Different expression of Defensin-B gene in the endometrium of mares of different age during the breeding season.

BACKGROUND: Despite being one of the major causes of infertility in mares, the mechanisms responsible for equine endometrosis are still unclear and controversial. In the last few years, many investigations focused on local immune response modulation. Since it is generally accepted that endometrial fibrosis increases with age, we hypothesize that older mares could show altered local immune modulation, initiating a pro-inflammatory and tissue remodeling cascade of events that could lead to endometrosis. The aim of this study, indeed, is to evaluate and describe the local gene expression of genes involved in acute inflammatory response and fibrosis (COL1A1, COL3A1, TNFA, MMP9, IL6, TGFB1 and TGFBR1), together with others associated to immune modulation (DEFB4B, IDO1 and FOXP3), in uterine specimens from mares of different age. RESULTS: Twenty-five Standardbred mares were involved in the study with age ranging from 7 to 19 years (mean 10.40 ± 4.42). They were divided by age into two groups: G1 (n = 15, less than 10 years old) and G2 (N = 10, greater than 11 years old). Specimens from the uterus' right horn-body junction were collected and processed for histology evaluation and RT-qPCR assay.Gene expression of DEFB4B, MMP9 and TNFA was higher in younger mares, suggesting a balance in immune modulation and tissue remodeling. Interleukin-6 and COL3A1 gene expressions were greater in older animals, probably indicating inflammatory pathways activation and fibrosis increase. Although no differences in fibrosis and inflammation distribution could be found with histological examination among G1 and G2, our results suggest a possible involvement of DEF4BB in regulating the local immune response in younger mare's uterus (G1); age may contribute to the dis-regulation of DEFB4B transcription and, indirectly, influence the extracellular matrix homeostasis. Transcription of IDO1 and FOXP3 genes, instead, does not seem to be age related, or to be involved in local immune-response and tissue remodeling functions. CONCLUSIONS: Further investigations are needed in order to clarify the interactions between the expression of DEFB4B, IL6, TNFA, COL3A1 and MMP9 and other local signals of immune-modulation and tissue remodeling, in mares in a prospective study design.

E-cadherin, N-cadherin Expression and Histologic Characterization of Canine Choroid Plexus Tumors.

Choroid plexus tumors (CPTs) are reported with an increasing incidence in dogs, and they call for a reexamination of histologic features and criteria of classification corresponding to their biological behavior. In this study, the human World Health Organization classification was applied to 16 canine CPTs, and the expression of molecules involved in neoplastic cell adhesion (E-cadherin, N-cadherin), invasion (doublecortin), and proliferation (Ki-67) was investigated. Mitotic index was found to be the main criterion for grading CPTs. Cell density and multilayering of papillae were also statistically associated with histologic grade. Intraventricular spread and parenchymal invasion was observed for tumors showing histologic benign features. E-cadherin was expressed in all CPT grades, independent of tumor invasion. N-cadherin immunolabeling was more expressed in grade I than high-grade CPTs, whereas doublecortin expression was not detected in CPTs. An increasing proliferative activity was observed in relation with histologic grade.

Case Report: Adjuvant image-guided radiation therapy reduces surgical invasiveness in malignant peripheral nerve sheath tumors.

Introduction: Malignant peripheral nerve sheath tumors (MPNSTs) are a group of rare soft tissue sarcomas of mesenchymal origin. These tumors generally require extensive local excision owing to their aggressive potential. Though the role of radiotherapy is controversial, in this report, we present the case of an MPNST in the forearm that was treated with microsurgery followed by image-guided radiation therapy to achieve complete tumor disappearance at the 18-month follow-up. Case report: A 69-year-old woman with underlying paranoid schizophrenia was referred to our department with pain, severe swelling, and ecchymosis of her right forearm. Physical examination showed hypoesthesia in the segments innervated by the median nerve and reduced motor strength of her right hand. A gadolinium-enhanced MRI showed a large malignant peripheral nerve sheath tumor (13 x 8 x 7 cm) of the median nerve in the forearm. She underwent microsurgical en-bloc tumor resection with sparing of the median nerve. Thirty-five days postoperatively, she underwent image-guided radiotherapy (IGRT) using volumetric modulated arc therapy (VMAT). Serial MRI scans of the forearm with Gadolinium and whole-body CT scan with contrast enhancement at 30 days, 6 months, 1 year, and 18 months postoperatively documented no tumor recurrence, remnants, or metastases. Conclusions: In this report, we demonstrate the successful use of advanced radiotherapy techniques such as IGRT while avoiding demolitive surgery for MPNST. Though a longer follow-up is necessary, at the 18-month follow-up, the patient demonstrated good outcomes from surgical resection followed by adjuvant RT for MPNST in the forearm.

Comparison of Staged Stereotactic Radiosurgery and Fractionated Stereotactic Radiotherapy in Patients with Brain Metastases > 2 cm without Prior Whole Brain Radiotherapy: A Systematic Review and Meta-Analysis.

OBJECTIVE: To compare fractionated stereotactic radiotherapy (FSRT) with staged stereotactic radiosurgery (SSRS) in patients with brain metastases >2 cm without prior whole brain radiotherapy. METHODS: In this systematic review and meta-analysis, PubMed, Scopus, Web of Science, Embase, and Cochrane were searched to include studies that evaluated FSRT and/or SSRS for brain metastases >2 cm or 4 cm3 in adult patients with a known primary malignancy and no prior history of whole brain radiotherapy. Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines were followed and an indirect random-effect meta-analyses was conducted to compare treatment outcomes between the two modalities. RESULTS: A total of 10 studies were included, comprising 612 (778 metastases) and 250 patients (265 metastases) in the SSRS and FSRT groups, respectively. The SSRS group had significantly older patients (66.6 ± 17.51 years vs. 62.37 ± 37.89 years; P = 0.029) with lower rate of control of primary disease (11.59% vs. 78.7%, P < 0.00001), and more patients with Karnofsky performance status ≥70 at baseline (92.81% vs. 88.56%; P = 0.045). FSRT was associated with a statistically nonsignificant but clinically important lower 12-month overall survival (44.75% [95% confidence interval [CI]: 30.48%-59.95%] vs. 53.25% [95%CI: 45.15%-61.19%], P = 0.1615) and higher rate of salvage radiotherapy (18.18% [95%CI: 8.75%-34%] vs. 12.27% [95%CI: 5.98%-23.53%], P = 0.0841). Both groups had comparable rates of local tumor control, mortality, tumor progression, recurrence, neurological death, and 6-month overall survival. CONCLUSIONS: SSRS and FSRT were found to be comparable for treating brain metastases >2 cm not previously irradiated. Given the paucity of such studies, trials directly comparing the two treatment strategies are warranted to support these findings.

A morphological and quantitative immunohistochemical study of the interstitial cells of Cajal in the normal equine intestinal tracts.

REASONS FOR PERFORMING STUDY: The interstitial cells of Cajal (ICC) play a key role in the control of intestinal motility and have been implicated in several human gastrointestinal dysmotility syndromes, in equine grass sickness and in other intestinal disorders where a significant reduction in ICC density was observed. OBJECTIVES: To investigate the density of ICC in clinically normal horses, ICC c-Kit expression was evaluated by image analysis in order to obtain numerical data. METHODS: Intestinal samples from the jejunum to small colon from 5 clinically normal horses were studied. Immunohistochemical labelling of ICC was performed using an anti-c-Kit antibody. Density of ICC was calculated using image analysis software. RESULTS: In the equine intestinal tract 2 types of ICC were observed: intramuscular ICC, i.e. ICC in the internal circular layer (IC-CM) and ICC in the external longitudinal layer (IC-LM), and myenteric ICC (IC-MY). The density of IC-MY was found to be higher throughout the small intestine. IC-MY density in the large intestine appeared to be greatest in the right ventral colon and in the small colon. IC-MY density in the ileocaecal junction showed an intermediate value compared to the small and large intestine. On the other hand, the density of IC-CM was found to be higher in the ileocaecal junction, whereas the caecum, left ventral colon and the left dorsal colon showed the lowest c-Kit immunoreactivity. The ileal tract and the ileocaecal junction showed an appreciable IC-LM density. CONCLUSIONS: Image analysis is a rapid and reproducible method to establish the density of ICC in the normal equine intestinal tract. POTENTIAL RELEVANCE: This study corroborates the findings of previous studies and provides a platform for further future pathological investigations of the equine intestine by supplying usable numerical data as comparative elements.

Intradural extramedullary granular cell tumour in a cat.

A 7-year-old domestic shorthair cat was evaluated for progressive paraparesis, inability to jump, a paralysed tail and inability to void the bladder. Neurologic examination was consistent with a L4-S3 localisation. Survey radiographs of the lumbar vertebral column revealed L4-L7 vertebral body remodelling. A pre-contrast T1-weighted hyperintense, diffusely enhancing intradural lesion extending from L4 to S1 vertebral bodies was detected by MRI. Large, mesenchymal, round-to-polygonal cells arranged in nests or sheets were found on histologic examination at post mortem. These cells were characterised by abundant intracytoplasmic PAS-positive, diastase-resistant granules and positive immunoexpression of vimentin, S-100, neuron-specific enolase and desmin. This is the first report of a spinal granular cell tumour in a cat.

Matrix metalloproteinase-2 and matrix metalloproteinase-9 expression in canine and feline meningioma.

Fifty-one meningiomas obtained from 28 dogs and 23 cats were selected for this study to investigate the immunohistochemical expression of matrix metalloproteinase (MMP)-2 and MMP-9 and to compare it to the reverse transcriptase subunit of human-telomerase, progesterone receptor expression, and the proliferative index of the tumors, expressed by Ki67 and proliferating cellular nuclear antigen. Paraffin-embedded tumor tissue was obtained from biopsy samples (28 cases) and at necropsy (23 cases). The most common histotype was malignant in dogs (12/28) and transitional in cats (12/23). Slides immunolabelled for MMPs showed a diffuse cytoplasmic pattern. Twenty-one cases (19 dogs and 2 cats) did not express MMP-2, while only 2 cases were completely negative for MMP-9. The highest values of MMP-2 and MMP-9 were observed in a psammomatous and meningothelial tumor, respectively. On statistical analysis, MMP-2 expression did not show a significant correlation with MMP-9. Moreover, both MMP expressions failed to show significant variance among histologic patterns of the tumor and correlation with the proliferative index. MMP immunolabeling showed an inconstant correlation with progesterone receptor expression. No significant correlation was found between MMP and reverse transcriptase subunit of human-telomerase expression. In feline meningiomas, the MMP-2 value was significantly higher than in canine tumors and the MMP-9 value tended to be low for meningiomas with a follow-up duration from the 23(rd) month to the 44(th) month. In cats, the longer the time from surgery, the lower the proliferative index seemed to be. In dogs, we failed to find a correlation between MMP expression and the follow-up duration.

Somatostatin Receptor 2 Expression in Canine Meningioma.

The neuropeptide somatostatin (SST) plays an important regulatory role in the proliferation of normal and neoplastic cells. Five subtypes of somatostatin receptors (SSTRs), SSTR1-SSTR5, have been identified in human tumours. The SSTR2 subtype is identified most commonly in meningiomas. Long half-life SST analogues are now recommended for the systemic treatment of unresectable or radiation-refractory recurrent human meningiomas. In this study, SSTR2 expression was evaluated in 46 canine meningiomas; in 21 cases this was by immunohistochemistry and in 25 cases by reverse transcriptase quantitative polymerase chain reaction (RT-qPCR). In addition, SSTR2 expression was evaluated by immunocytochemistry, western blotting and RT-qPCR on primary cell cultures prepared from two canine meningiomas. SSTR2 immunohistochemical expression was observed in 17/21 cases (81%), and SSTR2 mRNA expression was detected in 14/25 cases (56%). SSTR2 protein and gene expression were not significantly correlated with the tumour histological subtype or grade. Overall, meningothelial meningiomas showed constant and diffuse SSTR2 immunohistochemical expression and the highest SSTR2 gene expression level compared with other subtypes. A tendency for loss of SSTR2 in high-grade meningiomas was observed in both immunohistochemical and RT-qPCR studies. About 90% of cultured canine meningioma cells showed SSTR2 expression. In both of the meningioma cell cultures, SSTR2 expression was also detected by western blotting and RT-qPCR. This study demonstrates for the first time that canine meningioma expresses SSTR2 and that this expression is maintained in vitro. Our results, while preliminary, provide encouragement for further studies aimed at finding novel medical treatment strategies for canine meningioma, especially for tumours that are not surgically accessible.

Porencephaly and Periventricular Encephalitis in a 4-month-old Puppy: Detection of Canine Parvovirus Type 2 and Potential Role in Brain Lesions.

A 4-month-old puppy died after showing intracranial signs a few days after a suspected viral enteritis. Grossly, the right cerebral hemisphere had a large irregular cavity external to the internal capsule. Histopathological examination revealed a cystic lesion in the right hemisphere and non-suppurative inflammation of the diencephalon and periaqueductal nervous tissue. Porencephaly associated with periventricular non-suppurative encephalitis was diagnosed. A nested polymerase chain reaction (PCR) identified the presence of parvovirus DNA in the brain and real-time PCR typed this as canine parvovirus (CPV) type 2a. Immunohistochemistry revealed the presence of CPV antigen in the cytoplasm of scattered cells in the subependymal layers and choroid plexus epithelium. The porencephaly was not associated with inflammatory lesions or CPV antigen and was considered to have preceded the neurological signs. In contrast, the detection of CPV antigen in the subependymal layers and choroid plexus epithelium supported the association of this virus with the periventricular encephalitis.

Biology, prognosis and response to therapy of breast carcinomas according to HER2 score.

BACKGROUND: The standardization of the HER2 score and recent changes in therapeutic modalities points to the need for a reevaluation of the role of HER2 in recently diagnosed breast carcinoma. PATIENTS AND METHODS: A multicenter, retrospective study of 1794 primary breast carcinomas diagnosed in Italy in 2000/2001 and scored in HER2 four categories according to immunohistochemistry was conducted. RESULTS: Ductal histotype, vascular invasion, grade, MIB1 positivity, estrogen and progesterone receptor expression differed significantly in HER2 3+ tumors compared with the other categories. HER2 2+ tumors almost showed values intermediate between those of the negative and the 3+ subgroups. The characteristics of HER2 1+ tumors were found to be in between those of HER2 0 and 2+ tumors. With a median follow-up of 54 months, HER2 3+ status was associated with higher relapse rates in node-positive and node-negative subgroups, while HER2 2+ only in node positive. Analysis of relapses according to type of therapy provided evidence of responsiveness of HER2-positive tumors to chemotherapy, especially taxanes. CONCLUSIONS: The present prognostic significance of HER2 is correlated to receptor expression level and points to the need to consider HER2 2+ and HER2 3+ tumors as distinct diseases with different outcomes and specific features.

Toxoplasma gondii brain granuloma in a cat: diagnosis using cytology from an intraoperative sample and sequential magnetic resonance imaging.

A cat with a history of seizures and clinical suspicion of forebrain disorder underwent a brain magnetic resonance imaging. A space-occupying lesion was identified in the left temporal lobe. The mass was surgically removed, and cytological, histological and immunohistochemical examinations documented the presence of Toxoplasma gondii. A definitive diagnosis of an intracranial T gondii granuloma was made. The cat was treated with clindamycin and phenobarbital and the seizures did not recur. After 10 months, a second magnetic resonance imaging showed severe brain atrophy, but T gondii granuloma recurrence was not noted. Twenty-one months after surgery, the cat's condition deteriorated, and another magnetic resonance imaging showed a presumptive recurrence of T gondii granuloma. In cats, T gondii granuloma must be considered as a differential diagnosis even when only a single intracranial mass is present. Cytology and magnetic resonance imaging can be useful in making a definitive diagnosis and to follow the evolution of the lesion.

Sympathetic Innervation and Adrenergic Receptors in Equine Deep Digital Flexor Tendinopathy: Preliminary Results.

The aim of this study was to delineate the pattern of sympathetic innervation in the suprasesamoidean region of the deep digital flexor tendon (DDFT) in horses with tendinopathy by immunohistochemical labelling for tyrosine hydroxylase (TH) and α-1 adrenergic receptor (α1-AR). Twelve forelimbs were obtained from 10 horses with DDFT tendinopathy and six feet obtained from six horses were used as healthy controls. Post-mortem radiographic, ultrasonographic and gross examinations were performed on the suprasesamoidean area of the DDFT to assess the presence of tendinopathy. Longitudinal sections were collected and processed. Lesions were classified as core lesions, dorsal border lesions and parasagittal oblique splits. Immunohistochemistry was performed and the degree of immunoreaction was classified as absent, mild or marked. Seven core lesions, four dorsal border lesions and one parasagittal oblique split were identified. There was no increased expression of sympathetic innervation in samples with a dorsal border lesion of the DDFT compared with healthy samples. In contrast, core lesions showed increased expression of α1-AR and reduced expression of TH, which supports the hypothesis of a compensatory imbalance between the sympathetic mediator and the sympathetic receptors as a cause or effect of structural damage. In addition, adrenergic activation could stimulate cell proliferation and differentiation within these lesions.

Immunohistochemical expression of cyclooxygenase-2 (COX-2) is not associated with tumor grade in feline meningiomas.

Meningioma is the most common primary brain tumor in cats and occurs less frequently in the spinal cord. This study aimed to investigate cyclooxygenase-2 (COX-2) expression in feline meningiomas, and the possible association between COX-2 immunoreactivity and tumor grade using eight low-grade and seven high-grade meningiomas. All tumors (n=15/15) were immunoreactive to COX-2. The expression of COX-2 was not significantly correlated with tumor grade (P=0.22 and 0.34 for staining and intensity, respectively) but was significantly associated with necrosis (P=0.04 and 0.01 for staining and intensity, respectively). The findings in this study suggest that feline meningiomas express COX-2, but there were no differences in COX-2 immunoreactivity patterns between low- and high-grade meningiomas. However, the association between COX-2 expression and the presence of necrosis indicates a potential area for therapeutic intervention with selective COX-2 inhibitors.

Cerebellar leptomeningeal carcinomatosis in a dog.

Diffuse cerebellar meningeal carcinomatosis secondary to haematogenous dissemination from an anaplastic solid mammary carcinoma was diagnosed in an old German shepherd dog suffering from seizures and rapidly progressing to stupor. A single computed tomography cerebellar scan identified an unusual homogeneous density area that was considered to be associated with a vascular disorder, in the absence of space-occupying lesions. At necropsy, nodular masses were observed in the mammary gland, lungs, tracheobronchial lymph nodes and adrenals. Cerebellar leptomeninges were affected by diffuse blood effusion. Histology showed a solid mammary tumour, characterised by anaplastic cells with a cytoplasmic keratin-positive and vimentin-negative immunoreaction. The tumour had spread to the lungs, tracheobronchial lymph nodes and adrenals. Cerebellar leptomeninges were diffusely infiltrated by the cytokeratin-positive neoplastic cells. Even though computed tomography scan gave no evidence of meningeal carcinomatosis, it was considered that a cerebellar vascular disorder might be present. This was subsequently confirmed by neuropathological investigation and seen to be associated with a cerebellar leptomeningeal carcinomatosis.

Gene Expression of Matrix Metalloproteinases and their Inhibitors (TIMPs) in Meningiomas of Dogs.

BACKGROUND: Matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) are considered to be key mediators of tumor invasion and metastasis. MMP-2 and MMP-9 are expressed in meningiomas of dogs, but TIMP expression, and variations of specific MMP/TIMP ratios still are unknown in this tumor. HYPOTHESIS/OBJECTIVES: Expression of MMP/TIMP might increase progressively from grade I to grade III meningioma. Therefore, genetic expression of MMP-2 and MMP-9, and specific TIMP-2 and TIMP-1, respectively, has been investigated in meningiomas of different grades. ANIMALS: Selected formalin-fixed paraffin-embedded tissue from 43 meningiomas of dogs was evaluated. METHODS: Genetic material was obtained from pathologic samples and used for quantitative reverse transcriptase real-time polymerase chain reaction (RT-qPCR). RESULTS: MMP-9 was not expressed in all of the tumors, but MMP-2 was significantly more expressed in papillary meningioma. Likewise, the MMP-2/TIMP-2 ratio was numerically higher in papillary meningiomas compared to all grades (>3.5 times) showing a strong bias in favor of metalloproteinase. In the papillary meningioma, TIMP-1 gene expression was significantly higher than in grades I and III. CONCLUSIONS AND CLINICAL IMPORTANCE: MMP-2/TIMP-2 imbalance might contribute to the aggressive biologic behavior of papillary meningiomas in dogs. TIMP-1 expression may play a role independent of MMP-9 expression in neoplastic progression. These results further support that therapeutic and prognostic evaluations of dogs with meningioma need to be addressed according to different histologic patterns as is performed in humans.

Sympathetic innervation of the suprasesamoidean region of the deep digital flexor tendon in the forelimbs of horses.

The purpose of this study was to delineate the pattern of sympathetic innervation in the suprasesamoidean region of the deep digital flexor tendon (DDFT) in horses using immunohistochemical staining (IHC) for tyrosine hydroxylase (TH) and alpha-1 adrenergic receptor (α1-AR). Fourteen forelimbs were collected from 10 horses. Longitudinal sections of the suprasesamoidean region of healthy DDFTs were harvested. Most of the sympathetic innervation was found to be in the walls of blood vessels. The tendon tissue proper was sparsely innervated, with a lesser degree of innervation within the dorsal fibrocartilage. Increased α1-AR immunostaining was also detected in walls of blood vessels and in spindle cells of fibrocartilage. Both α1-AR and TH immunostaining were detected in tenocytes. These findings support the presence of autocrine/paracrine catecholaminergic signalling in equine tendon tissue.

The force of measles infection in East Africa.

Catalytic models were applied to age-specific incidence data from two East African studies in order to study the force of measles infection in relation to age. The results are compared with the pattern observed in England and Wales. In East Africa the force of measles infection appears to be independent of age, probably as a result of sociocultural factors. Consequently the incidence of measles is highest in the second half of the first year of life and decreases with increasing age. Major vaccination efforts will raise the average age of measles attack, but the peak incidence will remain in the first year of life. This complicates the decision on the optimal age for vaccination and the need for incidence surveillance is stressed. The simple catalytic model, which is based on the assumption of an age-independent force of infection may play an important role in measles surveillance. With this model simple and economic surveillance studies could be designed which use cross-sectional data only.

Neosporosis in a dog in Italy.

Clinical neosporosis was diagnosed in a 2-month-old Pit Bull Terrier from Italy. Neospora caninum tachyzoites were found in semitendinosus muscle with myositis. The diagnosis was confirmed by immunohistochemical staining with anti-N. caninum specific antibodies. This is the first report of systemic neosporosis in dogs from Italy.

Immunohistochemical identification and image analysis quantification of oestrogen and progesterone receptors in canine and feline meningioma.

Oestrogen and progesterone receptor expression has been detected in human meningioma; in canine and feline meningiomas, however, the identification of steroid receptors requires further investigation. Fourteen meningiomas obtained from nine dogs and five cats were examined immunohistochemically for oestrogen receptors (ORs) and progesterone receptors (PRs). The immunolabelling reaction was quantified by light microscopy and image analysis. ORs were expressed in low numbers in two feline meningiomas and in one canine meningioma. PRs were more numerous, with more extensive tissue labelling. Conventional linear regression analysis showed that OR expression was not significant as compared with PR expression. PR expression was lowest in meningiomas with a high proliferation index, as determined by Ki67 expression. Conventional linear regression analysis between PR and Ki67 concentration confirmed a significant indirect relationship.

Female genital mutilation in Nigeria.

OBJECTIVES: To determine the prevalence and distribution of female genital mutilation (FGM) procedures in a Nigerian population. METHODS: Five hundred consecutive women were evaluated for evidence of FGM. The WHO classification system was utilized. Demographic and sociocultural information was collected. RESULTS: Thirty-four percent of women were found to have some type of FGM. Type I and Type II procedures were the most common. Fifty-five percent of women were unaware they had FGM and 62% with FGM did not know the reason. Twenty-one percent of women said they were going to have FGM on their daughters. CONCLUSIONS: FGM is still broadly practiced among ethnic and religious groups in Nigeria. Educational efforts need to be directed at eliminating this practice.