Recombinant proteins as promising antigens applied to the immunodiagnosis of Chagas disease: a scoping review.

Chagas disease (CD), caused by the protozoan Trypanosoma cruzi, is an important public health problem, occurring mainly in Latin America. The disease has a major social and economical effect, negatively impacting the life of the infected individuals, and bringing great costs to public health. An early and accurate diagnosis is essential for administration of early treatment. In addition, prognostic tests may aid disease management, decreasing hospitalization costs. However, the serological diagnostic scenario for CD still faces several challenges, making the development of new diagnostic kits a pressing matter. Facing this scenario, several researchers have expanded efforts in developing and testing new antigens, such as recombinant proteins and recombinant multiepitope proteins, with promising results. These recombinant antigens offer several advantages, such as improved sensitivity and specificity, in addition to facilitated scaling. Also, it has been possible to observe a rising number of studies using ELISA and point-of-care platforms, employing these antigens in the past few years. Among them, recombinant proteins were the most applied antigens, demonstrating great capacity to discriminate between positive and negative samples. Although fewer in number, recombinant multiepitope proteins also demonstrated an improved diagnostic performance. Indeed, a great number of studies employing these antigens showed sensitivity and specificity values above 90%, greatly impacting diagnostic accuracy. Nevertheless, despite the good results found, it is still possible to observe some bottlenecks in the development of new antigens, such as the scarcity of tests with sera from the acute phase and the variability of results in different geographic areas. In this sense, aiming to contribute to control and health programs, the continuous search for a more accurate serological diagnosis is essential, both for the acute and chronic phases of the disease.

Subjective sleep assessment in individuals with SYNGAP1-associated syndrome.

STUDY OBJECTIVES: Sleep disturbances are common in neurodevelopmental disorders (NDDs), affecting patients and caregivers' quality of life. SYNGAP1-associated syndrome, a rare NDD, is marked by intellectual disability, developmental delay, epilepsy, and sleep issues. However, research on sleep quality in these individuals is limited. This study aimed to evaluate genetic variants, epilepsy, and sleep patterns in SYNGAP1-associated syndrome patients and their caregivers. METHODS: An online survey was applied to 11 caregivers of individuals diagnosed with SYNGAP1-associated syndrome. Specific clinical inquiries were included, addressing childbirth, previous surgeries, and medication use. Inquiries about epilepsy included type of epilepsy, type and frequency of seizures, anti-seizure medications, and complementary non-pharmacological treatments. Children's Sleep Habits Questionnaire (CSHQ) was applied to assess the patients' sleep profile. Pittsburgh Sleep Quality Index (PSQI) was used to evaluate the sleep quality of caregivers. RESULTS: Genetic analysis showed heterozygous mutations in SYNGAP1, often leading to loss of function. Epilepsy was present in 82% of participants, with 77.8% having drug-resistant seizures. Using the Children's Sleep Habits Questionnaire (CSHQ), 81.8% of patients exhibited poor sleep habits, including bedtime resistance, anxiety, night awakenings, parasomnias, and daytime sleepiness. Caregivers also reported poor sleep quality according to the Pittsburgh Sleep Quality Index (PSQI). CONCLUSIONS: This study highlights the high prevalence of epilepsy and sleep problems in SYNGAP1-associated syndrome, impacting both patients and caregivers. Further research is crucial to understand the syndrome's effects on sleep disturbances, emphasizing the need for targeted interventions to improve sleep quality in individuals with rare genetic syndromes and their caregivers.

The use of peptides for immunodiagnosis of human Chagas disease.

Chagas disease, caused by the protozoa Trypanosoma cruzi, continues to be a serious public health problem in Latin America, worsened by the limitations in its detection. Given the importance of developing new diagnostic methods for this disease, the present review aimed to verify the number of publications dedicated to research on peptides that demonstrate their usefulness in serodiagnosis. To this end, a bibliographic survey was conducted on the PubMed platform using the keyword "peptide" or "epitope" combined with "Chagas disease" or "Trypanosoma cruzi"; "diagno*" or "serodiagnosis" or "immunodiagnosis", without period restriction. An increasing number of publications on studies employing peptides in ELISA and rapid tests assays was verified, which confirms the expansion of research in this field. It is possible to observe that many of the peptides tested so far originate from proteins widely used in the diagnosis of Chagas, and many of them are part of commercial tests developed. In this sense, as expected, promising results were obtained for several peptides when tested in ELISA, as many of them exhibited sensitivity and specificity values above 90%. Furthermore, some peptides have been tested in several studies, confirming their diagnostic potential. Despite the promising results observed, it is possible to emphasize the need for extensive testing of peptides, using different serological panels, in order to confirm their potential. The importance of producing an effective assay capable of detecting the clinical stages of the disease, as well as new immunogenic antigens that enable new serological diagnostic tools for Chagas disease, is evident.

Recombinant multiepitope proteins expressed in Escherichia coli cells and their potential for immunodiagnosis.

Recombinant multiepitope proteins (RMPs) are a promising alternative for application in diagnostic tests and, given their wide application in the most diverse diseases, this review article aims to survey the use of these antigens for diagnosis, as well as discuss the main points surrounding these antigens. RMPs usually consisting of linear, immunodominant, and phylogenetically conserved epitopes, has been applied in the experimental diagnosis of various human and animal diseases, such as leishmaniasis, brucellosis, cysticercosis, Chagas disease, hepatitis, leptospirosis, leprosy, filariasis, schistosomiasis, dengue, and COVID-19. The synthetic genes for these epitopes are joined to code a single RMP, either with spacers or fused, with different biochemical properties. The epitopes' high density within the RMPs contributes to a high degree of sensitivity and specificity. The RMPs can also sidestep the need for multiple peptide synthesis or multiple recombinant proteins, reducing costs and enhancing the standardization conditions for immunoassays. Methods such as bioinformatics and circular dichroism have been widely applied in the development of new RMPs, helping to guide their construction and better understand their structure. Several RMPs have been expressed, mainly using the Escherichia coli expression system, highlighting the importance of these cells in the biotechnological field. In fact, technological advances in this area, offering a wide range of different strains to be used, make these cells the most widely used expression platform. RMPs have been experimentally used to diagnose a broad range of illnesses in the laboratory, suggesting they could also be useful for accurate diagnoses commercially. On this point, the RMP method offers a tempting substitute for the production of promising antigens used to assemble commercial diagnostic kits.

A Review on the use of Synthetic and Recombinant Antigens for the Immunodiagnosis of Tegumentary Leishmaniasis.

Improving the diagnostic technology used to detect tegumentary leishmaniasis (TL) is essential in view of it being a widespread, often neglected tropical disease, with cases reported from the Southern United States to Northern Argentina. Recombinant proteins, recombinant multiepitope proteins, and synthetic peptides have been extensively researched and used in disease diagnosis. One of the benefits of applying these antigens is a measurable increase in sensitivity and specificity, which improves test accuracy. The present review aims to describe the use of these antigens and their diagnostic effectiveness. With that in mind, a bibliographic survey was conducted on the PudMed platform using the search terms "tegumentary leishmaniasis" AND "diagno", revealing that recombinant proteins have been described and evaluated for their value in TL diagnosis since the 1990s. However, there was a spike in the number of publications using all of the antigens between 2013 and 2022, confirming an expansion in research efforts to improve diagnosis. Moreover, all of the studies involving different antigens had promising results, including improved sensitivity and specificity. These data recognize the importance of doing research with new technologies focused on developing quick, more effective diagnostic kits as early diagnosis facilitates treatment.

Molecular methods for diagnosis of monkeypox: A mini-review.

BACKGROUND: Monkeypox is a global public health issue caused by the monkeypox virus (MPXV). As of October 28, 2022, a total of 77,115 laboratory-confirmed cases and 3,610 probable cases, including 36 deaths, were reported, with 9,070 cases reported in Brazil, the second most affected country. The need to develop national technologies for the rapid diagnosis of emerging diseases for mass testing of the population is evident, as observed in the SARS-CoV-2 pandemic. OBJECTIVE: With that in mind, this article provides an overview of current methods, techniques, and their applications in the molecular detection of monkeypox, focusing the search on real-time polymerase chain reaction (qPCR), polymerase chain reaction (PCR), and polymerase chain reaction-enzyme linked immunosorbent assay (PCR-ELISA). METHODS: The relevant documents or papers covered in this study were selected by a search in international bibliographic databases. The search terms used in the databases were aimed at summarizing existing knowledge on molecular diagnostic methods, such as monkeypox; MPX, MPXV, qPCR, PCR, PCR-ELISA, diagnosis and detection searched separately or together using the Boolean operator "AND" either in the title or abstract. The searches took place in September 2022, and the corresponding articles were selected between 2012 and 2022. RESULTS: We found 256 documents in total and twelve studies addressing the molecular diagnosis of monkeypox were classified as possible sources for this review. CONCLUSION: It is evident there is a pressing need to develop national technologies for rapid diagnosis of emerging diseases for mass testing of the population. It is also extremely important to have national detection kits with greater diagnostic capacity to assist in developing effective public policies in countries affected by this disease.

Dysfunctional Attitudes Toward Motherhood and Depressive Symptoms in Portuguese Pregnant Women During COVID-19 Pandemic: the Mediating Roles of Self-Compassion and Mindful Self-Care.

Objectives: There is evidence that the coronavirus disease 2019 (COVID-19) pandemic had a negative impact on the mental health of pregnant women, such as an increase in the prevalence of depression. Given the negative consequences that depressive symptoms can pose for pregnant women, it is crucial to understand how cognitive processes influence women's depressive symptoms during the COVID-19 pandemic among this specific vulnerable population. The present study aimed to examine the relationship between pregnant women's dysfunctional attitudes toward motherhood and their depressive symptoms, considering the mediating roles of self-compassion and mindful self-care. Method: A cross-sectional online survey was conducted in Portugal between October 2020 and April 2021. A sample of 244 pregnant women answered a set of questionnaires to assess dysfunctional attitudes toward motherhood, depressive symptoms, self-compassion, and mindful self-care. Results: More dysfunctional attitudes toward motherhood were associated with higher levels of depressive symptoms, and this relation was mediated through self-compassion and mindful self-care. Conclusions: These results highlight that self-compassion and mindful self-care are important psychological mechanisms and the importance of considering them in psychological interventions to prevent and/or treat depressive symptoms in pregnant women with dysfunctional attitudes toward motherhood during stressful events such as a pandemic.

Interest Groups as an Alternative for Medical Education in Sleep Medicine: Experience Report at a Brazilian Medical School.

Sleep medicine classes and teachings are usually deficient and insufficient during undergraduate medical education. In order to circumvent the educational deficits in sleep medicine, students at a Brazilian Medical School created a sleep medicine interest group-an academic organization for teaching purposes whose administration is carried out by the undergraduate students themselves. This study aims to describe the establishment of a sleep medicine interest group, as well as to evaluate the results of its first edition on the knowledge about sleep medicine among undergraduate medical students. Classes were taken biweekly and consisted of lectures by invited professors, presentation of clinical cases, and discussion with the students. By the end of the course, both attendees and non-attendees were invited to fill out a questionnaire including an objective assessment of knowledge (15 multiple choice questions). The questionnaire was filled out by 32 participants, of which 18 were attendees and 14 were non-attendees. The average result on the final exam was significantly higher among the attendees (6.1 ± 1.2) in comparison with non-attendees (4.9 ± 1.3-p = 0.015). The results demonstrate that an interest group proved to be feasible as a source of complementary information to undergraduate medical students and a valid alternative to circumvent the educational deficits.

Could melatonin have a potential adjuvant role in the treatment of the lasting anosmia associated with COVID-19? A review.

INTRODUCTION: Anosmia, the loss of the sense of smell, is usually associated with rhinopathies and has been reported as a common symptom of COVID-19. There is no specific drug to treat this condition, although some evidence suggests that melatonin could promote the recovery of olfactory sensory neurons. METHODS: We set out to perform a narrative review to synthesize the current evidence in this area in respect of our hypothesis that melatonin may be linked with anosmia and play a part in oxidative stress and the regulation of inflammation. The main electronic databases (MEDLINE/PubMed, Embase, and Cochrane) were searched. RESULTS: The search produced 26 articles related to our hypothesis. Some studies examined issues related to melatonin's effects and its use as adjuvant therapy for COVID-19. Despite some studies suggesting that melatonin may have potential in the treatment of COVID-19, to the best of our knowledge, there have been no trials that have used it to treat anosmia associated with the disease. Few articles identified proposed that melatonin might have an effect on olfactory cells. DISCUSSION: Further experimental and clinical research on the role of circadian melatonin in the olfactory system is warranted. This will provide evidence of the use of melatonin in the management of anosmia. A number of identified studies suggest that the imbalanced release of melatonin by the pineal gland associated with sleep disturbance may play a role in anosmia, although the specific pathway is not yet entirely clear. This may be a base for further research into the potential role of melatonin as adjuvant treatment of anosmia.

Be a Mom: Patterns of Program Usage and Acceptability Among Women With Low-Risk and High-Risk for Postpartum Depression.

Background: Be a Mom is a self-guided web-based intervention developed to prevent postpartum depression (PPD) symptoms and to promote maternal wellbeing, respectively among high and low-risk new mothers. This study aims to examine and compare (1) Be a Mom's patterns of usage and (2) Be a Mom's acceptability among women presenting high and low risk for PPD. Methods: The sample was composed by 800 women who were randomized to Be a Mom [542 presenting high-risk (Postpartum Depression Predictors Inventory-Revised ≥ 5.5) and 258 presenting low-risk for PPD]. Data regarding patterns of usage were collected through the Be a Mom website. Acceptability data were collected through a brief questionnaire. Results: 27.9% of high-risk and 36.3% of low-risk women completed the program. A higher proportion of participants in the low-risk group completed Be a Mom [ X ( 1 ) 2 = 5.29, p = 0.021] and completed more modules [t (723) = -3.01, p = 0.003]. No significant differences were found between the groups in number of logins, minutes spent on the program, exercises completed and number of times audios were played. a higher proportion of women in the high-risk group considered that participating in Be a Mom was too demanding [ X ( 1 ) 2 = 8.21, p = 0.004]. Conclusions: Despite the low rates of completion, Be a Mom appears to be an acceptable option for both women with high-risk and low-risk for PPD. Lack of time seems to be the main reason for non-completion, so it is important to develop briefer versions of the program and introduce engagement strategies that may increase completion rate.

Self-compassion and complete perinatal mental health in women at high risk for postpartum depression: The mediating role of emotion regulation difficulties.

OBJECTIVES: There is evidence suggesting that self-compassion is linked to key mechanisms in emotion regulation models of depression and anxiety. However, the majority of prior research has targeted community samples and overlooked the analysis of such mechanisms in relation to positive mental health. This study sought to examine the direct and indirect effects, via emotion regulation difficulties, of self-compassion on the complete mental health outcomes of women at high risk for postpartum depression (PPD). DESIGN: Cross-sectional, correlational study. METHODS: The sample for this study comprised 1053 postpartum women presenting high risk for PPD, as determined by the clinical cut-off score obtained through the Postpartum Depression Predictors Inventory-Revised. The assessment protocol further included self-report measures of self-compassion, depression and anxiety symptoms, flourishing and emotion regulation difficulties. Correlation analyses were conducted to ascertain associations between variables, and path analysis was performed to examine direct and indirect effects. RESULTS: Difficulties in emotion regulation mediated the links between self-compassion and negative (distress) and positive (flourishing) mental health outcomes. The overall mediation model explained 48% and 39% of variance for psychological distress and flourishing/positive mental health respectively. CONCLUSIONS: These findings support the applicability of an emotion regulation model of self-compassion to women identified as being at high risk of developing PPD. Self-compassion interventions directed at these women are likely to decrease their levels of emotion dysregulation and to improve their complete mental health outcomes.

Sleep in Older Adults and Its Possible Relations With COVID-19.

Since the beginning of the COVID-19 pandemic, older adults have been found to be a highly vulnerable group, with a higher prevalence of severe cases and negative outcomes. Research has focused on the reasons why older adults are at greater risk; Sleep-related factors have been suggested as one possible explanation for this. An individual's sleep pattern undergoes significant changes over the course of their life. In older adults a specific sleep profile can be observed, one characterized by advanced sleep timing, a morningness preference, longer sleep-onset latency, shorter overall sleep duration, increased sleep fragmentation, reduced slow-wave sleep and, increased wake time after sleep onset. Additionally, an increased prevalence of sleep disorders can be observed, such as obstructive sleep apnea and insomnia. Previous research has already linked sleep disorders (especially sleep apnea) with COVID-19, but few studies have focused specifically on the older population. We believe that the intrinsic sleep patterns of older adults, and the prevalence of sleep disorders in this population, may be important factors that could explain why they are at a greater risk of negative COVID-19 outcomes. In this review, we discuss the relationship between sleep and COVID-19 among older adults, focusing on three different aspects: (1) Sleep-related issues that might increase the likelihood of getting infected by SARS-COV-2; (2) Sleep disturbances that might increase the predisposition to worse COVID-19 prognosis and outcomes; and (3) COVID-19-related aspects affecting community-dwelling older adults, such as social isolation, quarantine, and home confinement, among others, that might impact sleep.

Temporal Association Between Respiratory Events and Reflux in Patients With Obstructive Sleep Apnea and Laryngopharyngeal Reflux.

STUDY OBJECTIVES: The aim of the current study was to test the hypothesis that there is a temporal correlation between reflux episodes and respiratory events in patients with laryngopharyngeal reflux and obstructive sleep apnea. METHODS: Adults with clinically diagnosed laryngopharyngeal reflux confirmed by two validated instruments (reflux symptom index ≥ 13 and reflux finding score ≥ 7) and obstructive sleep apnea (OSA) underwent full polysomnography with concomitant and synchronized multichannel intraluminal impedance-pH esophageal monitoring. The apnea-hypopnea and arousal indexes that occurred 15, 30, and 45 minutes before and after each reflux episode were recorded and compared to full-night apnea and hypopnea and arousal index. RESULTS: We studied 27 patients (14 males, age 51.7 ± 9.1 years, body mass index 32.4 ± 4.2 kg/m²) with laryngopharyngeal reflux (reflux symptom index 16 ± 2 and reflux finding score 12 ± 3) and OSA (apnea-hypopnea index = 32.3 ± 28.4 events/h). We evaluated 102 reflux episodes. Almost half of the reflux episodes occurred while awake (43.1%) and only five reflux episodes (4.9%) occurred during an obstructive respiratory event. The apnea and hypopnea and arousal indexes 15, 30, and 45 minutes before and after reflux episodes were lower than full-night apnea and hypopnea and arousal indexes, respectively. CONCLUSIONS: Among patients with well-established laryngopharyngeal reflux and OSA, there is no temporal association between reflux and obstructive respiratory events. Even though the data comprised a small sample size, it seems that a more complex mechanism is involved with these two highly prevalent diseases.

Comparison between flow cytometry and standard PCR in the evaluation of MRD in children with acute lymphoblastic leukemia treated with the GBTLI LLA - 2009 protocol.

Minimal residual disease (MRD) monitoring is of prognostic importance in childhood acute lymphoblastic leukemia (ALL). The detection of immunoglobulin and T-cell receptor gene rearrangements by real-time quantitative PCR (RT-PCR) is considered the gold standard for this evaluation. However, more accessible methods also show satisfactory performance. This study aimed to compare MRD analysis by four-color flow cytometry (FC) and qualitative standard PCR on days 35 and 78 of chemotherapy and to correlate these data with patients' clinical characteristics. Forty-two children with a recent diagnosis of ALL, admitted to a public hospital in Brazil for treatment in accordance with the Brazilian Childhood Cooperative Group for ALL Treatment (GBTLI LLA-2009), were included. Bone marrow samples collected at diagnosis and on days 35 and 78 of treatment were analyzed for the immunophenotypic characterization of blasts by FC and for the detection of clonal rearrangements by standard PCR. Paired analyses were performed in 61/68 (89.7%) follow-up samples, with a general agreement of 88.5%. Disagreements were resolved by RT-PCR, which evidenced one false-negative and four false-positive results in FC, as well as two false-negative results in PCR. Among the prognostic factors, a significant association was found only between T-cell lineage and MRD by standard PCR. These results show that FC and standard PCR produce similar results in MRD detection of childhood ALL and that both methodologies may be useful in the monitoring of disease treatment, especially in regions with limited financial resources.

Statistical control of the production of blood components by control charts of attribute to improve quality characteristics and to comply with current specifications.

Statistical process control (SPC) is closely related to good quality control practices in the manufacturing process. One of the primary goals is to detect unnatural patterns, allowing the production service to control the conformity of the blood components produced. Despite being recommended by national and international standards, its exercise is not uniform, and sometimes the methodology used is misinterpreted as SPC. When the input data has a Gaussian distribution, control charts for variables are proposed. However, when the data distribution is not normal, control charts for attributes are suggested. This article presents and discusses four statistical procedures for the control of attributes using p-, np-, u-, and c-charts. An empirical demonstration shows these models are reliable for in routine use in the Blood Establishment quality control, as also suggests the use when the control charts for variables are inapplicable.

Statistical methods to the control of the production of blood components: principles and control charts for variables.

General quality control good practices require the control of the production of blood components using statistical techniques, such as mandatory by the European Commission Directives and the American Association of Blood Banks standards. Sometimes, the control procedure is exclusively in favor of the compliance verification with specifications per individual component or to compute the number of defective parts usually on a monthly basis. However, this is a critical restriction to detect unnatural patterns such as to guarantee that the production has a non-significance chance to manufacturing nonconforming components. Therefore, a crucial issue in Blood Establishments is the application of a reliable statistical process control methodology to assure products reliable and consistent to specifications. Statistical principles and control charts for variables are reviewed, discussed and recommended, based on current good practices. The empirical data demonstrate the consistency of these models on blood establishment routine. A flowchart to select the type of control chart is suggested.