Gelatin-Chitosan Hydrogel Biological, Antimicrobial and Mechanical Properties for Dental Applications.

Chitosan, a natural polysaccharide sourced from crustaceans and insects, is often used with hydrogels in wound care. Evaluating its cytotoxicity and antimicrobial properties is crucial for its potential use in dentistry. OBJECTIVE: To investigate the mechanical properties of gelatin hydrogels based on decaethylated chitosan and antimicrobial activity against Streptococcus mutans and their biological effects with stem cells from apical papilla (SCAPs). MATERIAL AND METHODS: Gelatin-chitosan hydrogels were synthesized at concentrations of 0%, 0.2% and 0.5%. Enzymatic and hydrolytic degradation, along with swelling capacity, was assessed. Fourier transform infrared spectroscopy (FTIR) analysis was employed to characterize the hydrogels. The interaction between hydrogels and SCAPs was examined through initial adhesion and cell proliferation at 24 and 48 h, using the Thiazolyl Blue Tetrazolium Bromide (MTT assay). The antimicrobial effect was evaluated using agar diffusion and a microdilution test against S. mutans. Uniaxial tensile strength (UTS) was also measured to assess the mechanical properties of the hydrogels. RESULTS: The hydrogels underwent hydrolytic and enzymatic degradation at 30, 220, 300 min and 15, 25, 30 min, respectively. Significantly, (p < 0.01) swelling capacity occurred at 20, 40, 30 min, respectively. Gelatin-chitosan hydrogels' functional groups were confirmed using vibrational pattern analysis. SCAPs proliferation corresponded to 24 h = 73 ± 2%, 82 ± 2%, 61 ± 6% and 48 h = 83 ± 11%, 86 ± 2%, 44 ± 2%, respectively. The bacterial survival of hydrogel interaction was found to be 96 ± 1%, 17 ± 1.5% (p < 0.01) and 1 ± 0.5% (p < 0.01), respectively. UTS showed enhanced (p < 0.05) mechanical properties with chitosan presence. CONCLUSION: Gelatin-chitosan hydrogels displayed favorable degradation, swelling capacity, mild dose-dependent cytotoxicity, significant proliferation with stem cells from apical papilla (SCAPs), substantial antimicrobial effects against S. mutans and enhanced mechanical properties. These findings highlight their potential applications as postoperative care dressings.

Decellularized Scaffolds of Nopal (Opuntia Ficus-indica) for Bioengineering in Regenerative Dentistry.

Opuntia Ficus-indica, or nopal, is traditionally used for its medicinal properties in Mexico. This study aims to decellularize and characterize nopal (Opuntia Ficus-indica) scaffolds, assess their degradation and the proliferation of hDPSC, and determine potential pro-inflammatory effects by assessing the expression of cyclooxygenase 1 and 2 (COX-1 and 2). The scaffolds were decellularized using a 0.5% sodium dodecyl sulfate (SDS) solution and confirmed by color, optical microscopy, and SEM. The degradation rates and mechanical properties of the scaffolds were determined by weight and solution absorbances using trypsin and PBS and tensile strength testing. Human dental pulp stem cells (hDPSCs) primary cells were used for scaffold-cell interaction and proliferation assays, as well as an MTT assay to determine proliferation. Proinflammatory protein expression of COX-I and -II was discovered by Western blot assay, and the cultures were induced into a pro-inflammatory state with interleukin 1-ß. The nopal scaffolds exhibited a porous structure with an average pore size of 252 ± 77 µm. The decellularized scaffolds showed a 57% reduction in weight loss during hydrolytic degradation and a 70% reduction during enzymatic degradation. There was no difference in tensile strengths between native and decellularized scaffolds (12.5 ± 1 and 11.8 ± 0.5 MPa). Furthermore, hDPSCs showed a significant increase in cell viability of 95% and 106% at 168 h for native and decellularized scaffolds, respectively. The combination of the scaffold and hDPSCs did not cause an increase in the expression of COX-1 and COX-2 proteins. However, when the combination was exposed to IL-1ß, there was an increase in the expression of COX-2. This study demonstrates the potential application of nopal scaffolds in tissue engineering and regenerative medicine or dentistry, owing to their structural characteristics, degradation properties, mechanical properties, ability to induce cell proliferation, and lack of enhancement of pro-inflammatory cytokines.

Design of a low-density SNP panel for intramuscular fat content and fatty acid composition of backfat in free-range Iberian pigs.

Intramuscular fat (IMF) content and fatty acids (FA) profile determine important sensory and technological characteristics of the meat, being interesting traits to be included in breeding programs for Iberian pig. In a previous study, we analyzed the longissimus dorsi transcriptome of Iberian pigs with divergent breeding values for IMF identifying 63 and 12 functional candidate genes and long non-coding RNA being differentially expressed. The first objective of this work was to use the RNA-seq data generated in that study in order to design a low-density panel with 50 single-nucleotide polymorphisms (SNP). The second aim was to evaluate the effects of this SNP panel on IMF content and backfat FA profile in a closed commercial population of Iberian pigs. The IMF content and backfat FA composition were determined in 940 Iberian pigs. A total of 837 SNPs were identified in the selection of genes and lncRNA. Out of the 50 selected SNP, 11 were monomorphic in the population and 2 failed in their genotyping. Association analyses were carried out for the 37 SNP segregating in the population (minor allele frequency, MAF > 0.05). Regarding IMF content, a significant additive effect (q-value < 0.05) was observed for ADIPOQ:g.124646194T > G SNP, in which the G allele was linked to an increase of 7.59% (additive effect expressed as a percentage of the mean of the trait). For the backfat FA composition, we observed 24 SNP with significant associations. The strongest effects were detected for the ELOVL6:g.112186423A > G and FASN_rs331694510G > A SNP. The A allele of both SNPs were associated to a lower percentage of palmitic (C16:0) and palmitoleic (C16:1) acids and an increase in oleic acid (C18:1). In addition, the AMPD3:g.49043886C > G SNP had significant effects on C16:0, stearic (C18:0), C18:1 and on the sum of saturated and monounsaturated fatty acids. Finally, significant effects of the EGR2:66286345G > T SNP on C18:0 and C18:1 and other minor FA were observed. Our findings reveal ADIPOQ:g.124646194T > G, ELOVL6:g.112186423A > G, and FASN_rs331694510G > A may be useful in breeding programs for Iberian pigs including FA profile and IMF content as selection goals, provided that their potential effects on other traits of interest are controlled.

Fat infiltration of muscle and fatty acid profile of cover fat are essential traits in Iberian pig industry. However, their inclusion in genetic selection schemes can be tangled, since their determination is laborious and expensive. Molecular markers can be a useful alternative to improve these traits. In a previous study, we analyzed the longissimus dorsi transcriptome of Iberian pigs with divergent breeding values for intramuscular fat (IMF) identifying 63 and 12 functional candidate genes and long non-coding RNA being differentially expressed. From this information, we designed a low-density panel with 50 single-nucleotide polymorphisms, which we subsequently evaluated for IMF content and fatty acids (FA) backfat profile of 940 pigs. The present research identified five polymorphisms mapped to ADIPOQ, ELOVL6, FASN, AMPD3, and EGR2 genes that are associated to IMF content and backfat FA profile.

Effects of Nitrurized Titanium on Microhardness and Human Dental Pulp Stem Cell Adhesion and Differentiation.

To compare the Vickers microhardness, surface roughness, initial adhesion, and osteogenic differentiation on titanium (Ti) and nitrurized titanium (NTi) plates were treated by UV irradiation and chitosan. Each plate was subjected to Vickers hardness with a pressure of 2.9 N for 10 seconds and roughness evaluation by atomic force microscope (AFM) analysis. Three groups of each type of plates were tested: control (C), ultraviolet irradiation (UV), and chitosan (Q). The UV group was exposed to UV-irradiation for 20 min at 253.7 nm (52 µW/cm2). The Q group was coated with 1% chitosan, and the C group had no treatment. The osteoblasts (2 × 106 cells/mL) were inoculated in each group for 60 min and their viability was determined by the MTT bioassay. Osteogenic differentiation was performed over 4 weeks and determined by alizarin red staining. The mean was analyzed with the Shapiro-Wilks, Kruskall-Wallis, and Mann-Whitney U tests of normality (n = 9/gp). The NTi plates hardness (125.1 ± 4.01 HV) was higher (P = 0.026) than the Ti plates (121.3 ± 2.23 HV). The surface topography was: NTi (Ra = 0.098 µm) and Ti (Ra = 0.212 µm). The quantification of cell adhesion was: Ti + Q = 123 ± 4.9% (P < 0.05) < NTi + Q = 107 ± 3.3% < Ti = 100 ± 10.7% < NTi = 72 ± 6.8% < NTi + UV = 71 ± 4.4% < Ti + UV = 69 ± 3.5%, regardless the plates, the presence of chitosan induce a faster osteogenic differentiation. The Ti + Q plates tested the highest cell attachment and osteogenic adhesion suggesting their potential use of chitosan for cell-implant interaction.

Natural Bioactive Epigallocatechin-Gallate Promote Bond Strength and Differentiation of Odontoblast-like Cells.

The (-)-Epigallocatechin-gallate (EGCG) metabolite is a natural polyphenol derived from green tea and is associated with antioxidant, biocompatible, and anti-inflammatory effects. OBJECTIVE: To evaluate the effects of EGCG to promote the odontoblast-like cells differentiated from human dental pulp stem cells (hDPSCs); the antimicrobial effects on Escherichia coli, Streptococcus mutans, and Staphylococcus aureus; and improve the adhesion on enamel and dentin by shear bond strength (SBS) and the adhesive remnant index (ARI). MATERIAL AND METHODS: hDSPCs were isolated from pulp tissue and immunologically characterized. EEGC dose-response viability was calculated by MTT assay. Odontoblast-like cells were differentiated from hDPSCs and tested for mineral deposition activity by alizarin red, Von Kossa, and collagen/vimentin staining. Antimicrobial assays were performed in the microdilution test. Demineralization of enamel and dentin in teeth was performed, and the adhesion was conducted by incorporating EGCG in an adhesive system and testing with SBS-ARI. The data were analyzed with normalized Shapiro-Wilks test and ANOVA post hoc Tukey test. RESULTS: The hDPSCs were positive to CD105, CD90, and vimentin and negative to CD34. EGCG (3.12 µg/mL) accelerated the differentiation of odontoblast-like cells. Streptococcus mutans exhibited the highest susceptibility < Staphylococcus aureus < Escherichia coli. EGCG increased (p < 0.05) the dentin adhesion, and cohesive failure was the most frequent. CONCLUSION: (-)-Epigallocatechin-gallate is nontoxic, promotes differentiation into odontoblast-like cells, possesses an antibacterial effect, and increases dentin adhesion.

Fracture Resistance of Endodontically Restored Teeth with Two System Root Post

ABSTRACT: The objective of this study was to determinate the fracture resistance of endodontically treated teeth and restored with two root post systems: i) resin post, ii) fiber post. A total of 60 teeth were freshly extracted, endodontic ally treated and randomly divided in two groups (n= 30/each group) for standardized restoration; Group 1 (Group R): Resin post and resin restoration, Group 2 (Group FP): Fiberglass post and resin restoration. Both groups' samples were mounted in a metallic base at 135º to allow them to be stabilized and held in the universal testing machine by applying a vertical force at cross speed of 1mm/min. Data were recorded in Newtons (N) Previous to test the fracture resistance; all samples were stored in distilled water at 37 ºC for 24 hours. Data were subject to the Saphiro-Wilk test for normality distribution and Student's t test. Significance was considered at 0.05 values. The values of fiber post group showed normal distribution compared to the resin group, demonstrating less variability among the values. The group FP displayed higher fracture resistance (299.77±100 N) than group R (205.57±86.40 N), with significant differences (p= 0.00002). The greatest fracture resistance was recorded for the group having fiber post reinforced and composite cores. It is suggested that fiberglass post restoration is the first option when endodontic treatment requires core restoration.

RESUMEN: El objetivo del estudio fue determinar la resistencia a la fractura de dientes tratados endodónticamente y restaurados con dos sistemas de endopostes radiculares: I) poste de resina, II) poste de fibra. Un total de 60 dientes recién extraídos fueron tratados endodónticamente y divididos al azar en dos grupos (n= 30/ cada grupo) para la restauración estandarizada; Grupo 1 (Grupo R): Pilares de resina y restauración, Grupo 2 (Grupo FP): Pilares de fibra de vidrio y restauración de resina. Las muestras de ambos grupos se montaron en una base metálica a 135º para permitir su estabilización y sujeción en la máquina universal de ensayos aplicando una fuerza vertical a velocidad transversal de 1 mm/min. Los datos se registraron en Newtons (N), para probar la resistencia a la fractura; todas las muestras se almacenaron en agua destilada a 37 ºC durante 24 horas. Los datos se sometieron a la prueba de normalidad de Saphiro-Wilk y la pruebas t de Student. La significancia se consideró con un valor de 0,05. Los valores del grupo de postes de fibra mostraron una distribución normal en comparación con el grupo de resinas, demostrando menor variabilidad entre los valores. El grupo FP mostró mayor resistencia a la fractura (299,77±100 N) que el grupo R (205,57±86,40 N) que el grupo con diferencias significativas (p= 0,00002). La mayor resistencia a la fractura se registró para el grupo que tenía núcleos compuestos y reforzados con postes de fibra. Se sugiere que la restauración posterior de fibra de vidrio es la primera opción cuando el tratamiento de endodoncia requiere una restauración del núcleo.

CRISPR/Cas gene-editing technology and its advances in dentistry.

A recent discovery of revolutionary Clustered regularly interspaced palindromic repeats (CRISPR) is a gene-editing tool that provides a type of adaptive immunity in prokaryotic organisms, which is currently used as a revolutionizing tool in biomedical research. It has a mechanism of correcting genome errors, turning on/off genes in cells and organisms. Most importantly playing a crucial function in bacterial defence by identifying and destroying Deoxyribonucleic acid (DNA) segments during bacteriophage invasions since the CRISPR-associated protein 9 (Cas9) enzyme recognizes and cleaves invasive DNA sequences complementary to CRISPR. Therefore, researchers employ this biological device to manipulate the genes to develop new therapies to combat systemic diseases. Currently, the most significant advance at the laboratory level is the generation of cell and animal models, functional genomic screens, live images of the cell genome, and defective DNA repairs to find the cure for genetic disorders. Even though this technology has enormous biomedical applications in various sectors, this review will summarize CRISPR/Cas emphasizing both the therapeutic and diagnostic mechanisms developed in the field of dentistry and the promising attempts to transfer this technology to clinical application. Finally, future developments are also described, which proposes to use CRISPR/Cas systems for prospective clinical dentistry applications.

Genetic parameter estimation and gene association analyses for meat quality traits in open-air free-range Iberian pigs.

Meat quality of Iberian pigs is defined by the combination of their genetic characteristics and the particular production system. To carry out a genetic analysis of the main meat quality traits, we estimated their heritabilities, genetic correlations and the association effects of 32 selected SNPs of 12 candidate genes. A total of ten traits were measured in longissimus dorsi samples from 1,199 Iberian pigs fattened in the traditional free-range system: water holding capacity (thawing, cooking and centrifuge force water losses), instrumental colour (lightness L*, redness a* and yellowness b*), myoglobin content, shear force on cooked meat, and shear force and maximum compression force on dry-cured loin. Estimated heritability values were low to moderate (0.01 to 0.43) being the lowest for L* and the highest for cooking loss. Strong genetic correlations between water holding capacity traits (0.93 to 0.96) and between myoglobin content and a* (0.94) were observed. The association analyses revealed 19 SNPs significantly associated with different traits. Consistent and strong effects were observed between PRKAG3 SNPs (rs319678464G > C and rs330427832C > T), MYH3_rs81437544T > C, CASP3_rs319658214G > T and CTSL_rs332171512A > G and water losses. Also for CAPN1_rs81358667G > A and CASP3_rs319658214G > T and shear force. The SNPs mapping on PRKAG3 showed the highest effects on Minolta colour traits. Genotyping of these SNPs could be useful for the selection of Iberian young boars with similar estimated breeding values for productive traits.

Foraging strategy of a carnivorous-insectivorous raptor species based on prey size, capturability and nutritional components.

Optimal foraging theory has typically paid little attention to species feeding on mobile prey and has emphasised energy intake rather than the nutritional contribution of food. The difficulty of capturing food has rarely been included in foraging models, even when it is a potentially important modulator of time devoted to foraging. From the central place foraging and provisioning perspectives, it is posited that at high levels of prey selectivity, the time spent to capture prey is longer than at low levels of prey selectivity. Furthermore, in the case of carnivorous predators, it is thought that nutritional composition does not influence foraging strategies. To explore these issues, we investigated the influence of abundance, size, difficulty of capture, gross energy and nutritional composition (fat, protein, protein-fat ratio and amino acid contents) of prey species on the foraging behaviour of a predator species, the common kestrel Falco tinnunculus, in a region of high diversity of prey species. Our results show that capturability index and load-size explain the foraging behaviour of kestrels. Preferred prey take longer to be provisioned, both selectivity and capturability might explain this result. It is also shown that specific nutritional components, such as protein and amino acid contents, are likely to explain food preference in this carnivorous-insectivorous species.

Genotype and dietary lysine deficiency affect carcass and muscle amino acid composition of pigs growing from 10 to 25 kg body weight.

Amino acid (AA) composition of body protein is considered constant although there are evidences that AA pattern in pigs may be altered by different factors. Pigs with different body composition and protein deposition rates-like fatty and lean pigs-may differ in AA composition, with possible consequences on their AA requirements. This work investigates effects of genotype and dietary lysine deficiency on AA composition of carcass and muscles of Iberian and Landrace × Large White pigs. Twenty-eight barrows (10 kg body weight [BW]), 14 from each breed, were used. They were randomly assigned to two experimental diets according to a factorial arrangement (two breeds × two diets). Diets were isonitrogenous and isoenergetic (200 ± 1 g CP/kg dry matter (DM); 14.7 ± 0.1 MJ ME/kg DM) and with identical chemical composition except for lysine concentration (10.9 and 5.20 g lysine/kg DM, for lysine-adequate (AL) diet and lysine-deficient (DL) diet respectively). Pigs were individually housed, and daily feed allowance was adjusted on a weekly basis according to BW. Pigs were slaughtered at 25 kg BW. Isoleucine, valine and phenylalanine concentration were higher in carcass protein of Iberian pigs (p < .01). In longissimus muscle, higher concentration of arginine, isoleucine, phenylalanine, lysine and valine (p < .001-p < .05), and lower of methionine (p < .001) were detected in Iberian pigs, whereas phenylalanine, leucine, lysine, threonine and methionine concentration decreased and arginine increased (p < .001-p < .05) when pigs were fed DL diet. Genotype and lysine deficiency effects were moderate in the AA composition of protein of biceps femoris muscle. The results show that AA proportions in protein of carcass and longissimus muscle can be influenced by pig genotype and conditions of lysine shortage. The biceps femoris muscle, with different functional and metabolic properties, shows more constant AA composition than longissimus, which seem to prevail independent from genotype or nutritional challenges.

Meat quality traits in longissimus lumborum and gluteus medius muscles from immunocastrated and surgically castrated Iberian pigs.

This study analyzes quality traits of longissimus lumborum and gluteus medius in surgically castrated (SC) males, inmunocastrated (IC) males and IC female Iberian pigs reared in intensive conditions (n = 18/sex). Vaccination with Improvac® was applied at 18 and 26 weeks of age and slaughtering at 31 weeks. At such conditions, notable sex differences were found. Both muscles of IC males were less red (P < .01), longissimus had lower intramuscular fat (P < .01) and gluteus less Fe content (P < .01) than SC males and IC females. Loins from IC males also exhibited higher drip losses (2.7% at 24 h) than SC males (1.9%) and IC females (1.6%) (P < .001). Fatty acid composition of muscles differed among sexes, especially polyunsaturated fatty acids, which varied as follows: IC males > IC females > SC males in both muscles. It was concluded that meat quality traits differed between inmunocastrated and surgically castrated male Iberian pigs.

Experiencia familiar de la muerte materna / Family experience of maternal death / Experiência dos parentes diante da morte materna

Objetivo: Conocer y analizar la muerte materna, a partir de una aproximación a la realidad familiar, al indagar la experiencia vivida por los familiares de la mujer fallecida y valorar los cambios en los roles y la dinámica familiar. Materiales y Métodos: La investigación se realizó mediante metodología cualitativa, a través del estudio de casos; éstos fueron diez familias que tuvieron una muerte materna en los últimos cinco años, atendidas en hospitales y clínicas del departamento del Tolima-Colombia. Resultados: Ante una muerte materna, las familias y cada uno de sus miembros se tornan vulnerables. Cuando se cuenta con el apoyo de otros familiares, amigos o vecinos, el jefe de familia tiene un trabajo, o la familia pertenece a un grupo religioso o red de apoyo social, la situación se hace menos traumática. Las relaciones familiares en algunos casos se fortalecen, en otros empeoran, particularmente entre suegras y yernos. Con respecto a los cuidados maternales, normalmente las abuelas son las que continúan desempeñando el rol de cuidadoras o madres sustitutas. Conclusiones: Este tipo de muertes, pueden generar sentimientos contradictorios, por un lado se vive el dolor por la ausencia del familiar fallecido y por otro, la alegría de un nuevo miembro. La familia vive una situación de duelo ante la muerte de alguien que trae y cuida la vida, considerada como una muerte trágica, prematura y evitable, por lo que la espera de recibir a un nuevo miembro es sustituida por una pérdida.

Objective: Understanding and analyzing maternal death, approaching to the family reality by examining the experience of the family members of the deceased, and evaluating the change of roles and family dynamic. Materials and Methods: The research was made through study cases, using qualitative methodology; these were ten families that experienced maternal death in the last 5 years, and were attended in clinics and hospitals in de department of Tolima-Colombia. Results: Facing maternal death, families and each of their members become vulnerable. Having support from other family members, friends or neighbors, having a job, or belonging to a religious group or social support network, makes the situation less traumatic. Family relations in some cases are strengthened or worsened, particularly between the mother-in-law and son-in-law. Regarding maternal care, grandmothers normally take the role of caretakers or substitute mothers. Conclusions: These types of deaths cause contradictory feelings, on one hand there is the pain for the absence of a dead family member and on the other, the joy of a new one. The family goes through a situation of facing the death of someone who brings life and cares for it. It is considered a tragic, premature, and avoidable death, consequently the longing of receiving a new family member is substituted by a loss.

Objetivo: Conhecer e analisar a morte materna, a partir de uma aproximação à realidade da família, ao indagar a experiencia vivida pelos parentes da mulher falecida e valorar as mudanças nos papéis e na dinâmica da família. Materiais e Métodos: A pesquisa se realizou utilizando a metodologia qualitativa, através do estudo de casos; estes foram dez famílias que tiveram uma morte materna nos últimos cinco anos, atendidas em hospitais e clínicas do estado de Tolima, na Colômbia. Resultados: Diante uma morte materna, as famílias e cada um de seus membros se tornam vulneráveis. Quando se conta com o apoio de outros parentes, amigos ou vizinhos, o chefe de família tem um trabalho, ou a família pertence a um grupo religioso ou rede de apoio social, a situação se faz menos traumática. Os relacionamentos da família em alguns casos se fortalecem, em outros pioram, particularmente entre sogras e genros. Com respeito aos cuidados maternais, normalmente as avós são as que continuam desempenhando o papel de cuidadoras ou mães substitutas. Conclusões: Este tipo de mortes, podem gerar sentimentos contraditórios, por um lado se vive a dor pela ausência do parente falecido e por outro, a alegria de um novo membro da família. A família vive uma situação de luto diante da morte de alguém que traz e cuida da vida, considerada como uma morte trágica, prematura e evitável, pelo que a espera de receber a um novo membro na família é substituída por uma perda.

DSM265 for Plasmodium falciparum chemoprophylaxis: a randomised, double blinded, phase 1 trial with controlled human malaria infection.

BACKGROUND: A drug for causal (ie, pre-erythrocytic) prophylaxis of Plasmodium falciparum malaria with prolonged activity would substantially advance malaria control. DSM265 is an experimental antimalarial that selectively inhibits the parasite dihydroorotate dehydrogenase. DSM265 shows in vitro activity against liver and blood stages of P falciparum. We assessed the prophylactic activity of DSM265 against controlled human malaria infection (CHMI). METHODS: At the Institute of Tropical Medicine, Eberhard Karls University (Tübingen, Germany), healthy, malaria-naive adults were allocated to receive 400 mg DSM265 or placebo either 1 day (cohort 1A) or 7 days (cohort 2) before CHMI by direct venous inoculation (DVI) of 3200 aseptic, purified, cryopreserved P falciparum sporozoites (PfSPZ Challenge; Sanaria Inc, Rockville, MD, USA). An additional group received daily atovaquone-proguanil (250-100 mg) for 9 days, starting 1 day before CHMI (cohort 1B). Allocation to DSM265, atovaquone-proguanil, or placebo was randomised by an interactive web response system. Allocation to cohort 1A and 1B was open-label, within cohorts 1A and 2, allocation to DSM265 and placebo was double-blinded. All treatments were given orally. Volunteers were treated with an antimalarial on day 28, or when parasitaemic, as detected by thick blood smear (TBS) microscopy. The primary efficacy endpoint was time-to-parasitaemia, assessed by TBS. All participants receiving at least one dose of chemoprophylaxis or placebo were considered for safety, those receiving PfSPZ Challenge for efficacy analyses. Log-rank test was used to compare time-to-parasitemia between interventions. The trial was registered with ClinicalTrials.gov, number NCT02450578. FINDINGS: 22 participants were enrolled between Oct 23, 2015, and Jan 18, 2016. Five participants received 400 mg DSM265 and two participants received placebo 1 day before CHMI (cohort 1A), six participants received daily atovaquone-proguanil 1 day before CHMI (cohort 1B), and six participants received 400 mg DSM265 and two participants received placebo 7 days before CHMI (cohort 2). Five of five participants receiving DSM265 1 day before CHMI and six of six in the atovaquone-proguanil cohort were protected, whereas placebo recipients (two of two) developed malaria on days 11 and 14. When given 7 days before CHMI, three of six volunteers receiving DSM265 became TBS positive on days 11, 13, and 24. The remaining three DSM265-treated, TBS-negative participants of cohort 2 developed transient submicroscopic parasitaemia. Both participants receiving placebo 7 days before CHMI became TBS positive on day 11. The only possible DSM265-related adverse event was a moderate transient elevation in serum bilirubin in one participant. INTERPRETATION: A single dose of 400 mg DSM265 was well tolerated and had causal prophylactic activity when given 1 day before CHMI. Future trials are needed to investigate further the use of DSM265 for the prophylaxis of malaria. FUNDING: Global Health Innovative Technology Fund, Wellcome Trust, Bill & Melinda Gates Foundation through Medicines for Malaria Venture, and the German Center for Infection Research.

A reassessment of the transition-metal free suzuki-type coupling methodology.

We present here a reassessment of our transition-metal free Suzuki-type coupling protocol. We believe that, although the reaction can be run without the need for addition of a metal catalyst, palladium contaminants down to a level of 50 ppb found in commercially available sodium carbonate are responsible for the generation of the biaryl rather than, as previously suggested, an alternative non-palladium-mediated pathway. We present a revised methodology for Suzuki couplings using ultralow palladium concentrations for use with aryl and vinyl boronic acids and discuss the effects of the purity of the boronic acid on the reaction.

Mutations in CFTR gene and clinical correlation in Argentine patients with congenital bilateral absence of the vas deferens.

Congenital bilateral absence of the vas deferens (CBAVD) is a form of male infertility in which mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene have been identified. Here we identify different mutations of CFTR and the poly-T variant of intron 8 (IVS8) in Argentine patients and analyze sweat test values and clinical characteristic related to Cystic Fibrosis (CF). For counseling purposes the two most frequent mutations in Argentine CF population: deltaF508 and G542X were screened in wives. In all cases, it was possible to reduce the risk of CF/CBAVD descendants in these couples because none of the mutation were found in the 36 samples. Eight patients (23%) showed abnormal chloride values (> 60 mmol/l). A second group of 6 patients (18%) had borderline values of sweat chloride (40-59 mmol/l). We defined another group with 6 patients (18%), with normal sweat chloride levels (30-39 mmo/l) and a fourth group of 14 (41%) patients with sweat chloride below 30 mmol/l. deltaF508, the most frequent CF mutation in the Argentine population, was found on 15 of the 72 chromosomes (21%), R117H mutation was detected on 2 of 62 chromosomes (3%). Only one R347P allele was found on 28 chromosomes analyzed (2%). On a sample of 27 patients, IVS8 analysis showed a frequency of 6/56 chromosomes (11%) of 5T allele. Even though these findings present an improvement in the detection of mutations related to clinical correlations in Argentine CBAVD population, the search for other common and uncommon mutations should be continued.

Mutations in CFTR gene and clinical correlation in argentine patients with congenital bilateral absence of the vas deferens

Correlación de las características clínicas con mutaciones del gen CFTR en pacientes argentinos con ausencia bilateral congénita de vasos deferentes. La ausencia bilateral congénita de vasos deferentes (CBAVD) es una forma de infertilidad masculina en la que se han identificado mutaciones en el gen de la conductancia transmembrana de la fibrosis quística (CFTR). Hemos estudiado en pacientes argentinos diferentes mutaciones en el CFTR y la variante poli T del intron 8 (IVS8) y analizado los valores de test del sudor y las características clínicas relacionadas a la Fibrosis Quística (FQ). Para el asesoramiento genético se han estudiado en las esposas de estos pacientes, las dos mutaciones más frecuentes en la población FQ del país, ∆F508 y G542X. Como no se encontraron mutaciones, el riesgo de descendencia CF/CBAVD fue reducido del 2 al 0.7%. Ocho pacientes (23%) presentaban test del sudor anormales (> 60 mmol/l). Un segundo grupo de 6 pacientes (18%) presentaron valores dudosos (40-59 mmol/l). Hemos definido un tercer grupo de 6 pacientes con valores normales de test del sudor (18%), comprendidos entre los 30 y 39 mmo/l, y un cuarto grupo de 14 pacientes (41%) con valores de cloruro en sudor inferiores a 30 mmol/l. La mutación más frecuente en la población CF argentina, ∆F508, fue encontrada en 15 de los 72 cromosomas (21%) analizados, la R117H fue encontrada en 2 de los 62 cromosomas estudiados (3%). Un único alelo R347P fue encontrado en los 28 cromosomas analizados (2%). De los 27 pacientes a los que se les estudió el tracto IVS8, 6/56 cromosomas (11%) presentaban el alelo 5T. Si bien estos hallazgos representan un avance en relación a la detección de mutaciones correlacionadas con los síntomas clínicos en la población CBAVD argentina, se debe continuar la búsqueda de otras mutaciones comunes y raras con el fin de establecer una conducta terapéutica en estos pacientes.

Mutations in CFTR gene and clinical correlation in Argentine patients with congenital bilateral absence of the vas deferens.

Congenital bilateral absence of the vas deferens (CBAVD) is a form of male infertility in which mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene have been identified. Here we identify different mutations of CFTR and the poly-T variant of intron 8 (IVS8) in Argentine patients and analyze sweat test values and clinical characteristic related to Cystic Fibrosis (CF). For counseling purposes the two most frequent mutations in Argentine CF population: deltaF508 and G542X were screened in wives. In all cases, it was possible to reduce the risk of CF/CBAVD descendants in these couples because none of the mutation were found in the 36 samples. Eight patients (23

) showed abnormal chloride values (> 60 mmol/l). A second group of 6 patients (18

) had borderline values of sweat chloride (40-59 mmol/l). We defined another group with 6 patients (18

), with normal sweat chloride levels (30-39 mmo/l) and a fourth group of 14 (41

) patients with sweat chloride below 30 mmol/l. deltaF508, the most frequent CF mutation in the Argentine population, was found on 15 of the 72 chromosomes (21

), R117H mutation was detected on 2 of 62 chromosomes (3

). Only one R347P allele was found on 28 chromosomes analyzed (2

). On a sample of 27 patients, IVS8 analysis showed a frequency of 6/56 chromosomes (11

) of 5T allele. Even though these findings present an improvement in the detection of mutations related to clinical correlations in Argentine CBAVD population, the search for other common and uncommon mutations should be continued.

Mutations in CFTR gene and clinical correlation in argentine patients with congenital bilateral absence of the vas deferens

Correlación de las características clínicas con mutaciones del gen CFTR en pacientes argentinos con ausencia bilateral congénita de vasos deferentes. La ausencia bilateral congénita de vasos deferentes (CBAVD) es una forma de infertilidad masculina en la que se han identificado mutaciones en el gen de la conductancia transmembrana de la fibrosis quística (CFTR). Hemos estudiado en pacientes argentinos diferentes mutaciones en el CFTR y la variante poli T del intron 8 (IVS8) y analizado los valores de test del sudor y las características clínicas relacionadas a la Fibrosis Quística (FQ). Para el asesoramiento genético se han estudiado en las esposas de estos pacientes, las dos mutaciones más frecuentes en la población FQ del país, ∆F508 y G542X. Como no se encontraron mutaciones, el riesgo de descendencia CF/CBAVD fue reducido del 2 al 0.7%. Ocho pacientes (23%) presentaban test del sudor anormales (> 60 mmol/l). Un segundo grupo de 6 pacientes (18%) presentaron valores dudosos (40-59 mmol/l). Hemos definido un tercer grupo de 6 pacientes con valores normales de test del sudor (18%), comprendidos entre los 30 y 39 mmo/l, y un cuarto grupo de 14 pacientes (41%) con valores de cloruro en sudor inferiores a 30 mmol/l. La mutación más frecuente en la población CF argentina, ∆F508, fue encontrada en 15 de los 72 cromosomas (21%) analizados, la R117H fue encontrada en 2 de los 62 cromosomas estudiados (3%). Un único alelo R347P fue encontrado en los 28 cromosomas analizados (2%). De los 27 pacientes a los que se les estudió el tracto IVS8, 6/56 cromosomas (11%) presentaban el alelo 5T. Si bien estos hallazgos representan un avance en relación a la detección de mutaciones correlacionadas con los síntomas clínicos en la población CBAVD argentina, se debe continuar la búsqueda de otras mutaciones comunes y raras con el fin de establecer una conducta terapéutica en estos pacientes.(AU)