Ag Nanoparticles/α-Ag2WO4 Composite Formed by Electron Beam and Femtosecond Irradiation as Potent Antifungal and Antitumor Agents.

The ability to manipulate the structure and function of promising systems via external stimuli is emerging with the development of reconfigurable and programmable multifunctional materials. Increasing antifungal and antitumor activity requires novel, effective treatments to be diligently sought. In this work, the synthesis, characterization, and in vitro biological screening of pure α-Ag2WO4, irradiated with electrons and with non-focused and focused femtosecond laser beams are reported. We demonstrate, for the first time, that Ag nanoparticles/α-Ag2WO4 composite displays potent antifungal and antitumor activity. This composite had an extreme low inhibition concentration against Candida albicans, cause the modulation of α-Ag2WO4 perform the fungicidal activity more efficient. For tumor activity, it was found that the composite showed a high selectivity against the cancer cells (MB49), thus depleting the populations of cancer cells by necrosis and apoptosis, without the healthy cells (BALB/3T3) being affected.

Lactococcus garvieae outbreaks in Brazilian farms Lactococcosis in Pseudoplatystoma sp. - development of an autogenous vaccine as a control strategy.

This study evaluated the control of streptococcosis outbreaks in Brazil, isolated from diseased sorubim and identified as Lactococcus garvieae by genetic sequencing. This report determined the potential for lactococcosis control in sorubim Pseudoplatystoma sp. with two vaccines: an aqueous-based, whole-cell inactivated vaccine (bacterin) and an oil-adjuvanted bacterin. Their efficacy was evaluated at 30 days post-vaccination (d.p.v.) by challenge with L. garvieae, and the antibody production response at 15, 30 and 60 d.p.v. and the non-specific immune response were compared amongst treatments. High protection levels (P < 0.05) were achieved with the oil-adjuvanted vaccine with a relative percentage survival value of 81.7% at 30 d.p.v. Additionally, the oil-adjuvanted vaccine increased the immunogenicity of the bacterin as indicated by greater agglutination antibody titres from 15 until 60 d.p.v. This is the first report of a positive effect of vaccine administration on the specific immunity of sorubim, and the study showed that a specific antibody plays an important role in sorubim defence against lactococcosis because the innate immune responses were similar in all of the studied animals. These results demonstrated that oil-adjuvanted vaccine can be an effective alternative for the protection of sorubim from L. garvieae disease.

Visual perception of multilocular radiolucent mandibular lesions quantified by morphometric analysis.

OBJECTIVE: The aim of this study was to analyse the visual perceptions of different experts with respect to multilocular radiolucent lesions in circumstances when the diagnosis is either known or unknown. METHODS: 6 radiographs of ameloblastomas (AMELs), keratocystic odontogenic tumours (KOTs), central giant cell lesions (CGCLs) and myxomas (MIXs) were analysed by 16 dental experts [stomatologists/oral surgeons (SS) and dental radiologists (R)]. They delimited the lesions prior to having knowledge of the diagnosis (T1) and after 30 days, when they were aware of the histopathological results (T2). For each image, the following morphometric parameters were calculated: area (A), perimeter (P) and shape factor (SF); after image subtraction procedures (T1 - T2), the exclusive area (EA) of the non-overlapping delimited region was also calculated. RESULTS: For both groups, the T2 area was larger than the T1, although the EA of the SS group was higher than that of the R group independently of the type of lesion. The SF from the SS group was greater than that from the R group, and at T2 the SF values were higher for both groups. AMELs and MIXs showed larger SF and A values; the SS group tended to have the greatest changes in the delimitations of the lesions at T2. CONCLUSIONS: The methodology allowed us to quantify differences in the spatial perceptions of professionals. The knowledge of the diagnosis and the expertise of examiners influenced the examiner's perception of the limits of the lesions independently of the actual biological behaviour of the lesion.

Detection of Paracoccidioides brasiliensis by PCR in biopsies from patients with paracoccidioidomycosis: correlation with the histopathological pattern.

Paracoccidioidomycosis, a systemic mycosis, is rarely diagnosed in its initial phase and can remain latent for up to 40 years. Although PCR is sensitive for the identification of Paracoccidioides brasiliensis (Pb) in different samples, no study using paraffin-embedded human tissue has been published. The size of the amplicon, the fixation method and the time of the storage may affect the reaction. Recently the more sensitive Primer-Extension-Preamplification (PEP)-Nested-PCR has been used for amplification of small samples. Our aims were to detect Pb in paraffin embedded biopsies using (PEP)-Nested-PCR and to correlate the data with histopathological parameters. Analyses were carried out in 107 biopsies from tegument, lymph node, lung and tongue. The fungal DNA was detected in 29.9% of the biopsies by (PEP)-nested-PCR against 5% of Nested-PCR. The positivity correlated with numbers of fungi and fungal viable cells, and there was no correlation with the granuloma pattern.

Development of an open case-based decision-support system for diagnosis in oral pathology.

BACKGROUND: Making diagnoses in oral pathology are often difficult and confusing in dental practice, especially for the less-experienced dental student. One of the most promising areas in bioinformatics is computer-aided diagnosis, where a computer system is capable of imitating human reasoning ability and provides diagnoses with an accuracy approaching that of expert professionals. This type of system could be an alternative tool for assisting dental students to overcome the difficulties of the oral pathology learning process. This could allow students to define variables and information, important to improving the decision-making performance. However, no current open data management system has been integrated with an artificial intelligence system in a user-friendly environment. Such a system could also be used as an education tool to help students perform diagnoses. The aim of the present study was to develop and test an open case-based decision-support system. METHODS: An open decision-support system based on Bayes' theorem connected to a relational database was developed using the C++ programming language. The software was tested in the computerisation of a surgical pathology service and in simulating the diagnosis of 43 known cases of oral bone disease. The simulation was performed after the system was initially filled with data from 401 cases of oral bone disease. RESULTS: The system allowed the authors to construct and to manage a pathology database, and to simulate diagnoses using the variables from the database. CONCLUSION: Combining a relational database and an open decision-support system in the same user-friendly environment proved effective in simulating diagnoses based on information from an updated database.

Differential gene expression assessed by cDNA microarray analysis in breast cancer tissue under tamoxifen treatment.

Our purpose was to identify tamoxifen (TAM) responsive genes after 30 days of TAM treatment in tumor tissues obtained from women with breast cancer using microarray expression analysis. In our study, we identified 12 candidates to be considered as tamoxifen-modulated genes. Among them, we selected two candidates the TEGT BI-1 (testis enhanced gene transcript Bax Inhibitor-1) and the CD63 gene in order to further confirm their differential expression under tamoxifen effects. We observed that both were down-regulated in tumor tissues of patients during TAM treatment. TEGT is able to inhibit the expression of Bax, which is known to promote apoptosis. On the other hand, CD63 encodes a cell membrane protein and it seems to be involved in mechanisms of platelet activation, cell adhesion and cell motility. We therefore hypothesize that TAM would be able to modulate tumor growth by down-regulating genes involved in mechanisms such as cell cycle control, tumor invasion and metastasis.

Gene expression profiles reveal that DCN, DIO1, and DIO2 are underexpressed in benign and malignant thyroid tumors.

To investigate the molecular events involved in the pathogenesis and/or progression of thyroid tumors, we compared the gene expression profiles of three thyroid carcinoma cell lines, which represent major tumor subtypes of thyroid cancer and normal thyroid tissue. Using cDNA array methodology, we investigated the expression of 1807 open reading frame expressed sequence tags (ORESTES), selected from head and neck tumor libraries generated through the Brazilian Human Cancer Project-LICR/FAPESP. We found that 505 transcripts were differentially expressed in the thyroid carcinoma cell lines. Using a more stringent criterion, transcripts underexpressed or overexpressed more than fivefold in 1 of 3 or 3 of 3 carcinoma cell lines, a list of 55 ESTs were detected. Five candidate genes were further validated by quantitative polymerase chain reaction (qPCR) in an independent set of 52 thyroid tumors and 22 matched normal thyroid tissues. DCN was found underexpressed in a high percentage of the follicular thyroid adenomas, follicular thyroid carcinomas, and follicular variant of papillary thyroid carcinomas. DIO1 and DIO2 were underexpressed in nearly all papillary thyroid carcinomas. These genes not only could help to better define a tumor signature for thyroid tumors, but may, in part, also become useful as potential targets for thyroid tumor treatment.

Canine paracoccidioidomycosis.

Paracoccidioidomycosis (PCM) is a severe disease caused by the dimorphic fungus Paracoccidioides brasiliensis, which is characterized by granulomatous pulmonary and systemic lesions, affecting mainly men between 20 and 60 years of age. Reports of PCM disease in animals are rare, but the disease has been described in armadillos. On the other hand, PCM infection of domestic and wild animals detected by serological or cutaneous tests in the absence of apparent disease has been frequently reported. We present here the case of a female adult Doberman that developed cervical lymphadenomegaly. Histopathological examination of a cervical biopsy specimen revealed active PCM, with an epithelioid, granulomatous inflammation containing numerous yeast-like, multiple budding fungal forms. The diagnosis of PCM was confirmed by immunohistochemistry using a specific antibody anti-gp43 and by nested PCR using primers for the amplification of the gp43 gene region. This is the first report of PCM disease occurring in a dog, an animal that has been shown to play an important role in the natural history of North American blastomycosis.

FTY720 impairs necrosis development after ischemia-reperfusion injury.

Ischemia-reperfusion (IR) injury is a common early feature that contributes to graft damage by impairing resident cell function. Our previous results showed that IR injury impaired renal function, by causing extensive tubular necrosis and increasing MHC class II and ICAM-1 molecule expression by mesangial cells (MC). MCs are likely candidates to come into close contact with immune cells such as monocytes or lymphocytes. It has been suggested that under inflammatory circumstances, there is increased MC expression of MHC class II, of adhesion molecules (such as ICAM-1), of cytokines receptors, and of molecules associated with cellular death (apoptosis). The immunosuppressive properties of FTY720 have been shown in clinical and experimental situations. It has also been shown to be protective against IR injury in rats. We sought to evaluate the role of FTY720 in a murine IR model by measuring renal function, tubular necrosis, and surface molecule expression by cultured mesangial cells. Intravenous administration of FTY720 (1 mg/kg) immediately before IR induction did not improve the short-term (24 hours) outcome of renal function or reduced MHC class II and ICAM-1 surface molecule expression. However, there was a decreased percentage of tubular necrosis in mice treated with FTY720 (51.3% +/- 1.6%) compared with vehicle-treated mice (66% +/- 5.5%). These results suggest a protective role of FTY720 in an IR injury model. More studies are required to identify the mechanisms involved in the protective activity of FTY720 in the IR injury model.

The Th1 /Th2 immune-type response of the recurrent aphthous ulceration analyzed by cDNA microarray.

BACKGROUND: The reduced ability to activate oral tolerance plays a role in the pathogenesis of some gastrointestinal inflammatory diseases. This activation may reflect a preferential reduction of a T-helper (Th)2- or Th3-type response. In recurrent aphthous ulceration (RAU), genetic and environmental factors may contribute to low tolerance, permitting a cytotoxic reaction against the oral epithelium. The cytokine profile has not permitted the definition of RAU as resulting from enhanced Th1 or Th2 responses. A cDNA microarray study would allow the identification of differentially expressed genes and provide a basis for classification of the immune response. METHODS: The cDNA from 29 samples of aphthae and from 11 samples of normal mucosa from aphthae-free volunteers were hybridized on microarray membranes with 1176 genes. RESULTS: Forty-one differentially expressed genes were identified, and a higher expression level of the Th1 gene cluster in RAU was found. CONCLUSIONS: Microarrays permitted us definition of the gene expression profile of the lesion and identify an increased Th1 activity in RAU lesions.

Estrogen and selective estrogen receptor modulator regulation of insulin-like growth factor binding protein 5 in the rat uterus.

Insulin-like growth factor (IGF) binding protein 5 (IGFPB-5) is abundant in the uterus and is implicated in the sex steroid-induced growth of this tissue. The purpose of this study was to investigate the potential for modulation of the action of IGFPB-5 at the uterus level in the rat by estrogen and selective estrogen receptor modulators (SERMs). One hundred and twenty adult rats, 2-3 months of age, were included. Among them 100 animals were ovariectomized 4 days prior to the use of drugs for 48 days. Rats were divided into six groups: non-ovariectomized (group 1); ovariectomized (group 2); and those receiving conjugated equine estrogens, 50 microg/day (group 3), tamoxifen 250 microg/day (group 4), raloxifene 3 mg/kg (group 5) and toremifene 2.5 mg/kg (group 6). Total RNA was isolated from the uterus and IGFBP-5 mRNA levels were assessed by the semiquantitative reverse transcription polymerase chain reaction (RT-PCR). Our results demonstrate that conjugated equine estrogens were able to up-regulate mRNA levels of the IGFBP-5 gene, while oophorectomy alone as well as associated with hormone therapy such as tamoxifen, raloxifene and toremifene resulted in down-regulation of uterine IGFBP-5 gene expression. The up-regulation of IGFBP-5 expression induced by estrogens suggests that, in vivo, the uterotrophic effects of estrogen replacement therapy are mediated, at least inpart, by the IGF pathways. On the other hand, the use of SERMs (tamoxifen, raloxifene and toremifene) was associated with severe down-regulation of this gene at the transcription level.

Estrogen regulation of uterine genes in vivo detected by complementary DNA array.

INTRODUCTION: In the present study, our aim was to identify differentially expressed genes involved in estrogen actions at the endometrium level in rats. METHODS: Thirty adult rats were ovariectomized four days prior to drug administration for 48 days. Rats were divided in 2 groups: I, control and II, conjugated equine estrogens (CCE). Total RNA was isolated from uterus, and differential expression was analyzed by array technology and RT-PCR. RESULTS: A total of 32 candidate genes were shown to be upregulated or downregulated in groups I or II. Among them, differential expression was already confirmed by RT-PCR for IGFBP5, S12, c-kit, and VEGF, genes whose expression was up regulated during CCE therapy, and casein kinase II and serine kinase expression was the same level in both groups. CONCLUSION: We have demonstrated that cDNA array represents a powerful approach to identify key molecules in the estrogens therapy. A number of the candidates reported here should provide new markers that may contribute to the detection of target estrogen receptor. This information may also aid the development of new approaches to therapeutic intervention.

p53 In biopsies of oral squamous cell carcinoma. A comparative study with a malignancy grading system.

The relationship between the histological grading of malignancy and p53 protein expression was studied in 40 biopsies of oral squamous cell carcinomas. An immunohistochemical analysis was carried out using the streptavidin method preceded by a treatment with citric acid in a microwave oven. All cases were classified according to the histological malignancy grading system proposed by Anneroth et al. (Scandinavian Journal of Dental Research 1987, 95, 229-249). The expression of p53 was found in 62.5% of the carcinomas studied. Positivity of p53 staining showed a correlation with the histological grade of malignancy and with the degree of keratinisation, nuclear polymorphism and number of mitoses.

[Computer-assisted image processing for quantifying histopathologic variables in the healing of colonic anastomosis in dogs]. / Aplicação do processamento de imagens por computador na quantificação des variáveis histopathológicas de reparação tecidual de anastomoses colocólicas em cães.

BACKGROUND: The authors present the experimental results of the computerized quantifying of tissular structures involved in the reparative process of colonic anastomosis performed by manual suture and biofragmentable ring. The quantified variables in this study were: oedema fluid, myofiber tissue, blood vessel and cellular nuclei. MATERIAL AND METHODS: An image processing software developed at Laboratório de Informática Dedicado à Odontologia (LIDO) was utilized to quantifying the pathognomonic alterations in the inflammatory process in colonic anastomosis performed in 14 dogs. The results were compared to those obtained through traditional way diagnosis by two pathologists in view of counterproof measures. The criteria for these diagnoses were defined in levels represented by absent, light, moderate and intensive which were compared to analysis performed by the computer. RESULTS: There was significant statistical difference between two techniques: the biofragmentable ring technique exhibited low oedema fluid, organized myofiber tissue and higher number of alongated cellular nuclei in relation to manual suture technique. CONCLUSION: The analysis of histometric variables through computational image processing was considered efficient and powerful to quantify the main tissular inflammatory and reparative changing.