BKV reactivation in renal transplant recipients: diagnostic and therapeutic strategy--case reports.

The Polyomaviridae family includes several viruses that are ubiquitous with specific host spectra. The human polyoma viruses BK and JC were discovered in 1971. Following primary infection, transmitted by the respiratory and probably the oral route, BK remains latent in uroepithelial cells, in B lymphocytes, or in other tissues (spleen, brain). Reactivation with asymptomatic viruria may occur in both immunocompetent subjects and immunocompromised patients. In renal transplant recipients, BKV replication may cause tubulointerstitial nephropathy (BKVAN) with increasing prevalence rates--1% in 1995, 8% in 2007--leading to the loss of the transplanted organ in 30% to 80% of cases. With the availability of diagnostic programs (decoy cells in urine, amplification of viral DNA by polymerase chain reaction (PCR) on serum and urine, real time (RT)-PCR test for mRNA VP1 urine (mRNA-VP1), and renal biopsy accompanied by reduction in immunosuppression, administration of leflunomide, cidofovir (after hydration), and N-acetylcysteine, as well as immunoglobulin by intravenous injection (IVIg), the incidence of renal loss caused by BKVAN infection has been reduced by 10% to 80%. In this study, we have described 12 patients: 6 treated with tacrolimus (FK), mycophenolate mofetil (MMF), and steroids, and 6 treated with cyclosporine or with mTOR inhibitors. Two patients from the first group showed BKVAN about 3 months posttransplantation. Early diagnosis and therapeutic intervention (cidofovir + IVIg) led to reduction in the viral load, with improvement and stabilization in renal function. Considering the high positive predictive value (98%) of mRNA VP1, it should be possible to avoid renal biopsy. The level of immunosuppression--rather than the immunosuppressive drug itself (FK and MMF)--seemed to be associated with BKV reactivation.

Proteolysis in miniature cheddar-type cheeses manufactured using extracts from the crustacean Munida as coagulant.

Miniature (20 g) Cheddar-type cheeses were manufactured using enzymes extracted from the crustacean Munida or chymosin as coagulant. Cheeses were ripened at 8 degrees C and samples were collected for analysis after 2, 6 and 12 weeks. Proteolysis was assessed by urea-polyacrylamide gel electrophoresis, which showed that cheeses manufactured with the Munida extracts had a higher extent of degradation of beta-casein than cheeses made using chymosin as coagulant. Patterns of proteolysis were also obtained by reverse-phase high-performance liquid chromatography (RP-HPLC) and matrix assisted laser desorption ionisation-time of flight (MALDI-ToF) mass spectrometry. In general, the products of proteolysis were more complex in cheese made using the Munida extracts than in cheese made by chymosin as coagulant. Statistical analysis of results clearly discriminated the cheeses on the basis of coagulant used. Molecular mass of peptides found in cheese made using Munida extracts were similar to those of peptides commonly detected in cheeses made using chymosin as coagulant.

[A vaccination campaign for gypsy children in Rome]. / Una campagna vaccinale in favore dei bambini Rom e Sinti a Roma.

Several studies conducted in Rome have shown low levels of vaccination coverage in gypsy communities. Thus a vaccination campaign targeting to 2400 gypsy children aged 0-13 years, present in 32 settlements in Rome, was conducted in 2002. The campaign was related to vaccinations required and recommended in Italy: diphtheria, tetanus, hepatitis B, pertussis (whooping cough), haemophilus influenzae and measles. In the majority of cases it was decided to carry out the vaccinations directly in the gypsy settlements, in accordance with the methods of pulse immunisation. In the case of small settlements a strategy for reorienting the population to vaccination centres was adopted. Around 2000 children were vaccinated, equivalent to 80% of the paediatric population present during the period. The number of children who have never been vaccinated has decreased from 40% prior to the campaign to 9% after the third week. Vaccination coverage in medium- and small-sized settlements (<200 inhabitants) after the campaign shows values of over 70%; in the large settlements, more modest increases have been recorded and coverage has rarely exceeded 50%. This experience has highlighted the importance of networking between public healthcare institutions and non-profit organisations. The mobilisation of a wide range of competences has thus enabled the attainment of a high level of effectiveness.

Extracting and purifying R-phycoerythrin from Mediterranean red algae Corallina elongata Ellis & Solander.

R-Phycoerythrin (R-PE) is a protein acting as a photosynthetic accessory pigment in red algae (Rodophyta). This protein has gained importance in many biotechnological applications in food science, immunodiagnostic, therapy, cosmetics, protein and cell labelling, and analytical processes. In this paper we report on a new, one step procedure for the extraction and purification of R-PE from a new source: the Mediterranean red algae Corallina elongata Ellis & Solander. This red algae contains mainly R-PE and is suitable for the production in culture. No other contaminating phycobiliproteins could be detected in the extracts. The method we propose for the purification is based on the use of hydroxyapatite, a chromatographic resin that can be produced in the laboratory at very low cost and can be used batch-wise with large amounts of extracts, alternative to chromatography, and therefore can be scaled up. Both the yield and the purity of R-PE are very good.

[Hemodiafiltration with endogenous reinfusion in chronic inflammation: a possible therapeutic alternative?]. / L'impiego dell' Emodiafiltrazione con reinfusione endogena (HFR) nell'infiammazione cronica idiopatica: una possibile alternativa terapeutica?

The patients under maintenance haemodialysis (HD) continue to have an unacceptably excess of mortality compared to general population, that may be explained by high prevalence of inflammation that significantly influences the survival of these patients. Indeed, chronic inflammation is very common in HD and it may cause malnutrition and progression of atherosclerotic disease by several pathogenetic mechanisms triggered by pro-inflammatory cytokines. Currently no pharmacological intervention is specifically targeted the idiopathic chronic inflammation. Hemodiafiltration with endogenous reinfusion (HFR) is a dialysis technique, highly biocompatible, that combines three depurative mechanisms: diffusion, convection and absorption. The ultrafiltrate is obtained from convective section of dialyzer (convection). It is regenerated by passing through the adsorbent macro-porous synthetic resin cartridge (absorption) and then it is reinfused into the second section of the filter (diffusion). This resin cartridge is able to absorb cytokines and other uremic toxins, whereas allows to pass nutrients and antioxidants, as amino acids and vitamins, with a consequent decrement of inflammation and oxidative stress. These characteristics suggest the use of HFR in HD patients affected by overt and idiopathic chronic inflammation. In these patients, we observed that the switching from Bic-HD to HFR allowed an improvement of inflammatory as testified by a significant decrement of serum levels of CRP IL-6, IL-1 and TNF- and a significant increase of albumin and pre-albumin. Whether these favorable effects may modify the outcomes of these high-risk patients, needs to be confirmed by studies ad-hoc.

Cardiovascular risk factors in renal transplant patients after switch from standard tacrolimus to prolonged-release tacrolimus.

Cardiovascular (CV) diseases are the leading cause of death after renal transplantation. Renal transplant patients present CV risk factors that correlate with renal function and the use of immunosuppressive drugs. Noncompliance with immunosuppressive therapy after organ transplantation increases the incidence of rejection, graft loss, and patient death. A simple posology regimen is the best way to promote compliance with prescribed therapy. To meet this need, a new formulation of tacrolimus that is suitable for once-daily administration, is now available on the market: prolonged-release tacrolimus (Fkpr). We analyzed changes in CV risk factors observed in renal transplant patients after transition from standard tacrolimus (Fk) to Fkpr and the rate of patients with the investigated parameters within the normal ranges before and after conversion. The study enrolled 40 Caucasian renal transplant patients (26 men and 14 women) who were being followed at our posttransplantation day hospital clinics. After a varying time interval after transplantation, patients on treatment with tacrolimus, mycophenolate + mofetil (MMF), and steroid entered a 12-month observation period. Thereafter, they were switched to Fkpr, also in association with MMF and steroid, and were observed for a further 12-month period. The following parameters were tested in all patients: creatinine, creatinine clearance, insulin resistance, total cholesterol, HDL cholesterol, LDL cholesterol, triglycerides, uric acid, homocysteine, and urine magnesium. The switch from Fk to Fkpr showed an improvement of the parameters investigated. Moreover, the proportion of patients with normal laboratory values after the transition from Fk to Fkpr was noted either to increase or to remain stable at the improved levels observed during therapy with Fk. Immunosuppressive treatment with Fkpr represents an even better option than Fk for renal transplant patients, because by reducing CV risk factors it favorably affects the long-term outcomes for graft and patient.

Immunosuppressive agents and bone disease in renal transplant patients with hypercalcemia.

Renal transplantation is the definitive treatment for many metabolic abnormalities of uremic patients, although it is only partially effective for renal osteodystrophy, which may interact with posttransplant renal osteopathy. Osteopenic-osteoporotic syndrome represents, together with fractures secondary to osteoporosis and osteonecrosis, the bone complication most related to renal transplantation. Several factors contribute to the pathogenesis of posttransplantation osteoporosis, particularly immunosuppressive treatment. In this study, we evaluated the prevalence of factors related to posttransplant renal osteopathy and the clinical impact of immunosuppressive protocols. We studied 24 renal transplant recipients with hypercalcemia. Glomerular filtration rate was >50 mL/min. Mean age, time on dialysis, and time from transplantation were 49.6, 5.4, and 6.9 years, respectively. We evaluated serum and urine calcium and phosphorus, calcitonin, parathormone, bone-specific alkaline phosphatase, osteocalcin, urine deoxypyridinoline, telopeptide of type 1 procollagen, 1,25-(OH)(2) and 25-OH vitamin D, parathyroid ultrasound, and computerized bone mineralometry. The combination of sirolimus and steroids resulted in the most disadvantageous outcomes regarding alkaline phosphatase and mineralometry. Calcineurin inhibitors did not significantly influence bone metabolism markers; mycophenolate mofetil evidenced no effect on bone. According to the literature, steroids account for the abnormalities found in our patients and in severe osteopenia. Several factors may contribute to the development of osteoporosis and fractures in transplantation patients, although they are overcome by the prominent effect of steroids. In patients at high risk of osteoporosis, steroid-free therapy should be considered. Everolimus is indicated for diseases with bone loss. Combined therapy with everolimus and mycophenolic acid without cyclosporine and steroids, seemed to be particularly indicated. Prophylactic treatments should be commenced early. No single marker was useful to diagnose posttransplant renal osteopathy. The definitive diagnosis should be made by bone biopsy during transplantation, and noninvasive procedures, such as densitometry and evaluation of biologic markers, may be useful during follow-up.

Immunosuppressive agents and metabolic factors of cardiovascular risk in renal transplant recipients.

Cardiovascular disease (CVD) accounts for 35% to 50% of deaths among renal transplant recipients. Beside the atherogenic risk factors related to hemodialysis, renal function, and use of immunosuppressive agents, other relevant risk factors for CVD include acute rejection episodes, microalbuminuria (muAlb), diabetes, arterial hypertension, lipid disorders, inflammatory triggers, hyperhomocysteinemia, anemia, erythrocytosis, obesity, and hyperuricemia. We studied the prevalence of risk factors and the impact of various drugs on CVD among 103 renal transplant recipients with measured glomerular filtration rates showing values >45 mL/min. We measured uric acid, triglycerides (TG), low-density lipoprotein (LDL)/high-density lipoprotein (HDL) LDL/HDL ratio, homocysteine (HOMO), insulin resistance, muAlb, C-reactive protein (CRP), and fibrinogen. Subsequently, patients were divided into 8 groups based on the immunosuppressive protocol to evaluate its impact on CVD risk factors. Insulin resistance and hyperhomocysteinemia were present in >2/3 of patients. Considering the impact of protocols, the combination of cyclosporine (CsA) + everolimus (EVL) resulted in the most favorable profile in terms of reduction of hyperuricemia, hyperlipidemia, and hyperhomocysteinemia. Insulin resistance tended to be more frequent among patients treated with protocols including calcineurin inhibitors (CNI) and steroids. The prevalence of hyperhomocyteinemia was similar among patients on CsA and on tacrolimus (Tac). Sirolimus (SRL) was associated with higher levels of HOMO. The combination of CNI and proliferative signal inhibitors (PSI) seemed to be the most promising one to reduce the impact of CVD risk factors. The reduction in CVD morbidity can improve expectancy and quality of life, as well as graft function and survival among renal transplant patients.

One-step separation from lactose: recovery and purification of major cheese-whey proteins by hydroxyapatite--a flexible procedure suitable for small- and medium-scale preparations.

The recovery of cheese-whey proteins and lactose represents an important task both in environmental and in food sciences. Optimization of whey processing requires the quantitative separation of whey proteins from lactose, lower costs, harmless environmental impact, flexibility in protein recovery, and adaptability of the process to type and amount of available whey. Here we present a method based on the use of self-made, low-price, and nontoxic hydroxyapatite for one-step separation of lactose (non adsorbed) from bovine whey proteins (adsorbed). Recovery of proteins can be performed with high flexibility. Total protein fraction can be eluted with 0.4 M phosphate at pH 7.0. In alternative, proteins can be recovered in pairs with 0.4 M phosphate but at different pH's. About 56% of the proteins, primarily alpha-lactalbulmin and IgG, were eluted at pH 5.0. The other major proteins, beta-lactoglobulin and BSA, were eluted at pH 6.0. Fractions eluted with the two first eluants at pH 5.0 and pH 6.0 were applied to a Superdex 75 column for final purification by gel filtration. This method provides flexibility in whey protein recovery and quantitative separation of proteins from lactose before ultrafiltration and nanofiltration.

Comparação entre óleo de borragem e óleo de girassol quanto à eficácia no tratamento de mulheres com mastalgia cíclica / Comparison of borage oil and sunflower oil according to the efficacy for the treatment of women with cyclical mastalgia

A mastalgia cíclica constitui freqüente e exagerado processo fisiológico, recorrente na fase lútea do ciclo menstrual. Quando intensa, passa a comprometer a qualidade de vida da mulher, por interferir em suas atividades cotidianas. Com o objetivo de determinar, comparativamente, a efetividade e os possíveis efeitos dos óleos de borragem e de girassol, no tratamento da mastalgia cíclica moderada à intensa, foi realizado um estudo aleatório, duplo cego, tipo ensaio clínico, no período de fevereiro de 2001 a dezembro de 2002, envolvendo 111 mulheres. Mensalmente, a intensidade de dor mamária foi avaliada por escala visual analógica (EVA). As participantes foram subdivididas em dois grupos, segundo medicamento administrado, a saber, 56 em uso de cápsulas de óleo de borragem e 55 de óleo de girassol, ambos na dose de 1 cápsula ao dia, durante três meses. Ao final do tratamento, a evolução das intensidades de dor de cada paciente foi avaliada pelo Cardiff Breast Score, para determinação da responsividade terapêutica. A taxa de resposta terapêutica foi 71,4% e 76,4%, respectivamente para o óleo de borragem e de girassol, não havendo diferença, estatisticamente significativa, entre os medicamentos. A cefaléia, o estresse percebido e o edema mamário reduziram-se em ambos os grupos. Três mulheres do grupo do óleo de girassol apresentaram sangramento vaginal profuso. Concluiu-se que ambos os medicamentos foram eficazes no tratamento da mastalgia cíclica moderada à intensa. São necessários estudos adicionais para determinar a relação custo/benefício do óleo de girassol.

Cyclical mastalgia is a frequent condition and represents an exacerbation of the physiological process, recurring in the luteal phase of the menstrual cycle. When it is intense, it impairs a woman's quality of life, because if interferes on her daily activities. In order to make a comparative evaluation of the effectiveness and adverse effects of borage and sunflower oils in the treatment of moderate to severe cyclical mastalgia, a double-blind randomized study of clinical trial type was carried out between February 2001 and December 2002 involving 111 women. The intensity of breast pain was assessed on a monthly basis by means of the analogic visual scale. The participants were subdivided as follows into two groups according to the drug administered: 56 were given borage oil tablets and 55 sunflower oil, 1 tablet per day for both groups over a period of three months. At the end of the treatment, the evolution in the intensity of the pain of each patient was compared with the Cardiff Breast Score for an assessment of the responsiveness to treatment. The responsiveness rates were 71.4% and 76.4%, respectively for the borage and sunflower oil groups, no statistically significant differences being found. Although headache, perceived stress and mammary edema had reduzed in both groups; uterine bleeding occurred in three patients, as a serious side effect of sunflower oil. Further studies, however, are needed to determine the cost-benefit ratio of sunflower oil.

O tratamento da fístula mamária através da técnica da incisäo transareolopapilar / The treatment of mammary fistula by means of transareolopappilary incision

Os autores revisaram a literatura quanto ao tratamento cirúrgico utilizado para fístula mamária, observando índice de recidiva significativamente elevado. Descrevem técnica cirúrgica pessoal que resultou em baixo índice de recidiva e bom resultado estético. A fístula mamária foi descrita por Zuska e cols. (1951), porém Atkins (1953) descreveu com detalhes a sua apresentaçäo e tratamento. Trata-se de doença de evoluçäo crônica näo relacionada à funçäo d lactaçäo e que ocorre com mais frequência entre 30-40 anos