Rapid Classification of Serum from Patients with Paracoccidioidomycosis Using Infrared Spectroscopy, Univariate Statistics, and Linear Discriminant Analysis (LDA).

Paracoccidioidomycosis (PCM) is a systemic mycosis that is diagnosed by visualizing the fungus in clinical samples or by other methods, like serological techniques. However, all PCM diagnostic methods have limitations. The aim of this study was to develop a diagnostic tool for PCM based on Fourier transform infrared (FTIR) spectroscopy. A total of 224 serum samples were included: 132 from PCM patients and 92 constituting the control group (50 from healthy blood donors and 42 from patients with other systemic mycoses). Samples were analyzed by attenuated total reflection (ATR) and a t-test was performed to find differences in the spectra of the two groups. The wavenumbers that had p < 0.05 had their diagnostic potential evaluated using receiver operating characteristic (ROC) curves. The spectral region with the lowest p value was used for variable selection through principal component analysis (PCA). The selected variables were used in a linear discriminant analysis (LDA). In univariate analysis, the ROC curves with the best performance were obtained in the region 1551-1095 cm-1. The wavenumber that had the highest AUC value was 1264 cm-1, achieving a sensitivity of 97.73%, specificity of 76.01%, and accuracy of 94.22%. The total separation of groups was obtained in the PCA performed with a spectral range of 1551-1095 cm-1. LDA performed with the eight wavenumbers with the greatest weight from the group discrimination in the PCA obtained 100% accuracy. The methodology proposed here is simple, fast, and highly accurate, proving its potential to be applied in the diagnosis of PCM. The proposed method is more accurate than the currently known diagnostic methods, which is particularly relevant for a neglected tropical mycosis such as paracoccidioidomycosis.

Using infrared spectroscopy of serum and chemometrics for diagnosis of paracoccidioidomycosis.

Paracoccidioidomycosis (PCM) is a systemic granulomatous mycosis endemic to Latin America, whose etiologic agents are fungi of the genus Paracoccidioides. PCM is usually diagnosed by microscopic observation of the fungus in biological samples, combined or not with other techniques such as serological methods. However, all currently used diagnostic methods have limitations. The objective of this study was to develop a method based on Fourier transform infrared spectroscopy (FTIR) and chemometric analysis for PCM diagnosis. We included 224 serum samples: 132 PCM sera, 24 aspergillosis sera, 10 cryptococcosis sera, 8 histoplasmosis sera, and 50 sera from healthy blood donors. Samples were analyzed by attenuated total reflection (ATR), and chemometric analyses including exploratory analysis through principal component analysis (PCA) and a classification method (PCM and non-PCM) through orthogonal partial least squares discriminant analysis (OPLS-DA). The spectra were similar, with the main bands up to approximately 1652 cm-1 and 1543 cm-1 (amide I and amide II bands). This same region was mainly responsible for the partial separation of the samples in PCA. The OPLS-DA model correctly classified all serum samples with only one latent variable, with a determination coefficient (R²) higher than 0.999 for both the calibration set and prediction set. Sensitivity and specificity were 100% for both sets, showing better performance than the reference diagnostic methods. Therefore, the use of FTIR/ATR together with OPLS-DA modeling proved to be a promising method for PCM diagnosis.

Transitions in frailty state 12 months after kidney transplantation: a prospective cohort study.

BACKGROUND: Frailty is associated with several unfavorable outcomes after kidney transplantation (KTx). However, limited information is available regarding the transitions in frailty state and its components after KTx. This study aimed to evaluate the transitions in physical frailty phenotype and its components over a period of 12 months after KTx. METHODS: In this prospective single-center cohort study, we measured physical frailty phenotype (PFP) and its components at the time of admission for KTx and 12 months after KTx. The evaluation includes five components: weakness (grip strength analysed by sex and body mass index quartiles), physical activity (kcals/week based on the Minnesota Leisure Time Physical Activity questionnaire), exhaustion (self-report using the Center for Epidemiological Studies Depression Scale), gait speed (time taken to walk 15 feet based on sex and height-specific cutoff), and unintentional weight loss (self-report of unintentional weight loss > 10 lbs in the last year). The exhaustion and physical activity components are validated in the Brazilian population. Each component is scored as 0 or 1 according to its presence or absence, and a PFP score of 3-5 defines frailty, 2 is intermediate, and 0-1 is rated as non-frail. We used the McNemar and Wilcoxon test to compare physical frailty phenotype and its components between the two periods. RESULTS: Among 87 patients included in the study, 16.1% were classified as frail, 20.7% as intermediately frail, and 63.2% as non-frail. Sixty-four patients were included in the analysis to evaluate transitions in frailty. At the time of admission for KTx, 15.6% of patients were defined as frail compared to 4.7% of patients at 12 months after KTx (p = 0.023). Among the physical frailty phenotype components, the proportion of patients who scored in the weight loss category 12 months after KTx was significantly lower than that at the time of KTx (6.3% vs 34.4%, p < 0.001). CONCLUSIONS: There was a 69.9% reduction in the prevalence of frail patients at the end of the 12-month follow-up after KTx. Among the components of frailty, weight loss showed a significant improvement.

Frailty predicts surgical complications after kidney transplantation. A propensity score matched study.

BACKGROUND AND OBJECTIVE: Surgical complications after kidney transplantation can lead to catastrophic outcomes. Frailty has been associated with important kidney transplantation outcomes; however, there are no studies assessing this measure of physiological reserve as a specific predictor of surgical complications in this population. Such an assessment was, therefore, the objective of the present study. METHODS: A total of 87 individuals aged ≥ 18 years who underwent kidney transplantation between March 2017 and March 2018 were included. At the time of admission for kidney transplantation, demographic, clinical, and kidney transplantation data were collected, and the frailty score was calculated according to Fried et al., which comprises five components: shrinking, weakness, exhaustion, low activity, and slowed walking speed. Urological, vascular, and general surgical complications were assessed three months later, or until graft loss or death. The propensity score was used to achieve a better homogeneity of the sample, and new analyses were performed in this new, balanced sample. RESULTS: Of the 87 individuals included, 30 (34.5%) had surgical complications. After propensity score matching, the risk of surgical complications was significantly higher among the frail individuals (RR 2.14; 95% CI 1.01-4.54; p = 0.035); specifically, the risk of noninfectious surgical complications was significantly higher among these individuals (RR 2.50; 95% CI 1.11-5.62; p = 0.017). CONCLUSION: The results showed that individuals with some degree of frailty before kidney transplantation were more subject to surgical complications. The calculation of the frailty score for transplant candidates and the implementations of measures to increase the physiological reserve of these patients at the time of kidney transplantation may possibly reduce the occurrence of surgical complications.

Cefazolin-gentamicin versus taurolidine-citrate for the prevention of infection in tunneled central catheters in hemodialysis patients: A quasi-experimental trial.

BACKGROUND: Catheter-related bloodstream infection (CR-BSI) is one of various complications related to hemodialysis (HD). As a result of the high rate of infection, the use of lock solutions for the prevention of CR-BSI has been studied. However, adverse effects of lock solution, such as increased emergence of strains resistant to antibiotics, which is an important concern, need to be investigated further. The aim of this study was to compare the efficacy of lock solution using a combination of cefazolin and gentamicin versus taurolidine and citrate in reducing CR-BSI in patients undergoing HD and to identify any adverse effects. METHODS: A prospective observational study was performed at two dialysis centers. Patients using new tunneled central venous catheters (CVC) for HD were included. Patients with a tunneled CVC were assigned to receive either antibiotic lock solution (group 1: gentamicin 7mg/ml+cefazolin 12mg/ml+heparin 3500IU/ml) or lock solution with TauroLock-Hep500 (group 2: taurolidine citrate 4%+heparin 500 IU/ml) during the inter-dialysis period. The patients were allocated to these groups according to the hemodialysis center they were attending. RESULTS: A total of 145 CVCs were implanted in 127 patients and were followed for 15 months: 77 CVCs (65 patients) were placed in group 1 and 68 CVCs (62 patients) in group 2. There was no difference between the two groups with regard to CR-BSI (events per 1000 catheter-days: group 1=0.79, group 2=1.10; p=0.18) or exit site infection rates (events per 1000 catheter-days: group 1=2.45, group 2=1.83; p=0.37). The groups differed in ESI pathogens, with gram-positive oxacillin-resistant pathogens more frequent in group 1 (31.8% vs. 5.0%; p=0.003). The two groups were similar in mechanical complications. In the Cox regression analysis, the internal jugular vein site was a protective factor for all catheter removal complications (hazard ratio (HR) 0.41, 95% confidence interval (CI) 0.19-0.91) and mechanical complications (HR 0.16, 95% CI 0.065-0.41); only ESI was a risk factor for all catheter removal complications (HR 1.79, 95% CI 1.04-3.07) and mechanical complications (HR 5.64, 95% CI 1.65-19.3). CONCLUSIONS: The efficacy of both lock solutions was similar in preventing infections related to tunneled CVCs for HD. However, there were more oxacillin-resistant strains in patients who received antibiotic lock solution. Further studies are required to determine the optimal drug regimen and concentrations for lock solution and the associated adverse effects.

Unusual Intestinal Involvement by Paracoccidioidomycosis Diagnosed After Oral Manifestation.

Paracoccidioidomycosis is a common deep fungus infection in South America, particularly in Brazil. It is acquired through inhalation and primary involvement of lungs. Subsequently, dissemination may occur and oral mucosa is frequently affected and actually, in most of the cases the diagnosis is established because of the oral lesions. Thus, the role of the dentist is fundamental to correct diagnosis. However, the involvement of intestine is rarely reported. The current case describes a 36-year-old man who presented abdominal pain and intestinal constipation, being suspected and then confirmed as paracoccidioidomycosis after already be diagnosed with this disease by a dentist through oral manifestations.

Paracoccidioidomycosis Associated With a Heterozygous STAT4 Mutation and Impaired IFN-γ Immunity.

Background: Mutations in genes affecting interferon-γ (IFN-γ) immunity have contributed to understand the role of IFN-γ in protection against intracellular pathogens. However, inborn errors in STAT4, which controls interleukin-12 (IL-12) responses, have not yet been reported. Our objective was to determine the genetic defect in a family with a history of paracoccidioidomycosis. Methods: Genetic analysis was performed by whole-exome sequencing and Sanger sequencing. STAT4 phosphorylation (pSTAT4) and translocation to the nucleus, IFN-γ release by patient lymphocytes, and microbicidal activity of patient monocytes/macrophages were assessed. The effect on STAT4 function was evaluated by site-directed mutagenesis using a lymphoblastoid B cell line (B-LCL) and U3A cells. Results: A heterozygous missense mutation, c.1952 A>T (p.E651V) in STAT4 was identified in the index patient and her father. Patient's and father's lymphocytes showed reduced pSTAT4, nuclear translocation, and impaired IFN-γ production. Mutant B-LCL and U3A cells also displayed reduced pSTAT4. Patient's and father's peripheral blood mononuclear cells and macrophages demonstrated impaired fungicidal activity compared with those from healthy controls that improved in the presence of recombinant human IFN-γ, but not rhIL-12. Conclusion: Our data suggest autosomal dominant STAT4 deficiency as a novel inborn error of IL-12-dependent IFN-γ immunity associated with susceptibility to paracoccidioidomycosis.

Effect of the Spiritist "passe" energy therapy in reducing anxiety in volunteers: A randomized controlled trial.

BACKGROUND: In Brazil, one of the most used energy therapies is the Spiritist "passe", which is practiced by Spiritist healers. Although experimental studies have demonstrated the effectiveness of different energy therapies in reducing anxiety and pain, little is known about the effect of the Spiritist "passe" on health outcomes. Therefore, the present study aimed at evaluating the effectiveness of "passe" energy therapy in reducing anxiety symptoms. METHODS/DESIGN: In this prospective, randomized controlled trial, participants were randomly allocated into two groups: Intervention (8 weekly Spiritist "passe" sessions, n=23), and Control (8 weekly sham Spiritist "passe" sessions, n=27). Anxiety was assessed using the Trait Anxiety Inventory (STAI-trait). RESULTS: Of 97 individuals screened, 50 were included in the final analysis. Decreasing STAI-trait anxiety scores were observed in both groups throughout the study (p<0.0001). By the end of the study, 17% and 63% of intervention and control participants, respectively, still met the criterion for anxiety (p=0.001). However, anxiety reduction was more accentuated in the Spiritist "passe" group (p=0.02). CONCLUSION: In this small pilot study, anxiety was more markedly reduced in participants receiving the Spiritist "passe" than in controls, warranting larger trials. TRIAL REGISTRATION: NCT02376959.

Immunoreactivity of synthetic peptides derived from proteins of Cryptococcus gattii.

AIM: To determine the immunoreactivity of synthetic Cryptococcus-derived peptides. MATERIALS & METHODS: A total of 63 B-cell epitopes from previously identified Cryptococcus gattii immunoreactive proteins were synthesized and evaluated as antigens in ELISAs. The peptides were first evaluated for their ability to react against sera from immunocompetent subjects carrying cryptococcal meningitis. Peptides that yielded high sensitivity and specificity in the first test were then retested with sera from individuals with other fungal pathologies for cross-reactivity determination. RESULTS: Six of 63 synthetic peptides were recognized by antibodies in immunoassays, with a specificity of 100%, sensitivity of 78% and low cross-reactivity. CONCLUSION: We successfully determined the immunoreactivity of selected synthetic peptides of C. gattii derived proteins.