Human pluripotent stem cells (PSCs) provide a promising resource to produce immune cells for adoptive cellular immunotherapy to better treat and potentially cure otherwise lethal cancers. Cytotoxic T ...cells and natural killer (NK) cells can now be routinely produced from human PSCs. These PSC‐derived lymphocytes have phenotype and function similar to primary lymphocytes isolated from peripheral blood. PSC‐derived T and NK cells have advantages compared with primary immune cells, as they can be precisely engineered to introduce improved anti‐tumor activity and produced in essentially unlimited numbers. Stem Cells 2018;36:134–145
Human pluripotent stem cells can be differentiated into lymphocytes with potent anti‐tumor activity. The hESC/iPSC‐derived lymphocytes can be produced at clinical scale and be engineered to provide improved targeting and increased efficiency in killing diverse tumor targets. This strategy allows hESC/iPSC‐derived lymphocytes to provide off‐the‐shelf, targeted effector cells suitable to treat large numbers of cancer patients with standardized immune cell therapy.
Objective
The objective was to evaluate available characteristics and financial costs of malpractice cases among advanced practice providers (APPs; nurse practitioners NPs and physician assistants ...PAs), trainees (medical students, residents, fellows), and attending physicians.
Methods
This study was a retrospective analysis of claims occurring in the emergency department (ED) from January 1, 2010, to December 31, 2019, contained in the Candello database. Cases were classified according to the provider type(s) involved: NP, PA, trainee, or cases that did not identify an extender as being substantially involved in the adverse event that resulted in the case (“no extender”).
Results
There were 5854 cases identified with a total gross indemnity paid of $1,007,879,346. Of these cases, 193 (3.3%) involved an NP, 513 (8.8%) involved a PA, 535 (9.1%) involved a trainee, and 4568 (78.0%) were no extender. Cases where a trainee was involved account for the highest average gross indemnity paid whereas no‐extender cases are the lowest. NP and PA cases differed by contributing factors compared to no‐extender cases: clinical judgment (NP 89.1% vs. no extender 76.8%, p < 0.0001; PA 84.6% vs. no extender, p < 0.0001), documentation (NP 23.3% vs. no extender 17.8%, p = 0.0489; PA 25.9% vs. no extender, p < 0.0001), and supervision (NP 22.3% vs. no extender 1.8%, p < 0.0001; PA 25.7% vs. no extender p < 0.0001). Cases involving NPs and PAs had a lower percentage of high‐severity cases such as loss of limb or death (NP 45.6% vs. no extender 50.2%, p = 0.0004; PA 48.3% vs. no extender, p < 0.0001).
Conclusions
APPs and trainees comprise approximately 21% of malpractice cases and 33% of total gross indemnity paid in this large national ED data set. Understanding differences in characteristics of malpractice claims that occur in emergency care settings can be used to help to mitigate provider risk.
Background
Volume overload is frequently encountered and is associated with cardiovascular risk factors in patients with chronic kidney disease (CKD). However, the relationship between volume ...overload and adverse outcomes in CKD is not fully understood.
Methods and Results
A prospective cohort of 338 patients with stage 3 to 5 CKD was followed for a median of 2.1 years. The study participants were stratified by the presence or absence of volume overload, defined as an overhydration index assessed by bioimpedance spectroscopy exceeding 7%, the 90th percentile for the healthy population. The primary outcome was the composite of estimated glomerular filtration rate decline ≥50% or end‐stage renal disease. The secondary outcome included a composite of morbidity and mortality from cardiovascular causes. Animal models were used to simulate fluid retention observed in human CKD. We found that patients with volume overload were at a higher risk of the primary and secondary end points in the adjusted Cox models. Furthermore, overhydration appears to be more important than hypertension in predicting an elevated risk. In rats subjected to unilateral nephrectomy and a high‐salt diet, the extracellular water significantly increased. This fluid retention was associated with an increase in blood pressure, proteinuria, renal inflammation with macrophage infiltration and tumor necrosis factor‐α overexpression, glomerular sclerosis, and cardiac fibrosis. Diuretic treatment with indapamide attenuated these changes, suggesting that fluid retention might play a role in the development of adverse outcomes.
Conclusions
Volume overload contributes to CKD progression and cardiovascular diseases. Further research is warranted to clarify whether the correction of volume overload would improve outcomes for CKD patients.
We report the derivation of induced pluripotent stem (iPS) cells from adult skin fibroblasts using a single, polycistronic lentiviral vector encoding the reprogramming factors Oct4, Sox2, and Klf4. ...Porcine teschovirus‐1 2A sequences that trigger ribosome skipping were inserted between human cDNAs for these factors, and the polycistron was subcloned downstream of the elongation factor 1 alpha promoter in a self‐inactivating (SIN) lentiviral vector containing a loxP site in the truncated 3′ long terminal repeat (LTR). Adult skin fibroblasts from a humanized mouse model of sickle cell disease were transduced with this single lentiviral vector, and iPS cell colonies were picked within 30 days. These cells expressed endogenous Oct4, Sox2, Nanog, alkaline phosphatase, stage‐specific embryonic antigen‐1, and other markers of pluripotency. The iPS cells produced teratomas containing tissue derived from all three germ layers after injection into immunocompromised mice and formed high‐level chimeras after injection into murine blastocysts. iPS cell lines with as few as three lentiviral insertions were obtained. Expression of Cre recombinase in these iPS cells resulted in deletion of the lentiviral vector, and sequencing of insertion sites demonstrated that remnant 291‐bp SIN LTRs containing a single loxP site did not interrupt coding sequences, promoters, or known regulatory elements. These results suggest that a single, polycistronic “hit and run” vector can safely and effectively reprogram adult dermal fibroblasts into iPS cells. Stem Cells 2009;27:1042–1049
Mutations of the Janus family kinase JAK3 gene cause severe combined immunodeficiency (SCID). JAK3 deficiency in humans is characterized by the absence of circulating T cells and natural killer (NK) ...cells with normal numbers of poorly functioning B cells (T–B+NK–). Using SCID patient-specific induced pluripotent stem cells (iPSCs) and a T cell in vitro differentiation system, we demonstrate a complete block in early T cell development of JAK3-deficient cells. Correction of the JAK3 mutation by CRISPR/Cas9-enhanced gene targeting restores normal T cell development, including the production of mature T cell populations with a broad T cell receptor (TCR) repertoire. Whole-genome sequencing of corrected cells demonstrates no CRISPR/Cas9 off-target modifications. These studies describe an approach for the study of human lymphopoiesis and provide a foundation for gene correction therapy in humans with immunodeficiencies.
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•iPSC modeling of SCID•CRISPR/Cas-enhanced gene correction•Broad T cell receptor repertoire•No off-target modifications
Using SCID patient-specific induced pluripotent stem cells (iPSCs) and a T cell in vitro differentiation system, Chang et al. define a block in early T cell development of JAK3-deficient cells. Correction of the JAK3 mutation by CRISPR/Cas9-enhanced gene targeting restores normal T cell development.
Human induced pluripotent stem cells (hiPSCs) have enormous potential for the treatment of inherited and acquired disorders. Recently, antigen-specific T lymphocytes derived from hiPSCs have been ...reported. However, T lymphocyte populations with broad T cell receptor (TCR) diversity have not been generated. We report that hiPSCs derived from skin biopsy are capable of producing T lymphocyte populations with a broad TCR repertoire. In vitro T cell differentiation follows a similar developmental program as observed in vivo, indicated by sequential expression of CD7, intracellular CD3 and surface CD3. The γδ TCR locus is rearranged first and is followed by rearrangement of the αβ locus. Both γδ and αβ T cells display a diverse TCR repertoire. Upon activation, the cells express CD25, CD69, cytokines (TNF-α, IFN-γ, IL-2) and cytolytic proteins (Perforin and Granzyme-B). These results suggest that most, if not all, mechanisms required to generate functional T cells with a broad TCR repertoire are intact in our in vitro differentiation protocol. These data provide a foundation for production of patient-specific T cells for the treatment of acquired or inherited immune disorders and for cancer immunotherapy.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
IntroductionThe point-of-care lung ultrasound (LUS) score has been used in coronavirus 2019 (COVID-19) patients for diagnosis and risk stratification, due to excellent sensitivity and infection ...control concerns. We studied the ratio of partial pressure of oxygen in arterial blood to the fraction of inspiratory oxygen concentration (PO2/FiO2), intubation rates, and mortality correlation to the LUS score.MethodsWe conducted a systematic review using PRISMA guidelines. Included were articles published from December 1, 2019-November 30, 2021 using LUS in adult COVID-19 patients in the intensive care unit or the emergency department. Excluded were studies on animals and on pediatric and pregnant patients. We assessed bias using QUADAS-2. Outcomes were LUS score and correlation to PO2/FiO2, intubation, and mortality rates. Random effects model pooled the meta-analysis results.ResultsWe reviewed 27 of 5,267 studies identified. Of the 27 studies, seven were included in the intubation outcome, six in the correlation to PO2/FiO2 outcome, and six in the mortality outcome. Heterogeneity was found in ultrasound protocols and outcomes. In the pooled results of 267 patients, LUS score was found to have a strong negative correlation to PO2/FiO2 with a correlation coefficient of -0.69 (95% confidence interval CI -0.75, -0.62). In pooled results, 273 intubated patients had a mean LUS score that was 6.95 points higher (95% CI 4.58-9.31) than that of 379 non-intubated patients. In the mortality outcome, 385 survivors had a mean LUS score that was 4.61 points lower (95% CI 3.64-5.58) than that of 181 non-survivors. There was significant heterogeneity between the studies as measured by the I2 and Cochran Q test.ConclusionA higher LUS score was strongly correlated with a decreasing PO2/FiO2 in COVID-19 pneumonia patients. The LUS score was significantly higher in intubated vs non-intubated patients with COVID-19. The LUS score was significantly lower in critically ill patients with COVID-19 pneumonia that survive.
To gain insight into mechanisms controlling SRY (sex determining region Y)-box 2 (Sox2) protein activity in mouse embryonic stem cells (ESCs), the endogenous Sox2 gene was tagged with ...FLAG/Hemagglutinin (HA) sequences by homologous recombination. Sox2 protein complexes were purified from Sox2/FLAG/HA knockin ESCs, and interacting proteins were defined by mass spectrometry. One protein in the complex was poly ADP-ribose polymerase I (Parp1). The results presented below demonstrate that Parp1 regulates Sox2 protein activity. In response to fibroblast growth factor (FGF)/extracellular signal-regulated kinase (ERK) signaling, Parp1 auto-poly ADP-ribosylation enhances Sox2-Parp1 interactions, and this complex inhibits Sox2 binding to octamer-binding transcription factor 4 (Oct4)/Sox2 enhancers. Based on these results, we propose a unique mechanism in which FGF signaling fine-tunes Sox2 activity through posttranslational modification of a critical interacting protein, Parp1, and balances the maintenance of ESC pluripotency and differentiation. In addition, we demonstrate that regulation of Sox2 activity by Parp1 is critical for efficient generation of induced pluripotent stem cells.
In this work, we established the relationship between the thermally induced microstructure development and the photoluminescence efficiency (E p) of the poly(9,9-dioctylfluorene) (PF8) fibers ...prepared from the semidilute PF8/chloroform solutions by direct electrospinning. The as-spun (AS) fibers were found to be composed of an amorphous phase and a significant fraction of mesomorphic β-phase, which may promote the E p of PF8. The temperature-dependent wide-angle X-ray scattering (WAXS) measurement revealed that the PF8 fibers exhibited complex structural transformation on heating from the AS state, and there existed a strong correlation between E p and the microstructure developed. The AS fibers exhibited the highest E p value. The E p decreased progressively on heating at temperatures below the cold crystallization temperature (T cc), where the microstructure remained unperturbed. The β-phase transformed to γ- or α′-phase (C γ-/C α′-crystals) above T cc, and such a structural transformation led to a reduction of E p. With further heating, the C γ-/C α′-crystals transformed into the more stable C α-crystals, presumably as the premelting–reorganization–remelting proceeded at the temperatures over the crystal transformation temperature (T ct). The crystal transformation process reduced the E p markedly. Finally, above the melting temperature (T m), the C α-crystals were disrupted into the disordered molten state, and the E p value remained largely unchanged up to 200 °C.
We identified the conformational structure, aggregation behavior and photophysical properties of poly2-methoxy-5-(2-ethylhexyloxy)-1,4-phenylenevinylene (MEH-PPV) in various toluene/nonane solutions ...dependent on nonane content. In dilute solution region (e.g., below the overlap concentration, c*), static light scattering (SLS) revealed that the stiffness of MEH-PPV decreased as nonane content was increased, attributed to a stronger interaction force between MEH-PPV backbones. However, MEH-PPV chains presented a more extended-wormlike conformational structure in toluene. Therefore, a looser intermolecular micellelike aggregates occurred easily between the ether side chains of MEH-PPV at the concentration exceeded the overlapping concentration in toluene and toluene/nonane solutions. Nevertheless, at higher nonane content, the micellelike aggregates were became highly compacted by the π–π complex force between MEH-PPV backbones, dominating the dynamic behaviors and photophysical properties of MEH-PPV in toluene/nonane solution. These results proved that two types of segmental association with distinct stability were identified for the MEH-PPV/toluene/nonane solutions upon prolonged aging. The highly stable, which could not be disrupted by heating, was attributed to the interaction force between ether side chains of MEH-PPV in both solutions. Other type was a weaker affinity force through π–π complex, which could be disrupted thermally, making the micellelike aggregates more compact to form the network structure in MEH-PPV-enriched at higher nonane content.
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