Metallothionein immunoexpression in non-syndromic and syndromic keratocystic odontogenic tumour

Background: To commpare the metallothionein (MT) immunoexpression in non-syndromic and syndromic keratocystic odontogenic tumour (KOT), to correlate MT with cellular proliferation, and to evaluate the influence of inflammation in MT. Material and Methods: Fourteen cases of KOT were submitted to imm unohistochemistry for MT and Ki-67 analysis. The lesions were grouped according to their grade of inflammation, and statistical analysis was performed. Results: MT was higher in non-syndromic KOT than in syndromic KOT (p<0.05). No statistical difference in Ki- 67 could be identified; however, an inverse correlation was observed between MT and Ki-67 in both lesions. When analysing inflammation, non-syndromic KOT showed no differences in either MT or Ki-67. Conclusions: The MT imm unophenotype of syndromic KOT was different from non-syndromic KOT. MT might not be involved in the proliferation control of both KOT. MT and Ki-67 imm unoexp ressions proved to be unaffected by inflammation in non-syndromic KOT

Inhibitory Effect of Cold Atmospheric Plasma on Chronic Wound-Related Multispecies Biofilms

The presence of microbial biofilms in the wounds affects negatively the healing process and can contribute to therapeutic failures. This study aimed to establish the effective parameters of cold atmospheric plasma (CAP) against wound-related multispecies and monospecies biofilms, and to evaluate the cytotoxicity and genotoxicity of the protocol. Monospecies and multispecies biofilms were formed by methicillin-resistant Staphylococcus aureus (MRSA), Pseudomonas aeruginosa and Enterococcus faecalis. The monospecies biofilms were grown in 96 wells plates and multispecies biofilm were formed on collagen membranes. The biofilms were exposed to helium CAP for 1, 3, 5 and 7 min. In monospecies biofilms, the inhibitory effect was detected after 1 min of exposure for E. faecalis and after 3 min for MRSA. A reduction in P. aeruginosa biofilm’s viability was detected after 7 min of exposure. For the multispecies biofilms, the reduction in the overall viability was detected after 5 min of exposure to CAP. Additionally, cytotoxicity and genotoxicity were evaluated by MTT assay and static cytometry, respectively. CAP showed low cytotoxicity and no genotoxicity to mouse fibroblastic cell line (3T3). It could be concluded that He-CAP showed inhibitory effect on wound-related multispecies biofilms, with low cytotoxicity and genotoxicity to mammalian cells. These findings point out the potential application of CAP in wound care

Cold Atmospheric Pressure Plasma Is Effective against P. gingivalis (HW24D-1) Mature Biofilms and Non-Genotoxic to Oral Cells

The effects of helium cold atmospheric pressure plasma (He-CAPP) jet on Porphyromonas gingivalis (HW24D-1) biofilm, on human gingival fibroblasts (HGF) and human gingival keratinocytes (OBA-9) were assessed. Standardized suspension of P. gingivalis was obtained, and biofilms were grown anaerobically for 48 h. After exposition to He-CAPP, the biofilm viability was evaluated by XTT assay. HGF were grown at 37 °C, in an CO2 chamber in DMEM, while OBA-9 cells were cultured in keratinocyte serum-free medium. After 24 h, plates were exposed to He-CAPP for 1 to 7 min. Plasma was generated using a commercial AC power supply with amplitude modulated signal (voltage amplitude of 20 kVp-p, frequency of 31.0 kHz and duty cycle of 22%). The corresponding discharge power was 0.6W at He flow rate of 1 L/min. DNA damage was accessed by static cytometry. Data were analyzed by GraphPad Prism (p < 0.05). Significant reductions in P. gingivalis viability in relation to non-treated groups were detected (p < 0.0001), directly proportional to exposure time. Treated groups were slightly aneuploid after 5- and 7-min treatment in HGF, and for 3 min in OBA-9 cells, with 1.2 DNA index mean. Helium cold atmospheric pressure plasma jet showed inhibitory effect on P. gingivalis mature biofilm and was not genotoxic for epithelial gingival cells and human oral fibroblasts

Análise quantitativa das lesões cardíacas na cardiomiopatia chagásica crônica canina

Lesions observed in chronic chagasic cardiopathy frequently produce electrocardiographic alterations and affect cardiac function. Through a computerized morphometrical analysis we quantified the areas occupied by cardiac muscle, connective and adipose tissues in the right atrium of dogs experimentally infected with Trypanosoma cruzi. All of the infected dogs showed chronic myocarditis with variable reduction levels of cardiac muscle, fibrosis and adipose tissue replacement. In the atrial myocardium of dogs infected with Be78 and Be62 cardiac muscle represented 34 and 50%, fibrosis 28 and 32% and adipose tissue 38 and 18%, respectively. The fibrosis observed was both diffuse and focal and mostly intrafascicular, either partially or completely interrupting the path of muscle bundles. Such histological alterations probably contributed to the appearance of electrocardiographic disturbances verified in 10 out 11 dogs which are also common in human chronic chagasic cardiopathy. Fibrosis was the most important microscopic occurrence found since it produces rearrangements of collagen fibers in relation to myocardiocytes which causes changes in anatomical physiognomy and mechanical behavior of the myocardium. These abnormalities can contribute to the appearance of cardiac malfunction, arrythmias and congestive cardiac insufficiency as observed in two of the analyzed dogs. Strain Be78 caused destruction of atrial cardiac muscle higher than that induced by strain Be62.As lesões observadas na cardiopatia chagásica crônica frequentemente produzem alterações eletrocardiográficas e afetam a função cardíaca. Através de uma análise morfométrica computadorizada nós quantificamos as áreas ocupadas por músculo cardíaco, tecido conjuntivo fibroso e tecido adiposo no átrio direito de cães experimentalmente infectados pelo Trypanosoma cruzi. Todos os cães infectados apresentaram miocardite crônica fibrosante com graus variáveis de redução de músculo cardíaco, fibrose e substituição por tecido adiposo. No miocárdio atrial dos cães infectados pelas cepas Be78 e Be62 foram observados 34 e 50% de músculo cardíaco, 28 e 32% de fibrose e, 38 e 18% de tecido adiposo, respectivamente. A fibrose observada era tanto difusa quanto focal e, principalmente intrafascicular interrompendo total ou parcialmente o percurso dos feixes musculares. Tais alterações histológicas provavelmente contribuiram para o surgimento dos distúrbios eletrocardiográficos verificados em 10 dos 11 cães estudados e que são comuns na cardiopatia chagásica crônica humana. De todos os achados microscópicos encontrados, a fibrose foi a mais importante por produzir rearranjos na fibras colágenas em relação aos miocardiócitos, modificando a fisionomia anatômica e o comportamento mecânico do miocárdio. Tais anormalidades estruturais podem contribuir para o surgimento à disfunção cardíaca, arritmias e à insuficiência cardíaca congestiva como verificado em dois cães analisados. A cepa Be78 produziu uma destruição de músculo cardíaco atrial estatisticamente superior à induzida pela cepa Be62

Colite necrohemorrágica causada por Entamoeba histolytica em um cão

Background: Intestinal amebiasis with morphological lesions and clinical manifestations is uncommon in dogs. The disease is caused by the protozoan Entamoeba histolytica, which is commonly observed in its natural hosts, humans and some non-human primates. It is occasionally found in the company of animals, usually associated with contact with infected humans. Thus, the objective here is to describe a case of necro-hemorrhagic colitis caused by E. histolytica in a dog infected with the canine distemper virus, in order to characterize the epidemiological and clinicopathological aspects of the disease.Case: An adult, mixed-breed bitch displaying anorexia and ataxia was referred to the veterinary hospital for treatment. Clinical evaluation showed a cachectic animal with 12% dehydration, ocular discharge, and bilateral purulent nasal dis-charge. A clinical diagnosis of distemper was made, and treatment was instituted. The dog’s signs progressed to walking in circles, aimlessly, with lethargy and blindness. After three days of the onset of neurological signs, the dog developed diarrhea with hematochezia. With no improvement noted, we elected to euthanize the dog. At necropsy, edema was present in subcutaneous tissues, and the lungs had yellow areas in the cranio-ventral portions, which the court was flowing purulent discharge. In the large intestine, segmental distention of the distal portion of the descending colon was observed. The segment was approximately 15 cm in length and consisted of irregular reddish areas. There was also slight thickening of the wall with edematous mucosa containing blood clots, fibrin, and multiple areas of ulceration. Microscopically showed necro-hemorrhagic colitis associated with rounded structures, approximately 15 μm in size, containing abundant eosinophilic cytoplasm that was slightly granular or vacuolated. They also contained nuclei and nucleoli that were central or slightly eccentric. These organisms were consistent with amoeba trophozoites. There was also demyelinating encephalitis associated with malacia, corpuscular intranuclear eosinophilic inclusions and / or intracytoplasmic inclusions in ependymal cells, astrocytes, and gemistocytes, characteristic of infection with canine distemper virus. Using immunohistochemistry with polyclonal anti-E. histolytica antibodies in the dilution of 1:1000, trophozoites were immunomarked, confirming the suspected amebiasis.Discussion: The diagnosis of intestinal amebiasis was based on clinical signs and by morphological characteristics on gross and microscopic examination, and was confirmed as E. histolytica by immunohistochemistry. Limited information on theepidemiology and pathological findings of infection with Entamoeba sp. has been reported in the literature, as it is relatively uncommon in pets. Affected animals are usually asymptomatic, but immunosuppression caused by canine distemper virus may have triggered the clinical manifestations of the disease in this dog. Enteritis due Entamoeba sp. should be considered in dogs with chronic weight loss and bloody diarrhea. It should also be included in the differential diagnoses for weight loss and hemorrhagic gastroenteritis, such as canine parvovirus, canine adenovirus 1, Pythium insidiosum, and Giardia sp

Quantitative analysis of cardiac lesions in chronic canine chagasic cardiomyopathy.

Lesions observed in chronic chagasic cardiopathy frequently produce electrocardiographic alterations and affect cardiac function. Through a computerized morphometrical analysis we quantified the areas occupied by cardiac muscle, connective and adipose tissues in the right atrium of dogs experimentally infected with Trypanosoma cruzi. All of the infected dogs showed chronic myocarditis with variable reduction levels of cardiac muscle, fibrosis and adipose tissue replacement. In the atrial myocardium of dogs infected with Be78 and Be62 cardiac muscle represented 34 and 50%, fibrosis 28 and 32% and adipose tissue 38 and 18%, respectively. The fibrosis observed was both diffuse and focal and mostly intrafascicular, either partially or completely interrupting the path of muscle bundles. Such histological alterations probably contributed to the appearance of electrocardiographic disturbances verified in 10 out 11 dogs which are also common in human chronic chagasic cardiopathy. Fibrosis was the most important microscopic occurrence found since it produces rearrangements of collagen fibers in relation to myocardiocytes which causes changes in anatomical physiognomy and mechanical behavior of the myocardium. These abnormalities can contribute to the appearance of cardiac malfunction, arrhythmias and congestive cardiac insufficiency as observed in two of the analyzed dogs. Strain Be78 caused destruction of atrial cardiac muscle higher than that induced by strain Be62

Influence of inflammation on parasitism and area of experimental amoebic liver abscess: an immunohistochemical and morphometric study

The influence of inflammation on the number of trophozoites and on the murine amoebic liver abscess area following infection with Entamoeba histolytica and E. dispar was evaluated. Immunohistochemistry and digital morphometry were used to identify and quantify the trophozoites, neutrophils, macrophages, and lesions. Positive correlation was observed between the number of trophozoites and inflammatory cells. A significant decrease in parasitism and inflammation in groups treated with dexamethasone was observed. The scarceness or absence of trophozoites in the treated groups suggest the importance of the inflammatory response in the production of amoebic hepatic abscesses in spite of the inherent virulence of the parasite being decisive in the establishment of the lesion

IgG Induced by Vaccination With Ascaris suum Extracts Is Protective Against Infection

Human ascariasis has a global and cosmopolitan distribution, and has been characterized as the most prevalent neglected tropical disease worldwide. The development of a preventive vaccine is highly desirable to complement current measures required for this parasitic infection control and to reduce chronic childhood morbidities. In the present study, we describe the mechanism of protection elicited by a preventive vaccine against ascariasis. Vaccine efficacy was evaluated after immunization with three different Ascaris suum antigen extracts formulated with monophosphoryl lipid A (MPLA) as an adjuvant: crude extract of adult worm (ExAD); crude extract of adult worm cuticle (CUT); and crude extract of infective larvae (L3) (ExL3). Immunogenicity elicited by immunization was assessed by measuring antibody responses, cytokine production, and influx of tissue inflammatory cells. Vaccine efficacy was evaluated by measuring the reductions in the numbers of larvae in the lungs of immunized BALB/c mice that were challenged with A. suum eggs. Moreover, lung physiology and functionality were tested by spirometry to determine clinical efficacy. Finally, the role of host antibody mediated protection was determined by passive transfer of serum from immunized mice. Significant reductions in the total number of migrating larvae were observed in mice immunized with ExL3 61% (p &lt; 0.001), CUT 59% (p &lt; 0.001), and ExAD 51% (p &lt; 0.01) antigens in comparison with non-immunized mice. For the Ascaris antigen-specific IgG antibody levels, a significant and progressive increase was observed with each round of immunization, in association with a marked increase of IgG1 and IgG3 subclasses. Moreover, a significant increase in concentration of IL-5 and IL-10 (pre-challenge) in the blood and IL-10 in the lung tissue (post-challenge) was induced by CUT immunization. Finally, ExL3 and CUT-immunized mice showed a marked improvement in lung pathology and tissue fibrosis as well as reduced pulmonary dysfunction induced by Ascaris challenge, when compared to non-immunized mice. Moreover, the passive transfer of specific IgG antibodies from ExL3, CUT, and ExAD elicited a protective response in naïve mice, with significant reductions in parasite burdens in lungs of 65, 64, and 64%, respectively. Taken together, these studies indicated that IgG antibodies contribute to protective immunity