Web-Based Virtual Microscopy for Parasitology: A Novel Tool for Education and Quality Assurance

Here, we describe a novel tool to observe parasites by virtual microscopy on the Internet. Microscopy-based identification of parasites is the basis for both diagnostics and epidemiological assessment of parasite burden globally. Yet, quality assessment of diagnostic parasitology laboratories is difficult, as delivering identical educational specimens has been impossible. In this study, a series of parasite specimens on ordinary glass slides were digitized using a recently developed microscope scanner technique. Up to 50,000 images captured at high magnification are digitally stitched together to form a representation of the entire glass slide. These “virtual slides” digitized at a thousand-fold magnification can hold more than 60 gigabytes of data. Handling such large amounts of data was made possible because of efficient compression techniques and a viewing system adopted from the geospatial imaging industry. Viewing the samples on the Internet very much resembles, for example, the use of Google Maps, and puts only modest requirements on the viewer's computer. In addition, we captured image stacks at different focal planes, and developed a web-based viewing system for three-dimensional navigation in the specimens. This novel technique is especially valuable for detailed visualization of large objects such as helminth eggs in stool specimens

Density Distribution of Pharyngeal Carriage of Meningococcus in Healthy Young Adults: New Approaches to Studying the Epidemiology of Colonization and Vaccine Indirect Effects.

BACKGROUND: Improved understanding of Neisseria meningitidis (Nm) carriage biology and better methods for detection and quantification would facilitate studies of potential impact of new vaccines on colonization and transmission in adolescents. METHODS: We performed plate cultures on 107 oropharyngeal swabs stored frozen in skim milk tryptone glucose glycerol (STGG) broth and previously positive for Nm. We compared quantitative polymerase chain reaction (qPCR) detection of Nm in 601 STGG-swabs with culture. Using qPCR (n = 87), a log-phase broth culture standard curve and semiquantitative plate cultures (n = 68), we measured density of carriage. We compared qPCR genogrouping of DNA extracts from STGG-swabs and from plate culture lawns (n = 110) with purified isolates (n = 80). RESULTS: Swab storage resulted in only 10% loss of culture sensitivity. Direct sodC qPCR Nm detection yielded more positives (87/601, 14.5%) than culture (80/601, 13.3%). Most samples (57/110) positive by culture were also positive by qPCR and vice versa, but discrepancies (single positives) were frequent among low-density samples. sodC qPCR was positive in 79/80 isolates but in only 65 by ctrA qPCR. Density both by culture and qPCR varied across 4 orders of magnitude with the majority being low (1000). Genogrouping qPCRs yielded more positive results when performed on DNA extracts from lawn cultures. CONCLUSIONS: We provide the first description of the distribution of Nm carriage density. This could be important for understanding transmission dynamics and population-level effectiveness of adolescent vaccine programs. Storage of swabs frozen in STGG for batched laboratory analysis facilitates carriage studies and direct sodC qPCR for Nm combined with qPCR genogrouping of lawn culture extracts provides accurate, detailed description of colonization

Henoch-Schönlein nephritis associated with streptococcal infection and persistent hypocomplementemia: a case report

<p>Abstract</p> <p>Introduction</p> <p>Henoch-Schönlein purpura is a systemic disease with frequent renal involvement, characterized by IgA mesangial deposits. Streptococcal infection can induce an abnormal IgA immune response like Henoch-Schönlein purpura, quite similar to typical acute post-infectious glomerulonephritis. Indeed, hypocomplementemia that is typical of acute glomerulonephritis has also been described in Henoch-Schönlein purpura.</p> <p>Case presentation</p> <p>We describe a 14-year-old Caucasian Spanish girl who developed urinary abnormalities and cutaneous purpura after streptococcal infection. Renal biopsy showed typical findings from Henoch-Schönlein purpura nephritis. In addition, she had low serum levels of complement (C4 fraction) that persisted during follow-up, in spite of her clinical evolution. She responded to treatment with enalapril and steroids.</p> <p>Conclusion</p> <p>The case described has, at least, three points of interest in Henoch-Schönlein purpura: 1) Initial presentation was preceded by streptococcal infection; 2) There was a persistence of low serum levels of complement; and 3) There was response to steroids and angiotensin-converting enzyme inhibitor in the presence of nephrotic syndrome. There are not many cases described in the literature with these characteristics. We conclude that Henoch-Schönlein purpura could appear after streptococcal infection in patients with abnormal complement levels, and that steroids and angiotensin-converting enzyme inhibitor could be successful treatment for the disease.</p

Brain age predicts long-term recovery in post-stroke aphasia

Funding Information: This study was supported by the following grant sponsors: National Institute on Deafness and Other Communication Disorders (P50 DC014664; DC008355); National Institute of Neurological Disorders and Stroke (NS054266). Publisher Copyright: © 2022 The Author(s).The association between age and language recovery in stroke remains unclear. Here, we used neuroimaging data to estimate brain age, a measure of structural integrity, and examined the extent to which brain age at stroke onset is associated with (i) cross-sectional language performance, and (ii) longitudinal recovery of language function, beyond chronological age alone. A total of 49 participants (age: 65.2 ± 12.2 years, 25 female) underwent routine clinical neuroimaging (T1) and a bedside evaluation of language performance (Bedside Evaluation Screening Test-2) at onset of left hemisphere stroke. Brain age was estimated from enantiomorphically reconstructed brain scans using a machine learning algorithm trained on a large sample of healthy adults. A subsample of 30 participants returned for follow-up language assessments at least 2 years after stroke onset. To account for variability in age at stroke, we calculated proportional brain age difference, i.e. the proportional difference between brain age and chronological age. Multiple regression models were constructed to test the effects of proportional brain age difference on language outcomes. Lesion volume and chronological age were included as covariates in all models. Accelerated brain age compared with age was associated with worse overall aphasia severity (F(1, 48) = 5.65, P = 0.022), naming (F(1, 48) = 5.13, P = 0.028), and speech repetition (F(1, 48) = 8.49, P = 0.006) at stroke onset. Follow-up assessments were carried out ≥2 years after onset; decelerated brain age relative to age was significantly associated with reduced overall aphasia severity (F(1, 26) = 5.45, P = 0.028) and marginally failed to reach statistical significance for auditory comprehension (F(1, 26) = 2.87, P = 0.103). Proportional brain age difference was not found to be associated with changes in naming (F(1, 26) = 0.23, P = 0.880) and speech repetition (F(1, 26) = 0.00, P = 0.978). Chronological age was only associated with naming performance at stroke onset (F(1, 48) = 4.18, P = 0.047). These results indicate that brain age as estimated based on routine clinical brain scans may be a strong biomarker for language function and recovery after stroke.Peer reviewe

Presentations of children to emergency departments across Europe and the COVID-19 pandemic: A multinational observational study

BACKGROUND: During the initial phase of the Coronavirus Disease 2019 (COVID-19) pandemic, reduced numbers of acutely ill or injured children presented to emergency departments (EDs). Concerns were raised about the potential for delayed and more severe presentations and an increase in diagnoses such as diabetic ketoacidosis and mental health issues. This multinational observational study aimed to study the number of children presenting to EDs across Europe during the early COVID-19 pandemic and factors influencing this and to investigate changes in severity of illness and diagnoses. METHODS AND FINDINGS: Routine health data were extracted retrospectively from electronic patient records of children aged 18 years and under, presenting to 38 EDs in 16 European countries for the period January 2018 to May 2020, using predefined and standardized data domains. Observed and predicted numbers of ED attendances were calculated for the period February 2020 to May 2020. Poisson models and incidence rate ratios (IRRs), using predicted counts for each site as offset to adjust for case-mix differences, were used to compare age groups, diagnoses, and outcomes. Reductions in pediatric ED attendances, hospital admissions, and high triage urgencies were seen in all participating sites. ED attendances were relatively higher in countries with lower SARS-CoV-2 prevalence (IRR 2.26, 95% CI 1.90 to 2.70, p < 0.001) and in children aged <12 months (12 to <24 months IRR 0.86, 95% CI 0.84 to 0.89; 2 to <5 years IRR 0.80, 95% CI 0.78 to 0.82; 5 to <12 years IRR 0.68, 95% CI 0.67 to 0.70; 12 to 18 years IRR 0.72, 95% CI 0.70 to 0.74; versus age <12 months as reference group, p < 0.001). The lowering of pediatric intensive care admissions was not as great as that of general admissions (IRR 1.30, 95% CI 1.16 to 1.45, p < 0.001). Lower triage urgencies were reduced more than higher triage urgencies (urgent triage IRR 1.10, 95% CI 1.08 to 1.12; emergent and very urgent triage IRR 1.53, 95% CI 1.49 to 1.57; versus nonurgent triage category, p < 0.001). Reductions were highest and sustained throughout the study period for children with communicable infectious diseases. The main limitation was the retrospective nature of the study, using routine clinical data from a wide range of European hospitals and health systems. CONCLUSIONS: Reductions in ED attendances were seen across Europe during the first COVID-19 lockdown period. More severely ill children continued to attend hospital more frequently compared to those with minor injuries and illnesses, although absolute numbers fell. TRIAL REGISTRATION: ISRCTN91495258 https://www.isrctn.com/ISRCTN91495258

Pathogenesis of Henoch-Schönlein purpura nephritis

The severity of renal involvement is the major factor determining the long-term outcome of children with Henoch-Schönlein purpura (HSP) nephritis (HSPN). Approximately 40% children with HSP develop nephritis, usually within 4 to 6 weeks after the initial onset of the typical purpuric rashes. Although the pathogenetic mechanisms are still not fully delineated, several studies suggest that galactose-deficient IgA1 (Gd-IgA1) is recognized by anti-glycan antibodies, leading to the formation of the circulating immune complexes and their mesangial deposition that induce renal injury in HSPN

Holocene Cyclic Records of Ice-Rafted Debris and Sea Ice Variations on the East Greenland and Northwest Iceland Margins

The dynamics of the Greenland Ice Sheet and drift of sea ice from the Arctic Ocean reaching Denmark Strait are poorly constrained. We present data on the provenance of Fe oxide detrital grains from two cores in the Denmark Strait area and compare the Fe grain source data with other environmental proxies in order to document the variations and potential periodicities in ice-rafted debris delivery during the Holocene. Based on their Fe grain geochemistry, the sediments can be traced to East Greenland sources and to more distal sites around the Arctic Basin. On the Holocene time scales of the two cores, sea ice biomarker (IP25) data, and quartz weight percent reveal positive associations with T°C and inverse associations with biogenic carbonate wt%. Trends in the data were obtained from Singular Spectrum Analysis (SSA), and residuals were tested for cyclicity. Trends on the environmental proxies explained between 15 and 90% of the variance. At both sites the primary Fe grain sources were from Greenland, but significant contributions were also noted from Banks Island and Svalbard. There is a prominent cyclicity of 800 yrs as well as other less prominent cycles for both Greenland and arctic sources. The Fe grain sources from Greenland and the circum-Arctic Ocean are in synchronization, suggesting that the forcings for these cycles are regional and not local ice sheet instabilities

Boðkerfi í æðaþeli tengd histamín og thrombín miðlaðri NO-myndun. Hlutverk AMP-örvaðs prótein kínasa

Healthy, normally functioning endothelium is critical for maintenance of vascular homeostasis. Endothelial dysfunction, on the other hand, is a key early step in numerous vascular diseases. A vascular surface that normally is thromboresistant, antiinflammatory, vasodilatory and antiproliferative can turn into a surface that is thrombogenic, proinflammatory, vasoconstrictive and stimulatory of smooth muscle cell proliferation. In certain disease states such as atherosclerosis, hypertension and diabetes such changes may involve a chronically perturbed vascular behaviour critical for disease progression. The signaling that mediates the response to environmental changes has in recent years attracted attention. The transduction of these signals is critical for an appropriate cellular response while derailment of the signaling can be a key feature of a pathologic response or disease. One of the crucial responses of the vascular endothelium to a variety of signals is the production of NO from arginine through the activation of eNOS. The main goal of this work was to enhance our understanding of the biological and physiological properties of the healthy vascular endothelium with regards to the activity of eNOS. Specifically, we sought to elucidate the signaling pathway involved in mediating endothelial stimulation by the G-protein linked agonists thrombin and histamine with emphasis on the role of AMPK in maintaining normal NO-production. We demonstrated the presence of a PI3K-Akt independent pathway activating eNOS in HUVEC after treatment of cells with histamine or thrombin. We showed that both agonists inhibited Akt phosphorylation by activation of PKCδ and that the phosphorylation of eNOS at Ser1177 by these agonists was unaffected by the PI3K-inhibitor wortmannin. After having ruled out several kinases known to phosphorylate eNOS we hypothesized a role for AMPK based on preliminary results showing AMPK phosphorylation by histamine and thrombin. We were able to demonstrate a role for AMPK in this previously unrecognised pathway, making AMPK an important link in mediating thrombin and histamine induced activation of eNOS independent of PI3K-Akt. AMPK was phosphorylated by these agonists and activation was demonstrated by phosphorylation of ACC a known target of AMPK. eNOS phosphorylation and activation after histamine or thrombin was inhibited by H89, an inhibitor of AMPK and PKA. Also, AICAR and CCCP, both known to activate AMPK, caused phosphorylation of eNOS. The Ca+2-chelator BAPTA inhibited the phosphorylation of AMPK, eNOS and NO-formation, demonstrating the Ca+2-dependency of this pathway. With LKB1 being the only kinase known to activate AMPK in mammals at this time, we suggested an LKB1-AMPK dependent pathway causing eNOS activation after treatment with histamine or thrombin, possibly explained by activation of energy-consuming pathways by the elevation of intracellular Ca+2. We discovered that this pathway was only activated under culture conditions that allowed a sharp but brief fall in cellular ATP after thrombin or histamine stimulation. Under culture conditions that prevented or did not allow such a fall in the energy level of the endothelial cells AMPK was still activated but exclusively via a different upstream pathway and downstream this AMPK-activation played no role in the activation of eNOS. Also, we demonstrated, using siRNA, that the AMPK-eNOS pathway is dependent on the activation of AMPK through LKB1. Downregulation of the α2-isoform of AMPK but not α1 resulted in shrinkage of the cells and loss of contact between cells suggesting that the α2-isoform of AMPK is critical for endothelial integrity. The inhibiting effects of the SOD-mimetic Tempol on these morphologic changes seen in AMPKα2-downregulated cells and the increased expression of the ER stress marker GRP78 caused by this downregulation also suggested a role for AMPKα2 in oxidant defences. We confirm a pathway dependent on CaMKK causing activation of eNOS under culture conditions where intracellular ATP is unchanged without the involvement of (LKB1-)AMPK, as also described by others. However, we also demonstrate a previously unknown pathway, activated by a rise in intracellular AMP/ATP ratio, causing LKB1-AMPK-dependent phosphorylation and activation of eNOS by thrombin or histamine. This pathway is activated under culture conditions where the intracellular AMP/ATP ratio is elevated by these agonists. Our fundamental discovery that environmental conditions dictate which pathway is activated is interesting but even more so the key role played by the ATP-level, a fall in the cellular energy level being a well known consequence of pathologic conditions such as ischemia, hypoxia and infection. Although thrombin can activate eNOS through an AMPK-independent pathway the activation of the LKB1-AMPK pathway greatly enhances the NO-production and may therefore be of major importance for endothelial function and endothelial health.The Research Fund of the University of Iceland, the Research Fund of Landspitali-University Hospital, the Research Fund of the Counsil for Science and Technology, the Helga Jónsdottir and Sigurliði Kristjánsson Memorial Fund and the Eimskipafelag University Fund