In Situ Cytokine Expression and Morphometric Evaluation of Total Collagen and Collagens Type I and Type III in Keloid Scars.

Keloids are characterized by excessive collagen deposition and growth beyond the edges of the initial injury, and cytokines may be related to their formation. The objective of this study was to evaluate the collagen fibers, analyze in situ expression of cytokines in keloid lesions, and compare to the control group. Results showed that there was a predominance of women and nonwhite and direct black ancestry. Keloid showed a significant increase in total and type III collagen. Significantly, the expression of mRNA for TGF-ß in keloid was increased, the expressions of IFN-γ, IFN-γR1, and IL-10 were lower, and IFN-γR1 and TNF-α had no statistical difference. Correlations between collagen type III and TGF-ß mRNA expression were positive and significant, IFN-γ, IFN-γR1, and IL-10 were negative and significant, and TNF-α showed no statistical difference. We conclude that there was a significant increase of total collagen in keloid and predominance of collagen type III compared to the controls, showing keloid as an immature lesion. There is a significant increase in TGF-ß mRNA in keloid lesions, and a significant decrease in IFN-γ and IL-10, suggesting that these cytokines are related to keloid lesions.

Stainless steel weld metal enhanced with carbon nanotubes.

This paper aims to establish the most indicated route to manufacture a nanostructured powder composed of 5 wt% Multi-walled Carbon Nanotubes and 304LSS powder. Four specimens were prepared using Mechanical Alloying and Chemical Treatment (CT) with Hydrogen Peroxide ([Formula: see text]) as the main processes. A thermal treatment post-processing was used in half of the samples to remove the remaining amorphous carbon and to evaluate its effects. Regarding the powder analysis, attachment, amorphous carbon degree, crystallinity, and doping of the CNT throughout the metal matrix were investigated. The nanostructured powders were then inserted as a core in a 304LSS tubular rod to perform the arc welding process. The CT route eliminated the amorphous carbon and generated more refiner grains, which provided a cross-section hardness gain of more than 40% regarding the 304LSS joint. In summary, the CT route, combined with the GTAW process, provided a new method for nanocomposite manufacturing by combining shorter preparation steps, obtaining an improvement in the microstructural and hardness performance.

Mast cells, TGF-beta1 and alpha-SMA expression in IgA nephropathy.

IgA nephropathy (IgAN) is a kidney disease with a varying renal prognosis. Recently, many studies have demonstrated that renal alpha-smooth muscle actin (alpha-SMA) and transforming growth factor (TGF-beta1) expression, as well interstitial mast cell infiltrates could represent a prognostic marker in several renal diseases. The aim of our study was to analyze the prognostic value of mast cell, TGF-beta1 and alpha-SMA expression in IgAN. A survey of the medical records and renal biopsy reports of 62 patients with a diagnosis of IgAN followed-up from 1987 to 2003 was performed. The mean follow-up time was 74.7 +/- 50.0 months. The immunohistochemical studies were performed using a monoclonal antibody anti-human mast cell tryptase, a polyclonal antibody anti-human TGF-beta1, and a monoclonal antibody anti-human alpha-SMA. An unfavorable clinical course of IgAN was related to interstitial mast cell infiltrates and alpha-SMA expression in the tubulointerstitial area. Expression of glomerular TGF-beta1 and alpha-SMA, and interstitial TGF-beta1 is not correlated with clinical course in IgAN. In conclusion, the increased number of mast cells and higher alpha-SMA expression in the tubulointerstitial area may be predictive factors for the poor prognosis of patients with IgAN.

A role for HO-1 in renal function impairment in animals subjected to ischemic and reperfusion injury and treated with immunosuppressive drugs.

INTRODUCTION: Ischemia/reperfusion injury (IRI) represents the single major antigen-independent factor implicated in pathogenesis of chronic graft dysfunction. Tacrolimus is a calcineurin inhibitor, which has been suggested to be helpful in cyclosporine-related chronic toxicity. Rapamycin has antiproliferative properties that may impair renal regeneration after IRI. Therefore, immunosuppressive drugs might impair renal graft outcome in those organs suffering IRI. MATERIAL AND METHODS: C57B1/6 male mice subjected to 45 minutes of renal pedicle ligation were reperfused for 24 hours. Mice were treated with rapamycin, cyclosporine, or tacrolimus. Blood and renal tissue samples were collected at 24 hours after IRI. Urea levels were measured. Heme Oxygenase 1 (HO-1) gene transcript was amplified by a real-time polymerase chain reaction technique. RESULTS: Animals treated with cyclosporine and subjected to IRI showed impaired renal function that peaked at 24 hours. Additional pretreatment with rapamycin produced even more impairment of renal function, when compared with controls. However, tacrolimus pretreatment was associated with a better renal outcome. HO-1 expression was upregulated after IRI by 2.6 arbitrary units at 24 hours. Rapamycin showed worse impairment of renal function. CONCLUSION: Tacrolimus was not associated with worsening renal function when compared with animals just subjected to IRI. Upregulation of HO-1 may be an attractive approach to limit graft injury.

The effects of rapamycin in the progression of renal fibrosis.

UNLABELLED: Renal fibrosis is a hallmark of end-stage renal diseases and of chronic allograft nephropathy (CAN). Rapamycin, besides its action through blockade of lymphocyte proliferation, also has antiproliferative, antiviral, and antitumor actions. Its use in clinical in patients with CAN has recently been advocated. OBJECTIVES: Our goal was to evaluate the effect of rapamycin in an established model of renal fibrosis, unilateral ureteral obstruction. MATERIALS AND METHODS: C57BL/6 mice were divided into two groups, treated or not with daily doses of rapamycin (0.2 mg/kg) beginning on day-1. The obstruction was performed as day 0. Blood and kidney tissues were collected at 1, 4, 7, and 14 days after the surgery to quantify bone morphogenic protein (BMP)-7 and transforming growth factor (TGF)-beta mRNA by real time PCR. RESULTS: Daily treatment with rapamycin caused a significant reduction in serum creatinine at day 1 (0.57 +/- 0.03 vs 0.95 +/- 0.15 mg/dL, P = .002) and at day 14 (0.56 +/- 0.04 vs 0.73 +/- 0.07 mg/dL, P = .040). This profile was corroborated by histological morphometric analyses showing less fibrosis at day 14. However, rapamycin surprisingly induced an upregulation of TGF-beta at day 4 (3.05 +/- 0.46 vs 1.85 +/- 0.41, P = .006) and at day 7 (6.33 +/- 0.55 vs 4.97 +/- 0.38, P = .024) with a reduced expression by day 14 (4.03 +/- 1.07 vs 7.89 +/- 0.83, P < .001). Surprisingly, rapamycin also promoted an increment in BMP-7, completely reversing the ratio of TGF-beta to BMP-7, allowing a more protective phenotype. CONCLUSION: Rapamycin slightly ameliorated the renal dysfunction and, at later time points, induced less fibrosis and less decrease in the TGF-beta to BMP-7 ratio.

Quantitative analysis of fibrosis and mast cells in the tongue of chronic chagasic patients: autopsy study.

OBJECTIVE: Recently, an orally transmitted outbreak of Chagas disease was reported in Santa Catarina, Brazil, after ingestion of sugar cane juice (garapa). This disease is caused by Trypanosoma cruzi, a parasite that stimulates the development of chronic inflammatory response, characterized by fibrous connective tissue neoformation (fibrosis). As the density of tissue mast cells (MC) may be an index of fibroblast proliferation and development of local fibrosis, the purpose of this autopsy study was to quantify the fibrosis rate and the number of MC in the tongues of chronic chagasic (CC) patients, compared with a non-chagasic (NC) control group. METHODOLOGY: Twenty-four evaluations, with a quantitative assessment of fibrosis percentage and MC density were performed. RESULTS: The percentage of fibrosis in the tongue was higher among CC than in the control group. In the CC group, a positive and significant correlation was found when the fibrosis rate was compared with the MC density. CONCLUSIONS: These morphometric findings suggest that tongue biopsy may be useful to study specific changes associated with Chagas disease. They also suggest that the systematic analysis of oral cavity, including tongue histopathology changes, could be useful in forensic pathology of the orally acquired chronic Chagas disease.

Dual response of animals with chronic cyclosporine nephrotoxicity to an acute ischemic injury.

Normal kidneys regenerate after acute injury with little development of chronic fibrosis. However, the long-term effects of an acute injury in kidneys with established chronic toxicity induced by cyclosporine (CsA) are not entirely clear. To study the consequences of an ischemia and reperfusion (IR) injury in long-term CsA-treated rats, male Wistar rats (250-300 g) were treated daily with CsA (10 mg/kg) or vehicle (olive oil 1 mL/kg) for 28 days. On day 21, ischemia was performed by clamping the renal vessel for 1 hour. Blood samples were collected on days 0 and 21 (before IR) as well as days 22 and 28. On day 28, the kidneys were collected to examine the mRNA expression of MCP-1 by real-time PCR. For renal function, serum creatinine levels were measured. Twenty-four hours after reperfusion, long-term CsA-treated animals showed better renal function compared with the control group, as demonstrated by serum creatinine levels: 2.2 +/- 0.13 mg/dL vs 2.9 +/- 0.18 mg/dL, respectively (P < .05). However, 1 week after IR, the renal function was worse among the long-term CsA-treated group than the controls: 1.16 +/- 0.08 mg/dL vs 0.8 +/- 0.09 mg/dL, respectively (P < .05). Interestingly, CsA treatment was associated with lower MCP-1 mRNA expression than that in the control group: mean MCP-1 mRNA expression 0.58 +/- 0.13 vs 1.02 +/- 0.12, respectively (P < .05). In conclusion, animals with chronic CsA nephrotoxicity were protected from an acute renal injury, possibly through decreased chemokine production, although at later time points, renal function was clearly impaired, probably by the acceleration of vasculopathy caused by nephrotoxicity.

Alpha-SM actin expression as prognostic indicator in IgA nephropathy (Berger's disease).

Recent studies have demonstrated that alpha-Smooth Muscle actin expression in glomerular and tubulointerstitial compartments of renal tissue could represent a prognostic marker in several renal diseases. Our objective was to identify the prognostic value of alpha-SM actin actin expression on the evolution of renal damage in Primary IgA nephropathy (Berger's disease). 43 patients followed up from 1988 to 1999 at the University Hospital, Faculty of Medicine of Ribeirão Preto, University of Sao Paulo, Brazil, was studied. Clinical-laboratory data were obtained from the medical records of the patients using a protocol containing name, race, gender, origin, profession, age at clinical presentation of the disease and personal and family history. The parameters assessed in the approach to IgA nephropathy were serum creatinine, creatinine clearance, serum albumin, total serum protein, 24 hours proteinuria, glycaemia, serum sodium, potassium, calcium and phosphorus ions, analysis of urinary sediment, serum complement profile, blood count, and renal biopsy. Morphological evaluation was performed by renal biopsy using common light and immunofluorescence microscopy. Immunohistochemical studies were performed using a murine monoclonal antibody to alpha-SM actin. Our data showed that alpha-SM actin expression in the glomerular and tubulointerstitial compartments are not correlated with unfavorable clinical course of primary IgA nephropathy.

Physical training induces expression of TGF-beta in kidney of pregnant hypertensive rats.

AIM: This study aimed to determine the morphological renal impairment in pregnant rats spontaneously hypertensive (SHR) submitted to swimming when compared with those who did not perform the activity, and to analyze the relationship of expression of cytokines in inflammatory fibrotic and protrained and sedentary animals. METHODS: SHRs and their respective control normotensive rats (WKY) were submitted or not to a swimming protocol during 9 weeks, resulting in four pregnant experimental groups: sedentary hypertensive (HS), trained hypertensive (HT), sedentary normotensive (NS), and trained normotensive (NT). RESULTS: Pregnant untrained hypertensive rats presented higher resting mean blood pressure (MAP) compared with both sedentary and trained normotensive groups (P<0.05). We can observe too, that the exercise training did not change the heart rate (HR) in both hypertensive and normotensive groups (P=0.127). The HT rats showed lower area of mesangial matrix (MM) compared to NT group (P=0.018). The perceptual of fibrosis (%F) in hypertensive rats was significantly higher compared with the % F in normotensive rats (P<0.001). The rats in the HT group showed higher expression of TGF-b (P<0.001) and of IL-10 (P<0.001) when compared with the other groups. CONCLUSION: The main conclusion is that in SHR rats it is shown a greater expression of TGF-beta, resulting in increased fibrosis in renal parenchyma due to the increased number of inflammatory cells that secrete this cytokine, and thus the practice of swimming can attenuate inflammatory processes, and mitigate the blood pressure of these animals.

Correlation between adrenal central vein parasitism and heart fibrosis in chronic chagasic myocarditis.

The correlation between Trypanosoma cruzi parasitism of the adrenal central vein (ACV) wall and fibrous connective tissue neoformation in the left ventricular myocardium (LVM) of patients with chronic Chagas' disease who were autopsied was evaluated using the following procedures: 1) a comparison of the incidence of fibrosis in the LVM among 18 chagasic patients with ACV parasitism and 18 individuals without phleboparasitism; 2) a determination of fibrosis intensity in the LVM in 12 cases with ACV parasites and in 12 cases without phleboparasitism, matched with respect to age, sex, race, and anatomoclinical form of the disease (indeterminant, cardiac, and digestive forms); and 3) in the cases with ACV parasitism, a calculation of Pearson's correlation coefficient between T. cruzi nests in the vessel and the intensity of fibrous connective tissue neoformation in the LVM. Among chagasic individuals with adrenal phleboparasitism, there was an increased incidence and intensity of fibrous connective tissue neoformation in the LVM, both highly significant, compared with patients without adrenal phleboparasitism. Furthermore, there was a positive, though nonsignificant, correlation (r = +0.19) between the density of nests in the ACV and the intensity of myocardial fibrosis. These results are consistent with previous data showing a higher intensity of the leukocyte exudate in the LVM and increased heart weight in individuals with T. cruzi nests in the ACV, suggesting a role of parasitism at that site in terms of the development of chronic chagasic cardiopathy.

Possible role of an adrenal parasite reservoir in the pathogenesis of chronic Trypanosoma cruzi myocarditis.

The possible contribution of parasitism of the central vein of the adrenal gland (CVAG) to chronic Trypanosoma cruzi myocarditis was assayed by comparing the occurrence of nests of amastigotes in the left ventricular myocardium (LVM) and determining the number and extent of areas of focal leucocyte exudate (FLE) in Chagas disease patients with and without CVAG parasitism. The frequency of occurrence of T. cruzi nests in the LVM, as well as the FLE number and area, were greater among patients with CVAG parasitism. We therefore suggest that CVAG parasitism plays a role in the onset and intensity of chronic T. cruzi myocarditis.

[Renal anatomopathological changes in patients with acquired immunodeficiency deficiency syndrome]. / Alterações anatomopatológicas renais em indivíduos com a síndrome da imunodeficiência adquirida.

Renal anatomopathological lesions were studied among 119 AIDS patients from Faculdade de Medicina do Triângulo Mineiro's University Hospital (Uberaba, MG, Brazil). From formalin-fixed blocks, slides were obtained and studied by light microscopy. Of 119 patients, 67 presented tubulointerstitial nephritis (TIN), 18 inespecific, 2 xantogranulomatous and infections agents were found in 48 as follows: mycosis in 28 (16 Cryptococcus sp; 9 Histoplasma sp, 1 Candida sp e 2 Paracoccidioides brasiliensis); bacteria in 18 (9 Mycobacterium sp), virus in 6 (Cytomegalovirus). Acute tubular necrosis was found in 43 cases (36.1%). Other diagnosis were: nefrocalcinosis (15.1%), arteriolar hyalinosis (22.7%), two cases of focal segmental glomerulosclerosis (1.7%) and one case of diffuse mesangial hyperplasia (0.8%). We conclude that the renal involvement in patients with AIDS, presents a wide spectrum of pathologies, secondary to complications related to opportunistic infections, therapeutic and diagnostic management, and the nephropathies associated to HIV.

[Human accidents with stings from Africanized bees]. / Acidente humano por picadas de abelhas africanizadas.

It is report five cases of stings of africanized bees (AB). Four of them were victims of multiple stings, varying from 25 to 60, that presented severe headache, nauseas and vomits, related to the clinic effect of the toxicity of the venom, with good evolution. The other case was victim of a single sting of AB and died after probable anaphylaxis.

[Clinical and laboratory renal alterations in patients with acquired immunodeficiency syndrome in relation to anatomopathological findings]. / Alterações renais clínico-laboratoriais em pacientes com a síndrome da imunodeficiência adquirida em relação aos achados anatomopatológicos.

Clinical as well as renal anatomopathological data of 119 patients with acquired immunodeficiency syndrome from the Hospital of the Faculty of Medicine of Triângulo Mineiro, Uberaba, MG, Brazil, were studied. Mean age was 33 +/- 10.4 years (range 18 to 67 years). White (59%) and male (80%) individuals predominated. Ninety percent of patients came from Uberaba or neighborhood. There were 59 (50%) heterosexual, 25 (21%) homosexual, 17 (14%) bisexual and 18 (15%) unknown. Forty three percent were drug abusers. Fourteen (12.2%) of patients had serum creatinine higher than 3mg/dl, ten days before death, 64% of them showed acute tubular necrosis. Seventy one (61%) patients presented hyponatremia and 8 (7%) had hypernatremia. Hypokalemia was detected in 28 (24%) and hyperkalemia in 8 (7%) patients. Twenty four hours urinary protein was detected in 12 patients, 7 of them presenting more than 1g/24h. Two patients showed segmentar focal glomerulosclerosis, one of them with 24h urinary protein level of 5.5 g and serum creatinine 1.5 mg/dl and no edema. The most (56%) patients had tubulointerstitial nephritis. Clinical, laboratory and types of renal disorders in patients with acquired immunodeficiency syndrome are variable. The glomerular diseases were relatively scarce however, tubulointerstitial lesions are quite frequent, mainly interstitial nephritis and acute tubular necrosis. Not always an anatomoclinical correlation could be observed, because many times the electrolyte disorders were frequently observed only functionally.

[Prevalence of renal infarcts in autopsies of chronic Chagas disease patients]. / Prevalência de infartos renais em necropsias de chagásicos crônicos.

Renal infarction (RI) is usually secondary to arterial obstruction due to emboli originating from the heart. Chronic chagasic patients may present cardiac alterations originating from intracavitary thrombi, even without congestive heart failure (CHF). In this study RI incidence was comparatively evaluated in chronic chagasic individuals, in different anatomoclinic forms and in non chagasic individuals. There has been a review on necropsy reports of individuals aged 20 or over. In 259 necropsies, 78 (30.1%) were chagasics, and 19 of them (24.4%) developed RI, while 27 (15.0%) of the non chagasic individuals presented RI. The ages of chagasics with RI were similar to those of non chagasic individuals. A significant prevalence of RI and thrombosis among chronic chagasic individuals has been found. A significantly higher prevalence of RI among chronic chagasics having CHF (52.6%) was observed when they were compared to other forms of chronic Chagas disease and when compared to non chagasic individuals. It was concluded that RI was more frequent in chronic chagasic individuals, specially those who developed CHF, which probably played a role in the renal manifestations and systemic hemodynamic changes in those patients.

[Adrenal gland morphological alterations in the acquired immunodeficiency syndrome]. / Alterações morfológicas da glândula supra-renal na síndrome da imunodeficiência adquirida.

In the acquired immunodeficiency syndrome (AIDS), the adrenal glands are subject to opportunistic infections, neoplasm or direct cytopathic effect by HIV. It is know that the incidence and type of adrenal involvement vary according to the patient's place of origin. In this paper we evaluate adrenal involvement in fourteen patients that died from AIDS in the University Hospital of Uberaba, Brazil. The group studied was comprised of thirteen males and thirteen whites. The age was 29.9 +/- 7.8 years, and the body mass index was 19.0 +/- 4.1 kg/m2. Adrenal specimens obtained from autopsies were analyzed by light microscopy. Inflammation was found in 100% of the cases and the etiologic agent(s) was (were) identified in eight (58.1%) patients. Cytomegalovirus was identified in seven cases, Cryptococcus sp and Herpes simplex in two and Histoplasma sp in one case, these pathologic findings were similar to literature. We also found parenchymal calcification and adrenal central vein phlebitis in one case each. Injury was found in some cases without identified infections agent. This fact could be due to the direct cytopathic effect by HIV, or due to toxicity of drug therapy used during treatment of AIDS and opportunistic infections.

[Miliary tuberculosis - report of a case]. / Tuberculose miliar--relato de caso.

This is a case report of generalized miliary tuberculous infection in a 80-year old white male without the acquired immunodeficiency syndrome, whose death was caused by progressive hematogenous seeding similar to those cases of preantibiotic era. The importance of autopsy studies to uncover silent or protean infections specially in cases of cryptic or chronic hematogenous miliary tuberculosis, is emphasized.

[Comparison of parasitology of the central vein of the adrenal gland and the other tissues in chronic chagas disease]. / Comparação do parasitismo da veia central da supra-renal com o de outros tecidos em chagásicos crônicos.

By morphological and morphometric analyses of serial sections the occurrence of T. cruzi nests in the central vein and in the parenchyma of adrenal glands, in the left ventricular wall and in the inferior vena cava wall in chronic Chagasic patients was studied. Of 36 cases 50% showed parasites in the adrenal central vein wall (total 29 nests), 3.1% showed parasites in the vena caval wall (only I nest) and 16.8% we found parasites in the myocardiocytes (total 23 nests). The density of parasites measured in the nests for each 100mm2 of the tissue examined, was 0.585 for the adrenal vein, 0.001 for the vena cava and 0.01 for the myocardium. No nest was found in 269103.1mm2 of adrenal parenchyma. Although the central vein area examined was smaller, it showed the largest frequency of T. cruzi nests. Since a basic difference between these tissues is the great quantity of corticoids in the blood of the adrenal central vein, this prevalence may be because of this hormonal ambient, which with its immunosuppressor and anti-inflammatory effects could help T. cruzi survival.

[Prevalence of intestinal parasitosis in feces collected during necropsies]. / Prevalência de enteroparasitoses em fezes colhidas durante necrópsias.

With the intention to verify the frequency of intestinal parasites, we have taken the inside digestive system material from three different parts of the intestinal tract while we were doing 135 complete necropsies. The stools were kept in MIF (Formol, iodine and merthiolate mixture) being studied in slides by the direct method. We have found intestinal parasites in 40 cases (29.6%), 11 (27.5%) with two or more kinds of parasites and 29 cases (72.5%) with only one kind of parasite. The most frequent parasites were: S. stercoralis (31,9%), E. hystolytica (23.4%), hook worm (19.1%) and A. lumbricoides (17%). The results are in according with epidemiological research made in this area. Thus, they prove that the parasitological exam made in feces taken during the necroscopy provides further evidence to enrich the anatomo-pathological diagnosis.

[Contribution to the morphologic diagnosis of lung adiaspiromycosis]. / Contribuição ao diagnóstico morfológico da adiaspiromicose pulmonar.

The diagnosis of adiaspiromycosis is usually based on lung sections stained by hematoxylin-eosin, periodic acid Schiff and methenamine silver. Authors describe the fungus aspect examined by mucicarmin, picro-sirius and Congo red methods, including polarized light microscopy. In doubtful cases, these methods could contribute to histopathological diagnosis of Emmonsia parva var crescens.