An integrative insight on dormancy alleviation in diaspores of Urochloa humidicola (Rendle) Morrone & Zuloaga, a tropical grass with great economic and ecological impact.

Even though exhaustively studied, dormancy alleviation in diaspores of Urochloa humidicola (Rendle) Morrone & Zuloaga remains a mystery. To clarify this, we asked the following question: could dormancy alleviation in diaspores of this species be associated with ageing, GA/ABA balance and remaining structures of the panicoid spikelet? We answer this question using diaspores of U. humidicola cv. BRS Tupi as a biological model, a cultivar with a wide edaphoclimatic range in Neotropical areas and whose diaspores possess 'deep dormancy' when dispersed. We analysed both germination and early plant development using a split-plot model. Our findings demonstrate that dormancy alleviation in diaspores of U. humidicola is a synergic phenomenon driven by crosstalk between age, GA/ABA balance and remaining structures of the panicoid spikelet covering caryopses, since this interaction acts on the dynamics of germination and early plant development. We demonstrate that: (i) spreading germination time is a maternal survival mechanism of this species, which has repercussions for occupational aggressiveness of the species; (ii) remaining structures of the panicoid spikelet covering caryopses are the main modulator of embryo development. These structures control the after-ripening process, which is modulated by some molecular factor. We also highlight that it is necessary to review concepts about dormancy of dispersal units in this grass species.

Antioxidant activity and chemical composition of oleoresin from leaves and flowers of Brunfelsia uniflora.

In this study, the temperature and pressure of supercritical CO2 extraction were evaluated to obtain oleoresin of Brunfelsia uniflora leaves and flowers. The oleoresin compounds were identified by gas chromatography-mass spectrometry. The antioxidant activity was evaluated by three different methods. The highest oleoresin yields were 3.32% at 40°C and 200 bar for the leaves, and 1.03% at 60°C and 200 bar for the flowers. The main extracted compounds from leaves were phytol varying from 11.95 to 36.42% and α-tocopherol from 15.53 to 43.10%, and from flowers were geranyl linalool from 11.05 to 21.42% and α-amyrin from 9.66 to 22.12%. Oleoresin obtained at 60°C and 150 bar from leaves presented high antioxidant activity by DPPH (IC50 1.90 mg/mL) and by FRAP (1.8 µmol Fe2+/mg). ß-carotene/linoleic acid co-oxidation oleoresin from leaves at 0.25 mg/mL presented higher antioxidant activity than Trolox. The total phenolic content of the oleoresin from leaves ranged from 66.20 to 83.33 µg/mg and from flowers it was just up to 12.46 µg/mg. The extraction conditions affected yield, chemical composition, and antioxidant activity of oleoresin from leaves and flowers. This is the first report on the antioxidant activity of B. uniflora oleoresin from leaves and flowers and provides subsidies for potential applications in chemical, pharmaceutical, and food industries.

Expression analysis of osteopontin mRNA splice variants in prostate cancer and benign prostatic hyperplasia.

Osteopontin splicing isoforms (OPN-SI) present differential expression patterns and specific tumor roles. Our aims were to characterize OPN-SI expression in prostate cancer (PCa) and benign prostate hyperplasia (BPH) tissues, besides evaluating their potential as biomarkers for PCa diagnosis and prognostic implications. Prostatic tissue specimens were obtained from 40 PCa and 30 benign prostate hyperplasia (BPH) patients. Quantitative real time PCR (qRT-PCR) was used to measure OPN-SI mRNA expression. Immunohistochemical analysis was performed using an anti-OPNc polyclonal antibody. Biostatistical analyses evaluated the association of OPN-SI and total Prostate Specific Antigen (PSA) serum levels with clinical and pathological data. PCa tissue samples presented significantly higher levels of OPNa, OPNb and OPNc transcripts (p<0.01) than in BPH specimens. OPN-SI mRNA expression were positively correlated with Gleason Score (p<0.01). ROC curves and logistic regression analyses demonstrated that OPN-SI and PSA were able to distinguish PCa from BPH patients (p<0.01). The OPNc isoform was the most upregulated variant and the best marker to distinguish patients' groups, presenting sensitivity and specificity of 90% and 100%, respectively. Immunohistochemistry analysis also demonstrated OPNc upregulation in PCa samples as compared to BPH tissues. OPNcprotein was also strongly stained PCa tissues presenting High Gleason Score. Multivariate analysis indicated that OPNc expression levels above the cut-off value presented a chance 4-fold higher for PCa occurrence. We conclude that OPN-SI were overexpressed in PCa tissues, strongly associated with PCa occurrence and with tumor cell differentiation. Our results suggest OPNc splicing isoform as an important biomarker contributing to improve PCa diagnosis and prognosis, besides providing insights into early steps of PCa carcinogenesis.

Equilibrium of Cu(II) and Ni(II) biosorption by marine alga Sargassum filipendula in a dynamic system: competitiveness and selectivity.

The study focuses on the equilibrium of dynamic biosorption in single and binary systems containing Cu(II) and Ni(II) ions using Sargassum filipendula (a marine alga). The experiments were performed in fixed-bed columns with both single-component and bi-component metal solutions (using different molar concentrations). Experimental data were fitted with different equilibrium models such as Langmuir, Langmuir with inhibition, Jain and Snowyink and Langmuir-Freundlich equations. The biosorption of pure metal ions in solution presented adequate capacities both for Cu(II) and Ni(II). In binary solutions the preferential sorption of Cu(II) over Ni(II) was demonstrated by the displacement of Ni(II) (marked overshoot on the breakthrough curves).

ABA inhibits embryo cell expansion and early cell division events during coffee (Coffea arabica 'Rubi') seed germination.

BACKGROUND AND AIMS: Coffee seed germination represents an interplay between the embryo and the surrounding endosperm. A sequence of events in both parts of the seed determines whether germination will be successful or not. Following previous studies, the aim here was to further characterize the morphology of endosperm degradation and embryo growth with respect to morphology and cell cycle, and the influence of abscisic acid on these processes. METHODS: Growth of cells in a fixed region of the axis was quantified from light micrographs. Cell cycle events were measured by flow cytometry and by immunocytochemistry, using antibodies against beta-tubulin. Aspects of the endosperm were visualized by light and scanning electron microscopy. KEY RESULTS: The embryonic axis cells grew initially by isodiametric expansion. This event coincided with reorientation and increase in abundance of microtubules and with accumulation of beta-tubulin. Radicle protrusion was characterized by a shift from isodiametric expansion to elongation of radicle cells and further accumulation of beta-tubulin. Early cell division events started prior to radicle protrusion. Abscisic acid decreased the abundance of microtubules and inhibited the growth of the embryo cells, the reorganization of the microtubules, DNA replication in the embryonic axis, the formation of a protuberance and the completion of germination. The endosperm cap cells had smaller and thinner cell walls than the rest of the endosperm. Cells in the endosperm cap displayed compression followed by loss of cell integrity and the appearance of a protuberance prior to radicle protrusion. CONCLUSIONS: Coffee seed germination is the result of isodiametric growth of the embryo followed by elongation, at the expense of integrity of endosperm cap cells. The cell cycle, including cell division, is initiated prior to radicle protrusion. ABA inhibits expansion of the embryo, and hence subsequent events, including germination.

Clinical outcome of long-term renal transplant survivors: a nephrourologic approach.

BACKGROUND: Long-term renal transplant survivors display high incidences of hypogonadism, neoplasms, erectile dysfunction, and metabolic syndrome. Although the urologist has a major role in the kidney transplantation surgery and treatment of the graft-related surgical complications, the follow-up of these patients is essentially focused on kidney function and the immunosuppressive regimen. We sought to evaluate the role of a nephrourologic follow-up for kidney transplant recipients. METHODS: We evaluated 27 male patients of mean age of 48.1 +/- 7.5 years and mean renal allograft follow-up of 12.3 +/- 4.3 years. The nephrourologic approach consisted of evaluation of the urinary system, prostate cancer screening, and assessment of voiding function, fertility, erectile dysfunction, hypogonadism, and metabolic syndrome. We also assessed quality of life. RESULTS: Major urologic findings were found in 18 (66.7%) patients. Four patients reported posttransplantation parenthood. The mean serum creatinine was 1.5 +/- 0.6 mg/dL and the calculated clearance, 72.6 +/- 27.4 mL/min/1.73 m(2). Regarding the voiding pattern, 92.6% of patients showed mild to moderate bladder outlet obstruction. Clinically significant increases in prostate volume were observed in 3.7% of patients. Mean serum total prostate-specific antigen was 1.6 +/- 1.5 ng/mL. Erectile dysfunction was displayed by 46.2% and hypogonadism by 74.1% of patients. Metabolic syndrome was diagnosed in 44.4% of patients, and quality of life was generally rated as good (4/5). CONCLUSION: Long-term renal transplantation survivors show high prevalences of treatable urologic diseases. Thus, we strongly recommend routine complete nephrourologic follow-up of this population.

Auto-antibodies in prostate cancer: humoral immune response to antigenic determinants coded by the differentially expressed transcripts FLJ23438 and VAMP3.

BACKGROUND: Here we evaluate auto-antibody response against two potential antigenic determinants of genes highly expressed in low Gleason Score prostate cancer (PC) tumor samples, namely FLJ23438 and VAMP3. METHODS: RT-PCR assays were used to analyze mRNA expression profiles of FLJ23438 and VAMP3 transcripts. The auto-antibody response against FLJ23438 and VAMP3 recombinant proteins was tested by immunoblot assays using PC, benign prostate hyperplasia (BPH), healthy donors (HD), and other human cancers plasma samples. RESULTS: Our data showed that 37% (10/27) and 7.4% (2/27) of PC plasma samples presented auto-antibodies against FLJ23438 and VAMP3, respectively. Only 8.3% (1/12) of BPH plasma samples were reactive for both auto-antibodies, while none (0/12) of HD plasma samples tested were reactive. CONCLUSIONS: The prevalence of 37% of positive PC plasma samples for anti-FLJ23438 antibodies suggests that humoral immune response against this antigenic determinant could be a potential serum marker for this cancer.

Exogenous gibberellins inhibit coffee (Coffea arabica cv. Rubi) seed germination and cause cell death in the embryo.

The mechanism of inhibition of coffee (Coffea arabica cv. Rubi) seed germination by exogenous gibberellins (GAs) and the requirement of germination for endogenous GA were studied. Exogenous GA(4+7) inhibited coffee seed germination. The response to GA(4+7) showed two sensitivity thresholds: a lower one between 0 and 1 microM and a higher one between 10 and 100 microM. However, radicle protrusion in coffee seed depended on the de novo synthesis of GAs. Endogenous GAs were required for embryo cell elongation and endosperm cap weakening. Incubation of coffee seed in exogenous GA(4+7) led to loss of embryo viability and dead cells were observed by low temperature scanning microscopy only when the endosperm was surrounding the embryo. The results described here indicate that the inhibition of germination by exogenous GAs is caused by factors that are released from the endosperm during or after its weakening, causing cell death in the embryo and leading to inhibition of radicle protrusion.

Abscisic acid controls embryo growth potential and endosperm cap weakening during coffee (Coffea arabica cv. Rubi) seed germination.

The mechanism and regulation of coffee seed germination were studied in Coffea arabica L. cv. Rubi. The coffee embryo grew inside the endosperm prior to radicle protrusion and abscisic acid (ABA) inhibited the increase in its pressure potential. There were two steps of endosperm cap weakening. An increase in cellulase activity coincided with the first step and an increase in endo-beta-mannanase (EBM) activity with the second step. ABA inhibited the second step of endosperm cap weakening, presumably by inhibiting the activities of at least two EBM isoforms and/or, indirectly, by inhibiting the pressure force of the radicle. The increase in the activities of EBM and cellulase coincided with the decrease in the force required to puncture the endosperm and with the appearance of porosity in the cell walls as observed by low-temperature scanning electronic microscopy. Tissue printing showed that EBM activity was spatially regulated in the endosperm. Activity was initiated in the endosperm cap whereas later during germination it could also be detected in the remainder of the endosperm. Tissue printing revealed that ABA inhibited most of the EBM activity in the endosperm cap, but not in the remainder of the endosperm. ABA did not inhibit cellulase activity. There was a transient rise in ABA content in the embryo during imbibition, which was likely to be responsible for slow germination, suggesting that endogenous ABA also may control embryo growth potential and the second step of endosperm cap weakening during coffee seed germination.

Demonstration of apoptosis in human skin injuries as an indicator of vital reaction.

Demonstrating the vital character of an injury and estimation of the age are routine tasks in forensic pathology and although many different techniques have been applied to this problem none have been found to be completely satisfactory. Apoptosis, an active genetically controlled process, is the major mechanism by which homeostasis of a number of physiological systems in the body is regulated and changes in the rate following different kinds of stimuli have prompted us to test it as an indicator of vitality. We used an in situ end-labelling technique (Apop-Tag) in 30 human surgical skin injuries with age since injury ranging from 3 min to 8 h and found that apoptotic keratinocytes are found in over 50% of the cases with a post-infliction interval of at least 120 min. Apoptosis was not seen in injuries less than 120 min old or in normal skin, which was used as an external control. These results suggest that apoptosis could be a useful indicator for the intravital occurrence of injuries and could help to estimate the date of the skin injuries in some cases. The importance of strict technical control is stressed and the necessity of a complementary technique to confirm apoptosis is discussed.

Immunohistochemical detection of fibronectin and tenascin in incised human skin injuries.

Immunohistochemical detection of molecules involved in inflammatory reaction can be useful for the diagnosis of vitality in skin wounds. We studied the expression of fibronectin (FN) and tenascin (TN) in 58 human skin wounds (48 vital and 10 postmortem). The age of vital injuries ranged from 3 min to 8 h and postmortem specimens were collected after a postinfliction interval of 15-180 min. One hundred thirty-seven formalin-fixed paraffin-embedded sections (mean: 2.3 sections per case) were stained with each of two monoclonal antibodies against FN and TN using the streptABC technique. A reticular staining for FN in wound edge and dermis was observed in 50% of vital specimens versus 0% in postmortem cases. Immunoreactivity was reduced in 10 autolysed cases. FN positivity exclusively at the injury margin was observed in 39.4% of vital wounds and 10% of postmortem cases. TN was negative in all specimens. Vital and postmortem hemorrhage areas showed positivity for FN and TN. Due to its low sensitivity, immunohistochemical analysis of FN is useful for determining vitality only in a minority of cases. Different factors in everyday practice, including autolysis and technical problems often produce false negative reactions with the result that FN cannot be regarded as a reliable parameter of vitality. Positive reactions (network staining) are more valuable than negativity but are not pathognomonic. Both vital and postmortem hemorrhages show an enhanced positivity for FN and TN, thus impeding the diagnosis.

[Urethral substitution with synthetic material]. / Sustitución de la uretra con material sintético.

In spite of the numerous surgical techniques described, management of urethral stenosis continues to be an unresolved problem. Free graft urethroplasty is indicated in selected cases and several organic and synthetic materials have been described for this use. Our group reviews the synthetic alloplasts used for partial or total replacement of the male urethra. The search for an appropriate alloplast for urethral replacement has been rather frustrating. Complication rates are still too high to allow routine usage. The most encouraging results are those with new absorbable materials which cause minimal inflammatory reactions of a foreign body type. These are readily available and allow urethral replacement using tissues regenerated from removed ends.

Intracranial EEG versus flumazenil and glucose PET in children with extratemporal lobe epilepsy.

OBJECTIVE: To compare abnormalities determined in 2-deoxy-2-[18F]fluoro-D-glucose (FDG) and [11C]flumazenil (FMZ) PET images with intracranial EEG data in patients with extratemporal lobe epilepsy. BACKGROUND: Although PET studies with FDG and FMZ are being used clinically to localize epileptogenic regions in patients with refractory epilepsy, the electrophysiologic significance of the identified PET abnormalities remains poorly understood. METHODS: We studied 10 patients, mostly children (4 boys, 6 girls, aged 2 to 19 years; mean age, 11 years), who underwent FDG and FMZ PET scans, intracranial EEG monitoring, and cortical resection for intractable epilepsy. EEG electrode positions relative to the brain surface were determined from MRI image volumes. Cortical areas of abnormal glucose metabolism or FMZ binding were determined objectively based on asymmetry measures derived from homotopic cortical areas at three asymmetry thresholds. PET data were then coregistered with the MRI and overlaid on the MRI surface. A receiver operating characteristics (ROC) analysis was performed to determine the specificity and sensitivity of PET-defined abnormalities against the gold standard of intracranial EEG data. RESULTS: FMZ PET detected at least part of the seizure onset zone in all subjects, whereas FDG PET failed to detect the seizure onset region in two of 10 patients. The area under the ROC curves was higher for FMZ than FDG PET for both seizure onset (p = 0.01) and frequent interictal spiking (p = 0.04). Both FMZ and FDG PET showed poor performance for detection of rapid seizure spread (area under the ROC curve not significantly different from 0.5). CONCLUSIONS: [11C]flumazenil (FMZ) PET is significantly more sensitive than 2-deoxy-2-[18F]fluoro-D-glucose (FDG) PET for the detection of cortical regions of seizure onset and frequent spiking in patients with extratemporal lobe epilepsy, whereas both FDG and FMZ PET show low sensitivity in the detection of cortical areas of rapid seizure spread. The application of PET, in particular FMZ PET, in guiding subdural electrode placement in refractory extratemporal lobe epilepsy will enhance coverage of the epileptogenic zone.

Concepts of dependence among Brazilian alcohol- and drug-dependent patients.

Studies suggest that the concepts that drug dependent persons have about their problems could influence adherence to treatment and outcome. The objective of this study was to compare these concepts in different groups of patients. Three groups of drug-dependent patients (diagnosed according to DSM-III-R criteria) were compared: 64 dependent on alcohol only, 56 dependent only on other drugs and 39 dependent on both alcohol and drugs. They were asked: "What do you think about your problem?" and the responses were qualitatively analyzed and classified into themes. For the whole sample, the main themes mentioned were: the function of alcohol and drugs in their lives (50%), the etiology of dependence (32%) and reasons for initial use (21%). Patients dependent on alcohol associated their use with coping, while patients dependent on other drugs reported pleasure seeking. In regard to etiology, most patients classified their dependence as a habit. Patients dependent on alcohol tended to attribute an organic etiology to their problem, while those dependent on other drugs most frequently considered moral aspects as important. This study suggests that there are differences between these three groups in the appraisal of their problem. Professionals interested in treatment matching need to take these findings into account, as they may influence compliance and treatment outcome.