Cloning and functional characterization of novel human neutralizing anti-interferon-alpha and anti-interferon-beta antibodies.

Type I interferons (IFNs) play a pivotal role in immune response modulation, yet dysregulation is implicated in various disorders. Therefore, it is crucial to develop tools that facilitate the understanding of their mechanism of action and enable the development of more effective anti-IFN therapeutic strategies. In this study, we isolated, cloned, and characterized anti-IFN-α and anti-IFN-ß antibodies (Abs) from peripheral blood mononuclear cells of individuals treated with IFN-α or IFN-ß, harboring confirmed neutralizing Abs. Clones AH07856 and AH07857 were identified as neutralizing anti-IFN-α-specific with inhibition against IFN-α2a, -α2b, and -αK subtypes. Clones AH07859 and AH07866 were identified as neutralizing anti-IFN-ß1a-specific signaling, and able to block Lipopolysaccharide or S100 calcium binding protein A14-induced IFN-ß signaling effects. Cloned Abs bind rhesus but not murine IFNs. The specificity of inhibition between IFN-α and IFN-ß suggests potential for diverse research and clinical applications.

Epidemiological and Financial Aspects of Hospitalizations for Bacterial Meningitis in Brazil.

Introduction: Understanding the epidemiology and cost implications of acute bacterial meningitis is crucial for effective health planning, timely treatment implementation, and comprehensive patient support measures, as well as for determining appropriate hospital expenses. Therefore, we conducted an analysis of hospitalization cases for bacterial meningitis in Brazil from January 2008 to December 2019. Methods: This is a descriptive ecological study that utilized the Hospital Information System of Brazil's National Unified Health System (SIH/SUS) database. The variables included sex, region, age group, hospitalizations, deaths, lethality rate, and hospital service expenses. The data were tabulated to focus specifically on the epidemiological aspect of bacterial meningitis. Results: During the study period, there were 20,207 hospitalizations for bacterial meningitis in Brazil. Men accounted for a higher number of cases, with 11,690 (57.67%), while women had a higher lethality rate of 10.64%. The Southeast region had the highest percentage of both hospitalizations (45.78%) and deaths (46.42%). Bacterial meningitis remains an important cause of morbidity and mortality, particularly in children under 5 years of age. Notably, the elderly and the Northeast region showed higher rates of lethality. The total expenditure on hospital services exceeded 43 million in Brazilian real, with the highest expenditure observed in 2019 and the lowest in 2011. Conclusion: A higher prevalence of the disease was observed in males, in children under 1-year-old and in the southeast region. Hospital expenditures were found to be substantial and increasing over time, underscoring the significance of early diagnosis and the promotion of vaccination campaigns.

Effects of Physical Exercise on Cerebral Blood Velocity in Older Adults: A Systematic Review and Meta-Analysis.

As the older population grows, there is an increasing interest in understanding how physical exercise can counteract the changes seen with aging. The benefits of exercise to general health, and especially to the cardiovascular system, have been a topic of discussion for decades. However, there is still a need to elucidate the effects of training programs on the cerebrovascular blood velocity in older people. This systematic review and meta-analysis aimed to investigate the effect of physical exercise on the cerebral blood velocity in older people (PROSPERO CRD42019136305). A search was performed on PubMed, Web of Science, EBSCO, ScienceDirect, and Scopus from the inception of this study to October 2023, retrieving 493 results, of which 26 were included, analyzing more than 1000 participants. An overall moderate risk of bias was found for the studies using the Cochrane risk-of-bias tools for randomized and non-randomized clinical trials. The pooled results of randomized trials showed that older people who underwent physical exercise presented a statistically significant increase in cerebral blood velocity (3.58; 95%CI = 0.51, 6.65; p = 0.02). This result indicates that physical exercise is important to help maintain cerebral health in older adults.

Middle cerebral artery blood velocity and cognitive function after high- and moderate-intensity aerobic exercise sessions.

This crossover study explored the acute effect of a session of high-intensity interval training (HIIT) or moderate-intensity continuous training (MICT) on middle cerebral artery (MCA) variables such as cerebral blood velocity, pulsatility index (PI) and resistivity index (RI) through transcranial Doppler (TCD), and cognitive function (CF - verbal fluency and Digit Span) in healthy young adults. Participants (26 healthy young adults, 13 women, 24 ± 3 years) underwent two different randomized exercise sessions: (1) MICT (60 % heart rate reserve, HRR) and (2) HIIT (80 % HRR). MCA velocity, PI, RI, CF, and serum lactate were measured immediately before and after the sessions. HIIT demonstrated improved executive function/semantic fluency (20 %, p = 0.019), while both MICT and HIIT increased lactate (625 %, HIIT, p < 0.001, and 238 %, MICT, p < 0.001). Other assessments remained stable, except for reduced PI (p = 0.029) and RI (p = 0.023) after MICT, with no significant difference (pre-post for HIIT-MICT). Notably, cognition improvement correlated with lactate increase in HIIT (ρ = 0.436; p < 0.001). Executive function/semantic fluency increased after HIIT relative to MICT. The findings show that there are no systematic out-of-normal changes in the cerebrovascular circulation of clinically healthy adults undergoing HIIT and MICT.

An African-Specific Variant of TP53 Reveals PADI4 as a Regulator of p53-Mediated Tumor Suppression.

TP53 is the most frequently mutated gene in cancer, yet key target genes for p53-mediated tumor suppression remain unidentified. Here, we characterize a rare, African-specific germline variant of TP53 in the DNA-binding domain Tyr107His (Y107H). Nuclear magnetic resonance and crystal structures reveal that Y107H is structurally similar to wild-type p53. Consistent with this, we find that Y107H can suppress tumor colony formation and is impaired for the transactivation of only a small subset of p53 target genes; this includes the epigenetic modifier PADI4, which deiminates arginine to the nonnatural amino acid citrulline. Surprisingly, we show that Y107H mice develop spontaneous cancers and metastases and that Y107H shows impaired tumor suppression in two other models. We show that PADI4 is itself tumor suppressive and that it requires an intact immune system for tumor suppression. We identify a p53-PADI4 gene signature that is predictive of survival and the efficacy of immune-checkpoint inhibitors. SIGNIFICANCE: We analyze the African-centric Y107H hypomorphic variant and show that it confers increased cancer risk; we use Y107H in order to identify PADI4 as a key tumor-suppressive p53 target gene that contributes to an immune modulation signature and that is predictive of cancer survival and the success of immunotherapy. See related commentary by Bhatta and Cooks, p. 1518. This article is highlighted in the In This Issue feature, p. 1501.

Effects of exercise intervention on psychotic symptoms: A meta-analysis and hypothetical model of neurobiological mechanisms.

There is conflicting evidence on the efficacy of exercise as intervention for psychosis. This article aims to analyze the effect of exercise on psychotic symptoms. A database search was conducted in PubMed, Web of Science, Scopus, ScienceDirect, EBSCO and Cochrane CENTRAL, based on a protocol (PROSPERO: CRD42022326944). Papers available by March 2023 assessing exercise interventions in psychotic patients were included. A significant improvement was found in Positive and Negative Syndrome Scale (PANSS) positive symptoms (MD = -0.75 [-1.35, -0.15], p = 0.01), with large effect sizes for PANSS-negative and general symptoms (-2.14 [-3.36, -0.92]) and (-2.53 [-3.15, -1.91]), respectively. Heterogeneity was high among studies, 49 and 73% for PANSS-positive and negative symptoms, and low, 0%, for general symptoms. It was hypothesized that functioning of specific brain areas, such as the temporal lobe and hippocampus, may underlie the improvement seen with exercise. Based on neuroimaging/neurophysiology studies, we propose a neurobiological model accounting for the association between exercise and psychotic symptom improvement.

Does acceptance-based treatment moderate the effect of stress on dietary lapses?

Weight loss interventions are successful to the extent that participants adhere to calorie-reduced dietary prescriptions and may be undermined by dietary lapses triggered by external or internal factors, such as stress. The purpose of this study was to examine whether an acceptance-based treatment (ABT) versus a standard behavioral treatment (SBT) moderated the effect of stress on dietary lapses. Ecological momentary assessment data were collected from 189 participants who were randomly assigned to either ABT or SBT. Between-subject and within-subject stress were used to predict lapse occurrence along with intervention condition, and the interaction between intervention condition and stress using a generalized linear mixed-effects model. Gender, time of day, and baseline mean lapses/day were included as control variables. Higher mean baseline lapses/day, female gender, and later time of day were significantly associated with increased odds of lapse. Between-subject stress was positively and significantly related to the odds of lapse. Compared to SBT, ABT showed a significant decrease in the odds of lapse at an individual's average level of within-subject stress, but the interaction of intervention condition with within-subject stress was not statistically significant. Participants with high overall stress levels lapsed more frequently than those with low overall stress levels. There was a significant decrease in the odds of lapse for the ABT group at a participant's own average stress level, suggesting that ABT may be beneficial at participants' usual stress levels but when they deviate from their usual stress level, there are no treatment differences.

SARS-CoV-2 Envelope (E) protein interacts with PDZ-domain-2 of host tight junction protein ZO1.

Newly emerged SARS-CoV-2 is the cause of an ongoing global pandemic leading to severe respiratory disease in humans. SARS-CoV-2 targets epithelial cells in the respiratory tract and lungs, which can lead to amplified chloride secretion and increased leak across epithelial barriers, contributing to severe pneumonia and consolidation of the lungs as seen in many COVID-19 patients. There is an urgent need for a better understanding of the molecular aspects that contribute to SARS-CoV-2-induced pathogenesis and for the development of approaches to mitigate these damaging pathologies. The multifunctional SARS-CoV-2 Envelope (E) protein contributes to virus assembly/egress, and as a membrane protein, also possesses viroporin channel properties that may contribute to epithelial barrier damage, pathogenesis, and disease severity. The extreme C-terminal (ECT) sequence of E also contains a putative PDZ-domain binding motif (PBM), similar to that identified in the E protein of SARS-CoV-1. Here, we screened an array of GST-PDZ domain fusion proteins using either a biotin-labeled WT or mutant ECT peptide from the SARS-CoV-2 E protein. Notably, we identified a singular specific interaction between the WT E peptide and the second PDZ domain of human Zona Occludens-1 (ZO1), one of the key regulators of TJ formation/integrity in all epithelial tissues. We used homogenous time resolve fluorescence (HTRF) as a second complementary approach to further validate this novel modular E-ZO1 interaction. We postulate that SARS-CoV-2 E interacts with ZO1 in infected epithelial cells, and this interaction may contribute, in part, to tight junction damage and epithelial barrier compromise in these cell layers leading to enhanced virus spread and severe dysfunction that leads to morbidity. Prophylactic/therapeutic intervention targeting this virus-host interaction may effectively reduce airway and/or gastrointestinal barrier damage and mitigate virus spread.

SARS-CoV-2 Envelope (E) Protein Interacts with PDZ-Domain-2 of Host Tight Junction Protein ZO1.

Newly emerged SARS-CoV-2 is the cause of an ongoing global pandemic leading to severe respiratory disease in humans. SARS-CoV-2 targets epithelial cells in the respiratory tract and lungs, which can lead to amplified chloride secretion and increased leak across epithelial barriers, contributing to severe pneumonia and consolidation of the lungs as seen in many COVID-19 patients. There is an urgent need for a better understanding of the molecular aspects that contribute to SARS-CoV-2 induced pathogenesis and for the development of approaches to mitigate these damaging pathologies. The multifunctional SARS-CoV-2 Envelope (E) protein contributes to virus assembly/egress, and as a membrane protein, also possesses viroporin channel properties that may contribute to epithelial barrier damage, pathogenesis, and disease severity. The extreme C-terminal (ECT) sequence of E also contains a putative PDZ-domain binding motif (PBM), similar to that identified in the E protein of SARS-CoV-1. Here, we screened an array of GST-PDZ domain fusion proteins using either a biotin-labeled WT or mutant ECT peptide from the SARS-CoV-2 E protein. Notably, we identified a singular specific interaction between the WT E peptide and the second PDZ domain of human Zona Occludens-1 (ZO1), one of the key regulators of TJ formation/integrity in all epithelial tissues. We used homogenous time resolve fluorescence (HTRF) as a second complementary approach to further validate this novel modular E-ZO1 interaction. We postulate that SARS-CoV-2 E interacts with ZO1 in infected epithelial cells, and this interaction may contribute, in part, to tight junction damage and epithelial barrier compromise in these cell layers leading to enhanced virus spread and severe respiratory dysfunction that leads to morbidity. Prophylactic/therapeutic intervention targeting this virus-host interaction may effectively reduce airway barrier damage and mitigate virus spread.

Assessment of the non-surgical treatment of patients with rectal cancer who underwent neoadjuvant treatment with chemotherapy and radiotherapy at the oncology department / Avaliação do tratamento não cirúrgico de pacientes com câncer de reto submetido ao tratamento neoadjuvante com quimioterapia e radioterapia no serviço de oncologia

ABSTRACT Objective: To describe the partial results of a study in patients with rectal cancer who underwent neoadjuvant treatment with chemotherapy and radiotherapy regarding the rate of complete clinical response, disease-free survival, anorectal function, and quality of life. Material and methods: This was a prospective study from June 2015 to June 2018, in patients with low- or mid-rectum adenocarcinoma and clinical stage II or III, treated with radiotherapy and chemotherapy (IMRT 54 Gy for six weeks) concomitant with 5-fluorouracil (5-FU) 380 mg/m2 and folinic acid (LV) 20 mg/m2 for five days in the first and fifth weeks and two cycles after radiotherapy (5-FU 400 mg/m2 and LV 20 mg/m2) every 28 days. After the treatment, clinical examination, rectosigmoidoscopy, pelvic magnetic resonance imaging, chest and upper abdomen computed tomography, and CEA testing were performed. Resection surgery was performed in those with incomplete clinical response (iCR). Those with complete clinical response (cCR) are under observation (wait-and-see policy). Manometry and scintigraphic function and quality of life scales were collected before treatment and at 30 and 90 days after the end of treatment. Results: As of June 2018, 11 patients were recruited. One was excluded from the analysis for presenting severe toxicity, suggestive of dihydropyrimidine dehydrogenase (DPD) deficiency, after the first chemotherapy cycle. All others completed the treatment. Two patients presented toxicity grade 3/4 related to chemotherapy and had their doses reduced. Seven patients (70%) presented iRC; three underwent rectosigmoidectomy, and the anatomopathological evaluation indicated complete pathological response in two cases (28.5%). Three (30%) presented cCR and did not present evidence of disease after a mean follow-up of 19 months. Patients presented improvement of anorectal function and quality of life. Conclusions: Advances in the neoadjuvant treatment of rectal tumors contributed to better rates of complete pathological responses. New paradigms promote an increase in the complete clinical response rates, which would allow organ preservation and consequent reduction of surgical morbidity.

RESUMO Objetivo: Descrever os resultados parciais de estudo em pacientes com câncer de reto submetidos a tratamento neoadjuvante com quimioterapia e radioterapia quanto à taxa resposta clínica completa, sobrevida livre de doença, função anorretal e qualidade de vida. Material e métodos: Estudo prospectivo desde junho 2015 até junho de 2018, em paciente com adenocarcinoma de reto baixo ou médio e estadio clínico II ou III tratados com RT/QT (IMRT 54 Gy em 6 semanas) concomitante a 5-Fuorouracil (5-FU) 380 mg/m2 e ácido folínico (LV) 20 mg/m2 por 5 dias nas primeira e quinta semanas e dois ciclos após RT (5-FU 400 mg/m2 e LV 20 mg/m2) a cada 28 dias. Após o tratamento, realizou-se exame clínico, retossigmoidoscopia, RNM de pelve, TC de tórax e abdômen superior e dosagem de CEA. Naqueles com Resposta Clínica Incompleta (iRC) procedeu-se à cirurgia de ressecção. Aqueles com Resposta Completa (cRC) estão em observação (wait and see policy). Manometria e escalas de função esfincteriana e qualidade de vida foram obtidas antes e após 30 e 90 dias do término do tratamento. Resultados: Até junho de 2018, recrutaram-se 11 pacientes. Um foi excluído da análise, pois apresentou toxicidade severa sugestiva de deficiência de DPD após o primeiro ciclo de QT. Todos os demais concluíram o tratamento. Toxicidades graus 3/4 relacionadas à QT ocorreram dois pacientes, reduzindo-se sua dose. Sete (70%) apresentaram iRC, submetendo três à retossigmoidectomia cuja avaliação anatomopatológica evidenciou Resposta Completa (pRC) em dois casos (28,5%). Três (30%) apresentaram cRC e estão sem evidência de doença com seguimento médio de 19 meses. Houve melhora da função anorretal e da qualidade de vida. Conclusões: Avanços no tratamento neoadjuvante dos tumores de reto contribuíram para melhores taxas de pRC. Novos paradigmas promovem crescentes índices de cRC, o que possibilitaria a preservação do órgão e consequente redução da morbidade cirúrgica.

Dysregulation of synaptogenesis genes antecedes motor neuron pathology in spinal muscular atrophy.

The motor neuron (MN) degenerative disease, spinal muscular atrophy (SMA) is caused by deficiency of SMN (survival motor neuron), a ubiquitous and indispensable protein essential for biogenesis of snRNPs, key components of pre-mRNA processing. However, SMA's hallmark MN pathology, including neuromuscular junction (NMJ) disruption and sensory-motor circuitry impairment, remains unexplained. Toward this end, we used deep RNA sequencing (RNA-seq) to determine if there are any transcriptome changes in MNs and surrounding spinal cord glial cells (white matter, WM) microdissected from SMN-deficient SMA mouse model at presymptomatic postnatal day 1 (P1), before detectable MN pathology (P4-P5). The RNA-seq results, previously unavailable for SMA at any stage, revealed cell-specific selective mRNA dysregulations (~300 of 11,000 expressed genes in each, MN and WM), many of which are known to impair neurons. Remarkably, these dysregulations include complete skipping of agrin's Z exons, critical for NMJ maintenance, strong up-regulation of synapse pruning-promoting complement factor C1q, and down-regulation of Etv1/ER81, a transcription factor required for establishing sensory-motor circuitry. We propose that dysregulation of such specific MN synaptogenesis genes, compounded by many additional transcriptome abnormalities in MNs and WM, link SMN deficiency to SMA's signature pathology.

Effect of RGD-disintegrins on melanoma cell growth and metastasis: involvement of the actin cytoskeleton, FAK and c-Fos.

The effects and molecular mechanisms of RGD-disintegrins isolated from snake venoms on the growth and metastatic potential of B16F10-melanoma cells were investigated. Jarastatin (JT) from Bothrops jararaca is a ligand of alpha(5)beta(1), alpha(v)beta(3) and alpha(m)beta(2) integrins, flavoridin (FL) from Trimeresurus flavoridis binds preferentially to alpha(5)beta(1) and kistrin (KR) from Calloselasma rhodostoma is a selective ligand of alpha(v)beta(3). When injected simultaneously with melanoma cells in mice, the three disintegrins significantly reduced tumor lung colonization. On the other hand, JT and FL, but not KR, inhibited B16F10 cell growth in vitro. Interaction of JT or FL with melanoma cells induced actin cytoskeleton rearrangement, increasing actin polymerization and FAK phosphorylation. The effect of FL correlates with the decrease in the constitutively high nuclear content of c-Fos, whereas JT interfered with NF-kappaB translocation in melanoma cells. None of the disintegrins produced alterations in the nuclear Erk-2. The results provide further evidence to suggest RGD-disintegrins as potent anti-metastatic agents in vivo, and indicate that their interaction with alpha(5)beta(1) integrin interfere with integrin-couple signaling, down-regulating transcription factors and negatively modulating cell proliferation. These effects may contribute to inhibition of melanoma cell invasion in vivo.

Interaction of alpha9beta1 integrin with thrombospondin-1 promotes angiogenesis.

Thrombospondin-1 is a multifunctional protein interacting with several cell surface receptors including integrins. We found that it is a ligand for alpha9beta1 integrin, and has an integrin binding site within its N-terminal domain (NoC1). Interaction of thrombospondin-1 and its recombinant NoC1 domain with alpha9beta1 integrin was confirmed in ELISA and cell adhesion assays. Binding of NoC1 to cells expressing alpha9beta1 integrin activated signaling proteins such as Erk1/2 and paxillin. Blocking of this integrin by monoclonal antibody and the met-leu-asp-disintegrin inhibited dermal human microvascular endothelial cell proliferation and NoC1-induced migration of these cells. Immunohistochemical studies revealed that alpha9beta1 is expressed on microvascular endothelium in several organs including skin, lung, heart and brain. NoC1 induced neovascularization in an experimental quail chorioallantoic membrane system and Matrigel plug formation assay in mice. This proangiogenic activity of NoC1 in vivo was inhibited by alpha9beta1 inhibitors. In summary, our results revealed that alpha9beta1 integrin expressed on microvascular endothelial cells interacts with thrombospondin-1, and this interaction is involved in modulation of angiogenesis.