Hyaluronidases and hyaluronan synthases expression is inversely correlated with malignancy in lung/bronchial pre-neoplastic and neoplastic lesions, affecting prognosis

We collected a series of 136 lung/bronchial and 56 matched lung parenchyma tissue samples from patients who underwent lung/bronchial biopsies and presented invasive carcinoma after lung surgery. The lung/bronchial samples included basal cell hyperplasia, squamous metaplasia, moderate dysplasia, adenomatous hyperplasia, severe dysplasia, squamous cell carcinoma and adenocarcinoma. Matched lung parenchyma tissue samples included 25 squamous cell carcinomas and 31 adenocarcinomas. Immunohistochemistry was performed to analyze for the distribution of hyaluronidase (Hyal)-1 and −3, and hyaluronan synthases (HAS)-1, −2, and −3. Hyal-1 showed significantly higher expression in basal cell hyperplasia than in moderate dysplasia (P=0.01), atypical adenomatous hyperplasia (P=0.0001), or severe dysplasia (P=0.03). Lower expression of Hyal-3 was found in atypical adenomatous hyperplasia than in basal cell hyperplasia (P=0.01) or moderate dysplasia (P=0.02). HAS-2 was significantly higher in severe dysplasia (P=0.002) and in squamous metaplasia (P=0.04) compared with basal cell hyperplasia. HAS-3 was significantly expressed in basal cell hyperplasia compared with atypical adenomatous hyperplasia (P=0.05) and severe dysplasia (P=0.02). Lower expression of HAS-3 was found in severe dysplasia compared with squamous metaplasia (P=0.01) and moderate dysplasia (P=0.01). Epithelial Hyal-1 and −3 and HAS-1, −2, and −3 expressions were significantly higher in pre-neoplastic lesions than in neoplastic lesions. Comparative Cox multivariate analysis controlled by N stage and histologic tumor type showed that patients with high HAS-3 expression in pre-neoplastic cells obtained by lung/bronchial biopsy presented a significantly higher risk of death (HR=1.19; P=0.04). We concluded that localization of Hyal and HAS in lung/bronchial pre-neoplastic and neoplastic lesions was inversely related to malignancy, which implied that visualizing these factors could be a useful diagnostic procedure for suspected lung cancer. Finalizing this conclusion will require a wider study in a randomized and prospective trial.

Role of the extracellular matrix in variations of invasive pathways in lung cancers

Among the most common features of highly invasive tumors, such as lung adenocarcinomas (AD) and squamous cell carcinomas (SqCC), is the massive degradation of the extracellular matrix. The remarkable qualitative and quantitative modifications of hyaluronidases (HAases), hyaluronan synthases (HAS), E-cadherin adhesion molecules, and the transforming growth factor β (TGF-β) may favor invasion, cellular motility, and proliferation. We examined HAase proteins (Hyal), HAS, E-cadherin, and TGF-β profiles in lung AD subtypes and SqCC obtained from smokers and non-smokers. Fifty-six patients, median age 64 years, who underwent lobectomy for AD (N = 31) and SqCC (N = 25) were included in the study. HAS-1, -2 and -3, and Hyal-1 and -3 were significantly more expressed by tumor cells than normal and stroma cells (P < 0.01). When stratified according to histologic types, HAS-3 and Hyal-1 immunoreactivity was significantly increased in tumor cells of AD (P = 0.01) and stroma of SqCC (P = 0.002), respectively. Tobacco history in patients with AD was significantly associated with increased HAS-3 immunoreactivity in tumor cells (P < 0.01). Stroma cells of SqCC from non-smokers presented a significant association with HAS-3 (P < 0.01). Hyal, HAS, E-cadherin, and TGF-β modulate a different tumor-induced invasive pathway in lung AD subgroups and SqCC. HAases in resected AD and SqCC were strongly related to the prognosis. Therefore, our findings suggest that strategies aimed at preventing high HAS-3 and Hyal-1 synthesis, or local responses to low TGF-β and E-cadherin, may have a greater impact in lung cancer prognosis.

Effect of ischemia preconditioning on renal ischemia/reperfusion injury in rats

To study the effect of ischemia preconditioning (IP) on renal ischemia/reperfusion (I/R)-associated functional injury and expression of renal adhesion molecules in rats.

The ischemia preconditioning plan adopted in this experiment involved renal warm ischemia for 6 min. and blood flow for 4 min., repeated four times. The Wistar rat kidneys used for warm ischemia preconditioning were subjected to 60 min of renal warm ischemia followed by reperfusion. The rat kidneys with ischemia/reperfusion were compared with the ischemia preconditioning group to observe rat renal function and changes in the expression of renal adhesion molecules ICAM-1, P--Selectin, and E-Selectin.

The expression of rat renal adhesion molecules (ICAM-1, P-Selectin, and E-Selectin) with ischemia preconditioning was significantly lower than that of the ischemia/reperfusion group. Serum creatinine was significantly lower than that in the ischemia/reperfusion group after 48 hours.

Ischemia preconditioning has a protective effect on renal function. Reduced expression of renal adhesion molecules is likely a mechanism involved in the observed protection. .

Molecular & phenotypic characterization of Staphylococcus epidermidis in implant related infections.

Background & objectives: The discrimination between the Staphylococcus epidermidis colonizing the deep seated indwelling devices and those which are mere commensals has always been a challenge for the clinical microbiologist. This study was aimed to characterize the S. epidermidis isolates obtained from device related infection for their phenotypic and molecular markers of virulence and to see whether these markers can be used to differentiate the pathogenic S. epidermidis from the commensals. Methods: Fifty five S. epidermidis isolates from various device related infections such as endophthalmitis following intra-ocular lens (IOL) implantation, intravascular (IV) catheter related sepsis and orthopaedic implant infections, were studied for slime production, biotyping, antibiotic sensitivity; and mec A and ica positivity by the recommended procedures. Results: Twenty three (41.8%) isolates were multi-drug resistant, 26 (65.2%) were slime producers, 30 (54.5%) were adherent, 23 (41.8%) possessed the intercellular adhesin (ica) gene, and 28 (50.9%) harboured the mec A gene. Biotypes I and III were the commonest, most members of which were multi- drug resistant. Twenty two (73.3%) of the 30 adherent bacteria were slime producers as opposed to only 4 (16%) of the 25 non-adherent bacteria (P<0.001). A vast majority i.e. 21 (91.3%) of the 23 ica positive organisms were adherent to artificial surfaces in contrast to only 9 (28.1%) of the 32 non-ica positive organisms (P<0.001). Twenty (86.9%) of the 23 ica positive bacteria were slime producers, as opposed to only 6 (18.7%) of the 32 ica negative bacteria (P<0.001). Of the 23 multi-drug resistant isolates, 19 (82.6%) carried the mec A gene. Interpretation & conclusions: The present findings showed that ica AB and mec A were the two important virulence markers of S. epidermidis in implant infections and slime was responsible for the sessile mode of attachment on the devices.

Immune expression of E-cadherin and α, β and γ-Catenin adhesion molecules and prognosis for upper urinary tract urothelial carcinomas

INTRODUCTION: Cell adhesion molecules (CAM) are required for maintaining a normal epithelial phenotype, and abnormalities in CAM expression have been related to cancer progression, including bladder urothelial carcinomas. There is only one study that correlates E-cadherin and α-, β- and γ-catenin expression with prognosis of upper tract urothelial carcinomas. Our aim is to study the pattern of immune expression of these CAMs in urothelial carcinomas from the renal pelvis and ureter in patients who have been treated surgically. Our goal is to correlate these expression levels and characteristics with well-known prognostic parameters for disease-free survival. MATERIALS AND METHODS: We evaluated specimens from 20 patients with urothelial carcinomas of the renal pelvis and ureter who were treated with nephroureterectomy or ureterectomy between June 1997 and January 2007. CAM expression was evaluated by immunohistochemistry in a tissue microarray and correlated with histopathological characteristics and patient outcomes after a mean follow-up of 55 months. RESULTS: We observed a relationship between E-cadherin expression and disease recurrence. Disease recurrence occurred in 87.5% of patients with strong E-cadherin expression. Only 50.0% of patients with moderate expression and 0% of patients with weak or no expression of E-cadherin had disease recurrence (p = 0.014). There was also a difference in disease-free survival. Patients with strong E-cadherin expression had a mean disease-free survival rate of 49.1 months, compared to 83.9 months for patients with moderate expression (p = 0.011). Additionally, an absence of α-catenin expression was associated with tumors that were larger than 3 cm (p = 0.003). CONCLUSIONS: We demonstrated for the first time that immune expression of E-cadherin is related to tumor recurrence and disease-free survival rates, and the absence of α-catenin expression is related to tumor size in upper tract urothelial carcinomas.

Ewing's Sarcoma of the Lesser Sac Masquerading as a Pancreatic Tumor

Extraosseous Ewing's sarcoma/primitive neuroectodermal tumor (ES/PNET) is an uncommon, aggressive, and malignant tumor with a poor patient outcome. Its occurrence in the lesser sac is a rare event and to the best of our knowledge, has not been previously described. The present case was clinically and radiologically misdiagnosed as a pancreatic tumor/gastrointestinal stromal tumor. Histopathology revealed a tumor with "small round cells" that were positive for CD99, confirming the diagnosis of ES/PNET. This report highlights the importance of considering Ewing's sarcoma in the differential diagnosis of intraabdominal, extraintestinal masses.

Ewing's sarcoma of the mandible in a young child

Ewing's sarcoma (ES) is a malignancy primarily affecting bone tissue that is commonly diagnosed in adolescents and young adults. Its occurrence in the head and neck region is unusual and generally involves the mandible and maxilla. An extensive review of the literature shows only few cases of the oral ES in patients under the age of 5. This paper reports a rare case of ES of the mandible in a 4-year-old girl, which had been previously misdiagnosed and treated as a dental abscess. In the clinical examination, a hard immobile expansive mass of 5 cm in diameter was observed on the left side of the mandible. Radiographic examination revealed a radiolucent lesion with ill-defined borders and wide vestibular bone plate destruction. Microscopically, the tumor was composed by monotonous small round cells that exhibited immunoreactivity for CD99, vimentin and pancytokeratin. The patient was subjected to multiagent chemotherapy with ifosfamide, carboplatin, etoposide, vincristine, cyclophosfamide and doxorrubycin (VAC/ICE regimen). However, after the first chemotherapeutic cycle, the patient died due to disseminated infection. This case elucidates the importance of professional knowledge of the relevant aspects of malignant lesions such as ES.

O sarcoma de Ewing é um tumor maligno primário do osso, comumente diagnosticado em adolescentes e adultos jovens. Sua ocorrência na região de cabeça e pescoço não é usual, e geralmente ocorre em maxila ou mandíbula. Após revisão extensiva da literatura, poucos casos foram identificados acometendo pacientes com menos de 5 anos de idade. Nós relatamos um caso raro de SE em uma criança de 4 anos de idade que foi previamente diagnosticada e tratada como abscesso dentoalveolar. Ao exame clínico, uma massa expansiva endurecida e imóvel de 5 cm de diâmetro foi observada no lado esquerdo da mandíbula. O exame radiográfico mostrou lesão radiolúcida, com bordas mal definidas e ampla destruição da tábua óssea vestibular. Microscopicamente, o tumor era composto por células pequenas e arredondadas que exibiam imunorreatividade para CD99, vimentina e pancitoqueratina. O paciente foi submetido à quimioterapia com ifosfamida, carboplatina, e etoposide além de vincristina, ciclofosfamida e doxorrubicina (regime VAC/IE). Entretanto, após o primeiro ciclo da quimioterapia, o paciente foi a óbito por infecção disseminada. Este caso salienta a importância do conhecimento profissional no diagnóstico de tumores malígnas tais como o SE.

Inibição in vivo das óxido nítrico sintases: efeitos sobre a expressão de moléculas de adesão e secreção de mediadores inflamatórios / In vivo inhibition of nitric oxide synthases: effects on expressions of adhesion molecules and secretion of inflammatory mediators

Resultados preliminares do nosso grupo de pesquisa demonstraram que a inibição da síntese de NO por período de tempo prolongado reduz o recrutamento de leucócitos para focos inflamatórios, dependente, pelo menos em parte, da inibição da interação leucócito-endotélio e da expressão de L-selectina. 0 presente trabalho visou complementar os estudos sobre os mecanismos envolvidos nesta ação antiinflamatória. Para tanto, ratos Wistar machos (180 a 220g) receberam L-NAME (20mg/Kg; v.o.; 14 dias) ou água pela mesma via e período de tempo. Foi avaliada a atividade das enzimas óxido nítrico sintases (NOS) no tecido cerebral por radioimunoensaio; o recrutamento leucocitário para cavidade peritoneal induzido pelo LPS (5mg/kg, 4 horas); as expressões de moléculas de adesão em leucócitos do sangue circulante, no músculo cremaster e nos sinusóides hepáticos por ensaios de citometria de fluxo ou imunohistoquímica; as expressões gênicas de moleculas de adesão, quantificadas por PCR e a secreção/produção de mediadores inflamatórios em leucócitos por ensaios imunoenzimáticos, reacao de Griess ou quimiluminescência. Os resultados obtidos mostraram que: 1) o tratamento com L-NAME reduziu em torno de 90% a atividade das NOS dependentes de Ca+2 em condições basais ou após estimulação in vivo com LPS; 2) as concentrações de NO no plasma e no peritônio inflamado estavam reduzidos em animais tratados com L-NAME (30% vs. controles); 3) a migração de leucócitos polimorfonucleares (PMN) para o peritônio inflamado estava reduzida em animais tratados com L-NAME (40% vs. controles); 4) as expressões de L-selectina e PECAM-1 em leucócitos circulantes; de PECAM-1 no endotélio do músculo cremaster e de VAP-1 no endotélio dos sinusóides hepáticos e músculo cremaster de animais tratados com L-NAME estavam reduzidas; 5) a redução na expressão de L-selectina foi dependente de inibição de sua síntese; 6) a concentração de IL-10 estava major no soro de animais tratados com L-NAME em relação aos controles; 7) a maior concentração de IL-10 circulante pode refletir a produção desta citocina por leucócitos na circulação, uma vez que a concentração de IL-10 também estava maior no sobrenadante de leucócitos circulantes de animais tratados com L-NAME; 8) concentrações reduzidas de IL-1ß e LTB4 foram detectadas nos sobrenadantes de neutrófilos obtidos de animais tratados com L-NAME; 9) macrófagos obtidos de animais tratados com L-NAME produziram maiores concentrações de IL-1ß, TNF-α e IL-6, e menores concentrações de IL-10 na ausência de estimulação; na vigência estimulação in vitro com LPS os macrófagos de animais tratados com L-NAME produziram menores concentrações de NO. Em conjunto, os resultados obtidos neste trabalho mostram que o tratamento com L-NAME por período prolongado de tempo inibe a atividade das NOS dependentes de Ca+2 e nesta condição reduz as concentrações de NO circulante e no foco da inflamação, além de inibir a migração de leucócitos para o foco inflamatório, confirmando propriedades pro-inflamatórias do NO. Os mecanismos envolvidos na inibição da migração celular parecem compreender a modulação da expressão e/ou síntese das moléculas de adesão constitutivas expressas em leucócitos e no endotélio, além da modulação da secreção de mediadores pró ou antiinflamatórios em leucócitos circulantes e neutrófilos. Por outro lado, os efeitos do tratamento com L-NAME sobre a secreção de mediadores químicos por macrófagos induzem a secreção destes e corroboram a dualidade dos efeitos do NO no processo de recrutamento celular

Our previous results have demonstrated that in vivo chronic inhibition of nitric oxide synthesis reduces leukocyte recruitment into inflammatory focus, dependent, at least in part, on impaired leukocyte-endothelial interactions and expression of L-selectin. This study aimed to clarify the mechanisms involved in the reduced leukocyte migration observed in L-NAME-treated rats. For this purpose, male Wistar rats (180-220g) were treated with L-NAME (20 mg/kg, oral route, 14 days, dissolved in drinking water); controls animals received water by the same route and period of time. The effectiveness of L-NAME treatment was investigated by examining the activity of nitric oxide synthases (NOS) in the brain tissue using radioimmunoassay. In addition, effects of L-NAME treatment were evaluated in LPS-induced leukocyte recruitment into peritoneal cavity (5mg/kg, 4 hours); expression of adhesion molecules was determined in circulating leukocytes, cremaster muscle and liver sinusoids by flow cytometry and immunohistochemistry assay; gene expressions of adhesion molecules were quantified by PCR and leukocyte secretion of inflammatory mediators was measured by immunoenzimatics assays, Griess reaction or chemiluminescence. Our results show that: 1) L-NAME treatment reduced, around 90%, Ca+2 - dependent NOS activity in the presence or not of in vivo inflammation; 2) concentrations of NO in the plasma and into inflamed peritoneum were reduced in L-NAME-treated animals (30% vs. control animals); 3) migration of PMN leukocytes into inflammed peritoneum was impaired in L-NAME-treated rats (40% vs. control animals); 4) expressions of L-selectin and PECAM-1 in circulating leukocytes, PECAM-1 in endothelium from cremaster muscle and VAP-1 in endothelium from liver sinusoids and cremaster muscle were reduced in L-NAME-treated rats; 5) decrease in L-selectin expression was dependent on inhibition of its synthesis; 6) concentrations of IL-10 was higher in serum from L-NAME-treated rats in comparison to control rats; 7) these higher concentrations of circulating IL-10 can reflect the production of this cytokine by leukocytes from circulation, as IL-10 levels was greater in the supernatant of circulating leukocytes obtained from L-NAME-treated animals; 8) L-NAME treatment disturbed neutrophils ability to secrete IL-1ß e LTB4, since these concentrations were lower in the supernatants of neutrophils from L-NAME-treated animals; 9) in absence of stimulation, macrophages obtained from L-NAME-treated rats produced higher concentrations of IL-1ß, TNF-α e IL-6, and lower concentrations of IL-10, whereas in presence of in vitro LPS these cells produced lower concentrations of NO. Taken together, our results show that L-NAME treatment administrated for a prolonged period of time inhibits Ca+2 -dependent NOS activity, and in this condition, reduces concentrations of NO in plasma and into inflammatory focus and decreases leukocyte migration to the inflammatory focus thus confirming pro-inflammatory properties of NO. The mechanisms involved in impaired cellular migration seem to involve the modulation of expression and/or synthesis of constitutive adhesion molecules in leukocytes and endothelium, and to interfere in the secretion of pro or anti inflammatory mediators. On the other hand, actions of NO in secreation of chemical mediators by macrophages induces the production of inflammatory mediators and support the duality of NO in the cellular recruitment process

Comparison of Sporadic Sclerotic Fibroma and Solitary Fibrous Tumor in the Oral Cavity

Sporadic sclerotic fibroma (SF) and solitary fibrous tumor (SFT) arising in the oral cavity are very rare. In this report, we describe two cases of oral pathology, one involving SF and the other involving SFT. Both cases presented with well- circumscribed, firm nodules with similar gross findings. However, the histologic findings of the SF and SFT showed rather distinct features. The SF was composed of hyalinized sclerotic collagen bundles arranged in a whorled pattern, whereas the SFT was formed by spindles cells arranged in hypo- and hypercellular areas. The immunohistochemical findings were similar in both cases; there was positivity for vimentin, CD34, and CD99, but bcl-2 positivity was only seen in the SFT. Although their histopathologies are similar, SF and SFT should be considered in the differential diagnosis of soft tissue tumors in the oral cavity.

Extraskeletal Ewing's Sarcoma of the Hard Palate

Extraskeletal Ewing's sarcoma (EES) is a rare soft tissue tumor morphologically indistinguishable from the more common Ewing's sarcoma of bone. We report a case of EES arising in the hard palate of 34-yr-old male patient. Microscopically, the monotonous small round cells without neuronal differentiation showed membranous positive immunoreactivity for MIC2/CD99 and vimentin. Ultrastructurally, the tumor cells showed a few intracytoplasmic organelles without evidence of neurosecretory granules or neurofilaments. The EWS-FLI1 chimeric gene was identified using the nested reverse transcriptase-polymerase chain reaction.

Estudo comparativo da expressão da molécula de adesão intercelular-1, da interleucina-4 e interferon-gama na dermatite de contato das mãos alérgica e irritativa / Comparative study of intercellular-1 adesion's molecule expression, interleukina-4 and interferon gamma in allergic and irritative contact dermatitis hands

Trinta pacientes com dermatite de contato das mãos, quinze com a forma alérgica e quinze com a irritativa, diagnosticados clinicamente e por meio do teste de contato, foram submetidos a avaliação quantitativa das moléculas de adesão intercelular-1 e das citocinas IL-4 e interferon-gama a fim de se obter uma diferenciação mais precisa e elucidação fisiopatológica entre os dois tipos de dermatite. Para isso foram colhidas biopsias de lesões ativas com posterior análise imunohistoquímica em tecido criopreservado, seguida da quantificação histomorfométrica na derme / Thirty patients with contact dermatitis of the hands, fifteen with the allergic form and fifteen with the irritative form, diagnosised clinically and by contact test, were submitted of the quantitative evaluation of molecules of intercellular-1 adhesion and of the cytokines IL-4 and interferon-gamma to get the most necessary differentiation and physiopathologic briefing between the two types of dermatitis. For that study, they had been harvested biopsy of active injuries with posterior immunohistochemical analysis in fabric preserved in ice, followed of the hystomorphometric quantification in dermal...

Tissue expression of platelet endothelial cell adhesion molecule-1 at pre and postnatal murine development

Endothelial cells, at the cell-cell borders, express PECAM-1, and have been implicated in vascular functions. The monoclonal antibody MEC 13.3 recognizers PECAM-1 molecule from mouse vessels and allows to analyse the ontogeny of mouse endothelium. At the present, little is known about the molecular basis of differentiation pathways of endothelial cells, that enables its morphological heterogeneity. The purpose of this study was to analyze the pattern of PECAM-1 expression, employing monoclonal antibody MEC 13.3, in cellular suspensions obtained from different mouse organs at pre and postnatal stages. Fluorescence activated cell sorter analysis showed a different profile of the glycoprotein expression in a cell population with size and granularity selected by 1G11 endothelial cell line. The expression differs from prenatal to postnatal developmental stages in a given organ, and among the organs studied. Another cell population, with a size and granularity higher than 1G11 endothelial cell line, coexists in cellular suspensions obtained from liver, gut and brain. These cells could be related to those detected by means of immunoenzyme methods which showed a non-differentiated morphology. The different PECAM-1 pattern expression could reflect potential organ-specific differentiation pathways during development and according to organs environment. The existence of another cell population with a size and granularity higher than 1G11 endothelial cell line required a phenotypic characterization.

Immunoreactivity of CD99 in Stomach Cancer

CD99 is characteristically expressed in Ewing's sarcoma/primitive neuroectodermal tumor. Recently its immunoreactivity has also been reported in other tumors. However, the significance of CD99 isoforms expressed in these tumors has not been elucidated. In this study, we evaluated the expression of CD99 isoforms and its relationship with histopathologic parameters in gastric adenocarcinomas. Paraffin sections of 46 gastric adenocarcinomas were stained with an anti-CD99 monoclonal antibody, YG32. Twelve (26.1%) cases of 46 gastric adenocarcinomas showed immunoreactivity to YG32. The CD99 expression was also seen both in non-neoplastic foveolar epithelial cells and infiltrating lymphocytes. In addition, Western blot and RT-PCR analyses revealed that the type I is the predominant isoform of CD99 in non-neoplastic and neoplastic gastric tissues. The CD99 expression was usually seen in the intestinal type adenocarcinoma, while rarely in the diffuse type. The CD99 immunoreactivity decreased in MMP-2-overexpressing adenocarcinomas (p=0.028). Our results suggest that the type I is the major isoform of CD99 expressed in non-neoplastic gastric mucosa and gastric adenocarcinomas and its downregulation in gastric adenocarcinoma may be associated with cellular dedifferentiation and/or MMP-2 overexpression.

A study on the immunocytochemical localization of neurofascin in rat sciatic nerve

We examined the localization of neurofascin (NF) in the sciatic nerve of rat. In the myelinated fibers, neurofascin localizes strongly in the nodal axolemma except the small central cleft and also expresses in the paranodes, and weakly in the Schmidt-Lanterman incisures. In the paranodes, NF localizes around the axolemma and it expresses in the apposing membrane of paranodal loops. Axoplasm, compact myelin and cytoplasm of Schwann cell do not express NF at all. In the Schmidt-Lanterman incisures, NF is expressed weakly along the Schwann cell membrane. We propose that neurofascin may be a plasmalemmal integral protein of Schwann cell in the paranode and plays some important roles for the maintenance of axo-glial junctions at the paranode. It may also have some roles for maintaining the structure of Schmidt-Lanterman incisure and have some relations with proteins localizing in the node.

Flocculation of fine fluorite particles with corynebacterium xerosis

The treatment of fine particles dispersed in liquids is common in several industries and especially important in mineral processing. The efficiency of settling operations can be susbstantially increased by flocculation. The aim of this work was to study the flocculation of fine fluorite particles by the bacterium Corynebacterium xerosis. Flocculation tests, microelectrophoresis measurements and optical microscopy were used to evaluate flocculation. The results showed that C. xerosis cells adhere to the fluorite surfaces promoting the aggregation of the particles. High quality flocs can be obtained rapidly at ph 7.0 using a cell concentration of 40 mg/l, considerably lower than previously reported in the literature. The results are discussed with reference to the surface characteristics of the mineral and of the microorganism.

High CD99 expression in memory T and B cells in reactive lymph nodes

We investigated the expression of CD99 in 35 hyperplastic perigastric lymph nodes, which were resected for gastric carcinoma or chronic peptic ulcer. Essentially, all lymphocytes in lymph nodes expressed CD99, but there were two populations with respect to the intensity of CD99 expression--CD99high and CD99low cells. We showed CD99high cells were distributed in paracortical and medullary cords by immunohistochemical study while germinal center cells were CD99low. Using three-color flow cytometric analysis with CD3, CD4, CD8, CD19, CD23, CD45RA, CD45RO, CD69, CD138, IgM, IgD, and IgG, most of CD99high cells were shown to be activated/memory T cells. CD4+CD45RO+ T cells were the subset revealing the highest intensity of CD99 expression while CD4+CD45RA+ T cells were CD99low. Among B cells, IgG+ B cells revealed a higher level of CD99 molecules than IgM+ B cells. These results suggest that CD99 is one of activation-related molecules which are upregulated in recently activated lymphocytes.

Virulence factors in fungi of systemic mycoses

Fungos patogênicos causadores de micoses sistemicas possuem vários fatores que permitem seu crescimento nas condiçöes adversas oferecidas pelo hospedeiro, propiciando o estabelecimento da relaçäo parasitária e contribuindo no processo de doença. Esses fatores säo conhecidos como fatores de virulencia auxiliando no desenvolvimento da infecçäo e interferindo com a patogenese das micoses. O presente trabalho avalia os fatores de virulencia em fungos patogenicos como Blastomyces dermatitis, Coccidioides immitis, Cryptococcus neoformans, Histoplasma capsulatum e Paracoccidioides brasiliensis, em relaçäo a termotolerancia, dimorfismo, componentes da parede celular ou capsula, bem como a producao de enzimas...

Proteoglycan's activation by adhesion molecules and L metalloproteases in rheumathoid arthritis and ostheoarthritis

Two groups of patients were studied, both in accordance with ACR criteria. First group (41 cases) suffering R.A.. Second group (36 cases) suffering O.A. In both pathologies MMPs, ICAM and VCAM from synovial fluid and plasma were studied. Measurements were made with ELISA-sandwich in a Metrolab spectrophotometer at 410 mm for MMPs, and 491 nm for ICAM and VCAM. As control, samples of patients with noninflammatory muscle skeletal disorders or traumatic arthritis and healthy witness were used. Synovial concentration of MMPs in R.A. was 1402+76 ng/ml, a higher significant value (p<0.0001) compared with ostheoarthritis: 353+23 ng/ml. In the witness plasma, MMPs were not detected. Plasmatic and synovial levels of the adhesion molecules present different values in both pathologies and between them. Synovial ICAM level in R.A. (280+9.8 ng/ml) is significantly higher than in O.A. (163+10 ng/ml) (p<0.001), but lower than plasmatic ones (370+35 ng/ml) (p,0.001). All these values are significantly higher than the normal plasma (121+6.5 ng/ml) (p<0.005, and p<0.0001, respectively) VCAM increase regarding basal values (140+5.6 ng/ml) (p<0.001) and in a similar proportion for both pathologies (R.A.: 186+9.3 ng/ml and O.A.: 207+14.3 ng/ml). Their plasmatic levels were higher (270+45 and 320+38 ng/ml) (p<0.001) but without significative difference between them. There is correlation among MMPs, ICAM and VCAM variations. The variability can be explained by concomitance several evolutive steps. Each pathology shows a different grade of cellularity, inverted predominance in the relation TIMPs/collagenase and different generator mechanisms of MMPs. Our findings reinforce the importance as diagnostic guide of adhesion molecules dosage, and possible therapeutic use of MMPs inhibitors and ICAM ou VCAM antagonists en R.A. and related pathologies.

Determinación de ICAM-I soluble en suero de pacientes con enfermedad de Hansen / Soluble ICAM-I determination in patients serum with hansen's disease

En este estudio se detectó la forma soluble ICAM-1 (ICAM-1s) en el suero de pacientes con lepra tuberculoide (TT,n=7), lepra indeterminada (l1,n=7), lepra bordeline (BB,n=8), lepra lepromatosa (LL,n=6) e individuos clínicamente curados de lepra lepromatosa (LL curados,n=6). Como controles se analizaron sueros de individuos de áreas no endémicas (n=5). La metodología utilizada para las determinaciones consistió en un enzayo inmunoenzimático (ELISA) de captura, específico para ICAM-I. Los resultados mostraron niveles significativamente elevados de ICAM-1s en pacientes con LL (790+/-152ng/ml) y TT (556+/-112ng/ml). Los niveles de ICAM-1s en suero de pacientes con L1 (270+/-32ng/ml) y BB (269+/-46ng/ml) fueron muy similares a los individuos control sano (250/-11ng/ml). Una observación importante fué la significativa reducción de los niveles de ICAM-1s en los pacientes con LL luego del tratamiento con poliquimioterápia (LL curados, 412+/-105ng/ml). Aún se desconoce el papel inmunorregulador de las móleculas de adhesión circulantes en los procesos de inflamación, sin embargo se ha sugerido que niveles fisiológicos están asociados con activación leucocitaria y niveles altos con daño tisular. En los pacientes con LL los altos de ICAM-1s podrían competir por los ligandos de ICAM-1 (mólecula LFA-1) en los linfocitos T, causando así parte de la característica energía selectiva. El moderado aumento de ICAM-1s en los pacientes con TT podría estar asociado con el estado inflamatorio activo de estos pacientes. Una conclusión similar podría esbozarse en la marcada reducción de ICAM-1s observada en los LL curados, donde se observa un retorno a niveles normales.