Transient PU.1 low fetal progenitors generate lymphoid progeny that contribute to adult immunity.

Hematopoietic stem cells and multipotential progenitors emerge in multiple, overlapping waves of fetal development. Some of these populations seed the bone marrow and sustain adult B- and T-cell development long-term after birth. However, others are present transiently, but whether they are vestigial or generate B and T cells that contribute to the adult immune system is not well understood. We now report that transient fetal progenitors distinguished by expression of low levels of the PU.1 transcription factor generated activated and memory T and B cells that colonized and were maintained in secondary lymphoid tissues. These included the small and large intestines, where they may contribute to the maintenance of gut homeostasis through at least middle age. At least some of the activated/memory cells may have been the progeny of B-1 and marginal zone B cells, as transient PU.1low fetal progenitors efficiently generated those populations. Taken together, our data demonstrate the potential of B- and T-cell progeny of transient PU.1low fetal progenitors to make an early and long-term contribution to the adult immune system.

Clinicopathological and Genomic Identification of Breast Cancers with No Impact on Mortality.

BACKGROUND: Implementing mammogram screening means that clinicians are seeing many breast cancers that will never develop metastases. The purpose of this study was to identify subgroups of breast cancer patients who did not present events related to long-term breast cancer mortality, taking into account diagnosis at breast screening, absence of palpability and axillary involvement, and genomic analysis with PAM50. PATIENTS AND METHODS: To identify them, a retrospective observational study was carried out selecting patients without any palpable tumor and without axillary involvement, and a genomic analysis was performed with PAM50. RESULTS: The probability of distant metastasis-free interval (DMFI) of 337 patients was 0.92 (95% CI, 0.90-0.93) at 20 years and 0.96 (95% CI, 0.92-1.00) in 95 patients (28%) with available PAM50 tests. In 22 (23.15%) luminal A tumors and in 9 (9.47%) luminal B tumors smaller than 1 cm, and in HER2 and basal type tumors, there were no metastatic events (20-year DMFI of 1.00). CONCLUSION: Patients with nonpalpable breast cancer found at screening with negative nodes are at very low risk. It is possible to identify subgroups without metastatic events by determining the intrinsic subtype and tumor size less than 1 cm. Therefore, de-escalation of treatment should be considered.

[Association between reproductive history, breast cancer subtype, and survival in premenopausal women]. / Asociación entre la historia reproductiva, el subtipo de cáncer de mama y la supervivencia de mujeres premenopáusicas.

BACKGROUND AND OBJECTIVE: Reproductive history influences breast cancer risk. We analysed its association with tumour subtype and survival in premenopausal women. PATIENTS AND METHODS: Retrospective, observational study of premenopausal women with stage I-III breast carcinoma in the last 20 years. Review of reproductive history, clinical data, and treatments in health records. RESULTS: In 661 premenopausal women (32.40% of 1377 total cases), median age was 47 years (19-53), menarche 12 (7-17), first delivery 28 (16-41) and number of deliveries 2 (0-9). One hundred and eleven (18.20%) were nulliparous. Three hundred and fifty-nine (58.80%) used natural lactation, with a median duration of 6 months. Anovulatory drugs were used by 271 (44.40%), with a median duration of 36 months. Associations were found between menarche <10 years and lower risk of luminal subtype (OR: 0.52, 95% CI: 0.28-0.94; P=.03), between menarche >11 years and lower risk of HER2 subtype (OR: 0.50, 95% CI: 0.26-0.97; P=.04) and between first birth >30 years and lower risk of triple negative subtype (OR: 0.40, 95% CI: 0.17-0.93; P=.03). The 20-year overall and disease-free survival probabilities were 0.80 (95% CI: 0.71-0.90) and 0.72 (95% CI: 0.64-0.79) respectively. Patients with ≥1 delivery had better overall survival than nulliparous patients (HR: 0.51, 95% CI: 0.27-0.96, P=.04). CONCLUSIONS: The findings suggest an association between age at menarche and age at first delivery and breast cancer subtype. Nulliparity is associated with worse survival.

Age-period-cohort analysis of lung cancer mortality inequalities in Southern Spain: missed opportunities for implementing equitable tobacco control policies.

BACKGROUND: Lung cancer mortality in European countries shows different epidemiological patterns according to sex and socioeconomic variables. Some countries show decreasing rates in both sexes, while others show a delayed profile, with increasing mortality in women, inconsistently influenced by socioeconomic status. Our aim was to evaluate the effect of age, period and birth cohort on lung cancer mortality inequalities in men and women in Andalusia, the southernmost region in Spain. METHODS: We used the Longitudinal Database of the Andalusian Population, which collects demographic and mortality data from the 2001 census cohort of more than 7.35 million Andalusians, followed up between 2002 and 2016. Mortality rates were calculated for men and women by educational level, and small-area deprivation. Poisson models were used to assess trends in socioeconomic inequalities in men and women. Finally, age-period-cohort (APC) models were used separately for each educational level and gender. RESULTS: There were 39,408 lung cancer deaths in men and 5,511 in women, yielding crude mortality rates of 78.1 and 11.4 × 105 person-years, respectively. In men higher mortality was found in less educated groups and inequalities increased during the study period: i.e. the rate ratio for primary studies compared to university studies increased from 1.30 (CI95:1.18-1.44) to 1.57 (CI95:1.43-1.73). For women, educational inequalities in favour of the less educated tended to decrease moderately. In APC analysis, a decreasing period effect in men and an increasing one in women were observed. Cohort effect differed significantly by educational level. In men, the lower the educational level, the earlier the peak effect was reached, with a 25-year difference between the least-educated and college-educated. Conversely, college-educated women reached the peak effect with a 12-year earlier cohort than the least-educated women. The decline of mortality followed the same pattern both in men and women, with the best-educated groups experiencing declining rates with earlier birth cohorts. CONCLUSIONS: Our study reveals that APC analysis by education helps to uncover changes in trends occurring in different socioeconomic and gender groups, which, combined with data on smoking prevalence, provide important clues for action. Despite its limitations, this approach to the study of lung cancer inequalities allows for the assessment of gaps in historical and current tobacco policies and the identification of population groups that need to be prioritised for public health interventions.

The layered development of mouse B and T Cells.

Traditional models of lymphopoiesis present B and T cell development as a linear process that initiates in the fetus and continues after birth in the bone marrow and thymus, respectively. However, this view of lymphocyte development is not in accord with reports, dating back several decades, indicating that the types of lymphocytes generated before and after birth differ. In this regard, selected γδ T cells, and those that utilize the Vγ3 receptor in particular, and innate-like B-1 B cells preferentially arise during fetal blood cell development. This review synthesizes data from multiple laboratories, with an emphasis on our own work using mouse models, demonstrating that innate and conventional B and T cells emerge in hematopoietic stem cell independent and dependent waves of development that are differentially regulated. This layering of lymphocyte development has implications for understanding the composition of the adult immune system and may provide insights into the origin of various lymphocytic leukemias.

Clinicopathological characteristics and survival results of patients with ultralow risk breast cancer. / Características clínico-patológicas y supervivencia de pacientes con cáncer de mama de riesgo muy bajo.

BACKGROUND AND OBJECTIVE: To identify subgroups with good progress over an extended period, we used diagnostic screening, tumour palpability, tumour phenotype, and node involvement. PATIENTS AND METHODS: We identified patients with good progress by means of a descriptive, observational and retrospective study. RESULTS: Of 746 patients diagnosed with node-negative breast cancer between 2001 and 2015: 110 (14.75%) had non-palpable screening-diagnosed tumours; 88 (80%) were endocrine-sensitive, 10 (9.10%) were triple-negative and 11 (10%) were HER2. Only 3 patients developed metastases, and there were 4 deaths: 2 from breast cancer and 2 from other causes. The distant recurrence-free interval (DRFI) was 95.60%: 100% in 34 endocrine-sensitive histological grade 1 (equivalent to luminal A) tumours, and 94.40% (95% CI 86.76-102.04) in 54 grade 2-3 (luminal B) tumours. In triple-negative and HER2 cases, it was 100%. In tumours <1 cm it was 100%, and >1 cm it was 95.50% (95% CI 79.42-100.98). CONCLUSIONS: Patients with non-palpable tumours detected by mammogram screening have ultralow risk. The good progress in the luminal A, triple-negative, HER2, and less than 1 cm subgroups may explain the efficacy of the treatment but it also makes them candidates to de-escalation of their treatment.

Targeted and untargeted analysis of triazole fungicides and their metabolites in fruits and vegetables by UHPLC-orbitrap-MS2.

Two extraction methods based on solid liquid extraction and Quick, Easy, Cheap, Effective, Rugged and Safe procedure were developed for the determination of 21 triazole compounds and 5 metabolites, including triazole derivative metabolites as 1,2,4-triazole and 1,2,4-triazol 1-yl-acetic, in courgette, orange, grape and strawberry. The analysis was performed in 10.5 min, using ultra-high performance liquid chromatography coupled to Q-Orbitrap mass analyser. The proposed method was validated according to SANTE 12682/2019. Limits of quantification were ≤10 µg kg-1 for all the compounds, except for 1,2,4-triazol, 1,2,4-triazol 1-yl-acetic, difenoconazole-alcohol and prothioconazole that were 50 µg kg-1. Finally, the method was successfully applied to the analysis of 30 samples. More than 30% of these samples contained residues of triazole compounds. The fungicide most frequently found was myclobutanil. Furthermore, a suspect screening analysis was carried out to search pesticides present in the samples, detecting some of them at concentrations higher than Maximum Residue Limits.

Challenges for hospital management in supporting nurses to deliver humanized care.

Hospitals are paying increasing attention to the delivery of humanized care. The purpose of this study was to explore from the nursing perspective what hospital managers might do to facilitate this. A secondary analysis from a primary ethnographic study regarding dignity in nursing practice was conducted. Twenty interviews of internal medicine nurses from four hospitals were analyzed, and three main themes were identified: Management of nursing teams, Management of ethical values, and Management of the context. It is important for institutional values to be closely aligned with those of the nursing profession, and nurse managers play a key role in ensuring that the latter are applied in practice. The proposed actions offer a cost-effective framework through which nurses and managers may promote the delivery of humanized care.

Nursing students on the frontline: Impact and personal and professional gains of joining the health care workforce during the COVID-19 pandemic in Spain.

BACKGROUND: Final-year nursing students in Spain augmented the health care workforce during the COVID-19 pandemic. PURPOSE: To understand the lived experience of nursing students who joined the health care workforce during the first wave of the COVID-19 outbreak (March-May 2020). METHOD: Qualitative content analysis of the reflective journals of 40 nursing students in Spain. FINDINGS: The analysis identified four main themes: 1) Willingness to help; 2) Safety and protective measures: Impact and challenges; 3) Overwhelming experience: Becoming aware of the magnitude of the epidemic; and 4) Learning and growth. DISCUSSION: The wish to help, the sense of moral duty, and the opportunity to learn buffered the impact of the students' lived experience. Despite the challenges they faced, they saw their experiences as a source of personal and professional growth, and they felt reaffirmed in their choice of career. Promoting opportunities for reflection and implementing adequate support and training strategies is crucial for building a nursing workforce that is capable of responding to future health crises.

UV-A activation of peroxymonosulfate for the removal of micropollutants from secondary treated wastewater.

The occurrence of micropollutants (MPs) in the aquatic environment poses a threat to the environment and to the human health. The application of sulfate radical-based advanced oxidation processes (SR-AOPs) to eliminate these contaminants has attracted attention in recent years. In this work, the simultaneous degradation of 20 multi-class MPs (classified into 5 main categories, namely antibiotics, beta-blockers, other pharmaceuticals, pesticides, and herbicides) was evaluated for the first time in secondary treated wastewater, by activating peroxymonosulfate (PMS) with UV-A radiation, without any pH adjustment or iron addition. The optimal PMS concentration to remove the spiked target MPs (100 µg L-1) from wastewater was 0.1 mM, leading to an average degradation of 80% after 60 min, with most of the elimination occurring during the first 5 min. Synergies between radiation and the oxidant were demonstrated and quantified, with an average extent of synergy of 69.1%. The optimized treatment was then tested using non-spiked wastewater, in which 12 out of the 20 target contaminants were detected. Among these, 7 were degraded at some extent, varying from 10.7% (acetamiprid) to 94.4% (ofloxacin), the lower removals being attributed to the quite inferior ratio of MPs to natural organic matter. Phytotoxicity tests carried out with the wastewater before and after photo-activated PMS oxidation revealed a decrease in the toxicity and that the plants were able to grow in the presence of the treated water. Therefore, despite the low degradation rates obtained for some MPs, the treatment effectively reduces the toxicity of the matrix, making the water safer for reuse.

Use of Busulfan to Condition Mice for Bone Marrow Transplantation.

Myeloablative gamma irradiation has traditionally been used to condition mice prior to bone marrow transplantation. However, irradiation induces high levels of inflammation that may alter patterns of reconstitution. In addition, gamma irradiators are being removed from many facilities for security reasons. Alternative conditioning regimens are thus needed. Here, we describe a protocol for the use of busulfan to condition mice for bone marrow transplantation and several of the variables to consider for effective implementation. For complete details on the use and execution of this protocol, please refer to Montecino-Rodriguez et al. (2019).

Human pediatric B-cell acute lymphoblastic leukemias can be classified as B-1 or B-2-like based on a minimal transcriptional signature.

The finding that transformed mouse B-1 and B-2 progenitors give rise to B-cell acute lymphoblastic leukemias (B-ALLs) with varied aggressiveness suggests that B-cell lineage might also be a factor in the initiation and progression of pediatric B-ALLs in humans. If this is the case, we hypothesized that human pediatric B-ALLs would share gene expression patterns with mouse B-1 or B-2 progenitors. We tested this premise by deriving a distinct 30-gene B-1 and B-2 progenitor signature that was applied to a microarray data set of human pediatric ALLs. Cluster analysis revealed that CRLF2, E2A-PBX1, ERG, and ETV6-RUNX1 leukemias were B-1-like, whereas BCR-ABL1, hyperdiploid, and MLL leukemias were B-2-like. Examination of the 30-gene signature in two independent data sets of pediatric ALLs supported this result. Our data suggest that common genetic subtypes of human ALL have their origin in the B-1 or B-2 lineage.

Proteomics in Deaths by Drowning: Diagnostic Efficacy of Apolipoprotein A1 and α-1Antitrypsin, Pilot Study.

Drowning is one of the leading causes of death worldwide. The pathophysiology of drowning is complex and, sometimes, interpretation of the circumstances of death in the autopsy becomes the main source of information in its diagnosis. New advances in medical research, such as proteomics, especially in forensic pathology, are still in the development. We proposed to investigate the application of Mass Spectrometry-based technologies, to identify differentially expressed proteins that may act as potential biomarkers in the postmortem diagnosis of drowning. We performed a pilot proteomic experiment with the inclusion of two drowned and two control forensic cases. After applying restrictive parameters, we identified apolipoprotein A1 (ApoA1) and α-1 antitrypsin as differentially expressed between the two diagnostic groups. A validation experiment, with the determination of both proteins in 25 forensic cases (16 drowned and 9 controls) was performed, and we corroborated ApoA1 higher values in the drowning group, whereas α-1 antitrypsin showed lower levels. After adjusting by confounder factors, both remained as predictive independent factors for diagnosis of drowning (p = 0.010 and p = 0.022, respectively). We constructed ROC curves for biomarkers' levels attending at the origin of death and established an ApoA1 cut-off point of 100 mg/dL. Correct classification based on the diagnosis criteria was reached for 73.9% of the cases in a discriminant analysis. We propose apolipoprotein A1 (with our cutoff value for correct classification) and α-1 antitrypsin as valuable biomarkers of drowning. Our study, based on forensic cases, reveals our proteomic approach as a new complementary tool in the forensic diagnosis of drowning and, perhaps, in clinical future implications in drowned patients. However, this is a pilot approach, and future studies are necessary to consolidate our promising preliminary data.

Do haematopoietic stem cells age?

Genetic defects that accumulate in haematopoietic stem cells (HSCs) are thought to be responsible for age-related changes in haematopoiesis that include a decline in lymphopoiesis and skewing towards the myeloid lineage. This HSC-centric view is based largely on studies showing that HSCs from aged mice exhibit these lineage biases following transplantation into irradiated young recipient mice. In this Opinion article, we make the case that the reliance on this approach has led to inaccurate conclusions regarding the effects of ageing on blood-forming stem cells; we suggest instead that changes in the environment contribute to haematopoietic system ageing. We propose that a complete understanding of how ageing affects haematopoiesis depends on the analysis of blood cell production in unperturbed mice. We describe how this can be achieved using in situ fate mapping. This approach indicates that changes in downstream progenitors, in addition to any HSC defects, may explain the reduced lymphopoiesis and sustained myelopoiesis that occur during ageing.

Plasma Cells Are Obligate Effectors of Enhanced Myelopoiesis in Aging Bone Marrow.

Aging results in increased myelopoiesis, which is linked to the increased incidence of myeloid leukemias and production of myeloid-derived suppressor cells. Here, we examined the contribution of plasma cells (PCs) to age-related increases in myelopoiesis, as PCs exhibit immune regulatory function and sequester in bone marrow (BM). PC number was increased in old BM, and they exhibited high expression of genes encoding inflammatory cytokines and pathogen sensors. Antibody-mediated depletion of PCs from old mice reduced the number of myeloid-biased hematopoietic stem cells and mature myeloid cells to levels in young animals, but lymphopoiesis was not rejuvenated, indicating that redundant mechanisms inhibit that process. PCs also regulated the production of inflammatory factors from BM stromal cells, and disruption of the PC-stromal cell circuitry with inhibitors of the cytokines IL-1 and TNF-α attenuated myelopoiesis in old mice. Thus, the age-related increase in myelopoiesis is driven by an inflammatory network orchestrated by PCs.

Experiences with Prescribing Large Quantities of Systemic Anticancer Therapy Near Death.

Background: The most important decision after diagnosing terminal cancer is whether to provide active therapy or withhold treatment. Objective: To analyze the aggressiveness of care by evaluating systemic anticancer therapy (SACT) given near to death, describing this care and identifying factors that determine its use. Design: This involves retrospective observational cohorts study. Setting/Subjects: This involves patients with metastatic tumors who died at a University Hospital in Spain between 2015 and 2016. Measurements: Data obtained from prescribing oncologists and patients' clinical records, type of cancer, and information on treatment. The dependent variable used was the interval between the date of the last dose and date of death. Results: Ninety-four (32.60%) of 288 patients received SACT in the last month of life. This cohort had a higher frequency of lung cancer (OR: 1.58; CI 95%: 1.14-2.18), received more care from oncologist 2 (OR: 1.50; CI 95%: 1.08-2.08), had fewer last-line treatment cycles (OR: 1.28; CI 95%: 1.13-1.45), a lower subjective response (OR: 3.13; CI 95%: 1.34-7.29), less clinical benefit (OR: 2.38; CI 95%: 1.04-5.55), more visits to the Emergency Department (OR: 1.59; CI 95%: 1.06-2.38), and less care from the Palliative Care Unit (OR: 4.55; CI 95%: 2.69-7.70). In multivariate analysis, the predictors of having received SACT close to death remained: receiving fewer cycles of treatment (OR: 1.28; CI 95%: 1.12-1.47) and less palliative care (OR: 4.54; CI 95%: 2.56-7.69). Conclusions: A third of cancer patients received SACT in the last month of life with less efficacy and poorer quality of care than patients not receiving it.

Lymphoid-Biased Hematopoietic Stem Cells Are Maintained with Age and Efficiently Generate Lymphoid Progeny.

Current models propose that reductions in the number of lymphoid-biased hematopoietic stem cells (Ly-HSCs) underlie age-related declines in lymphopoiesis. We show that Ly-HSCs do not decline in number with age. Old Ly-HSCs exhibit changes in gene expression and a myeloid-biased genetic profile, but we demonstrate that they retain normal lymphoid potential when removed from the old in vivo environment. Additional studies showing that interleukin-1 inhibits Ly-HSC lymphoid potential provide support for the hypothesis that increased production of inflammatory cytokines during aging underlies declines in lymphocyte production. These results indicate that current models proposing that lymphopoiesis declines with age due to loss of Ly-HSCs require revision and provide an additional perspective on why lymphocyte development in the elderly is attenuated.

Infectious crystalline keratopathy: Management of three cases with different risk factors.

We present the management of three cases of infectious crystalline keratopathy. The first one, in a 46-year-old patient with two previous penetrating keratoplasties; the second one, in a 46-year-old patient with chronic alcoholism and limbal insufficiency; and the third one, in a 70-year-old patient with bullous keratopathy. Other systemic conditions that may mimic infectious crystalline keratopathy, such as multiple myeloma, gout or cystinosis were ruled out on each patient by laboratory testing. The cases were managed with topical or topical and systemic treatment that led to the disappearance of the symptoms. Infectious crystalline keratopathy is a chronic and indolent pathology in which interlamellar bacterial plaques are observed in absence of apparent ocular inflammatory signs. Microorganisms penetrate the cornea through epithelial defects, commonly after a penetrating keratoplasty, although other risk factors may be present.

Cost minimization analysis for basic life support.

BACKGROUND: The use of online teaching methodology for basic life support (BLS) courses is progressively increasing. OBJECTIVE: The objective of this study was to verify whether the blended-learning methodology (virtual course with a short face-to-face complement) was more efficient than a course that followed the classical or face-to-face methodology in our university. MATERIALS AND METHODS: A cost minimization analysis was performed for two BLS and automatic external defibrillation (AED) courses, one of which was conducted face-to-face (Control Group) and the second of which was conducted via blended-learning (Experimental Group). The courses had the same duration and content according to the European Resuscitation Council (ERC) recommendations. In the face-to-face course, direct costs were considered those generated by the faculty and derived from the academic activity. Other costs were those generated by the use of classrooms and the amortization of manikins and AED training. The perspective of the analysis was that of the provider, the academic, and a time horizon of six months. The costs are expressed in € 2017. RESULTS: The savings of a course in BLS-AED based on the blended-learning methodology calculated for a total of 160 university nursing and medical students were € 2328.8 for the first year of its implementation and € 9048.8 for its second edition compared with the same course using a face-to-face methodology. CONCLUSIONS: The blended-learning methodology supposes a cost savings for BLS-AED courses, mainly due to the reduction of expenses of the teaching staff. The blended-learning methodology seems to be more efficient than the face-to-face methodology.