Pathology in Practice.

In collaboration with the American College of Veterinary Pathologists.

Pathology in Practice.

In collaboration with the American College of Veterinary Pathologists.

Pathology in Practice.

In collaboration with the American College of Veterinary Pathologists.

Atypical free-floating iridociliary pigmented epithelial cysts and secondary glaucoma in a caiman (Caiman latirostris).

We describe a case of chronic ocular trauma that resulted in fixed and free-floating, pigmented epithelial iridociliary cysts, inflammation, and secondary glaucoma in a caiman (Caiman latirostris). A 20- to 25-year-old male caiman was presented with phthisis bulbi in the right eye, and congested episcleral vessels, corneal leukoma, disorganized anterior chamber, multifocal anterior synechia, and elevated intraocular pressure in the left eye. Ocular ultrasound of the left eye revealed round structures dispersed in the anterior and posterior chambers and vitreous cavity. Bilateral enucleation was performed, and gross pathology of the left eye revealed multiple pigmented cysts attached to the iris and posterior corneal surface causing marked distortion of the anterior uvea, and free-floating in the vitreous cavity. Histopathology demonstrated heavily pigmented cystic structures of iridociliary epithelium origin carpeting the anterior segment surfaces and causing obstruction of the iridocorneal angles, leading to secondary glaucoma. To the authors' knowledge, this is the first report of iridociliary cysts in wildlife species.

Optimization of the photocatalytic degradation process of aromatic organic compounds applied to mangrove sediment.

Polycyclic aromatic hydrocarbons (PAHs) are part of a class of organic compounds resistant to natural degradation. In this way, heterogeneous photocatalysis becomes useful to degrade persistent organic pollutants, however it can be influenced by environmental variables (i.e.: organic matter) and experimental factors such as: mass of the photocatalyst and irradiation time. The objective of this research was to use a factorial design 2k as a function of the multiple response (MR) to evaluate simultaneously experimental conditions for the photodegradation of polycyclic aromatic hydrocarbons in contaminated mangrove sediment and its application in oil from Potiguar Basin in Brazil. The sediment samples collected in Belmonte city (Southern Bahia state) were contaminated with 0.25 mg kg-1 of Acenaphthene, Anthracene, Benzo[a]Anthracene, Indene[1,2,3cd]pyrene, Dibenzo[ah]anthracene, Benzo[ghi]pyrene. Factors such as mass of the photocatalyst and irradiation time were evaluated in factorial design 22, with triplicate from the central point, to 1g of the PAH contaminated sediment. After performing the experiments, it was found that the best experimental condition for the degradation of all PAHs indicated by MR was the central point (0.5 g of photocatalyst and 12h of irradiation). For such conditions, the half-life of PAHs varied from 3.51 to 9.37 h and the degradation speed constant between 0.0740 to 0.1973 h-1. The comparison of the optimized methodology between photolysis tests and heterogeneous photocatalysis was performed using the Kruskal-Wallis test, which indicated a difference for the reference solution, where heterogeneous photocatalysis was more efficient in the degradation of PAHs. The optimized methodology was apply in samples contaminated with crude oil from Potiguar Basin, no significant difference was observed in the aromatic fraction, using for the Kruskal-Wallis test. Heterogeneous photocatalysis has shown to be a promising remediation technique to remedy aromatic organic compounds in mangrove sediments.

Structural and functional evaluation of the palindromic alanine-rich antimicrobial peptide Pa-MAP2.

Recently, several peptides have been studied regarding the defence process against pathogenic microorganisms, which are able to act against different targets, with the purpose of developing novel bioactive compounds. The present work focuses on the structural and functional evaluation of the palindromic antimicrobial peptide Pa-MAP2, designed based on the peptide Pa-MAP from Pleuronectes americanus. For a better structural understanding, molecular modelling analyses were carried out, together with molecular dynamics and circular dichroism, in different media. Antibacterial activity against Gram-negative and positive bacteria was evaluated, as well as cytotoxicity against human erythrocytes, RAW 264.7, Vero and L6 cells. In silico docking experiments, lipid vesicle studies, and atomic force microscopy (AFM) imaging were carried out to explore the activity of the peptide. In vivo studies on infected mice were also done. The palindromic primary sequence favoured an α-helix structure that was pH dependent, only present on alkaline environment, with dynamic N- and C-terminals that are stabilized in anionic media. Pa-MAP2 only showed activity against Gram-negative bacteria, with a MIC of 3.2 µM, and without any cytotoxic effect. In silico, lipid vesicles and AFM studies confirm the preference for anionic lipids (POPG, POPS, DPPE, DPPG and LPS), with the positively charged lysine residues being essential for the initial electrostatic interaction. In vivo studies showed that Pa-MAP2 increases to 100% the survival rate of mice infected with Escherichia coli. Data here reported indicated that palindromic Pa-MAP2 could be an alternative candidate for use in therapeutics against Gram-negative bacterial infections.

PRELIMINARY REPORT ON THE PUTATIVE ASSOCIATION OF IL10 -3575 T/A GENETIC POLYMORPHISM WITH MALARIA SYMPTOMS.

Only a small percentage of individuals living in endemic areas develop severe malaria suggesting that host genetic factors may play a key role. This study has determined the frequency of single nucleotide polymorphisms (SNPs) in some pro and anti-inflammatory cytokine gene sequences: IL6 (-174; rs1800795), IL12p40 (+1188; rs3212227), IL4 (+33; rs2070874), IL10 (-3575; rs1800890) and TGFb1 (+869; rs1800470), by means of PCR-RFLP. Blood samples were collected from 104 symptomatic and 37 asymptomatic subjects. Laboratory diagnosis was assessed by the thick blood smear test and nested-PCR. No association was found between IL6 (-174), IL12p40 (+1188), IL4 (+33), IL10 (- 3575), TGFb1 (+869) SNPs and malaria symptoms. However, regarding the IL10 -3575 T/A SNP, there were significantly more AA and AT subjects, carrying the polymorphic allele A, in the symptomatic group (c2 = 4.54, p = 0.01, OR = 0.40 [95% CI - 0.17- 0.94]). When the analysis was performed by allele, the frequency of the polymorphic allele A was also significantly higher in the symptomatic group (c2 = 4.50, p = 0.01, OR = 0.45 [95% CI - 0.21-0.95]). In conclusion, this study has suggested the possibility that the IL10 - 3575 T/A SNP might be associated with the presence and maintenance of malaria symptoms in individuals living in endemic areas. Taking into account that this polymorphism is related to decreased IL10 production, a possible role of this SNP in the pathophysiology of malaria is also suggested, but replication studies with a higher number of patients and evaluation of IL10 levels are needed for confirmation.

Effect of yogurt containing polydextrose, Lactobacillus acidophilus NCFM and Bifidobacterium lactis HN019: a randomized, double-blind, controlled study in chronic constipation.

BACKGROUND: Constipation is a frequent complaint and the combination of a prebiotic and probiotics could have a potentially synergic effect on the intestinal transit. The present study therefore aims to investigate the combination of polydextrose (Litesse), L. acidophilus NCFM® and B. lactis HN019 in a yogurt on intestinal transit in subjects who suffer from constipation. METHODS: Patients with constipation were randomly divided into two groups, Control Group (CG) and Treatment Group (TG), and had to eat 180 ml of unflavored yogurt every morning for 14 days. Those in the CG received only yogurt, while the TG received yogurt containing polydextrose, L. acidophilus NCFM (ATCC 700396) and B. lactis HN019 (AGAL NM97/09513). RESULTS: Favourable clinical response was assessed since Agachan score had a significant reduction at the end of the study in both groups and tended to be better in the TG. The subjects in the treatment group also had a shorter transit time at the end of the intervention compared to the control group (p = 0.01). CONCLUSION: The product containing yogurt with polydextrose, B. lactis HN019 and L. acidophilus NCFM® significantly shortened colonic transit time after two weeks in the TG compared to CG and may be an option for treatment of constipation.

The performance of four molecular methods for the laboratory diagnosis of congenital toxoplasmosis in amniotic fluid samples

Introduction Toxoplasmosis may be life-threatening in fetuses and in immune-deficient patients. Conventional laboratory diagnosis of toxoplasmosis is based on the presence of IgM and IgG anti-Toxoplasma gondii antibodies; however, molecular techniques have emerged as alternative tools due to their increased sensitivity. The aim of this study was to compare the performance of 4 PCR-based methods for the laboratory diagnosis of toxoplasmosis. One hundred pregnant women who seroconverted during pregnancy were included in the study. The definition of cases was based on a 12-month follow-up of the infants. Methods Amniotic fluid samples were submitted to DNA extraction and amplification by the following 4 Toxoplasma techniques performed with parasite B1 gene primers: conventional PCR, nested-PCR, multiplex-nested-PCR, and real-time PCR. Seven parameters were analyzed, sensitivity (Se), specificity (Sp), positive predictive value (PPV), negative predictive value (NPV), positive likelihood ratio (PLR), negative likelihood ratio (NLR) and efficiency (Ef). Results Fifty-nine of the 100 infants had toxoplasmosis; 42 (71.2%) had IgM antibodies at birth but were asymptomatic, and the remaining 17 cases had non-detectable IgM antibodies but high IgG antibody titers that were associated with retinochoroiditis in 8 (13.5%) cases, abnormal cranial ultrasound in 5 (8.5%) cases, and signs/symptoms suggestive of infection in 4 (6.8%) cases. The conventional PCR assay detected 50 cases (9 false-negatives), nested-PCR detected 58 cases (1 false-negative and 4 false-positives), multiplex-nested-PCR detected 57 cases (2 false-negatives), and real-time-PCR detected 58 cases (1 false-negative). Conclusions The real-time PCR assay was the best-performing technique based on the parameters of Se (98.3%), Sp (100%), PPV (100%), NPV (97.6%), PLR (∞), NLR (0.017), and Ef (99%). .

In vivo antimicrobial evaluation of an alanine-rich peptide derived from Pleuronectes americanus.

In several organisms, the first barrier against microbial infections consists of antimicrobial peptides (AMPs) which are molecules that act as components of the innate immune system. Recent studies have demonstrated that AMPs can perform various functions in different tissues or physiological conditions. In this view, this study was carried out in order to evaluate the multifunctional activity in vivo of an alanine-rich peptide, known as Pa-MAP, derived from the polar fish Pleuronectes americanus. Pa-MAP was evaluated in intraperitoneally infected mice with a sub-lethal concentration of Escherichia coli at standard concentrations of 1 and 5 mg kg(-1). At both concentrations, Pa-MAPs exhibited an ability to prevent E. coli infection and increase mice survival, similar to the result observed in mice treated with ampicillin at 2 mg kg(-1). In addition, mice were monitored for weight loss. The results showed that mice treated with Pa-MAPs at 1 mg kg(-1) gained 0.8% of body weight during the 72 h of experiment. The same was observed with Pa-MAP at 5 mg kg(-1), which had a gain of 0.5% in body weight during the treatment. Mice treated with ampicillin at 2 mg kg(-1) show a significant weight loss of 5.6% of body weight. The untreated group exhibited a 5.5% loss of body weight. The immunomodulatory effects were also evaluated by the quantification of IL-10, IL-12, TNF-α, IFN-γ and nitric oxide cytokines in serum, but no immunomodulatory activity was observed. Data presented here suggest that Pa-MAP should be used as a novel antibiotic against infection control.

The performance of four molecular methods for the laboratory diagnosis of congenital toxoplasmosis in amniotic fluid samples.

INTRODUCTION: Toxoplasmosis may be life-threatening in fetuses and in immune-deficient patients. Conventional laboratory diagnosis of toxoplasmosis is based on the presence of IgM and IgG anti-Toxoplasma gondii antibodies; however, molecular techniques have emerged as alternative tools due to their increased sensitivity. The aim of this study was to compare the performance of 4 PCR-based methods for the laboratory diagnosis of toxoplasmosis. One hundred pregnant women who seroconverted during pregnancy were included in the study. The definition of cases was based on a 12-month follow-up of the infants. METHODS: Amniotic fluid samples were submitted to DNA extraction and amplification by the following 4 Toxoplasma techniques performed with parasite B1 gene primers: conventional PCR, nested-PCR, multiplex-nested-PCR, and real-time PCR. Seven parameters were analyzed, sensitivity (Se), specificity (Sp), positive predictive value (PPV), negative predictive value (NPV), positive likelihood ratio (PLR), negative likelihood ratio (NLR) and efficiency (Ef). RESULTS: Fifty-nine of the 100 infants had toxoplasmosis; 42 (71.2%) had IgM antibodies at birth but were asymptomatic, and the remaining 17 cases had non-detectable IgM antibodies but high IgG antibody titers that were associated with retinochoroiditis in 8 (13.5%) cases, abnormal cranial ultrasound in 5 (8.5%) cases, and signs/symptoms suggestive of infection in 4 (6.8%) cases. The conventional PCR assay detected 50 cases (9 false-negatives), nested-PCR detected 58 cases (1 false-negative and 4 false-positives), multiplex-nested-PCR detected 57 cases (2 false-negatives), and real-time-PCR detected 58 cases (1 false-negative). CONCLUSIONS: The real-time PCR assay was the best-performing technique based on the parameters of Se (98.3%), Sp (100%), PPV (100%), NPV (97.6%), PLR (âˆz), NLR (0.017), and Ef (99%).

Nylon bands as bone cerclage - experimental study in rabbits and rats / Cinta de náilon como cerclagem óssea - estudo experimental em coelhos e ratos

The aim of this study was to evaluate the biocampatibility of the commercial nylon band used as bone cerclage, comparing with traditional metal cerclage wires. Twenty one rabbits and fifteen rats, healthy and without orthopedics problems were used. A commercial nylon band and a surgical stainless steel wire 0 were used in right (SN) and left (SA) rabbitss hindlimb, respectively. Otherwise, on rats SN was on the left hindlimb, and SA on the right hindlimb. Both experimental species were divided into three groups according to the end of the evaluation period. Clinical, radiographic, macroscopic and histologically evaluation were performed at 30, 60 and 150 days postoperatively. All the animals showed normal limb function differences between groups were not observed at clinical and macroscopic evaluation. On both species, radiographic changes were slightly higher on SN. Histological differences were not observed in both species. Nylon bands do not cause foreign body reaction and other histological changes in rabbits and rats.

O presente trabalho teve como objetivo avaliar a biocompatibilidade da aplicação da cinta de náilon comercial como cerclagem óssea e compará-la com a cerclagem tradicional de fio aço. Foram utilizados 21 coelhos e 15 ratos, sadios e sem alterações no aparelho locomotor. Nos coelhos, uma cinta de náilon comercial foi aplicada no membro pélvico direito (SN) e um fio de aço cirúrgico no 0 no esquerdo (SA). Nos ratos, o SN foi o membro esquerdo e o SA o direito. Ambas as espécies foram separadas em três grupos de acordo com o período final de avaliação. Os animais foram avaliados por exames clínico, radiográfico, macroscópico, e histológico aos 30, 60 e 150 dias pós-implantação. Todos os animais apresentaram função locomotora normal; na avaliação clínica e macroscópica não foram observadas diferenças entre os grupos. Em ambas as espécies, as alterações radiográficas peri-implante foram levemente superiores no implante náilon em relação ao fio de aço. Não foram observadas diferenças na análise microscópica entre os subgrupos náilon e aço, em ambas as espécies. Pode-se concluir que a cinta de náilon não causa reações de corpo estranho nem alterações histológicas no osso normal em ratos e coelhos.

Nylon bands as bone cerclage - experimental study in rabbits and rats / Cinta de náilon como cerclagem óssea - estudo experimental em coelhos e ratos

The aim of this study was to evaluate the biocampatibility of the commercial nylon band used as bone cerclage, comparing with traditional metal cerclage wires. Twenty one rabbits and fifteen rats, healthy and without orthopedics problems were used. A commercial nylon band and a surgical stainless steel wire 0 were used in right (SN) and left (SA) rabbitss hindlimb, respectively. Otherwise, on rats SN was on the left hindlimb, and SA on the right hindlimb. Both experimental species were divided into three groups according to the end of the evaluation period. Clinical, radiographic, macroscopic and histologically evaluation were performed at 30, 60 and 150 days postoperatively. All the animals showed normal limb function differences between groups were not observed at clinical and macroscopic evaluation. On both species, radiographic changes were slightly higher on SN. Histological differences were not observed in both species. Nylon bands do not cause foreign body reaction and other histological changes in rabbits and rats.

O presente trabalho teve como objetivo avaliar a biocompatibilidade da aplicação da cinta de náilon comercial como cerclagem óssea e compará-la com a cerclagem tradicional de fio aço. Foram utilizados 21 coelhos e 15 ratos, sadios e sem alterações no aparelho locomotor. Nos coelhos, uma cinta de náilon comercial foi aplicada no membro pélvico direito (SN) e um fio de aço cirúrgico no 0 no esquerdo (SA). Nos ratos, o SN foi o membro esquerdo e o SA o direito. Ambas as espécies foram separadas em três grupos de acordo com o período final de avaliação. Os animais foram avaliados por exames clínico, radiográfico, macroscópico, e histológico aos 30, 60 e 150 dias pós-implantação. Todos os animais apresentaram função locomotora normal; na avaliação clínica e macroscópica não foram observadas diferenças entre os grupos. Em ambas as espécies, as alterações radiográficas peri-implante foram levemente superiores no implante náilon em relação ao fio de aço. Não foram observadas diferenças na análise microscópica entre os subgrupos náilon e aço, em ambas as espécies. Pode-se concluir que a cinta de náilon não causa reações de corpo estranho nem alterações histológicas no osso normal em ratos e coelhos.

Intoxicação experimental por Kalanchoe Blossfeldiana (crassulaceae) em cães / Experimental intoxication by Kalanchoe Blossfeldiana (crassulaceae) in dogs

Este estudo visou à avaliação clínica e laboratorial da administração oral de Kalanchoe blossfeldiana a cães. Para isso foram utilizados doze cães, adultos, com idade entre dois a cinco anos, sem predileção de sexo ou raça. Os animais receberam extratos de K. blossfeldiana e foram monitorados clínica elaboratorialmente através de hemograma, dosagem sérica de creatinoquinase (CK), alaninoaminotransferase (ALT), fosfatase alcalina (FA) e eletrocardiograma acerca das alterações provocadas pelos princípios tóxicos da planta. A avaliação dos animais não revelou nenhuma alteração clínica e/ou laboratorial, estando os valores das enzimas CK, ALT, FA e hemograma dentro dos valores normais. No exame eletrocardiográfico todos os animais apresentaram normalidade de ritmo (arritmia sinusal), eixo cardíaco e nas medidas das ondas, não sendo evidenciado nenhum indício de necrose ou isquemia cardíaca. Os resultados mostraram que o extrato de K. bloosfeldiana não produz um quadro de intoxicação em cães nas condições em que foi desenvolvido o experimento.

The purpose of this study was to evaluate the clinical and laboratorial aspects of the oral administration of Kalanchoe blossfeldiana in dogs. Twelve dogs, from two to five years of age, without breed or sex preferences, were used. The animals were inoculated with the K. blossfeldiana extract and monitored by complete blood count (CBC), serum biochemical analysis of creatinokinase (CK), alanine aminotransferase (ALT), alkaline phosfatase (FA) and electrocardiographic exam for any cardio-toxic effect from the extract. No significant changes were identified by clinical examination, CBC, blood chemistry or EKG exams, with all results within standard values. The electrocardiographic exam of all the animals presented normal rhythm (sinus arrhythmia), cardiac axle and wave measures, without necrose or cardiac ischemia indication. These results suggested that K. blossfeldiana extract do not cause acute intoxication in dogs within the scope of this experiment.

Intoxicação experimental por Kalanchoe Blossfeldiana (crassulaceae) em cães / Experimental intoxication by Kalanchoe Blossfeldiana (crassulaceae) in dogs

Este estudo visou à avaliação clínica e laboratorial da administração oral de Kalanchoe blossfeldiana a cães. Para isso foram utilizados doze cães, adultos, com idade entre dois a cinco anos, sem predileção de sexo ou raça. Os animais receberam extratos de K. blossfeldiana e foram monitorados clínica elaboratorialmente através de hemograma, dosagem sérica de creatinoquinase (CK), alaninoaminotransferase (ALT), fosfatase alcalina (FA) e eletrocardiograma acerca das alterações provocadas pelos princípios tóxicos da planta. A avaliação dos animais não revelou nenhuma alteração clínica e/ou laboratorial, estando os valores das enzimas CK, ALT, FA e hemograma dentro dos valores normais. No exame eletrocardiográfico todos os animais apresentaram normalidade de ritmo (arritmia sinusal), eixo cardíaco e nas medidas das ondas, não sendo evidenciado nenhum indício de necrose ou isquemia cardíaca. Os resultados mostraram que o extrato de K. bloosfeldiana não produz um quadro de intoxicação em cães nas condições em que foi desenvolvido o experimento.(AU)

The purpose of this study was to evaluate the clinical and laboratorial aspects of the oral administration of Kalanchoe blossfeldiana in dogs. Twelve dogs, from two to five years of age, without breed or sex preferences, were used. The animals were inoculated with the K. blossfeldiana extract and monitored by complete blood count (CBC), serum biochemical analysis of creatinokinase (CK), alanine aminotransferase (ALT), alkaline phosfatase (FA) and electrocardiographic exam for any cardio-toxic effect from the extract. No significant changes were identified by clinical examination, CBC, blood chemistry or EKG exams, with all results within standard values. The electrocardiographic exam of all the animals presented normal rhythm (sinus arrhythmia), cardiac axle and wave measures, without necrose or cardiac ischemia indication. These results suggested that K. blossfeldiana extract do not cause acute intoxication in dogs within the scope of this experiment.(AU)

The environmental influence of Light Coke, phosphoric acid, and citric acid on elastomeric chains.

AIM: The purpose of this study was to evaluate the in vitro effect of Light Coke, phosphoric acid, and citric acid on the force decline pattern of two types of elastomeric chains. METHODS AND MATERIALS: One hundred sixty gray colored elastomeric chain modules, 80 Chainette (GAC) and 80 Sunburst (GAC) chains, were divided into four groups for immersion into Light Coke, phosphoric acid, citric acid, and artificial saliva. The initial stretched force of the elastomeric chains ranged from 220 gf to 250 gf. During the experiment, all elastomeric chains were kept immersed in artificial saliva at 37 masculineC (pH 6.24) to simulate the oral environment. To simulate daily consumption of Light Coke, the elastomeric chains were immersed in the solutions twice a day for 15 minutes. The control group was kept immersed in artificial saliva continuously with no further treatment. Force (gf) was measured with a Dial-Type dynamometer (Dentaurum), initially (baseline), 24 hours, 7, 14, and 21 days. Data were analyzed using the Tukey test at a 5% level of probability. RESULTS: A statistically significant reduction on the force produced by the elastomeric chains was seen at different time points. The greatest reduction in force occurred in the first 24 hours (p=0.01). CONCLUSION: Force decay of the Chainette elastomeric chains were lower than the Sunburst (p<0.05). The immersion treatments caused no statistically significant difference in force for either chain module (p>0.05). CLINICAL SIGNIFICANCE: Good elastomeric chain properties are necessary for effective tooth movement, and knowledge of force-decay rates is an important factor in achieving the best orthodontic outcome.