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The thermal diffuse scattering (TDS) in X-ray diffraction (XRD) patterns contains significant information about the local lattice dynamic structure of nanocrystalline systems. Techniques such as the pair distribution function (PDF) are commonly employed to extract this information, where the correlated movement of atomic pairs remains encoded in the breadths of the PDF peaks. However, PDF techniques require a Fourier transformation of the experimental XRD data, orientationally averaging the local dynamic information, rendering it not readily distinguishable from the static component and crystallite size and shape effects. Herein, we explore the possibility of an analysis of local lattice dynamics based directly on XRD powder pattern modeling, where TDS is added to the structural model of the traditional Rietveld method. Allied with the whole powder pattern modeling approach, the crystallite shape and static components are simultaneously estimated. Two study cases of Pd nanocrystalline systems are analyzed: (i) in silico nanosphere powder simulations via molecular dynamics (MD) and (ii) synchrotron radiation XRD powder patterns of Pd nanocubes. In silico analysis points out that the TDS model provides the correct trends of the correlated atomic movement up to the ninth coordination shell. The experimental case shows that this TDS model correctly estimates the force mechanisms of the nanocrystalline Pd system. We believe that the established method and the obtained results in this study broaden the application scope of XRD for studying the dynamic properties of nanocrystalline materials.

Acute mastoiditis (AM) is the most common complication of acute otitis media and primarily affects children under the age of two; current data on its prevalence in paediatric patients with cochlear implant (CI) are still scant. Proper management of AM in CI children is crucial in order to avoid the implications (financial and emotional) of an explant.

Background: : Chronic respiratory diseases (CRD) are a major public health problem worldwide. In the current epidemiological context, CRD have received much interest when considering their correlation with greater susceptibility to SARS-Cov-2 and severe disease (COVID-19). Increasingly more studies have investigated pathophysiological interactions between CRD and COVID-19. Area covered: : Animal experimentation has decisively contributed to advancing our knowledge of CRD. Considering the increase in ethical restrictions in animal experimentation, researchers must focus on new experimental alternatives. Two-dimensional (2D) cell cultures have complemented animal models and significantly contributed to advancing research in the life sciences. However, 2D cell cultures have several limitations in studies of cellular interactions. Three-dimensional (3D) cell cultures represent a new and robust platform for studying complex biological processes and are a promising alternative in regenerative and translational medicine. Expert opinion: : Three-dimensional cell cultures are obtained by combining several types of cells in integrated and self-organized systems in a 3D structure. These 3D cell culture systems represent an efficient methodological approach in studies of pathophysiology and lung therapy. More recently, complex 3D culture systems, such as lung-on-a-chip, seek to mimic the physiology of a lung in vivothrough a microsystem that simulates alveolar-capillary interactions and exposure to air. The present review introduces and discusses 3D lung cultures as robust platforms for studies of the pathophysiology of CRD and COVID-19 and the mechanisms that underlie interactions between CRD and COVID-19.

ABSTRACTThis work aimed the production of eugenyl acetate through heterogeneous chemical catalysis of eugenol and acetic anhydride as substrates by commercial Lewatit ® GF 101 resin as catalyst. From the experimental design, the highest reaction conversion (94.85%) was obtained after 10 min of reaction time (1:1 substrates molar ratio, 1 wt% of catalyst content at 70°C), and up to 45 min a complete conversion was reached. The catalyst reuse was investigated, and after 10 reuse cycles, no loss of catalytic activity was observed. Antimicrobial activity of eugenyl acetate against Gram-positive (Staphylococcus aureus ATCC 9763, and Listeria monocytogenes ATCC 15117) and Gram-negative bacteria (Escherichia coli ATCC 25922 and Pseudomonas aeruginosa ATCC 27853) showed higher inhibition zone against to L. monocytogenes (42 mm). Besides, the eugenyl acetate antibacterial effect was also demonstrated by the minimum inhibitory concentration, and P. aeruginosa had higher inhibition (41.77 mm) at lower eugenyl acetate concentration (2500 μg.mL−1).

AbstractTwo experiments were conducted to investigate the inclusion of high cut whole plant corn silage (HCCS) in diets for finishing heavy pigs on digestibility, metabolic utilisation, growth performance and slaughter traits. A control diet (CTR, containing maize meal, barley meal, extracted soybean meal and wheat bran, 550, 250, 90 and 80 g/kg DM, respectively) was compared with diet (CS) containing 200 g/kg DM of HCCS. The HCCS replaced wheat bran and part of maize meal in the CTR diet. In the first experiment, eight barrows were used in a two periods cross over design with periods of 21 d, included 7 d of total collection and three cycles of 24 h each in a respiratory chamber. In the second experiment, 28 barrows were divided into pairs on the basis of BW, kept in 14 pens and fed with the experimental diets until slaughter. Lower DM, OM, CP and energy total tract apparent digestibility was measured for the CS diet. Overall P retention as percentage of P intake was higher for CS diet, while N retention was similar for the two diets. Pigs fed CS tended to have a lower retained energy and the estimated NE of maize silage was 8.47 MJ/kg DM. Pigs fed the CS diets had a lower daily gain, a lower BW at slaughter and a reduction in the weight of back fat. The inclusion of HCCS increased the size of the stomach, the aNDFom concentration of stomach content and reduced the incidence of follicular gastritis.

Sinonasal polyposis (SNP) is a chronic inflammatory disease of nasal and paranasal cavities. Human leukocyte antigen-G molecules (HLA-G) are non-classic HLA-I molecules with anti-inflammatory and tolerogenic properties. HLA-G production is mainly induced by interleukin (IL)-10. IL-10 is an anti-inflammatory cytokine that inhibits the production of proinflammatory cytokines and induces HLA-class II down-modulation. Recent studies suggest that HLA-G could play a role in SNP pathogenesis; in SNP patients physiological levels of IL-10 (produced by activated peripheral blood CD14+ monocytes) are not able to induce production of HLA-G. Different mechanisms could justify these findings: genomic or amino-acidic sequence alterations in IL-10 lower IL-10 receptor expression, lower IL-10 receptor affinity, or alterations of the intracellular signal transmission. This study analyzes nucleotidic sequence of IL-10 gene in SNP patients. Sequencing of IL-10 gene shows that the lack of HLA-G production by peripheral blood CD14+ monocytes is not related to alterations in IL-10 gene nucleotidic sequence.

The aim of this work was to evaluate the effects of the inclusion of whole-ear corn silage (WECS) in diets for advanced fattening heavy pigs (substitution for part of the dry corn and wheat bran) allocated or not in metabolic cages on the main blood parameters. The high-moisture shelled corn is largely used in pig feeding while WECS is less often used despite the fact that it increases the DM crop yield. Three experimental diets were fed to 27 barrows (Italian Large White × Italian Duroc), with an average BW of 98.2 (±5.6) kg at the start of the trial, and randomly allotted to 3 experimental groups including a control diet (CON) containing cereal meals (corn, barley, and wheat, 80.2% DM in total), soybean meal (9% DM), wheat bran (8% DM), minerals and supplements (2.8% DM), and 2 diets containing WECS (15 or 30% DM referred to as 15WECS and 30WECS, respectively) in partial or complete substitution for wheat bran and corn meal. The pigs were randomly housed in 9 pens with 3 animals per pen and 3 pens per dietary treatment. Six pigs per each of the 3 treatments were moved from the pens to individual metabolic cages for 3 consecutive periods (2 pigs per treatment per period). Each period lasted 14 d, and blood was collected at the start and at the end of the periods. Blood was drawn from the jugular vein before feed distribution in the morning, at 14 d intervals, and analyzed for hematological, metabolic, and serum protein profiles. The effect of the metabolic cage housing was included in the statistical model to compare the results obtained in the 2 different environments of restrained and group-housed barrows. The WECS affected the neutrophil to lymphocyte ratio and mean corpuscular hemoglobin concentration. The main diet effect on plasma metabolites was recorded for plasma NEFA, with higher values in WECS diets compared with the CON. The metabolic cage housing affected both hematological (red blood cell count, hemoglobin, hematocrit) and metabolic (protein and its fractions) items, which can be markers of hemodilution. These results indicate the possibility to use this feed in the diet of heavy pigs without negative effects on physiology. The absolute values from metabolic profile of pigs in metabolic cages must be considered with caution for possible comparisons with values obtained on-field in group pens, particularly because a different hemodilution may affect the results.

Los métodos geofísicos constituyen una herramienta ampliamente utilizada en exploración mineral. Este trabajo presenta y discute los resultados de estudios geológicos y geofísicos desarrollados en un yacimiento de manganeso de origen supergénico, localizado en la región sudeste de Brasil. La zona mineralizada descrita en levantamientos geológicos fue caracterizada por bajos valores de resistividad (20Ω.m) y altos valores de cargabilidad (30ms), en un patrón similar al descrito en diversos trabajos en depósitos minerales de óxidos y sulfuros en rocas. Modelos geofísicos pseudo-3D permita la generación de mapas para diversas profundidades. Las áreas de alta cargabilidad y baja resistividad definen un patrón de mineralización gonditica con altos niveles de Mn. Áreas considerables con elevados valores de cargabilidad y baja resistividad probablemente resulten de la acumulación de hidróxido de manganeso y hierro, originados del intemperismo en cuerpos de mineral gonditico, disolución, percolación y precipitación.

The aim was to evaluate 2 levels of dietary inclusion of chopped whole-ear corn silage (WECS) on energy and nutrient utilization, growth, and slaughter performances of heavy pigs. Two in vivo experiments were conducted to determine digestibility and metabolic utilization of WECS using 18 barrows weighing 118 ± 8 kg BW on average, metabolic cages and respiration chambers (Exp. 1), and the effect of WECS on the growth performance and carcass traits on 42 barrows from 90 to 170 kg BW (Exp. 2). In both experiments, pigs were fed 3 experimental diets: a control diet (CON) containing cereal meals, extracted soybean meal, and wheat bran (80%, 9%, and 8% of DM, respectively) and 2 diets containing 15% (15WECS) or 30% WECS (30WECS) on a DM basis in place of wheat bran and corn meal. The diets were prepared daily by mixing the WECS to a suitable compound feed. Feed intake was always restricted to allow a daily DMI of 7.2% BW0.75in Exp. 1 and from 8.0% to 6.5% BW0.75in Exp. 2. Diets had similar NDF contents (15.2% to 15.8% of DM), and WECS inclusion resulted in a slight reduction in CP content (from 14.0% to 13.6% of DM) and a considerable decrease in P content (from 0.47% to 0.30% of DM). Digestibility of OM, CP, and fat was similar among diets, whereas P digestibility was lower (P< 0.05) for the 30WECS diet (33.5%) in comparison with the CON and 15WECS diets (45.5% and 44.1%, respectively). Nitrogen lost in feces and urine and N retained were not different among diets, whereas P retained decreased with the increase of WECS (5.4, 3.7, and 2.2 g/d for the CON, 15WECS, and 30WECS diets, respectively; P< 0.05). No difference among diets was observed for energy balance. The WECS contained 13.48 MJ ME and 9.39 MJ NE/kg DM. In Exp. 2, feed intake was not depressed by WECS inclusion, and the ADG for the whole experiment was not different among dietary treatments (from 737 to 774 g/d). Fecal pH was lower (P< 0.05) for the WECS diets than the control diet (7.10 and 7.00 vs. 7.40) and for the sampling at 150 kg BW than that at 130 and 110 kg BW (6.96 vs. 7.29 and 7.24). At slaughter, lean percentage in the carcass was lower in the 30WECS diet than those of the other 2 diets (46.8% vs. 48.3% and 48.6%, P= 0.05). The overall experimental data obtained in both trials indicate that substitution of wheat bran and corn meal for WECS (up to 30% of DM) does not affect, with the exception of P utilization and carcass leanness, energy and nutrient utilization and performance of heavy pigs in the last phase of growing.

Fine-needle aspiration cytology (FNAC) is a minimally invasive procedure usually well tolerated, easy to perform, quick, cheap and easy to repeat in case of doubts or non-diagnostic results. Echography is also a fast, cheap and non-invasive tool; however, the role of FNAC and echography in the diagnosis of salivary gland pathology is not universally recognised. Three hundred and fifty-seven patients with a cytological diagnosis at FNAC, and 247 of these who were also studied with echography, were enrolled for this retrospective study. The final histopathological diagnoses, obtained after surgery, were then compared to the preoperative FNAC diagnoses and echographic findings. From the analysis of our data, the overall FNAC specificity resulted 93%, sensitivity 83%, and diagnostic accuracy 92%. Echography sensibility was 57.1% specificity 98.2%, while positive and negative predictive value were respectively 80% and 94.8%. While echography can be useful in order to provide a better characterization of salivary gland lesions, FNAC can then be considered a safe diagnostic tool with reliable sensitivity and specificity for the assessment of salivary gland pathology and thus for selecting patients and indicating the best surgical treatment.

Automatic administration of drugs to control cardiovascular function during and after surgery has received considerable attention. Although the potential benefits associated with such a technology remain unquestioned, the several adaptive control strategies proposed thus far have had very limited success in practice. In developing robust adaptive control methodologies for drug dosing in cardiovascular applications, we have analysed a well-known multiple-model adaptive control strategy for blood pressure control. The results reveal that no guarantee of protection against actually inserting a destabilising controller into the closed-loop is given and one cannot even put a global upper bound on the time during which the destabilising controller is attached. We advocate caution towards issues which in the past may have been either disregarded or not subjected to a systematic analysis as instability could be fatal in the context of a clinical application.

Purpose A novel finite-element model of ventricular torsion for the analysis of the twisting behavior of the left human ventricle was developed, in order to investigate the influence of various biomechanical parameters on cardiac kinematics.Methods The ventricle was simulated as a thick-walled ellipsoid composed of nine concentric layers. Arrays of reinforcement bars were embedded in each layer to mimic physiological myocardial anisotropy. The reinforcement bars were activated through an artificial combination of thermal and mechanical effects in order to obtain a contractile behavior which is similar to that of myocardial fibers. The presence of an incompressible fluid inside the ventricular cavity was also simulated and the ventricle was combined with simple lumped-parameter hydraulic circuits reproducing preload and afterload. Changes to a number of cardiac parameters, such as preload, afterload and fiber angle orientation were introduced, in order to study the effects of these changes on cardiac torsion.Results The model is able to reproduce a similar torsional behavior to that of a physiological heart. The results of the simulations showed that there was sound correspondence between the model outcomes and available data from the literature. Results confirmed the importance of symmetric transmural patterns for fiber orientation.Conclusions This model represents an important step on the path towards unveiling the complexity of cardiac torsion. It proves to be a practical and versatile tool which could assist clinicians and researchers by providing them with easily-accessible, detailed data on cardiac kinematics for future diagnostic and surgical purposes.

Background Sinonasal polyposis (SNP) is a chronic inflammatory pathology of nasal and paranasal cavities. Human leukocyte antigen (HLA) G molecules are nonclassic class I antigens with anti-inflammatory and tolerogenic properties. As most theories consider polyps to be the manifestation of chronic inflammation, there could be a possible implication of HLA-G molecules in SNP. The purpose of this study was to investigate the possible correlation between SNP and the production of soluble HLA-G (sHLA-G) by peripheral blood mononuclear cells (PBMCs).Methods The study involved 22 SNP patients (11 with no evidence of disease [NED] after surgery and 11 with relapse [RE]) and 20 healthy subjects. The presence of sHLA-G in PBMC lipopolysaccharide (LPS)-stimulated culture supernatants was analyzed. The levels of interleukin (IL) 10, one of the main up-regulators of sHLA-G production, were determined. Exogenous IL-10 was added to the SNP PBMC cultures to reconstitute the impairment in sHLA-G production.Results Increased IL-10 levels in LPS-activated PBMC culture supernatants were found in NED patients in comparison with healthy subjects (p = 0.0184). No sHLA-G production was observed in either of the patient subgroup supernatants (p < 0.0001). The addition of exogenous IL-10 showed the reconstitution of sHLA-G production in NED and in a lower amount in RE patients.Conclusion The results show a defect in sHLA-G production in SNP patients mainly related to the IL-10/HLA-G pathway. Given the anti-inflammatory functions of HLA-G molecules, this impairment could increase the susceptibility to the disease. The different sHLA-G production after exogenous IL-10 addition between NED and RE SNP could represent a marker of disease severity.

This paper presents a comparative study on the electrochemical behavior of the flavonoid rutin on a rigid carbon‐polyurethane composite electrode and on a glassy carbon electrode. The electrochemical oxidation reaction of rutin was found to be quasireversible and affected by adsorption on the electrode surface. A square‐wave voltammetric method was developed for determination of rutin in green tea infusion samples using the RCPE electrode and data treatment by a deconvolution procedure. The detection limit achieved in buffered solutions was 7.1×10−9mol L−1using the RCPE and 1.7×10−8mol L−1using the GC electrode the average reproducibility for five determinations being 3.5%.

The Atlantic Rainforest is a Brazilian ecosystem that is being rapidly being destroyed, along with the abiotic and biotic factors present in it. Among the biotic factors, the fungi are found in the soil which, besides being of major importance in terms of ecological niches, also have broad and significant applications in biotechnology. In order to assess the biodiversity of these microorganisms in this type of ecosystem, the Banhado Grande region was chosen at the Juréia-Itatins Ecology Station, in the state of São Paulo, Brazil. Within this region, two areas were delimited for study, one covered with natural (primary) vegetation and the other containing vegetation that regenerated following the planting of rice crops, referred to here as secondary. Collection of compound soil samples were taken (depth 0–15 cm) over a period of two and a half years, with the litter first being removed, during dry/cold and humid/hot periods. After sifting the samples, they were appropriately processed using the serial dilution technique to isolate the fungi from the soil. Six different culture media were used, having pHs of 4.5, 7.0 and 9.0. Altogether, 1,211 strains were isolated, divided into the following groups: Hyphomycetes, the most abundant followed by Ascomycetes, Zygomycetes, Coelomycetes, and Oomycetes. From these, 112 species were identified, 8 down to the genus level, and those that did not produce conidia were grouped as Mycelia sterilia. Among the strains, 67 were cellulolytic, 32 originated solely in soil under natural vegetation, and 26 originated solely in soil under secondary vegetation. (© 2005 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)

Background:Primary bare metal stenting and abciximab infusion are currently considered the best available reperfusion strategy for acute ST-segment elevation myocardial infarction (STEMI). Sirolimus eluting stents (SES), compared to bare metal stent (BMS), greatly reduce the incidence of binary restenosis and target vessel revascularisation (TVR), but their use on a routine basis results in a significant increase in medical costs. With current European list prices, the use of tirofiban instead of abciximab would save enough money to absorb the difference between SES and BMS. Aim:To assess whether in patients with STEMI the combination of SES with high dose bolus (HDB) tirofiban results in a similar incidence of major cardiovascular events (MACE) but in a lower binary restenosis rate after six months compared to BMS and abciximab. Methods and Results:160 patients are required to satisfy the primary composite end-point, including MACE and binary restenosis. The study is ongoing: the current paper focuses on the methodology and demography of the first 100 patients so far enrolled. Patients randomised to HDB tirofiban (n= 50, mean age: 62 ± 12, 40 males) and abciximab (n= 50, mean age: 63 ± 12, 38 males) do not differ for medical history, presentation profile, medications at discharge, angiographic profile and creatine-kinase MB-fraction at peak. Conclusions:The results of the trial will be available by the end of 2004: they will be crucial for the cardiologists to know whether the gold standard for AMI treatment should be reconsidered after the introduction of SES into the clinical practice.

AbstractThe effects of adding 2% of natural zeolite, containing a high percentage of clinoptilolite, to the diet of piglets was evaluated by monitoring growing performance and plasma parameters. The diet was also deprived of Colistine, an antibiotic usually added to piglet diets at subtherapeutic levels to prevent gastrointestinal pathology. Sixty-four piglets, weaned at 7.9 kg live weight, were divided in 2 groups of 32 each. The control group was given commercial piglet feedstuffs; for the treated group, 98% of the same feed, deprived of Colistine, was used, with the addition of 2% of a natural zeolite. The trial lasted 36 days. The piglets fed the control diet exhibited greater weight in the first three weeks, after which the differences were not significant. The average daily weight gain in the first week was higher in the control group (+37%; P<0.01) whereas at the end of the trial no differences were found. The feed intake did not show significant differences between groups and, as a result, the Feed Efficiency at the end of the trial was higher in the control group (532 vs. 491, P< 0.05). The plasmatic nitrogen parameters did not show significant differences between groups. Only in the first three weeks post-weaning the group fed the diet containing Colistine showed better ADG, subsequently it can be eliminated and clinoptilolite could favour growth.

In a search for retinoic acid (RA) receptor ligands endowed with potent apoptotic activity, a series of novel arotinoids were prepared. Because the stereochemistry of the C9-alkenyl portion of natural 9-cis-RA and the olefinic moiety of the previously synthesized isoxazole retinoid 4 seems to have particular importance for their apoptotic activity, novel retinoid analogues with a restricted or, vice versa, a larger flexibility in this region were designed and prepared. The new compounds were evaluated in vitro for their ability to activate natural retinoid receptors and for their differentiation-inducing activity. Cytotoxic and apoptotic activities were, in addition, evaluated. In general, these analogues showed low cytotoxicity, with the restricted structures being slightly more active than the more flexible ones. As an exception, however, the isoxazole retinoid 15b proved to be particularly able to induce apoptosis at concentrations &lt;5 μM, showing a higher activity than the classical retinoids such as all-trans-RA, 13-cis-RA, and 9-cis-RA and the previously described synthetic retinoid 4. 15b also exhibited a good affinity for the retinoid receptors. Interestingly, another important property of 15b was its ability to induce apoptosis in the HL60R multidrug-resistant (MDR) cell line, at the same concentration as is effective in HL60. Therefore, 15b represents a new retinoid possessing high apoptotic activity in an MDR cell line. The ability of 15b to act on K562 and HL60R cells suggests that this compound may have important implications in the treatment of different leukemias, and its structure could offer an interesting model for the design of new compounds endowed with apoptotic activity on MDR- and retinoid-resistant malignancies.

A convenient procedure to effect the Wittig and Horner−Wadsworth−Emmons reactions employs guanidine TBD and MTBD as base-promoters; mild reaction conditions, high efficiency, and facile isolation of the final products make the present methodology, at least in some cases, a practical alternative to known procedures.

The administration of cadmium, as food supplement, its bioaccumulation, and the effects on the development of tilapia Oreochromis niloticuswere investigated. The average size and weight and its behavior compared with controls were investigated during the period January 31, 1997, until March 31, 1999. At intervals of 60 days the measurements of size and weight were performed, and the concentration of cadmium in feces, water, muscular tissue, and viscera were determined by atomic absorption spectroscopy. The initial average cadmium concentration in food was 5 mg·kg−1and only after 6 months a small effect on size and weight could be observed. With increases in cadmium concentration to 50 mg·kg−1, beginning after the 7th month, and 100 mg·kg−1after the 16th month, a clear difference in size and weight and also in behavior could be observed. An LC50value of 40 mg·kg−1was observed after the 23rd month.

Nanoparticles are popular delivery vehicles, but their diffusional release results in inconstant drug delivery. Here, we flatten the delivery profile into a more constant, zero-order profile. Brain-derived neurotrophic factor (BDNF) is attached to photoactive titanium dioxide nanoparticles and loaded into a nanofibrous self-assembling peptide (SAP) hydrogel. Different UV exposure conditions show three distinct profiles, including a counterintuitive decrease in release after UV exposure. We propose that the adsorption of the freed growth factor onto the hydrogel nanofibers affects release. Nanoparticles diffuse from the hydrogel readily, carrying the bound growth factor, but the freed growth factor (released from the nanoparticles by UV) instead interacts with─and is released less readily from─the hydrogel. UV shifts growth factor from nanoparticles to the hydrogel, therefore changing the diffusional release. Through midpoint UV exposure, we achieve a flattened delivery profile─unusual for diffusion─by changing in situthe amount of growth factor bound to the diffusing nanoparticles. With nanoparticle diffusion alone, we observed an increasing release profile with 36% of release in the first 6 h and 64% in the second 6 h. With midway UV exposure, this was controlled to 49 and 51%, respectively. The release of an unbound (soluble) control growth factor, glial cell-line derived neurotrophic factor (GDNF), was not affected by UV treatment, demonstrating the potential for independent control of temporal delivery profiles in a multiagent material.

The expression of the ras and c-erbB2 oncoproteins (p21 and p185, respectively), together with estrogen receptor (ER) and progesterone receptor (PgR) determination, has been retrospectively analyzed in 68 primary breast carcinomas and in 19 normal breast tissue samples. The aims of this study were: a) to explore the association between ras and c-erbB2 expression; b) to evaluate the relationship between ras and c-erbB2 expression and both steroid receptor status and the classical clinical and pathological parameters; and c) to compare two different methods for p185 determination.p185 and p21 were measured by enzyme immunoassay (EIA); p185 was also determined by Western blotting (WB); ER and PgR were assayed by radioligand binding assay. The highest value of p185 in benign breast lesions was used as the threshold to distinguish between positive and negative samples. With this threshold the c-erbB2 oncoprotein was overexpressed in 41.2% (with EIA) and in 50% (with WB) of cancer samples. The concordance rate between the two methods was 79.4. No significant association was found between p21 and p185 levels either in cancer or in normal breast tissue samples. Increasing levels of tumor p21 were associated with a shorter time to recurrence and overall survival. Increasing levels of p185 were associated with a significantly shorter time to recurrence (p185 EIA: p=0.04, p185 WB: p=0.029) and overall survival (p185 EIA: p=0.04, p185 WB: p=0.029).