C4d as a Practical Marker for Cutaneous Amyloidosis.

ABSTRACT: Cutaneous amyloidosis (CA) is defined by the accumulation of amyloid in the dermis; it might be primary or secondary. The diagnosis is based on histopathological findings with the demonstration of amyloid deposits, confirmed by Congo red stain under the polarized light. Studies on other diagnostic markers are ongoing in the literature. The aim of this study was to demonstrate the utility of C4d staining in the recognition of amyloid in CA and using it as an alternative or substitute marker for the diagnosis. In this retrospective study, 199 skin biopsies with a clinical provisional diagnosis of CA were analyzed, the Congo red stain was performed, and, in a subgroup (n = 97) with histopathological findings probably for CA, C4d immunohistochemistry was assessed. Forty-eight cases of CA were detected. Congo red birefringence was positive in all cases, whereas in 14 cases, it was faded. In these 14 cases, the diagnosis of CA was made by means of Congo red fluorescence and Thioflavin T because the histopathological findings were highly suggestive for CA. All CA cases were positive with C4d, and in 12 of the 49 inflammatory dermatoses, C4d was positive. The interpretation of C4d immunohistochemistry can be performed more easily and rapidly than Congo red evaluation. The sensitivity and specificity of C4d were 100% and 75.5%, respectively. In our experience, C4d staining was a useful method for detecting amyloid deposits in CA. Although Congo red staining is the gold standard for amyloid detection, we propose C4d immunohistochemistry as a routine screening method or hybrid transition while further investigations are completed.

Entrapment and release kinetics study of dyes from BSA microspheres forming a matrix and a reservoir system.

Two kinds of Bovine Serum Albumin (BSA)-loaded microspheres were prepared in water-organic bilayer systems using ultrasonic irradiation. The first method included an aqueous solution of BSA and water-soluble dye together, mixed with dodecane, that upon sonication formed a matrix system where the dye is concentrated in the protein shell. The other system included an aqueous solution of BSA mixed with octanol-soluble dye that, upon sonication, formed a reservoir system in which the dye filled the inner volume of the microspheres. Each of these microspheres was prepared with two different dyes and their leaching profiles into pure solvents were studied using UV-vis spectrometry. Fast leaching was observed at the beginning for both systems, which levelled-off after a certain time. For the matrix system, an equilibrium state was obtained after 100-200 hours, whereas for the reservoir system, leaching occurred much faster, within 1-3 hours. Such systems can serve as models for drug delivery agents.

Comparison of four staining methods for detecting eosinophils in nasal polyps.

The study aimed to find a more appropriate method to detect eosinophils in formalin- fixed nasal polyps, since there is no consensus on the standard counting method of eosinophils now. Four 5 µm serial sections were obtained from each 10% neutral formalin-fixed paraffin block and were stained with Chromotrope 2R, Congo red, MBPmAb immunohistochemistry, and conventional hematoxylin and eosin stain respectively. Each section was scanned by the Aperio digital section scanner. The same selected areas were procured for assessment in the serial sections. Chromotrope 2R and MBPmAb immunohistochemistry were specific in detecting eosinophils, which had the lower background staining compared with Congo red and conventional hematoxylin and eosin stain. There were significant differences among the four methods in terms of the eosinophil counting data (p < 0.05), while no significant difference between Chromotrope 2R and Congo red (P = 0.1413). The eosinophil counts in nasal polyps could be more accurately assessed by Chromotrope 2R and Congo red compared with MBPmAb immunohistochemistry and conventional hematoxylin and eosin stain. The popularization of Chromotrope 2R and Congo red may help to unify the eosinophil count in the definition of eosinophilic CRSwNP.

Opposite effects of dextran sulfate 500, the polyene antibiotic MS-8209, and Congo red on accumulation of the protease-resistant isoform of PrP in the spleens of mice inoculated intraperitoneally with the scrapie agent.

The mode and the site of action of the major antiscrapie drugs have been studied by investigating their effects on the abnormal protease-resistant isoform of PrP (PrPres) and on its accumulation in mouse spleen. Day-by-day PrPres accumulation in the spleen and in other peripheral organs was first monitored to describe the early steps of scrapie pathogenesis. Three phases were identified: the detection of scrapie inoculum on the day of scrapie infection, a clearance phase, and then the peripheral accumulation of PrPres. In a second step, the effects of the polyene antibiotic MS-8209, the polyanion dextran sulfate 500 (DS500), and Congo red were assessed on these phases, after the drugs were coincubated with scrapie inoculum. Highly different mechanisms and sites of action were apparent. MS-8209 had a weak effect on the accumulation of PrPres in spleen, suggesting another site of intervention for this drug. DS500 delayed the beginning of the clearance phase but then blocked PrPres synthesis for a long period of time, probably because of its immunological effects on the spleen. Surprisingly, Congo red suppressed the clearance phase of scrapie inoculum and then increased transiently accumulation of PrPres in spleen. We showed in vitro that this effect was related to a direct enhancement of the protease resistance of PrPres by the drug.

Pharmacological manipulation of early PrPres accumulation in the spleen of scrapie-infected mice.

In most experimental models of scrapie and in some naturally infected species, the lymphoreticular system and the spleen in particular play a major role in the pathogenesis of the disease. Previous studies demonstrated scrapie infectivity in peripheral organs from the day of infection up to the terminal stage. The discovery of the abnormal prion protein, PrPres, as a specific molecular hallmark of scrapie should permit enhanced study of scrapie pathogenesis and has some pharmacological applications. In this study, PrPres accumulation was followed day by day in peripheral organs. Four different phases were identified: the circulation of scrapie inoculum, a clearance phase, the peripheral accumulation of PrPres and a plateau phase. This kinetics was then pharmacologically modified (i) by applying the macrophage "suicide" technique to unveil the cellular types involved in scrapie pathogenesis and (ii) with anti-scrapie drugs such as polyene antibiotics, polyanions and Congo red to investigate their mode and site of action.

New chelate-forming polymer microspheres carrying dyes as chelators for iron overload.

Dye-incorporated [poly(EGDMA-HEMA)] microspheres were investigated as a new chelate-forming polymer for iron overload. Poly(EGDMA-HEMA) microspheres, in the size range of 150-200 microm, were produced by a modified suspension polymerization of EGDMA and HEMA. The reactive dye-ligands (i.e. Cibacron Blue F3GA, Alkali Blue 6B and Congo Red) were covalently incorporated to the microspheres. The maximum dye incorporations were 16.5 micromol Cibacron Blue F3GA g(-1), 23.7 micromol Alkali Blue 6B g(-1), and 14.5 micromol Congo Red g(-1). The maximum Fe(III) adsorptions on the dye-incorporated microspheres from aqueous solutions containing different amounts of Fe(III) ions were 51.0, 37.3, and 25.1 mg g(-1) for the Cibacron Blue F3GA, Alkali Blue 6B, and Congo Red carrying microspheres, respectively. The maximum Fe(III) adsorptions were observed at pH 4.0 in all cases. Fe(III) removal from human plasma was also investigated. The maximum adsorption capacities of Fe(III) ions from human plasma for Cibacron Blue F3GA, Alkali Blue 6B, and Congo Red, were of 12.0, 7.5, and 3.8 mg g(-1) polymer, respectively. It was observed that Fe(III) could be repeatedly adsorbed and desorbed without significant loss in adsorption capacity.

Posterior truncal vagotomy and stapling of the anterior stomach wall in 30 patients with duodenal ulcer: acid inhibition, gastric emptying, and endoscopic dye spraying. Prospects for endoscopic vagotomy.

The use of a stapling instrument (TA90) to effect a transection of the anterior gastric wall with posterior truncal vagotomy but preserving innervation to the anterior wall of the antrum for duodenal ulcer is described. The operation resulted in the endoscopic healing of all the ulcers; however, recurrent ulceration was seen in 3 of 25 patients. Basal acid output was reduced from 7.84 (SEM 1.73) to 3.49 (SEM 1.15) mmol/h (t = 2.09, df = 15.7, p = 0.05), while peak acid output was reduced from 38.67 (SEM 3.11) to 19.26 (SEM 2.3) mmol/h (t = 5.01, df = 23.9, p = 0.0000). Solid and liquid gastric emptying studies were performed, and while some delay in solid emptying was seen, only one patient complained of transient gastric stasis. Endoscopic Congo red staining was performed postoperatively and a tongue of innervated mucosa along the lesser curve was seen in each case. The procedure was easily and rapidly performed, and early clinical results are good, with 25 of 30 patients having a good (Visick I) result.

Comportamento morfológico hormonal e secretor gástrico em ulcerosos duodenais submetidos à vagotomia gástrica proximal associada ou näo à secçäo do nervo de Rosati / Morphologic, hormonal and gastric secretores behavior in duodenal ulcer patients subjected to proximal gastric vagotomy associated or not to section of Rosati's nerve

A Vagotomia Gástrica Proximal (VGP) constitui importante altenativa cirúrgica no tratamento da Ulcera Duodenal (UD), sendo a recidiva ulcerosa seu principal efeito indesejável. Na presente pesquisa, objetivou-se avaliar a importância da associaçäo da manobra de Rosati à eventual reduçäo no índice de recidiva ulcerosa após emprego da VGP. Avaliaram-se, para tal, comparativamente 26 pacientes submetidos à VGP por UD, sendo 13 operados pela VGP clásica (VGP-C) e 13 pela VGP ampliada com secçäo do nervo de Rosati (VGP-R). Realizou-se análise estatística (p<0,05) do padräo clínico, morfológico, secretor e hormonal gástrico segundo as seguintes variáveis: 1) Avaliaçäo clínica: 2) Dosagem da secreçäo ácida gástrica em situaçäo basal e após estímulo com pentagastrina; 3) Dosagem do pepsinogênio sérico basal e aos 60 e 90 minutos após mesmo estímulo; 4) Dosagem da gastrinemia basal e após 15, 30, 45 e 60 minutos de estímulo com refeiçäo simulada; 5) Padräo endoscópico pós-operatório; 6) Prova do Vermelho Congo. O seguimento pós-operatório médio foi de 18 meses (12 a 36 meses). Observou-se näo haver diferença estatisticamente significante entre as duas variantes técnicas operatórias em todos os parâmetros estudados. Fez exceçäo a prova do Vermelho Congo, onde os pacientes submetidos a VGP-R apresentaram áreas de viagem significativamente e paradoxalmente maior do que aqueles operados pela técnica clássica, na regiäo da grande curvatura gástrica. Desde que a VGP-R apresentou resultados semelhantes aos da VGP-C, do ponto de vista clínico, morfológico e da secreçäo ácida e hormonal do estômago, no tratamento eletivo da UD crônica, concluiu-se que a associaçäo da manobra de Rosati näo proporciona maior eficiência à VGP, sendo, portanto, procedimento desnecessário

Improved methodology for the Grassi test for intraoperative determination of completeness of vagotomy.

The Grassi test for completeness of vagotomy is a useful adjunct to vagotomy that aids the surgeon in severing all the vagal innervation of the acid-producing portion of the stomach. The Congo Red stick allows the surgeon to perform the Grassi test with a significant reduction in the amount of equipment and personnel required by either pH electrode or liquid Congo Red methods.