Manufacture of Chimeric Antigen Receptor T Cells from Mobilized Cyropreserved Peripheral Blood Stem Cell Units Depends on Monocyte Depletion

Cytotoxic chemotherapy and radiation can render lymphocyte repertoires qualitatively and quantitatively defective. Thus, heavily treated patients are often poor candidates for the manufacture of autologous chimeric antigen receptor (CAR)-T cell products. In the United States and Europe, children with high-risk neuroblastoma undergo apheresis early in the course of treatment to collect peripheral blood stem cells (PBSCs) for cryopreservation in preparation for high-dose chemotherapy followed by autologous stem cell rescue. Here, we investigate whether these cryopreserved chemotherapy and granulocyte colony-stimulating factor (G-CSF)-mobilized PBSCs can serve as starting material for CAR-T cell manufacturing. We evaluated T cell precursor subsets in cryopreserved PBSC units from 8 patients with neuroblastoma using fluorescent activated cell sorting-based analysis. Every cryopreserved unit collected early in treatment contained both CD4 and CD8 precursors with significant numbers of naïve and central memory precursors. Significant numbers of Ki67⁺/PD1⁺ T cells were detected, presumably the result of chemotherapy-induced lymphopenia and subsequent homeostatic proliferation. Cryopreserved PBSC units containing 56 to 112?×?10⁶ T cells were amenable to immunomagnetic selection, CD3?×?28 bead activation, lentiviral transduction, and cytokine-driven expansion, provided that CD14 monocytes were depleted before the initiation of cultures. Second- and third-generation CD171 CAR⁺ CD4 and CD8 effector cells derived from cryopreserved units displayed antineuroblastoma lytic potency and cytokine secretion comparable to those derived from a healthy donor and mediated in vivo antitumor regression in NSG mice. We conclude that cryopreserved PBSCs procured via standard methods during early treatment can serve as an alternative starting source for CAR-T cell manufacturing, extending the options for heavily treated patients.     

High-risk hpv infection in sexually active and non active young women

To determine the prevalence of high-risk HPV infection before first vaginal intercourse and after initiation of sexual activity in 14-22 years old girls, to find which HPV types occur most commonly. During the gynecologic exam cervical swabs were collected. Specimens were tested using Hybrid Capture 2 DNA test. In positive samples digene HPV genotyping test was performed. Subjects included 264 women, of whom 169 (64.0%) reported previous sexual activity and 95 (36.0%) had not yet had vaginal intercourse. There were 43 cases (16.3%) of high risk HPV, with 39 cases in those reporting sexual activity and 4 in virgins. Single strain HPV infection was detected in 62.8%, two strains in 23.2%, and three strains in 14.0% of subjects. HPV-16 was the most common type (20.9%), next more prevalent types were 18, 56, 31, 33 and 59. HPV-16 or HPV-18 were detected in 39.5% of the subjects. Not a single case was identified containing both types. The prevalence of high-risk HPV is common in sexually active young Lithuanian women. HPV were found in a small percentage of girls who had never had vaginal intercourse. The prevalence of types 16 and 18 among Lithuanian women was higher than in international data.     

International variation in the prevalence of preclinical colorectal cancer: Implications for predictive values of noninvasive screening tests and potential target populations for screening

Screening for colorectal cancer (CRC) is implemented in an increasing number of countries. We aimed to assess international variation in the prevalence of preclinical CRC and the resulting variation in positive and negative predictive values (PPVs, NPVs) of existing and potential CRC screening tests in various countries. Using age‐ and sex‐specific CRC incidence data and transition rates from preclinical to clinical CRC we estimated overall and age‐ and sex‐specific prevalence of preclinical CRC in the target population aged 50–74 years in different parts of the world. These prevalence estimates were used to derive PPVs and NPVs for existing and potential noninvasive screening tests with varying levels of sensitivity and specificity. Within all regions and countries, prevalence strongly increases with age and is higher in men than in women. In addition, major variation was seen between regions and countries, with overall prevalence varying between 1 and 0.1%. As a result, PPVs are expected to strongly vary between ～10% for men in high incidence countries, such as Australia and Germany, and 1% for women in low incidence countries, whereas NPVs are expected to be consistently well above 99%. Variation in CRC prevalence profoundly affects expected PPVs of screening tests, and PPVs should be carefully considered when decisions on screening tests and strategies are made for specific populations and health care systems. Here, we provide estimates of preclinical CRC and expected PPVs and NPVs of noninvasive screening tests, which may enhance the empirical basis for planning of population‐based CRC screening strategies.     

Loss-of-function variants of the filaggrin gene are associated with atopic eczema and associated phenotypes in Swedish families

Recent studies have identified 2 loss-of-function variants, R501X and 2282del4, in the filaggrin gene as predisposing factors in the development of eczema. In this study, representing the first analysis of the variants in a Swedish population, we analysed transmission in 406 multiplex eczema families with mainly adult patients. In accordance with previous studies we found association between the filaggrin gene variants and atopic eczema (p=9.5 x 10(-8)). The highest odds ratio for the combined allele, 4.73 (1.98-11.29), p=3.6 x 10(-8), was found for the subgroup with a severe eczema phenotype, and association was also found with raised allergen-specific IgE, allergic asthma and allergic rhinoconjunctivitis occurring in the context of eczema. Our results support an important role for the filaggrin gene variants R501X and 2282del4 in the development and severity of atopic eczema and indicate a possible role for the subsequent progression into eczema-associated phenotypes.     

Designing red-fluorescent superparamagnetic nanoparticles by conjugation with gold clusters

Photoluminescent (PL) metal and metal oxide nanoclusters (NCs), with a size of just several nanometers, are a separate class of nanomaterials abundant with new attractive optical, physical, and chemical properties and biocompatibility. However, the synthesis of PL magnetic NCs via attachment of PL NCs to iron oxide-based nanoparticles (NPs) is still problematic. Motivated by this, herein, we report the development of a microwave-driven conjugation approach of red-fluorescent gold nanoclusters (BSA@AuNCs) to superparamagnetic NPs. Synthesized CoFe₂O₄@AuNCs possess strong photoluminescence in water and ethanol media as well as good colloidal and optical stability, and magnetization response. High-resolution transmission electron microscopy (HRTEM), steady-state and time-resolved photoluminescence spectroscopy, X-ray powder diffraction (XRD), and magnetic measurements from ambient to cryogenic temperatures were applied for structural characterization and evaluation of optical and magnetic properties of the synthesized species.

We report the study of gold clusters'' attachment to magnetic nanoparticles by microwave processing for red-luminescent magnetic species formation.     

Rehabilitating a Loved One: A Personal Story

Fortunately, during their professional lives, only a small percentage of nurses will face the traumatic task of helping a loved one overcome a major physical handicap. In this article, an experienced rehabilitation nurse relates her experience in helping her aged mother cope with the reality of a severe injury and amputation.     

The frequency of PKU and hyperphenylalaninemia in Sweden - a study in institutions for the mentally retarded as well as in neonates

In a country-wide screening examination in Sweden, PKU was observed in 75 (4.5 per 1,000) and hyperphenylalaninemia in 14 (0.8 per 1,000) of a total of 16,805 patients in institutions for the mentally retarded. In the national neonatal screening program in 1965–1975, which covered nearly 1 million individuals, PKU was observed in 36 and hyperphenylalaninemia in 19 individuals. The incidences of PKU and hyperphenylalaninemia in newborns are calculated to be 1/26,000 and 1/50,000, respectively. In addition, PKU and hyperphenylalaninemia were diagnosed in 14 and 1 non-institutionalized individuals, respectively, born before September 1965.
The total numbers of individuals detected as having PKU and hyperphenylalaninemia in Sweden in the present investigation are 125 and 34, respectively.     

Associations of short-term exposure to ambient air pollutants with emergency ambulance calls for the exacerbation of essential arterial hypertension

We investigated the association between daily emergency ambulance calls (EAC) for elevated blood pressure that occurred during the time intervals of 8:00–13:59, 14:00–21:59, and 22:00–7:59, and exposure to CO, PM₁₀, and ozone. We used Poisson regression to explore the association between the risk of EAC and short-term variation of pollutants, adjusting for seasonality and weather variables. Before noon, the risk was associated with an interquartile range (IQR) (7.9 μg/m³) increase in PM₁₀ at lag 2–4 days below the median (RR = 1.08, p = 0.031) and with an IQR (0.146 mg/m³) increase in CO at lag 6–7 below the median (RR = 1.05, p = 0.028). During 14:00–21:59, the risk was associated with an IQR (18.8 μg/m³) increase in PM₁₀ on the previous day below the median (RR = 1.04, p = 0.031). At night, EAC were negatively affected by lower O₃ (lag 0–2) below the median (per IQR decrease RR = 1.10, p = 0.018) and a higher PM₁₀ at lag 0–1 above the median for the elderly (RR = 1.07, p = 0.030).