249 research outputs found
Molecular-based tumour subtypes of canine mammary carcinomas assessed by immunohistochemistry
<p>Abstract</p> <p>Background</p> <p>Human breast cancer is classified by gene expression profile into subtypes consisting of two hormone (oestrogen and/or progesterone) receptor-positive types (luminal-like A and luminal-like B) and three hormone receptor-negative types [human epidermal growth factor receptor 2-expressing, basal-like, and unclassified ("normal-like")]. Immunohistochemical surrogate panels are also proposed to potentially identify the molecular-based groups. The present study aimed to apply an immunohistochemical panel (anti-ER, -PR, -ERB-B2, -CK 5/6 and -CK14) in a series of canine malignant mammary tumours to verify the molecular-based classification, its correlation with invasion and grade, and its use as a prognostic aid in veterinary practice.</p> <p>Results</p> <p>Thirty-five tumours with luminal pattern (ER+ and PR+) were subgrouped into 13 A type and 22 B type, if ERB-B2 positive or negative. Most luminal-like A and basal-like tumours were grade 1 carcinomas, while the percentage of luminal B tumours was higher in grades 2 and 3 (Pearson Chi-square P = 0.009). No difference in the percentage of molecular subtypes was found between simple and complex/mixed carcinomas (Pearson Chi-square P = 0.47). No significant results were obtained by survival analysis, even if basal-like tumours had a more favourable prognosis than luminal-like lesions.</p> <p>Conclusion</p> <p>The panel of antibodies identified only three tumour groups (luminal-like A and B, and basal-like) in the dog. Even though canine mammary tumours may be a model of human breast cancer, the existence of the same carcinoma molecular subtypes in women awaits confirmation. Canine mammary carcinomas show high molecular heterogeneity, which would benefit from a classification based on molecular differences. Stage and grade showed independent associations with survival in the multivariate regression, while molecular subtype grouping and histological type did not show associations. This suggests that caution should be used when applying this classification to the dog, in which invasion and grade supply the most important prognostic information.</p
Heat shock protein expression in canine malignant mammary tumours.
Background
Abnormal levels of Heat Shock Proteins (HSPs) have been observed in many human neoplasms including breast cancer and it has been demonstrated that they have both prognostic and therapeutic implications. In this study, we evaluated immunohistochemical expression of HSPs in normal and neoplastic canine mammary glands and confronted these results with overall survival (OS), in order to understand the role of HSPs in carcinogenesis and to establish their potential prognostic and/or therapeutic value.
Methods
Immunohistochemical expression of Hsp27, Hsp72, Hsp73 and Hsp90 was evaluated in 3 normal canine mammary glands and 30 malignant mammary tumours (10 in situ carcinomas, 10 invasive carcinomas limited to local structures without identifiable invasion of blood or lymphatic vessels, 10 carcinomas with invasion of blood or lymphatic vessels and/or metastases to regional lymph nodes). A semi-quantitative method was used for the analysis of the results.
Results
Widespread constitutive expression of Hsp73 and Hsp90 was detected in normal tissue, Hsp72 appeared to be focally distributed and Hsp27 showed a negative to rare weak immunostaining. In mammary tumours, a significant increase in Hsp27 (P < 0.01), Hsp72 (P < 0.05) and Hsp90 (P < 0.01) expression was observed as well as a significant reduction in Hsp73 (P < 0.01) immunoreactivity compared to normal mammary gland tissue. Hsp27 demonstrated a strong positivity in infiltrating tumour cells and metaplastic squamous elements of invasive groups. High Hsp27 expression also appeared to be significantly correlated to a shorter OS (P = 0.00087). Intense immunolabelling of Hsp72 and Hsp73 was frequently detected in infiltrative or inflammatory tumour areas. Hsp90 expression was high in all tumours and, like Hsp73, it also showed an intense positivity in lymphatic emboli.
Conclusion
These results suggest that Hsp27, Hsp72 and Hsp90 are involved in canine mammary gland carcinogenesis. In addition, Hsp27 appears to be implicated in tumour invasiveness and its high immunodetection in invasive tumours is indicative of a poorer clinical outcome
Comparison of Trehalose/Hyaluronic Acid (HA) vs. 0.001% Hydrocortisone/HA Eyedrops on Signs and Inflammatory Markers in a Desiccating Model of Dry Eye Disease (DED)
Background: Dry eye disease (DED) is a multifactorial disease where ocular surface inflammation and damage play key etiological roles. Purpose: To compare a combination of 3% trehalose (T) and 0.15% hyaluronic acid (HA) (Thealoz duo(R), T/HA) with a tear substitute containing 0.001% hydrocortisone (I) and 0.2% HA (Idroflog(R), I/HA), with respect to changes on signs and inflammatory markers in a mouse DED model. Methods: Thirty 12-week-old C57BL/6 mice were exposed in a controlled-environment chamber as a desiccating stress model of DED for 35 days. At day 14 (T1), administration of 5 mu L T or I in the right eye (RE) or NaCl 0.9% in the left eye (LE) started, twice a day. Animals were sacrificed after 7 (T2), 14 (T3), 21 (T4, endpoint) days from the beginning of treatment. Corneal fluorescein staining ratio (Image J), histological and histochemical assessment of ocular surface tissues (goblet cell GC density and characterization -PAS, Alcian blue pH 2.5, pH 1.0, and MUC4 expression-in the superior and inferior conjunctiva), and levels of inflammatory markers HLA-DR, IL-1 beta and TNF-alpha in cornea and conjunctiva were measured. Results: No animal fully recovered from DED signs at the endpoint. Difference between arms was observed at T3 and T4, with T treated eyes showing a higher corneal damage reduction, PAS-positive GC recovery, lower inflammatory marker expression as compared to the I treated ones. Conclusions: Data suggest that 21 days of treatment with T/HA improved signs, GC recovery and inflammatory markers in a DED mouse model, to a greater extent as compared to I/HA. Data suggest that 21 days of treatment with T/HA improved signs, GC recovery and inflammatory markers in a DED mouse model, to a greater extent as compared to I/HA
Diffuse Large B-Cell Lymphoma (DLBCL) in an Aged Raccoon (Procyon lotor)
A 15-years-old, captive, female raccoon (Procyon lotor) was necropsied after a one-week history of apathy and self-isolation. Gross changes consisted of the severe enlargement of the mesenteric lymph node; hepatosplenomegaly with multifocal to coalescing, white tan nodules in the spleen and liver,; and pale kidneys. Histologically, neoplastic CD79Îą-positive lymphocytes effaced the mesenteric lymph node and multifocally infiltrated the spleen, liver, and kidneys, and focally infiltrated the heart. Based on pathological and immunohistochemical findings, as well as the canine-adapted World Health Organization (WHO) diagnostic criteria, a diagnosis of diffuse large B-cell lymphoma (DLBCL) was made
Enteropatia proliferativa da Lawsonia intracellularis nel suino
L\u2019enteropatia proliferativa del suino (proliferative enteropathy - PE), denominata anche ileite, \ue8 causata da Lawsonia intracellularis,
un batterio intracellulare obbligato. La PE \ue8 una patologia a trasmissione oro-fecale che si manifesta soprattutto nella fase
di magronaggio ed \ue8 responsabile di consistenti perdite economiche negli allevamenti intensivi. I danni economici sono causati
dalla riduzione dell\u2019incremento ponderale e dell\u2019indice di conversione dell\u2019alimento e dall\u2019aumento della mortalit\ue0 e dei soggetti
di scarto. La PE \ue8 endemica in numerosi Paesi con prevalenze di aziende ed animali infetti che, in Europa, superano il 90%
e il 40%, rispettivamente. Nel suino, la patologia \ue8 caratterizzata da un ispessimento della mucosa intestinale dovuto alla proliferazione
incontrollata delle cellule delle cripte intestinali accompagnata dall\u2019inibizione, ad opera di L. intracellularis, della maturazione
e della differenziazione delle cellule caliciformi secretorie e delle cellule assorbenti. La conseguenza inevitabile \ue8 la riduzione
dell\u2019assorbimento dei nutrienti e la perdita di aminoacidi e proteine nel lume intestinale, con conseguente diarrea. La
PE si manifesta con due forme cliniche principali: 1) la forma acuta (enteropatia proliferativa emorragica - PHE) che si osserva
principalmente in animali dai 4 ai 12 mesi d\u2019et\ue0, caratterizzata da una diarrea sanguinolenta e da elevata mortalit\ue0 (fino al
50%) e, 2) la forma cronica (adenomatosi intestinale - PIA) che colpisce suini dalle 6 alle 20 settimane d\u2019et\ue0, e il cui sintomo principale
\ue8 rappresentato da una diarrea con feci pastose. Sono state descritte anche altre due forme cliniche a bassa incidenza: a)
l\u2019enterite necrotica (NE), espressione di una forma di enteropatia proliferativa cronica complicata da infezioni secondarie che
esita in un\u2019estesa necrosi coagulativa dell\u2019epitelio intestinale e, b) l\u2019ileite regionale (RI), risultante dalla guarigione delle lesioni
dovute a NE e caratterizzata da deposizione di tessuto di granulazione e ispessimento della tonaca muscolare. La diagnosi indiretta,
che viene generalmente eseguita utilizzando un test ELISA, consente di valutare l\u2019eventuale esposizione dei suini a L. intracellularis,
mentre la diagnosi diretta (realizzata impiegando test biomolecolari qualitativi: PCR o quantitativi: qPCR, immunoistochimica
- IHC) permette di valutare se l\u2019infezione \ue8 in atto. Analogamente a quanto accade per altre forme patologiche
del suino, la semplice messa in evidenza di L. intracellularis nelle feci non rappresenta un criterio diagnostico valido nei confronti
di PE. La corretta procedura diagnostica prevede la quantificazione del numero di microrganismi/grammo di feci e la messa in
evidenza di L. intracellularis all\u2019interno delle lesioni intestinali. La profilassi e il controllo si basano sull\u2019applicazione di rigide misure
di lavaggio e disinfezione che consentono di ridurre la contaminazione ambientale tra un ciclo produttivo e l\u2019altro e l\u2019applicazione
di misure di biosicurezza interna. Particolare attenzione va riservata all\u2019alimentazione, che dovrebbe garantire l\u2019equilibrio
della microflora intestinale tramite un corretto rapporto tra proteina altamente digeribile e frazione di fibra, con il supporto di
integratori probiotici e prebiotici. In una logica di uso consapevole del farmaco, la somministrazione di massa di antibiotici durante
la fase critica del magronaggio dovrebbe essere limitata ai soli gruppi con sintomatologia clinica, implementando invece
la profilassi vaccinale.Porcine Proliferative Enteropathy (PE or ileitis) is an infectious
enteric disease caused by the intracellular pathogen Lawsonia
intracellularis (LI). PE is endemic in many countries and causes
severe economic losses in swine production system worldwide
due to reduction of daily weight gain, reduction of feed
conversion ratio and increase of mortality and swine waste. In
Europe, the prevalence of infected farms and infected animals
is more than 90% and 40%, respectively. In PE, intestinal mucosa
is thickened by uncontrolled proliferation of intestinal crypt
cells while secretory cells and absorbent cells are decreased in
number because LI prevents their maturation. Diarrhea is the
consequence, due to reduced absorption and loss of amino acid
and protein in intestinal lumen. Clinical forms are divided into
acute (proliferative hemorrhagic enteropathy - PHE) and chronic
form (porcine intestinal adenomatosis - PIA). Acute form
affects animals from 4 to 12 weeks of age and is characterized
by high mortality (>50%) and hemorrhagic diarrhea. Chronic
form affects swine of 6-20 weeks of age and is characterized
by pasty diarrhea. Based on morphological findings, two
other forms are reported: necrotic enteritis (NE) and regional
ileitis (RI). The first is a chronic form complicated by secondary
infection that result in coagulative necrosis of intestinal
epithelium. Healing of necrotic enteritis lesions results in both
thickening of muscular layer of intestinal wall and granulation
tissue deposition, both of which are typical findings of RI. Indirect diagnosis (e.g. ELISA) assess the exposure to L. intracellularis
while direct diagnosis (PCR, qPCR, Immunohistochemistry
-IHC) assess the current infection. Effective diagnosis
is obtained comparing quantitation of microorganism/gram
of feces with the detection of L. intracellularis within intestinal
lesion. Prophylaxis and control of proliferative enteropathy are
based on biosecurity measures combined with strict washing
and disinfection measures to reduce environmental contamination.
Proper nutrition management helps to ensure the balance
of intestinal microflora by the use of highly digestible protein,
by correct intake of fiber fraction and with probiotic and
prebiotic supplements. To limit subclinical forms of disease,
vaccination should replace antibiotic treatments which instead
should be reserved only for symptomatic groups of pig
Porcine circovirus type 2 and porcine reproductive and respiratory syndrome virus alone or associated are frequent intralesional detected viruses in porcine respiratory disease complex cases in Northern Italy
Methods: This study aimed to examine the pathological impact of Porcine Circovirus type 2 (PCV2) and Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) through histological and immunohistochemical analysis of 79 cases of Porcine Respiratory Disease Complex (PRDC) collected from 22 farms in Northern Italy. Lung tissue and several lymphoid organ samples were deployed to associate PCV2-positive stain with Circovirus-associated Diseases (PCVD).
Results: The most common lung lesion observed was interstitial pneumonia, alone or combined with bronchopneumonia. By immunohistochemistry, 44 lungs (55.7%) tested positive for PCV2, 34 (43.0%) for PRRSV, 16 (20.3%) for both viruses and in 17 cases (21.5%) neither virus was detected. Twenty-eight out of 44 (63.6%) PCV2-positive cases had lymphoid depletion or granulomatous inflammation in at least one of the lymphoid tissues examined; thus, they were classified as PCV2 Systemic Diseases (PCV2-SD). In the remaining 16 out of 44 cases (36.4%), PCV2-positive lung lesions were associated with hyperplastic or normal lymphoid tissues, which showed PCV2-positive centrofollicular dendritic cells in at least one of the lymphoid tissues examined. Therefore, these cases were classified as PRDC/PCV2-positive. In the PCV2-positive animals, 42.9% of the PCV2-SD cases (12/28) showed immunohistochemistry (IHC) positivity for PRRSV in the lung tissue, while 25.0% of PRDC/PCV2-positive cases (4/16) showed double positivity for PCV2 and PRRSV.
Discussion: In light of the caseload presented in this study, characterized by the high proportion of PCV2-SD cases alongside the overall respiratory symptomatology, it is imperative to emphasize the crucial role of a comprehensive sampling protocol. This is critical to avoid underestimating the harm caused by PCV2 in farms, particularly with respect to the systemic form of the disease. PCV2 and PRRSV remain the primary infections associated with PRDC in Italy that can significantly impact farm health and co-infections in the field can worsen the pathology, thus the selection of appropriate preventive measures is critical
Histochemical Analysis of Herniated Disc Tissue Surgically Removed from 27 Dogs
Hernias of intervertebral discs are a common canine disease that is usually treated surgically. Recently, a histological scoring system for surgically removed canine intervertebral herniated discs has been developed by scoring the lesions of both the anulus fibrosus (AF) and the nucleus pulposus (NP). Since the proportion of AF and NP in the surgical samples may vary, depending on the surgical approach, the aim of this study is to grade separately the lesions in AF and NP and to modify the previously described scoring system adding three parameters: inflammation, mineralization and neovascularization. Possible association of the modified grading system with clinical parameters were statistically assessed. Herniated disc material was collected from 27 dogs. AF was present in 10/27 cases and was classified as grade 2 in 4 cases, grade 3 in 5 cases and grade 4 in 1 case. The NP was present in all 27 cases and was classified as grade 2 in 1 case, grade 3 in 5 cases, grade 4 in 9 cases and grade 5 in 12 cases. A statistically significant association was evidenced between short pre-operative period and higher grade of both the NP and of the AF (P<0.01). Separating the grades of the AF and NP can be useful for a fair assessment of degeneration, and circumventing the limitation of the qualitative and quantitative variability of samples
Bionic for Training: Smart Framework Design for Multisensor Mechatronic Platform Validation
: Home monitoring supports the continuous improvement of the therapy by sharing data with healthcare professionals. It is required when life-threatening events can still occur after hospital discharge such as neonatal apnea. However, multiple sources of external noise could affect data quality and/or increase the misdetection rate. In this study, we developed a mechatronic platform for sensor characterizations and a framework to manage data in the context of neonatal apnea. The platform can simulate the movement of the abdomen in different plausible newborn positions by merging data acquired simultaneously from three-axis accelerometers and infrared sensors. We simulated nine apnea conditions combining three different linear displacements and body postures in the presence of self-generated external noise, showing how it is possible to reduce errors near to zero in phenomena detection. Finally, the development of a smart 8Ws-based software and a customizable mobile application were proposed to facilitate data management and interpretation, classifying the alerts to guarantee the correct information sharing without specialized skills
PCV2 infection in vaccinated conventional gilts inseminated with PCV2b-spiked semen
The present trial investigated the effect of PCV2 vaccination on viremia, virus shedding and viral load in maternal tissues and foetuses of conventional gilts inseminated with PCV2b-spiked semen. Twelve gilts were randomly divided into two groups of six animals each (vaccinated infected, VI; non-vaccinated infected, NVI). Estrus synchronization was followed by artificial insemination (AI) with a single PCV2 negative semen dose supplemented with 0.2 mL of a PCV2b suspension containing 104.4 TCID50/50 \u3bcL (total viral dose: 105 TCID50). Vaginal, nasal and faecal swabs, and blood samples were collected weekly from two days before artificial insemination till the end of the experimental period (55 days post AI; DPAI) and tested by real-time PCR (qPCR) for PCV2; sera were tested for anti- PCV2 antibodies. During necropsy foetal and maternal tissues were collected for qPCR and histopathology. In each of the VI and NVI groups three out of the six gilts were pregnant at 29 DPAI. The VI group showed a significantly lower proportion of PCR-positive swabs: 24.6% VI vs 71.3% NVI. PCV2 clearance was demonstrated by qPCR in lymphoid tissue during the trial in the VI group. Only one foetus was PCV2-positive (in the NVI group) and three amniotic fluids of the NVI group. PCV2 was found in a significantly lower proportion of the placenta of foetuses in the VI group (39%) than the NVI group (77%). The PCV2 vaccine seems to play an active role in reducing virus shedding, tissue viral load and foetoplacental infection
Pathogenetic investigations on the enteric nervous system plexuses of sarda breed sheep with different PrP genotypes following oral experimental scrapie infection
We investigated the ileal myenteric (MPs) and submucosal plexuses (SMPs) of 32 Sarda breed sheep carrying
different PrP genotypes (ARQ/ARQ, ARQ/AHQ, ARQ/ARR, ARR/ARR), which had been orally dosed with
scrapie at 8 months of age and euthanized at definite time intervals post-infection (p.i.)
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