Search

Loop-mediated isothermal amplification (LAMP) is a highly effective molecular diagnostic technique, particularly advantageous for point-of-care (POC) settings. In recent years, LAMP has expanded to include various adaptations such as DARQ-LAMP, QUASR, FLOS-LAMP, displacement probes and molecular beacons. These methods enable multiplex detection of multiple targets in a single reaction, enhancing cost-effectiveness and diagnostic efficiency. Consequently, LAMP has gained significant traction in diagnosing diverse viruses, notably during the COVID-19 pandemic. However, its application for detecting Herpesviridae remains relatively unexplored. This group of viruses is of particular interest due to their latency and potential reactivation, crucial for immunocompromised patients, including organ and hematopoietic stem cell transplant recipients. This review highlights recent advancements in LAMP for virus diagnosis and explores current research trends and future prospects, emphasizing the detection challenges posed by Herpesviridae.

Staphylococcus capitis has been recognized as a relevant opportunistic pathogen, particularly its persistence in neonatal ICUs around the world. Therefore, the aim of this study was to describe the epidemiological profile of clinical isolates of S. capitis and to characterize the factors involved in the persistence and pathogenesis of these strains isolated from blood cultures collected in a hospital in the interior of the state of São Paulo, Brazil. A total of 141 S. capitis strains were submitted to detection of the mecA gene and S CCmec typing by multiplex PCR. Genes involved in biofilm production and genes encoding enterotoxins and hemolysins were detected by conventional PCR. Biofilm formation was evaluated by the polystyrene plate adherence test and phenotypic resistance was investigated by the disk diffusion method. Finally, pulsed-field gel electrophoresis (PFGE) was used to analyze the clonal relationship between isolates. The mecA gene was detected in 99 (70.2%) isolates, with this percentage reaching 100% in the neonatal ICU. SCC mec type III was the most prevalent type, detected in 31 (31.3%) isolates and co-occurrence of SCC mec was also observed. In vitro biofilm formation was detected in 46 (32.6%) isolates but was not correlated with the presence of the ica operon genes. Furthermore, biofilm production in ICU isolates was favored by hyperosmotic conditions, which are common in ICUs because of the frequent parenteral nutrition. Analysis of the clonal relationship between the isolates investigated in the present study confirms a homogeneous profile of S. capitis and the persistence of clones that are prevalent in the neonatal ICU and disseminated across the hospital. This study highlights the adaptation of isolates to specific hospital environments and their high clonality.

The expression of prostate-specific membrane antigen (PSMA) is upregulated in prostate cancer (PCa) cells and PSMA-ligands have been radiolabeled and used as radiopharmaceuticals for targeted radionuclide therapy (TRT), single photon emission computed tomography (SPECT) or positron emission tomography (PET) molecular imaging, and radioguided surgery in PCa patients. Herein, we aimed at radiolabeling the PSMA-I&S cold kit with 99m Tc, resulting in a radiopharmaceutical with high radiochemical yield (RCY) and stability for SPECT imaging and radioguided surgery in PCa malignancies. Various pre-clinical assays were conducted to evaluate the [ 99m Tc]Tc-PSMA-I&S obtained by the cold kit. These assays included assessments of RCY, radiochemical stability in saline, lipophilicity, serum protein binding (SPB), affinity for LNCaP-PCa cells (binding and internalization studies), and ex vivo biodistribution profile in naive and LNCaP-PCa-bearing mice. The radiopharmaceutical was obtained with good RCY (92.05% ± 2.20%) and remained stable for 6 h. The lipophilicity was determined to be -2.41 ± 0.06, while the SPB was ∼97%. The binding percentages to LNCaP cells were 9.41% ± 0.57% (1 h) and 10.45% ± 0.45% (4 h), with 63.12 ± 0.93 (1 h) and 65.72% ± 1.28% (4 h) of the bound material being internalized. Blocking assays, employing an excess of unlabeled PSMA-I&S, resulted in a reduction in the binding percentage by 2.6 times. The ex vivo biodistribution profile confirmed high accumulation of [ 99m Tc]Tc-PSMA-I&S in the tumor and the tumor-to-contralateral muscle ratio was ∼6.5. In conclusion, [ 99m Tc]Tc-PSMA-I&S was successfully obtained by radiolabeling the cold kit using freshly eluted [ 99m Tc]NaTcO 4 , exhibiting good RCY and radiochemical stability. The preclinical assays demonstrated that the radiopharmaceutical shows favorable characteristics for SPECT imaging and radioguided surgery in PCa patients., Competing Interests: SG was employed by Tecnonuclear-Eckert Ziegler The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Fuscaldi, Sobral, Durante, Mendonça, Miranda, Salgueiro, de Castiglia, Yamaga, da Cunha, Malavolta, de Barboza and Mejia.)

This study aimed to explore the molecular epidemiology of Staphylococcus aureus isolated from patients on mechanical ventilation and the participation of virulence factors in the development of ventilator-associated pneumonia (VAP). A prospective cohort study was conducted on patients under mechanical ventilation, with periodic visits for the collection of tracheal aspirates and clinical data. The S. aureus isolates were analyzed regarding resistance profile, virulence, expression of protein A and alpha-toxin using Western blot, clonal profile using PFGE, sequence type using MLST, and characterization and quantification of phenol-soluble modulins. Among the 270 patients in the study, 51 S. aureus strains were isolated from 47 patients. The incidence density of S. aureus and MRSA VAP was 2.35/1000 and 1.96/1000 ventilator days, respectively; of these, 45% (n = 5) were resistant to oxacillin, with 100% (n = 5) harboring SCC mec types II and IV. The most frequent among the tested virulence factors were ica A, hla , and hld . The clonal profile showed a predominance of sequence types originating from the community. Risk factors for VAP were the presence of solid tumors and the sea gene. In conclusion, patient-related risk factors, together with microbiological factors, are involved in the development of S. aureus VAP, which is caused by the patient's own strains.

The emergence of Community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) infections among indigenous populations has been reported. Usually, indigenous communities live in extreme poverty and are at risk of acquiring infections. In Brazil, healthcare inequality is observed in this population. To date, there are no reports of CA-MRSA infections, and no active search for asymptomatic S. aureus carriage has been conducted among Brazilian Indians. The aim of this study was to investigate the prevalence of colonization with S. aureus and CA-MRSA among Brazilian Indians. We screened 400 Indians (from near urban areas and remote hamlets) for S. aureus and CA-MRSA colonization. The isolates were submitted to clonal profiling by pulsed-field gel electrophoresis (PFGE), and selected isolates were submitted to multilocus sequence typing (MLST). Among 931 specimens (nasal and oral) from different indigenous individuals in remote hamlets, S. aureus was cultured in 190 (47.6%). Furthermore, CA-MRSA was found in three isolates (0.7%), all SCC mec type IV. PFGE analysis identified 21 clusters among the S. aureus isolates, and MLST analysis showed a predominance of sequence type 5 among these isolates. Our study revealed a higher prevalence of S. aureus carriage among Shanenawa ethnicity individuals (41.1%). Therefore, ethnicity appears to be associated with the prevalence of S. aureus in these populations.

At present, multidrug-resistant microorganisms are already responsible for community-acquired infections. Methicillin-resistant Staphylococcus aureus (MRSA) poses a serious public health risk worldwide because of the rapid spread and diversification of pandemic clones that are characterized by increasing virulence and antimicrobial resistance. The aim of this study was to identify the prevalence and factors associated with nasal, oral and rectal carriage of S. aureus and MRSA in bedridden patients and residents of long-term care facilities for the elderly (LTCFs) in Botucatu, SP, Brazil. Nasal, oral and rectal swab isolates obtained from 226 LTCF residents or home-bedridden patients between 2017 and 2018 were submitted to susceptibility testing, detection of the mecA gene, SCCmec characterization, and molecular typing by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). Logistic regression analysis was used to identify risk factors associated with the presence of S. aureus and MRSA. The prevalence of S. aureus and MRSA was 33.6% (n = 76) and 8% (n = 18), respectively. At the nine LTCFs studied, the prevalence of S. aureus ranged from 16.6% to 85.7% and that of MRSA from 13.3% to 25%. Living in an LTCF, male gender, a history of surgeries, and a high Charlson Comorbidity Index score were risk factors associated with S. aureus carriage, while MRSA carriage was positively associated with male gender. This study showed a high prevalence of S. aureus among elderly residents of small (<15 residents) and medium-sized (15−49 residents) LTCFs and a higher prevalence of MRSA in the oropharynx.

(1) Background: Peritonitis due to nonfermenting Gram-negative bacilli (NF-GNB) is a dramatic complication of peritoneal dialysis (PD) with bad outcomes. Previous studies of PD-related peritonitis due to Pseudomonas species have shown a low-resolution rate, without a high resistance rate to antipseudomonal antibiotics. This suggests that bacterial virulence factors can act and influence peritonitis evolution. This study aimed to describe the microbiological characteristics of NF-GNB causing PD-related peritonitis and analyze their influence on the outcome. (2) Methods: We analyze the 48 isolates from NF-GNB peritonitis, which were stored in our culture collection regarding bacterial resistance, biofilm, and other virulence factors' production, and clonal profile. Additionally, we collected data on treatment and outcomes from patients' clinical registers. (3) Results: The etiologies were species of Pseudomonas (50%), Acinetobacter (36%), and other NF-GNB (14%). There was a high (75%) proportion of biofilm producer lineages. The in vitro susceptibility rate of Pseudomonas spp. to amikacin, ciprofloxacin, and ceftazidime was significantly greater than that of Acinetobacter spp. and other species; however, there was a similar low-resolution rate (<45%) among the episodes attributable to them. Pseudomonas species have a polyclonal profile, while we found a clone of five multiresistant Acinetobacter baumannii over an 8-year interval (2000-2008), which suggest an origin from the healthcare environment. (4) Conclusions: We are not able to identify any predictor of outcome, but it is possible that biofilm and others virulence factors can act in concert and contribute to the bad outcome.

Coagulase-negative staphylococci (CoNS) may be considered contaminants when isolated from clinical specimens but may also be a cause of true infection. This study aimed to compare the clonality and SCC mec type of a collection of CoNS isolated from blood cultures of inpatients, nasal swabs of healthy individuals, and patients with chronic wounds, all from the same community, using SCC mec typing, pulsed-field gel electrophoresis (PFGE), and MLST. Staphylococcus epidermidis , exhibited high clonal diversity, but hospital and community clusters were observed. Nosocomial S. epidermidis clones belonged to sequence types ST2, ST6, and ST23. Some Staphylococcus haemolyticus clones were found to circulate in the hospital and community, while Staphylococcus saprophyticus exhibited very high clonal diversity. Staphylococcus lugdunensis , Staphylococcus warneri , and Staphylococcus capitis revealed several isolates belonging to the same clone in the hospital and community. The detection of different SCC mec types within the same cluster indicated high diversity. S. epidermidis was associated with SCC mec I and III, S. haemolyticus with I and II, S. capitis with type V, Staphylococcus hominis with mec complex type A and ccr1 , and S. warneri and S. saprophyticus with SCC mec I. The generation of elements and new combinations of cassette genes were highly associated with CoNS isolates, suggesting that SCC mec may not be a good marker of clonality in these bacteria.

Prostate-specific membrane antigen (PSMA) is a glycoprotein present in the prostate, that is overexpressed in prostate cancer (PCa). Recently, PSMA-directed radiopharmaceuticals have been developed, allowing the pinpointing of tumors with the Positron Emission Tomography (PET) or Single Photon Emission Computed Tomography (SPECT) imaging techniques. The aim of the present work was to standardize and validate an automatic synthesis module-based radiolabeling protocol for [ 68 Ga]Ga-PSMA-11, as well as to produce a radiopharmaceutical for PET imaging of PCa malignancies. [ 68 Ga]Ga-PSMA-11 was evaluated to determine the radiochemical purity (RCP), stability in saline solution and serum, lipophilicity, affinity to serum proteins, binding and internalization to lymph node carcinoma of the prostate (LNCaP) cells, and ex vivo biodistribution in mice. The radiopharmaceutical was produced with an RCP of 99.06 ± 0.10%, which was assessed with reversed-phase high-performance liquid chromatography (RP-HPLC). The product was stable in saline solution for up to 4 h (RCP > 98%) and in serum for up to 1 h (RCP > 95%). The lipophilicity was determined as -3.80 ± 0.15, while the serum protein binding (SPB) was <17%. The percentages of binding to LNCaP cells were 4.07 ± 0.51% (30 min) and 4.56 ± 0.46% (60 min), while 19.22 ± 2.73% (30 min) and 16.85 ± 1.34% (60 min) of bound material was internalized. High accumulation of [ 68 Ga]Ga-PSMA-11 was observed in the kidneys, spleen, and tumor, with a tumor-to-contralateral-muscle ratio of >8.5 and a tumor-to-blood ratio of >3.5. In conclusion, an automatic synthesis module-based radiolabeling protocol for [ 68 Ga]Ga-PSMA-11 was standardized and the product was evaluated, thus verifying its characteristics for PET imaging of PCa tumors in a clinical environment.

Background: People with diabetes mellitus, especially insulin-dependent diabetic patients, are a risk group for staphylococcal infections. Asymptomatic infection with Staphylococcus aureus is common and favors dissemination of the microorganism, rendering these individuals a source of infection. This study aimed to characterize the resistance profile, clonal profile and sequence type, as well as to analyze the prevalence and risk factors for nasal and oropharyngeal carriage of methicillin-susceptible (MSSA) and methicillin-resistant S. aureus (MRSA) isolated from insulin-dependent diabetic individuals in the city of Botucatu, SP, Brazil., Methods: Staphylococcus aureus was collected from the nasopharynx and oropharynx of 312 community-dwelling insulin-dependent diabetic individuals over a period of 3 years (October 2015 to December 2018). The isolates were characterized by susceptibility profiling, detection of the mecA gene, SCCmec typing, and molecular typing by PFGE and MLST. The risk factors associated with S. aureus and MRSA carriage were determined by logistic regression analysis., Results: The overall prevalence of colonization with S. aureus and MRSA was 30.4% and 4.8%, respectively. Fifteen of the 112 S. aureus isolates carried the mecA gene; SCCmec type IV was identified in 10 isolates, SCCmec type I in three, and SCCmec type II in two. Among the 15 resistant isolates (MRSA), four were susceptible to oxacillin/cefoxitin by the disc diffusion method and one MSSA isolate was resistant to sulfamethoxazole/trimethoprim. The analysis of risk factors revealed a protective effect of age and lung disease, while lower-extremity ulcers were a risk factor for S. aureus. For MRSA, only male gender was significantly associated as a risk factor in multivariate analysis. Clonal profile analysis demonstrated the formation of clusters among MRSA isolates from different patients, with the identification of ST5-IV, ST5-I, and ST8-IV. Isolates carrying ST398 were identified among MSSA and MRSA (ST398-IV)., Conclusion: Our findings reinforce the importance of epidemiological studies of S. aureus carriage, especially in populations at high risk of infections such as diabetics. The data suggest widespread dissemination of MRSA in the population of insulin-dependent diabetic patients studied, as well as the emergence of important lineages among these individuals.

Bovine mastitis is mainly caused by bacteria of the genus Staphylococcus spp., which possess different virulence factors, including the capacity for biofilm formation that provides enhanced protection against the action of immune system components and serves as a barrier against the penetration of antimicrobial agents. This study aimed to characterize 181 Staphylococcus spp. Strains-including Staphylococcus aureus and coagulase-negative staphylococci (CoNS) isolated from bovine subclinical mastitis in six Brazilian states-by molecular methods. RT-qPCR was used to verify the expression of genes of the ica operon-mainly responsible for biofilm formation-as well as bap and bhp . Chromosome similarity among the isolates was investigated by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). The ica A gene was detected in 79 (43.6%) isolates, ica B in 24 (13.2%), ica C in 57 (31.4%), and ica D in 127 (70.1%). The bap gene was identified in 66 (36.4%) isolates, while the bhp gene was found in nine (4.9%). RT-qPCR confirmed the expression of the ica A gene in 60 (75.9%) isolates, of ica B in six (25%), of ica C in 26 (45.6%), and of ica D in 80 (63%). Clonal typing of the isolates by PFGE permitted the identification of eight Staphylococcus aureus clusters that simultaneously included ≥3 strains, with a similarity of ≥80%. Regarding the other species studied, three clusters were observed for Staphylococcus chromogenes and four clusters for Staphylococcus epidermidis . Only one cluster each was identified for Staphylococcus saprophyticus and Staphylococcus simulans , while the other species did not form any cluster. With respect to MLST, ST126 and ST1 were the prevalent sequence types in S. aureus , while in S. epidermidis all sequence types were different. These results reveal strains with the same evolutionary origin as other isolates, which might cause infections in humans and animals, suggesting their ability to spread between these species.

The ability of Staphylococcus epidermidis to produce virulence factors, such as biofilm, added to its increased resistance to antimicrobials can cause infections that are difficult to treat. Many staphylococcal virulence factors are under the control of the accessory gene regulator ( agr ). The objective of this study was to establish the agr locus and susceptibility of biofilm-producing S. epidermidis specimens to antimicrobial agents, through PCR reactions, reverse transcription polymerase chain reaction (RT-PCR), and the determination of minimum inhibitory concentration (MIC), and to analyze the clonal profile of 300 strains isolated from blood culture specimens from inpatients at a University Hospital in Brazil, over a 20-year period by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) techniques. The ica operon expression was shown in 83.6% strains, bhp gene in 11.5%, and aap gene in 32.8%. Oxacillin resistance was detected in 90.1%, while 4.9% showed tigecycline resistance, and intermediate resistance to quinupristin/dalfopristin was identified in 0.4%. Clonal profile determination showed 11 clusters, with the ST2 type determined as the major cluster. The S. epidermidis biofilm producer demonstrated a predominance of agr I locus, oxacillin resistance, and SCC mec III as well as the potential dissemination of pathogenic clones in hospital settings over long periods.

Background: The positron emission tomography (PET) ligand 68 Ga-Glu-urea-Lys(Ahx)-HBED-CC ( 68 Ga-PSMA-11) targets the prostate-specific membrane antigen (PSMA), upregulated in prostate cancer cells. Although 68 Ga-PSMA-11 PET is widely used in research and clinical practice, full kinetic modeling has not yet been reported nor have simplified methods for quantification been validated. The aims of our study were to quantify 68 Ga-PSMA-11 uptake in primary prostate cancer patients using compartmental modeling with arterial blood sampling and to validate the use of standardized uptake values (SUV) and image-derived blood for quantification., Results: Fifteen patients with histologically proven primary prostate cancer underwent a 60-min dynamic 68 Ga-PSMA-11 PET scan of the pelvis with axial T1 Dixon, T2, and diffusion-weighted magnetic resonance (MR) images acquired simultaneously. Time-activity curves were derived from volumes of interest in lesions, normal prostate, and muscle, and mean SUV calculated. In total, 18 positive lesions were identified on both PET and MR. Arterial blood activity was measured by automatic arterial blood sampling and manual blood samples were collected for plasma-to-blood ratio correction and for metabolite analysis. The analysis showed that 68 Ga-PSMA-11 was stable in vivo. Based on the Akaike information criterion, 68 Ga-PSMA-11 kinetics were best described by an irreversible two-tissue compartment model. The rate constants K 1 and k 3 and the net influx rate constants K i were all significantly higher in lesions compared to normal tissue (p < 0.05). K i derived using image-derived blood from an MR-guided method showed excellent agreement with K i derived using arterial blood sampling (intraclass correlation coefficient = 0.99). SUV correlated significantly with K i with the strongest correlation of scan time-window 30-45 min (rho 0.95, p < 0.001). Both K i and SUV correlated significantly with serum prostate specific antigen (PSA) level and PSA density., Conclusions: 68 Ga-PSMA-11 kinetics can be described by an irreversible two-tissue compartment model. An MR-guided method for image-derived blood provides a non-invasive alternative to blood sampling for kinetic modeling studies. SUV showed strong correlation with K i and can be used in routine clinical settings to quantify 68 Ga-PSMA-11 uptake.

Objective: Positron emission tomography in association with magnetic resonance imaging (PET/MR) and 68 Ga-PSMA-11 has shown superior detection in recurrent prostate cancer patients as compared to PET/computed tomography (PET/CT). There are, however, several technological differences between PET/CT and PET/MR systems which affect the PET image quality. The objective of this study was to assess the reproducibility of PET/CT and PET/MR SUV's in recurrent prostate cancer patients. We randomized the patients regarding the order of the PET/CT and PET/MR scans to reduce the influence of tracer uptake as a function of time., Methods: Thirty patients, all with biochemical recurrence after radical prostatectomy, underwent whole-body PET/CT and PET/MR scans after intravenous injection of a single dose of 68 Ga-PSMA-11. Fifteen patients underwent PET/CT first and 15 patients underwent PET/MR first. Volumes of interest on tumor lesions were outlined and maximum standardized uptake value (SUVmax) corrected for lean body mass was calculated. Correlation and agreement between scans were assessed by generalized linear mixed-effects models and Bland-Altman analysis. The association between SUV, patient characteristics and imaging parameters was assessed., Results: Eighteen of the 30 evaluated patients had at least one positive lesion, giving an overall detection rate of 60%. In total, there were 34 visible lesions: 5 local recurrences, 22 lymph node metastases and 7 bone metastases. One group acquired PET/CT and PET/MR at median time points of 63.0 and 159.0 min, while the other group acquired PET/MR and PET/CT at median time points of 92.0 and 149.0 min. SUVmax between scans was linearly correlated, described by the equation Y(PET/CT SUVmax) = 0.75 + 1.00 × (PET/MR SUVmax), on average 20% higher on PET/CT than on PET/MR. SUV associated significantly only with type of lesion, scan time post-injection and acquisition time per bed position., Conclusions: SUVmax from PET/CT and PET/MR are linearly correlated, on average 20% higher on PET/CT than on PET/MR and should, therefore, not be used interchangeably in patient follow-up.

The use of antimicrobial agents has led to the emergence of resistant bacterial strains over a relatively short period. Furthermore, Staphylococcus spp. can produce β-lactamase, which explains the survival of these strains in a focus of infection despite the use of a β-lactam antibiotic. The aim of this study was to evaluate the resistance of Staphylococcus spp. isolated from bovine subclinical mastitis to oxacillin and vancomycin (by minimum inhibitory concentration) and to detect vancomycin heteroresistance by a screening method. We also evaluated β-lactamase production and resistance due to hyperproduction of this enzyme and investigated the mecA and mecC genes and performed staphylococcal cassette chromosome mec typing. For this purpose, 181 Staphylococcus spp. isolated from mastitis subclinical bovine were analyzed. Using the phenotypic method, 33 (18.2%) of Staphylococcus spp. were resistant to oxacillin. In contrast, all isolates were susceptible to vancomycin, and heteroresistance was detected by the screening method in 13 isolates. Production of β-lactamase was observed in 174 (96%) of the Staphylococcus spp. isolates. The mecA gene was detected in 8 isolates, all of them belonging to the species Staphylococcus epidermidis, and staphylococcal cassette chromosome mec typing revealed the presence of type I and type IV isolates., (Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Staphylococcus aureus and Staphylococcus saprophyticus are the most common and most important staphylococcal species associated with urinary tract infections. The objective of the present study was to compare and to evaluate the accuracy of four phenotypic methods for the detection of beta-lactamase production in Staphylococcus spp. Seventy-three strains produced a halo with a diameter ≤28mm (penicillin resistant) and all of them were positive for the blaZ gene. Among the 28 susceptible strain (halo ≥29mm), 23 carried the blaZ gene and five did not. The zone edge test was the most sensitive (90.3%), followed by MIC determination (85.5%), but the specificity of the former was low (40.0%). The nitrocefin test was the least sensitive (28.9%). However, the nitrocefin test together with the disk diffusion method showed the highest specificity (100%). The present results demonstrated that the zone edge test was the most sensitive phenotypic test for detection of beta-lactamase, although it is still not an ideal test to detect this type of resistance since its specificity was low. However, the inhibition halo diameter of the penicillin disk can be used together with the zone edge test since the same disk is employed in the two tests. Combined analysis of the two tests shows a sensitivity of 90.3% and specificity of 100%, proving better sensitivity, especially for S. saprophyticus. This is a low-cost test of easy application and interpretation that can be used in small and medium-sized laboratories where susceptibility testing is usually performed by the disk diffusion method., (Copyright © 2016 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.)

Objetive: to characterize the care services performed through risk rating by the Manchester Triage System, identifying demographics (age, gender), main flowcharts, discriminators and outcomes in pediatric emergency., Method: cross-sectional quantitative study. Data on risk classification were obtained through a search of computerized registration data from medical records of patients treated in the pediatric emergency within one year. Descriptive statistics with absolute and relative frequencies was used for the analysis., Results: 10,921 visits were conducted in the pediatric emergency, mostly male (54.4%), aged between 29 days and two years (44.5%). There was a prevalence of the urgent risk category (43.6%). The main flowchart used in the care was worried parents (22.4%) and the most prevalent discriminator was recent event (15.3%). The hospitalization outcome occurred in 10.4% of care performed in the pediatric emergency, however 61.8% of care needed to stay under observation and / or being under the health team care in the pediatric emergency., Conclusion: worried parents was the main flowchart used and recent events the most prevalent discriminator, comprising the hospitalization outcomes and permanency in observation in the pediatric emergency before discharge from the hospital.

Invasive aspergillosis is the primary opportunistic invasive fungal infection described in neutropenic hematologic patients, caused by the angioinvasive pathogen Aspergillus fumigatus. The molecular mechanisms associated with A. fumigatus infection in the vascular endothelium are poorly understood. In this context, we used a high-throughput proteomic approach to unveil the proteins modulated in HUVECs after interaction with a wild type strain and the UGM1 mutant (Δugm1) of A. fumigatus. The proteomic analysis was also performed in HUVECs challenged with a galactosaminogalactan (GAG) purified from A. fumigatus cell wall. The dataset presented here correspond to all proteins identified that fit a 2-fold change criteria (log 2 ratio ≥ 1 or ≤ -1), disregarding the statistical validation cut off, in order to supplement the research article entitled "Modifications to the composition of the hyphal outer layer of Aspergillus fumigatus modulates the HUVEC proteins associated with inflammatory and stress responses" (G.W.P. Neves, N.A. Curty, P.H. Kubitschek-Barreira, T. Fontaine, G.H.M.F. Souza, M. Lyra Cunha, G.H. Goldman, A. Beauvais, J.P. Latgé, L.M. Lopes-Bezerra, 2016) [1]. The mass spectrometry proteomic data have been deposited in the ProteomeXchange Consortium via the PRIDE partner repository with the dataset identifier PRIDE: PXD002823.

Objective: To understand the experiences of nurses when caring for dying newborns and their families in the NICU; and redeem their perceptions about acting before the death and grieving process., Method: A descriptive exploratory study with a qualitative approach, developed with nine nurses at the ICU of a hospital in São Paulo (SP), Brazil. Data was collected through semi-structured interviews and analyzed using the Collective Subject Discourse (CSD)., Results: Caring for newborns who are dying and their families is very difficult for nurses, due to the intense involvement. They seek strategies to deal with the situation and, before the newborn's death, despite the suffering, express the feeling of accomplishment., Conclusions: Facing death and grief triggers mechanisms that emerge life references, coming across painful issues. Learning to deal with these questions is a daily challenge for nurses of the NICU., Objetivo: Compreender as experiências vivenciadas por enfermeiros ao cuidar de neonatos que estão morrendo e seus familiares na UTIN; e resgatar as suas percepções sobre a atuação diante do processo de morte e luto., Método: Estudo descritivo exploratório, de abordagem qualitativa, desenvolvido com nove enfermeiras da UTIN de um hospital de São Paulo (SP), Brasil. Os dados foram coletados por meio de entrevista semi-estruturada e analisados pela técnica do Discurso de Sujeito Coletivo (DSC)., Resultados: Cuidar de neonatos que estão morrendo e suas famílias é muito difícil para as enfermeiras, devido ao intenso envolvimento. Buscam estratégias para lidar com a situação e, diante do óbito do neonato, apesar do sofrimento, manifestam o sentimento de dever cumprido., Conclusión: Enfrentar a morte e o luto aciona mecanismos que afloram referências de vida, deparando-se com questões dolorosas. Aprender a lidar com essas questões é um desafio diário para os enfermeiros de UTIN.

Objective: The research objective was to characterize preterm infants with birth weight under 1500 g, and to identify the incidence of late-onset neonatal sepsis among this population., Methods: A prospective cohort study with a sample of 30 preterm newborns that weighed under 1500 g and were hospitalized in the NICU of the university hospital. Data were collected from January to December 2013 using a structured instrument., Results: Of the 30 neonates included in the study, 14 developed late-onset neonatal sepsis with a prevalence of coagulase-negative staphylococci., Conclusions: The incidence of late-onset neonatal sepsis indicates a vulnerability in preterm infants due to immunological immaturity. These results reveal that knowledge of the profile of newborn infants admitted to the NICU and the risk factors to which they are exposed are central to the planning of nursing care for these patients. Future studies should address strategies for preventing nosocomial infection.

Objective: To evaluate the perception of healthcare professionals about the safety culture in the operating room of a public hospital, large-sized, according to the domains of the Safety Attitudes Questionnaire (SAQ)., Method: Descriptive, cross-sectional and quantitative research, with the application of the SAQ to 226 professionals. Descriptive data analysis, instrument consistency and exploratory factor analysis., Results: Participants were distributed homogeneously between females (49.6%) and males (50.4%); mean age of 39.6 (SD±9.9) years and length of professional experience of 9.9 (SD ± 9.2) years. And Cronbach's α of 0.84. It was identified six domains proposed in the questionnaire: stress perception (74.5) and job satisfaction (70.7) showed satisfactory results; teamwork environment (59.1) and climate of security (48.9) presented scores below the minimum recommended (75); unit's management perceptions (44.5), hospital management perceptions (34.9) and working conditions (41.9) presented the lowest averages., Conclusions: The results showed that, from the perspective of the professionals, there is weakness in the values, attitudes, skills and behaviors that determine the safety culture in a healthcare organization.

Context: The Neonatal Infant Pain Scale (NIPS), initially developed in Canada, has been previously used but not adequately adapted and validated for use in Brazil., Objectives: The goal of the present study was to perform a cross-cultural adaptation and clinical validation of the NIPS for use in the Brazilian population., Methods: The instrument was adapted based on the method outlined by Beaton et al., including the production and combination of translated versions, back-translation, committee review, and pilot testing. The psychometric properties of the adapted instrument, including its validity, reliability, and internal consistency, were tested in a clinical validation study. The sample comprised 60 at-term newborns who were evaluated by six nurses as they experienced vaccination. Psychometric properties were evaluated using Student's t-tests, prevalence-adjusted and bias-adjusted kappa scores, the Bland-Altman method, and Cronbach's alpha coefficients., Results: The Brazilian version of the NIPS (Escala de Dor no Recém-Nascido [NIPS-Brazil]) demonstrated excellent interobserver and intraobserver reliability. Total NIPS-Brazil scores yielded prevalence-adjusted and bias-adjusted kappa scores of 0.93, whereas the Bland-Altman method revealed interobserver and intraobserver reliability values of 95% and 90%, respectively. The NIPS-Brazil had adequate internal consistency, as evidenced by a Cronbach's alpha of 0.762., Conclusion: The NIPS was successfully adapted for use in Brazil and is now available for use in the assessment of acute pain in at-term newborns in Brazil., (Copyright © 2015 American Academy of Hospice and Palliative Medicine. Published by Elsevier Inc. All rights reserved.)

Objective: To identify and compare behaviors of children during vaccination, who were prepared or not for the procedure using an instructional therapeutic play., Methods: A quasi experimental study, with quantitative approach of 60 children aged 3 to 6 years. The child's reactions were recorded in a checklist. Data were analyzed using descriptive and inferential statistics and Fisher's test., Results: The main reactions in the experimental group were stay still (25;83%) and spontaneously collaborate (24;80%). In the control group, the main reactions were cries and cling to parents (15; 50%), flushing (11;36.67%) and moving the body/agitated (10;33.3%)., Conclusion: The reactions of cooperation were more frequent in the experimental group, while low acceptance was observed only in the control group. Therapeutic play has proved an important tool in preparing for the vaccine.

We investigated a small Dutch family with a bleeding diathesis, prolonged prothrombin, and activated partial thromboplastin times, in whom no classifying diagnosis was made. The 2 affected relatives had severely decreased in vitro thrombin generation, and levels of tissue factor pathway inhibitor (TFPI) were strongly increased. To identify the genetic cause of the bleeding diathesis, we performed whole exome sequencing analysis of all living relatives. We found a novel gain-of-function mutation in the F5 gene (c.C2588G), which leads to an aberrant splicing of F5 and ultimately to a short factor V protein (missing 623 amino acids from the B domain), which we called factor V Amsterdam. Factor V Amsterdam binds to TFPI, prolonging its half-life and concentration. This is the second report of an association between a shorter form of factor V and increased TFPI levels, resulting in severely reduced thrombin generation and a bleeding tendency., (© 2015 by The American Society of Hematology.)

Objective: To describe the main non-pharmacological interventions for pain relief in newborns available in Neonatal Intensive Care Unit., Method: An exploratory search of the MedLine, Lilacs and Scielo online databases was conducted to retrieve references of studies published from 2004 to 2013., Results: Several non-pharmacological interventions were shown to be effective, to represent low risk for neonates and to have a low operational cost. The ones most often discussed in the literature were: oral administration of glucose/sucrose, non-nutritive sucking, breastfeeding, skin-to-skin contact, facilitated tucking and swaddling., Conclusion: Healthcare teams should be familiar with these methods and use them more effectively in Neonatal Intensive Care Unit daily routines, so as to ensure that newborns receive qualified and more human care.

Objective: To describe the process of cross-cultural adaptation and clinical validation of the Neonatal Skin Condition Score., Methods: This methodological cross-cultural adaptation study included five steps: initial translation, synthesis of the initial translation, back translation, review by an Committee of Specialists and testing of the pre-final version, and an observational cross-sectional study with analysis of the psychometric properties using the Adjusted Kappa, Intraclass Correlation Coefficient, and Bland-Altman Method statistical tests. A total of 38 professionals were randomly recruited to review the clarity of the adapted instrument, and 47 newborns hospitalized in the Neonatology Unit of the Clinical Hospital of Porto Alegre were selected by convenience for the clinical validation of the instrument., Results: The adapted scale showed approximately 85% clarity. The statistical tests showed moderate to strong intra and interobserver item to item reliability and from strong to very strong in the total score, with a variation of less than 2 points among the scores assigned by the nurses to the patients., Conclusions: The scale was adapted and validated to Brazilian Portuguese. The psychometric properties of the Brazilian version of the Neonatal Skin Condition Score instrument were similar to the validation results of the original scale.

The authors report the case of a 70-year-old male patient with chronic lymphoid leukemia who presented subsequently a papillary carcinoma of the thyroid with metastases to regional lymph nodes. The patient was treated with surgical thyroidectomy with regional and cervical lymph node excision and radioiodine therapy (I-131). The protocolar control scintigraphy 4 days after the radioactive dose showed I-131 uptake in both axillae and even in the inguinal regions. PET/CT showed faint FDG-F-18 uptake in one lymph node of the left axilla. An ultrasound guided fine needle biopsy of this lymph node identified by I-131 SPECT/CT and FDG-F-18 PET/CT revealed lymphoma cells and was negative for thyroid tissue and thyroglobulin content. The sequential blood counts done routinely after radiation treatment showed a marked fall until return to normal values of leucocytes and lymphocytes (absolute and relative), which were still normal in the last control 19 months after the radioiodine administration. Chest computed tomography showed a decrease in size of axillary and para-aortic lymph nodes. By immunohistochemistry, cells of the lymphoid B lineage decreased from 52% before radioiodine therapy to 5% after the procedure. The authors speculate about a possible sodium iodide symporter expression by the cells of this lymphoma, similar to some other non-thyroid tumors, such as breast cancer cells.

Objective: To compare vital signs before and after the therapeutic touch observed in hospitalized newborns in neonatal intensive care unit., Methods: This was a quasi-experimental study performed at a neonatal intensive care unit of a municipal hospital, in the city of São Paulo (SP), Brazil. The sample included 40 newborns submitted to the therapeutic touch after a painful procedure. We evaluated the vital signs, such as heart and respiratory rates, temperature and pain intensity, before and after the therapeutic touch., Results: The majority of newborns were male (n=28; 70%), pre-term (n=19; 52%) and born from vaginal delivery (n=27; 67%). Respiratory distress was the main reason for hospital admission (n=16; 40%). There was a drop in all vital signs after therapeutic touch, particularly in pain score, which had a considerable reduction in the mean values, from 3.37 (SD=1.31) to 0 (SD=0.0). All differences found were statistically significant by the Wilcoxon test (p<0.05)., Conclusion: The results showed that therapeutic touch promotes relaxation of the baby, favoring reduction in vital signs and, consequently in the basal metabolism rate.

The objective of the current study was to isolate and identify phenotypes of Staphylococcus spp. strains derived from buffalo (Bubalus bubalis) milk. A total of 548 milk samples from 137 buffalo were cultured in Columbia agar enriched with 5% defibrinated sheep blood. Determination of the capacity of Staphylococcus aureus to produce enterotoxins A-D and toxic shock syndrome toxin-1 (TSST-1) was achieved by reverse passive latex agglutination (RPLA). Antimicrobial sensitivity of S. aureus strains was evaluated using the disk diffusion technique, and β-lactamase detection was achieved using the chromogenic test with paper discs impregnated with nitrocefin. From all the mammary quarters examined, 36 (10.8%) were positive for Staphylococcus spp., 83.3% were coagulase-negative staphylococcus (CNS), 11.1% were coagulase-positive staphylococcus (CPS), and 5.6% were of CPS+CNS positive. All isolates of S. aureus produced at least 1 toxin and 5 out of 6 isolates (83.0%) produced β-lactamase. One hundred percent of S. aureus isolates were sensitive to methicillin and amoxicillin + clavulanic acid, and resistant to ampicillin, penicillin, and oxacillin. Analysis of the results obtained in the current study highlight the epidemiologic importance of buffalo milk regarding the production of enterotoxins and TSST-1 and the potential risk to public health.

This study identifies the adverse events related to the use of central venous catheters (CVC) in newborns admitted to a neonatal care unit. This is a quantitative, descriptive and retrospective study. The population consisted of 167 newborns admitted in the neonatal unit of the Hospital de Clínicas at Porto Alegre, RS, Brazil which used CVCs inserted through percutaneous puncture (PICC) and surgical insertion, totaling 241 catheters. There was a higher prevalence of mechanical adverse events in the PICC line insertions, with a preponderance of catheter occlusions (19.44%) and ruptures (8.8%). The surgically inserted CVCs had a higher prevalence of catheter-related infectious adverse events with the most common being clinical sepsis (16%). This study suggests that the correct insertion technique should be used and a specialized team should monitor the CVCs to ensure safety and prevent adverse events.

After the prohibition of organochlorine-pesticide use in Brazil for controlling insect vector diseases, Mato Grosso State gathered the exceeding DDT and stored it irregularly in an open air area that belongs to the National Health Foundation, causing soil contamination. This study aimed to evaluate the contamination level and dissipation of p,p'-DDT and p,p'-DDE in this area. For that, surface soil samples were collected on 19 September 2000, 15 December 2000, 4 April 2001 and soil samples 30-40 cm; 60-70 cm and 90-100 cm deep were taken from five points in the studied area on 17 July 2001. The contaminants were determined by a small scale method which consists on extraction and clean-up steps combined into one step by transferring soil samples mixed with neutral alumina to a chromatographic column prepacked with neutral alumina and elution with hexane:dichloromethane (7:3 v:v). The eluate was concentrated and the analytes were quantified by gas chromatography with an electron-capture detector. p,p'-DDT at surface soil ranged from 3,800 to 7,300 mg kg(-1). 30-40 cm deep soil sample concentrations varied from 0.036 to 440 mg kg(-1) while 90-100 cm deep samples varied from 0.069 to 180 mg kg(-1). Volatilization is probably the main dissipation process. The p,p'-DDT is moving slowly downward in the soil profile, however, the levels of this contaminant are high enough to present risk to underground waters.

Objective: To assess the effect of bathing with water only or with mild pH neutral soap and water on skin flora of preterm newborns., Method: Randomized blinded clinical trial that enrolled 73 preterm newborns whose gestational age was between 28 and 35 weeks and birth weight between 800 and 1800 g. At the age of 3 days the infants were randomly assigned to a group that was given daily baths either with water only, or with soap and water for 7 or more days. Antibiotic treatment delayed onset of the trial in some preterm infants. On the final day of bathing axillary bacterial cultures were obtained before and 30 minutes after bathing., Results: At the time of axillary culturing, the mean age of the newborns was 19 days. Coagulase-negative Staphylococcus was the most prevalent microorganism in both groups. No significant difference was found in the count of microorganism colonies between both groups. The comparison of the number of bacterial colonies between the time before and after bath, carried out by repeated-measures ANOVA showed a significant difference over time in the two groups, without a significant difference between the two groups., Conclusions: Bathing preterm newborns with water only or with soap and water produces similar effects on skin colonization of a preterm neonate. Both are effective to decrease the number of colonies of Gram-positive and Gram-negative bacteria.

Although acute respiratory infections (ARIs) are a major cause of child morbidity and mortality in Southern Brazil, little information is available on their seasonality and viral etiology. This study was conducted on children under 5 years of age with ARI to assess viral etiology in the State of Rio Grande do Sul, from 1990 to 1992. A total of 862 nasopharyngeal secretion (NPS) samples were tested using indirect immunofluorescence. The results showed that 316 (36.6%) NPS samples were positive: 26.2% for RSV, 6% for adenovirus, 1.7% for influenza viruses, 1.5% for parainfluenza viruses, and 1.2% for mixed infection. The mean viral prevalence rates in out-patient services, emergency wards, and in-patient hospital wards were 26.7%, 53% and 42.3%, respectively. Respiratory syncytial virus (RSV) and adenovirus accounted for 91.4 % of the viral diagnoses. RSV was more frequent in children under one year of age at the three levels of health care and was prevalent in infants under six months. Adenovirus was the most prevalent pathogen in hospitalized children, in 1992. Influenza A virus showed an increased prevalence with age among out-patient children. This study shows the annual occurrence of viral respiratory infections in the coldest months, with a significant annual variation in the frequency of RSV infection.

In areas of drug-resistant malaria, control programs may restrict chemotherapy until malaria has been confirmed via microscopy to contain costs and toxicity. In Brazil, patients travel to centralized laboratory posts (FNS) at great cost for diagnosis and treatment. A program was established through the bars of a mining town offering free dipstick diagnosis and mefloquine treatment on a 24-hr basis; falciparum malaria dipstick tests are accurate and easy to use. Outcomes were compared with historical data and results of a neighboring non-intervention village. Guidelines for dipstick use and treatment were followed for 98% of visits. The number of FNS visits was reduced from 2,316 (expected) to 1,097 (observed) with 626 dipstick tests applied. Ninety-five percent of those who visited the FNS experienced onset of malaria symptoms in the town where the FNS was located. There was an unexpected doubling of the malaria hospital admission rate. We demonstrate that dipstick testing can be used in a sustainable, community-based program that should be applicable in a wide variety of settings.

OBJECTIVE: To evaluate the effect of cisapride and chest physical therapy on the gastroesophageal reflux of wheezing babies. METHODS: We prospectively assessed the presence of technetium ((99)Tc) in the upper, middle, and lower esophagus of 25 wheezing babies (13 with GERD and 12 without GERD) using scintigraphy. Both groups underwent clinical investigation, including laboratory, X-ray and scintigraphy tests, for the etiology of the wheezing baby syndrome (WBS) and GERD. Expiratory Flow Acceleration (EFA) was performed before and after treatment with cisapride. The total time of GER episodes was accounted for each portion of the esophagus during scintigraphy and during EFA. RESULTS: Cisapride significantly reduced the total reflux time in the upper esophagus (P<0.05), but showed no influence during EFA. After cisapride therapy, EFA increased the total reflux time in the upper and medium esophagus; however, no statistical significance was found. Infants with GERD presented a shorter total reflux time in the distal esophagus (P<0.05) during EFA. After cisapride treatment, no statistical significance was found. Infants without GERD also presented reduced total reflux time in the distal esophagus during EFA (P<0.05). Those with GERD had increased total reflux time in the distal esophagus (P<0.05) before and after cisapride treatment during EFA and scintigraphy. CONCLUSIONS: Cisapride was effective in reducing the total reflux time, mainly in the upper esophagus. EFA apparently increased the number of episodes of GER, without achieving statistical significance. Further studies are necessary to investigate the effects of chest physical therapy according to body positions.

Gender-related differences in gastric emptying are still controversial. The aims of this study were: to confirm the sex-related difference in gastric emptying of a solid meal and to investigate its association with different patterns of meal distribution between the proximal and distal gastric compartments. Eighteen healthy volunteers (nine males, mean age 35 +/- 9 years; nine females, mean age: 41 +/- 11 years) were studied in the morning, after ingestion of the solid test-meal (an omelette labeled with 185MBq of 99mTc-sulfur colloid). Simultaneous anterior and posterior images of the stomach were acquired immediately after ingestion of the meal and every 10 minutes for 120 minutes. Time versus activity curves were obtained for the whole, proximal and distal stomach. Gastric T1/2 was longer in women (96.1 +/- 17.2 min) than in men (79.9 +/- 17.8 min; P = 0.02). The analysis of the meal distribution inside the stomach showed no differences between males and females in proximal gastric emptying, but the meal retention in the distal compartment was significantly increased among women (P = 0.04). In conclusion, gastric emptying of a solid meal is slower in pre-menopausal women than in age-matched men, probably due to an increased retention of the meal in the distal compartment. This should be taken into consideration to avoid misleading diagnosis of gastroparesis for female patients.