlated with PTGS2 and HMOX1. MDA could be an important biomarker for understanding the severity of pyometra. Moreover, TNF expression and its relationships with various studied parameters such as IL10 may contribute to treatment and prognostic biomarker studies in closed-cervix pyometra pathology.

The effects of the feeding duration of propylene glycol on major meat quality parameters and substantial proteins in the muscle of Akkaraman lambs.

In this study, the effects of propylene glycol (PG) on meat quality and molecular pathways related to energy metabolism in longissimus lumborum muscle on lambs were evaluated. Seventy-two lambs were divided into three groups consisting of 60th, 90th, and 120th of slaughter days. The dosage of the PG and slaughter days were the variables used in the study. Eight animals were slaughtered from each group on each day. The meat quality parameters (e.g., pH, protein, fatty acid profile) and IGF-1, IGFBP4, and DGAT1 (i.e., mRNA and protein levels) were evaluated. The pH 45 min post-slaughter was higher in PG groups on 120th day. On the 4th day after slaughter, the b value was the lowest in the PG3, while 7th day after slaughter it was highest in Con and PG3 on 90th day. The total n3 and n6 were lowest and the NV was highest on 120th day. The IGFBP4 was upregulated in the PG groups on all of the slaughter days. The DGAT1 was upregulated in the PG3 on the 90th day. The IGF-1, DGAT1, IGFBP4 protein levels were found to have increased in the PG3 on 90th day. The IGFBP4 was found to have decreased in the PG3 on 120th day. According to the results of the study, the oral administration of the PG at the 3 mL/kg live weight0.75 for at least 120 days may have positive effects on meat quality in lambs through the IGF-1, DGAT1, and IGFBP4 genes and the proteins encoded by these genes.

Transfer learning of an ensemble of DNNs for SSVEP BCI spellers without user-specific training.

Objective.Steady-state visually evoked potentials (SSVEPs), measured with electroencephalogram (EEG), yield decent information transfer rates (ITRs) in brain-computer interface (BCI) spellers. However, the current high performing SSVEP BCI spellers in the literature require an initial lengthy and tiring user-specific training for each new user for system adaptation, including data collection with EEG experiments, algorithm training and calibration (all are before the actual use of the system). This impedes the widespread use of BCIs. To ensure practicality, we propose a novel target identification method based on an ensemble of deep neural networks (DNNs), which does not require any sort of user-specific training.Approach.We exploit already-existing literature datasets from participants of previously conducted EEG experiments to train a global target identifier DNN first, which is then fine-tuned to each participant. We transfer this ensemble of fine-tuned DNNs to the new user instance, determine thekmost representative DNNs according to the participants' statistical similarities to the new user, and predict the target character through a weighted combination of the ensemble predictions.Main results.The proposed method significantly outperforms all the state-of-the-art alternatives for all stimulation durations in [0.2-1.0] s on two large-scale benchmark and BETA datasets, and achieves impressive 155.51 bits/min and 114.64 bits/min ITRs. Code is available for reproducibility:https://github.com/osmanberke/Ensemble-of-DNNs.Significance.Our Ensemble-DNN method has the potential to promote the practical widespread deployment of BCI spellers in daily lives as we provide the highest performance while enabling the immediate system use without any user-specific training.

The Matrix Metalloproteinase Inhibitor Batimastat Reduces Epidural Fibrosis After Laminectomy in Rats.

AIM: To investigate the efficacy of locally applied batimastat after laminectomy in preventing postoperative epidural fibrosis. MATERIAL AND METHODS: Thirty-two Wistar albino male rats weighing 200?250 g were used. The rats were assigned to four different groups (I-Control group, II-sham group, III-Laminectomy+Batimastat group, and IV-Laminectomy+SpongostanTM group). The rats were euthanized 28 days after surgery before TNF-?, IL6, IL-1?, IL10, TGF-?1, and MMP9 gene expression levels of tissue in the surgical area were determined with qPCR. TNF-?, IL6, and IL10 protein levels were also measured in both tissue and plasma. In addition, the surgical area was evaluated by histopathological and immunohistochemical methods. RESULTS: TNF-?, IL6, and IL-1? gene expression levels were higher in the batimastat group than in the control group. Whereas IL10 gene expression levels increased about two-fold in the sham and SpongostanTM groups, in the batimastat group, it was similar to that in the control group. TGF-?1 gene expression was three-fold higher in the sham group but was similar to that in the control group in both batimastat and SpongostanTM groups. MMP9 gene expression levels significantly decreased only in the batimastat group. In addition, fibrosis score, fibroblast cell count, inflammatory cell count, and CD105 expression decreased in the batimastat group relative to the control. CONCLUSION: Molecular and pathological examination results suggested that batimastat is an effective agent in reducing the occurrence of epidural fibrosis after laminectomy.

The thermographic monitoring in early pregnancy detection in Holstein cows and heifers.

This study aimed to conduct a comprehensive investigation to demonstrate early pregnancy detection using thermography in heifers and cows. A total of 60 heifers (n: 30) and cows (n: 30) were divided into two groups as pregnant (n: 15 heifers, n: 15 cows) and non-pregnant (n: 15 heifers, n: 15 cows) according to the day 28 of gestation. Thermographic images were taken from the vulvar and anal regions on alternate days from D0 to D20. Blood samples were collected to determine estrogen and progesterone concentrations. The mean temperature difference between the anal and vulvar regions (ΔT °C) was used in the statistical analyses. Based on the hormonal profiles, no abnormalities were observed for follicular waves or luteal profiles in heifers and cows. The ΔT °C values between heifers and cows and between days were statistically significant (P < 0.001). In thermographic analyses, the differences observed in other main effects and interactions of the group, sampling time, and pregnancy were not statistically significant (P > 0.05). In receiver operating characteristic (ROC) analyses, it was concluded that the ΔT °C value of ≤ 2.9 °C (100% Se - 61.9% Sp) was highly correlated with pregnancy diagnosis in cows on day six after artificial insemination (AI) (P < 0.001). In conclusion, it was determined that the clinical application of thermography can be used for the detection of pregnancy on day six after AI in cows. However, further studies are needed to determine heifers' thermographic characteristics and profiles.

Protective and Therapeutic Effects of Bovine Amniotic Fluids Collected in Different Trimesters on the Epidural Fibrosis After Experimental Laminectomy in Rats.

BACKGROUND: The aim of this study was to investigate the protective and therapeutic effects of bovine amniotic fluid (BAF) on the inhibition of epidural fibrosis (EF) after experimental laminectomy. METHODS: Forty female Sprague Dawley rats were used. The amniotic fluids were collected from each trimester of a pregnant cow. The rats were divided into 5 groups. Whereas no laminectomy was applied to the control group, animals in the sham group underwent laminectomy. Laminectomy was performed in the animals in other groups and the operation area was closed by dripping 1 mL of BAF collected in 3 trimesters of pregnancy. Animals were killed 28 days after the operation. RESULTS: Compared with control, VEGF gene expression levels were downregulated approximately 5-fold in BAF-2. Whereas IL-6 was upregulated approximately 8-fold in the sham, it was downregulated 5-fold and 3-fold in BAF-1 and BAF-2, respectively. There was downregulation in BAF-2 and BAF-3 in terms of CD105 gene expression levels. TGFß1 was upregulated approximately 2-fold in the sham group and downregulated in BAF-1 and BAF-2. Although histopathologic alterations including EF grade and fibroblast cell density were found to increase in the sham group, all BAF treatment decreased those of alterations. The highest CD105 immunoreactivity was detected in the sham group. All BAF treatment markedly aggravated fibrosis via decreasing CD105 immunoreactivity. In terms of grading parameters, almost the closest grades to the control were determined in the BAF-2. BAF collected in the second trimester is most effective in healing of scar tissue and preventing fibrosis via decreasing microvessel and fibroblast densities. CONCLUSIONS: The results indicate that BAF may be used as a potential protective agent to prevent EF.

Sustained inhibition of CC-chemokine receptor-2 via intraarticular deposition of polymeric microplates in post-traumatic osteoarthritis.

Posttraumatic osteoarthritis (PTOA) is mostly treated via corticosteroid administration, and total joint arthroplasty continues to be the sole effective intervention in severe conditions. To assess the therapeutic potential of CCR2 targeting in PTOA, we used biodegradable microplates (µPLs) to achieve a slow and sustained intraarticular release of the CCR2 inhibitor RS504393 into injured knees and followed joint damage during disease progression. RS504393-loaded µPLs (RS-µPLs) were fabricated via a template-replica molding technique. A mixture of poly(lactic-co-glycolic acid) (PLGA) and RS504393 was deposited into 20 × 10 µm (length × height) wells in a polyvinyl alcohol (PVA) square-patterned template. After physicochemical and toxicological characterizations, the RS504393 release profile from µPL was assessed in PBS buffer. C57BL/6 J male mice were subjected to destabilization of the medial meniscus (DMM)/sham surgery, and RS-µPLs (1 mg/kg) were administered intraarticularly 1 week postsurgery. Administrations were repeated at 4 and 7 weeks post-DMM. Drug free-µPLs (DF-µPLs) and saline injections were performed as controls. Mice were euthanized at 4 and 10 weeks post-DMM, corresponding to the early and severe PTOA stages, respectively. Knees were evaluated for cartilage structure score (ACS, H&E), matrix loss (safranin O score), osteophyte formation and maturation from cartilage to bone (cartilage quantification), and subchondral plate thickness. The RS-µPL architecture ensured the sustained release of CCR2 inhibitors over several weeks, with ~ 20% of RS504393 still available at 21 days. This prolonged release improved cartilage structure and reduced bone damage and synovial hyperplasia at both PTOA stages. Extracellular matrix loss was also attenuated, although with less efficacy. The results indicate that local sustained delivery is needed to optimize CCR2-targeted therapies.

The role of nutritional-immunological indices in estimating serum LPS and antioxidant enzyme activity and sepsis status in female dogs with pyometra caused by E. coli.

The aim of this study was to diagnose pyometra and related sepsis status using cost-effective nutritional-immunological indices, antioxidants, and toxin levels in dogs and to investigate the utility of the indices in predicting toxin and antioxidant status. A total of 29 dogs were enrolled into the present study. Among these, 9 female dogs in their diestrus stages, were allocated for elective ovariohysterectomy. The pyometra group was also separated into two subgroups as Sepsis (+) and Sepsis (-). Blood samples were collected into two tubes containing EDTA for hematological analysis; without anticoagulant for serum progesterone, LPS concentration, and antioxidant levels at the time of diagnosis. Bacteriological and tissue samples of the uterus were collected after the ovariohysterectomy. Antioxidant activity, progesterone, and toxin concentration were determined by using commercial ELISA kits. Statistical analyses were performed using Stata version 16.1 and MedCalc 16 statistical software. Receiver operating characteristics curves were used for the threshold for evaluating pyometra and sepsis status. Pairwise comparisons were carried out of the area under the curve (AUC) for thresholds of nutritional immunologic indices (hemoglobin, albumin, lymphocyte, platelet (HALP) score; prognostic nutritional index (PNI); Albumin hemoglobin index (AHI)), serum LPS and antioxidant activity. Linear regression model was used for the estimation of serum LPS and antioxidant activity by using indices. Mean serum progesterone, LPS concentrations, and Nitric Oxide (NO) production were greater, while serum superoxide dismutase (SOD), tissue SOD, and glutathione peroxidase (GPx) activities were lower in dogs with pyometra. All nutritional-immunologic indices were lower in pyometra cases. Nutritional-immunologic indices (AUC of HALP:0.759; PNI:0.981; AHI 0.994), NO (AUC: 0.787) and SOD (AUC: 0.784) levels were useful for pyometra diagnosis. AHI and LPS were useful for the determination of sepsis status with the AUC values of 0.850 and 0.740, respectively. While AHI was useful for the estimation of serum LPS and NO concentration (p < 0.001), PNI was useful for serum SOD concentration (p = 0.003). In conclusion, PNI, HALP and AHI can be used in the diagnosis of pyometra, however, only AHI and LPS levels can be used in the diagnosis of sepsis. SOD and NO can be used to determine pyometra but have no effect on determining sepsis status. Additionally, the estimation of the levels of serum LPS, NO, and SOD activities can be done using the AHI and PNI values.