Imidazolium salts as alternative compounds to control diseases caused by plant pathogenic bacteria.

AIMS: To evaluate the inhibitory effect of five structurally different imidazolium salts on the in vitro growth of plant pathogenic bacteria that belong to divergent taxonomic genera as well as their ability to reduce the severity of common bacterial blight of common bean caused by Xanthomonas axonopodis pv. phaseoli and bacterial speck of tomato caused by Pseudomonas syringae pv. tomato. METHODS AND RESULTS: Growth inhibition of Xanthomonas, Pseudomonas, Erwinia, Pectobacterium and Dickeya strains by imidazolium salts was assessed in vitro by radial diffusion on agar medium and by ressazurin reduction in liquid medium. The reduction of common bacterial blight and bacterial speck symptoms and the area under de disease progress curves were determined by spraying two selected imidazolium salts on healthy plants 48 h prior to inoculation with virulent strains of the bacterial pathogens. All imidazolium salts inhibited the growth of all plant pathogenic bacteria when tested by radial diffusion on agar medium. The strength of inhibition differed among imidazolium salts when tested on the same bacterial strain and among bacterial strains when tested with the same imidazolium salt. In liquid medium, most imidazolium salts presented the same minimum inhibitory concentration (MIC) and minimum bactericidal concentration values (200 µmol l-1 ), the most notable exception of which was the MIC (at least 1000 µmol l-1 ) for the dicationic MImC10 MImBr2 . The imidazolium salts C16 MImBr and C16 MImCl caused significant reductions in the severity of common bacterial blight symptoms when compared with nontreated plants. CONCLUSION: Imidazolium salts inhibit the in vitro growth of plant pathogenic bacteria and reduce plant disease symptoms to levels comparable to an authorized commercial antibiotic product. SIGNIFICANCE AND IMPACT OF THE STUDY: New compounds exhibiting broad-spectrum antibacterial activity with potential use in agriculture were identified.

Tissue eosinophilia in oral intraepithelial neoplasia as a probable indicator of invasion.

OBJECTIVE: To investigate the presence of eosinophils in oral intraepithelial neoplasia (OIN) and oral squamous cell carcinoma (OSCC) lesions and its relation to invasion. SUBJECTS AND METHODS: Ninety-nine oral biopsies were selected and subdivided into the following: OIN-1(16 cases), OIN-2 (18 cases), OIN-3 (17 cases), microinvasive OSCC (10 cases), non-metastatic OSCC (22 cases) and metastatic OSCC (16 cases). The tissue eosinophilia was evaluated histologically in slices stained with haematoxylin and eosin. RESULTS: Eosinophil distribution was associated with diagnosis severity (p < .01). A significant difference was found between OIN-3 or microinvasive OSCC and non-metastatic or metastatic OSCC. Stromal invasion threshold was 7 eos/10 high power field (hpf) (96.1% specificity and 62.5% sensitivity). Eosinophils were absent in OIN-1; in OIN-2, two cases were positive. In OIN-3, five cases showed tissue eosinophilia, four of which had ≥3 eos/hpf or ≥7 eos/10 hpf. Three cases were suspected of invasion; two had a previous history of OSCC with elevated eosinophil infiltrate. In microinvasive OSCC, the four positive cases presented ≥3 eos/hpf and ≥7 eos/10 hpf. Although not significantly different, non-metastatic invasive OSCC had a higher number of cases (68.2%) with ≥22 eos/10 hpf contrasting with 50% in metastatic OSCC. CONCLUSION: Our results suggest that eosinophils can be considered an indicator of invasion in OIN, helping in cases of difficult diagnosis.

Differential expression of immune-modulatory molecule HLA-E in non-neoplastic and neoplastic lesions of the thyroid.

Human leukocyte antigen (HLA)–E is a non-classical molecule of the histocompatibility complex that functions as one of the main ligands of the Natural Killer (NK) cell inhibitory receptor CD94/NKG2A and inhibits its potent cytotoxic activity. Due to the important role of NK cells in combating neoplasm, we hypothesized that the differential expression of HLA-E could favor the progression of heterogeneous thyroid tumors.Using an immunohistochemistry technique in 143 biopsies of thyroid tumors, including benign and malignant neoplasms and goiters, we evaluated the expression of HLA-E among various tumor types and its association with the clinicopathological factors of diseases. We verified high HLA-E expression in all types of neoplastic tumors, although no significant differences between the groups were found. Low expression was observed in 95 percent of the goiter samples, showing significant differences between neoplastic and non-neoplastic lesions. Furthermore, a significant result was found with regard to the tumor size, with high HLA-E expression being related to smaller tumors. Therefore, our data suggest that an increase in HLA-E may be associated with the establishment of thyroid neoplasms, with either benign or malignant features.

Expression of human leucocyte antigen-G primarily targets affected skin of patients with psoriasis.

BACKGROUND: The nonclassical human leucocyte antigen (HLA)-G molecule has been well recognized as a tolerogenic molecule and few studies have evaluated the role of the molecule in inflammatory cutaneous autoimmune diseases. OBJECTIVES: To evaluate the expression of HLA-G in skin specimens of patients with psoriasis and to analyse its correlation with epidemiological and clinical variables. METHODS: Thirty untreated patients with psoriasis and 32 healthy individuals were enrolled. Immunohistochemistry was applied to identify HLA-G expression in formalin-fixed paraffin-embedded cutaneous skin biopsies. RESULTS: Soluble and membrane-bound HLA-G expression was detected in 30 (90%) of the skin specimens from patients presenting clinical and histopathological features of psoriasis. Although infiltrating lymphomononuclear cells of the dermis exhibited HLA-G expression, the epidermis was primarily targeted. HLA-G expression was also observed in 27% (three of 11) of the specimens that exhibited no clinical and histopathological features of psoriasis (nonaffected areas). In contrast, skin specimens obtained from healthy individuals exhibited no HLA-G expression (P < 0·0001). The intensity of HLA-G expression was not associated with type I/II psoriasis, Psoriasis Area and Severity Index score or clinical forms. CONCLUSIONS: As the HLA-G molecule was consistently expressed in affected and, to a lesser extent, in nonaffected areas of untreated patients with psoriasis, irrespective of the severity of the clinical variants, one may hypothesize that the presence of HLA-G may be responsible, at least in part, for the regulation of autoimmune effector cells.

Effects of ethyl-cyanoacrylate and octyl-cyanoacrylate on experimental corneal lesions in rabbits / Efeitos do etil-cianoacrilato ou do octil-cianoacrilato sobre lesões corneais experimentais em coelhos

The use of ethyl-cyanoacrylate and octyl-cyanoacrylate were clinically and histopathologically compared on the corneas of 36 rabbits after lamellar keratectomy (standardized diameter and depth). The animals were distributed into two groups, one for each type of adhesive. From each group, six subgroups were histopathologically evaluated on the 3rd, 7th, 14th, 21st, 30th, and 60th day post-operative. General (daily) and ophthalmic (days 0, 1, 3, 5, 7, 14, 21, 30, 44, and 60) evaluations clinically indicated that there were significant differences for the variables water intake, attitude, blepharitis, corneal edema, and fluorescein test. The adhesive permanence time for octyl-cyanoacrylate (17.22 days) was greater than that for ethyl-cyanoacrylate (7.66 days). With respect to the histopathological evaluation, corneal epithelization and collagen organization occurred without severe complications. However, treatment with ethyl-cyanoacrylate led to a moderate inflammatory reaction in the initial phases. With octyl-cyanoacrylate, re-epithelization and collagen organization proceeded more slowly with a discrete inflammatory reaction in the initial phases. From clinical and histopathologic points of view, octyl-cyanoacrylate showed advantages over ethyl-cyanoacrylate, whereas wound healing was achieved in both groups without major complications.

Comparou-se o uso do etil-cianoacrilato e do octil-cianoacrilato em córneas de 36 coelhos após ceratectomia lamelar (diâmetro e profundidade padronizados). Os animais foram distribuídos em dois grupos, segundo o tipo de adesivo, e redistribuídos em seis subgrupos com três animais cada, para as avaliações histológicas aos 3, 7, 14, 21, 30 e 60 dias de pós-operatório. As avaliações clínicas gerais (diárias) e as oftálmicas (dias 0, 1, 3, 5, 7, 14, 21, 30, 44 e 60), indicaram diferença entre os dois grupos, quanto ao consumo de água, atitude, blefarite, edema da córnea e teste da fluoresceína. O Tempo de permanência, sobre o leito corneal, do adesivo octil-cianoacrilato (17,22 dias), foi maior que o do etil-cianoacrulato (7,66 dias). A histopatologia, para ambos os grupos, mostrou que a re-epitelização e a organização do colágeno ocorreram sem graves intercorrências. O grupo tratado com o etil-cianoacrilato apresentou, nas fases iniciais, reação inflamatória mais evidente que o tratado com octil-cianoacrilato. Neste, a re-epitelização e a organização do colágeno ocorreram mais lentamente e com reação inflamatória discreta. Sob os pontos de vista clínico e de avaliação histológica simples, os resultados mostraram vantagens do octil-cianoacrilato, entretanto, a cicatrização da córnea ocorreu em ambos os grupos.

Anti-eosinophilic effect of Lafoensia pacari in toxocariasis.

We previously reported the anti-inflammatory activity of Lafoensia pacari extract in Toxocara canis infection, a model of systemic IL-5-dependent eosinophil migration. In the present study, we describe the kinetics of the anti-inflammatory activity of L. pacari extract and compare it with dexamethasone. T. canis-infected mice were submitted to different treatment protocols and the cells present in bronchoalveolar space and peritoneal cavity were collected at the end of each treatment period. The results showed that L. pacari extract effectively inhibited eosinophil migration only when the treatment was initiated before the peak of eosinophil migration (1st to 18th; 12th to 18th and 12th to 24th day post-infection). When eosinophil migration was established, administration of L. pacari extract had no effect on it (treatment 18th to 24th day post-infection). Dexamethasone was effective in inhibiting eosinophil migration in all periods studied. We suggest that L. pacari extract can potentially be a natural alternative treatment of eosinophilic diseases.

Anti-inflammatory activity of quercetin and isoquercitrin in experimental murine allergic asthma.

OBJECTIVE: Eosinophils and cytokines are implicated in the pathogenesis of allergic diseases. In the present study, we investigate the anti-inflammatory effect of quercetin and isoquercitrin in a murine model of asthma. METHODS: BALB/c mice were immunized (ovalbumin/aluminum hydroxide, s. c.), followed by two intranasal ovalbumin challenges. From day 18 to day 22 after the first immunization, the mice received daily gavages of isoquercitrin (15 mg/kg) or quercetin (10 mg/kg). Dexamethasone (1 mg/kg, s. c.) was administered as a positive control. Leucocytes were analyzed in bronchoalveolar lavage fluid (BALF), blood and pulmonary parenchyma at 24 h after the last ovalbumin challenge. Interleukin-5 (IL-5) was analyzed in BALF and lung homogenates. RESULTS: In animals receiving isoquercitrin or quercetin, eosinophil counts were lower in the BALF, blood and lung parenchyma. Neutrophil counts in blood and IL-5 levels in lung homogenate were lower only in isoquercitrin-treated mice. No alterations in mononuclear cell numbers were observed. CONCLUSION: Quercetin and isoquercitrin are effective eosinophilic inflammation suppressors, suggesting a potential for treating allergies.

Impact of MK886 on eosinophil counts and phenotypic features in toxocariasis.

Experimental toxocariasis was used as a model of eosinophil migration. Mice inoculated with 200 Toxocara canis eggs were treated with the leukotriene inhibitor MK886 (1 mg/kg/day). Eosinophils were counted in peripheral blood (PB), peritoneal cavity (PC) and bronchoalveolar lavage fluid (BALF) samples on post-infection days 3, 6, 12, 18, 24 and 36. Eosinophil expression of Mac-1 and VLA-4 was analysed in PB and PC samples. We found that T. canis infection induced systemic eosinophilia from post-infection day 3, peaking on days 6, 12 and 24 in PB, PC and BALF samples respectively. Eosinophilia was more pronounced in PB and PC samples than in BALF samples, and MK886 downregulated eosinophilia to varying degrees in the different sample types. In PB and PC samples, T. canis infection caused early upregulation of Mac-1 with late changes in the VLA-4 profile, whereas MK886 had opposite effects. The distinct time-dependent eosinophilia peaks and differential involvement of leukotrienes in integrin expression demonstrate that, despite the systemic eosinophilia triggered by T. canis infection, inflammatory responses vary by compartment.

Quantitative cell-cycle protein expression in oral cancer assessed by computer-assisted system.

The knowledge of cell-cycle control has shown that the capacity of malignant growth is acquired by the stepwise accumulation of defects in specific genes regulating cell growth. Histologic diagnosis might be improved by a quantitative evaluation of more specific diagnosis biomarkers, which could help to precisely identify pre-malignant and malignant oral lesions. The aim of the present study is to evaluate whether computer-based quantitative assessment of p53, PCNA and Ki-67 immunohistochemical expression, could be used clinically to foresee the risk of oral malignant transformation. This retrospective study was carried out in ninety-five oral biopsies, 27 were classified as fibrous inflammatory hyperplasia, 40 as leukoplakia and 28 as oral squamous cell carcinoma. Sixteen out of the 40 leukoplakia were diagnosed as non-dysplastic leukoplakia, the other 24 being dysplastic leukoplakia, of which 50.0% were classified as moderate to severe dysplasia. Comparison of the four groups of oral tissues showed significant rises in p53 and Ki-67 positivity index, which increased steadily in the order benign, pre-malignant, and malignant. In contrast, it was not possible to relate higher PCNA levels with pre-malignant and malignant oral lesions. We therefore conclude that PCNA immunohistochemistry expression is probably an inappropriate marker to identify oral carcinogenesis, whereas joint quantitative evaluation of p53 and Ki-67, appears to be useful as a tumor marker, providing a pre-diagnostic estimate of the potential for cell-cycle deregulation of the oral proliferate status.

Association of tumor necrosis factor a-2 and a-8 microsatellite alleles with human papillomavirus and squamous intraepithelial lesions among women in Brazil.

Infection with oncogenic human papillomavirus (HPV) is considered to be the major risk factor for cervical cancer. Tumor necrosis factor (TNF) is a pluripotent cytokine that plays an important role in inhibiting the action of microbial agents, and TNF microsatellite polymorphisms have been associated with several diseases, including cancer and viral infections. This study analyzed the associations between TNFa to -e microsatellite polymorphisms and the severity of squamous intraepithelial lesions (SIL), according to the presence of the oncogenic HPV16 and HPV18 types. Samples from 146 HPV-positive women with low-grade SIL (LSIL) and high-grade SIL (HSIL) and samples from 101 healthy women were studied. TNF microsatellite polymorphism typing and HPV detection and typing were performed using PCR-amplified DNA hybridized with sequence-specific primers. Data were analyzed by Fisher's exact test using the GENEPOP software. Significant associations were observed between LSIL and the TNFa-8 allele (4/166; P = 0.04), as well as between TNFa-2 with HPV18 only (16/44; P = 0.002) and TNFa-2 with HPV18 coinfection with HPV16 (16/44; P = 0.001). Patients exhibiting the TNFa-2 allele and harboring HPV18, in the presence or absence of coinfection with HPV16, had an increased risk of HSIL occurrence (13/38; P = 0.04; 5/10; P = 0.04) compared to patients with other HPV types. These results suggest that the TNFa-8 allele is associated with increased susceptibility to the occurrence of LSIL and that despite the presence of a high TNF-alpha production allele, the ability of HPV18 to resist the inhibitory effects of TNF-alpha may contribute to the occurrence of infection and consequently to HSIL in women with cervical HPV18 infection.

Immune regulatory effect of pHSP65 DNA therapy in pulmonary tuberculosis: activation of CD8+ cells, interferon-gamma recovery and reduction of lung injury.

A DNA vaccine based on the heat-shock protein 65 Mycobacterium leprae gene (pHSP65) presented a prophylactic and therapeutic effect in an experimental model of tuberculosis. In this paper, we addressed the question of which protective mechanisms are activated in Mycobacterium tuberculosis-infected mice after immune therapy with pHSP65. We evaluated activation of the cellular immune response in the lungs of infected mice 30 days after infection (initiation of immune therapy) and in those of uninfected mice. After 70 days (end of immune therapy), the immune responses of infected untreated mice, infected pHSP65-treated mice and infected pCDNA3-treated mice were also evaluated. Our results show that the most significant effect of pHSP65 was the stimulation of CD8+ lung cell activation, interferon-gamma recovery and reduction of lung injury. There was also partial restoration of the production of tumour necrosis factor-alpha. Treatment with pcDNA3 vector also induced an immune stimulatory effect. However, only infected pHSP65-treated mice were able to produce significant levels of interferon-gamma and to restrict the growth of bacilli.

Langerhans' cell count and HLA class II profile in cervical intraepithelial neoplasia in the presence or absence of HIV infection.

OBJECTIVES: The progression of immunosuppression in human immunodeficiency virus (HIV)+ women has been correlated with elevated incidence of squamous intraepithelial lesions (SIL), probably indicating the role of local immune milieu. In this study, we analysed S100, and HLA class II molecule expression in cervical biopsies according to HIV status, to the severity of SIL and to human papillomavirus (HPV) type. METHODS: Biopsies from 34 HIV+ and 44 HIV- patients with normal cervix or low- or high-grade SIL were studied. Langerhans' cells (LC) (S100), HLA class II and HLA-DQ molecules were evaluated by immunohistochemistry. HPV detection was performed using polymerase chain reaction (PCR). For statistical analysis Mann-Whitney (P< or =0.05) and Spearman test were used. RESULTS: Epithelial S100 and HLA class II density were significantly increased with the severity of lesion (P=0.032; P=0.005). Epithelial S100+ increased in HPV+ (P=0.038), and HLA class II density decreased in HPV 16+ (P=0.035) or 18+ (P<0.0001) samples. HIV infection was associated with increased stromal S100+ (P=0.0005) and decreased HLA class II densities (P=0.0001). Decreased stromal S100+ was observed in women with CD4<500 cells/microl (P=0.050). Among HIV+ patients with SIL, the lowest S100 and epithelial HLA class II densities were detected in women with CD4<200 cells/microl (P=0.045). CONCLUSIONS: After the establishment of AIDS, increased numbers of immature LCs and a reduction in HLA class II occurred, possibly turning the cervical milieu more favourable to HPV persistence. HPV 16 and 18 infections may interfere with the antigen presenting activity, possibly as an evasion mechanism.

Influencia do laser de baixa potencia nos niveis das proteinas plasmaticas de coelhos / Low power laser influence on the plasmatic proteins levels in rabbits

Os efeitos do laser de baixa potencia parecem nao se limitar ao local da aplicacao. Eles podem, por meio dos mediadores metabolicos, chegar a areas mais distantes do corpo promovendo efeitos sistemicos. O objetivo destre trabalho foi analisar as mudancas dos niveis de proteinas sericas, como uma forma de determinar um dos efeitos sistemicos do laser de baixa potencia. Trinta e seis coelhos machos foram induzidos a um processo inflamatorio articular no joelho da pata direita com terebinthina comum (Tc). Foram divididos aleatoriamente em 3 grupos, cada um com 12 animais, e subdivididos em grupos de 6 animais para receber as densidades energeticas diferenciadas de 3.4 J.cm e 8 J.cm. O grupo exerimental agudo (GEA) iniciou o laser 48 horas apos a injecao da Tc o grupo experimental cronico (GEC) recebeu laser 5 dias depois da inducao da inflamacao e o grupo ontrole, que nao recebeu tratamento e foi subdividido, por sua vez, em 6 animais (GCA) que seguiram o mesmo esquema do GEA e outros 6 (GCC), O esquema do GEC. Tambem foi colhido o sangue de outros 30 animais normais para obter um grupo-padrao. Usou-se um laser com comprimento de onda de 830 nm e potencia de 77 mW, aplicado durante 7 dias, uma sessao a cada 24 horas. O sangue foi processado para eletroforse. O modelo experimental foi adequado, uma vez que os animais dos grupos sob inducao da inflamacao se comportaram diferentemente do grupo-padrao. Nos animais com infamacao induzida, foram observadas as seguintes alteracoes diminuicao acentuada da albumina no GCC, aumento da-globulina no GEA e da-globulina nos GCA e GCC e elevacao da y-globulina no GCC. Poucas diferencas foram encontradas quando comparadas as duas doses de laser rmpregadas (3,4 e 8 J.cm ). Os resultados indicam haver efeito sistematico sobre as proteinas plasmaticas promovido pelo laser de baixa potencia nesse modelo de infamacao articular induzida

Estudo experimental da resposta terapêutica em abcessos sub-cutâneos provocados por injeçäo de terebintina em ratos / Experimental research of the therapeutical response in subcutaneous abscesses in mice caused by ingestion of tupentine

Os autores estudaram experimentalmente a açäo antiinflamatória de Belladonna e Silicea em ratos nos quais se provocou abcesso subcutâneo com injeçäo de terebintina.

Massive intracranial immature teratoma. Report of a case with polyhidramnios and intense pelvic pain.

A case of massive intracranial immature teratoma in a female stillborn is reported. She was the product of the second pregnancy of a 25-year-old healthy woman. The pregnancy was unremarkable until the 25th week of gestation when the mother noticed a rapid enlargement of her abdomen and intense pelvic pain. Because of the pain, a cesarean section was indicated, and a stillborn weighing 2750g with macrocephaly was delivered. The cranial contents weighed 1350g and showed a huge tumoral mass with only a rim of normal brain. A histologic diagnosis of immature teratoma was made. Massive intracranial teratomas are rare tumors and their occurrence in intrauterine life is even rarer. Their histogenesis is unknown, and there is no explanation for their continuous growth during embryogenesis. A prenatal diagnosis of this rare condition can be made by ultrasound, computed tomography, or magnetic resonance imaging.

Influence of misoprostol, a synthetic prostaglandin E1 analog, on the healing of colonic anastomoses in rats.

PURPOSE: This study was designed to examine the influence of misoprostol, a synthetic prostaglandin E1 analog, on the healing of colonic anastomoses in rats, with particular regard to changes in collagen levels at the site of the anastomoses and their histopathologic aspects. METHODS: Sixty rats were submitted to resection and anastomosis of the colon, and divided at random into two groups. The test group received misoprostol intragastrically (200 micrograms/kg body weight), twice daily, from the day of operation until sacrifice. Controls received 0.9 percent NaCl. The animals were sacrificed on the third, seventh, or fourteenth postoperative day, and the results of the histopathologic analyses and hydroxyproline concentrations were compared. RESULTS: Our results show that misoprostol administration increased the hydroxyproline concentration on the fourteenth postoperative day without interfering in the inflammatory response (P < 0.05). CONCLUSION: Misoprostol interferes with the balance between the synthesis and degradation of collagen, resulting in an elevation of collagen levels by the fourteenth postoperative day without influencing the inflammatory response.

Effects of early postnatal malnutrition and chlordiazepoxide on experimental aversive situations.

In order to study the lasting consequences of brain changes caused by early malnutrition, rats were fed a protein-deficient diet from birth until 49 days of age and a balanced diet from day 50 to day 70. At 49 and 70 days of age, independent groups of animals were tested in the locomotor activity, step-down inhibitory avoidance, and flinch-jump nociceptive tests. Also, at 49 days of age, malnourished and control rats were sacrificed in order to evaluate the weight of brain regions. Malnourished rats had lower body and brain weights (telencephalon and brain stem) than control rats. Malnourished rats also showed less locomotor activity at the beginning of the test session, lower flinch and jump thresholds, and longer step-down latencies than control animals. Chlordiazepoxide (5 mg/kg, IP) shortened step-down latency of well-nourished rats, but was ineffective in malnourished rats. These and previously reported results indicate that early protein malnutrition causes long-lasting impairment of neuronal systems underlying emotional behavior.

Poliploidia em Tumor de Wilms / Polyploidia in tumor of Wilms

Os autores analisam citogeneticamente celulas derivadas de um tumor de Wilms objetivando estabelecer correlacao entre poliploidia e histologia desfavoravel. O exame citogenetico do caso foi realizado por metodo diretocom analise de 20 metafases, que revelou poliploidia em 100% das celulas analisadas, com variacao de 60 a 69 cromossomos. O resultado histologico foi do tipo blastemal anaplasico. Os dados sao consistentes com outros da literatura realizados tanto com exame citogenetico como atraves de analise de DNA por citometria de fluxo, que associam poliploidia com histologia desfavoravel e confirmam o exame citogenetico como um elemento importante na avaliacao do prognostico desta neoplasia .