Thymocyte migration: an affair of multiple cellular interactions?

Cell migration is a crucial event in the general process of thymocyte differentiation. The cellular interactions involved in the control of this migration are beginning to be defined. At least chemokines and extracellular matrix proteins appear to be part of the game. Cells of the thymic microenvironment produce these two groups of molecules, whereas developing thymocytes express the corresponding receptors. Moreover, although chemokines and extracellular matrix can drive thymocyte migration per se, a combined role for these molecules appears to contribute to the resulting migration patterns of thymocytes in their various stages of differentiation. The dynamics of chemokine and extracellular matrix production and degradation is not yet well understood. However, matrix metalloproteinases are likely to play a role in the breakdown of intrathymic extracellular matrix contents. Thus, the physiological migration of thymocytes should be envisioned as a resulting vector of multiple, simultaneous and/or sequential stimuli involving chemokines, adhesive and de-adhesive extracellular matrix proteins, as well as matrix metalloproteinases. Accordingly, it is conceivable that any pathological change in any of these loops may result in the alteration of normal thymocyte migration. This seems to be the case in murine infection by the protozoan parasite Trypanosoma cruzi, the causative agent of Chagas' disease. A better knowledge of the physiological mechanisms governing thymocyte migration will provide new clues for designing therapeutic strategies targeting developing T cells.

A method for multiple sequential analyses of macrophage functions using a small single cell sample

Microbial pathogens such as bacillus Calmette-Guérin (BCG) induce the activation of macrophages. Activated macrophages can be characterized by the increased production of reactive oxygen and nitrogen metabolites, generated via NADPH oxidase and inducible nitric oxide synthase, respectively, and by the increased expression of major histocompatibility complex class II molecules (MHC II). Multiple microassays have been developed to measure these parameters. Usually each assay requires 2-5 x 10(5) cells per well. In some experimental conditions the number of cells is the limiting factor for the phenotypic characterization of macrophages. Here we describe a method whereby this limitation can be circumvented. Using a single 96-well microassay and a very small number of peritoneal cells obtained from C3H/HePas mice, containing as little as <=2 x 10(5) macrophages per well, we determined sequentially the oxidative burst (H2O2), nitric oxide production and MHC II (IAk) expression of BCG-activated macrophages. More specifically, with 100 µl of cell suspension it was possible to quantify H2O2 release and nitric oxide production after 1 and 48 h, respectively, and IAk expression after 48 h of cell culture. In addition, this microassay is easy to perform, highly reproducible and more economical.

Gene expression profiling of clinical stages II and III breast cancer

Clinical stage (CS) is an established indicator of breast cancer outcome. In the present study, a cDNA microarray platform containing 692 genes was used to identify molecular differences between CSII and CSIII disease. Tumor samples were collected from patients with CSII or CSIII breast cancer, and normal breast tissue was collected from women without invasive cancer. Seventy-eight genes were deregulated in CSIII tumors and 22 in CSII tumors when compared to normal tissue, and 20 of them were differentially expressed in both CSII and CSIII tumors. In addition, 58 genes were specifically altered in CSIII and expression of 6 of them was tested by real time RT-PCR in another cohort of patients with CSII or CSIII breast cancer and in women without cancer. Among these genes, MAX, KRT15 and S100A14, but not APOBEC3G or KRT19, were differentially expressed on both CSIII and CSII tumors as compared to normal tissue. Increased HMOX1 levels were detected only in CSIII tumors and may represent a molecular marker of this stage. A clear difference in gene expression pattern occurs at the normal-to-cancer transition; however, most of the differentially expressed genes are deregulated in tumors of both CS (II and III) compared to normal breast tissue.

Application of a modified sequential organ failure assessment score to critically ill patients

The purpose of the present study was to explore the usefulness of the Mexican sequential organ failure assessment (MEXSOFA) score for assessing the risk of mortality for critically ill patients in the ICU. A total of 232 consecutive patients admitted to an ICU were included in the study. The MEXSOFA was calculated using the original SOFA scoring system with two modifications: the PaO2/FiO2 ratio was replaced with the SpO2/FiO2 ratio, and the evaluation of neurologic dysfunction was excluded. The ICU mortality rate was 20.2%. Patients with an initial MEXSOFA score of 9 points or less calculated during the first 24 h after admission to the ICU had a mortality rate of 14.8%, while those with an initial MEXSOFA score of 10 points or more had a mortality rate of 40%. The MEXSOFA score at 48 h was also associated with mortality: patients with a score of 9 points or less had a mortality rate of 14.1%, while those with a score of 10 points or more had a mortality rate of 50%. In a multivariate analysis, only the MEXSOFA score at 48 h was an independent predictor for in-ICU death with an OR = 1.35 (95%CI = 1.14-1.59, P < 0.001). The SOFA and MEXSOFA scores calculated 24 h after admission to the ICU demonstrated a good level of discrimination for predicting the in-ICU mortality risk in critically ill patients. The MEXSOFA score at 48 h was an independent predictor of death; with each 1-point increase, the odds of death increased by 35%.

Elevated levels of plasma osteoprotegerin are associated with all-cause mortality risk and atherosclerosis in patients with stages 3 to 5 chronic kidney disease

Osteoprotegerin (OPG) regulates bone mass by inhibiting osteoclast differentiation and activation, and plays a role in vascular calcification. We evaluated the relationship between osteoprotegerin levels and inflammatory markers, atherosclerosis, and mortality in patients with stages 3-5 chronic kidney disease. A total of 145 subjects (median age 61 years, 61% men; 36 patients on hemodialysis, 55 patients on peritoneal dialysis, and 54 patients with stages 3-5 chronic kidney disease) were studied. Clinical characteristics, markers of mineral metabolism (including fibroblast growth factor-23 [FGF-23]) and inflammation (high-sensitivity C-reactive protein [hsCRP] and interleukin-6 [IL-6]), and the intima-media thickness (IMT) in the common carotid arteries were measured at baseline. Cardiac function was assessed by color tissue Doppler echocardiography. After 36 months follow-up, the survival rate by Kaplan-Meier analysis was significantly different according to OPG levels (χ2=14.33; P=0.002). Increased OPG levels were positively associated with IL-6 (r=0.38, P<0.001), FGF-23 (r=0.26, P<0.001) and hsCRP (r=0.0.24, P=0.003). In addition, OPG was positively associated with troponin I (r=0.54, P<0.001) and IMT (r=0.39, P<0.0001). Finally, in Cox analysis, only OPG (HR=1.07, 95%CI=1.02-1.13) and hsCRP (HR=1.02, 95%CI=1.01-1.04) were independently associated with increased risk of death. These results suggested that elevated levels of serum OPG might be associated with atherosclerosis and all-cause mortality in patients with chronic kidney disease.

More than 10 years survival with sequential therapy in a patient with advanced renal cell carcinoma: a case report

Although radical nephrectomy alone is widely accepted as the standard of care in localized treatment for renal cell carcinoma (RCC), it is not sufficient for the treatment of metastatic RCC (mRCC), which invariably leads to an unfavorable outcome despite the use of multiple therapies. Currently, sequential targeted agents are recommended for the management of mRCC, but the optimal drug sequence is still debated. This case was a 57-year-old man with clear-cell mRCC who received multiple therapies following his first operation in 2003 and has survived for over 10 years with a satisfactory quality of life. The treatments given included several surgeries, immunotherapy, and sequentially administered sorafenib, sunitinib, and everolimus regimens. In the course of mRCC treatment, well-planned surgeries, effective sequential targeted therapies and close follow-up are all of great importance for optimal management and a satisfactory outcome.

Challenges of user-driven innovations on late stages of innovation process: evidence from ICT companies

The goal of this thesis is to study user-driven innovations and user involvement throughout the innovation process in context of B2B companies. Significant emphasis in the analysis put onto the late stages of innovation process and commercialization of innovations. Thesis includes detailed review of theoretical concepts and underlying frameworks of innovation process, lead users and user-driven innovations. The empirical part of the thesis consist of interviews of the four companies from ICT industry, followed by the comprehensive analysis and comparison of the results. The presented findings indicate common challenges, which ICT companies face, when shifting towards innovation by users paradigm. Linkages and connections among current situation and theoretical frameworks presented in the discussion part of the thesis allow to draw practical managerial implications. The results of the research emphasize valuable insights and challenges of user interactions within innovation process as well as output and participation related benefits for the companies and users. The research points out current state of the user involvement techniques and tools used for user interactions as well as suggests the possibilities for improvement in the future.

Volatile compounds in noni (Morinda citrifolia L.) at two ripening stages

The volatile components of noni at two ripening stages were isolated by headspace solid-phase microextraction using 65 µm Polydimethylsiloxane-Divinylbenzene (PDMS/DVB) fibers and analyzed using GC/MS. Both maturation stages had several compounds in common. Ninety-six compounds were identified, from which octanoic acid ( 70% of total extract) and hexanoic acid ( 8% of total extract) were found to be the major constituents. Due to noni maturation, octanoic acid, decanoic acid and 2E-nonenal decreased their concentrations, while some esters (methyl hexanoate, methyl octanoate, ethyl octanoate and methyl 4E-decenoate), which their fruity odor notes, increased their contents. Two unsaturated esters, reported for the first time in this fruit, 3-methyl-3-buten-1-yl hexanoate and 3-methyl-3-buten-1-yl octanoate, significantly decreased their concentration in the ripe to over-ripe fruits.

Chemical characterization of leaves of organically grown carrot Dacus carota L.) in various stages of development for use as food

In Brazil, street markets and vegetable distributors discard vegetable leaves and stems, including those of carrot (Dacus carota L.). Seeking to reduce the waste of vegetable parts, this study characterized chemically the leaves of organically grown carrot in three stages of development to determine the best time for their removal and consumption as food. The leaves were dehydrated in an oven at 70 °C for 43 hours and analyzed for chemical composition, antioxidant activity, chlorophyll content, fatty acid composition, and also calcium (Ca), sodium (Na), potassium (K), magnesium (Mg), manganese (Mn), iron (Fe), zinc (Zn), and copper (Cu) contents. The analyses indicated 100 days of development as the ideal stage for the removal and consumption of carrot leaves with good antioxidant activity requiring only 63.78 ± 0.5 mg.L-1 methanol leaf extract to inhibit 50% of the concentration of the free radical DPPH (2,2-diphenyl-1picrilidrazil), and total protein and alpha-linolenic acid (18:3 n-3/LNA) contents of 18.23% ± 2.8 and 876.55 ± 20.62 mg.100 g-1 of dry matter, respectively.

Changes in enzymes, phenolic compounds, tannins, and vitamin C in various stages of jambolan (Syzygium cumini Lamark) development

The physiological state of a fruit is closely related to ripening and climatic conditions during the growing period when the fruit undergo changes in color, texture, and flavor. The ripening of the fruit can involve a complex series of biochemical reactions with alteration in enzymes activities, phenols, tannins, and ascorbic acid. The activity of enzymes (carboximethylcellulase, polygalacturonase, and pectinlyase), the total concentration of phenolic compounds, condensed tannins, and vitamin C in five stages of maturation were studied. Significant changes were observed between the maturity stages. The phenolic compounds were higher at green stage (705.01 ± 7.41); tannins were higher at green/purple stage (699.45 ± 0.22). The results showed that the ascorbic acid levels of the pulp varied significantly from 50.81 ± 1.43 to 6.61 ± 1.04 mg.100 g-1 during maturation. The specific activity of pectin lyase was higher at green stage (1531.90 ± 5.83). The specific activity of polygalacturonase was higher at mature stage (1.83 ± 0.0018). The specific activity of carboximetilcelulose was higher at ripe mature stage (4.61 ± 0.0024). The low ascorbic acid content found in jambolan fruit indicates that this fruit is not a rich source of this nutrient; however, other characteristics can make jambolan products fit for human consumption.

Physicochemical changes in cubiu fruits (Solanum sessiliflorum Dunal) at different ripening stages

Cubiu shrubs (Solanum sessiliflorum Dunal) have drawn the attention of researchers for their biological versatility (preferential heliophilous or facultative ombrophilous shrubs), their capacity to grow in upland or lowland areas, and the good technological quality of their fruits for the food industry. The aim of this study was to verify physicochemical changes in cubiu fruits during maturation. The fruits were harvested from the experimental station of olericulture of the Instituto Nacional de Pesquisas da Amazônia (INPA), Brazil. The analyses were performed in whole cubiu fruits (peel, pulp, and placenta) at four traditional ripening stages (green, turning, ripe, and fully ripe) for the determination of weight, moisture, total solids, total carotenoids, proteins, lipids, and ash. Cubiu fruits showed large weight variation, with amodal distribution. The ripe stage was critical to maintain moisture, and from that stage on, water loss became evident. The lipids increased steadily over the four ripening stages, maintaining, however, insignificant calorie content. Total carotenoids, proteins, and ash reached the maximum level at the fully ripe stage. With the exception of weight and moisture, all physicochemical changes exhibited the same general behavior, i.e. they increased as the fruits ripened at the four investigated stages.

Prevalent fatty acids in cashew nuts obtained from conventional and organic cultivation in different stages of processing

Brazil is one of the three largest producers of fruits in the world, and among those fruit trees, the cashew tree stands out due to the high nutritional and commercial value of its products. During its fruit processing, there are losses in some compounds and few studies address this issue. Over the last decade the conventional system of food production has been substituted for the organic cultivation system, which is a promising alternative source of income given the global demand for healthy food. Therefore, this research aimed to characterize and quantify the prevalent fatty acids found in cashew nuts obtained from conventional and organic cultivation during various stages of processing. The prevalent fatty acids found were palmitic, linoleic, oleic, and stearic acid. The average of these fatty acids were 6.93 ± 0.55; 16.99 ± 0.61; 67.62 ± 1.00 and 8.42 ± 0.55 g/100 g, respectively. There was no reduction in the palmitic, oleic and stearic fatty acid contents during processing. Very little difference was observed between the nuts obtained from conventional and organic cultivation, indicating that the method of cultivation used has little or no influence on the content of cashew nut fatty acids.

Chemical and physical characterization of mume fruit collected from different locations and at different maturity stages in São Paulo State

Prunus mume is widely studied due to its health benefits regarding increase of blood fluidity and consequent improvement of the cardiovascular system and the prevention or even the fight against different types of cancer. However, in Brazil this culture is found only among oriental descendants. The present study aimed to characterize mume fruit collected from three different locations in the State of São Paulo regarding general aspects such as pH, total titratable acidity (TTA), total soluble solids (TTS), pectin content and yield of pulp and chemical characteristics: total phenolic compounds (TPC) and antioxidant capacity. Mume fruit were collected unripe and analyzed until maturation about 88 days after flowering. Fruit collected in Botucatu came from a commercial mume fruit producer and had average weight of 16.9 g, while in fruit from other locations weight varied from 5.7-6.9 g. TSS ranged from 9.5 to 10.0 Brix, total solids was 10.2-12.2% and pH showed values between 2.5 and 2.7 for all locations. TTA expressed in citric acid decreased from 4.0-5.7 g (100g- 1) at unripe stage to 2.0-3.8 g (100g- 1) in mature-stage fruit. Pectin content decreased from 11.2 to 10.8% during fruit maturation, TPC content was 147-226 mg catechin (g- 1) on a dry matter basis and the antioxidant capacity was 96-169 µMol Trolox (g- 1) on a dry matter basis or 21-34 µMol Trolox (g- 1) on a wet matter basis.

Evaluation of beetroot (Beta vulgaris L.) leaves during its developmental stages: a chemical composition study

Beetroot leaves (Beta vulgaris L.) are commonly cut off and discarded before using its bulb due to lack of knowledge of how to use them. Aiming at using these leaves, in the present study, in natura and dehydrated beetroot leaves were chemically characterized in terms of fatty acid composition, proximate composition, minerals, total phenolic compounds (TPC), and antioxidant activity by DPPH• in different stages (60, 80, and 100 days) of development. The beetroot leaves showed significant levels of protein and lipids in all developmental stages, and all proximate composition nutrients decreased during these maturation stages; the highest content was observed at 60 days. The Fe content decreased during the developmental stages (from 342.75 to 246.30 mg.kg-1), while the content of K increased (from 13,367.64 to 20,784.90 mg.kg-1). With regard to to fatty acid composition, linolenic acid was present in the greatest quantity, and it increase up to 2.58 mg.g-1 (in natura) and 40.11 mg.g-1 (dehydrated) at 100 days of development. The n-6/n-3 ratios were low in all stages. The TPC and antioxidant activity by DPPH• changed during the developmental stages. The TPC was highest in the 100-day dehydrated leaves (15.27±0.12 mg GAE.g-1 FW), and the 50% inhibition of DPPH• (IC50 89.52 µg.mL-1) were better in the 60-day in natura leaves. This study shows that all developmental stages produced satisfactory results, and therefore, these leaves can be reused as food. The antioxidant activity and the chemical constituents, mainly the ω-3fatty acid, increased during the stages of development.

Effects of sequential enzymatic hydrolysis on structural, bioactive and functional properties of Phaseolus lunatus protein isolate

Significant initiatives exist within the global food market to search for new, alternative protein sources with better technological, functional, and nutritional properties. Lima bean (Phaseolus lunatus L.) protein isolate was hydrolyzed using a sequential pepsin-pancreatin enzymatic system. Hydrolysis was performed to produce limited (LH) and extensive hydrolysate (EH), each with different degrees of hydrolysis (DH). The effects of hydrolysis were evaluated in vitro in both hydrolysates based on structural, functional and bioactive properties. Structural properties analyzed by electrophoretic profile indicated that LH showed residual structures very similar to protein isolate (PI), although composed of mixtures of polypeptides that increased hydrophobic surface and denaturation temperature. Functionality of LH was associated with amino acid composition and hydrophobic/hydrophilic balance, which increased solubility at values close to the isoelectric point. Foaming and emulsifying activity index values were also higher than those of PI. EH showed a structure composed of mixtures of polypeptides and peptides of low molecular weight, whose intrinsic hydrophobicity and amino acid profile values were associated with antioxidant capacity, as well as inhibiting angiotensin-converting enzyme. The results obtained indicated the potential of Phaseolus lunatus hydrolysates to be incorporated into foods to improve techno-functional properties and impart bioactive properties.