Short implants (6mm) vs. vertical bone augmentation and standard-length implants (≥9mm) in atrophic posterior mandibles: a 5-year retrospective study.

This retrospective study compared the 5-year clinical and radiographic outcomes of short implants (6 mm) (short group), and standard-length implants (≥9mm) placed after a vertical augmentation with autologous bone blocks (augmentation group), supporting partial fixed prostheses in the posterior mandible. Forty-five partially edentulous patients were enrolled in the study and evaluated after 5 years: 22 (51 implants) in the augmentation group and 23 (46 implants) in the short group. Eight surgical complications occurred in the augmentation group versus none in the short group (P=0.003). One short implant failed before loading and one standard-length implant failed after 4 years because of peri-implantitis (P=1.0). Eight biological and two prosthetic complications occurred in the augmentation group vs. three biological and three prosthetic complications in the short group (P=0.09 and P=1.0, respectively). A mean marginal bone loss of 1.61±1.12mm in the augmentation group and 0.68±0.68mm in the short group was found (P=0.002). Within the limitations of this study, both techniques resulted in successful clinical results after 5 years, but short implants exhibited less surgical complications and marginal bone loss than standard-length implants placed in augmented bone.

Occurrence of Salmonella, Listeria monocytogenes, and enterotoxigenic Staphylococcus in goat milk from small and medium-sized farms located in Minas Gerais State, Brazil.

Consumption of goat milk has been increasing due to its nutritional characteristics and health benefits. Therefore, assessment of the presence of foodborne pathogens in this product is critical to ensure its safety to consumers. The present study aimed to identify common foodborne pathogens in raw goat milk. Fifty-three samples of raw goat milk from 11 farms were collected and cultured for the presence of Salmonella spp. and Listeria monocytogenes, as well as for enumeration and isolation of coagulase-positive and coagulase-negative Staphylococcus (CPS and CNS, respectively). All samples tested negative for Salmonella spp. and L. monocytogenes. The CPS counts in raw goat milk samples were predominantly less than 2 log cfu/mL (n=39), and CNS counts were predominantly higher than 3 log cfu/mL (n=42). Based on Staphylococcus counts, 51 isolates were selected (CPS=26; CNS=25) and tested by PCR for the presence of classic enterotoxin-encoding genes (sea, seb, sec, sed, and see). Only 3 isolates (CPS=2, CNS=1) were negative for all enterotoxin-encoding genes tested, and the genotype sec and see was the most frequent (n=16), followed by sea, sec, and see (n=13) and sec (n=13); sed was not detected in any isolate. The frequencies of enterotoxin-encoding genes for CPS and CNS were similar, demonstrating the equivalence of both groups in harboring these virulent markers. These results suggest that Salmonella and L. monocytogenes are not frequent contaminants of raw goat milk, but that Staphylococcus spp. that are capable of producing enterotoxins are prevalent; therefore, consumers of raw goat milk and products made from raw milk are at risk.

Spoilage potential of Pseudomonas species isolated from goat milk.

Pseudomonas spp. are usually associated with spoilage microflora of dairy products due to their proteolytic potential. This is of particular concern for protein-based products, such as goat milk cheeses and fermented milks. Therefore, the goal of the present study was to characterize the proteolytic activity of Pseudomonas spp. isolated from goat milk. Goat milk samples (n=61) were obtained directly from bulk tanks on dairy goat farms (n=12), and subjected to a modified International Organization for Standardization (ISO) protocol to determine the number and proteolytic activity of Pseudomonas spp. Isolates (n=82) were obtained, identified by PCR, and subjected to pulsed-field gel electrophoresis with XbaI macro-restriction. Then, the isolates were subjected to PCR to detect the alkaline protease gene (apr), and phenotypic tests were performed to check proteolytic activity at 7°C, 25°C, and 35°C. Mean Pseudomonas spp. counts ranged from 2.9 to 4.8 log cfu/mL, and proteolytic Pseudomonas spp. counts ranged from 1.9 to 4.6 log cfu/mL. All isolates were confirmed to be Pseudomonas spp., and 41 were identified as Pseudomonas fluorescens, which clustered into 5 groups sharing approximately 82% similarity. Thirty-six isolates (46.9%) were positive for the apr gene; and 57 (69.5%) isolates presented proteolytic activity at 7°C, 82 (100%) at 25°C, and 64 (78%) at 35°C. The isolates were distributed ubiquitously in the goat farms, and no relationship among isolates was observed when the goat farms, presence of apr, pulsotypes, and proteolytic activity were taken into account. We demonstrated proteolytic activity of Pseudomonas spp. present in goat milk by phenotypic and genotypic tests and indicated their spoilage potential at distinct temperatures. Based on these findings and the ubiquity of Pseudomonas spp. in goat farm environments, proper monitoring and control of Pseudomonas spp. during production are critical.

Sistemas de efluxo multidrogas em Escherichia coli e uso de inibidores como possíveis adjuvantes na terapia da mastite bovina / Multidrug efflux systems in Escherichia coli and the use of inhibitors as possible adjuvant therapy in bovine mastitis

Vinte e sete isolados de Escherichia coli provenientes de leite de bovinos com mastite clínica foram submetidos a teste de sensibilidade aos principais antimicrobianos usados no tratamento desta doença. Avaliou-se também a efetividade in vitro de dois inibidores de sistemas de efluxo multidrogas, fenilalanina arginyl ß naftilamida (PAβN) e 1-(1-Naphthylmethyl)-piperazine (NMP), utilizando-se a concentração inibitória mínima (CIM) como referência. A CIM e o sistema de efluxo foram detectados com base nas curvas de crescimento, utilizando-se a densidade óptica (D.O550), em diferentes concentrações da droga e na presença e ausência dos inibidores. Apenas quatro isolados apresentaram resistência à ampicilina e ao sulfametoxazol∕trimetoprim, simultaneamente, enquanto todos os 27 foram sensíveis aos demais antimicrobianos. Observaram-se valores para ampicilina variando de 6µg/mL a 250µg/mL e para sulfametoxazol∕trimetoprim de 12µg/mL a 1500µg/mL. Houve redução das CIMs desses antimicrobianos em todos os isolados na presença dos inibidores, exceto para sulfametoxazol∕trimetoprim na presença de NMP. Conclui-se que esses isolados possuem um estreito perfil de resistência e que PAßN apresentou melhor efeito inibitório em relação à ampicilina e ao sulfametoxazol∕trimetoprim, apresentando-se como um candidato a adjuvante no tratamento da mastite...

Twenty-seven isolates of Escherichia coli from cattle with clinical mastitis were subjected to sensitivity tests regarding main antimicrobials used in the treatment of this disease. We also evaluated in vitro effectiveness of two inhibitors of multidrug efflux systems, fenilalanina arginyl ß naftilamida (PAβN) and 1-(1-Naphthylmethyl)-piperazine (NMP), using the minimum inhibitory concentration (MIC) as a reference. MIC and multidrug efflux systems were detected in the growth curves, using optical density (D.O550) at different drug concentrations and the presence and absence of inhibitors. Only four isolates of E. coli were resistant to ampicillin and trimethoprim/sulfamethoxazole, simultaneously. However, all isolates were sensitive to the other antimicrobials. Were observed values ranging from 6mg to 250mg ampicillin/mL, and 12mg to 1500mg/mL trimethoprim/sulfamethoxazole. There was a reduction of the MIC of antimicrobials for all isolates in the presence of inhibitors, except for trimethoprim/sulfamethoxazole in the presence of NMP. In conclusion, these isolates have a narrow resistance profile and PAßN showed better inhibitory effect against ampicillin and trimethoprim/sulfamethoxazole, and is a candidate for the adjuvant treatment of mastitis...

Perfil de sensibilidade de células sésseis e planctônicas de Escherichia coli a antimicrobianos usados no tratamento da mastite bovina / Sensitivity profile of planktonic and sessile cells of Escherichia coli to antimicrobial used in the treatment of bovine mastitis

Escherichia coli é um micro-organismo altamente adaptativo e sua habilidade em formar biofilmes pode ser fundamental na resistência a tratamentos com antimicrobianos. A avaliação da concentração mínima inibitória (CMI) vem sendo utilizada para verificar a sensibilidade dos micro-organismos aos antimicrobianos. Entretanto, quando se avaliam células sésseis, a concentração do antimicrobiano requerido para erradicação do biofilme é maior do que a determinada pela CMI. Objetivou-se comparar as CMI com as concentrações mínimas de erradicação de biofilmes (CMEB) de antimicrobianos usados no tratamento da mastite em 27 isolados de E. coli produtores de biofilmes provenientes de mastite. Os isolados foram submetidos a testes de sensibilidade a antimicrobianos usados no tratamento da mastite, tanto para células planctônicas, por meio da CMI, quanto para células sésseis, pela avaliação da CMEB. Os resultados revelaram uma alta sensibilidade: apenas quatro (14,8%) isolados obtiveram valores da CMI elevados, variando de 4 a 10µg/mL, sendo classificados como resistentes. Para os demais isolados (85,2%), os valores foram menores, variando de 0,125 a 2µg/mL, classificados como sensíveis. A avaliação de CMEB indicou que a concentração dos antimicrobianos necessária para eliminar as células sésseis variou de 100µg/mL a 500µg/mL. Os valores de CMEB foram significativamente maiores nos isolados grandes e moderados produtores de biofilmes em relação aos isolados fracos produtores de biofilmes (p<0,001). Não houve correlação entre os valores de CMEB e CMI (p>0,05). A escolha da terapêutica antimicrobiana correta para o tratamento de infecções intramamárias em bovinos relacionadas com a produção de biofilmes parece exigir a aplicação de testes mais específicos. Testes de susceptibilidade antimicrobiana baseados apenas em valores de CMI mostraram-se ineficazes em determinar com precisão a susceptibilidade das células bacterianas sésseis.

Escherichia coli is a highly adaptive microorganism. Its ability to form biofilms may be critical for resistance to antimicrobial treatments. Evaluation of minimum inhibitory concentration (MIC) has been used to check the sensitivity of microorganisms to antibiotics, however, when evaluating sessile cells, the required antibiotic concentration to eradicate biofilm is greater than determined by MIC. This study aimed to compare MIC with minimum biofilm eradication concentration (MBEC) of antimicrobials used in mastitis treatment in 27 E. coli biofilm producers isolates from mastitis. Isolates were tested for sensitivity to antimicrobials used in mastitis treatment, for both planktonic cells (by CMI) and sessile cells (by MBEC). The results revealed high sensitivity: only four (14.8%) isolates showed high MIC values, ranging from 4 to 10g/mL and they were classified as resistant. All other isolates (85.2%) showed lower values, ranging from 0.125 to 2mg/mL, and they were classified as sensitive. Evaluation of MBEC indicated that concentration of antimicrobial needed to remove sessile cells ranged from 100mg/mL to 500mg/mL. MBEC values were significantly higher in large and moderate biofilm producers isolates regarding weak biofilm producers isolates (p<0.001). There was no correlation between MBEC and CMI values (p>0.05). The correct choice of antimicrobial therapy for treatment of mammary infections in cattle related to biofilm production seems to require application of more specific tests. Antimicrobial susceptibility testing based only on MIC values proved ineffectiveness to accurately determination the susceptibility of sessile bacterial cells.

Occurrence of linear and cyclic volatile methyl siloxanes in indoor air samples (UK and Italy) and their isotopic characterization.

The occurrence of linear- and cyclic-volatile methyl siloxanes (lVMSs and cVMSs, respectively) in various indoor environments, occupational and domestic, in Italy and in the United Kingdom was studied. The results show that the cVMSs are the most abundant, detected in average concentrations that in some cases were as high as 170µgm(-3). Our study highlights the differences that can be observed between various indoor environments (e.g. domestic like bathrooms, bedrooms, or occupational) and between two countries. In most cases, the concentrations found in the UK are higher than in the respective indoor environments in Italy. The assessment of exposure to these two countries for adults and children revealed significant differences both not only in the levels of exposure, but also in the patterns. In Italy, the biggest part of the exposure to VMSs takes place domestically, whereas in the UK, it is observed for occupational environments. Additionally, the compound specific isotopic analysis was employed as a source identification technique. The results are promising mainly for D5 that occurs in higher concentrations, but not for the less abundant lVMSs and cVMSs.

Antimicrobial profile screening of two oils of Copaifera genus / Avaliação do perfil antimicrobiano de dois óleos do gênero Copaifera

O objetivo deste estudo foi identificar a atividade inibitória de óleos de copaíba sobre o crescimento dos micro-organismos: Shigella flexneri, Klebsiella pneumoniae, Escherichia coli, Proteus mirabilis, Pseudomonas aeruginosa, Citrobacter freundi, Staphylococcus aureus, Actinobacillus pleuropneumoniae e Haemophilus parasuis. Foi realizado um teste de difusão em ágar com duas soluções a 10 por cento de óleos de copaíba obtidos de duas diferentes espécies de copaíba (Copaifera officinalis e C. langsdorffii) e um controle negativo com tween 80 e água. Os isolados clínicos de H. parasuis e A. pleuropneumoniae foram incubados em microaerofilia. Os resultados mostraram três espécies Gram-negativas inibidas por ambas as soluções de óleo de copaíba: E. coli, P. aeruginosa e S. flexneri. Na inibição de P. aeruginosa o óleo de C. officinalis foi superior ao de C. langsdorffii (P<0,05). Todas as cepas de S. aureus tiveram seu crescimento inibido pelas soluções no ensaio, sem diferença estatística entre os halos. Estes resultados sugerem que o óleo de copaíba pode ser uma fonte potencial de compostos inibitórios para ser utilizada como antimicrobianos no tratamento de infecções humanas e animais e conservação de alimentos.

Levantamento sorológico de Mycobacterium avium subesp. paratuberculosis em bovinos leiteiros no estado do Espírito Santo / Serological survey of Mycobacterium avium subsp. paratuberculosis in dairy cattle in Espírito Santo state

The occurrence of antibodies to Mycobacterium avium subsp. paratuberculosis (MAP) was verified in dairy cattle from Espírito Santo state. A total of 1,450 serum samples were analyzed for antibodies anti-MAP, using ELISA. Dairy cattle, males and females, from four regions of Espírito Santo state were used. One hundred sixty-five (11.4 percent) samples were positive for anti-MAP, 33 (2.3 percent) were considered suspicious, and 1,252 (86.3 percent) were negative. In all regions, seropositive animals were found, indicating that the agent is spread by the State, posing a threat to the local dairy farming and neighboring states, as well as public health, since MAP can be involved with Crohn's disease in humans. This result presents the first serologic anti-MAP survey in dairy cattle of Espírito Santo State.

Efeitos clínicos e microbiológicos do óleo de copaíba (copaifera officinalis) sobre bactérias formadoras de placa dental em cães / Clinical and microbiological effects of copaiba oil (copaifera officinalis) on dental plaque forming bacteria in dogs

O potencial de uso do óleo de copaíba (Copaifera officinalis) na prevenção da doença periodontal, eliminando seu agente etiológico, foi avaliado em 18 cães sem raça definida, distribuídos homogeneamente em três grupos: teste, (contendo óleo de copaíba) controle positivo e controle negativo. Os tratamentos ocorreram três vezes ao dia, durante oito dias. Ao nono dia, os animais receberam aplicação tópica de fucsina básica 0,5 por cento para evidenciação do biofilme. Mudanças na halitose e gengivite foram avaliadas diariamente por inspeção visual. Adicionalmente, foram realizados testes laboratoriais de inibição de aderência de Streptococcus mutans e ensaio antimicrobiano de difusão em ágar, sobre bactérias formadoras de placa dental. Os resultados da placa evidenciada apontaram áreas de cobertura microbiana nos dentes de 53,4±8,8 por cento, 28,5±5,4 por cento, e 22,3±5,3 por cento para os grupos negativo, positivo e teste, respectivamente, indicando diferença entre o controle negativo e os demais grupos (P<0,05). Quanto à melhora nos aspectos clínicos, halitose e gengivite, o grupo teste respondeu melhor quando comparado ao grupo controle negativo (P<0,05). A análise dos ensaios de difusão e inibição de aderência mostrou superioridade do grupo da copaíba (teste) em relação aos outros grupos (P<0,05). Os resultados sugerem o uso do óleo de copaíba na prevenção da doença periodontal e como um possível substituto da clorexidina na terapia antimicrobiana oral.

The copaiba oil (Copaifera officinalis) potential was evaluated in preventing periodontal disease and reducing its etiology. For that 18 mongrel dogs were homogeneously distributed in three groups: test (copaiba oil), positive control (chlorexidine) and negative control. The treatments were carried out three times a day, during eight days. On the 9th day, the animals were tested with a 0.5 percent basic solution of fuchsin for the detection of biofilm. Changes in halitosis and gingivitis were daily observed. In addition, the following laboratory tests were done: inhibition of the adherence of Streptococcus mutans, and plaque forming bacteria antimicrobial assays by the agar diffusion method. The results of the fuchsin test showed that dental plaque reached areas of 53.4±8.8 percent, 28.5±5.4 percent, and 22.3±5.3 percent in the negative control, positive control, and test groups, respectively, showing differences between dogs from the negative control group and dogs from the other two groups (P<0.05). Furthermore, halitosis and gingivitis decreased in the copaiba group animals when compared with the negative group (P<0.05). The results of the attachment inhibition and agar diffusion tests showed that copaiba induced better effects against the microorganisms as compared to the results of the other groups (P<0.05). These findings suggest that copaiba oil may effectively replace chlorexidine for oral antimicrobial therapy and prevention of periodontal disease.

Trans-tracheostomic endoscopy of the larynx in the evaluation of dysphagia.

Laryngeal endoscopy plays a determinant role in clinical evaluation of dysphagia. The examination is performed by means of a trans-nasal approach with a flexible fiberoptic endoscope, able to visualize the pre- and post-deglutitory steps of the pharyngeal phase of swallowing. In patients with tracheostomy, it is possible to visualize the glottic or neoglottic function during the intra-deglutitory phase, performing the examination through a trans-tracheostomic route. The procedure and indications of this endoscopic technique are described.

Surgical rehabilitation of dysphagia after partial laryngectomy.

Surgical rehabilitation of dysphagia in patients who have undergone partial laryngectomy is aimed at improving the sphincteric action of the larynx, the anatomy and physiology of which are impaired. The interventions indicated for this purpose can be performed either with an external or endoscopic approach. The Authors present early results of their experience employing injection laryngoplasty with autologous fat, bovine collagen (Zyplast, McGhan Medical Corporation, Fremont, CA, USA) and PDMS (Vox Implants, Uroplasty Inc, Minnetonka, MN, USA), performed by means of fiberendoscopy, under local anaesthesia, and microlaryngoscopy, under general anaesthesia.

[Scabies in a dalysis unit. Mystery and prejudice]. / La scabbia in un centro dialisi. Mistero e pregiudizio.

BACKGROUND: Pruritus is one of the most common symptoms of uremia. The etiology of uremic pruritus is steel incompletely known. Scabies may be a cause of itching in these patients. METHODS: Retrospective analysis of beginning and developing of a scabies outbreak in a Dialysis Unit with 160 patients. RESULTS: Sixteen cases of scabies were observed in the Dialysis Unit from April 1998 and January 1999: in 6 of them the scarification was positive. In 10 the scarification was negative, but itching disappeared after treatment with benzyl benzoate 20%. Many courses were necessary (max 6). Prophylaxis (treatment with benzyl benzoate 20% for 3 days and lingery cleaning) was applied to approximately 400 people. No cases were observed among health care workers of the Dialysis Unit. CONCLUSIONS: In a Dialysis Unit the diagnosis of scabies is difficult because the patients often have generalized itching; moreover some of them are affected by neuropathy that may make the infestation of scabies more difficult to identify. The most important factor to limitate the outbreak seems to be the prophylaxis of people who take care of patients (health-care workers, family members and car-drivers). It seems also necessary to repeat the treatments many times. The most exposed patients seemed to be those with diminished independence, diabetes and malnutrition.

[Helicobacter heilmannii: anatomo-clinical study of 14 new cases]. / Helicobacter heilmannii: studio anatomo-clinico di 14 nuovi casi.

INTRODUCTION: Helicobacter pylori is one of the most common causes of human gastritis. Recently, a new agent has been isolated, which also causes a gastritis. It has been initially named Gastrospirillum hominis and renamed Helicobacter heilmannii (Hh). Hh is extremely rare. In spite of the rarity it is important to recognize and diagnose it, as it requires a proper therapy, different from Hp therapy. Clinical presentation and serological results of Hh are superimposable to those of HP. Therefore differential diagnosis resides on histological grounds. PURPOSE of the present paper is to report 14 new cases of Hh gastritis, which constitutes the first italian series. RESULTS: Cases constituted 0.01% of all gastric biopsies seen in the period 1994-1998. Nine patients were male and five were female; age ranged from 32 to 76 years (50 years on average). All patients presented a mild to moderate gastritis. Hh is a spiral bacterium, being about 10 micra in length, localized in single or small groups in the glandular mucus. Two cases were associated with Hp. One case was associated with gastric adenocarcinoma. Two cases were diagnosed during the follow-up of duodenal ulcer. In CONCLUSION, the incidence of Hh gastritis in the present series seems consistent with that from other European countries. In all cases the presence of Hh was associated with features of gastritis. This confirms the pathogenetic role of Hh.

The changing face of AIDS-related opportunism: cryptococcosis in the highly active antiretroviral therapy (HAART) era. Case reports and literature review.

Only nine cases of AIDS-related cryptococcosis have been reported until now in patients receiving highly active antiretroviral therapy (HAART), all of them with abnormal clinical features. Two HIV-infected patients who experienced an atypical relapse of cryptococcosis shortly after the start of HAART and despite maintenance antifungal treatment, are described. Six different relapses of cryptococcal meningitis were observed in a 28-month period in a patient who obtained a poor immune recovery after HAART (as shown by a CD4+ lymphocyte count ranging from 78 to 149 cells/microL, opposed to a baseline level of 98 cells/microL). On the other hand, a patient with favorable immunological response to HAART (as expressed by a CD4+ count growing from 7 to 186 cells/microL), experienced isolated multiple indolent cryptococcal abscesses involving head, neck, the anterior thoracic wall, and regional lymph nodes, with repeatedly negative cultures, and diagnosis obtained by both histopathologic study and positive serum antigen assay. Both our case reports are representative of novel correlations between opportunistic pathogens and immune reactivity, descending from the introduction of HAART. The first episode describes an exceedingly elevated number of disease relapses despite HAART and antifungal maintenance treatment, which may descend from an incomplete immune response to antiretroviral therapy, possibly responsible for failure in obtaining eradication of yeasts, but also for lack of disease dissemination (usually leading to a lethal multivisceral involvement in the pre-HAART era). The abnormal disease course and localization of second reported patient well depicts an "immune reconstitution syndrome" probably representing a flare-up of a latent fungal infection, caused by a rapidly effective HAART. In patients treated with HAART, AIDS-related cryptococcosis cannot therefore be ruled out by the absence of neurological involvement, and by persistingly negative cultures.

The EnVision++ system: a new immunohistochemical method for diagnostics and research. Critical comparison with the APAAP, ChemMate, CSA, LABC, and SABC techniques.

AIM: To assess a newly developed immunohistochemical detection system, the EnVision++. METHODS: A large series of differently processed normal and pathological samples and 53 relevant monoclonal antibodies were chosen. A chessboard titration assay was used to compare the results provided by the EnVision++ system with those of the APAAP, CSA, LSAB, SABC, and ChemMate methods, when applied either manually or in a TechMate 500 immunostainer. RESULTS: With the vast majority of the antibodies, EnVision++ allowed two- to fivefold higher dilutions than the APAAP, LSAB, SABC, and ChemMate techniques, the staining intensity and percentage of expected positive cells being the same. With some critical antibodies (such as the anti-CD5), it turned out to be superior in that it achieved consistently reproducible results with differently fixed or overfixed samples. Only the CSA method, which includes tyramide based enhancement, allowed the same dilutions as the EnVision++ system, and in one instance (with the anti-cyclin D1 antibody) represented the gold standard. CONCLUSIONS: The EnVision++ is an easy to use system, which avoids the possibility of disturbing endogenous biotin and lowers the cost per test by increasing the dilutions of the primary antibodies. Being a two step procedure, it reduces both the assay time and the workload.

[Effects of alpha and beta blocker properties of parahydroxybenzonitrile on the cardiovascular system of rats]. / Etudes des effets alpha et beta bloquants du parahydroxybenzonitrile sur le système cardiovasculaire du Rat.

The study of parahydroxybenzonitrile on the cardiovascular system of the Rat shows alpha blocker properties.

Concurrence of localized Castleman's disease and peripheral small B-Lymphocytic lymphoma within the same lymph node.

Castleman's disease, also known as benign giant lymph node hyperplasia, is a lymphoproliferative disorder which can occur either in a localized or multicentric form. The latter is characterized by the development of malignant lymphoma, Kaposi's sarcoma or carcinoma in 32% of cases. By contrast, localized Castleman's disease has exceptionally been reported in association with non-Hodgkin's lymphoma and - to the best of our knowledge - never at the same anatomic site. Here we describe the occurrence in the same lymph node of localized Castleman's disease (with monotypic plasma cell component) and an apparently unrelated peripheral small B-lymphocytic lymphoma.

Piperazine derivatives of butyric acid as differentiating agents in human leukemic cells.

Butyric acid is a potent antineoplastic agent with a well-documented differentiation activity on a wide variety of tumor cells. However, its clinical development is strongly limited by its very short metabolic half-life. In this study we report on the in vitro effects of new original piperazine derivatives of butyric acid on the induction of differentiation and the growth inhibition of human erythroleukemia K562 cells and myeloid leukemia HL60 cells. 1-(2-hydroxyethyl) 4-(1-oxobutyl)-piperazine (HEPB) and [1-(2-hydroxyethyl) 4-(1-oxobutyl)-piperazine] butyrate (HEPDB) were efficient in acting on the differentiation and proliferation of both cell lines, whereas 1-phenyl 4-(1-oxobutyl)-piperazine (PPB) and 1-(3,4-methylene dioxybenzyl) 4-(1-oxobutyl)-piperazine (POB) acted only on proliferation rates. Such derivatives did not induce significant toxicity in mice. These preliminary results should enable, by the development of new series of piperazine derivatives, a better understanding of the mechanisms of action of butyric acid and its analogues on the coupling of growth and differentiation of neoplastic cells.

Antigen retrieval techniques in immunohistochemistry: comparison of different methods.

Routine sections of normal and pathological samples fixed in 10 per cent buffered formalin or B5, including EDTA-decalcified bone-marrow biopsies, were tested with 61 antibodies following heating in three different fluids: 0.01 M citrate buffer (pH 6.0), 0.1 M Tris-HCl (pH 8.0), and 1 mM EDTA-NaOH solution (pH 8.0). The sections underwent either three cycles of microwave treatment (5 min each) or pressure cooking for 1-2 min. The alkaline phosphatase/anti-alkaline phosphatase (APAAP) technique was used as the standard detection method; with 16 antibodies a slightly modified streptavidin-biotin complex (SABC)-immunoperoxidase technique was applied in parallel. The results obtained were compared with those observed without any antigen retrieval (AR), or following section digestion with 0.05 per cent protease XIV at 37 degrees C for 5 min. Chess-board titration tests showed that all antibodies but one profited by AR. Protease XIV digestion represented the gold standard for five antibodies, while 55 produced optimal results following the application of heat-based AR. By comparison with the other fluids, EDTA appeared to be superior in terms of both staining intensity and the number of marked cells. These results were independent of tissue processing, immunohistochemical approach, and heating device. Pressure cooking was found to be more convenient on practical grounds, as it allowed the simultaneous handling of a large number of slides and a time saving of 1 min 30 s, representing the proper time for the treatment.

Peripheral T-cell lymphomas. Clinico-pathologic study of 168 cases diagnosed according to the R.E.A.L. Classification.

BACKGROUND: One hundred sixty-eight peripheral T-cell lymphomas (PTCLs) were reviewed according to the Revised European-American Lymphoma (R.E.A.L.) Classification. PATIENTS AND METHODS: The cases, originally diagnosed on the basis of the Updated Kiel Classification (UKC), were all provided with histological preparations, immunophenotype, clinical information, and follow-up data. The slides were reclassified by five observers, who integrated the R.E.A.L. criteria with cell size measurements. The prognostic value of clinical and pathologic findings was assessed by univariate and multivariate analysis. RESULTS: The R.E.A.L. Classification was reproducibly applied by all of the observers. Clinically, anaplastic large cell lymphomas (ALCLs) differed from the remaining PTCLs by mean age (29.5 vs. 52.9 years), bulky disease (52.3% vs. 11.3%; P = 0.000), mediastinal mass (52.7% vs. 32%; P = 0.004), and disease-free survival (68.0% vs. 38.2%; P = 0.0001). Although each histological type displayed specific clinical aspects, PTCLs other than ALCL were basically characterised by a poor clinical outcome which was not influenced by the UKC malignancy grade. At multivariate analysis, the risk of a lower complete remission rate was related to bulky disease (P = 0.001), histologic group (non-ALCL) (P = 0.01), and advanced stage (III-IV) (P = 0.0002). CONCLUSIONS: The present study supports the classification of T-cell lymphomas proposed by the R.E.A.L. scheme.