Enhancer-instructed epigenetic landscape and chromatin compartmentalization dictate a primary antibody repertoire protective against specific bacterial pathogens.

Antigen receptor loci are organized into variable (V), diversity (D) and joining (J) gene segments that rearrange to generate antigen receptor repertoires. Here, we identified an enhancer (E34) in the murine immunoglobulin kappa (Igk) locus that instructed rearrangement of Vκ genes located in a sub-topologically associating domain, including a Vκ gene encoding for antibodies targeting bacterial phosphorylcholine. We show that E34 instructs the nuclear repositioning of the E34 sub-topologically associating domain from a recombination-repressive compartment to a recombination-permissive compartment that is marked by equivalent activating histone modifications. Finally, we found that E34-instructed Vκ-Jκ rearrangement was essential to combat Streptococcus pneumoniae but not methicillin-resistant Staphylococcus aureus or influenza infections. We propose that the merging of Vκ genes with Jκ elements is instructed by one-dimensional epigenetic information imposed by enhancers across Vκ and Jκ genomic regions. The data also reveal how enhancers generate distinct antibody repertoires that provide protection against lethal bacterial infection.

A B-Cell-Specific Enhancer Orchestrates Nuclear Architecture to Generate a Diverse Antigen Receptor Repertoire.

The genome is organized into topologically associated domains (TADs) that enclose smaller subTADs. Here, we identify and characterize an enhancer that is located in the middle of the V gene region of the immunoglobulin kappa light chain (Igκ) locus that becomes active preceding the stage at which this locus undergoes V(D)J recombination. This enhancer is a hub of long-range chromatin interactions connecting subTADs in the V gene region with the recombination center at the J genes. Deletion of this element results in a highly altered long-range chromatin interaction pattern across the locus and, importantly, affects individual V gene utilization locus-wide. These results indicate the existence of an enhancer-dependent framework in the Igκ locus and further suggest that the composition of the diverse antibody repertoire is regulated in a subTAD-specific manner. This enhancer thus plays a structural role in orchestrating the proper folding of the Igκ locus in preparation for V(D)J recombination.

Enhancers within the Ig V Gene Region Orchestrate Chromatin Topology and Regulate V Gene Rearrangement Frequency to Shape the B Cell Receptor Repertoire Specificities.

Effective Ab-mediated responses depend on a highly diverse Ab repertoire with the ability to bind a wide range of epitopes in disease-causing agents. The generation of this repertoire depends on the somatic recombination of the variable (V), diversity (D), and joining (J) genes in the Ig loci of developing B cells. It has been known for some time that individual V, D, and J gene segments rearrange at different frequencies, but the mechanisms behind this unequal V gene usage have not been well understood. However, recent work has revealed that newly described enhancers scattered throughout the V gene-containing portion of the Ig loci regulate the V gene recombination frequency in a regional manner. Deletion of three of these enhancers revealed that these elements exert many layers of control during V(D)J recombination, including long-range chromatin interactions, epigenetic milieu, chromatin accessibility, and compartmentalization.

The branched mitochondrial respiratory chain from the jellyfish Stomolophus sp2 as a probable adaptive response to environmental changes.

During their long evolutionary history, jellyfish have faced changes in multiple environmental factors, to which they may selectively fix adaptations, allowing some species to survive and inhabit diverse environments. Previous findings have confirmed the jellyfish's ability to synthesize large ATP amounts, mainly produced by mitochondria, in response to environmental challenges. This study characterized the respiratory chain from the mitochondria of the jellyfish Stomolophus sp2 (previously misidentified as Stomolophus meleagris). The in-gel activity from isolated jellyfish mitochondria confirmed that the mitochondrial respiratory chain contains the four canonical complexes I to IV and F0F1-ATP synthase. Specific additional activity bands, immunodetection, and mass spectrometry identification confirmed the occurrence of four alternative enzymes integrated into a branched mitochondrial respiratory chain of Stomolophus sp2: an alternative oxidase and three dehydrogenases (two NADH type II enzymes and a mitochondrial glycerol-3-phosphate dehydrogenase). The analysis of each transcript sequence, their phylogenetic relationships, and each protein's predicted models confirmed the mitochondrial alternative enzymes' identity and specific characteristics. Although no statistical differences were found among the mean values of transcript abundance of each enzyme in the transcriptomes of jellyfish exposed to three different temperatures, it was confirmed that each gene was expressed at all tested conditions. These first-time reported enzymes in cnidarians suggest the adaptative ability of jellyfish's mitochondria to display rapid metabolic responses, as previously described, to maintain energetic homeostasis and face temperature variations due to climate change.

Real-life analysis on safety and efficacy of asciminib for ponatinib pretreated patients with chronic myeloid leukemia.

Failure of second-generation tyrosine kinase inhibitors (2GTKI) is a challenging situation in patients with chronic myeloid leukemia (CML). Asciminib, recently approved by the US Federal Drug Administration, has demonstrated in clinical trials a good efficacy and safety profile after failure of 2GTKI. However, no study has specifically addressed response rates to asciminib in ponatinib pretreated patients (PPT). Here, we present data on responses to asciminib from 52 patients in clinical practice, 20 of them (38%) with prior ponatinib exposure. We analyzed retrospectively responses and toxicities under asciminib and compared results between PPT and non-PPT patients.After a median follow-up of 30 months, 34 patients (65%) switched to asciminib due to intolerance and 18 (35%) due to resistance to prior TKIs. Forty-six patients (88%) had received at least 3 prior TKIs. Regarding responses, complete cytogenetic response was achieved or maintained in 74% and 53% for non-PPT and PPT patients, respectively. Deeper responses such as major molecular response and molecular response 4.5 were achieved in 65% and 19% in non-PPT versus 32% and 11% in PPT, respectively. Two patients (4%) harbored the T315I mutation, both PPT.In terms of toxicities, non-PPT displayed 22% grade 3-4 TEAE versus 20% in PPT. Four patients (20% of PPT) suffered from cross-intolerance with asciminib as they did under ponatinib.Our data supports asciminib as a promising alternative in resistant and intolerant non-PPT patients, as well as in intolerant PPT patients; the resistant PPT subset remains as a challenging group in need of further therapeutic options.

Variable endothelial cell function restoration after initiation of two antiretroviral regimens in HIV-infected individuals.

Background: The effect of ART on endothelial cell function is incompletely characterized. Methods: We performed a 24 week prospective, case-control and comparative pilot study of ART-naive HIV-infected patients who started a darunavir- or rilpivirine-based regimen, matched with non-HIV-infected volunteers, to compare changes at week 24 from baseline in levels of circulating endothelial cells (CECs), endothelial progenitor cells (EPCs) and circulating angiogenic cells, as well as changes in immune-activation markers. Results: The study population comprised 24 HIV-infected patients and 24 non-infected volunteers. Both HIV groups completely suppressed viraemia. HIV-infected patients had higher levels of activation markers than the control group in CD8 T cells at baseline; these decreased after 24 weeks of treatment, but without reaching the levels of the control group. No statistical differences in immune activation were seen between the darunavir and rilpivirine groups. Levels of CECs were higher and levels of EPCs and circulating angiogenic cells were lower in HIV-infected patients than in the control group, although these parameters were similar between the darunavir group and the control group, but not the rilpivirine group, at week 24. An unfavourable association was observed between rilpivirine, age and increased number of CECs. Conclusions: Restoration of circulating levels of EPCs and CECs in darunavir-treated patients was greater than in those treated with rilpivirine, suggesting ongoing endothelial repair mechanisms.

Because difficulty is not the same for everyone: the impact of complexity in working memory is associated with cannabinoid 1 receptor genetic variation in young adults.

Individual differences in working memory ability are mainly revealed when a demanding challenge is imposed. Here, we have associated cannabinoid 1 (CB1) receptor genetic variation rs2180619 (AA, AG, GG), which is located in a potential CNR1 regulatory sequence, with performance in working memory. Two-hundred and nine Mexican-mestizo healthy young participants (89 women, 120 men, mean age: 23.26 years, SD = 2.85) were challenged to solve a medium (2-back) vs. a high (3-back) difficulty N-back tasks. All subjects responded as expected, performance was better with the medium than the high demand task version, but no differences were found among genotypes while performing each working memory (WM) task. However, the cost of the level of complexity in N-back paradigm was double for GG subjects than for AA subjects. It is noteworthy that an additive-dosage allele relation was found for G allele in terms of cost of level of complexity. These genetic variation results support that the endocannabinoid system, evaluated by rs2180619 polymorphism, is involved in WM ability in humans.

A preliminary longitudinal study on the cognitive and functional outcome of bipolar excellent lithium responders.

BACKGROUND: Neurocognitive dysfunction in bipolar disorder represents a possible marker of underlying pathophysiology, but to date, most studies are cross-sectional and heterogeneous with regard to pharmacological treatments. In the present study we investigated the 6-year cognitive and functional outcome of a sample of euthymic excellent lithium responders (ELR). METHOD: A total sample of twenty subjects was assessed at baseline and 6years later: ten diagnosed of bipolar disorder according to DSM-IV criteria and ten healthy matched controls. The sample size was enough to find statistical differences between groups, with a statistical power of 0.8. Bipolar patients were on lithium treatment during all this follow-up period and fulfilled ELR criteria as measured by the Alda scale. A neuropsychological test battery tapping into the main cognitive domains was used at baseline and at after 6-year of follow-up. Functional outcome was evaluated by means of the Functioning Assessment Short Test at study endpoint. RESULTS: Repeated measures multivariate analyses of variance showed that bipolar patients were cognitively impaired in the executive functioning, inhibition, processing speed and verbal memory domains (p<0.03) compared to controls and such deficits were stable over time. Longer duration of illness and lower psychosocial outcome were significantly related to cognitive impairment (p<0.05). CONCLUSIONS: Cognitive dysfunction was present even in euthymic ELR. These deficits remain stable over the long term, and are basically associated with greater symptoms and poorer psychosocial adjustment.

Role of the clinical radiopharmacist in patient safety during myocardial perfusion imaging with vasodilator stress agents.

AIM: To assess the radiopharmacist's role in a multidisciplinary team focused on the contraindications of regadenoson in order to ensure the safe use of pharmacologic vasodilator stress agents in patients undergoing SPECT-MPI. METHODS: We ambispectively studied its safe use in 1905 patients (54.1% female, mean age: 66.6±11.7 years, range: 20-95 years). Sex, age, medical history, medications, drug allergies, and contraindications for stress testing were registered together with recommendations for the nuclear physician in charge. RESULTS: Detected contraindications and corresponding recommendations were as follows: risk factors for QTc interval prolongation 7.5% - measurement of QTc interval previously to test and monitor ECG; prior stroke or TIA 4.2% - consider carotid stenosis assessment; salicylates/sulfonamides allergy 3.1% - use 99mTc-sestamibi; epilepsy or risk factors for seizures 2.4% - use of adenosine or reconsider test indication; systemic corticosteroid therapy for severe COPD 1.3% - reassessment of patient's condition; acute exacerbation of COPD 0.8% - defer test until acute episode is over; severe asthma 0.4% - do not perform test; methylxanthine ingestion 0.3% - avoid consumption previously; other 6.1% - evaluation of other contraindications. No contraindications were detected in 73.6% of patients. The test was canceled due to absolute contraindications in 2.9% of the requests. CONCLUSIONS: Working in a systematic way, the radiopharmacist was able to detect a high number of issues related to regadenoson, with one out of four patients presenting some clinical contraindication. The recommendations given by the radiopharmacist were well accepted by the nuclear physicians who changed their approach contributing to increase the safety of patients referred for MPI.

Persistence of cognitive impairment and its negative impact on psychosocial functioning in lithium-treated, euthymic bipolar patients: a 6-year follow-up study.

BACKGROUND: Previous cross-sectional studies report that cognitive impairment is associated with poor psychosocial functioning in euthymic bipolar patients. There is a lack of long-term studies to determine the course of cognitive impairment and its impact on functional outcome. Method A total of 54 subjects were assessed at baseline and 6 years later; 28 had DSM-IV TR bipolar I or II disorder (recruited, at baseline, from a Lithium Clinic Program) and 26 were healthy matched controls. They were all assessed with a cognitive battery tapping into the main cognitive domains (executive function, attention, processing speed, verbal memory and visual memory) twice over a 6-year follow-up period. All patients were euthymic (Hamilton Rating Scale for Depression score lower than 8 and Young mania rating scale score lower than 6) for at least 3 months before both evaluations. At the end of follow-up, psychosocial functioning was also evaluated by means of the Functioning Assessment Short Test. RESULTS: Repeated-measures multivariate analysis of covariance showed that there were main effects of group in the executive domain, in the inhibition domain, in the processing speed domain, and in the verbal memory domain (p<0.04). Among the clinical factors, only longer illness duration was significantly related to slow processing (p=0.01), whereas strong relationships were observed between impoverished cognition along time and poorer psychosocial functioning (p<0.05). CONCLUSIONS: Executive functioning, inhibition, processing speed and verbal memory were impaired in euthymic bipolar out-patients. Although cognitive deficits remained stable on average throughout the follow-up, they had enduring negative effects on psychosocial adaptation of patients.

[Nuclear Cardiology in the COVID-19 pandemic]. / La cardiología nuclear en la pandemia COVID-19.

SARS-CoV-2 infection has a very important relationship with cardiovascular disease. Since the beginning of the pandemic, a close relationship has been observed between cardiovascular comorbidity and a worse prognosis in COVID-19 patients. The study of the pathophysiology of SARS-CoV-2 infection and cardiovascular disease suggests several concomitant hypotheses: direct myocardial damage by the virus, hypoxemia secondary to respiratory failure, inflammatory response to infection and/or thromboembolic phenomena. Cardiovascular damage can manifest in the acute phase of infection with acute myocardial infarction, myocarditis, arrhythmias..., during this phase Nuclear Cardiology procedures have not played a determining role in the diagnosis and management of these patients. On the other hand, in the subacute phase of the infection and in the post-acute COVID syndrome, Nuclear Cardiology seems to shed light on what happens in the cardiovascular system in this phase of the disease.The COVID-19 pandemic has represented a great challenge for health systems, with a significant reduction in non-urgent diagnostic procedures with the aim of reducing the risk of transmission to patients and health personnel. Nuclear Cardiology has not been an exception. In addition to the prioritization of urgent/non-deferrable procedures and general screening, hygiene and distance measures, the main organizations and scientific societies of Nuclear Medicine and Nuclear Cardiology released recommendations and guidelines for safe practice, introducing significant changes in myocardial perfusion SPECT protocols.

Assessment of pulmonary embolism related to active SARS-CoV-2 infection in pregnant women.

AIM: To analyze a sample of pregnant patients who underwent pulmonary perfusion scintigraphy to rule out suspicion of pulmonary embolism (PE) during the acute COVID-19 infection hospitalization period in our hospital. MATERIAL AND METHODS: SPECT scintigraphy with a reduced dose (111MBq) of 99mTc-macroaggregated albumin was performed in all the patients (n=5). The images obtained were interpreted by comparing the findings with the radiological images according to the PISAPED criteria. RESULTS: Only one of the 5 patients was diagnosed with PE. Two patients showed pathological scintigraphy findings attributable to radiological alterations due to COVID-19 pneumonia, and the other two had normal pulmonary perfusion. CONCLUSION: Given the non-specific features of the clinical manifestations and d-dimer values ​​in COVID-19, as well as their similarity to those of PE, pulmonary perfusion scintigraphy plays a crucial role in the screening of PE in these patients due to its high sensitivity and lower irradiation compared to CT. Despite the limited number of patients, the results obtained have special relevance due to the absence of scientific publications in this group of patients within the exceptional context of the COVID-19 pandemic.

Incidence and prognostic impact of U2AF1 mutations and other gene alterations in myelodysplastic neoplasms with isolated 20q deletion.

BACKGROUND: In myelodysplastic neoplasms (MDS), the 20q deletion [del(20q)] is a recurrent chromosomal abnormality that it has a high co-occurrence with U2AF1 mutations. Nevertheless, the prognostic impact of U2AF1 in these MDS patients is uncertain and the possible clinical and/or prognostic differences between the mutation type and the mutational burden are also unknown. METHODS: Our study analyzes different molecular variables in 100 MDS patients with isolated del(20q). RESULTS & CONCLUSIONS: We describe the high incidence and negative prognostic impact of U2AF1 mutations and other alterations such as in ASXL1 gene to identify prognostic markers that would benefit patients to receive earlier treatment.

Preload dependence of gated cardiac SPECT-derived ventricular volumes in hemodialysis patients.

BACKGROUND: In hemodialysis (HD) patients, the intravascular volume expansion of the pre-HD state leads to a high preload. We aim to examine its effect on myocardial perfusion gated SPECT (MPGS)-derived left ventricular (LV) volumes. METHODS: The study comprised 50 end-stage renal disease (ESRD) patients on HD with normal 2-day stress/rest MPGS performed for kidney transplantation risk assessment. Patients (pts) comprised 23 men/27 women, with mean age of 59.4 ± 7.1 years. The time elapsed from the last HD session in hours was calculated on both days, and patients were classified according to whether it was higher (group A: 19 pts), lower (group B: 27 pts), or equal (group C: 4 pts) on the stress vs the rest day. End-diastolic, end-systolic volumes (EDV, ESV), and left ventricular ejection fraction (LVEF) were determined using QGS™ software. Transient ischemic dilation (TID) ratios were derived from the nongated images using QPS™ software. RESULTS: Volumes were significantly higher at stress in group A, at rest in group B, and similar in group C. TID ratios were significantly higher in group A vs groups B and C. CONCLUSIONS: MPGS-derived ventricular volumes are preload dependent. The high preload of the pre-HD state may mimic ischemic TID if occurring on the stress day and create confusion if coinciding with the rest day.

Postnatal maturation of primary auditory cortex in the mustached bat, Pteronotus parnellii.

The primary auditory cortex (AI) of adult Pteronotus parnellii features a foveal representation of the second harmonic constant frequency (CF2) echolocation call component. In the corresponding Doppler-shifted constant frequency (DSCF) area, the 61 kHz range is over-represented for extraction of frequency-shift information in CF2 echoes. To assess to which degree AI postnatal maturation depends on active echolocation or/and reflects ongoing cochlear maturation, cortical neurons were recorded in juveniles up to postnatal day P29, before the bats are capable of active foraging. At P1-2, neurons in posterior AI are tuned sensitively to low frequencies (22-45 dB SPL, 28-35 kHz). Within the prospective DSCF area, neurons had insensitive responses (>60 dB SPL) to frequencies <40 kHz and lacked sensitive tuning curve tips. Up to P10, when bats do not yet actively echolocate, tonotopy is further developed and DSCF neurons respond to frequencies of 51-57 kHz with maximum tuning sharpness (Q(10dB)) of 57. Between P11 and 20, the frequency representation in AI includes higher frequencies anterior and dorsal to the DSCF area. More multipeaked neurons (33%) are found than at older age. In the oldest group, DSCF neurons are tuned to frequencies close to 61 kHz with Q(10dB) values < or =212, and threshold sensitivity, tuning sharpness and cortical latencies are adult-like. The data show that basic aspects of cortical tonotopy are established before the bats actively echolocate. Maturation of tonotopy, increase of tuning sharpness, and upward shift in the characteristic frequency of DSCF neurons appear to strongly reflect cochlear maturation.

Dynamics of dermatophytosis frequency in Mexico: an analysis of 2084 cases.

We analysed 15,101 biological samples from patients presenting with superficial mycoses who attended outpatient services over a 10-year period. Scale samples were processed for direct microscopic examination with 15% KOH and cultured on Sabouraud glucose agar plus chloramphenicol and cycloheximide. Laboratory examination confirmed 4,709 cases of superficial mycosis (31.18%), of which 2,084 (44.26%) were dermatophytoses. The species most frequently encountered was Trichophyton rubrum (71.2%), followed by T. tonsurans (6.9%), T. mentagrophytes (5.5%), Microsporum canis (4.5%) and Epidermophyton floccosum (1.3%). The most frequent clinical form of dermatophytosis was tinea unguium (59.9%), followed by tinea pedis (24.5%). We demonstrate that the number of cases of T. rubrum is increasing in Mexico.

[Occupational health practice among law, evidence and ethics: a field study]. / La pratica professionale del medico del lavoro tra norme, prove scientifiche ed etica professionale: un'esperienza sul campo.

BACKGROUND: Quality in health care is defined as the degree to which health services increase the likelihood of desired health outcomes and are consistent with the law, the evidence and the ethical values. MATERIALS AND METHODS: The paper aims at providing a framework in which the concept of quality is applied in the field of occupational health by analysing real-life decisions. RESULTS: The Italian law states that the practice must be evidence-based and respectful of the code of ethics, emphasizeing that the practice should be relevant, knowledge-based, sound and appropriate. Any dilemma should be dealt according to the ethical principles of beneficience, autonomy and justice. Both the law and the code require that the practice is based on the available evidence. Results show that decisions in selected cases can be made according the above mentioned criteria. CONCLUSION; Decisions made on the basis of a process founded on scientific evidence result in effective and high-quality outcomes respectful of both law and ethics.

[Quantitative salivary gland scintigraphy in head and neck cancer patients following radiotherapy]. / Gammagrafía salival cuantitativa en pacientes con cáncer de cabeza y cuello tratados con radioterapia.

OBJECTIVE: To assess the role of quantitative salivary gland scintigraphy (SGS) in the detection of functional impairment of salivary glands in patients with head and neck cancer treated with radiotherapy (RT). MATERIAL AND METHODS: We studied 19 patients (17 men), mean age 62.4 years (44-75). Three studies were performed to each patient: baseline, 3 and 18 months after RT. SGS was acquired for 25 minutes following injection of 3.7 MBq/kg of (99m)Tc-pertechnetate with lemon juice at the end of minute 15. Excretion fraction (EF), counts per minute/pixel/MBq (CMPM) and uptake percentage were obtained from time-activity curves from ROIs placed over parotid (PG) and submandibular glands (SMG) and related to the doses received by the PG. RESULTS: EF showed a significant reduction from the baseline to the 3 months study (p<0.001) for the PG and SMG and from the baseline to the 18 months study for the SMG (p<0.001). A significant improvement of EF was seen from the 3 months to the 18 months study for the PG (p<0.05). CMPM did not change significantly from the baseline to the 3 months and 18 months studies for the PG and showed a significant reduction (p<0.01) for the SMG from the baseline to 18 months study. The uptake percentage did not change significantly between studies. A moderate association was observed between the doses to PG and the 3m study parameters. CONCLUSIONS: EF was more sensitive than uptake in assessing post-RT impairment of salivary function. In addition, it reflected functional recovery of parotid glands over time.