Preclinical full-scale evaluation of dendritic cells transfected with autologous tumor-mRNA for melanoma vaccination

Most cancer vaccines to date have made use of common tumor antigens or allogenic cancer cell lines. The majority of tumor antigens may, however, be unique patient-specific antigens. Dendritic cells (DCs) are the most potent antigen-presenting cells known. The present report is a full-scale preclinical evaluation of autologous DCs transfected with autologous tumor-mRNA (tDCs) for vaccination in malignant melanoma. By using autologous tumor-mRNA, we intend to make the DCs present a broad spectrum of tumor-associated antigens relevant to each individual patient. Previously, we have described effective methods for mRNA-transfection into DCs by square-wave electroporation and for generating large numbers of DCs. Here, we demonstrate the ability of tDCs, made under full-scale vaccine conditions, to generate in vitro T-cell responses specific for antigens encoded by the transfected tumor-mRNA. T-cell proliferation assays demonstrated tDC-specific responses for all six patients tested. Responses were further studied by IFNgamma ELISPOT and Bioplex cytokine assays (two patients) and by experiments on isolated CD4(+) and CD8(+) T cells, including HLA-blockage (one patient). Moreover, we describe the results of extensive tumor-RNA analysis using Agilent Bioanalyser, a method that we have implemented in the clinical protocol. Based on this preclinical evaluation, a vaccine trial has been started.     

Influence of failure of primary wound healing on subsequent recurrence of pilonidal sinus. combined prospective study and randomised controlled trial.

OBJECTIVE: To find out whether failure of primary wound healing after excision and primary suture for chronic pilonidal sinus predicts recurrence. DESIGN: Follow-up of one prospective study and one subsequent randomised, multicentre study. SETTING: Three teaching hospitals in WesternNorway. SUBJECTS: A total of 197 consecutive patients operated on for chronic pilonidal sinus. INTERVENTIONS: Fifty two patients in the prospective group were given cloxacillin perioperatively. In the randomised study, 145 patients were randomised to have either a single preoperative dose of cefoxitin 2 g intravenously (n = 73) or no prophylaxis (n = 72). Patients were followed up for a median of 7 years. MAIN OUTCOME MEASURES: Recurrence of pilonidal sinus. RESULTS: In the prospective group there were 10 recurrences (19%). In the randomised study 6 patients (8%) who had been given antibiotic prophylaxis had a recurrence compared with 14 patients (19%) who had not been given prophylaxis (p = 0.09). In both groups, failure of primary normal healing was significantly associated with early recurrence (p = 0.0002). Neither the use of antibiotics nor sex had any significant influence on the incidence of recurrences. Most recurrences occurred within the first year. CONCLUSION: Wound complications significantly influenced the recurrence rate whereas antibiotics did not. Most recurrences were seen early.     

Metal-Organic Frameworks (MOFs)-Based Nanomaterials for Drug Delivery

The composition and topology of metal-organic frameworks (MOFs) are exceptionally tailorable; moreover, they are extremely porous and represent an excellent Brunauer–Emmett–Teller (BET) surface area (≈3000–6000 m²·g⁻¹). Nanoscale MOFs (NMOFs), as cargo nanocarriers, have increasingly attracted the attention of scientists and biotechnologists during the past decade, in parallel with the evolution in the use of porous nanomaterials in biomedicine. Compared to other nanoparticle-based delivery systems, such as porous nanosilica, nanomicelles, and dendrimer-encapsulated nanoparticles, NMOFs are more flexible, have a higher biodegradability potential, and can be more easily functionalized to meet the required level of host–guest interactions, while preserving a larger and fully adjustable pore window in most cases. Due to these unique properties, NMOFs have the potential to carry anticancer cargos. In contrast to almost all porous materials, MOFs can be synthesized in diverse morphologies, including spherical, ellipsoidal, cubic, hexagonal, and octahedral, which facilitates the acceptance of various drugs and genes.     

Childhood bullous pemphigoid: a clinicopathologic study and review of the literature

Bullous pemphigoid (BP) is an acquired bullous disorder that predominantly affects the elderly. It is rare in children but when it occurs, there is considerable clinical and histologic overlap with other acquired or congenital blistering disorders. A definitive diagnosis of childhood BP requires direct immunofluorescence and, in some cases, characterization of the target antigen. Three cases of childhood BP are presented, with their histologic and immunofluorescence findings. The first was a 5-month-old male infant who presented with erythema and bullae of the palms and soles and was found to have linear deposition of IgG and C3 along the dermoepidermal junction on direct immunofluorescence (DIF). Histopathologic examination revealed a subepidermal blister containing eosinophils. Type IV collagen was demonstrated along the floor of the blister cavity by a direct immunoperoxidase technique. The second case was an 8-month-old female infant who presented with a blistering eruption of her palms and soles that then became widespread. Direct immunofluorescence showed linear IgG and C3 at the dermoepidermal junction, with laminin deposition at the base of the blister. The third case was a 7-year-old female with bullae and erosions on the vulva and vaginal mucosa. A subepidermal blister was seen on microscopic examination whereas immunofluorescence demonstrated linear IgG and C3 deposition at the basement membrane zone (BMZ). A literature review uncovered 50 cases of childhood BP confirmed by direct or indirect immunofluorescence, or both, and often with evidence of autoantibodies against either the 180 kD or the 230 kD human bullous pemphigoid antigens (BP180 or BP230). This review was used to delineate characteristics of childhood BP, including the newly proposed subtypes: infantile BP and childhood localized vulval BP. Infantile BP presents within the first year of life and is characterized by BP-like lesions on erythematous or normal acral skin. Localized vulval BP is a self-limited, nonscarring BP-like process that involves only the vulva. Both subtypes are normally self-limited and respond well to either topical or systemic steroids, if treatment is initiated before the disease becomes widespread.     

Successful Transplantation of Human Kidneys Deemed Untransplantable but Resuscitated by Ex Vivo Normothermic Machine Perfusion

We report the successful transplantation of a pair of human kidneys that were declined for transplantation due to inadequate in situ perfusion but subsequently transplanted after perfusion and assessment using ex vivo normothermic perfusion (EVNP). The kidneys were from a 35‐year‐old man, a donation after circulatory death donor. Both kidneys were declined by all UK transplant centers. On arrival, the kidneys had significant areas of incomplete clearance of blood from the microcirculation that did not clear after a further attempt to flush them. Kidneys underwent 60 min of EVNP with an oxygenated packed red blood cell–based solution warmed to 35.2°C. During EVNP, the patchy areas cleared in both kidneys. The mean renal blood flow and total urine output were 68.0 mL/min/100 g and 560 mL in the left kidney and 59.9 mL/min/100 g, 430 mL in the right, respectively. Based on the EVNP perfusion parameters, both kidneys were deemed suitable for transplantation. They were transplanted without any complications, and both recipients had initial graft function. The serum creatinine levels at 3 months were 1.2 mg/dl in the recipient of the left kidney and 1.62 mg/dl in the recipient of the right kidney. EVNP technology can be used to assess and rescue kidneys previously deemed unsuitable for transplantation.     

The radioprotective effect of Zataria multiflora against genotoxicity induced by γ irradiation in human blood lymphocytes

The radioprotective effect of hydroalcholic Zataria multiflora (Avishan-e shirazi) extract was investigated against genotoxicity induced by γ irradiation in human lymphocytes. Peripheral blood samples were collected from human volunteers and incubated with Z. multiflora extract at different concentrations (5, 10, and 50?μg/mL) for 1 hour. At each dose point, the whole blood was exposed in vitro to 150 cGy of cobalt-60 γ irradiation, and then the lymphocytes were cultured with mitogenic stimulation to determine number of the micronuclei in cytokinesis-blocked binucleated cells. The treatment of lymphocytes with extract showed a significant decrease in the incidence of micronuclei binucleated cells, compared with similarly irradiated lymphocytes without extract against γ irradiation. The maximum protection and decrease in frequency of micronuclei was observed at 50?μg/mL of Zataria extract by 32% reduction. High-performance liquid chromatography analysis of extract showed that it contains high amounts of thymol. Zataria extract exhibited concentration-dependent radical-scavenging activity on 1,1-diphenyl-2-picryl hydrazyl free radicals. These data have an important application for the protection of human lymphocyte from the genetic damage and side-effects induced by γ irradiation in personnel exposed to radiation.