Sepsis expands a CD39+ plasmablast population that promotes immunosuppression via adenosine-mediated inhibition of macrophage antimicrobial activity.

Sepsis results in elevated adenosine in circulation. Extracellular adenosine triggers immunosuppressive signaling via the A2a receptor (A2aR). Sepsis survivors develop persistent immunosuppression with increased risk of recurrent infections. We utilized the cecal ligation and puncture (CLP) model of sepsis and subsequent infection to assess the role of adenosine in post-sepsis immune suppression. A2aR-deficient mice showed improved resistance to post-sepsis infections. Sepsis expanded a subset of CD39hi B cells and elevated extracellular adenosine, which was absent in mice lacking CD39-expressing B cells. Sepsis-surviving B cell-deficient mice were more resistant to secondary infections. Mechanistically, metabolic reprogramming of septic B cells increased production of ATP, which was converted into adenosine by CD39 on plasmablasts. Adenosine signaling via A2aR impaired macrophage bactericidal activity and enhanced interleukin-10 production. Septic individuals exhibited expanded CD39hi plasmablasts and adenosine accumulation. Our study reveals CD39hi plasmablasts and adenosine as important drivers of sepsis-induced immunosuppression with relevance in human disease.

The impact and the challenges of implementing a faculty development program on health professions education in a Brazilian Medical School: a case study with mixed methods.

PURPOSE: Faculty development in health professions education is still challenging in developing countries like Brazil. Work overload and the lack of financial support hinder faculty members' participation. Ribeirão Preto Medical School founded its Center for Faculty Development in 2016. Since then, an essential skills module (ESMo) on health professions education (HPE) has been offered regularly to faculty members and preceptors of seven undergraduate programs. This case study aims to evaluate the impact of this Essential Skills Module on the educational practices of participants two years after attending the module and the challenges faced during the process. METHOD: The study used a mixed-method approach with a description of the demographic and professional profile data of the ESMo participants. Immediate post-ESMo perceptions (satisfaction and learning) of the participants were determined with structured instruments. Two years later, a semi-structured interview was conducted and recorded to determine the long-term effects (application of learning and behavior changing as an educator). NVIVO® software was used to store and systematize the thematic discourse analysis with a socio-constructivist theoretical framework interpretation. RESULTS: One hundred forty-six participants were included: 86 (59%) tenured faculty members, 49 (33,5%) clinical preceptors, and 11 (7,5%) invited teachers. Most were female (66%), and 56% had teaching experience shorter than ten years. 52 (69%) out of 75 eligible participants were interviewed. The immediate reaction to participating in the module was quite positive and 80% have already implemented an educational intervention in their daily activities. Discourses thematic analysis showed five emerging themes appearing in different frequencies: Changes in teaching activities (98%); Lack of previous pedagogical training (92.3%); Commitment and enthusiasm towards teaching (46.15%); Overlapping functions inside the institution (34.6%) and Challenges for student assessment (23%). CONCLUSION: This first in-depth evaluation of the long-term effects of a faculty development intervention in a Brazilian Health Profession Education school showed that participation positively changed participants' teaching & learning practices. These interventions consistently fostered a community of practice and valued faculty development processes in local and national scenarios.

Exposure to high endotoxin concentration increases wheezing prevalence among laboratory animal workers: a cross-sectional study.

BACKGROUND: Endotoxin from Gram-negative bacteria are found in different concentrations in dust and on the ground of laboratories dealing with small animals and animal houses. METHODS: Cross-sectional study performed in workplaces of two universities. Dust samples were collected from laboratories and animal facilities housing rats, mice, guinea pigs, rabbits or hamsters and analyzed by the "Limulus amebocyte lysate" (LAL) method. We also sampled workplaces without animals. The concentrations of endotoxin detected in the workplaces were tested for association with wheezing in the last 12 months, asthma defined by self-reported diagnosis and asthma confirmed by bronchial hyperresponsiveness (BHR) to mannitol. RESULTS: Dust samples were obtained at 145 workplaces, 92 with exposure to animals and 53 with no exposure. Exposed group comprised 412 subjects and non-exposed group comprised 339 subjects. Animal-exposed workplaces had higher concentrations of endotoxin, median of 34.2 endotoxin units (EU) per mg of dust (interquartile range, 12.6-65.4), as compared to the non-exposed group, median of 10.2 EU/mg of dust (interquartile range, 2.6-22.2) (p < 0.001). The high concentration of endotoxin (above whole sample median, 20.4 EU/mg) was associated with increased wheezing prevalence (p < 0.001), i.e., 61 % of workers exposed to high endotoxin concentration reported wheezing in the last 12 months compared to 29 % of workers exposed to low endotoxin concentration. The concentration of endotoxin was not associated with asthma report or with BHR confirmed asthma. CONCLUSION: Exposure to endotoxin is associated with a higher prevalence of wheezing, but not with asthma as defined by the mannitol bronchial challenge test or by self-reported asthma. Preventive measures are necessary for these workers.

A Functional Bread Fermented with Saccharomyces cerevisiae UFMG A-905 Prevents Allergic Asthma in Mice.

Background: The administration of probiotics has been shown to be beneficial in asthma. The administration of Saccharomyces cerevisiae UFMG A-905 prevented asthma development. Traditionally, probiotics are administered using dairy-based matrices, but other vehicles (e.g., fruit juices, biscuits, candies, and breads) can be used. Objectives: This study aimed to assess the effect of bread fermented with S. cerevisiae UFMG A-905 in asthma prevention. Methods: Three breads were produced: fermented with commercial yeast, fermented with S. cerevisiae UFMG A-905, and fermented with S. cerevisiae UFMG A-905 with the addition of alginate microcapsules containing live S. cerevisiae UFMG A-905. Characterization of the microbial composition of the breads was performed. Male Balb/c mice were sensitized and challenged with ovalbumin. Breads were administered 10 d before the first sensitization and during sensitization and challenge protocol. Yeast fecal count, in vivo airway hyperresponsiveness, and airway and lung inflammation were assessed. Results: In UFMG A-905 bread, there was an increase in yeast number and a decrease in total and lactic acid bacteria. Animals that received S. cerevisiae UFMG A-905 fermented bread with microcapsules had a significant increase in yeast recovery from feces. S. cerevisiae UFMG A-905-fermented breads partially reduced airway inflammation, decreasing eosinophils and IL5 and IL13 concentrations. When adding microcapsules, the bread also diminished airway hyperresponsiveness and increased IL17A concentrations. Conclusions: S. cerevisiae UFMG A-905 was able to generate long-fermentation breads. Microcapsules were a safe and viable way to inoculate the live yeast into food. The administration of breads fermented with S. cerevisiae UFMG A-905 prevented asthma-like characteristics, being more pronounced when the breads contained microcapsules with live yeast.

Chloroquine use improves dengue-related symptoms.

Dengue is the most important arboviral disease in the world. As chloroquine, an antimalarial agent, has shown some antiviral effects, this study evaluated its effect in patients with dengue. A randomised, double-blind study was performed by administering chloroquine or placebo for three days to 129 patients with dengue-related symptoms. Of these patients, 37 were confirmed as having dengue and completed the study; in total, 19 dengue patients received chloroquine and 18 received placebo. There was no significant difference in the duration of the disease or the degree and days of fever. However, 12 patients (63%) with confirmed dengue reported a substantial decrease in pain intensity and a great improvement in their ability to perform daily activities (p = 0.0004) while on the medication and the symptoms returned immediately after these patients stopped taking the medication. The same effect was not observed in patients with diseases other than dengue. Therefore, this study shows that patients with dengue treated with chloroquine had an improvement in their quality of life and were able to resume their daily activities. However, as chloroquine did not alter the duration of the disease or the intensity and days of fever, further studies are necessary to confirm the clinical effects and to assess the side effects of chloroquine in dengue patients.

Antibody response dynamics to CoronaVac vaccine and booster immunization in adults and the elderly: A long-term, longitudinal prospective study.

Background: The long-term humoral immune response after vaccination varies between vaccines and is dependent on the accuracy of the antibody test. A better understanding of the vaccine immune response may help to define vaccination strategies against coronavirus disease 2019 (COVID-19). Objective: To investigate the long-term immunological response to CoronaVac vaccine and determinants of breakthrough COVID-19 infection. Methods: A long-term, prospective cohort study involving vaccinated adult and elderly subjects was conducted to investigate the presence of anti-RBD-specific immunoglobulin (Ig)G, anti-nucleocapsid IgG and anti-spike trimeric protein IgG. Antibody level dynamics and risk factors associated with breakthrough COVID-19 infection were investigated. Results: In total, 3902 participants were included in this study. Vaccination with two doses of CoronaVac and a booster dose increased the levels of anti-RBD-specific IgG, anti-nucleocapsid IgG and anti-spike trimeric IgG significantly. In adults, anti-nucleocapsid IgG and anti-spike trimeric IgG levels decreased significantly 7 months after the second dose. In adults and the elderly, the levels of anti-spike trimeric IgG and anti-RBD IgG decreased significantly 4 and 6 months after the booster dose, respectively. Previous exposure to severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) and anti-spike trimeric IgG titres was independently associated with a lower probability of post-vaccination infection. Conclusions: A significant increase in antibody levels was found after two doses of CoronaVac and a booster dose. Antibody titres declined significantly 7 months post-vaccination in participants who did not receive a booster dose. Higher levels of antibodies and previous SARS-CoV-2 infection were associated with protection against breakthrough COVID-19.

Impact of implementing an exclusively dedicated respiratory isolation room in a Brazilian tertiary emergency department.

BACKGROUND: Occupational risk due to airborne disease challenges healthcare institutions. Environmental measures are effective but their cost-effectiveness is still debatable and most of the capacity planning is based on occupational rates. Better indices to plan and evaluate capacity are needed. GOAL: To evaluate the impact of installing an exclusively dedicated respiratory isolation room (EDRIR) in a tertiary emergency department (ED) determined by a time-to-reach-facility method. METHODS: A group of patients in need of respiratory isolation were first identified--group I (2004; 29 patients; 44.1±3.4 years) and the occupational rate and time intervals (arrival to diagnosis, diagnosis to respiratory isolation indication and indication to effective isolation) were determined and it was estimated that adding an EDRIR would have a significant impact over the time to isolation. After implementing the EDRIR, a second group of patients was gathered in the same period of the year--group II (2007; 50 patients; 43.4±1.8 years) and demographic and functional parameters were recorded to evaluate time to isolation. Cox proportional hazard models adjusted for age, gender and inhospital respiratory isolation room availability were obtained. RESULTS: Implementing an EDRIR decreased the time from arrival to indication of respiratory isolation (27.5±9.3 × 3.7±2.0; p=0.0180) and from indication to effective respiratory isolation (13.3±3.0 × 2.94±1.06; p=0.003) but not the respiratory isolation duration and total hospital stay. The impact on crude isolation rates was very significant (8.9 × 75.4/100.000 patients; p<0.001). The HR for effective respiratory isolation was 26.8 (95% CI 7.42 to 96.9) p<0.001 greater for 2007. CONCLUSION: Implementing an EDRIR in a tertiary ED significantly reduced the time to respiratory isolation.

Advantages and pitfalls of the polymerase chain reaction in the diagnosis of esophageal ulcers in AIDS patients.

HIV-1-infected patients frequently have opportunistic esophageal infections which, when associated with severe immunodeficiency, can be attributed to unusual pathogens. The clinical presentation of several esophageal diseases is similar and the best method for a specific diagnosis of these patients has not been well defined. To evaluate the role of the polymerase chain reaction (PCR) in the etiologic definition of esophageal ulcers in HIV-1-infected patients, 96 esophageal biopsies from 79 HIV-1-infected patients were processed by PCR using specific primers for cytomegalovirus (CMV), herpes virus (HSV), human papilloma virus (HPV), HIV-1, Mycobacterium tuberculosis, Mycobacterium avium, Mycobacterium intracellulare, Treponema pallidum, and Haemophilus ducreyi. The PCR results were compared to the histopathologic results. Seventy-nine patients were studied (mean age: 34 years; 62% men; median CD4 + T cell = 103.59 cells/microl (range 1-795.2 cells/microl). The most common endoscopic findings were as follows: esophageal candidiasis (37.1%), esophageal ulcers (24.7%), esophagitis (11.2%), and lugol-negative areas (10.1%). The histopathologic findings in the esophageal ulcers (22 biopsies) were non-specific inflammation (31.8%), HSV (36.4%), Candida (13.6%), CMV (13.6%), or HPV disease (4.5%). In the esophageal ulcer biopsies, the PCR results were negative in 27.6% of cases, and positive for HIV (65.5%), CMV (31%), HPV (20.7%), HSV (10.3%), and H. ducreyi (6.9%). The histopathologic examination did not identify a pathogen or identified only Candida in 15 biopsies of esophageal ulcers. PCR was positive in ten (66.7%) and negative in five (33.3%) of these biopsies (idiopathic ulcers). PCR detected: HIV (53.3%), CMV (20%), HPV (13.3%), and H. ducreyi (6,7%). PCR detected more etiologic agents in esophageal ulcers than histopathology and was able to detect unusual pathogens. On the other hand, sometimes more than one pathogen was detected in the esophageal ulcers, making it difficult to reach an accurate diagnosis. This finding indicates the need for more studies to evaluate the benefit of this method in the routine evaluation of esophageal ulcer biopsies in HIV-1-infected patients.

Aqueous extracts from Uncaria tomentosa (Willd. ex Schult.) DC. reduce bronchial hyperresponsiveness and inflammation in a murine model of asthma.

ETHNOPHARMACOLOGICAL RELEVANCE: Uncaria tomentosa (Willd. Ex Schult) DC is used by indigenous tribes in the Amazonian region of Central and South America to treat inflammation, allergies and asthma. The therapeutic properties of U. tomentosa have been attributed to the presence of tetracyclic and pentacyclic oxindole alkaloids and to phenolic acids. AIMS OF THE STUDY: To characterize aqueous bark extracts (ABE) and aqueous leaf extracts (ALE) of U. tomentosa and to compare their anti-inflammatory effects. MATERIALS AND METHODS: Constituents of the extracts were identified by ultra performance liquid chromatography-mass spectrometry. Anti-inflammatory activities were assessed in vitro by exposing lipopolysaccharide-stimulated macrophage cells (RAW264.7-Luc) to ABE, ALE and standard mitraphylline. In vivo assays were performed using a murine model of ovalbumin (OVA)-induced asthma. OVA-sensitized animals were treated with ABE or ALE while controls received dexamethasone or saline solution. Bronchial hyperresponsiveness, production of Th1 and Th2 cytokines, total and differential counts of inflammatory cells in the bronchoalveolar lavage (BAL) and lung tissue were determined. RESULTS: Mitraphylline, isomitraphylline, chlorogenic acid and quinic acid were detected in both extracts, while isorhyncophylline and rutin were detected only in ALE. ABE, ALE and mitraphylline inhibited the transcription of nuclear factor kappa-B in cell cultures, ALE and mitraphylline reduced the production of interleukin (IL)-6, and mitraphylline reduced production of tumor necrosis factor-alpha. Treatment with ABE and ALE at 50 and 200 mg kg-1, respectively, reduced respiratory elastance and tissue damping and elastance. ABE and ALE reduced the number of eosinophils in BAL, while ALE at 200 mg kg-1 reduced the levels of IL-4 and IL-5 in the lung homogenate. Peribronchial inflammation was significantly reduced by treatment with ABE and ALE at 50 and 100 mg kg-1 respectively. CONCLUSION: The results clarify for the first time the anti-inflammatory activity of U. tomentosa in a murine model of asthma. Although ABE and ALE exhibited distinct chemical compositions, both extracts inhibited the production of pro-inflammatory cytokines in vitro. In vivo assays revealed that ABE was more effective in treating asthmatic inflammation while ALE was more successful in controlling respiratory mechanics. Both extracts may have promising applications in the phytotherapy of allergic asthma.

Development and validation of an asthma knowledge questionnaire for use in Brazil.

OBJECTIVE: To develop and validate an asthma knowledge questionnaire for use in adult asthma patients in Brazil. METHODS: A 34-item self-report questionnaire was constructed and administered to adult asthma patients and adult controls. The maximum total score was 34. RESULTS: The questionnaire was shown to be discriminatory, with good reliability and reproducibility. The mean score for asthma patients and controls was, respectively, 21.47 +/- 4.11 (range: 9-31) and 17.27 +/- 5.11 (range: 7-28; p < 0.001). The Kaiser-Meyer-Olkin measure of sampling adequacy was 0.53, and the Bartlett's test of sphericity demonstrated a satisfactory suitability of the data to factor analysis (p < 0.001). There was no significant difference between the total scores obtained in the first and in the second application of the questionnaire within a two-week interval (p = 0.43). The internal consistency reliability (KR-20 coefficient) was 0.69. CONCLUSIONS: This study has validated an asthma knowledge questionnaire for use in Brazil.

Knowledge of and technique for using inhalation devices among asthma patients and COPD patients.

OBJECTIVE: To evaluate knowledge of and techniques for using prescribed inhalation devices among patients with asthma or COPD treated at a tertiary teaching hospital. METHODS: Patients were assessed after medical visits, and their physicians were blinded to this fact. Patients were asked to demonstrate their inhaler technique and were then interviewed regarding their knowledge of inhalation devices, control of the disease and instructions received during medical visits. RESULTS: We included 120 volunteers: 60 with asthma and 60 with COPD. All of the asthma patients and 98.3% of the COPD patients claimed to know how to use inhaled medications. In the sample as a whole, 113 patients (94.2%) committed at least one error when using the inhalation device. Patients committed more errors when using metered-dose inhalers than when using the dry-powder inhalers Aerolizer (p < 0.001) or Pulvinal (p < 0.001), as well as committing more errors when using the Aerolizer inhaler than when using the Pulvinal inhaler (p < 0.05). Using the metered-dose, Pulvinal and Aerolizer inhalers, the COPD group patients committed more errors than did the asthma group patients (p = 0.0023, p = 0.0065 and p = 0.012, respectively). CONCLUSIONS: Although the majority of the patients claimed to know how to use inhalation devices, the fact that 94.2% committed at least one error shows that their technique was inappropriate and reveals a discrepancy between understanding and practice. Therefore, it is not sufficient to ask patients whether they know how to use inhalation devices. Practical measures should be taken in order to minimize errors and optimize treatment.

Chloroquine use improves dengue-related symptoms

Dengue is the most important arboviral disease in the world. As chloroquine, an antimalarial agent, has shown some antiviral effects, this study evaluated its effect in patients with dengue. A randomised, double-blind study was performed by administering chloroquine or placebo for three days to 129 patients with dengue-related symptoms. Of these patients, 37 were confirmed as having dengue and completed the study; in total, 19 dengue patients received chloroquine and 18 received placebo. There was no significant difference in the duration of the disease or the degree and days of fever. However, 12 patients (63%) with confirmed dengue reported a substantial decrease in pain intensity and a great improvement in their ability to perform daily activities (p = 0.0004) while on the medication and the symptoms returned immediately after these patients stopped taking the medication. The same effect was not observed in patients with diseases other than dengue. Therefore, this study shows that patients with dengue treated with chloroquine had an improvement in their quality of life and were able to resume their daily activities. However, as chloroquine did not alter the duration of the disease or the intensity and days of fever, further studies are necessary to confirm the clinical effects and to assess the side effects of chloroquine in dengue patients.

Development and validation of an asthma knowledge questionnaire for use in Brazil / Desenvolvimento e validação de um questionário de conhecimento em asma para uso no Brasil

OBJECTIVE: To develop and validate an asthma knowledge questionnaire for use in adult asthma patients in Brazil. METHODS: A 34-item self-report questionnaire was constructed and administered to adult asthma patients and adult controls. The maximum total score was 34. RESULTS: The questionnaire was shown to be discriminatory, with good reliability and reproducibility. The mean score for asthma patients and controls was, respectively, 21.47 ± 4.11 (range: 9-31) and 17.27 ± 5.11 (range: 7-28; p < 0.001). The Kaiser-Meyer-Olkin measure of sampling adequacy was 0.53, and the Bartlett's test of sphericity demonstrated a satisfactory suitability of the data to factor analysis (p < 0.001). There was no significant difference between the total scores obtained in the first and in the second application of the questionnaire within a two-week interval (p = 0.43). The internal consistency reliability (KR-20 coefficient) was 0.69. CONCLUSIONS: This study has validated an asthma knowledge questionnaire for use in Brazil.

OBJETIVO: Desenvolver e validar um questionário de conhecimento em asma para pacientes adultos asmáticos no Brasil. MÉTODOS: Um questionário autoaplicável com 34 itens foi desenvolvido e aplicado em asmáticos e controles adultos. A pontuação total máxima era 34. RESULTADOS: O questionário mostrou-se discriminante, com boa confiabilidade e reprodutibilidade. O escore médio para os asmáticos e controles foi, respectivamente, 21,47 ± 4,11 (variação: 9-31) e 17,27 ± 5,11 (variação: 7-28; p < 0,001). O teste de Kaiser-Meyer-Olkin revelou uma medida de adequação de 0,53, e o teste de esfericidade de Bartlett demonstrou uma adequação satisfatória dos dados para a análise fatorial (p < 0,001). Não houve diferença significativa entre os escores totais obtidos na primeira e na segunda aplicação do questionário, com um intervalo de duas semanas (p = 0,43). O coeficiente de consistência interna (coeficiente KR-20) foi 0,69. CONCLUSÕES: Este estudo validou um questionário de educação em asma para uso no Brasil.

Técnica e compreensão do uso dos dispositivos inalatórios em pacientes com asma ou DPOC / Knowledge of and technique for using inhalation devices among asthma patients and COPD patients

OBJETIVO: Avaliar a compreensão e a técnica de uso dos dispositivos inalatórios prescritos de pacientes com asma ou DPOC atendidos em um hospital-escola terciário. MÉTODOS: Os pacientes foram avaliados na sala de pós-consulta sem que o médico soubesse do estudo. Foi solicitado que o paciente demonstrasse como utiliza a medicação inalatória e, em seguida, esse foi entrevistado sobre a compreensão dos dispositivos inalatórios, controle da doença e orientações recebidas durante as consultas. RESULTADOS: Foram avaliados 120 voluntários: 60 asmáticos e 60 com DPOC. Todos os asmáticos e 98,3 por cento do grupo DPOC relataram saber utilizar os medicamentos inalatórios. Na amostra como um todo, 113 pacientes (94,2 por cento) cometeram pelo menos um erro ao utilizar o dispositivo inalatório. Os pacientes cometeram mais erros ao utilizar aerossol dosimetrado do que ao utilizar os inaladores de pó seco Aerolizer® (p < 0,001) ou Pulvinal® (p < 0,001), assim como mais erros ao utilizar Aerolizer® do que ao utilizar Pulvinal® (p < 0,05). O grupo DPOC cometeu significativamente mais erros que o grupo asma ao utilizar o aerossol dosimetrado (p = 0,0023), Pulvinal® (p = 0,0065) e Aerolizer® (p = 0,012). CONCLUSÕES: Embora a maioria dos pacientes relatasse saber a técnica adequada de utilização dos dispositivos inalatórios, 94,2 por cento cometeu pelo menos um erro na utilização dos dispositivos, demonstrando técnica insatisfatória e discrepância entre a compreensão e a prática. Portanto, apenas questionar os pacientes sobre o uso dos dispositivos inalatórios não é suficiente. Medidas práticas devem ser tomadas a fim de diminuir os erros e otimizar a terapêutica.

OBJECTIVE: To evaluate knowledge of and techniques for using prescribed inhalation devices among patients with asthma or COPD treated at a tertiary teaching hospital. METHODS: Patients were assessed after medical visits, and their physicians were blinded to this fact. Patients were asked to demonstrate their inhaler technique and were then interviewed regarding their knowledge of inhalation devices, control of the disease and instructions received during medical visits. RESULTS: We included 120 volunteers: 60 with asthma and 60 with COPD. All of the asthma patients and 98.3 percent of the COPD patients claimed to know how to use inhaled medications. In the sample as a whole, 113 patients (94.2 percent) committed at least one error when using the inhalation device. Patients committed more errors when using metered-dose inhalers than when using the dry-powder inhalers Aerolizer® (p < 0.001) or Pulvinal® (p < 0.001), as well as committing more errors when using the Aerolizer® inhaler than when using the Pulvinal® inhaler (p < 0.05). Using the metered-dose, Pulvinal® and Aerolizer® inhalers, the COPD group patients committed more errors than did the asthma group patients (p = 0.0023, p = 0.0065 and p = 0.012, respectively). CONCLUSIONS: Although the majority of the patients claimed to know how to use inhalation devices, the fact that 94.2 percent committed at least one error shows that their technique was inappropriate and reveals a discrepancy between understanding and practice. Therefore, it is not sufficient to ask patients whether they know how to use inhalation devices. Practical measures should be taken in order to minimize errors and optimize treatment.