Effect of direct therapeutic ultrasound exposure of ovaries on histopathology, inflammatory response, and oxidative stress in dogs.

BACKGROUND: This research was designed to evaluate the effects of therapeutic ultrasound waves on ovarian germinal tissue and inflammatory cytokines (interleukin-6 (IL-6), IL1ß, tumor necrosis factor-α (TNF-α)), acute phase proteins (serum amyloid A (SAA), C reactive protein (CRP)) and oxidative stress (total antioxidant capacity (TAC), and malondialdehyde (MDA)) in dogs. Twenty-six clinically healthy adult mix-breed female dogs were aligned into three groups. Laparotomy was performed in control (n = 6) and treatment (T5, n = 10; T10, n = 10) groups. The ultrasonic exposure of ovaries in treatment groups was performed during laparotomy by round motions of the therapeutic ultrasonic transducer on both ovaries (1 MHz frequency, 1.5 W/cm2) for 5 min in the T5 group and for 10 min in the T10 group. Blood samples were collected from the jugular vein into a plain glass tube on days 0 (before laparotomy), 3, 6, and 9 after surgery. All control and treatment groups' dogs were ovariectomized for histological evaluation on day 60 after laparotomy or laparotomy + ultrasound exposure. RESULTS: Direct exposure of ovaries with therapeutic ultrasound waves induced inflammation and oxidative stress comparison with the control group. Histopathological evaluation of treated ovaries with ultrasound waves indicated a decreased number of primordial follicles (ovarian reserve) and oocyte preservation scores compared with ovaries in the control group. CONCLUSIONS: These changes may cause subfertility in the long term. It seems that inflammatory response and oxidative stress are factors in the permanent damage of ovarian tissue.

The effects of melatonin on the concentrations of inflammatory cytokines and proteins, serotonin, cortisol and melatonin in ovariohysterectomised female dogs.

BACKGROUND: Ovariohysterectomy (OHE) induces inflammation and stress in female dogs. The anti-inflammatory effects of melatonin have been reported in several studies. OBJECTIVES: The goal of this study was to assess the effects of melatonin on the concentrations of melatonin, cortisol, serotonin, α-1-acid glycoprotein (AGP), serum amyloid A (SAA), c-reactive protein (CRP), interleukin-10 (IL-10), interleukin-8 (IL-8), interleukin-1ß (IL-1ß) and tumour necrosis factor-α (TNF-α) before and after OHE. METHODS: The total number of animals was 25 and aligned in 5 groups. Fifteen dogs were divided into three groups (n = 5): melatonin, melatonin+anaesthesia and melatonin+OHE and received melatonin (0.3 mg/kg, p.o.) on days -1, 0, 1, 2 and 3. Ten dogs were assigned to the control and OHE groups (n = 5) without melatonin treatment. OHE and anaesthesia were performed on day 0. Blood samples were obtained via jugular vein on days -1, 1, 3 and 5. RESULTS: Melatonin and serotonin concentrations significantly increased in the melatonin, melatonin+OHE and melatonin+anaesthesia groups compared with the control group, while cortisol concentration decreased in the melatonin+OHE group compared with the OHE group. The concentrations of acute-phase proteins (APPs) and inflammatory cytokines significantly increased after OHE. The CRP, SAA and IL-10 concentrations decreased significantly in the melatonin+OHE group compared with the OHE group. The concentrations of cortisol, APPs and proinflammatory cytokines increased significantly in the melatonin+anaesthesia group compared with the melatonin group. CONCLUSIONS: The oral administration of melatonin before and after OHE help controlling the high levels of inflammatory APPs, cytokines and cortisol induced by OHE in female dogs.

Effects of short-term administration of melatonin before gonadectomy on oxidative stress, cortisol and sex hormones in male dogs.

The aim of this study was to investigate gonadectomy stress, steroid hormones and serotonin in male dogs treated with melatonin before gonadectomy. Twenty-five mixed breed adult dogs were divided into five equal groups. The melatonin and melatonin + gonadectomized groups received melatonin treatment (3 mg/10 Kg, PO, TID) the day before gonadectomy; the gonadectomized and anaesthesia groups did not receive melatonin; and the control group just received the melatonin vehicle. Blood sampling was performed before melatonin administration (day -1) and on days 0 (gonadectomy), 1, 3 and 6 after gonadectomy. Superoxide dismutase and glutathione peroxidase concentrations decreased significantly in gonadectomized dogs compared with dogs treated with melatonin before gonadectomy and intact dogs. Gonadectomy led to a significant decrease in catalase concentration in gonadectomized dogs compared with other study groups. Malondialdehyde levels increased significantly in gonadectomized dogs compared with other groups. Melatonin administration before gonadectomy led to decreased malondialdehyde concentration in gonadectomized and intact dogs compared to the control group. Cortisol concentration increased significantly in gonadectomized dogs compared to the control dogs. Serotonin levels decreased in gonadectomized dogs, but melatonin treatment increased serotonin concentration in gonadectomized and intact dogs. Melatonin treatment before gonadectomy suppressed oxidative stress and the cortisol but increased serotonin level.

The effects of melatonin treatment on oxidative stress induced by ovariohysterectomy in dogs.

BACKGROUND: As one of the most common surgeries performed in veterinary medicine, ovariohysterectomy (OHE) can induce oxidative stress in dogs. The antioxidant properties of melatonin have been confirmed in various studies. This study aimed to investigate the effects of melatonin administration on oxidative stress in dogs before and after OHE. In this study, 25 mature female intact dogs were selected and randomly divided into five equal groups: Melatonin (melatonin, no surgery), OHE (no melatonin, surgery), OHE + melatonin (melatonin, surgery), anesthesia+melatonin (melatonin, sham surgery), and control (no melatonin, no surgery) groups. Melatonin (0.3 mg/Kg/day, p.o.) was administrated to the dogs in the melatonin, OHE + melatonin, and anesthesia+melatonin groups on days - 1, 0, 1, 2, and 3 (day 0 = OHE). Blood sampling was performed on days - 1, 1, 3, and 5 of the study. Blood samples were immediately transferred to the laboratory and sera were separated and stored at - 20 °C. Superoxide dismutase (SOD), glutathione peroxidase (GPX), catalase (CAT) and malondialdehyde (MDA) concentrations were measured with commercial kits. RESULTS: The levels of SOD, GPX and CAT were significantly higher in the melatonin and anesthesia+melatonin groups compared to those of the control group at days 3 and 5. The level of antioxidant enzymes significantly decreased in the OHE group compared to that of other groups at days 3 and 5. The administration of melatonin increased the level of antioxidant enzymes in ovariohysterectomized dogs. Ovariohysterectomy significantly increased the concentration of MDA in comparison to that of other groups at day 3. Melatonin administration significantly decreased the level of MDA in melatonin, anesthetized, and ovariohysterectomized dogs at day 3. CONCLUSIONS: Administration of melatonin on day - 1, 0, 1, 2 and 3 modulate the oxidative stress induced by OHE in dogs by increasing antioxidant enzymes concentration and decreasing MDA levels.

Echotexture Analysis of Prostate Parenchyma for Detection of Benign Prostatic Hyperplasia in Dogs.

Ultrasonography is one of the most common methods for the diagnosis of prostate disorders, such as benign prostatic hyperplasia (BPH), in dogs. Changes in the echotexture are one of the indicators used to diagnose prostate disorders. The purpose of this study was to investigate the changes occurred in the dogs' prostate echotexture during the induction of BPH using image analysis. Twenty sexually mature male intact mixed-breed dogs were selected and divided randomly into control and BPH-induced groups. BPH was induced using testosterone and estrogen injections for 63 days. The ultrasound imaging of the dogs' prostate was performed during the induction of BPH on days 0, 21, 42, and 63. The echotexture of the prostate parenchyma was analyzed using the Image J software. Then, the changes in the echotexture and its correlation and linear regression with the prostate volume and canine prostate specific esterase (CPSE) concentration were evaluated by statistical tests. The prostate parenchyma echotexture did not show any significant changes during the induction of BPH and in comparison with that of the control group. While prostate volume and CPSE concentration increased significantly, indicating that BPH was induced in the dogs. There was no significant correlation and linear regression between the prostate echotexture and prostate volume or between the CPSE concentration and prostate echotexture. According to the results, the alteration in the prostate parenchymal echotexture did not occur in the early stages of induced BPH, but significant changes occurred in the prostate volume and CPSE concentration during those early stages.

Direct endoscopic lavage and biopsy sampling and evaluation of uterine microflora in various stages of the canine estrous cycle.

The microbial population of the uterus fluctuates during the estrous cycle. Microflora of uterus may affect the establishment and maintenance of pregnancy in bitches. The endoscopic samples obtained from the vagina and uterus of 20 female adult mixedbreed dogs. The uterine lavage samples were prepared for cytology, bacterial (aerobic and anaerobic) and fungal cultures. Uterine tissue samples were evaluated for the presence of E. coli by the polymerase chain reaction. The pure growth of bacteria was observed in seven plates out of the nineteen cultured samples (36.84%) and five Gram-negative and two Gram-positive bacteria were detected. The highest number of isolated bacteria was related to the samples of the diestrus and anestrus stages of the estrous cycle, while the lowest number of bacteria was observed in the samples of the estrous stage. Moreover, Citrobacter spp. was the most frequent group of isolated bacteria. The neutrophils were detected in the cytology of uterine samples. The fungi growth was observed in three uterine samples. Cladosporium and Penicillium isolated from the samples were related to the estrus stage, and yeast was grown in diestrus samples. The 16srRNA gene existed in all of the estrous uterine samples in which the bacterial culture was negative. However, the presence of this gene was proven in two samples (33.30%) of negative bacterial culture samples from the diestrus and anestrus stages. In conclusion, the normal bitches' uteri were infected with various bacteria in estrus, diestrus and anestrus stages of the estrous cycle, and it could coincide with the fungi infection.

Changes in the Serum Prostatic Biomarkers During the Treatment of Benign Prostatic Hyperplasia with a 5alpha-reductase Inhibitor: Finasteride.

The monitoring of serum prostatic biomarkers during the treatment will help clinicians to know the statement of the response to finasteride in dogs affected by benign prostatic hyperplasia (BPH). The present study was aimed to assess changes in the serum canine prostate-specific esterase (CPSE), prostate-specific antigen (PSA), prostatic acid phosphatase (PAP), testosterone, dihydrotestosterone (DHT) and prostate volume evaluation using ultrasonographic examination during the treatment with finasteride in BPH-induced dogs. Twenty dogs were divided into 4 groups (n = 5): BPH + finasteride group, dogs which were induced for BPH and received oral finasteride once daily for 1 month; BPH group, dogs which were induced for BPH and received placebo; finasteride group, normal dogs which received finasteride; and normal group, normal intact dogs which did not receive treatment. Blood sampling and ultrasonography examination were performed on days 0, 14, and 28. The administration of finasteride led to a significant decrease in the concentration of the prostate-specific biomarkers (PSA, CPSE), DHT, testosterone, and the volume of the prostate in BPH + finasteride group compared with the BPH group during 1 month. Interestingly, the PAP concentration did not change in the BPH-induced dogs and in dogs treated with finasteride. It seems that the monitoring of serum PSA and CPSE levels and ultrasonographic examination of the prostate are useful methods for following up the response to finasteride treatment in dogs affected by BPH.

Treatment of experimentally induced benign prostatic hyperplasia with Tadalafil and castration in dogs.

New methods are being developed for the treatment of benign prostatic hyperplasia (BPH) in dogs. The aim of the present study was to evaluate the effects of Tadalafil on the treatment of experimentally induced BPH in dogs. Twenty-five adult intact male dogs were randomly divided into five groups (n = 5): normal group; dogs induced with BPH and treated with Tadalafil (5 mg/day p.o.); dogs which received Tadalafil (5 mg/day p.o.); dogs induced with BPH and treated with castration; and dogs induced with BPH. For 4 sequential weeks, the hematologic and prostate-specific factors (dihydrotestosterone (DHT), serum prostate-specific antigen (PSA), serum prostatic acid phosphatase (PAP), and canine prostatic specific esterase (CPSE)) were measured. Significant differences were observed in the level of PSA, CPSE, and PAP concentration between the normal vs. BPH-Tadalafil, BPH-castrated, and BPH groups. Treating BPH-induced dogs with Tadalafil or castration significantly declined the serum PSA, CPSE, and PAP levels compared to those of the untreated BPH-induced group. The treatment of normal dogs with Tadalafil did not affect prostate-specific biomarkers in comparison with normal dogs. In conclusion, and according to the prostatic indices, it could be stated that Tadalafil, compared with castration, could be used for the treatment of BPH in dogs.

Changes in specific serum biomarkers during the induction of prostatic hyperplasia in dogs.

BACKGROUND: Prostatic hyperplasia (PH) is one of the most important disorders in intact dogs. In this study, we aimed to induce PH experimentally using the combination of testosterone and estrogen and evaluate important factors associated with this disease. RESULTS: The results showed that in the induction group, prostate volume and prostate specific antigen (PSA) concentration increased significantly on day 21 onwards compared to those of the control group. Canine prostatic specific esterase (CPSE) and dihydrotestosterone (DHT) concentrations increased significantly on day 42 onwards while the testosterone levels increased on day 63. In addition, prostatic acid phosphatase (PAP) concentration did not change significantly in the control and induction groups. Biochemistry profiles and hematologic factors were measured for monitoring the function of liver and kidney, and there were no adverse effects following the induction of PH. CONCLUSIONS: It seems that testosterone and estrogen administration led to prostatic hyperplasia during 2 months. Investigating the size of the prostate, accompanied by prostate markers including CPSE, PSA, DHT, and testosterone, is helpful for the PH diagnosis. However, further studies should be carried out on PAP.

MicroRNA expression in infertile men: its alterations and effects.

Infertility is an important reproductive health problem, and male infertility is especially important in more than half of infertility cases. Due to the importance of genetic factors in this condition, analysis of semen alone is not enough to recognize men with idiopathic infertility. A molecular non-invasive investigation is necessary to gain valuable information. Currently, microRNAs (miRNAs) are being used as non-invasive diagnostic biomarkers. miRNAs, single-stranded non-coding RNA molecules, act as post-transcriptional gene silencing regulators either by inhibition or repression of translation. Changes in the regulation of miRNAs have been investigated in several different types of male infertility, therefore the biological role of miRNA and gene targets has been defined. The purpose of this study was to review recent research on the altered expression of miRNA in semen, sperm, and testicular biopsy samples in infertile males with different types of unexplained infertility. Changes in miRNA regulation were investigated using microarray and the miRNA levels were confirmed by real-time qRT-PCR. This review explains why creating a non-invasive diagnostic method for male infertility is necessary and how changes in miRNA expression can be used as new diagnostic biomarkers in patients with differing spermatogenic and histopathologic injury.

Changes in the oxidative stress factors and inflammatory proteins following the treatment of BPH-induced dogs with an anti-proliferative agent called tadalafil.

BACKGROUND: Finding a medical treatment which can combat cell proliferation and relax smooth muscles in canine benign prostatic hyperplasia (BPH) appears to be imperative. AIMS: This study aimed to evaluate the oxidative stress and inflammatory proteins following the treatment of dogs induced for BPH with an anti-proliferative agent called tadalafil. MATERIALS AND METHODS: Twenty-five adult intact male dogs were randomly designated into five groups (n = 5): Control group was not induced for BPH and not treated with tadalafil; dogs induced for BPH by testosterone enanthate and estradiol benzoate and treated with tadalafil (5 mg/day P.O.); dogs which received tadalafil (5 mg/day P.O.); dogs induced for BPH and treated with castration; and dogs induced for BPH. Oxidative stress factors (glutathione peroxidase [GPX], superoxide dismutase [SOD], catalase) and inflammatory proteins (haptoglobin, serum amyloid A [SAA], malondialdehyde [MDA]) were measured in the blood serum for four sequential weeks. RESULTS: Glutathione peroxidase and SOD serum levels declined in dogs in the BPH-induced group compared to those in the control group. Those levels diminished in BPH-induced castrated and tadalafil-treated groups. The changes in the GPX and SOD serum concentrations were not significant between the BPH-induced castrated group and BPH-induced tadalafil-treated group. Moreover, MDA concentration increased slightly in groups with BPH and groups which were castrated. Generally, however, there were no significant differences in the MDA serum concentrations between other groups. Haptoglobin and SAA concentrations increased in BPH-castrated group. Also, the differences in haptoglobin and SAA were not significant between the groups. CONCLUSION: Tadalafil could not control oxidative stress and inflammatory mediators which happened during BPH in dogs.

The Clinical Follow-Up of Estradiol Benzoate Priming During Induction of Estrus With Cabergoline in Dogs.

Induction of estrus is used to improve reproductive efficiency of female dogs. In this study, 11 adult healthy female dogs were selected at anestrus stage. The dogs were assigned to treatment (6 dogs) and control groups (5 dogs). Single dose of estradiol benzoate was injected in treatment group at day 0 (0.01mg/kg, IM). Dogs in both groups received cabergoline (5µg/kg orally) from day 7 to the onset of proestrus. Vaginal cytology and blood samples were taken twice a week during study. Average time to the onset of proestrus was 10.33 ± 4.2 and 15 ± 7.5 days in the treatment and control groups, respectively (P = .08). The differences in time to the onset of estrus phase in the treatment group (14.67 ± 5.9 days) and control group (18.67 ± 10.8 days) were significant (P < .05). The average length of proestrus phase in treatment and control groups was 5.33 ± 2.2 and 8 ± 4.6 days, respectively and their differences were significant (P < .05). Average length of estrus phase in treated dogs with estradiol benzoate was 8.57 ± 3.5 days but it was 8 ± 4.6 days in control group (P > .05). Administration of cabergoline caused significant decrease of prolactin concentration in both groups (P < .01). The difference in serum prolactin concentration between treatment and control was not significant. The effect of cabergoline on serum prolactin concentration was not affected by administration of estradiol benzoate in treatment group (P > .05). As a result, administration of estradiol benzoate 1 week before cabergoline improved induction and synchronization of estrus in dogs.