[Nutritional and Functional assessment of peritoneal dialysis patients in the clinical practice: Report from MITO-DP Group]. / Valutazione Nutrizionale e Funzionale dei pazienti in dialisi peritoneale nella pratica clinica: l'esperienza del Gruppo Medico-Infermieristico Toscano di Dialisi Peritoneale (M.I.TO.-DP).

Nutritional abnormalities and physical inactivity are risk factors of increased morbidity and mortality in patients with ESRD. Identify and define malnutrition, in particular protein-energy depletion (PEW), is an important task in the management of renal patients. The aim of this multicenter observational study was to implement the assessment of nutritional status and functional capacity in patients on peritoneal dialysis, including tests and validated methods which are relatively easy to apply in daily clinical practice. The study includes all the 133 prevalent patients (80 m, 53 f, age 65 14 years), in peritoneal dialysis treatment (vintage 26 19 months) in 9 centers in Tuscany. We performed anthropometry, bioimpedance (BIA), clinical biochemistry, evaluation of habitual physical activity (RAPA tests) and performance (Sit-To-Stand test), appetite-evaluation questionnaire, and indices including the Malnutrition Inflammation Score (MIS), Geriatric Nutrition Risk Index (GNRI), Charlson comorbidity index, Barthel and Karnowsky index. The latter showed a condition of dependence in 7.2% and 19.7% of cases, respectively. Poor appetite was recorded in 48.2%. The majority of patients fell within the overweight / obesity range (51%) with waist circumference values associated with increased cardiovascular risk in 51% of males and 60% of females. At the BIA analysis, a BCMI <8 kg/m2 was detected in 39% of patients; an estimated protein intake <1.0 g / kg/d was found in 59% of cases; 34% of patients had serum albumin <3.5 g / dl; control of acidosis was good (bicarbonate 25.4 3.8 mM) but hyperphosphatemia was present in 64.6% of patients. A condition of sedentary or light physical activity was reported by 65.1% of patients, vigorous activity only by 11.9%. The 86.5% of patients able to perform the Sit-to-stand test reported a lower than the reference values for age and sex. A diagnosis of PEW was possible in 8% of our series, while a MIS score> 11, indicative of PEW, took place in 12.7% of cases. The values of the MIS correlated directly with age and the degree of comorbidity and inversely with the sit-to-stand test, RAPA tests and appetite level. The data in this study show that single tests indicative of malnutrition disorders are frequent to be found in our series of peritoneal dialysis patients. However, a diagnosis of PEW is quite infrequent. A large percentage of patients are overweight with increased abdominal adiposity, and reduced cell mass and protein intake below recommended levels; the level of habitual physical activity is low, and the level of physical capability is scarce. Therefore it is conceivable a nutritional counseling intervention to increase the intake of proteins, limiting the phosphorus and (when indicated) energy intake and to stimulating spontaneous physical activity or arranging assisted programs for functional rehabilitation. Close monitoring of the nutritional status and implementation of programs of adapted physical activity should have a prominent role in the clinical management of patients on peritoneal dialysis.

Bone integration of new stemless hip implants (proxima vs. nanos). A DXA study: preliminary results.

The development of short femoral prostheses has the advantage to preserve bone and soft tissues, restore hip geometry, permit mini-invasive techniques and allow quickly return to an active life, but very few studies described bone reaction to these new designed prostheses. The aim of the present study was to evaluate the osseointegration of two different partial neck retained stemless hip prosthesis at one year after surgery, measured by the changes of periprosthetic bone mineral density (BMD) in 5 regions of interest (ROIs) using a dual-energy X ray absorptiometry (DXA) device. The signs of stress-shielding were evaluated by standard radiographs. Thirty-two uncemented primary total hip arthroplasty (THA) patients allocated into 2 groups were evaluated. In the first group (n=19) a Proxima (De-Puy-J&J) hip stem was implanted. In the second group (n=12) a Nanos (Smith & Nephew) hip stem was used. We found that both the implants preserve metaphyseal bone stock and increase periprosthetic BMD. In Nanos prostheses a significant higher BMD values were observed in region of interest (ROI) 3 and 4 (p<0.05). No differences were found in ROIs 1, 2, and 5. Proxima stem seem to produce a physiological strain distribution in the femur. No signs of stress-shielding were present in both the implants. In conclusion, this preliminary DXA analysis showed a physiological integration of both the stems that reproduces the biomechanical stress of proximal femur. New designed short stem implants showed optimal osseointegration after one year, and therefore appears an excellent alternative to traditional long stem hip prostheses.

Rhizosphere dynamics during phytoremediation of olive mill wastewater.

The potential of phytoremediation as a treatment option for olive mill wastewater (OMW) was tested on five perennial tree species. Cupressus sempervirens and Quercus ilex proved tolerant to six-month OMW treatment followed by six-month water irrigation, whereas Salix sp. and Laurus nobilis and, later, Pinus mugo suffered from phytotoxic effects. Test plants were compared to controls after treatment and irrigation, by monitoring biochemical and microbiological variations in the rhizosphere soil. OMW-treated soils were exposed to 50-fold higher phenols concentrations, which, irrespective of whether the respective plants were OMW-resistant or susceptible, were reduced by more than 90% by the end of the irrigation cycle, owing to significantly increased laccase, peroxidase and ß-glucosidase activities, recovery/acquisition of bacterial culturability and transitory development of specialized fungal communities sharing the presence of Geotrichum candidum. Of all results, the identification of Penicillium chrysogenum and Penicillium aurantiogriseum as dominant rhizosphere fungi was distinctive of OMW-tolerant species.

Mid-term results of a custom-made short proximal loading femoral component.

Bone preservation and physiological distribution of forces on the proximal femur are key elements in introducing a successful uncemented total hip replacement. In order to achieve this, in the mid 1990s, we developed an ultra short proximal loading custom-made component with a lateral flare, a high femoral neck osteotomy and without a diaphyseal stem. We report the outcome of 129 custom-made hydroxyapatite-coated uncemented short femoral components inserted into 109 patients between June 1995 and May 2004. The mean age of the patients was 51 years (21 to 71) and the mean follow-up was eight years (4.9 to 14.1). Bone behaviour around the implant was studied on the post-operative radiographs. The mean Harris hip score improved from 44 (8 to 66) pre-operatively to 95 (76 to 100) at final follow-up. The Western Ontario MacMaster University Osteoarthritis index was 93 of 100 at final review. None of the patients reported thigh pain. A total of five hips were revised, three for polyethylene liner exchange and two for complete revision of the acetabular component. No femoral components were revised. The radiological changes in the proximal femur were generally good, as evidenced by spot welds both on the medial and lateral aspects of the femur. No component migrated. The presence of a lateral flare and use of a high osteotomy of the femoral neck provided good clinical and radiological results. The absence of a diaphyseal portion of the stem did not impair stability.

Quorum sensing inhibition activity of garlic extract and p-coumaric acid.

AIMS: The goal of this work was to investigate the influence of DMSO, garlic extract and p-coumaric acid on bacterial quorum sensing (QS). METHODS AND RESULTS: The decreases in the QS responses of QS reporter strains Escherichia coli pSB401 and pSB536, Agrobacterium tumefaciens NTL4, Chromobacterium violaceum 5999 and wt 494, Pseudomonas putida IsoF/gfp and environmental Pseudomonas chlororaphis were quantified in relation to growth inhibitory effects. DMSO showed no significant QS-specific effects on the strains tested even at close-to-lethal concentrations. Garlic extracts antagonized the activity of QS receptors LuxR, AhyR and TraR, but were toxic at higher concentrations. P-coumaric acid fully inhibited QS responses of 5999, NTL4 and P. chlororaphis, with no influence on cell viability. CONCLUSIONS: The quorum sensing inhibition activity of garlic was extended to novel receptors, and p-coumaric acid was found to possess previously undescribed QS antagonist properties. SIGNIFICANCE AND IMPACT OF THE STUDY: The results suggest that p-coumaric acid might act as QS inhibitor. Further studies are required to understand its role in the regulation of QS and investigate structurally related compounds.

Bone remodelling in THA: A comparative DXA scan study between conventional implants and a new stemless femoral component. A preliminary report.

Periprosthetic bone loss is a major cause for concern in patients undergoing total hip arthroplasty (THA). There are many different factors that may determine the pattern of bone loss and bone remodelling following THA, such as the quality of the bone before the hip replacement, skeletal bone mass at the time of the operation, material and method of fixation and implant design. Recent developments in dual-energy X-ray absorptiometry (DXA) have made it possible to quantify bone mineral density (BMD) to evaluate changes around the prosthesis and to measure bone stock and bone density redistribution after a total hip replacement. In this cross-sectional multicentre clinical study the DXA method was used to compare bone mass after uncemented THA of a custom-made stemless design with five groups of conventional cementless implants (Alloclassic, Mayo, CFP, IPS, ABG). The adaptive bone changes of the proximal femur three years after implantation were evaluated. Periprosthetic BMD was measured in 130 subjects in the seven regions of interest (ROI) based on Gruen zones. Significant differences were found between the stemless implant and the other five groups in zones 1, 4 and 7. The CFP, IPS, and ABG groups showed decreased BMD in ROI 1, and the Mayo, IPS and Alloclassic in ROI 7. An increased BMD in ROI 4 was observed in the Mayo, IPS, ABG and Alloclassic groups. The results of the present study suggest that a conservative stemless implant with complete proximal load transfer produces a homogeneous and more physiological redistribution of bone density, allowing maintenance of proximal periprosthetic bone stock.

Bone preservation with a conservative metaphyseal loading implant.

Proximal load transfer and the absence of distal fixation of the stem are crucial to obtain the best behaviour of the femoral bone after total hip replacement. In this study, dual energy X-ray absorptiometry (DEXA) was employed to understand and compare the bone density changes and thus the re-distribution of mechanical forces in two different extra short stems. Two cohorts of ten patients were included in this retrospective study. Both implants are custom-made and present a well defined lateral flare. The first model is fully coated and presents a short stem (Group A), the second is an unstemmed metaphyseal implant with a polished tip (Group B). DEXA scans were obtained in all patients at the two-year follow-up. A higher BMD was detected in ROIs 1, 2, 4, 5 in Group B confirming a preservation of the proximal bone mass and thus indirectly a more proximal load transfer. The results obtained confirm the importance of the geometry of the implant on proximal bone density. Loading both medial and lateral proximal femoral flares and the complete absence of the diaphyseal portion of the stem provide the optimal bone remodelling of the femur after total hip replacement.

Ultra-short stems with proximal load transfer: Clinical and radiographic results at five-year follow-up.

Bone stock preservation is crucial when performing THR in young patients. Following this philosophy, an original ultra-short custom-made implant with extensive proximal load transfer was developed. It presents three very innovative features: absence of the diaphyseal portion of the stem, a well defined lateral flare with load transfer on the lateral column of the femur and a very high femoral neck cut. These innovations resulted in a very conservative implant both of the bone stock and the soft tissues. In this study we reviewed the X-rays of 111 patients with 131 primary total hip replacements performed with this implant. The average pain score using the Harris Hip Score system, at an average of five years after surgery, was 42 of 44 points; 95 per cent (124) of the patients had no or slight pain. We had no cases of thigh pain. None of the patients required a femoral stem revision. Two cases had to be re-operated for polyethylene liner exchange but the femoral implant was stable and left in place. At five years, all implants appeared radiographically stable with well maintained proximal bone stock. It was concluded that the geometry of this implant provides significant initial stability, which seems to be preserved throughout a long follow-up period. This study validates the assumption that torsional loads can be controlled even without the diaphyseal portion of the stem and that neck preservation combined with lateral flare support guarantees a more natural loading of the femur.

Isocratic separation of monosaccharides using immobilized Concanavalin A.

The successful fractionation of a mixture of monosaccharides using an immobilized lectin column operating under isocratic conditions is described. The key factors for effective separation were found to be sample size, feed velocity and column length. Under optimal conditions it was possible to obtain complete resolution of a mixture of L-arabinose, D-fructose and D-mannose in less than 40 min.

Shoulder prosthesis in reconstruction of the scapulohumeral girdle after wide resection to treat malignant neoformation of the proximal humerus.

Progress in preoperative chemotherapy and radiation therapy, both intra- and postoperative, has in time allowed for an increase in indications for shoulder implant surgery in malignant tumors, thus drastically reducing the number of amputations. The use of prostheses, particularly those of the more recent generation, respond to the needs to overcome the limits of loss of movement, as long as good anatomical reconstruction of the soft tissues, the premise for good functional and cosmetic recovery, is also possible. The current tendency is that of using modular shoulder prostheses that provide a wide variety of sizes. This type of prosthesis has surpassed custom made prostheses in terms of simplicity, adaptability and economy. The use of cementation in stabilization of the intramedullary stem has made them more stable even in the long term, allowing for early rehabilitation. The main problem in reconstruction with a prosthesis is the quantity of residual muscular tissue (deltoideus, extrarotators) and the stabilization system of the same to the prosthesis in order to avoid dislocation, which constitutes the main complication. It is the purpose of this study to present the clinical and functional results obtained in 23 implants carried out for primary malignant neoformation of the upper limb. The implants studied were of three types (custom made, modular in Cr-Co-Mb and Ti-Al-Va). The results reported show how a shoulder prosthesis can obtain good functional results, particularly when a part of the activator muscles of the shoulder such as the deltoideus and the extrarotators is preserved. When the contrary is true, the results obtained are modest because the prosthesis acts as a simple spacer.

Cutting edge: positive selection induced by a self-peptide with TCR antagonist activity.

Antagonist-like engagement of the TCR has been proposed to induce T cell selection in the thymus. However, no natural TCR ligand with TCR antagonist activity is presently known. Using a combination of bioinformatics and functional testing we identified the first self-peptide that can both deliver antagonist-like signals and promote T cell selection in the thymus. The peptide is presented by appropriate MHC class I molecules in vivo. Thus, endogenous antagonist peptides exist and may be involved in TCR repertoire selection.

Peptide loading of nascent MHC class I molecules.

A critical molecular interaction during assembly of the major histocompatibility complex (MHC) class I molecules takes place between the heavy chain and the transporter-associated with antigen-processing (TAP) complex. The recent mapping of regions of the heavy chain involved in the binding to TAP suggests a complex molecular interaction essential for the cell surface expression of the MHC class I. The advances made in understanding the TAP-MHC class I interaction are reviewed and discussed here.

Modulation of CD8+ T cell response to antigen by the levels of self MHC class I.

The response of H-Y-specific TCR-transgenic CD8(+) T cells to Ag is characterized by poor proliferation, cytolytic activity, and IFN-gamma secretion. IFN-gamma secretion, but not cytotoxic function, can be rescued by the B7.1 molecule, suggesting that costimulation can selectively enhance some, but not all, effector CD8(+) T cell responses. Although the H-Y epitope binds H-2D(b) relatively less well than some other epitopes, it can induce potent CTL responses in nontransgenic mice, suggesting that the observed poor responsiveness of transgenic CD8(+) T cells cannot be ascribed to the epitope itself. Previously reported reactivity of this TCR to H-2A(b) is also not the cause of the poor responsiveness of the H-Y-specific CD8(+) T cells, as H-Y-specific CD8(+) T cells obtained from genetic backgrounds lacking H-2A(b) also responded poorly. Rather, reducing the levels of H-2(b) class I molecules by breeding the mice to (C57BL/6 x B10.D2)F(1) or TAP1(+/-) backgrounds partially restored cytotoxic activity and enhanced proliferative responses. These findings demonstrate that the self MHC class I gene dosage may regulate the extent of CD8(+) T cell responsiveness to Ag.

Results of the anatomic cementless prosthesis with different types of hydroxyapatite coating.

This study evaluates the clinical and radiographic results and the bone-implant osteointegration obtained with the Anatomic (Zimmer, Warsaw, Ind) cementless total hip prosthesis using three different types of surface coating. Two hundred twenty-seven patients underwent total hip arthroplasty using the Anatomic prosthesis. Patients were divided into groups based on the type of surface coating: in group A (69 patients), prostheses were uncoated; in group B (90 patients), the metaphyseal region of the prostheses was coated with calcicoat (a mixture of hydroxyapatite and tricalcium phosphate) (Zimmer); and in group C (68 patients), the fiber mesh and proximal stem of the prostheses were coated with calcicoat. Prostheses coated with calcicoat yielded better clinical and radiographic results than uncoated prostheses, especially in regard to thigh pain. No significant difference was observed between groups B and C. These results obtained with the Anatomic prosthesis are promising and encourage the use of implants coated with calcicoat.

Alpha beta TCR+ cells are a minimal fraction of peripheral CD8+ pool in MHC class I-deficient mice.

MHC class I molecules play a role in the maintenance of the naive peripheral CD8+ T cell pool. The mechanisms of the peripheral maintenance and the life span of residual CD8+ cells present in the periphery of beta 2-microglobulin-deficient (beta 2m-/-) mice are unknown. We here show that very few CD8+ cells in beta 2m-/- mice coexpress CD8 beta, a marker of the thymus-derived CD8+ T cells. Most of the CD8 alpha+ cells express CD11c and can be found in beta 2m/RAG-2 double-deficient mice, demonstrating that these cells do not require rearranged Ag receptors for differentiation and survival and may be of dendritic cell lineage. Rare CD8 alpha+CD8 beta+ cells can be detected following in vivo alloantigenic stimulation 2 wk after the adult thymectomy. Selective MHC class I expression by bone marrow-derived cells does not lead to an accumulation of CD8 beta+ cells in beta 2m-/- mice. These findings demonstrate that 1) thymic export of CD8+ T cells in beta 2m-/- mice is reduced more severely than previously thought; 2) non-T cells expressing CD8 alpha become prominent when CD8+ T cells are virtually absent; 3) at least some beta 2m-/- CD8+ T cells have a life span in the periphery comparable to wild-type CD8+ cells; and 4) similar ligands induce positive selection in the thymus and survival of CD8+ T cells in the periphery.

Molecular analysis of UAS(E), a cis element containing stress response elements responsible for ethanol induction of the KlADH4 gene of Kluyveromyces lactis.

KlADH4 is a gene of Kluyveromyces lactis encoding a mitochondrial alcohol dehydrogenase activity, which is specifically induced by ethanol and insensitive to glucose repression. In this work, we report the molecular analysis of UAS(E), an element of the KlADH4 promoter which is essential for the induction of KlADH4 in the presence of ethanol. UAS(E) contains five stress response elements (STREs), which have been found in many genes of Saccharomyces cerevisiae involved in the response of cells to conditions of stress. Whereas KlADH4 is not responsive to stress conditions, the STREs present in UAS(E) seem to play a key role in the induction of the gene by ethanol, a situation that has not been observed in the related yeast S. cerevisiae. Gel retardation experiments showed that STREs in the KlADH4 promoter can bind factor(s) under non-inducing conditions. Moreover, we observed that the RAP1 binding site present in UAS(E) binds KlRap1p.

Species-specific selection on soil fungal population after olive mill waste-water treatment.

Soil was treated with olive mill waste water (OMW) in order to study the effect of this agriculture waste on soil fungal population. Changes in fungal composition were observed after soil pollution. In order to test OMW selective pressure, growth kinetics of Penicillium cyclopium, Scopulariopsis brevicaulis and Cladosporium cladosporioides were studied on solid media supplemented with different OMW concentrations. S. brevicaulis and C. cladosporioides did not grow at OMW concentration higher than 50%, while at concentrations lower than 50% a growth decrease was observed. Instead, P. cyclopium was able to actively grow at all concentrations of OMW tested. Therefore the OMW can influence and modify the soil fungal homeostasis.

Identification of a domain of Trypanosoma cruzi metacyclic trypomastigote surface molecule gp82 required for attachment and invasion of mammalian cells.

Recombinant proteins and synthetic peptides representing various sequences of gp82, a surface glycoprotein of Trypanosoma cruzi metacyclic trypomastigotes implicated in mammalian cell invasion, were used in this study aiming at the identification of the domain(s) of this molecule required for interaction with target cells. Invasion of cultured HeLa cells by metacyclic trypomastigotes was inhibited by about 80% in the presence of native gp82 or the corresponding recombinant construct J18. Inhibition by recombinant proteins J18a and J18b, containing respectively the N-terminal and the C-terminal portions of gp82, was on the order of 30% and 65%. As compared to J18b (amino acids 224-516), the truncated gp82 fragments J18b1 (amino acids 303-516) and J18b2 (amino acids 357-516) displayed lower inhibitory effect (approximately 40% and approximately 15%, respectively). Compatible with these observations, we found that the recombinant protein J18b, but not J18a or J18b2, binds to HeLa cells in a dose-dependent and saturable fashion. Experiments with ten overlapping synthetic peptides, representing the gp82 portion spanning amino acids 224-333, showed that peptides 4 (amino acids 254-273) and 8 (amino acids 294-313) have significant inhibitory activity on HeLa cell invasion by metacyclic forms. All these results indicate that the portion of gp82 required for mammalian cell attachment and invasion is located in the central domain of the molecule.

A recombinant protein based on the Trypanosoma cruzi metacyclic trypomastigote 82-kilodalton antigen that induces and effective immune response to acute infection.

To further investigate the immunological properties of the stage-specific 82-kDa glycoprotein (gp82) of Trypanosoma cruzi metacyclic trypomastigotes, previously shown to induce antigen-specific humoral and T-cell responses in mice, we performed a series of experiments with recombinant proteins containing sequences of gp82 fused to glutathione S-transferase. Of five fusion proteins tested, only J18b and J18b1, the carboxyproximal peptides containing amino acids 224 to 516 and 303 to 516, respectively, were recognized by monoclonal antibody 3F6 as well as by various anti-T. cruzi antisera and, when administered to mice, were capable of eliciting antibodies directed to the native gp82. The amino-terminal peptide and other carboxyterminal recombinant proteins lacking the central domain of gp82 (amino acids 224 to 356), which is exposed on the surface of live metacyclic forms, did not display any of these properties. Spleen cells derived from mice immunized with any of the five recombinant proteins proliferated in vitro in the presence of native gp82.J18b was the most stimulatory, whereas J18b3, the peptide containing amino acids 408 to 516, elicited the weakest response. When BALB/c mice immunized with J18b antigen plus A1(OH)3 as adjuvant were challenged 10 5 metacyclic trypomastigotes, 85% of them resisted acute infection, in comparison with control mice that received glutathione S-transferase plus adjuvant. Antibodies induced by J18b protein lacked agglutinating or complement-dependent lytic activity and failed to neutralize parasite infectivity. On the other hand, CD4+T cells from the spleens of J18b-immunized mice displayed an intense proliferative activity upon stimulation with 1.25 microgram of native gp82 per ml, which resulted in increased production of gamma interferon, a cytokine associated with resistance to T. cruzi infection.

[Clinical usefulness of a computerized tomography study of morphology of the femoral canal in patients candidates for hip prosthesis]. / Utilità clinica dello studio con tomographia computerizzata della morfologia del canale femorale nei pazienti candidati a protesizzazione dell'anca.

In the last few years hip arthroplasty has been increasingly used and both metaphysis and diaphysis of the proximal femur are studied preoperatively for best compliance between prosthesis and bone. Indeed, the best results can be obtained by reducing the risk of stem end mobilization, which means to choose the prosthesis fitting the femoral canal best and to limit the use of cement prostheses, which are at high risk of mobilization in time, to advanced osteoporosis patients. We used a simple and repeatable CT technique to study femoral canal structure and size. After accurately positioning the patients inside the gantry, we acquired some axial scans at scheduled levels referring to the horizontal midline of the lesser trochanter. The axial scans were acquired 2 cm above and 5 and 10 cm below the horizontal midline in 105 patients. Measurements were bilateral in 13 patients. Finally, the results were compared with the surgical outcome. Femoral canal cross-diameters and cortical bone width varied greatly, which variations were confirmed in bilateral measurements and in the same patient between the two femurs. Femoral canal structure, which can be remodelled, influences the choice of the stem and thus the amount of cement. We believe this method to be fundamental for correct surgical planning and for best treatment outcome.