Atraumatic Restorative Treatment (ART): a Three-year Community Field Trial in Thailand—Survival of One-surface Restorations in the Permanent Dentition

This study compares the Atraumatic Restorative Treatment (ART) technique to conventional amalgam restorations in the management of dental caries. The present report is limited to the results for one-surface restorations in the permanent dentition over a three-year period. A community field trial was carried out in rural villages in northeastern Thailand. Dental caries was treated using the ART technique in one village where 144 persons were treated with 241 restorations. In a second village, 205 conventional amalgam restorations were provided to 138 persons using mobile dental equipment. Both ART and amalgam restorations were performed by one dentist and two dental nurses without administering local anesthesia. Clinical evaluation was carried out one, two, and three years after placement. The longevity of the restorations was determined by computing the estimated cumulative survival rates according to the life table method. The survival rates of ART restorations (93%, 83%, 71% in years one, two, and three, respectively) were close to those for amalgam restorations (98%, 94%, 85%); however, differences were statistically significant. No statistically significant differences were observed between ART restorations in children and adults, or between those placed by the dentist and dental nurses. Survival rates were lower for occlusal surface restorations compared to those in other surfaces. ART is a feasible approach for the management of dental caries, especially for one-surface lesions in the permanent dentition. Because of its simplicity as a minimal intervention technique, ART can make the control of dental caries available to all people irrespective of their economic and living conditions.     

Expansion of experimental genetics approaches for Plasmodium berghei with versatile transfection vectors

Experimental reverse genetic approaches have proven powerful in the study of the biology of the malaria parasite. The murine malaria model parasite Plasmodium berghei is the genetically most amendable Plasmodium species and allows full access to the entire life cycle in vivo. Here, we describe a next-generation, highly versatile transfection vector set that facilitates advancing experimental genetic strategies towards a genome-wide scale. Through 36 consecutive cloning and 17 subcloning steps an optimized vector set was generated from the standard transfection plasmid. These targeting vectors, collectively referred to as the Berghei Adaptable Transfection (pBAT) plasmids, contain key elements that permit recycling of the drug-selectable cassette, robust green fluorescent labelling of recombinant parasites, carboxy-terminal tagging of target proteins with a red fluorescent-epitope tag fusion, and expression of heterologous genes. The vectors were further optimized for small size, versatile restriction endonuclease recognition sites and potential exchange of individual vector elements. We show that stable integration into a transgene expression site, an intergenic locus at a synteny breakpoint on P. berghei chromosome 6, is phenotypically silent and generated a bright green fluorescent parasite line for imaging applications. We provide an example, P. berghei actin 2, for targeted gene deletion and illustrate that the positive selection marker can be recycled, thereby permitting multiple rounds of genetic manipulations. We propose that the vectors described herein will greatly facilitate functional assignment to predicted and orphan Plasmodium gene models by multiple experimental genetics approaches.     

Early determinants of long-term T-cell reconstitution after hematopoietic stem cell transplantation for severe combined immunodeficiency

The immune system of patients with severe combined immunodeficiency (SCID) reconstitutes to a large extent during the first years after hematopoietic stem cell transplantation (HSCT). It was suggested, however, that accelerated loss of thymus output may cause impaired immune function at the long term. To address this issue, we studied patients with SCID who underwent allogeneic HSCT 5 to 32 years earlier and identified early determinants of long-term T-cell reconstitution. A variety of immune parameters were analyzed both early (1-4 years) and late (5-32 years) after HSCT. Late after HSCT, a clear distinction could be made between a group of 8 patients with impaired T-cell reconstitution and 11 patients with good immune reconstitution. Importantly, in patients with decreased long-term T-cell reconstitution, T-cell recovery was already poor early after HSCT, demonstrating that long-term immune failure was not caused by accelerated loss of thymus output or long-term graft failure, but resulted from poor early grafting. The number of T-cell receptor excision circles (TRECs) early after HSCT was most predictive for long-term T-cell reconstitution. Frequent monitoring of T-cell immunity and TREC numbers early after HSCT may thus serve to timely identify patients who will fail to reconstitute properly and who may need additional treatment.      

Influenza virus vaccination induces interleukin-12/23 receptor beta 1 (IL-12/23R beta 1)-independent production of gamma interferon (IFN-gamma) and humoral immunity in patients with genetic deficiencies in IL-12/23R beta 1 or IFN-gamma receptor I

To investigate whether protective immune responses can be induced in the absence of normal interleukin-12/23/gamma interferon (IL-12/23/IFN-gamma) axis signaling, we vaccinated with the seasonal influenza virus subunit vaccine two patients with complete IL-12/23 receptor beta1 (IL-12/23R beta 1) deficiencies, two patients with partial IFN-gamma receptor I (pIFN-gamma RI) deficiencies, and five healthy controls. Blood samples were analyzed before, 7 days after, and 28 days after vaccination. In most cases, antibody titers reached protective levels. Moreover, although T-cell responses in patients were lower than those observed in controls, significant influenza virus-specific T-cell proliferation, IFN-gamma production, and numbers of IFN-gamma-producing cells were found in all patients 7 days after the vaccination. Interestingly, influenza virus-specific IFN-gamma responses were IL-12/23 independent, in striking contrast to mycobacterium-induced IFN-gamma production. In conclusion, influenza virus vaccination induces IL-12/23-independent IFN-gamma production by T cells and can result in sufficient humoral protection in both IL-12/23R beta 1- and pIFN-gamma RI-deficient individuals.     

Issue Information

Most sickle cell disease (SCD) patients rely on blood transfusion as their main treatment strategy. However, frequent blood transfusion poses the risk of alloimmunization. On average, 30% of SCD patients will alloimmunize while other patient groups form antibodies less frequently. Identification of genetic markers may help to predict which patients are at risk to form alloantibodies. The aim of this study was to evaluate whether genetic variations in the Toll-like receptor pathway or in genes previously associated with antibody-mediated conditions are associated with red blood cell (RBC) alloimmunization in a cohort of SCD patients. In this case-control study, cases had a documented history of alloimmunization while controls had received ≥20 RBC units without alloantibody formation. We used a customized single nucleotide polymorphism (SNP) panel to genotype 690 SNPs in 275 (130 controls, 145 cases) patients. Frequencies were compared using multiple logistic regression analysis. In our primary analysis, no SNPs were found to be significantly associated with alloimmunization after correction for multiple testing. However, in a secondary analysis with a less stringent threshold for significance we found 19 moderately associated SNPs. Among others, SNPs in TLR1/TANK and MALT1 were associated with a higher alloimmunization risk, while SNPs in STAM/IFNAR1 and STAT4 conferred a lower alloimmunization risk.     

Autologous bone versus calcium-phosphate ceramics in treatment of experimental bone defects

Autologous bone grafting is currently considered the treatment of choice for correction of large bone defects. However, to avoid morbidity associated with autologous bone harvesting many artificial bone-substitute materials have been developed over the years. A new generation of resorbable materials is emerging, with promising results so far. In order to investigate the possibility to use one of these new materials as an alternative with better results than hydroxyapatite, an experimental study was performed. A new resorbable calcium phosphate particles and paste forms, the latter of which hardens in situ after application. In 28 sheep, a 3-cm segmental tibial defect was made and intramedullary fixed by an interlocking nail. Twelve weeks after defect filling, radiological, biomechanical, and histological examinations were performed. Mean radiographic and biomechanical tests results were compared with the Mann-Whitney test. Significance was set at p<0.05. Radiographically, the resorbable paste group performed better than all other groups. Biomechanical investigations showed a higher torsional stiffness (p=0.049) for the resorbable calcium-phosphate paste group in comparison with autologous bone. On histological examination, no adverse effects were observed in the calcium-phosphate groups. Resorption by osteoclasts was seen in the resorbable implants. In conclusion, the current study shows an advantageous radiological and mechanical outcome for resorbable calcium phosphates. This indicates that these new materials might be a potential alternative for autologous bone grafting in humans.     

Non-operative management for penetrating splenic trauma: how far can we go to save splenic function?

Background

Selective non-operative management (NOM) for the treatment of blunt splenic trauma is safe. Currently, the feasibility of selective NOM for penetrating splenic injury (PSI) is unclear. Unfortunately, little is known about the success rate of spleen-preserving surgical procedures. The aim of this study was to investigate the outcome of selective NOM for penetrating splenic injuries.

Methods

A dual-centre study is performed in two level-one trauma centres. All identified patients treated for PSI were identified. Patients were grouped based on the treatment they received. Group one consisted of splenectomised patients, the second group included patients treated by a spleen-preserving surgical intervention, and group three included those patients who were treated by NOM.

Results

A total of 118 patients with a median age of 27 and a median ISS of 25 (interquartile range (IQR) 16–34) were included. Ninety-six patients required operative intervention, of whom 45 underwent a total splenectomy and 51 underwent spleen-preserving surgical procedures. Furthermore, 22 patients (12 stab wounds and 10 gunshot wounds) were treated by NOM. There were several anticipated significant differences in the baseline encountered. The median hospitalization time was 8 (5–12) days, with no significant differences between the groups. The splenectomy group had significantly more intensive care unit (ICU) days (2(0–6) vs. 0(0–1)) and ventilation days (1(0–3) vs. 0(0–0)) compared to the NOM group. Mortality was only noted in the splenectomy group.

Conclusions

Spleen-preserving surgical therapy for PSI is a feasible treatment modality and is not associated with increased mortality. Moreover, a select group of patients can be treated without any surgical intervention at all.

     

The influence of hypoxia and fibrinogen variants on the expansion and differentiation of adipose tissue-derived mesenchymal stem cells

Upon implantation of tissue-engineered scaffolds, hypoxia will occur until neovascularization takes place. In vivo, the temporary fibrin matrix forms a suitable matrix for this process and fibrin variants can influence the extent of neovascularization. In this study, the influence of oxygen tension and naturally occurring fibrinogen variants on adipose tissue-derived mesenchymal stem cell (ASC) expansion and differentiation were determined. ASC proliferated 1.7-fold faster in 1% oxygen and showed reduced cell aging, and their stemness was preserved. The stem cell surface marker expression was similar in 1% and 20% oxygen. The various fibrinogen coatings did not influence ASC expansion and differentiation. Differentiation of ASC toward adipogenic and osteogenic lineages was improved in 20% oxygen, whereas 1% oxygen improved chondrogenic differentiation. In conclusion, optimal oxygen concentrations vary for the intended ASC application, and fibrinogen variants, which can be used to influence neovascularization, do not alter ASC behavior. These data emphasize the importance of oxygen concentrations during stem cell growth and differentiation.