Evidence for the endophytic colonization of Phaseolus vulgaris(common bean) roots by the diazotroph Herbaspirillum seropedicae

Herbaspirillum seropedicae is an endophytic diazotrophic bacterium, which associates with important agricultural plants. In the present study, we have investigated the attachment to and internal colonization of Phaseolus vulgaris roots by the H. seropedicae wild-type strain SMR1 and by a strain of H. seropedicae expressing a red fluorescent protein (DsRed) to track the bacterium in the plant tissues. Two-day-old P. vulgaris roots were incubated at 30°C for 15 min with 6 x 10(8) CFU/mL H. seropedicae SMR1 or RAM4. Three days after inoculation, 4 x 10(4) cells of endophytic H. seropedicae SMR1 were recovered per gram of fresh root, and 9 days after inoculation the number of endophytes increased to 4 x 10(6) CFU/g. The identity of the recovered bacteria was confirmed by amplification and sequencing of the 16SrRNA gene. Furthermore, confocal microscopy of P. vulgaris roots inoculated with H. seropedicae RAM4 showed that the bacterial cells were attached to the root surface 15 min after inoculation; fluorescent bacteria were visible in the internal tissues after 24 h and were found in the central cylinder after 72 h, showing that H. seropedicae RAM4 is capable of colonizing the roots of the dicotyledon P. vulgaris. Determination of dry weight of common bean inoculated with H. seropedicae SMR1 suggested that this bacterium has a negative effect on the growth of P. vulgaris.

An MD-PhD program in Brazil: students’ concepts of science and of common sense

In 1995, a pioneering MD-PhD program was initiated in Brazil for the training of medical scientists in experimental sciences at the Federal University of Rio de Janeiro. The program’s aim was achieved with respect to publication of theses in the form of papers with international visibility and also in terms of fostering the scientific careers of the graduates. The expansion of this type of program is one of the strategies for improving the preparation of biomedical researchers in Brazil. A noteworthy absence of interest in carrying out clinical research limits the ability of young Brazilian physicians to solve biomedical problems. To understand the students’ views of science, we used qualitative and quantitative triangulation methods, as well as participant observation to evaluate the students’ concepts of science and common sense. Subjective aspects were clearly less evident in their concepts of science. There was a strong concern about "methodology", "truth" and "usefulness". "Intuition", "creativity" and "curiosity" were the least mentioned thematic categories. Students recognized the value of intuition when it appeared as an explicit option but they did not refer to it spontaneously. Common sense was associated with "consensus", "opinion" and ideas that "require scientific validation". Such observations indicate that MD-PhD students share with their senior academic colleagues the same reluctance to consider common sense as a valid adjunct for the solution of scientific problems. Overcoming this difficulty may be an important step toward stimulating the interest of physicians in pursuing experimental research.

Expression of β-catenin and c-myc during human common bile duct development: a possible role in the morphogenesis of the common bile duct

β-catenin and c-myc play important roles in the development of tissues and organs. However, little is known about their expression patterns during the development of the human common bile duct. Immunohistochemistry was used to detect β-catenin and c-myc expression in common bile duct samples from postmortem tissues of 14 premature infants and 6 spontaneously aborted fetuses. The expression of β-catenin and c-myc was also analyzed by Western blot. The samples were divided into four groups based on the stage of human fetal development: 12, 13-27, 28-37, and >37 weeks. The Image-Pro Plus v. 6.0 image analysis software was used to calculate the mean qualifying score (MQS). At fetal stages 12, 13-27, 28-37, and >37 weeks, MQS of β-catenin were 612.52±262.13, 818.38±311.73, 706.33±157.19, and 350.69±110.19, respectively. There was a significant difference in MQS among the four groups (ANOVA, P=0.0155) and between the scores at >37 and 13-27 weeks (Student-Newman-Keuls, P<0.05). At fetal stages 12, 13-27, 28-37, and >37 weeks, the MQS of c-myc were 1376.64±330.04, 1224.18±171.66, 1270.24±320.75, and 741.04±219.19, respectively. There was a significant difference in MQS among the four groups (ANOVA, P=0.0087) and between the scores at >37 and 12 weeks, >37 and 13-27 weeks, and >37 and 28-37 weeks (all P<0.05, Student-Newman-Keuls). Western blots showed that β-catenin and c-myc expression were significantly higher in fetal than in postnatal control duct tissue (P<0.05). c-myc and β-catenin are involved in the normal development of the human common bile duct.

Voluntary exercise prior to traumatic brain injury alters miRNA expression in the injured mouse cerebral cortex

MicroRNAs (miRNAs) may be important mediators of the profound molecular and cellular changes that occur after traumatic brain injury (TBI). However, the changes and possible roles of miRNAs induced by voluntary exercise prior to TBI are still not known. In this report, the microarray method was used to demonstrate alterations in miRNA expression levels in the cerebral cortex of TBI mice that were pretrained on a running wheel (RW). Voluntary RW exercise prior to TBI: i) significantly decreased the mortality rate and improved the recovery of the righting reflex in TBI mice, and ii) differentially changed the levels of several miRNAs, upregulating some and downregulating others. Furthermore, we revealed global upregulation of miR-21, miR-92a, and miR-874 and downregulation of miR-138, let-7c, and miR-124 expression among the sham-non-runner, TBI-non-runner, and TBI-runner groups. Quantitative reverse transcription polymerase chain reaction data (RT-qPCR) indicated good consistency with the microarray results. Our microarray-based analysis of miRNA expression in mice cerebral cortex after TBI revealed that some miRNAs such as miR-21, miR-92a, miR-874, miR-138, let-7c, and miR-124 could be involved in the prevention and protection afforded by voluntary exercise in a TBI model.

Is a contrast study really necessary prior to ureteroscopy?

This study aimed to evaluate the effect of preoperative imaging techniques on the success and complication rates of ureteroscopy. We performed a retrospective analysis of 736 patients (455 males and 281 females), with a mean age of 45.5±15.2 years (range, 1-88 years), who underwent rigid ureteroscopic procedures for removal of ureteral stones. Patients were divided into 4 groups according to the type of imaging modality used: group I, intravenous urography (n=116); group II, computed tomography (n=381); group III, computed tomography and intravenous urography (n=91), and group IV, ultrasonography and abdominal plain film (n=148). Patients’ demographics, stone size and location, prior shock wave lithotripsy, lithotripsy technique, operation time, success rate, and rate of intraoperative complications were compared among the groups. There were no significant differences in success and complication rates among the groups. The stone-free rate after primary ureteroscopy was 87.1% in group I, 88.2% in group II, 96.7% in group III, and 89.9% in group IV (P=0.093). The overall incidence of intraoperative complications was 11.8%. According to the modified Satava classification system, 6.1% of patients had grade 1, 5.1% had grade 2, and 0.54% had grade 3 complications. Intraoperative complications developed in 12.1% of patients in group I, 12.6% of patients in group II, 7.7% of patients in group III, and 12.2% of patients in group IV (P=0.625). Our findings clearly demonstrate that ureteroscopic treatment of ureteral stones can be safely and effectively performed with no use of contrast study imaging, except in doubtful cases of anatomical abnormalities.

High-impact exercise in rats prior to and during suspension can prevent bone loss

High-impact exercise has been considered an important method for treating bone loss in osteopenic experimental models. In this study, we investigated the effects of osteopenia caused by inactivity in femora and tibiae of rats subjected to jump training using the rat tail suspension model. Eight-week-old female Wistar rats were divided into five groups (n=10 each group): jump training for 2 weeks before suspension and training during 3 weeks of suspension; jump training for 2 weeks before suspension; jump training only during suspension; suspension without any training; and a control group. The exercise protocol consisted of 20 jumps/day, 5 days/week, with a jump height of 40 cm. The bone mineral density of the femora and tibiae was measured by double energy X-ray absorptiometry and the same bones were evaluated by mechanical tests. Bone microarchitecture was evaluated by scanning electron microscopy. One-way ANOVA was used to compare groups. Significance was determined as P<0.05. Regarding bone mineral density, mechanical properties and bone microarchitecture, the beneficial effects were greater in the bones of animals subjected to pre-suspension training and subsequently to training during suspension, compared with the bones of animals subjected to pre-suspension training or to training during suspension. Our results indicate that a period of high impact exercise prior to tail suspension in rats can prevent the installation of osteopenia if there is also training during the tail suspension.

Cell wall polysaccharides of common beans (Phaseolus vulgaris L.)

The soluble and insoluble cotyledon (SPF-Co and IPF-Co) and tegument (SPF-Te and IPF-Te) cell wall polymer fractions of common beans (Phaseolus vulgaris) were isolated using a chemical-enzymatic method. The sugar composition showed that SPF-Co was constituted of 38.6% arabinose, 23.4% uronic acids, 12.7% galactose, 11.2% xylose, 6.4% mannose and 6.1% glucose, probably derived from slightly branched and weakly bound polymers. The IPF-Co was fractionated with chelating agent (CDTA) and with increasing concentrations of NaOH. The bulk of the cell wall polymers (29.4%) were extracted with 4.0M NaOH and this fraction contained mainly arabinose (55.0%), uronic acid (18.9%), glucose (10.7%), xylose (10.3%) and galactose (3.4%). About 8.7% and 10.6% of the polymers were solubilised with CDTA and 0.01M NaOH respectively and were constituted of arabinose (52.0 and 45.9%), uronic acids (25.8 and 29.8%), xylose (9.6 and 10.2%), galactose (6.1 and 3.9%) and glucose (6.5 and 3.8%). The cell wall polymers were also constituted of small amounts (5.6 and 7.2%) of cellulose (CEL) and of non-extractable cell wall polymers (NECW). About 16.8% and 17.2% of the polymers were solubilised with 0.5 and 1.0M NaOH and contained, respectively, 92.1 and 90.7% of glucose derived from starch (IST). The neutral sugar and polymers solubilization profiles showed that weakly bound pectins are present mainly in SPF-Co (water-soluble), CDTA and 0.01-0.1M NaOH soluble fractions. Less soluble, highly cross-linked pectins were solubilised with 4.0M NaOH. This pectin is arabinose-rich, probably highly branched and has a higher molecular weight than the pectin present in SPF-Co, CDTA and 0.01-0.1M NaOH fractions.

Safety and outcome of coronary interventions with special reference to anticoagulation and stent type

University of Turku, Faculty of Medicine, Department of Cardiology and Cardiovascular Medicine, Doctoral Programme of Clinical Investigation, Heart Center, Turku University Hospital, Turku, Finland Division of Internal Medicine, Department of Cardiology, Seinäjoki Central Hospital, Seinäjoki, Finland Heart Center, Satakunta Central Hospital, Pori, Finland Annales Universitatis Turkuensis Painosalama Oy, Turku, Finland 2015 Antithrombotic therapy during and after coronary procedures always entails the challenging establishment of a balance between bleeding and thrombotic complications. It has been generally recommended to patients on long-term warfarin therapy to discontinue warfarin a few days prior to elective coronary angiography or intervention to prevent bleeding complications. Bridging therapy with heparin is recommended for patients at an increased risk of thromboembolism who require the interruption of anticoagulation for elective surgery or an invasive procedure. In study I, consecutive patients on warfarin therapy referred for diagnostic coronary angiography were compared to control patients with a similar disease presentation without warfarin. The strategy of performing coronary angiography during uninterrupted therapeutic warfarin anticoagulation appeared to be a relatively safe alternative to bridging therapy, if the international normalized ratio level was not on a supratherapeutic level. In-stent restenosis remains an important reason for failure of long-term success after a percutaneous coronary intervention (PCI). Drug-eluting stents (DES) reduce the problem of restenosis inherent to bare metal stents (BMS). However, a longer delay in arterial healing may extend the risk of stent thrombosis (ST) far beyond 30 days after the DES implantation. Early discontinuation of antiplatelet therapy has been the most important predisposing factor for ST. In study II, patients on long-term oral anticoagulant (OAC) underwent DES or BMS stenting with a median of 3.5 years’follow-up. The selective use of DESs with a short triple therapy seemed to be safe in OAC patients, since late STs were rare even without long clopidogrel treatment. Major bleeding and cardiac events were common in this patient group irrespective of stent type. In order to help to predict the bleeding risk in patients on OAC, several different bleeding risk scorings have been developed. Risk scoring systems have also been used also in the setting of patients undergoing a PCI. In study III, the predictive value of an outpatient bleeding risk index (OBRI) to identify patients at high risk of bleeding was analysed. The bleeding risk seemed not to modify periprocedural or long-term treatment choices in patients on OAC after a percutaneous coronary intervention. Patients with a high OBRI often had major bleeding episodes, and the OBRI may be suitable for risk evaluation in this patient group. Optical coherence tomography (OCT) is a novel technology for imaging intravascular coronary arteries. OCT is a light-based imaging modality that enables a 12–18 µm tissue axial resolution to visualize plaques in the vessel, possible dissections and thrombi as well as, stent strut appositions and coverage, and to measure the vessel lumen and lesions. In study IV, 30 days after titanium-nitride-oxide (TITANOX)-coated stent implantation, the binary stent strut coverage was satisfactory and the prevalence of malapposed struts was low as evaluated by OCT. Long-term clinical events in patients treated with (TITANOX)-coated bio-active stents (BAS) and paclitaxel-eluting stents (PES) in routine clinical practice were examined in study V. At the 3-year follow-up, BAS resulted in better long-term outcome when compared with PES with an infrequent need for target vessel revascularization. Keywords: anticoagulation, restenosis, thrombosis, bleeding, optical coherence tomography, titanium

Cultivar, harvest year, and storage conditions affecting nutritional quality of common beans (Phaseolus vulgaris L.

Sixteen common bean cultivars were compared concerning the physicochemical characteristics of their raw seeds in the course of two consecutive harvests, as well as the effect of storage conditions on starch and dietary fiber content of cooked beans. Using cluster analysis it was possible to identify groups of cultivars with different nutritional features. Bean cultivars were categorized into four different groups according either to their macronutrient content (crude protein-PROT, total dietary fiber-TDF, insoluble dietary fiber-IDF, soluble dietary fiber-SDF, digestible starch-DS, and resistant starch-RS) or to their micronutrient levels (Fe, Zn, Mn, Cu, Ca, Mg, and P). Guateian 6662 and Rio Tibagi appeared to be the most suitable cultivars to prevent nutritional deficiencies, because they had high PROT, DS, Fe, and Zn content. The high total dietary fiber and RS content of Iraí, Minuano, and TPS Bonito cultivars, and specially the high soluble fiber content of Guateian 6662 and Rio Tibagi cultivars suggests that they could have a beneficial role in controlling blood lipid and glucose levels. Cooked beans had a decrease in DS and an increase in RS content after storage (4 °C or -20 °C), but these changes were more prominent in beans that had low RS content before cooking than in those of high RS content. TDF, IDF, and SDF did not change after storage.

Effect of holding period prior to storage on the chemical attributes of Starking Delicious apples during refrigerated storage

In this research, the effects of three different holding periods (6, 12 and 24 hours) prior to storage on the quality attributes of Starking Delicious apples were investigated during storage of 8 months at 0.5 ± 1.0 ºC. Changes in weight loss, flesh firmness, pH values, soluble dry matter amount, titratable acidity values, ascorbic acid contents, and total and reducing sugar content were determined. According to the results, the holding period showed statistically significant changes in the quality attributes of the apples (p < 0.05).

Nutritional characteristics of biofortified common beans

Iron and zinc deficiency can cause anemia and alterations in the immune response and impair work capacity. To minimize this problem, biofortification has been developed to improve and/or maintain the nutritional status of the population. Beans are an important source of carbohydrates, proteins, and minerals. The objective of this study is to characterize biofortified beans, quantify the minerals in different cultivars, and determine mineral dialysis. Grains of raw and cooked beans were analyzed for moisture, protein, lipids, fiber, minerals, and in vitro availability using four treatments and one control. The data were analyzed using ANOVA, and the Tukey test (p<0.05). The chemical composition of the raw and cooked treatments showed a moisture content ranging from 13.4 to 81.4%, protein from 22.24 to 31.59%, lipids from 1.66 to 2.22%, fiber from 16.81 to 40.63%, carbohydrates from 27.80 to 34.78%, and ash from 4.1 to 4.82%. Different varieties of beans showed statistically significant differences in iron and zinc content compared to the control cultivar (Pérola). The iron content differed significantly from that of the Pérola cultivar in the raw treatment, while in the cooked treatment, the control cultivar did not differ from the Piratã. The same behavior was observed for the zinc content in both treatments. There was no significant difference between the cultivars in the treatments in terms of the content of the dialysis of Calcium (Ca), Iron (Fe), Magnesium (Mg), and Zinc (Zn).

Prevention of contrast induced nephropathy with sodium bicarbonate (the PROMEC study)

Introduction: Contrast-induced nephropathy is a common complication of radiographic procedures. Different measures have been used to avoid this damage, but the evidence is controversial. New investigations are required to clarify it. We investigated the efficacy and safety of sodium bicarbonate solution compared with sodium chloride solution to prevent contrast induced nephropathy in patients with or at risk of renal dysfunction. Methods: A prospective, single-center, randomized clinical trial conducted from May 1, 2007 to February 8, 2008. Inpatients in a tertiary center, scheduled to undergo a procedure with the nonionic radiographic contrast agent iohexol. There were 220 patients with serum creatinine levels of at least 1.2 mg/dL (106.1 µmol/L) and/or type 2 diabetics, who were randomized to receive an infusion of sodium chloride (n = 113) or sodium bicarbonate (n = 107) before and after contrast dye administration. The intervention were "A" group received 1 ml/kg/hour of normal saline solution, starting 12 hours before and continuing 12 hours after iohexol contrast. "B" group received 3 ml/kg of sodium bicarbonate solution (150 mEq/L) one hour prior to procedure and then drip rate was decreased to 1 ml/kg/hour until 6 hours post procedure. Our main outcome measure was change in serum creatinine. Results: The mean creatinine value after the procedure was 1.26 mg/dL in the saline group and 1.22 mg/dL in the bicarbonate group (mean difference: 0.036; CI 95%: -0.16 to 0.23, p = 0.865). The diagnosis of contrast-induced nephropathy, defined by increase in serum creatinine on 25% or more within 2 days after administration of radiographic contrast, was done in twelve patients (12%) in the bicarbonate group and eighth patients (7.1%) in the saline group (RR: 1.68, CI 95%: 0.72 to 3.94). Conclusion: Our investigation showed that there were no differences between normal saline solution (extended infusion) vs. bicarbonate solution for nephroprotection.