Bacteriology of aspiration pneumonia in patients with acute coma.
Loss of protective airway reflexes in patients with acute coma puts these patients at risk of aspiration pneumonia complicating the course of the primary disease. Available data vary considerably with regard to bacteriology, role of anaerobic bacteria, and antibiotic treatment. Our objective was to research the bacteriology of aspiration pneumonia in acute coma patients who were not pre-treated with antibiotics or hospitalized within 30 days prior to the event. We prospectively analyzed 127 patient records from adult patients admitted, intubated and ventilated to a tertiary medical intensive care unit with acute coma. Bacteriology and antibiotic resistance testing from tracheal aspirate sampled within 24 h after admission, blood cultures, ICU scores (APACHE II, SOFA), hematology, and clinical chemistry were assessed. Patients were followed up until death or hospital discharge. The majority of patients with acute coma suffered from acute cardiovascular disorders, predominantly myocardial infarction, followed by poisonings, and coma of unknown cause. In a majority of our patients, microaspiration resulted in overt infection. Most frequently S. aureus, H. influenzae, and S. pneumoniae were isolated. Anaerobic bacteria (Bacteroides spec., Fusobacteria, Prevotella spec.) were isolated from tracheal aspirate in a minority of patients, and predominantly as part of a mixed infection. Antibiotic monotherapy with a 2nd generation cephalosporin, or a 3rd generation gyrase inhibitor, was most effective in our patients regardless of the presence of anaerobic bacteria.


[A metastatic muscle mass as the first manifestation of renal-cell cancer. A case report and review of the literature].
Although metastasis is relatively frequent in renal cell carcinoma (affecting mainly the lungs, lymphatic glands and bones), location of metastasis in muscles is extremely rare as in other oncological processes. The literature only offers isolated cases of renal cell carcinoma with metastasis in muscles, and no preferred muscular site is found in these cases nor is there any clear intrinsic relation to prognosis, having been described both at the onset an upon relapse of the oncological process. The intimate mechanism behind this infrequent metastatic site is unknown even today but it presents a series of specific and peculiar factors such as vascular flow, metabolism and oxygen tissue pressure which can negatively condition this process. We present the case of a 60-year-old patient who began with a mass in the triceps muscle. A biopsy confirmed its metastatic nature and indicated an asymptomatic renal cell carcinoma. Further complementary examinations showed metastasis in other muscle groups.


Large-scale evaluation of log P predictors: local corrections may compensate insufficient accuracy and need of experimentally testing every other compound.
A large variety of log P calculation methods failed to produce sufficient accuracy in log P prediction for two in-house datasets of more than 96000 compounds contrary to their significantly better performances on public datasets. The minimum Root Mean Squared Error (RMSE) of 1.02 and 0.65 were calculated for the Pfizer and Nycomed datasets, respectively, in the 'out-of-box' implementation. Importantly, the use of local corrections (LC) implemented in the ALOGPS program based on experimental in-house log P data significantly reduced the RMSE to 0.59 and 0.48 for the Pfizer and Nycomed datasets, respectively, instantly without retraining the model. Moreover, more than 60% of molecules predicted with the highest confidence in each set had a mean absolute error (MAE) less than 0.33 log units that is only ca. 10% higher than the estimated variation in experimental log P measurements for the Pfizer dataset. Therefore, following this retrospective analysis, we suggest that the use of the predicted log P values with high confidence may eliminate the need of experimentally testing every other compound. This strategy could reduce the cost of measurements for pharmaceutical companies by a factor of 2, increase the confidence in prediction at the analog design stage of drug discovery programs, and could be extended to other ADMET properties.


Characterization of the complex formed by β-glucocerebrosidase and the lysosomal integral membrane protein type-2.
The lysosomal integral membrane protein type-2 (LIMP-2) plays a pivotal role in the delivery of β-glucocerebrosidase (GC) to lysosomes. Mutations in GC result in Gaucher's disease (GD) and are the major genetic risk factor for the development of Parkinson's disease (PD). Variants in the LIMP-2 gene cause action myoclonus-renal failure syndrome and also have been linked to PD. Given the importance of GC and LIMP-2 in disease pathogenesis, we studied their interaction sites in more detail. Our previous data demonstrated that the crystal structure of LIMP-2 displays a hydrophobic three-helix bundle composed of helices 4, 5, and 7, of which helix 5 and 7 are important for ligand binding. Here, we identified a similar helical motif in GC through surface potential analysis. Coimmunoprecipitation and immunofluorescence studies revealed a triple-helical interface region within GC as critical for LIMP-2 binding and lysosomal transport. Based on these findings, we generated a LIMP-2 helix 5-derived peptide that precipitated and activated recombinant wild-type and GD-associated N370S mutant GC in vitro. The helix 5 peptide fused to a cell-penetrating peptide also activated endogenous lysosomal GC and reduced α-synuclein levels, suggesting that LIMP-2-derived peptides can be used to activate endogenous as well as recombinant wild-type or mutant GC efficiently. Our data also provide a structural model of the LIMP-2/GC complex that will facilitate the development of GC chaperones and activators as potential therapeutics for GD, PD, and related synucleinopathies.


Stochastic fluctuations in apoptotic threshold of tumour cells can enhance apoptosis and combat fractional killing.
Fractional killing, which is a significant impediment to successful chemotherapy, is observed even in a population of genetically identical cancer cells exposed to apoptosis-inducing agents. This phenomenon arises not from genetic mutation but from cell-to-cell variation in the activation timing and level of the proteins that regulates apoptosis. To understand the mechanism behind the phenomenon, we formulate complex fractional killing processes as a first-passage time (FPT) problem with a stochastically fluctuating boundary. Analytical calculations are performed for the FPT distribution in a toy model of stochastic p53 gene expression, where the cancer cell is killed only when the p53 expression level crosses an active apoptotic threshold. Counterintuitively, we find that threshold fluctuations can effectively enhance cellular killing by significantly decreasing the mean time that the p53 protein reaches the threshold level for the first time. Moreover, faster fluctuations lead to the killing of more cells. These qualitative results imply that fluctuations in threshold are a non-negligible stochastic source, and can be taken as a strategy for combating fractional killing of cancer cells.


A Two-Stage Flow Cytometry Strategy to Distinguish Single Cells from Doublets in Heterogeneous Cell Mixtures and Improve Cell Cluster Identification: Application to Human Monocyte Subpopulations.
Flow cytometry is a powerful method, widely used to identify cell types present in tissues, to describe their phenotypes, and to purify cells for functional analyses. As a single cell technique, flow cytometry relies on identifying and excluding cell doublets and aggregates present in samples in the initial gating steps. This identification is based on detection of events generating electrical pulses falling outside of linear variations of pulse height, width, and area in a singlet population with increasing cell sizes. In heterogeneous cell mixtures, however, with cell types varying extensively in size and granularity, exclusion of doublets has the risk of removing single cells that co-localize with doublets of another cell type. This is particularly the case when doublets of a smaller cell type overlap with large cells of a distinct, larger cell type. Here, we describe a gating method to reduce this risk. In this protocol, initial gating steps aim to segregate cells according to physical characteristics (such as size and granularity) and gene expression properties in order to obtain more homogeneous cell clusters. Doublet exclusion is then performed separately in each cluster, minimizing the risk of confusion between single cells and doublets. To illustrate this protocol, human blood monocytes are separated and analyzed. By implementing this protocol, we were able to reveal the existence of a population of large monocytes previously unrecognized using conventional gating strategies. In subsequent functional assays, we have shown that this novel population exhibits unique inflammatory responses, highlighting the need and pertinence of this approach to identify and characterize infrequent-yet functionally relevant-cell populations present in complex cell mixtures. © 2021 Wiley Periodicals LLC. Basic Protocol: Distinguishing single cells from doublets in heterogeneous cell mixtures by flow cytometry.


Adjunct Screening With Tomosynthesis or Ultrasound in Women With Mammography-Negative Dense Breasts: Interim Report of a Prospective Comparative Trial.
Debate on adjunct screening in women with dense breasts has followed legislation requiring that women be informed about their mammographic density and related adjunct imaging. Ultrasound or tomosynthesis can detect breast cancer (BC) in mammography-negative dense breasts, but these modalities have not been directly compared in prospective trials. We conducted a trial of adjunct screening to compare, within the same participants, incremental BC detection by tomosynthesis and ultrasound in mammography-negative dense breasts. Adjunct Screening With Tomosynthesis or Ultrasound in Women With Mammography-Negative Dense Breasts is a prospective multicenter study recruiting asymptomatic women with mammography-negative screens and dense breasts. Eligible women had tomosynthesis and physician-performed ultrasound with independent interpretation of adjunct imaging. Outcome measures included cancer detection rate (CDR), number of false-positive (FP) recalls, and incremental CDR for each modality; these were compared using McNemar's test for paired binary data in a preplanned interim analysis. Among 3,231 mammography-negative screening participants (median age, 51 years; interquartile range, 44 to 78 years) with dense breasts, 24 additional BCs were detected (23 invasive): 13 tomosynthesis-detected BCs (incremental CDR, 4.0 per 1,000 screens; 95% CI, 1.8 to 6.2) versus 23 ultrasound-detected BCs (incremental CDR, 7.1 per 1,000 screens; 95% CI, 4.2 to 10.0), P = .006. Incremental FP recall occurred in 107 participants (3.33%; 95% CI, 2.72% to 3.96%). FP recall (any testing) did not differ between tomosynthesis (FP = 53) and ultrasound (FP = 65), P = .26; FP recall (biopsy) also did not differ between tomosynthesis (FP = 22) and ultrasound (FP = 24), P = .86. The Adjunct Screening With Tomosynthesis or Ultrasound in Women With Mammography-Negative Dense Breasts' interim analysis shows that ultrasound has better incremental BC detection than tomosynthesis in mammography-negative dense breasts at a similar FP-recall rate. However, future application of adjunct screening should consider that tomosynthesis detected more than 50% of the additional BCs in these women and could potentially be the primary screening modality.


Species-specific Microorganisms in Acid-tolerant Chironomus Larvae Reared in a Neutral pH Range under Laboratory Conditions: Single Dataset Analysis.
To obtain a more detailed understanding of organismal acid tolerance, the larval microbiomes of 11 Chironomus species collected from acidic or neutral pH areas in Japan and reared at pH 7-8 under laboratory conditions were systematically compared using an amplicon sequencing ana-lysis. Evenness values were lower for the larval microbiomes of acid-tolerant Chironomus cf. riparius, Chironomus fusciceps, and Chironomus sulfurosus than for eight acid-sensitive species based on an alpha diversity ana-lysis. The lower evenness observed suggested a biased abundance of microorganisms, which was consistent with the identification of Chironomus species-specific microorganisms (such as Agromyces mediolanus and Comamonas odontotermitis related bacteria) with high abundance in acid-tolerant larvae. The abundance of specific microorganisms was also high in the microbiome of acid-tolerant larvae of Chironomus acerbiphilus reared at pH 4, but not in that of acid-sensitive larvae. Based on a PICRUSt2 ana-lysis, genes involved in saccharide transport were less abundant in the microbiome of acid-tolerant larvae than in that of acid-sensitive larvae, indicating nutrient-poor acidic environments. Although these results were obtained from single datasets, acid-tolerant larvae appeared to establish Chironomus species-specific interactions with microorganisms independent of saccharides, in contrast to acid-sensitive larvae. The present study is the first step towards understanding organismal acid tolerance.


Reducing Peak Energy Demand among Residents Who Are Not Billed for Their Electricity Consumption: Experimental Evaluation of Behaviour Change Interventions in a University Setting.
Behaviour change interventions aiming to reduce household energy consumption are regarded as an effective means to address disparities between demand and supply and reduce emissions. Less recognised is their success in shifting consumers' energy consumption from peak demand periods to off-peak times of the day. This study reports two experiments that test the effect of feedback and reminder notifications on energy consumption in university halls-of-residence. A quasi-experiment and a randomised controlled experiment were conducted with residential students to evaluate behaviour change interventions aimed at reducing daily peak and critical peak demand, respectively. The results of Experiment One (n = 143) demonstrated significant reductions in the energy use of the treatment group relative to the control. On average, the treatment group's energy use was 12.4 per cent lower than their pre-intervention baseline. In Experiment Two (n = 88), normative elements of the intervention were supplemented with a reminder notification prior to the onset of the simulated critical peak demand period. The results showed that, relative to the control condition, the 8-h notification reduced demand by 20% on average with a 12% decrease for the 24-h notification (with 2-h follow-up). These results indicate that peak energy issues can be alleviated using low-cost and easily implemented behaviour change strategies.


Speech acquisition within a 3-generation Vietnamese-English family: the influence of maturation and ambient phonology.
The emergence approach to speech acquisition theorises the influence of intrinsic capabilities (e.g., maturation), interactional capabilities, and extrinsic contexts (e.g., ambient phonology). Intrinsic and extrinsic influences were examined via a case study of a 3-generation Vietnamese-English family with two brothers (C1 aged 5;6 and C2 aged 3;10), their mother (M), grandfather (GF) and grandmother (GM). Their speech was assessed using the Diagnostic Evaluation of Articulation and Phonology (DEAP) and the Vietnamese Speech Assessment (VSA). Standard Australian English/Standard Vietnamese productions were defined as 'correct', even though the adults spoke different Vietnamese dialects. Their percentage of standard consonants correct (PSCC) was: C1 (English:92.27%, Vietnamese:89.05%), C2 (E:86.47%, VN:86.13%), M (E:90.34%, VN:96.35%), GF (E:82.61%, VN:97.81%), GM (VN:99.27%). Percentages were higher when dialectal variants were included. C1 and C2 had more pronunciation matches with English (86.96%) than Vietnamese (79.56%). C1's pronunciation matched: M (E:85.02%, VN:83.94%), GF (E:79.23%, VN:77.37%), GM (VN:73.72%) and C2's pronunciation matched: M (E:79.23%, VN:73.72%), GF (E:73.91%, VN:75.18%), GM (VN:72.26%). There was evidence of ambient phonology influences and cross-linguistic transfer. For example, in Vietnamese 'r' is produced as /ʐ/ or /r/ , but was produced by C1 as [ɹ] (English approximant) and by C2 [w] (age-appropriate/ɹ/substitution). The children demonstrated maturation influences for late-occurring English consonants (e.g., English /θ/ →[f]). This study found evidence for the emergence approach and recommends knowledge of the ambient phonology augments traditional child-focused understandings of children's speech acquisition.


Design of an ultrasound-guided robotic brachytherapy needle-insertion system.
In this paper we describe a new robotic brachytherapy needle-insertion system that is designed to replace the template used in the manual technique. After a brief review of existing robotic systems, we describe the requirements that we based our design upon. A detailed description of the proposed system follows. Our design is capable of positioning and inclining a needle within the same workspace as the manual template. To help improve accuracy, the needle can be rotated about its axis during insertion into the prostate. The system can be mounted on existing steppers and also easily accommodates existing seed dispensers, such as the Mick Applicator.


Shared Representational Formats for Information Maintained in Working Memory and Information Retrieved from Long-Term Memory.
Current theories propose that the short-term retention of information in working memory (WM) and the recall of information from long-term memory (LTM) are supported by overlapping neural mechanisms in occipital and parietal cortex. However, the extent of the shared representations between WM and LTM is unclear. We designed a spatial memory task that allowed us to directly compare the representations of remembered spatial information in WM and LTM with carefully matched behavioral response precision between tasks. Using multivariate pattern analyses on functional magnetic resonance imaging data, we show that visual memories were represented in a sensory-like code in both memory tasks across retinotopic regions in occipital and parietal cortex. Regions in lateral parietal cortex also encoded remembered locations in both tasks, but in a format that differed from sensory-evoked activity. These results suggest a striking correspondence in the format of representations maintained in WM and retrieved from LTM across occipital and parietal cortex. On the other hand, we also show that activity patterns in nearly all parietal regions, but not occipital regions, contained information that could discriminate between WM and LTM trials. Our data provide new evidence for theories of memory systems and the representation of mnemonic content.


A synergistic effect of albumin and fibrinogen on immunoglobulin-induced red blood cell aggregation.
Therapeutic administration of immunoglobulins (Ig) has the potential to precipitate thrombotic events. This phenomenon may be explained by red blood cell (RBC) aggregation, which can be potentiated by Ig. The contribution of plasma albumin and fibrinogen to Ig-induced RBC aggregation is unclear. We examined RBC aggregation in three settings: 1) patients receiving therapeutic infusions of Ig; 2) patients receiving plasma supplemented in vitro with Ig; and 3) patients receiving RBC suspensions in standard buffer with varying concentrations of albumin, Ig, and fibrinogen. Ig infusion augmented aggregation of RBCs from patients with normal or high plasma levels of albumin but decreased aggregation in those with lower plasma albumin concentrations. In vitro, RBC aggregation was significantly increased only when all three components, fibrinogen, albumin, and Ig, were present at or above normal concentrations in the suspension but was unaffected when any one of the components was absent from the suspension. Our results suggest a three-way interaction among fibrinogen, Ig, and albumin that synergistically induces RBC aggregation in plasma. Understanding these interactions may help predict clinically important phenomena related to RBC aggregation, such as thrombotic complications of Ig infusion.


Deletion of GSK-3beta in mice leads to hypertrophic cardiomyopathy secondary to cardiomyoblast hyperproliferation.
Based on extensive preclinical data, glycogen synthase kinase-3 (GSK-3) has been proposed to be a viable drug target for a wide variety of disease states, ranging from diabetes to bipolar disorder. Since these new drugs, which will be more powerful GSK-3 inhibitors than lithium, may potentially be given to women of childbearing potential, and since it has controversially been suggested that lithium therapy might be linked to congenital cardiac defects, we asked whether GSK-3 family members are required for normal heart development in mice. We report that terminal cardiomyocyte differentiation was substantially blunted in Gsk3b(-/-) embryoid bodies. While GSK-3alpha-deficient mice were born without a cardiac phenotype, no live-born Gsk3b(-/-) pups were recovered. The Gsk3b(-/-) embryos had a double outlet RV, ventricular septal defects, and hypertrophic myopathy, with near obliteration of the ventricular cavities. The hypertrophic myopathy was caused by cardiomyocyte hyperproliferation without hypertrophy and was associated with increased expression and nuclear localization of three regulators of proliferation - GATA4, cyclin D1, and c-Myc. These studies, which we believe are the first in mammals to examine the role of GSK-3alpha and GSK-3beta in the heart using loss-of-function approaches, implicate GSK-3beta as a central regulator of embryonic cardiomyocyte proliferation and differentiation, as well as of outflow tract development. Although controversy over the teratogenic effects of lithium remains, our studies suggest that caution should be exercised in the use of newer, more potent drugs targeting GSK-3 in women of childbearing age.


Low levels of Her2/neu expressed by Ewing's family tumor cell lines can redirect cytokine-induced killer cells.
To identify novel treatments for pediatric solid tumors and/or for malignancies with low-level Her2/neu expression. Using fluorescence-activated cell sorting and immunohistochemistry, Her2/neu expression was determined on cell lines derived vfrom Ewing's family tumors (EFT) and neuroblastoma. Sensitivity to trastuzumab treatment was investigated using an in vitro proliferation assay. Cytotoxicity against EFT cell lines was done with either freshly isolated or ex vivo activated and expanded T cells (cytokine-induced killer cells, CIK cells), with or without addition of a CD3xHer2/neu bispecific antibody. The effects of either trastuzumab, CIK cells alone, or CD3xHer2/neu bispecific antibody redirected CIK cells was determined using a SCID/hu model of EFTs and serial, noninvasive bioluminescent imaging. EFT cell lines express 5- to 10-fold lower levels of her2/neu than either breast (BT-474) or ovarian (SK-OV-3) cell lines. Treatment of EFT cell lines with trastuzumab did not induce growth inhibition either in vitro or in vivo. In contrast, Her2/neu could be used to redirect CIK cell to mediate cytotoxicity against EFTs both in vitro and in vivo (using two different treatment schemas). CD3xHer2/neu bispecific antibody and CIK cells may be a suitable approach to treat malignancies with low-level Her2/neu expression not responsive to trastuzumab.


Evaluation of Plaque Morphology by 64-Slice Coronary Computed Tomographic Angiography Compared to Intravascular Ultrasound in Nonocclusive Segments of Coronary Arteries.
Although intravascular ultrasound (IVUS) is the current gold standard for plaque characterization, noninvasive coronary computed tomographic angiography (CCTA) requires further evaluation. The ability to detect plaque morphology by CCTA remains unclear. The purpose of this study was to evaluate the diagnostic accuracy of CCTA for plaque detection and morphology. Thirty-one patients underwent cardiac catheterization with IVUS and CCTA. The presence of plaque was evaluated by both modalities in nonocclusive segments (<50% stenosis) of the left anterior descending artery, left circumflex artery, and right coronary artery. Plaque morphology was classified as (1) normal, (2) soft or fibrous, (3) fibrocalcific, or (4) calcific. Results by IVUS and CCTA were compared blindly on a segment-to-segment basis with subgroup analysis based on CCTA tube voltage. Among the 31 patients (mean age 56.2 ± 8.6 years, 27% female), 152 segments were analyzed. Of these segments, 42% were in the left anterior descending artery, 32% were in the left circumflex artery, and 26% were in the right coronary artery. Plaque morphology by IVUS identified 103 segments as fibrous (68%), 31 as fibrocalcific (20%), and 6 as calcific (4.0%); 12 segments were normal (8.0%). To evaluate for the presence of plaque, CCTA had an overall sensitivity and specificity of 99% and 75%, respectively. In patients who underwent CCTA with a tube voltage of 100 kV, both sensitivity and specificity were 100%. The sensitivity and specificity of CCTA to identify plaque as calcified (fibrocalcific or calcific) vs noncalcified (soft or fibrous) were 87% and 96%, respectively. Overall, the accuracy of CCTA to detect the presence of plaque was 97%; the accuracy to detect plaque calcification was 94%. CCTA offers excellent sensitivity and accuracy for plaque detection and morphology characterization in nonocclusive coronary segments. In addition, diagnostic accuracy is preserved with a reduced tube voltage protocol.


Presence of double-stranded RNAs in human and calf isolates of Cryptosporidium parvum.
We examined the occurrence of 2 virus-like double-stranded (ds)RNAs in human and calf isolates of Cryptosporidium parvum senso latu and other microorganisms, including 7 other members of the genus. A total of 32 isolates of C. parium, 16 from humans (5 from acquired immune deficiency syndrome patients) and 16 from calves, were analyzed. Ethidium bromide staining, or Northern blot analysis, or reverse transcription/polymerase chain reaction, or all 3 methods, revealed that both genotype 1 and genotype 2 isolates of C. parvum possessed these dsRNAs. No other Cryptosporidium spp. or other organisms examined possessed these dsRNAs. Comparison analysis of partial cDNA sequences of dsRNAs from human and calf isolates revealed a high degree of similarity (>92% and >93% identical nucleotides for large and small dsRNAs, respectively). Slight, consistent differences in nucleotide sequences could be seen at select sites and were associated with an isolate being either genotype 1 or 2. Because of the widespread distribution of the dsRNAs, the similarity of these molecules between isolates, and high host specificity, these nucleic acids may prove to represent species-specific molecular markers for C. parvum. Evidence also suggests that the dsRNA can be utilized for molecular genotyping of C. parvum.


Adjuvant heparanase inhibitor PI-88 therapy for hepatocellular carcinoma recurrence.
To demonstrate that administering heparanase inhibitor PI-88 at 160 mg/d is safe and promising in reducing hepatocellular carcinoma (HCC) recurrence for up to 3 year following curative resection. A total of 143 patients (83.1% of the 172 participants in the phase II study) participated in the follow-up study. Of these patients, 50 had received no treatment, 48 had received 160 mg/d PI-88, and 45 had received 250 mg/d PI-88 during the phase II trial. Safety parameters and the following efficacy endpoints were investigated: (1) time to recurrence; (2) disease-free survival; and (3) overall survival. PI-88 at 160 mg/d delayed the onset and frequency of HCC recurrence, and provided a clinically significant survival advantage for up to 3 years after treatment compared with those of the control group: (1) the recurrence-free rate increased from 50% to 63%, and (2) time to recurrence at the 36th percentile was postponed by 78%. The efficacy of administering PI-88 at 250 mg/d was confounded by a high dropout rate (11 out of 54 patients). Additionally, subgroup analyses of patients with (1) multiple tumors or a single tumor ≥ 2 cm; and (2) hepatitis B or C revealed that administering PI-88 at 160 mg/d conferred the most significant survival advantage (56.8% improvement in disease-free survival, P = 0.045) for patients with both risk factors for recurrence. Administering PI-88 at 160 mg/d is a safe and well-tolerated dosage that may confer significant clinical benefits for patients with HCC.


MR imaging of diabetic foot infection.
Infection of the foot is common, especially in the diabetic population. Over the past decade, magnetic resonance imaging (MRI) has become a 'gold standard' for evaluation of pedal infection. Therefore it is important for the radiologist to understand the MRI appearance of various manifestations of infection as well as conditions that may simulate infection, including neuropathic disease.


Feeding and oviposition rates in the pine weevil Hylobius abietis (Coleoptera: Curculionidae).
Feeding and oviposition in the pine weevil Hylobius abietis (Linnaeus) were monitored under laboratory conditions in two long-term experiments lasting over an extended breeding season. Data were also collected from weevils under semi-natural conditions outdoors. In addition, the effects of crowding and starvation were studied in separate experiments. During the main peak oviposition period, female H. abietis consumed 50% more bark tissue than males. When oviposition ceased, the feeding rate of the females declined to the same level as in the males. The rates and spatial distribution patterns of oviposition and feeding were clearly affected by climatic conditions and the degree of crowding. Females were estimated to lay on average 0.8 eggs per day during the season under outdoor conditions. The realized fecundity of a female weevil during the first season was estimated to be approximately 70 eggs. The estimated average rate of feeding was 23 mm2 of Scots pine bark per weevil per day. This implies that planted seedlings can only constitute a minor part of the food resources needed to sustain H. abietispopulations of the size that usually appear on fresh clear-cuttings in northern Europe.


Allelism of MCKD, FJHN and GCKD caused by impairment of uromodulin export dynamics.
The disease complex medullary cystic disease/familial juvenile hyperuricemic nephropathy (MCKD/FJHN) is characterized by alteration of urinary concentrating ability, frequent hyperuricemia, tubulo-interstitial fibrosis, cysts at the cortico-medullary junction and renal failure. MCKD/FJHN is caused by mutations of the gene encoding uromodulin, the most abundant protein in urine. Here, we describe new missense mutations in three families with MCKD/FJHN and demonstrate allelism with a glomerulocystic kidney disease (GCKD) variant, showing association of cyst dilatation and collapse of glomeruli with some clinical features similar to MCKD/FJHN as hyperuricemia and impairment of urine concentrating ability. Furthermore, we provide the first functional characterization of uromodulin mutations. The four newly identified mutants were characterized by immunofluorescence and FACS analysis on transfected cells. These experiments showed that all uromodulin mutations cause a delay in protein export to the plasma membrane due to a longer retention time in the endoplasmic reticulum. Immunohistochemistry on GCKD and MCKD/FJHN kidney biopsies revealed dense intracellular accumulation of uromodulin in tubular epithelia of the thick ascending limb of Henle's loop. Electron microscopy demonstrated accumulation of dense fibrillar material within the endoplasmic reticulum. Consistently, patient urines show a severe reduction of excreted uromodulin. The maturation impairment is consistent with the clinical findings and suggests a pathogenetic mechanism leading to these kidney diseases.


Declining length of hospital stay for pneumonia and postdischarge outcomes.
This study was designed to assess 8-year trends in the duration of hospitalization for community-acquired pneumonia and to evaluate the impact of declining length of stay on postdischarge short-term readmission and mortality. We conducted a prospective observational cohort study of 1886 patients with community-acquired pneumonia who were discharged from a single hospital between March 1, 2000, and June 30, 2007. The main outcomes measured were all-cause mortality and hospital readmission during the 30-day period after discharge. Regression models were used to identify risk factors associated with hospital length of stay and the adjusted associations between length of stay and mortality and readmission. Factors associated with a longer hospital stay included the number of comorbid conditions, high risk classification on the Pneumonia Severity Index, bilateral or multilobe radiographic involvement, and treatment failure. Patients treated with an appropriate antibiotic were less likely to have an increased length of stay. The mean length of stay was significantly shorter during the 2006 to 2007 period (3.6 days) than during the 2000 to 2001 period (5.6 days, P<.001). Despite the reduction in length of stay, there were no significant differences in the likelihood of death or readmission at 30 days between the 2 time periods. Adjusted multivariate analysis showed that patients with hospital stays less than 3 days did not have significant increases in postdischarge outcomes. The marked decreased in the length of stay for patients hospitalized with community-acquired pneumonia since 2000 has not been accompanied by an increase in short-term mortality or hospital readmission.


Differential detectability of polymorphic warning signals under varying light environments.
The striking colour-pattern variation of some aposematic species is paradoxical because selection by predators is expected to favour signal uniformity. Although the mechanisms allowing for the maintenance of such variation are not well understood, possible explanations include both non-adaptive processes like drift and gene flow; and adaptive processes, such as an interaction between natural and sexual selection, spatial and temporal variation in selection, a link between behaviour or other fitness-related traits and phenotype, and predators' ability to generalise among different signals. Here we test whether warning-signal polymorphisms, such as that of dyeing poison frogs (Dendrobates tinctorius), could be maintained by differences in detectability among morphs. We did experiments in the wild using wax models with different aposematic colour patterns vs. cryptic ones, and examined the attack rates by wild predators over time. We also tested the detectability of different aposematic morphs by 'human predators' under different light environments. We found that cryptic frog models were attacked more than aposematic models, but there were no differences in bird attack rates towards the different aposematic morphs. However, we found that detectability of different morphs depends both on predator experience and light environment. We suggest that the interaction between differential detectability and signal efficiency among morphs in different light conditions could be a mechanism aiding to the maintenance of warning-signal polymorphisms. Our results highlight the importance of considering the light environment at which predators have their first encounters with aposematic prey for future studies on predation in the wild.


Successful combined lung and kidney transplantation for pulmonary lymphangioleiomyomatosis and renal angiolipomas.
Pulmonary lymphangioleiomyomatosis (LAM) and renal angiolipomas are rare but distinct clinical entities that share similar morphological features. Lung transplantation is considered as a valuable therapeutic modality in patients with end-stage pulmonary LAM. However, in some patients, renal complications due to bleeding angiomyolipomas and cyclosporin-induced nephropathy have become newly identified problems. This study reports the first case of combined lung and kidney transplantation for pulmonary lymphangioleiomyomatosis and renal angiolipomas. Two years after transplantation, renal and pulmonary function have remained stable and the patient has resumed a normal daily life, including a full-time professional activity.


Targeting Causes of Burnout in Residency: An Innovative Approach Used at Hennepin Healthcare.
Rates of burnout are high in physicians in the United States. While others have reported on the success of burnout-reduction strategies on practicing physicians and residents, few strategies have approached the problem longitudinally in residents. From 2014 to 2019, the authors used a previously developed survey to assess factors related to resident burnout, including sleep, personal time, professional fulfillment, effects on relationships, program recognition, and peer support. At Hennepin Healthcare, a safety-net hospital in Minneapolis, Minnesota, the authors created a reproducible process for collecting data from internal medicine residents annually, and for using evidence-based conceptual frameworks to develop a continuous improvement method to address worklife across training years. Interventions included jeopardy coverage for essential life events, a newsletter celebrating resident achievements, removal of after-hours consult pager call, an extra day off for senior residents on the wards, and care packages distributed to night teams. Annually from 2014 to 2019, 40/66 (60.6%) to 62/73 residents (84.9%) completed the survey (average response rate was 72.1% over 6 years). Survey results were shared with residents in multiple formats, and feedback was requested, demonstrating that burnout reduction is a priority for program leadership. High professional fulfillment scores were documented every year. Self-reported rates of burnout were between 25% and 35%. Significant improvements were seen in perception of empathy, sleep impairment, and peer support. The authors developed a plan for minimizing burnout, which includes the following evidence-based domains: workload, control, balance in effort and reward, work-life balance, fairness, values, support, gender equity, moral distress, and moral injury. Additional interventions include protected time for didactics, trauma-informed care training, and addressing workplace racism. The authors aspire to achieve an integrated culture of well-being for residents and faculty; foster an efficient, effective, and fair learning environment; and reduce-and ultimately eliminate-burnout.


Understanding the reasons why Chinese older people do not wish to tell their life stories.
To explore the reasons why older Chinese people decline to tell their life stories. Promoting the telling of life stories by older people, as part of healthy ageing, may develop their sense of unity and meaning in life. However, not all older adults seem to review their past and tell others about it. Understanding is lacking about how 'healthy' older adults experience the meanings of not telling. A descriptive phenomenology with Giorgi's analysis. Data were collected from June 2010-August 2011 through unstructured individual interviews. A potential sample of 36 older people were identified through convenience sampling from 17 daycare and senior centres, of whom 19 participated in this study. Our findings suggest that the reasons why these older people declined to tell their life stories relate to an understanding of the self and to the perception that the older people had of their audience's response and their family's support. Older people seemed to be naturally inclined to reminiscence, but their accumulated personal and interpersonal experiences might have caused them to feel disinclined to talk about what they no longer considered to be important. An understanding about the tension existing in cultural/generational narratives of the older people may help health professionals to provide them with a supportive environment for their continuous construction of social identities. This may contribute to their better social health. Equally important is to provide an opportunity for them to share their stories and engage in family and cross-generational dialogues.


Treatment with dilute alkali-nuclease S1 permits the analysis of DNA damage: cells treated with platinum analogues.
We describe here an approach to characterize various lesions induced in DNA by drug treatments, using three parameters: (a) release of single-stranded DNA fragments by cell lysis in dilute alkali, which result from enzymatic strand scission during DNA repair or chemical alterations of DNA; (b) the presence of high molecular weight DNA in cells after lysis in dilute alkali followed by nuclease S1 treatment which, due to drug-induced DNA cross-links and its level is a measure of the amount of DNA-containing cross-links; and (c) the appearance of small double-stranded DNA fragments, when the cell lysis is followed by digestion with nuclease S1 to remove single-stranded DNA. This DNA shows the same characteristics as DNA of untreated cells, but it may contain monoadducts. By following the flow of label through the three parameters, one can characterize both the lesions induced in DNA and how the lesions are repaired. We report here results of three platinum analogues: cis-Pt(II), trans-Pt(II), and cis-FLAP(II). A large proportion of DNA in treated cells appears as fragments (parameter c). The cis- compounds and trans- compounds differ with regard to appearance of high molecular weight DNA (parameter b) and the initial release of fragments (parameter a).


[Usefulness of melatonin in the diagnostics and therapy of pineal gland and brain neoplasms].
Melatonin or N-acetyl-5-methoxytryptamine is mainly produced by pineal gland and exhibits characteristic diurnal rhythm of synthesis and secretion with the highest levels at night and low levels during a day. For several years the knowledge of pineal gland and its hormone was rather poor. It was last years when knowledge of the role of melatonin in health and different diseases was developed. Nowadays we are aware that melatonin plays important role in diagnosis and therapy of several diseases, and may be helpful factor also in neurosurgery. Author presents the review of the present knowledge on the role of melatonin in diagnosis of pineal region pathology and treatment of brain tumors.


Severity assessment of acute pulmonary embolism with spiral CT: evaluation of two modified angiographic scores and comparison with clinical data.
Spiral computed tomography (CT) has shown promising results in the detection of acute pulmonary embolism. The aim of this study was to investigate whether the severity of acute pulmonary embolism could be quantitatively assessed with spiral CT examinations and to test the potential clinical impact of this information. In a consecutive series of 123 patients screened with spiral CT for suspected acute pulmonary embolism, 31 patients (25%) had evidence of emboli. The severity of pulmonary arterial obstruction in those 31 spiral CT examinations was evaluated by two independent observers using angiographic scores previously described by Walsh (29) and Miller (30), adapted to the needs of spiral CT. Clinical patient subgroups were defined according to oxygen saturation, heart rate, and echocardiographic signs of right ventricular strain. CT severity scores were then correlated to each other and to clinical parameters using the Spearman rank test. Interobserver agreement was calculated using the analysis of variance. Both modified Walsh and Miller scores were readily reproducible and showed interobserver agreements of 0.85 and 0.96, respectively (p = 0.001). Patients with mild and marked clinical abnormalities showed statistically significant differences between CT severity scores. Differences between severity scores of patients with moderate and marked clinical abnormalities were somewhat significant. No significant mean severity score differences were seen between patients with mild and moderate clinical abnormalities. Although correlations of severity scores and detailed clinical parameters within the defined subgroups were moderate to poor, threshold scores greater than 10 (Miller) and greater than 11 (Walsh) always indicated marked clinical abnormalities. The modified scores presented in this study constitute a readily reproducible method for the quantitative assessment of acute pulmonary embolism severity on spiral CT examinations.


Evolution of coreceptor use and CD4-independence in envelope clones derived from SIVsm-infected macaques.
Coreceptor use of HIV can evolve during infection. We previously examined coreceptor use of related SIVsm inoculum viruses and sequential reisolates from cynomolgus macaques. These viruses exhibited broad coreceptor specificities and, generally, CCR5 use remained efficient and stable, while alternative coreceptor use decreased longitudinally. Here we demonstrate that individual envelopes (Envs) from inoculum and reisolate viruses fuse via a range of coreceptors, including CCR5, CCR8, CXCR6, GPR15, GPR1, and APJ. On the whole, coreceptor use of Envs from sequential reisolates recapitulated that of reisolate viruses, thus CCR5 use remained stable while alternative coreceptor use tended to decrease over time. Rhesus CCR5, GPR15, and CXCR6 supported fusion to a similar extent as their human counterparts. Additionally, a number of Envs mediated CD4-independent fusion via CCR5 and GPR15. Envs from different inoculum viruses exhibited distinct dependencies on CD4 for fusion via CCR5, ranging from strictly CD4-dependent to efficiently CD4-independent. Early reisolates from macaques infected with CD4-independent inoculums maintained or evolved Envs with a broad range of CD4-independence. CD4-independence became less variable/efficient in late reisolates from macaques that developed neutralizing antibodies. Infection with a CD4-dependent virus resulted in evolution of CD4-independent Envs in late reisolates. While CD4 independence can potentially broaden tropism in vivo, CD4-independent viruses are particularly sensitive to neutralizing antibodies. Therefore, interplay between receptor tropism and neutralization may shape viral evolution and SIV pathogenesis.


Disinfection of wastewater by hydrogen peroxide or peracetic acid: development of procedures for measurement of residual disinfectant and application to a physicochemically treated municipal effluent.
The Montreal Urban Community Wastewater Treatment Plant (MUCWTP) located in Montreal. Quebec, Canada, uses physicochemical treatment processes prior to discharging wastewater into the St. Lawrence River via an outfall tunnel of 2 hours retention time. Although chlorination facilities exist, they are not being used, and the MUCWTP is seeking alternative methods for disinfection to achieve a 2- to 3-log fecal coliform reduction. Liquid chemical disinfectants were attractive options because of their low capital costs. This led to an investigation of the feasibility of using hydrogen peroxide or peracetic acid. A method for measuring peroxycompounds (hydrogen peroxide or peracetic acid plus hydrogen peroxide) was developed using the peroxidase-based oxidation of 2,2'-azino-bis(3-ethylbenz-thiazoline-6-sulfuric acid) diammonium salt (ABTS) with hydrogen peroxide. The validity of the method was confirmed using effluent from the MUCWTP. Recovery was higher than 90% for peracetic acid levels as low as 1.0 mg/L. Quenching of hydrogen peroxide was achieved with 50-mg/L catalase; quenching of peracetic acid was achieved with 100 mg/L of sodium thiosulfate, followed by 50 mg/L of catalase. Batch disinfection tests were conducted on MUCWTP effluent. Hydrogen peroxide and peracetic acid in wastewater over time could be modeled as a second-order decay, with the decay "constant" being a function of the initial concentration of peroxycompounds. This function was the same for both hydrogen peroxide and peracetic acid, possibly indicating similar decomposition pathways in wastewater matrices. Disinfection was modeled using a modified Hom equation. Required doses of hydrogen peroxide to reach the target fecal coliform levels ranged from 106 to 285 mg/L, with the higher doses occurring when ferric chloride instead of alum was used as the coagulant. Hence, hydrogen peroxide was infeasible as a disinfectant for this application. On the other hand, the peracetic acid dose needed to achieve the target fecal coliform level was only 0.6 to 1.6 mg/L. Therefore, peracetic acid seems to be a promising disinfectant for physicochemical or primary effluent, or combined sewer overflows.


Histological changes in the epiphyseal plate of growing young adult male rats after long-term hydrocortisone and salmon calcitonin administration.
Glucocorticosteroids decrease the longitudinal growth of bones. It has been reported that skeletally immature rats on calcitonin treatment grow longer and slimmer. The aim of this study was to assess whether concomitant treatment with calcitonin would influence longitudinal growth and histology of epiphyseal plate in hydrocortisone-treated rats. Fifty six growing male Wistar rats were randomly divided into two control and four experimental groups. Rats in the first two experimental groups were treated i.p. b.i.d. with hydrocortisone, while to the other experimental animals salmon calcitonin s.c. was administered concomitantly. The controls received appropriate vehicula. The rats were kept under standardised conditions. The first phase of the experiment was completed after 28 days and right femora and tibiae were harvested from groups and preserved in 4% buffered formalin. Lengths of the fresh femora were measured with an electronic caliper. After preservation, tibial bones were decalcified in 5% EDTA at room temperature. Paraffin-embedded coronal slices 10 μm thick from proximal tibia were taken and stained with hematoxylin/ eosin and alcian blue for histological assessment of the epiphyseal plate. A similar procedure was repeated after 56 days in the second phase of the experiment. The femora of hydrocortisone-treated rats were significantly shorter than those of controls, but after concomitant administration of salmon calcitonin the difference was insignificant. The histological assessment showed that depressed function of growth plate in hydrocortisone-treated animals after both phases of the experiment was partially reversed by salmon calcitonin treatment. Prolonged concomitant treatment with high-doses of salmon calcitonin partially reverses glucocorticoid-induced depression of rat growth plate.


Modification of epigenetic patterns in low birth weight children: importance of hypomethylation of the ACE gene promoter.
There is a growing body of evidence that epigenetic alterations are involved in the pathological mechanisms of many chronic disorders linked to fetal programming. Angiotensin-converting enzyme (ACE) appears as one candidate gene that brings new insights into the epigenetic control and later development of diseases. In this view, we have postulated that epigenetic modifications in the ACE gene might show different interactions between birth weight (BW), blood pressure levels, plasma ACE activity and ACE I/D polymorphism. To explore this hypothesis, we performed a cross-sectional study to evaluate the DNA methylation of 3 CpG sites using pyrosequencing within the ACE gene promoter of peripheral blood leukocytes from 45 LBW children compared with 70 NBW children. Our results have revealed that LBW children have lower methylation levels (P<0.001) in parallel with a higher ACE activity (P = 0.001). Adjusting for prematurity, gender, age, body mass index, and family history of cardiovascular disease did not alter these findings. We have also performed analyses of individual CpG sites. The frequency of DNA methylation was significantly different at two CpG sites (site 1: nucleotide position +555; and site 3: nucleotide position +563). In addition, we have found a significant inverse correlation between degree of DNA methylation and both ACE activity (P<0.001) and systolic blood pressure levels (P<0.001). We also observed that the methylation level was significantly lower in LBW children who are carriers of the DD genotype compared to NBW children with DD genotype (P<0.024). In conclusion, we are able to demonstrate that the hypomethylation in the 3 CpG sites of ACE gene promoter is associated with LBW in 6 to 12 year-old children. The magnitude of these epigenetic changes appears to be clinically important, which is supported by the observation that discrete changes in DNA methylation can affect systolic blood pressure and ACE protein activity levels.


Experimental characterization and modelization of ion exchange kinetics for a carboxylic resin in infinite solution volume conditions. Application to monovalent-trivalent cations exchange.
This study is devoted to the characterization of ion exchange inside a microsphere of carboxylic resin. It aims at describing the kinetics of this exchange reaction which is known to be controlled by interdiffusion in the particle. The fractional attainment of equilibrium function of time depends on the concentration of the cations in the resin which can be modelized by the Nernst-Planck equation. A powerful approach for the numerical resolution of this equation is introduced in this paper. This modeling is based on the work of Helfferich but involves an implicit numerical scheme which reduces the computational cost. Knowing the diffusion coefficients of the cations in the resin and the radius of the spherical exchanger, the kinetics can be hence completely determined. When those diffusion parameters are missing, they can be deduced by fitting experimental data of fractional attainment of equilibrium. An efficient optimization tool coupled with the implicit resolution has been developed for this purpose. A monovalent/trivalent cation exchange had been experimentally characterized for a carboxylic resin. Diffusion coefficients and concentration profiles in the resin were then deduced through this new model.


Cluster-Based Approach Utilizing Optimally Tuned TD-DFT to Calculate Absorption Spectra of Organic Semiconductor Thin Films.
The photophysics of organic semiconductor (OSC) thin films or crystals has garnered significant attention in recent years since a comprehensive theoretical understanding of the various processes occurring upon photoexcitation is crucial for assessing the efficiency of OSC materials. To date, research in this area has relied on methods using Frenkel-Holstein Hamiltonians, calculations of the GW-Bethe-Salpeter equation with periodic boundaries, or cluster-based approaches using quantum chemical methods, with each of the three approaches having distinct advantages and disadvantages. In this work, we introduce an optimally tuned, range-separated time-dependent density functional theory approach to accurately reproduce the total and polarization-resolved absorption spectra of pentacene, tetracene, and perylene thin films, all representative OSC materials. Our approach achieves excellent agreement with experimental data (mostly ≤0.1 eV) when combined with the utilization of clusters comprising multiple monomers and a standard polarizable continuum model to simulate the thin-film environment. Our protocol therefore addresses a major drawback of cluster-based approaches and makes them attractive tools for OSC investigations. Its key advantages include its independence from external, system-specific fitting parameters and its straightforward application with well-known quantum chemical program codes. It demonstrates how chemical intuition can help to reduce computational cost and still arrive at chemically meaningful and almost quantitative results.


Risperidone induced ventricular tachycardia.
This is a case report of ventricular tachycardia in a 35 years old African American male being treated with risperidone for schizophrenia. Patient had no other cardiovascular risk factors. His physical examination and laboratory test results were essentially normal. Episodes of non-sustained ventricular tachycardia resolved after discontinuation of risperidone. Above case report is likely to add to the limited evidence for risperidone associated cardiotoxicity.


Context-Dependent Modulation of Early Visual Cortical Responses to Numerical and Nonnumerical Magnitudes.
Whether and how the brain encodes discrete numerical magnitude differently from continuous nonnumerical magnitude is hotly debated. In a previous set of studies, we orthogonally varied numerical (numerosity) and nonnumerical (size and spacing) dimensions of dot arrays and demonstrated a strong modulation of early visual evoked potentials (VEPs) by numerosity and not by nonnumerical dimensions. Although very little is known about the brain's response to systematic changes in continuous dimensions of a dot array, some authors intuit that the visual processing stream must be more sensitive to continuous magnitude information than to numerosity. To address this possibility, we measured VEPs of participants viewing dot arrays that changed exclusively in one nonnumerical magnitude dimension at a time (size or spacing) while holding numerosity constant and compared this to a condition where numerosity was changed while holding size and spacing constant. We found reliable but small neural sensitivity to exclusive changes in size and spacing; however, exclusively changing numerosity elicited a much more robust modulation of the VEPs. Together with previous work, these findings suggest that sensitivity to magnitude dimensions in early visual cortex is context dependent: The brain is moderately sensitive to changes in size and spacing when numerosity is held constant, but sensitivity to these continuous variables diminishes to a negligible level when numerosity is allowed to vary at the same time. Neurophysiological explanations for the encoding and context dependency of numerical and nonnumerical magnitudes are proposed within the framework of neuronal normalization.


Robust, Universal Tree Balance Indices.
Balance indices that quantify the symmetry of branching events and the compactness of trees are widely used to compare evolutionary processes or tree-generating algorithms. Yet, existing indices are not defined for all rooted trees, are unreliable for comparing trees with different numbers of leaves, and are sensitive to the presence or absence of rare types. The contributions of this article are twofold. First, we define a new class of robust, universal tree balance indices. These indices take a form similar to Colless' index but can account for population sizes, are defined for trees with any degree distribution, and enable meaningful comparison of trees with different numbers of leaves. Second, we show that for bifurcating and all other full m-ary cladograms (in which every internal node has the same out-degree), one such Colless-like index is equivalent to the normalized reciprocal of Sackin's index. Hence, we both unify and generalize the two most popular existing tree balance indices. Our indices are intrinsically normalized and can be computed in linear time. We conclude that these more widely applicable indices have the potential to supersede those in current use. [Cancer; clone tree; Colless index; Sackin index; species tree; tree balance.].


Longitudinal epigenetic variation of DNA methyltransferase genes is associated with vulnerability to post-traumatic stress disorder.
Epigenetic differences exist between trauma-exposed individuals with and without post-traumatic stress disorder (PTSD). It is unclear whether these epigenetic differences pre-exist, or arise following, trauma and PTSD onset. In pre- and post-trauma samples from a subset of Detroit Neighborhood Health Study participants, DNA methylation (DNAm) was measured at DNA methyltransferase 1 (DNMT1), DNMT3A, DNMT3B and DNMT3L. Pre-trauma DNAm differences and changes in DNAm from pre- to post-trauma were assessed between and within PTSD cases (n = 30) and age-, gender- and trauma exposure-matched controls (n = 30). Pre-trauma DNAm was tested for association with post-trauma symptom severity (PTSS) change. Potential functional consequences of DNAm differences were explored via bioinformatic search for putative transcription factor binding sites (TFBS). DNMT1 DNAm increased following trauma in PTSD cases (p = 0.001), but not controls (p = 0.067). DNMT3A and DNMT3B DNAm increased following trauma in both cases (DNMT3A: p = 0.009; DNMT3B: p < 0.001) and controls (DNMT3A: p = 0.002; DNMT3B: p < 0.001). In cases only, pre-trauma DNAm was lower at a DNMT3B CpG site that overlaps with a TFBS involved in epigenetic regulation (p = 0.001); lower pre-trauma DNMT3B DNAm at this site was predictive of worsening of PTSS post-trauma (p = 0.034). Some effects were attenuated following correction for multiple hypothesis testing. DNAm among trauma-exposed individuals shows both longitudinal changes and pre-existing epigenetic states that differentiate individuals who are resilient versus susceptible to PTSD. These distinctive DNAm differences within DNMT loci may contribute to genome-wide epigenetic profiles of PTSD.


Assessment of antibiotic release and antibacterial efficacy from pendant glutathione hydrogels using ex vivo porcine skin.
Acute bacterial skin and skin structure infections (ABSSSIs) confer a substantial burden on the healthcare system. Local antibiotic delivery systems can provide controlled drug release directly to the site of infection to maximize efficacy and minimize systemic toxicity. The purpose of this study was to examine the antibacterial activity of antibiotic-loaded glutathione-conjugated PEG hydrogels (GSH-PEG) against ABSSSIs utilizing an ex vivo porcine dermal explant model. Vancomycin- or meropenem-loaded GSH-PEG hydrogels at 3 different dose levels were loaded over 1 h. Drug release was monitored in vitro under submerged conditions, by the Franz cell diffusion method, and ex vivo utilizing a porcine dermis model. Antibacterial activity was assessed ex vivo on porcine dermis explants inoculated with Staphylococcus aureus or Pseudomonas aeruginosa isolates treated with vancomycin- or meropenem-loaded GSH-PEG hydrogels, respectively. Histological assessment of the explants was conducted to evaluate tissue integrity and viability in the context of the experimental conditions. A dose-dependent release was observed from vancomycin and meropenem hydrogels, with in vitro Franz cell diffusion data closely representing ex vivo vancomycin release, but not high dose meropenem release. High dose vancomycin-loaded hydrogels resulted in a >3 log10 clearance against all S. aureus isolates at 48 h. High dose meropenem-loaded hydrogels achieved 6.5, 4, and 2 log10 reductions in CFU/ml against susceptible, intermediate, and resistant P. aeruginosa isolates, respectively. Our findings demonstrate the potential application of GSH-PEG hydrogels for flexible, local antibiotic delivery against bacterial skin infections.


Code blue: Predictors of survival.
Code blue is a rapid response system developed for emergency resuscitation and stabilization of any sudden cardiac arrest (SCA) within a hospital. Literatures on outcome and factors predicting mortality from SCA in the Emergency departments (EDs) of India is scant. This retrospective cohort study included all patients above the age of 15 years who had a code blue declared in the ED between the months of January 2018 and June 2019. Factors related to the sustained return of spontaneous circulation (ROSC) and mortality were analyzed using descriptive-analytic statistics and logistic regressions. This study included 435 patients with a male predominance of 299 (69%). The mean age was 54.5 (SD - 16.5) years. Resuscitation was not attempted for 18 patients because of the terminal nature of the underlying disease. The majority were in-hospital cardiac arrests (74%). The nonshockable rhythm included pulseless electrical activity (PEA) (85.5%) and asystole (14.5%) cases. Shockable rhythms, that is, pulseless ventricular tachycardia/ventricular fibrillation were noted in only 10% (43/417) of cases. ROSC was attained in 184 (44.1%) patients, among which 56 (13.4%) were discharged alive from the hospital. Multivariate logistic regression analysis showed CPR >10 min (odds ratio [OR]: 13.58; 95% CI: 8.39-22.01; P < 0.001) and female gender (OR: 1.89; 95% CI: 1.13-3.17; P = 0.016) to be independent risk factors for failure to achieve ROSC in ED. The initial documented rhythm was nonshockable in the majority of the cases. CPR duration of more than 10 min and female gender were independent risk factors for failure to achieve ROSC in the ED. Nonshockable rhythms have a poorer outcomes than that of shockable rhythms.


Structural and biological characterization of bovine insulin-like growth factor binding protein-3.
Insulin-like growth factor binding protein-3 (IGFBP-3) purified from bovine serum shares 19 of 25 amino-terminal amino acid residues with IGFBP-3 purified from human, rat, and porcine sources. A newly characterized bovine fibroblast model was used to investigate the biological effects of purified bovine IGFBP-3 (bIGFBP-3). Coincubation of insulin-like growth factor I (IGF-I) with increasing concentrations of bIGFBP-3 produced a dose-dependent inhibition of IGF-I-stimulated [3H]aminoisobutyric acid (AIB) uptake in cultured bovine fibroblasts. Inhibition was complete at equimolar concentrations of IGF-I and bIGFBP-3. Inhibition of IGF-I-stimulated [3H]AIB uptake paralleled the ability of bIGFBP-3 to prevent [125I]IGF-I cell surface binding. In contrast, preincubation with bIGFBP-3 resulted in a dose-dependent enhancement of IGF-I-stimulated [3H]AIB uptake; a 32-86% increase in IGF-I bioactivity was seen after a 24 h preexposure to 10 nM bIGFBP-3, and a 2- to 6-fold potentiation was seen after a 72 h preincubation. Preincubation with bIGFBP-3 increased both the sensitivity and maximal responsiveness of the cells to IGF-I. The potentiating effects of bIGFBP-3 were associated with increased [125I]IGF-I binding to cultured bovine fibroblasts. Affinity cross-linking experiments indicated that the increase in IGF-I binding was due to increased membrane-associated bIGFBP-3 rather than to a bIGFBP-3-induced increase in type I IGF receptors. bIGFBP-3 had no effect on insulin stimulation of [3H]AIB uptake under either experimental condition. These data suggest that soluble bIGFBP-3 inhibits IGF-I action by sequestering and preventing IGF-I receptor binding, whereas surface-associated bIGFBP-3 enhances the growth-promoting effects of IGF-I in bovine fibroblasts. We propose that IGFBP-3 serves a dual function in modulating IGF action in vivo.


Dietary calcium and pregnancy-induced hypertension: is there a relation?
The evidence that calcium plays a role in the etiology, prevention, and treatment of pregnancy-induced hypertension (PIH) is reviewed. The precise factors involved in the pathogenesis of PIH are unclear, but several alterations in calcium metabolism have been identified. Epidemiologic data suggest an inverse correlation between dietary calcium intake and incidence of PIH. Although evidence suggests a possible beneficial effect of supplemental calcium, contradictions persist in clinical trials of pregnant women. Presently, there is insufficient evidence to support routine calcium supplementation of all pregnant women. However, high-risk groups, such as pregnant teens, populations with inadequate calcium intake, and women at risk of developing PIH, may benefit from consuming additional dietary calcium.


Canine scent detection in the diagnosis of lung cancer: revisiting a puzzling phenomenon.
Patient prognosis in lung cancer largely depends on early diagnosis. The exhaled breath of patients may represent the ideal specimen for future lung cancer screening. However, the clinical applicability of current diagnostic sensor technologies based on signal pattern analysis remains incalculable due to their inability to identify a clear target. To test the robustness of the presence of a so far unknown volatile organic compound in the breath of patients with lung cancer, sniffer dogs were applied. Exhalation samples of 220 volunteers (healthy individuals, confirmed lung cancer or chronic obstructive pulmonary disease (COPD)) were presented to sniffer dogs following a rigid scientific protocol. Patient history, drug administration and clinicopathological data were analysed to identify potential bias or confounders. Lung cancer was identified with an overall sensitivity of 71% and a specificity of 93%. Lung cancer detection was independent from COPD and the presence of tobacco smoke and food odours. Logistic regression identified two drugs as potential confounders. It must be assumed that a robust and specific volatile organic compound (or pattern) is present in the breath of patients with lung cancer. Additional research efforts are required to overcome the current technical limitations of electronic sensor technologies to engineer a clinically applicable screening tool.


Electronic communications and home blood pressure monitoring (e-BP) study: design, delivery, and evaluation framework.
Randomized controlled trials have provided unequivocal evidence that treatment of hypertension decreases mortality and major disability from cardiovascular disease; however, blood pressure remains inadequately treated in most affected individuals. This large gap continues despite the facts that more than 90% of adults with hypertension have health insurance, and hypertension is the leading cause of visits to the doctor. New approaches are needed to improve hypertension care. The Electronic Communications and Home Blood Pressure Monitoring (e-BP) study is a three-arm randomized controlled trial designed to determine whether care based on the Chronic Care Model and delivered over the Internet improves hypertension care. The primary study outcomes are systolic, diastolic, and blood pressure control; secondary outcomes are medication adherence, patient self-efficacy, satisfaction and quality of life, and healthcare utilization and costs. Hypertensive patients receiving care at Group Health medical centers are eligible if they have uncontrolled blood pressure on two screening visits and access to the Web and an e-mail address. Study participants are randomly assigned to three intervention groups: (a) usual care; (b) home blood pressure monitoring receipt and proficiency training on its use and the Group Health secure patient website (with secure e-mail access to their healthcare provider, access to a shared medical record, prescription refill and other services); or (c) this plus pharmacist care management (collaborative care management between the patient, the pharmacist, and the patient's physician via a secure patient website and the electronic medical record). We will determine whether a new model of patient-centered care that leverages Web communications, self-monitoring, and collaborative care management improves hypertension control. If this model proves successful and cost-effective, similar interventions could be used to improve the care of large numbers of patients with uncontrolled hypertension.


Immune-mediated disorders associated with idiopathic sudden sensorineural hearing loss.
Different immune disorders are involved in the development of sudden sensorineural hearing loss (SSNHL). This includes a wide spectrum of immune-mediated disorders such as immune complexes, production of autoantibodies to the inner ear proteins, production of anticardiolipin (aCL) antibodies, and cellular immune defects. Some studies have also found an elevation of total IgE levels and a type 1 immune reaction. Our objective was to establish the association of "idiopathic" SSNHL (ISSNHL) with various autoantibodies, and to analyze the persistence over time of existing aCL and anti-beta2 glycoprotein 1 (anti-beta2 GP1) antibodies in such patients. Finally, we sought to establish a possible association between ISSNHL and total IgE elevation and whether this elevation is due to a specific type I immune reaction. In this prospective follow-up study, 51 patients considered as having ISSNHL were compared with 35 age-sex matched healthy volunteers over a 3-year period. All participants were tested for serum antinuclear antibodies, antithyroid antibodies, aCL, and rheumatoid factor. All patients who were positive for aCL antibodies were reanalyzed 3 months later for aCL antibody persistence and for the coexistence and persistence of anti-beta2 GP1 antibodies. Skin prick tests were performed in patients in whom total IgE was elevated. Antinuclear antibodies were positive in 9 of 51 (17%) and antithyroid antibodies in 11 of 51 (21%) ISSNHL patients, as compared to 1 of 35 (3%) and 2 of 35 (6%), respectively, in the control group. Rheumatoid factor was positive in 6 of the 51 patients (12%) and in none of the control group. Positive aCL antibodies were present in 16 of 51 patients (31%), 6 of whom (12%) were also positive for anti-beta2 GP1 antibodies. Three months later, aCL antibodies persisted in 7 patients (14%), and anti-beta2 GP1 in 4 patients. Only 2 of the normal subjects (6%) were positive for aCL antibodies, which persisted in only 1 of them (3%). The level of total IgE was elevated in 14 of 51 patients (27%), in contrast to 3 of 35 controls (8%). Six of them (42%) demonstrated a positive skin test to at least 1 allergen, but only 3 presented allergy symptoms. Our findings support the reported existence of multiple immune-mediated disorders in patients with ISSNHL. The lack of aCL antibody persistence in as many as half of our patients strongly suggests that transient phenomena (eg, viral infection) may trigger aCL antibody activity. However, the presence of aCL antibodies, especially in conjunction with anti-beta2 GP1 antibodies, suggests that in some patients, SSNHL is included among the rare symptoms of the antiphospholipid syndrome.


Superfund Cleanups and Children's Lead Exposure.
This study evaluates the effect of EPA's Superfund cleanup program on children's lead exposure. We linked two decades of blood lead level (BLL) measurements from children in six states with data on Superfund sites and other lead risk factors. We used quasi-experimental methods to identify the causal effect of proximity to Superfund cleanups on rates of elevated BLL. We estimated a difference-in-difference model comparing the change in elevated BLL of children closer to versus farther from lead-contaminated sites before, during, and after cleanup. We also estimated a triple difference model including children near hazardous sites with minimal to no lead contamination as a comparison group. We used spatial fixed effects and matching to minimize potential bias from unobserved differences between the treatment and comparison groups. Results indicate that Superfund cleanups lowered the risk of elevated BLL for children living within 2 kilometers of lead-contaminated sites 13 to 26 percent.


Preoperative Predictors of Free Flap Failure.
Microvascular free tissue transfer is an important reconstructive option for defects of the head and neck. The present study aims to identify preoperative patient- and tumor-specific characteristics, laboratory values, and other risk factors associated with early free flap failure. Retrospective cohort study of 1070 patients. Head and neck surgical oncology service at a tertiary care center from 2005 to 2019. Demographics, comorbidities, preoperative laboratory values, treatment history, and cancer stage were collected for consecutive patients who underwent free flap reconstruction of the head and neck and experienced early free flap failure (<8 days from surgery). In 1070 patients, the prevalence of early free flap failure was 3.8% (n = 41). Female sex (odds ratio [OR], 2.58; 95% CI, 1.36-4.99), presence of peripheral vascular disease (OR, 2.78; 95% CI, 1.05-6.57), and elevated preoperative platelet count (OR, 2.67; 95% CI, 1.20-5.47) were independently associated with risk of early free flap failure. Female sex, peripheral vascular disease, and preoperative thrombocytosis are all strong predictors of early free flap failure. This suggests that hypercoagulability and poor vessel quality may predispose patients to flap loss. Patients with elevated platelets or peripheral vascular disease warrant careful reconstructive decision making and close monitoring in the perioperative period.


Isolation and characterization human chorion membrane trophoblast and mesenchymal cells.
To develop protocols for isolation and culture of human chorionic mesenchymal and trophoblast cells and test their differential responsiveness to oxidative stress. Chorion trophoblast cells (CTC) and chorion mesenchymal cells (CMC) were isolated from term fetal membranes by modifying current protocols. Their purity and characteristics were tested using bright field microscopy and after staining for cytokeratin (CK)-7 and vimentin. Cigarette smoke extract (CSE) was used to stimulate cells, and we determined reactive oxygen species (ROS) production using 2'7'-dichlorodihydro-fluorescein assay, stress signaler p38MAPK activation (Western blot) and senescence by flow cytometry. Co-treatment with antioxidant N-acetyl cystine (NAC) either alone or in combination with SB203580 (p38MAPK inhibitor) was used to test oxidative stress (OS)- and p38MAPK-mediated effects. The isolation and cell culture protocol used in this study yielded 92% pure CTC and 100% pure CMC. CSE treatment significantly induced ROS production, P-p38MAPK activation, and senescence in both cell types compared to controls. Cotreatment with NAC reduced ROS production and p38MAPK activation, and co-treatment with both NAC and SB203580 reduced senescence. ROS response in CMC was higher than CTC; however, senescence of CTC was 10-fold higher than CMC. We introduce approaches for proper isolation and culture of CTC and CMC without any influence or overgrowth of one specific type cell that can confound results. Using this approach, we determined differential effects of CTC and CMC to OS condition seen at term labor. Both CTC and CMC undergo p38MAPK-mediated senescence; however, the rate of senescence is higher in CTC.


Cryptococcosis in liver and kidney transplant recipients receiving anti-thymocyte globulin or alemtuzumab.
Rabbit anti-thymocyte globulin (ATG) and alemtuzumab have been used for induction or preconditioning and for the treatment of acute rejection in organ transplant recipients in many centers. Such regimens may lead to a substantial decline in the CD4 lymphocyte count to levels seen in other population groups at high risk of cryptococcosis. In view of this, we examined the impact of such therapy on the cumulative incidence of cryptococcosis among liver and kidney recipients. A total of 834 liver and 727 kidney transplants were performed during the study period. Seven hundred and eighty-one patients did not receive ATG or alemtuzumab; 646 received 1 dose of either drug, and 134 patients received 2 doses of either drug. The cumulative incidence of cryptococcosis was 0.26% (2/781) among those who did not receive ATG or alemtuzumab; 0.3% (2/646) among those who received only 1 dose, and 2.24% (3/134) among those who received 2 doses (P=0.03). There were 5 cases of cryptococcosis in liver recipients and 2 in kidney recipients. There were 3 cases of cryptococcal meningitis, 3 of pneumonia, and 1 of disseminated disease. The 2 kidney recipients had meningitis. Diagnosis occurred at a median of 255 days (range 7-517) after transplantation. The mortality rate was 14.2%. We conclude that the use of 1 dose of ATG or alemtuzumab is not associated with an increased cumulative incidence of cryptococcosis, but that those patients receiving 2 doses are at increased risk.


Physically inspired deep learning of molecular excitations and photoemission spectra.
Modern functional materials consist of large molecular building blocks with significant chemical complexity which limits spectroscopic property prediction with accurate first-principles methods. Consequently, a targeted design of materials with tailored optoelectronic properties by high-throughput screening is bound to fail without efficient methods to predict molecular excited-state properties across chemical space. In this work, we present a deep neural network that predicts charged quasiparticle excitations for large and complex organic molecules with a rich elemental diversity and a size well out of reach of accurate many body perturbation theory calculations. The model exploits the fundamental underlying physics of molecular resonances as eigenvalues of a latent Hamiltonian matrix and is thus able to accurately describe multiple resonances simultaneously. The performance of this model is demonstrated for a range of organic molecules across chemical composition space and configuration space. We further showcase the model capabilities by predicting photoemission spectra at the level of the GW approximation for previously unseen conjugated molecules.


Management of Microsurgical Patients using Intraoperative Unfractionated Heparin and Thromboelastography.
Maintaining optimal coagulation is vital for successful microvascular tissue transfer. The viscoelastic thromboelastography (TEG) is a modern and dynamic method to assess a patient's coagulation status. The aim of this study was to evaluate its diagnostic capabilities of identifying microvascular complications. A retrospective chart review was conducted for the most recent 100 cases of abdominal free flap breast reconstruction of a single surgeon. Patient demographics, medical history, clinical, and operative details were documented. Thrombocyte counts, prothrombin time (PT), activated partial thromboplastin time (aPTT), and various TEG parameters were gathered for preoperative, intraoperative, and two postoperative time points. A total of hundred patients were identified, who underwent 172 abdominal-based free flaps for breast reconstruction. TEG was more dynamic compared with PT or aPTT and demonstrated borderline hypocoagulate values intraoperatively upon unfractionated heparin administration and hypercoagulate values postoperatively. In contrast, PT and aPTT demonstrated a continuously hypocoagulable state. Complications included five thrombotic events and three hematomas. The thrombotic cases had much steeper increases of TEG-G between surgery and postoperative day 2 (p = 0.049), while PT and aPTT failed to identify these patients. Of those, two resulted in flap loss (1.2%) that both occurred in patients with abdominal scars from previous surgery. The TEG is a useful adjunct for monitoring coagulation status in microsurgical breast reconstruction. When thrombosis at the anastomosis occurs, TEG correlates with a more rapid rebound from an intraoperative hypocoagulable state to a postoperative hypercoagulable state, when using the TEG. The TEG is a valuable tool for a more dynamic assessment of the patients' changing coagulation status.


Role of central ANG II receptors in stress-induced cardiovascular and hyperthermic responses in rats.
The present study was carried out using a biotelemetry system to investigate whether central angiotensin II (ANG II) is involved in stress-induced cardiovascular and body temperature responses in rats. Intracerebroventricular injections of the nonselective ANG II-receptor antagonist saralasin and of the ANG II AT1-receptor antagonist losartan attenuated both the heart rate and pressor responses to immobilization stress in a dose-dependent manner. The elevation of plasma norepinephrine and epinephrine induced by immobilization stress was also suppressed by central ANG II-receptor blockade, suggesting a general attenuation of stress-induced sympathetic nervous and adrenomedullary activity by central ANG II-receptor blockade. The hyperthermia induced by immobilization stress was attenuated by central ANG II AT1-receptor blockade in a dose-dependent manner. The effects of central saralasin on the blood pressure response induced by immobilization stress were greater in Wistar-Kyoto rats than in spontaneously hypertensive rats. The present results suggest that central ANG II AT1-receptors are involved in expression of the tachycardia and hyperthermia, as well as the pressor response, induced by immobilization stress.


Ethyl-enedi-ammonium sodium tetra-kis-[bis-(ethyl-enedi-amine-κ(2) N,N')(oxalato-κ(2) O (1),O (2))cobalt(III)] [penta-hydrogen di(phosphato-octa-deca-tungstate)] tetra-deca-hydrate.
The title compound, Na(C2H10N2)[Co(C2O4)(C2H8N2)2]4[H5(P2W18O62)2]·14H2O, prepared under hydro-thermal conditions, consists of two Dawson-type [P2W18O62](6-) anions, four isolated [Co(en)2(ox)](+) cations (en = ethyl-enedi-amine and ox = oxalate), one Na(+) cation, one [H2en](2+) cation, and a number of ordered (14) and disordered solvent water mol-ecules. The [P2W18O62](6-) polyoxidometalate anion has site symmetry 1 and contains two structurally distinct types of W atoms: viz. six W atoms on vertical pseudo-mirror planes grouped in two sets of three, and 12 equatorial W atoms that do not lie in the pseudo-mirror planes grouped in two sets of six. In each [Co(en)2(ox)](+) cation, the Co(III) ion is coordinated by four N atoms from two en ligands and two O atoms from the ox ligands, completing a distorted octa-hedral structure. The sodium cation lies on an inversion centre and additionally links the complex cations and anions. In the crystal, the various units are linked by N-H⋯O and O-H⋯O hydrogen bonds, which together with C-H⋯O hydrogen bonds form a three-dimensional structure. The contribution of a region of disordered electron density, possibly highly disordered solvent water mol-ecules, to the scattering was removed with the SQUEEZE option of PLATON [Spek (2009 ▶). Acta Cryst. D65, 148-155]. To equilibrate the charges five H(+) ions have been added to the polyoxidometalate. These H(+) ions and the disordered solvent contribution were not included in the reported mol-ecular weight and density.


Safety, tolerability, and short-term efficacy of intravenous lidocaine infusions for the treatment of chronic pain in adolescents and young adults: a preliminary report.
To provide pediatric care providers with insight into lidocaine infusions for analgesia. This retrospective review was conducted to describe lidocaine infusions for chronic refractory pain within the adolescent and young adult pain population. Although lidocaine infusions have been used for pain management in adults, their analgesic utility in the adolescent and young adult population is limited and so is the evidence for their efficacy and safety. After Institutional Board Review approval, a retrospective review of efficacy and safety data for analgesic use of lidocaine was conducted. Fifteen patients received 58 infusions with 76% receiving relief where maximum relief was seen among patients with starting pain scores ≥6/10 (mean reduction 2.3 vs. 0.5, P value = 0.006) and when a patient had three or more infusions (mean reduction 1.7 compared with 1.2). No serious side effects were encountered, but only mild or moderate side effects that did not require any intervention. Incidence of tingling or numbness and nausea or vomiting seemed to correlate with total dose of lidocaine per kilogram body weight. Patients reported reduced pain scores (6.3 ± 2.3 to 4.6 ± 2.5 before compared with after the infusion [as mean ± standard deviation]) during 80% of infusions. Our limited experience suggests that lidocaine infusions are well tolerated in the adolescent and young adult pain population, with side effects resolving quickly with interruption or discontinuation of the infusion if necessary. Future studies are warranted to examine safety, efficacy, mechanism of actions, and its long-term impact on a developing central nervous system.


Three-dimensional reconstruction of interstitial extracellular vesicles in human liver as determined by electron tomography.
Extracellular vesicles (EVs) are lipid bilayer nanoparticles involved in cell-cell communication that are released into the extracellular space by all cell types. The cargo of EVs includes proteins, lipids, nucleic acids, and metabolites reflecting their cell of origin. EVs have recently been isolated directly from solid tissues, and this may provide insights into how EVs mediate communication between cells in vivo. Even though EVs have been isolated from tissues, their point of origin when they are in the interstitial space has been uncertain. In this study, we performed three-dimensional (3D) reconstruction using transmission electron tomography of metastatic and normal liver tissues with a focus on the presence of EVs in the interstitium. After chemical fixation of the samples and subsequent embedding of tissue pieces in resin, ultrathin slices (300 nm) were cut and imaged on a 120 ekV transmission electron microscopy as a tilt series (a series of subsequent images tilted at different angles). These were then computationally illustrated in a 3D manner to reconstruct the imaged tissue volume. We identified the cells delimiting the interstitial space in both types of tissues, and small distinct spherical structures with a diameter of 30-200 nm were identified between the cells. These round structures appeared to be more abundant in metastatic tissue compared to normal tissue. We suggest that the observed spherical structures in the interstitium of the metastatic and non-metastatic liver represent EVs. This work thus provides the first 3D visualization of EVs in human tissue.


Amplicon-guided isolation and cultivation of previously uncultured microbial species from activated sludge.
Biological wastewater treatment relies on complex microbial communities that assimilate nutrients and break down pollutants in the wastewater. Knowledge about the physiology and metabolism of bacteria in wastewater treatment plants (WWTPs) may therefore be used to improve the efficacy and economy of wastewater treatment. Our current knowledge is largely based on 16S rRNA gene amplicon profiling, fluorescence in situ hybridization studies, and predictions based on metagenome-assembled genomes. Bacterial isolates are often required to validate genome-based predictions as they allow researchers to analyze a specific species without interference from other bacteria and with simple bulk measurements. Unfortunately, there are currently very few pure cultures representing the microbes commonly found in WWTPs. To address this, we introduce an isolation strategy that takes advantage of state-of-the-art microbial profiling techniques to uncover suitable growth conditions for key WWTP microbes. We furthermore demonstrate that this information can be used to isolate key organisms representing global WWTPs.


[Proteins of the "acute-phase" in prematures and newborns (author's transl)].
Three proteins of the "acute phase" (p.a.p.) were determined in the serum of 113 premature and newborns during the first 10 days of life. In all 13 cases with septicemia or severe infection high levels of c-reactive protein (CRP) and haptoglobin (Hp) were found, in 22 babies with suspected infection elevated CRP- and Hp-concentrations were found 13 and 14 times respectively. In 73 healthy newborns low levels of CRP were detectable 6 times, Hp however, was markedly elevated 25 times. Nutrition or injuries did not influence the production of Hp. The concentration of the third p.a.p. the acid alpha 1-Glycoprotein, in serum of sick and healthy neonates did not differ markedly. The indirect proof of bacterial infections by the p.a.p. is possible even in prematures under 1000 g and generally soon after the contact with pathogenes, for example in the cordblood after a previous rupture of membranes. Since CRP is the only p.a.p., that is not increased by transfusion or by age, it is by far the most suitable of the three examined p.a.p. for the detection of bacterial infections in newborn.


Porphyrias in Norway.
Porphyria is an umbrella term for a group of largely hereditary diseases that are due to defective haem synthesis. The diseases have a varied and partly overlapping range of symptoms and presentations. The commonest forms of porphyria are porphyria cutanea tarda, acute intermittent porphyria and erythropoietic protoporphyria. The purpose of this study is to provide an overview of the prevalence and pathological manifestations of porphyrias in Norway. Information on all patients registered with the Norwegian Porphyria Centre (NAPOS) up to 2012 was used to estimate the prevalence and incidence of porphyrias in Norway. Figures on symptoms, precipitating factors and follow-up routines were obtained from the Norwegian Porphyria Registry, which includes 70% of Norwegians registered with NAPOS as having porphyria. The prevalence of porphyria cutanea tarda was approximately 10 : 100,000 and that of acute intermittent porphyria approximately 4 : 100,000. The total incidence of all porphyrias was approximately 0.5-1 : 100,000 per year. Diagnostic delay, i.e. the time passing between the onset of symptoms and diagnosis, varied from 1-17 years depending on the type of porphyria. There was wide variation in the frequency with which patients with the various types of porphyria went for medical check-ups. The prevalence of acute intermittent porphyria and porphyria cutanea tarda appears to be higher in Norway than in most other countries. Data from the Norwegian Porphyria Registry makes it possible to demonstrate differences in treatment and follow-up of porphyria patients and may be used to initiate necessary measures.


Cognitive Fatigability is Independent of Subjective Cognitive Fatigue and Mood in Multiple Sclerosis.
Sustained cognitive testing is used to detect cognitive fatigability and is often considered a substitute for subjective cognitive fatigue (CF). However, the relationship between cognitive fatigability and subjective CF in people with multiple sclerosis (PwMS) remains undetermined. To explore potential associations between fatigability induced by sustained cognitive testing and subjective CF in PwMS. We gave 120 PwMS and 60 demographically matched, healthy individuals the Beck Depression Inventory-FastScreen (BDI-FS) to measure mood and the Modified Fatigue Impact Scale to measure CF. In addition, we used the Quotient ADHD Test, a sustained attention test, to measure cognitive fatigability. We also explored potential correlations between the individuals' performance on the sustained attention test and thalamic volume using recent MRI scans. Forty-one (34.2%) of the PwMS exhibited cognitive fatigability. These 41 were found to be significantly older (P=0.006), had been diagnosed with the disease for longer (P=0.03), had higher scores (P<0.001) on the Expanded Disability Status Scale, and had reduced thalamic volume (P=0.04) compared with the 79 (65.8%) PwMS not exhibiting cognitive fatigability. The PwMS exhibiting cognitive fatigability scored similarly on the BDI-FS (P=0.21) and self-reported similar rates of CF (P=0.62) as the PwMS not exhibiting cognitive fatigability. Cognitive fatigability induced by sustained cognitive testing is not an accurate clinical alternative to subjective CF. This study provides evidence to support cognitive fatigability and CF in PwMS as two distinct concepts.


Recalibrating timing behavior via expected covariance between temporal cues.
Individuals must predict future events to proactively guide their behavior. Predicting when events will occur is a critical component of these expectations. Temporal expectations are often generated based on individual cue-duration relationships. However, the durations associated with different environmental cues will often co-vary due to a common cause. We show that timing behavior may be calibrated based on this expected covariance, which we refer to as the 'common cause hypothesis'. In five experiments using rats, we found that when the duration associated with one temporal cue changes, timed-responding to other cues shift in the same direction. Furthermore, training subjects that expecting covariance is not appropriate in a given situation blocks this effect. Finally, we confirmed that this transfer is context-dependent. These results reveal a novel principle that modulates timing behavior, which we predict will apply across a variety of magnitude-expectations.


Near-infrared spectroscopy and hyperspectral imaging: non-destructive analysis of biological materials.
Near-infrared (NIR) spectroscopy has come of age and is now prominent among major analytical technologies after the NIR region was discovered in 1800, revived and developed in the early 1950s and put into practice in the 1970s. Since its first use in the cereal industry, it has become the quality control method of choice for many more applications due to the advancement in instrumentation, computing power and multivariate data analysis. NIR spectroscopy is also increasingly used during basic research performed to better understand complex biological systems, e.g. by means of studying characteristic water absorption bands. The shorter NIR wavelengths (800-2500 nm), compared to those in the mid-infrared (MIR) range (2500-15 000 nm) enable increased penetration depth and subsequent non-destructive, non-invasive, chemical-free, rapid analysis possibilities for a wide range of biological materials. A disadvantage of NIR spectroscopy is its reliance on reference methods and model development using chemometrics. NIR measurements and predictions are, however, considered more reproducible than the usually more accurate and precise reference methods. The advantages of NIR spectroscopy contribute to it now often being favoured over other spectroscopic (colourimetry and MIR) and analytical methods, using chemicals and producing chemical waste, such as gas chromatography (GC) and high performance liquid chromatography (HPLC). This tutorial review intends to provide a brief overview of the basic theoretical principles and most investigated applications of NIR spectroscopy. In addition, it considers the recent development, principles and applications of NIR hyperspectral imaging. NIR hyperspectral imaging provides NIR spectral data as a set of images, each representing a narrow wavelength range or spectral band. The advantage compared to NIR spectroscopy is that, due to the additional spatial dimension provided by this technology, the images can be analysed and visualised as chemical images providing identification as well as localisation of chemical compounds in non-homogenous samples.