An improved protocol for efficient transformation and regeneration of Setaria italica.

KEY MESSAGE: An efficient and improved transformation method for functional genetics studies in S. italica, being a boon for the Setaria scientific community and for crop improvement. Foxtail millet (Setaria italica) is a short life cycle C4 plant, with sequenced genome, and a potential model plant for C4 species. S. italica is also important on a global food security and healthiness context due to its importance in arid and semi-arid areas. However, despite its importance, there are just few transformation protocols directed to this species. The current protocols reached about 5.5-9% of efficiency, which do not make it a valuable model organism. Different types of explants were used in the above mentioned methods, such as immature and mature inflorescence and shoot apex. However, these techniques have many limitations, such as unavailability of explants throughout the year and a crucial, laborious and considerable time-consuming selection. Aiming a simplified and efficient methodology, we adopted dry mature seeds as explants, which are available in abundance, are constant along the year and well responsive to tissue culture, in addition to a differentiated approach that reaches on an average 19.2% transformation efficiency of S. italica. Thus, we propose a protocol that optimizes the transformation efficiency of this cereal crop allowing a high increase on transformation and regeneration rates. Our transformation protocol provides an interesting tool for Setaria community research as well as enables new strategies for breeding enhanced productivity in the species.

Highly sensitive thermoelectric touch sensor based on p-type SnO x thin film.

Here, the ability of using p-type tin oxide (SnO x ) thin films as a thermal sensor has been investigated. Firstly, the thermoelectric performance was optimized by controlling the thickness of the SnO x film from 60 up to 160 nm. A high Seebeck coefficient of +263 µV K-1 and electrical conductivity of 4.1 × 102 (S m-1) were achieved in a 60 nm thick SnO x film, due to a compact nanostructured film and the absence of the Sn metallic phase, which was observed for the thicker SnO x film leading to a typical thermoelectric transport properties of a n-type Sn film. Moreover, x-ray photoelectron spectroscopy revealed the co-existence of SnO (79.7%) and SnO2 (20.3%) phases in the 60 nm thick SnO x film, while the optical measurements revealed an indirect gap of 1.8 eV and a direct gap of 2.7 eV, respectively. The 60 nm-SnO x thin film have been tested as a thermoelectric touch sensor, achieving a Vsignal /Vnoise  ≈ 20, with a rise time <1 s. Therefore, this work provides an efficient way for developing highly efficient thermal sensors with potential use in display technologies.

Association of CXCL13 serum level and ultrasonographic findings of joints in patients with systemic lupus erythematosus and Jaccoud's arthropathy.

Objectives The objective of this paper is to perform an ultrasonography (US) analysis of hands and wrists in two groups of patients with systemic lupus erythematosus (SLE), with and without Jaccoud's arthropathy, matched by age and disease duration and to correlate them with levels of CXCL13 clinical features, laboratory tests and disease activity score. Methods Sixty-four patients with SLE were enrolled, 32 with and 32 without Jaccoud's arthropathy. Each patient underwent physical examination, laboratory tests (including CXCL13 by ELISA) and bilateral US. Synovial hypertrophy, tenosynovitis and erosions were evaluated according to a semiquantitative grading system with a 0-3 rating. US findings were correlated with serum levels of CXCL13, other serological parameters and disease activity index. Results Synovitis was found in 25/64 patients (39%) and tenosynovitis in 14/64 (22%). These findings were more frequent in SLE patients with Jaccoud's arthropathy, particularly tenosynovitis ( p = 0.002) and synovitis ( p = 0.01). Median serum level of CXCL13 was 20.16 pg/ml in the whole population (23.21 pg/ml in the Jaccoud's arthropathy group and 11.48 pg/ml in the group without). There was an association between the presence of disease activity and high level of CXCL13 ( p = 0.004). However, no association was found between high levels of CXCL13 and "arthritis" in SLEDAI, swollen joints on physical examination or synovitis on US. Conclusions US findings in joints of SLE patients with Jaccoud's arthropathy confirm that synovitis and tenosynovitis are common in these patients. In addition, serum level of CXCL13 is associated with disease activity in SLE but does not seem to be a biomarker for arthritis in these patients.

Bodybuilding protein supplements and cow's milk allergy in adult.

Summary: We report a case of a previously healthy 24-year-old man with a 3-month history of gastrointestinal symptoms during exercise and also few minutes after the ingestion of cow's milk (CM) without exercise. He reported the ingestion of a blend of hydrolyzed whey and casein proteins for bodybuilding for the last 2 years. The in vivo tests showed positivity to CM, α-lactalbumin, ß-lactoglobulin and casein extracts, and also to the protein supplement. The serum specific IgE was positive for CM, ß-lactoglobulin and α-lactalbumin. The in vivo and in vitro tests results suggested an IgE-mediated CMA. Adult-onset CMA has been rarely reported, and to our knowledge this is the first case possibly related to bodybuilding supplements. The authors theorize that the presentation of large amounts of proteins in the gastrointestinal tract may favor sensitization.

Localized salt-dependent aquagenic urticaria, a rare subtype of urticaria: a case report.

Summary: Aquagenic urticaria (AU) is a rare form of chronic inducible urticaria elicited by water at any temperature. We describe the case of a 25-year-old atopic woman who presented to our unit with a 4-year history of recurrent urticarial rashes, highly pruritic, confined to the neck and lower part of the face, occurring solely on contact with sea water. The lesions were reproduced by challenge tests with aqueous 3.5% NaCl and other hypertonic aqueous solutions but not with 20% glucose neither tap water. Our case supports the existence of a distinct salt-dependent subtype of aquagenic urticaria (SDAU), which seems to be triggered mostly by sea bathing, affects young women and has a characteristic localization on the inferior facial contours and neck. To the best of our knowledge, only eight cases of SDAU have been reported in the literature.

Hypersensitivity to antiretroviral drugs.

Summary: Background. Antiretroviral therapy (ART) may be responsible for hypersensitivity reactions varying in severity, clinical manifestations and frequency. Case report. We report the case of a 47-year-old woman with HIV infection who developed a delayed mucocutaneous reaction after treatment with ART. Hypersensitivty reaction (HR) to emtricitabine and tenofovir was considered probable based on positive patch tests (PT) and hypersensitivity reaction to nevirapine was confirmed by drug provocation test. Discussion. The diagnosis of HR to ART remains a diagnostic challenge, partly due to unknown mechanism and the absence of validated diagnostic tools. Patch testing may represent a useful method for confirming hypersensitivity. Further investigation in this area is required, so that successful management strategies can be offered, preventing loss of potent and viable antiretroviral agents.

Coupled transcript and metabolite identification: insights on induction and synthesis of resveratrol in peanut, wild relatives and synthetic allotetraploid.

Resveratrol is an antioxidant that is a promising antitumoral, cardioprotective and neuroprotective agent. It has been found in a restricted number of plants including peanut (Arachis hypogaea L.) and its wild relatives. The objective of this study was to understand the relationship between resveratrol content and the expression of putative resveratrol synthase genes in four Arachis genotypes. Two diploids and two tetraploid were analyzed. Contents of resveratrol on non- and UV-treated leaves were estimated using HPLC. Resveratrol synthase (RS) was analyzed using RT-qPCR with primers developed in this study. Sequences of six Arachis species were amplified using two degenerated primer pairs that were designed based on Arachis and general RS available at GenBank. Those sequences were used to qPCR primers design. Test and control leaves were collected from plants cultivated in greenhouse and three biological replicates were evaluated for each genotype. The synthesis of resveratrol in leaves was induced by treatment with UV for 2.5 h. All genotypes studied synthesized resveratrol. Concentrations ranged from 193.66 µg/g in synthetic allotetraploid to 371.97 µg/g in A. duranensis. Natural and induced allotetraoploids showed lower levels of resveratrol than their diploid parents. Untreated samples did not produce significant amounts of resveratrol. The analysis of resveratrol content and levels of RS mRNA allowed the identification of one gene induced by the UV treatment. The data showed different amounts of RS in the different genotypes suggesting early and late response to the UV induction in the different species. The understanding of the variation found among species will help to identify species that have high resveratrol content and their ideal pos-induction times. This also will allow analysis of other tissues where high levels resveratrol would be very important, such as in seeds.

Design, synthesis, molecular docking and biological evaluation of thiophen-2-iminothiazolidine derivatives for use against Trypanosoma cruzi.

In this study, we designed and synthesized a series of thiophen-2-iminothiazolidine derivatives from thiophen-2-thioureic with good anti-Trypanosoma cruzi activity. Several of the final compounds displayed remarkable trypanocidal activity. The ability of the new compounds to inhibit the activity of the enzyme cruzain, the major cysteine protease of T. cruzi, was also explored. The compounds 3b, 4b, 8b and 8c were the most active derivatives against amastigote form, with significant IC50 values between 9.7 and 6.03µM. The 8c derivative showed the highest potency against cruzain (IC50=2.4µM). Molecular docking study showed that this compound can interact with subsites S1 and S2 simultaneously, and the negative values for the theoretical energy binding (Eb=-7.39kcal·mol(-1)) indicates interaction (via dipole-dipole) between the hybridized sulfur sp(3) atom at the thiazolidine ring and Gly66. Finally, the results suggest that the thiophen-2-iminothiazolidines synthesized are important lead compounds for the continuing battle against Chagas disease.

Use of omalizumab in the treatment of chronic urticaria.

BACKGROUND: Omalizumab is indicated to treat chronic spontaneous urticaria (CSU) refractory to antihistamines. We aim to describe the experience of our department in the treatment of CSU with omalizumab. MATERIALS AND METHODS: Retrospective review of the clinical records of patients. RESULTS: Six patients (5 females, median age 33 years) treated with omalizumab for a median of 17.5 months were evaluated. All patients had improvement of symptoms after the first dose. In one case, the treatment was suspended after 7 months, but in 9 weeks there was recurrence of symptoms. The main side effect was headache in the drug administration's day. Currently, all patients maintain therapy with omalizumab and are clinically stable. CONCLUSION: Omalizumab proved to be an effective and safe drug for the treatment of patients with refractory CSU.

Response of Soybean Genotypes Challenged by a Stink Bug Complex (Hemiptera: Pentatomidae).

Pentatomids (stink bugs) are major pests of soybean, Glycine max (L.) Merril. These pests reach high levels of infestation, cause severe damage to seeds by feeding, are linked to leaf retention, and are difficult to control. Host plant resistance is considered to be a valuable tool in integrated pest management and can assist in reducing the damage caused by stink bugs. This research evaluated the resistance of soybean genotypes in Brazil to the stink bug complex, the Neotropical brown stink bug, Euschistus heros (F.), redbanded stink bug, Piezodorus guildinii (Westwood), southern green stink bug, Nezara viridula (L.), green belly stink bug, Dichelops melacanthus (Dallas), and Edessa meditabunda (F.), by assessing infestation assay, yield reduction, seed damage, and leaf retention. Certain genotypes expressed different categories of resistance: least infested, low yield reduction, low levels of damage in seeds, and low levels of leaf retention. PI lines and IAC 78-2318 showed antixenotic resistance, and 'IAC 100' showed tolerance for the stink bug complex. This is the first study to evaluate several parameters of yield and seed quality using different soybean maturity groups under relatively high infestation by the three stink bugs species. The promising genotypes might be used in regions with a high incidence of stink bugs to manage their populations in combination with other integrated pest management practices.

Structural dynamics and physicochemical properties of pDNA/DODAB:MO lipoplexes: effect of pH and anionic lipids in inverted non-lamellar phases versus lamellar phases.

Dioctadecyldimethylammonium bromide (DODAB):Monoolein (MO) lipoplexes have mainly been studied within the range of high molar ratios of DODAB, with noticeable transfection efficiencies in the Human Embryonic Kidney (HEK, a.k.a. 293T) cell line. In this work, we intend to study the effect of high MO content on the structure and physicochemical properties of pDNA/DODAB:MO lipoplexes to achieve some correlation with their transfection efficiency. Static/Dynamic Light Scattering and Cryo-TEM imaging were used to characterize the size/morphology of DNA/DODAB:MO lipoplexes at different DODAB:MO contents (2:1, 1:1, 1:2) and charge ratios (CRs) (+/-). Nile Red fluorescence emission was performed to detect changes in microviscosity, hydration and polarity of DNA/DODAB:MO systems. Lipoplexes stability at physiological pH values and in the presence of anionic lipids was evaluated by Förster Resonance Energy Transfer (FRET). Physicochemical/structural data were complemented with transfection studies in HEK cells using the ß-galactosidase reporter gene activity assay. This work reports the coexistence of multilamellar and non-lamellar inverted phases in MO-richer lipoplexes (DODAB:MO 1:2 and 1:4), leading to transfection efficiencies comparable to those of multilamellar (DODAB-richer) lipoplexes, but at higher charge ratios [CR (+/-)=6.0] and without dose-effect response. These results may be related to the structural changes of lipoplexes promoted by high MO content.

Dextrin-based nanomagnetogel: in vivo biodistribution and stability.

The biodistribution profile of a new dextrin nanomagnetogel, which consists of γ-Fe2O3 superparamagnetic nanoparticles loaded within a polymeric matrix of modified dextrin, was studied in mice. The nanomagnetogel bear a monomodal size distribution profile (average diameter 110 nm) close to neutral surface charge and higher relaxivity (r2 = 215-248 mM(-1) s(-1) and r2/r1 = 13-11) than those of commercial formulations (r2 = 160-177 mM(-1) s(-1) and r2/r1 = 4-7). Also, the observed blood half-life-approximately 4 h-is superior to that of similar commercially available formulations, which remain for a few minutes in circulation. PEGylation resulted in 1.7- and 1.2-fold lower accumulation in the liver and spleen, respectively, within the first 24 h. Noteworthy, a good correlation was obtained between the amount of polymer (quantified by scintigraphy) in the spleen, 48 h after administration, and the amount of iron physically loaded through hydrophobic interactions (quantified by ICP) indicating the absence of iron leakage from the polymeric matrix. This study provides evidence of the in vivo stability of a self-assembled nanomagnetogel, a relevant feature which is seldom reported in the literature.

Multiplicity of steady states in a bioreactor during the production of 1,3-propanediol by Clostridium butyricum.

In recent decades, the production of compounds from microorganisms has increased significantly. Glycerol as a source of substrate appears to have great potential, due to its large supply because of the increase in biodiesel production. This paper will discuss the multiplicity of steady states for the production of 1,3-propanediol from glycerol by Clostridium butyricum, employing a model that takes into account inhibition by fermentation products. The theoretical study of bifurcation enabled us to make a qualitative adjustment to the various experimental steady states, using the theoretical steady states obtained from the AUTO2007 program. The theoretical model parameters were varied to fit qualitatively the values of the experimental steady states. In addition, this work is a qualitative study, using experimental steady states that can be used as an initial study for more advanced work on optimizing the production of 1,3-propanediol.

Antimicrobial and plant growth-promoting properties of the cacao endophyte Bacillus subtilis ALB629.

AIMS: To investigate the effects of the endophyte Bacillus subtilisALB629 on the growth of cacao seedlings at early developmental stage and to evaluate its antimicrobial properties. METHODS AND RESULTS: Germinating cacao seeds were inoculated with ALB629, and seedlings growth was evaluated 30 days later. Significant increase (P < 0·05) was observed in the root system (up to 30%), leaf area (14%) and stem height (7·6%). ALB629 colonized the entire plant, prevailing over indigenous micro-organisms. In addition, it was tested in vitro, by pairing assays, and showed antagonistic effect against the phytopathogenic fungi Moniliophthora perniciosa, Colletotrichum sp. and C. gossypii. When tested in cacao-grafting procedure in the field, ALB629 increased the grafting success rate (24%), indicating its protective effect. In addition, this Bacillus secretes an antagonist compound, as shown by the antifungal activity of the cell-free culture. CONCLUSIONS: Bacillus subtilisALB629 promotes cacao root growth, besides promoting growth of the aerial part of cacao seedlings. It has antimicrobial properties and produces an antifungal compound. SIGNIFICANCE AND IMPACT OF THE STUDY: ALB629 presented beneficial characteristics for cacao cultivation, being a good biological control agent candidate. Furthermore, it is a potential source of antifungal compound with potential for commercial exploitation.

Desensitization to clopidogrel: a tailor-made protocol.

Clopidogrel is an antiplatelet drug widely used for treatment and prevention of a variety of cardiovascular diseases. We report a successful desensitization to clopidogrel in a 70-year-old Caucasian man with delayed hypersensitivity (HS) reaction. He developed lip, hand and foot swelling, erythematous papular non-pruritic lesions and arthralgias 2 weeks after starting treatment with clopidogrel 75 mg/d. A 3-hour desensitization protocol was started, achieving a cumulative dose of 154 mg without any reaction, and a daily dose of 75 mg was recommended. On the 4th day, the patient developed skin lesions similar to the previously described. He was treated with topical steroids and oral antihistamines, and the daily dose of clopidogrel was reduced to 20 mg. A new desensitization protocol was established, with a slow dose increment, according to the patient's response. It was only possible to achieve the dose of 75 mg/d after 2 months. Although well tolerated by most patients, HS reactions with clopidogrel may occur and desensitization is rising as a safe alternative in those patients. In delayed reactions with cutaneous lesions, a slower desensitization protocol may be necessary, as in this case.

Amazonian fermentations: an analysis of industrial and social technology as tools for the development of bioeconomy in the region.

This review article explores the potential of fermentations in the Amazon region as catalysts for economic and social development. It highlights the rich cultural and gastronomic diversity of the Amazon, focusing on indigenous fermented products. Two main products, tucupi and caxiri, are discussed in detail, emphasizing their significance in local cuisine and culture. The review examines the challenges and opportunities for industrial applications of these products, as well as their potential for social technology initiatives, particularly in the context of family farming. The sustainable production of native fermented products in the Amazon is seen as a means to preserve biodiversity, empower local communities, and promote cultural heritage. The article concludes that both industrial and social technologies have complementary roles in promoting economic growth, cultural preservation, and the well-being of the Amazon region, making it a promising hub for innovative and sustainable fermented food products on a global scale.

Case report: A novel TET2 mutation in a patient with refractory cytopenia with multilineage dysplasia.

Myelodysplastic syndrome diagnosis of karyotypically normal patients may be elusive because it relies exclusively on morphological and clinical data. In routine practice, finding of an acquired mutation or a cytogenetic abnormality provides irrefutable evidence of the clonal nature of that disease. Recurrent deletions and somatic mutations in TET2, a gene involved in epigenetic regulation, have been reported in about 20% of adult patients with myelodysplastic syndrome. We report a novel g.95805C>T, nonsense TET2 mutation, leading to a premature stop codon (p.Gln913*), in an adult patient with refractory cytopenia with multilineage dysplasia.

Assessment of asthma control: clinical, functional and inflammatory aspects.

BACKGROUND: Asthma is a complex disease with numerous markers of severity/activity. Clinical assessment,functional parameters and inflammation biomarkers are the most used A correlation between them is difficult, as each one evaluates a particular aspect of the disease. OBJECTIVE AND METHODS: To explore the possible association between asthma control, pulmonary function and inflammation in patients with asthma, consecutive asthmatics underwent simultaneous spirometry (measurement of FEV1), exhaled nitric oxide (eNO) evaluation and Asthma Control Test (ACTTM) questionnaire. RESULTS: The study included 232 asthmatics (mean age: 37.48 years; 78.4%female): 43% had uncontrolled asthma (ACTTM < or = 19) with FEV1 mean values of 83.3% +/- 21.8; 48% partially controlled (ACTTM: 20-24) with FEV1 of 87.6% +/- 17; 9% complete control (ACTTM = 25) with FEV1 of 93.1 +/- 20.6. The relation ACTTM/FEV1 and ACTTM/FEF 25-75% was statistically significant (p = 0.001 and p = 0.034, respectively). Among patients with eNO < 35 ppb, 66% had FEV1 > 80% and 52% had ACTTM > 19. No association was found combining ACTTM/eNO or FEV1/eNO. A subgroup of 66 patients was evaluated twice. CONCLUSION: An association was found between ACTTM and spirometry, with higher ACTTM scores reflecting less bronchial obstruction. The authors advise a combined approach in asthma follow-up, involving clinical aspects, functional parameters and inflammation biomarkers, although in some circumstances ACT could be a valid instrument to be used alone to assess control.

Impaired insulin secretion and beta-cell loss in tissue-specific knockout mice with mitochondrial diabetes.

Mitochondrial dysfunction is an important contributor to human pathology and it is estimated that mutations of mitochondrial DNA (mtDNA) cause approximately 0.5-1% of all types of diabetes mellitus. We have generated a mouse model for mitochondrial diabetes by tissue-specific disruption of the nuclear gene encoding mitochondrial transcription factor A (Tfam, previously mtTFA; ref. 7) in pancreatic beta-cells. This transcriptional activator is imported to mitochondria, where it is essential for mtDNA expression and maintenance. The Tfam-mutant mice developed diabetes from the age of approximately 5 weeks and displayed severe mtDNA depletion, deficient oxidative phosphorylation and abnormal appearing mitochondria in islets at the ages of 7-9 weeks. We performed physiological studies of beta-cell stimulus-secretion coupling in islets isolated from 7-9-week-old mutant mice and found reduced hyperpolarization of the mitochondrial membrane potential, impaired Ca(2+)-signalling and lowered insulin release in response to glucose stimulation. We observed reduced beta-cell mass in older mutants. Our findings identify two phases in the pathogenesis of mitochondrial diabetes; mutant beta-cells initially display reduced stimulus-secretion coupling, later followed by beta-cell loss. This animal model reproduces the beta-cell pathology of human mitochondrial diabetes and provides genetic evidence for a critical role of the respiratory chain in insulin secretion.

Note on the measurement of bulk density and tapped density of powders according to the European Pharmacopeia.

The apparent volume and compressibility index of commonly used excipients were evaluated according to European Pharmacopeia monograph (seventh edition) in order to study the influence of the procedure conditions. The results suggested that the leveling of the powder inside the cylinder ought to be avoided.