Late effects of cutaneous 3-methylcholanthrene exposure on DNA damage-related pleiotropic growth factors and oxidative stress markers in mice.

BACKGROUND: Skin is the body's first defence against direct exposure to variety of chemicals. Polycyclic aromatic hydrocarbons such as 3-methylcholanthrene (3-MC) are common in polluted urban air and have a potential of producing harmful effects. Moreover, their late effects can occur months or years after exposure. OBJECTIVES: We aimed to investigate the long-term effects of 3-MC induced dermal toxicity on the expression of markers of apoptosis, pleiotropic cytokines, and oxidative stress and to determine the protective effect of cisplatin. METHODS: Groups were designed as control (group 1), 3-MC applied (group 2) and 3-MC+cisplatin applied mice (group 3). Cutaneous expressions of TGFß, PDGFA, PDGFC, bFGF, PDGFRα, USP28, and Ki67 were evaluated with qPCR. Total oxidant (TOS), total antioxidant (TAS) and oxidative stress index (OSI) values were determined in liver and kidney tissues. RESULTS: The expression levels of TGFß, PDGFRα, USP-28, Ki67, and PDGFA were decreased significantly in MC applied groups. Renal TAS levels were significantly lower in group-3. Liver and kidney OSI values were increased in both groups 2 and 3. CONCLUSION: The results indicated that low dose 3-MC caused oxidative stress and downregulated apoptotic and cytokine markers in the long term and cisplatin had no ameliorative effects on this degeneration processes (Tab. 3, Fig. 3, Ref. 32). Text in PDF www.elis.sk.

Novel amoxicillin nanoparticles formulated as sustained release delivery system for poultry use.

Amoxicillin is used in the treatment and prevention of a wide range of diseases in poultry breeding. However, its short half-life and low bioavailability restrict its clinical application in these species. Entrapment of drugs into polymeric nanoparticles (nps) presents a means to improve gastrointestinal absorption and oral bioavailability of drugs. This study was aimed to overcome limitation of amoxicillin use in poultry breeding. Amoxicillin was loaded into sodium alginate-polyvinyl alcohol (NaAlg-PVA) blend nps, and characterization of the prepared nps was performed. For pharmacokinetic study, commercial male broilers were used and comparative pharmacokinetics of free and nanoparticle form of amoxicillin were investigated. Twenty-one broilers were divided into three groups. All groups received 10 mg/kg drug. Blood samples were collected, and drug plasma concentrations were determined by HPLC. The results demonstrated that the particle size, zeta potential, encapsulation efficiency, and loading capacity of the nps were 513.96 ± 19.46 nm, -45.36 ± 1.35 mV, 43.66 ± 3.30, and 12.06 ± 0.83%, respectively. In vitro drug release exhibited a biphasic pattern with an initial burst release of 18% within 2 hr followed by a sustained release over 22 hr. The pharmacokinetic results showed that amoxicillin nps have higher bioavailability and longer plasma half-life (p < .01) than free amoxicillin. These results indicate that amoxicillin nano formulation is suitable for oral administration in broilers.

Measurement of C-reactive protein and prostaglandin F2α metabolite concentrations in differentiation of canine pyometra and cystic endometrial hyperplasia/mucometra.

Canine pyometra is a dioestrus period disease in which systemic inflammatory response syndrome (SIRS) is a common outcome due to the response of the body to the bacterial infection. The purpose of this study was i) to differentiate canine pyometra and cystic endometrial hyperplasia (CEH)/mucometra by measuring serum C-reactive protein (CRP) and prostaglandin F2α metabolite (PGFM) concentrations in blood and ii) to compare serum concentrations of CRP and PGFM in bitches with a pathological uterus (pyometra or CEH/mucometra) to concentrations in bitches with a healthy uterus. Mean CRP concentrations were found significantly higher (p < 0.001) in dogs with pyometra compared to those with CEH/mucometra or healthy uterus. However, no statistical difference could be detected between the groups for mean PGFM concentrations. Mean white blood cell count (WBC), alkaline phosphatase (ALP) and total protein concentrations were found significantly higher (p < 0.001) in dogs with pyometra. Escherichia coli was the most frequently isolated microorganism from dogs with pyometra (64.3%). Edwardsiella spp. was detected in a single case of pyometra for the first time. In conclusion, our results demonstrate that serum CRP concentrations were increased in dogs with pyometra and thus we conclude that serum CRP concentration but not PGFM might be useful as a marker to differentiate a case of CEH/mucometra from pyometra in female dogs. To the authors' knowledge, this is the first report in which Edwardsiella spp. has been isolated in the canine uterus.

Endometrial adenocarcinoma in two young queens.

Endometrial adenocarcinoma is described in two young intact female Persian cats presented with haemorrhagic vaginal discharge. The discharge had been present for 3 and 15 days in each case, respectively. On the basis of signalment, history, clinical, laboratory and diagnostic imaging findings a tentative diagnosis of pyometra was made in each cat and ovariohysterectomy was subsequently performed. Histological examination revealed endometrial adenocarcinoma in both cats together with ovarian luteal cysts and pyometra in one case. Immunohistochemistry revealed a diffuse and strongly positive reaction for oestrogen receptors in the nuclei of glandular epithelium in well-differentiated areas of the neoplastic tissue. Poorly differentiated areas of solid growth pattern had weak and focal expression of oestrogen receptors. Furthermore, the neoplastic cells within these areas displayed moderate to strong diffuse nuclear staining with Ki-67 antibody suggestive of a high proliferative capacity. The owners reported that the cats were clinically healthy 23 and 21 months after the surgery. These cases highlight the importance of histological examination of surgically resected tissue even when clinical signs and laboratory findings suggest pyometra.

Determination of melamine in milk and dairy products by high performance liquid chromatography.

A simple, precise, accurate, and validated reverse-phase HPLC method was developed for the determination of melamine in milk (pasteurized and UHT milk) and dairy products (powdered infant formula, fruit yogurt, soft cheese, and milk powder). Following extraction with acetonitrile:water (50:50, vol/vol), samples were purified by filter (0.45 µm), separated on a Nucleosil C8 column (4.6 mm × 250 mm, 3 µm) with acetonitrile:10 mmol/L sodium L-octane sulfonate (pH 3.1; 15:85, vol/vol) as mobile phase at a flow rate of 1 mL/min, and determined by a photodiode array detector. A linear calibration curve was obtained in the concentration range from 0.05 to 5 mg/kg. Milk and dairy products were fortified with melamine at 4 levels producing average recovery yields of 95 to 109%. The limits of detection and quantification of melamine were 35 to 110 and 105 to 340 µg/kg, respectively. The method was then used to analyze 300 samples of milk and dairy products purchased from major retailers in Turkey. Melamine was not found in infant formulas and pasteurized UHT milk, whereas 2% of cheese, 8% of milk powder, and 44% of yogurt samples contained melamine at the 121, 694±146, and 294±98 µg/kg levels, respectively. These findings were below the limits set by the Codex Alimentarius Commission and European Union legislation. This is the first study to confirm the existence of melamine in milk and dairy products in Turkey. Consumption of foods containing these low levels of melamine does not constitute a health risk for consumers.

Fine-needle aspiration biopsy of canine mammary gland tumours: a comparison between cytology and histopathology.

In the current study, a total of 90 mammary neoplasms obtained from 55 female dogs were used to determine the accuracy of fine-needle aspiration cytology in the diagnosis of canine mammary tumours and to investigate the feasibility of this technique for the differentiation of simple tumours from complex or mixed tumours. Three aspirations were performed on each mammary gland mass using a 22-gauge needle attached to a 5-ml syringe before the mammary glands were surgically excised and submitted for histopathological examination. Twenty-five (27.7%) of 90 samples were classified as insufficient/inadequate for diagnosis. Of the remaining 65 samples, six (9.2%) were benign, 51 (78.5%) were malignant tumours and 8 (12.3%) were suspicious. Histopathological examination of the 90 specimens revealed five (5.6%) benign, 84 (93.3%) malignant and one (1.1%) non-neoplastic lesion. The diagnostic accuracy, sensitivity and specificity of cytologic examination for diagnosing malignancy were 96.5%, 96.2% and 100%, respectively. However, when inadequate (n = 25) and suspicious (n = 8) samples were included, the diagnostic accuracy and sensitivity decreased to 63.3% and 60.7%, respectively, but no change was observed in the specificity. Furthermore, it was not possible to differentiate simple tumours from complex and mixed tumours because spindle cells were seen in both 28% of the simple tumours and 39.3% of the complex or mix tumours. In conclusion, we believe that fine-needle aspiration cytology of canine mammary tumours is a valuable diagnostic tool, although our results indicated lower accuracy when inadequate samples were taken into consideration.

Vaginal neurofibroma in a hysterectomized poodle dog.

A 15-year-old, spayed, female poodle dog was presented for evaluation of a mass of tissue prolapsed from the vulva. The dog had been hysterectomized when it was 5 years old. A vaginal mass had been removed approximately 10 months before presentation. Haematological and serum biochemistry analyses demonstrated mild leucocytosis and glycaemia. A vaginal smear was predominantly made up of parabasal cells and intermediate cells with no neoplastic cells. Thoracal and abdominal radiographic findings were unremarkable. The ovaries could not be identified using abdominal ultrasonography. A midline exploratory laparotomy identified both ovaries that were surgically excised. The vaginal mass was also removed following an episiotomy procedure. Histopathological examination of the mass demonstrated that it was a neurofibroma. Both ovaries had cystic changes. Four months after the surgery, the owner reported that the dog was clinically normal. To the authors' knowledge, this is the first reported case of a vaginal neurofibroma after an incomplete ovariohysterectomy in the dog.

Uterine leiomyoma in a spayed poodle bitch: a case report.

A 16-year-old, spayed, female poodle dog was presented for evaluation of tenesmus, with the ability to attract male dogs and a willingness to mate. The dog had undergone an ovariohysterectomy when it was 7 years old. Haematological and serum biochemistry analyses demonstrated leucocytosis and a mild uraemia. A vaginal smear was predominantly made up of superficial cells, accounting for at least 80% of the cells. Abdominal ultrasonography revealed a heterogeneous multicystic mass at the caudal abdomen and two masses with anechoic areas adjacent to the caudal pole of each kidney. A midline exploratory laparotomy identified a uterine mass and residual ovaries that were surgically excised. Histopathological examination of the mass demonstrated that it was a leiomyoma. The left ovary had a cystic structure and the right ovary had a papillary cystic adenoma. Seven months after the surgery, the owner reported that the dog was clinically normal. To the authors' knowledge, this is the first reported case of a uterine leiomyoma after an incomplete ovariohysterectomy in the dog.

Difference in drug resistance patterns between minor HIV-1 populations in cerebrospinal fluid and plasma.

OBJECTIVE: The aim of the study was to determine to what extent unique drug resistance patterns appear in minor and major HIV-1 quasispecies in cerebrospinal fluid (CSF) as compared with blood. METHODS: Forty-four plasma and CSF samples from 13 multi-treatment-experienced patients, seven of whom provided longitudinal samples, were included in the study. The subjects had failed antiretroviral therapy including lamivudine. The reverse transcriptase (RT) gene was examined by selective real-time polymerase chain reaction (SPCR), which can detect M184I/V mutants down to 0.2% of the viral population. RESULTS: SPCR revealed differences at amino acid position 184 in the plasma/CSF populations in 12 paired samples from eight patients. One plasma sample was positive by SPCR where direct sequencing showed wild-type M184. The other 11 paired samples showed quantitative differences in the mixed populations of the mutant or wild-type M184 quasispecies. Differences in other resistance-associated mutations between plasma and CSF viruses were also found by direct sequencing. CONCLUSIONS: In multi-treatment-experienced patients with therapy failure, differences in drug resistance patterns were found frequently between plasma and CSF in both minor and major viral populations. To what extent this was a true biological phenomenon remains to be established, and the clinical relevance of these findings is yet to be determined.

Perineal hernia because of retroflexion of the urinary bladder in a rottweiler bitch during pregnancy.

A young pregnant rottweiler bitch was presented with a large mass of tissue protruding through the vulvar lips. Foetal death was diagnosed on ultrasonography, and ovariohysterectomy was performed, at which time the bladder was observed to be dislocated caudally into the pelvic cavity under the vagina. The bladder was manually repositioned during surgery. One week later, the bitch was healthy with no evidence of dysuria, stranguria or urinary incontinence. Two months later, the owner reported that the bitch was clinically normal with no recurrence of the retroflexion. To the authors' knowledge, this is the first reported case of retroflexion of the urinary bladder during pregnancy.

Short-term effects of prepubertal ovariohysterectomy on skeletal, physical and behavioural development of dogs up to 24 weeks of age.

This study investigated the effects of ovariohysterectomy performed at 10 weeks of age on body weight, behaviour, bone length, serum calcium and phosphorus concentrations and gonadal hormone (progesterone and oestradiol) levels until 24 weeks of age in 31 clinically healthy mixed-breed female puppies. The littermates were randomly assigned to three groups. Group I (n=11) was completely ovariohysterectomised, Group II (n=10) was incompletely ovariohysterectomised with one ovary left intact, and Group III (n=10) was sham operated. No significant differences were found between the groups for any of the parameters evaluated. It was determined that, except for two measurements of two animals in Group II at 20 and 24 weeks of age, all animals had undetectable concentrations of oestradiol 17-beta. These findings suggest that an ovariohysterectomy performed on a mixed-breed dog at 10 weeks of age does not affect her skeletal, behavioural or physical development in the period studied, i.e. until 24 weeks of age.

The effect of prepubertal ovariohysterectomy on spine 1 mineral density and mineral content in puppies: a preliminary study.

This pilot study investigated early effects of prepubertal ovariohysterectomy on bone mineral density (BMD) and bone mineral content (BMC) of lumbar vertebraes (L2-L5 and L Total) and on serum biochemistry. Both prepubertal ovariohysterectomy (Group Po: n=3) and sham surgery (Group Lp: n=3) were performed at 10 weeks of age. Blood samplings, BMD and BMC measurements of L2, L3, L4, L5 and L Total were performed at 12, 16, 20 and 24 weeks of age. No significant difference (P>0.05) in BMD and BMC was observed between the groups during the experimental period. The difference in serum calcium and phosphorus concentrations was found to be statistically unsignificant (P>0.05). These results fail to demonstrate a difference in BMD and BMC of lumbar vertebraes (L2-L5) as well as serum calcium and phosphorus levels after prepubertal ovariohysterectomy until six months of age in mixed breed puppies, but they should be considered only descriptive because of the very low sample number and of the low power of the statistical analyses.

Effect of prepubertal ovariohysterectomy on bone mineral density and bone mineral content in puppies.

In this study, dual energy X-ray absorptiometry (DXA) was performed on the calcaneus (CAL) and accessory carpal bone (ACB) of early-age neutered (n = 11) and sham-operated puppies (n = 10) to investigate changes in bone mineral density (BMD) and bone mineral content (BMC) over time. Prepubertal ovariohysterectomy and sham surgery were performed at 10 weeks, while BMD and BMC were measured at 12, 16, 20 and 24 weeks of age. BMD and BMC of CAL and ACB of ovariohysterectomised puppies were found to be higher. The differences between the groups were statistically unimportant; however, the fourth measurement of the ACB showed a significant (P < 0.05) difference. Sexually intact puppies weighed less than gonadectomised puppies in all measurements. Because of the positive correlations between body weight and bone mineral density, BMD and BMC of early-age neutered puppies were found to be higher. These results suggest that BMD and BMC are not affected by prepubertal ovariohysterectomy until six months of age in mixed-breed puppies.

Use of aglepristone and aglepristone + intrauterine antibiotic for the treatment of pyometra in bitches.

In this study, the efficacy of aglepristone and/or intrauterine antibiotic administration for the treatment of bitches with cystic endometrial hyperplasia/pyometra complex was investigated. Twenty-four bitches (5-12 years old) with the diagnosis of pyometra were treated at the University of Kafkas and at Istanbul University. The diagnosis of pyometra was established on the basis of the results of clinical, ultrasonographic and vaginal examinations, the haematological and biochemical findings and the history data. In Group I (n = 13), aglepristone (Antiprogestin, Alizine, Virbac, France; 0.33 ml/kg, s.c.) was administered on days 1, 2, 7, and 14 (day 1: diagnosis). In Group II (n = 11), intrauterine antibiotic treatment was performed according to the antibiogram on days 1, 2, 4, 6, 8 in addition to aglepristone given as in Group I. Clinical and ultrasonographic examinations, haematological results and occurrence of oestrous cycles revealed that the ratio of effectively treated bitches was 6/13 and 9/11 in Groups I and II, respectively.