TCR-independent pathways mediate the effects of antigen dose and altered peptide ligands on Th cell polarization.
We examined the role of the peptide/MHC ligand in CD4+ T cell differentiation into Th1 or Th2 cells using a TCR alphabeta transgenic mouse specific for hemoglobin (Hb)(64-76)/I-Ek. We identified two altered peptide ligands of Hb(64-76) that retain strong agonist activity but, at a given dose, induce cytokine patterns distinct from the Hb(64-76) peptide. The ability of these peptides to produce distinct cytokine patterns at identical doses is not due to an intrinsic qualitative property. Each peptide can induce Th2 cytokines at low concentrations and Th1 cytokines at high concentrations and has a unique range of concentrations at which these distinct effects occur. The pattern of cytokines produced from limiting dilution of naive T cells demonstrated that the potential to develop an individual Th1 or Th2 cell is stochastic, independent of Ag dose. We propose that the basis for the observed effects on the Th1/Th2 balance shown by the altered peptide ligands and the amount of Ag dose involves the modification of soluble factors in bulk cultures that are the driving force that polarize the population to either a Th1 or Th2 phenotype.


A machine learning approach to identify barriers in stream networks demonstrates high prevalence of unmapped riverine dams.
Restoring stream ecosystem integrity by removing unused or derelict dams has become a priority for watershed conservation globally. However, efforts to restore connectivity are constrained by the availability of accurate dam inventories which often overlook smaller unmapped riverine dams. Here we develop and test a machine learning approach to identify unmapped dams using a combination of publicly available topographic and geospatial habitat data. Specifically, we trained a random forest classification algorithm to identify unmapped dams using digitally engineered predictor variables and known dam sites for validation. We applied our algorithm to two subbasins in the Hudson River watershed, USA, and quantified connectivity impacts, as well as evaluated a range of predictor sets to examine tradeoffs between classification accuracy and model parameterization effort. The random forest classifier achieved high accuracy in predicting dam sites (true positive rate = 89%, false positive rate = 1.2%) using a subset of variables related to stream slope and presence of upstream lentic habitats. Unmapped dams were prevalent throughout the two test watersheds. In fact, existing dam inventories underestimated the true number of dams by ∼80-94%. Accounting for previously unmapped dams resulted in a 62-90% decrease in dendritic connectivity indices for migratory fishes. Unmapped dams may be pervasive and can dramatically bias stream connectivity information. However, we find that machine learning approaches can provide an accurate and scalable means of identifying unmapped dams that can guide efforts to develop accurate dam inventories, thereby informing and empowering efforts to better manage them.


Small-molecule activation at Au(iii): metallacycle construction from ethylene, water, and acetonitrile.
Incorporation of the simple, readily available, building blocks ethylene, water and acetonitrile into Au(tpy)(OCOCF3)2 (tpy = 2-(p-tolyl)pyridine) in a one-step reaction leads to high yields of a new 6-membered ring gold(iii) metallacycle complex. The metallacycle has been characterized spectroscopically and crystallographically, and the mechanism of its formation has been investigated with the aid of DFT calculations.


CT of the chest: minimal tube current required for good image quality with the least radiation dose.
We wanted to determine minimal tube current (mAs) required for consistently good image quality on conventional 10-mm collimation chest CT and effect of tube current reduction on detection of mediastinal and lung abnormalities. Tube current reduction is desirable to reduce patient radiation dose. Prospectively, 30 consecutive patients (mean weight, 68 kg; range, 34-93 kg) older than 45 undergoing conventional chest CT with standard technique (120 kVp, 400 mAs) had four additional sections imaged at reduced tube current (200, 140, 80, 20 mAs) at two levels (tracheal carina and left atrium). CT scans were evaluated in random order by two independent observers who were blinded to technical factors used. Subjective image quality was graded on a five-point scale from non-diagnostic to excellent. Visualization of mediastinal adenopathy (n = 18), pleural plaques (n = 17), effusions (n = 28), lung parenchymal nodules (n = 37), and emphysema (n = 15) were assessed. The 400 mAs scan was considered the reference standard. When compared with the reference technique (400 mAs), the first and second (200 mAs and 140 mAs) reduction levels showed no significant difference (p > .05) in subjective image quality. A significant difference (p < .001) was seen at the third and fourth (80 mAs and 20 mAs) reduction levels. However, no significant difference (p > .05) was seen in detection of mediastinal or lung parenchymal abnormalities with different tube currents. A twofold reduction in tube current (400-140 mAs) and resultant radiation dose did not cause a significant change in subjective image quality or in detection of mediastinal or lung abnormalities with conventional chest CT. One hundred forty milliampere-seconds is the minimal tube current required to provide good image quality in patients of average weight.


Effects of tension-compression nonlinearity on solute transport in charged hydrated fibrous tissues under dynamic unconfined compression.
Cartilage is a charged hydrated fibrous tissue exhibiting a high degree of tension-compression nonlinearity (i.e., tissue anisotropy). The effect of tension-compression nonlinearity on solute transport has not been investigated in cartilaginous tissue under dynamic loading conditions. In this study, a new model was developed based on the mechano-electrochemical mixture model [Yao and Gu, 2007, J. Biomech. Model Mechanobiol., 6, pp. 63-72, Lai et al., 1991, J. Biomech. Eng., 113, pp. 245-258], and conewise linear elasticity model [Soltz and Ateshian, 2000, J. Biomech. Eng., 122, pp. 576-586; Curnier et al., 1995, J. Elasticity, 37, pp. 1-38]. The solute desorption in cartilage under unconfined dynamic compression was investigated numerically using this new model. Analyses and results demonstrated that a high degree of tissue tension-compression nonlinearity could enhance the transport of large solutes considerably in the cartilage sample under dynamic unconfined compression, whereas it had little effect on the transport of small solutes (at 5% dynamic strain level). The loading-induced convection is an important mechanism for enhancing the transport of large solutes in the cartilage sample with tension-compression nonlinearity. The dynamic compression also promoted diffusion of large solutes in both tissues with and without tension-compression nonlinearity. These findings provide a new insight into the mechanisms of solute transport in hydrated, fibrous soft tissues.


Crystal polymorphism in fragment-based lead discovery of ligands of the catalytic domain of UGGT, the glycoprotein folding quality control checkpoint.
None of the current data processing pipelines for X-ray crystallography fragment-based lead discovery (FBLD) consults all the information available when deciding on the lattice and symmetry (i.e., the polymorph) of each soaked crystal. Often, X-ray crystallography FBLD pipelines either choose the polymorph based on cell volume and point-group symmetry of the X-ray diffraction data or leave polymorph attribution to manual intervention on the part of the user. Thus, when the FBLD crystals belong to more than one crystal polymorph, the discovery pipeline can be plagued by space group ambiguity, especially if the polymorphs at hand are variations of the same lattice and, therefore, difficult to tell apart from their morphology and/or their apparent crystal lattices and point groups. In the course of a fragment-based lead discovery effort aimed at finding ligands of the catalytic domain of UDP-glucose glycoprotein glucosyltransferase (UGGT), we encountered a mixture of trigonal crystals and pseudotrigonal triclinic crystals-with the two lattices closely related. In order to resolve that polymorphism ambiguity, we have written and described here a series of Unix shell scripts called CoALLA (crystal polymorph and ligand likelihood-based assignment). The CoALLA scripts are written in Unix shell and use autoPROC for data processing, CCP4-Dimple/REFMAC5 and BUSTER for refinement, and RHOFIT for ligand docking. The choice of the polymorph is effected by carrying out (in each of the known polymorphs) the tasks of diffraction data indexing, integration, scaling, and structural refinement. The most likely polymorph is then chosen as the one with the best structure refinement Rfree statistic. The CoALLA scripts further implement a likelihood-based ligand assignment strategy, starting with macromolecular refinement and automated water addition, followed by removal of the water molecules that appear to be fitting ligand density, and a final round of refinement after random perturbation of the refined macromolecular model, in order to obtain unbiased difference density maps for automated ligand placement. We illustrate the use of CoALLA to discriminate between H3 and P1 crystals used for an FBLD effort to find fragments binding to the catalytic domain of Chaetomium thermophilum UGGT.


Survivin exists in immunochemically distinct subcellular pools and is involved in spindle microtubule function.
Survivin is a member of the inhibitor of apoptosis gene family that has been implicated in both apoptosis inhibition and regulation of mitosis. However, the subcellular distribution of survivin has been controversial and variously described as a microtubule-associated protein or chromosomal passenger protein. Here, we show that antibodies directed to the survivin sequence Ala(3)-Ile(19) exclusively recognized a nuclear pool of survivin that segregated with nucleoplasmic proteins, but not with outer nuclear matrix or nuclear matrix proteins. By immunofluorescence, nuclear survivin localized to kinetochores of metaphase chromosomes, and to the central spindle midzone at anaphase. However, antibodies to Cys(57)-Trp(67) identified a cytosolic pool of survivin, which associated with interphase microtubules, centrosomes, spindle poles and mitotic spindle microtubules at metaphase and anaphase. Polyclonal antibodies recognizing survivin epitopes Ala(3)-Ile(19), Met(38)-Thr(48), Pro(47)-Phe(58) and Cys(57)-Trp(67) identified both survivin pools within the same mitotic cell. A ratio of approximately 1:6 for nuclear versus cytosolic survivin was obtained by quantitative subcellular fractionation. In synchronized cultures, cytosolic survivin abruptly increased at mitosis, physically associated with p34(cdc2), and was phosphorylated by p34(cdc2) on Thr(34), in vivo. By contrast, nuclear survivin began to accumulate in S phase, was not complexed with p34(cdc2) and was not phosphorylated on Thr(34). Intracellular loading of a polyclonal antibody to survivin caused microtubule defects and resulted in formation of multipolar mitotic spindles, but did not interfere with cytokinesis. These data demonstrate that although both reported localizations of survivin exist in mitotic cells, the preponderant survivin pool is associated with microtubules and participates in the assembly of a bipolar mitotic spindle.


p53 overexpression in human esophageal carcinoma: a correlation with tumor DNA ploidy and two parameter flow cytometric study.
Abnormalities of p53, a known tumor suppressor gene, has been detected in various human malignant neoplasms. In this study, we analyzed immunohistochemically the localization of p53 using three different antibodies, PAb1801, DO-7, and CM-1 in 49 patients with esophageal squamous cell carcinoma. We compared the expression of p53 with the clinicopathological features, proliferating cell nuclear antigen (PCNA), and DNA ploidy. In addition, we performed flow cytometric analysis of p53 using DO-7 in 33 cases in order to evaluate the correlation between p53 overexpression and DNA ploidy simultaneously. By immunohistochemistry, immunoreactive p53 was observed in the nuclei of carcinoma cells in 29 cases (59%). Relatively larger diameter tumors (more than 5cm), poorly differentiated tumors, tumors with distant lymph node metastasis, and DNA aneuploidy tumors were positive for p53 more frequently. p53-positive cases had a higher PCNA labeling index and a poorer prognosis than p53-negative cases. By two parameter flow cytometric analysis, p53 was detected in 16 cases (49%) and DNA aneuploidy was detected in 27 cases (82%). A higher p53-positive rate (15/27.56%) was observed in DNA aneuploidy cases than in DNA diploidy cases (1/6.17%). However, this difference was not statistically significant. The p53 results by immunohistochemistry and flow cytometry were the same in 25 out of 33 cases (76%). These results suggest the possible correlation between p53 abnormalities and DNA aneuploidy and that analysis of p53 protein is useful for prediction of clinical course in esophageal squamous cell carcinoma.


Exposure to radiologic contrast media and an increased risk of treated end-stage renal disease.
Radiologic contrast media can cause acute renal failure, but whether their repeated use is associated with end-stage renal disease (ESRD) is unknown. We compared 716 incident case subjects of treated ESRD with 361 control subjects, frequency matched on age, drawn from the general population (age, 20-65 years). Participants were interviewed by telephone regarding their previous exposure (before initiation of dialysis for case subjects and the study interview for control subject) to various imaging procedures. As expected, the case subjects reported having more imaging procedures of the kidneys than did control subjects. Excluding persons who had been subjected to examinations of their kidney from the analysis and adjusting for ultrasound examinations and several possible confounders, persons who had a history of one [odds ratio (OR), 2.0; 95% confidence interval (CI), 1.0, 4.0], 2 or 3 (OR, 2.6; 95% CI, 1.2,5.9), or 4 or more (OR, 3.6; 95% CI, 1.0, 12.5) radiocontrast examinations were at higher risk of treated ESRD than persons who reported not having had such procedures. Ultrasound examinations and a history of barium enema were not associated with an increased risk of treated ESRD. In the current study, a graded association was present between increasing exposure to radiologic contrast media and higher risk of treated ESRD. Whether exposure to contrast media accelerates progression to ESRD or is merely a noncausal accompaniment to multiple disease processes occurring concurrently cannot be determined from our observational data. However, if these results are confirmed in future prospective studies, they will have important clinical implications.


2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) induces peripheral blood abnormalities and plasma cell neoplasms resembling multiple myeloma in mice.
Although epidemiologic studies have suggested a possible association between occupational exposures to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and the risk of development of multiple myeloma, definitive evidence in support of this association is lacking. In the present study, we employed the Vk*Myc mouse model of multiple myeloma to assess the impact of TCDD exposure on multiple myeloma pathogenesis. TCDD induced splenomegaly and multiple peripheral blood abnormalities, including anemia and high serum IgG levels. In addition, TCDD triggered bone lytic lesions, as well as renal tubular casts, a phenomenon associated with human myeloma kidney disease. Even in wild-type C57BL/6 mice, TCDD increased serum IgG levels, induced anemia, and increased plasma cell presence in the spleen and bone marrow, hallmarks of benign monoclonal gammopathy. Lastly, TCDD induced AKT activation and the DNA damage response, key pathogenic events in myeloma pathogenesis, in animal spleen and/or bone marrow. These data indicate that TCDD accelerates monoclonal gammopathy development and promotes progression to multiple myeloma in genetically-predisposed mice. This work offers the first direct experimental evidence establishing TCDD as an environmental risk factor for monoclonal gammopathy of undetermined significance and multiple myeloma.


Can accelerometer ear tags identify behavioural changes in sheep associated with parturition?
On-animal sensor systems provide an opportunity to monitor ewes during parturition, potentially reducing ewe and lamb mortality risk. This study investigated the capacity of machine learning (ML) behaviour classification to monitor changes in sheep behaviour around the time of lambing using ear-borne accelerometers. Accelerometers were attached to 27 ewes grazing a 4.4 ha paddock. Data were then classified based on three different ethograms: (i) detection of grazing, lying, standing, walking; (ii) detection of active behaviour; and (iii) detection of body posture. Proportion of time devoted to performing each behaviour and activity was then calculated at a daily and hourly scale. Frequency of posture change was also calculated on an hourly scale. Assessment of each metric using a linear mixed-effects model was conducted for the 7 days (day scale) or 12 h (hour scale) before and after lambing. For all physical movements, regardless of the ethogram, there was a change in the days surrounding lambing. This involved either a decrease (grazing, lying, active behaviour) or peak (standing, walking) on the day of parturition, with most values returning to either pre-partum or near-pre-partum levels (all P < 0.001). Hourly changes also occurred for all behaviours (all P < 0.001), the most marked being increased walking behaviour and frequency of posture change. These findings indicate ewes were more restless around the time of parturition. Further application of this research should focus on development of algorithms that can be used to identify onset of lambing and/or time of parturition in pasture-based ewes.


Minimal Elements Required for the Formation of Respiratory Syncytial Virus Cytoplasmic Inclusion Bodies In Vivo and In Vitro.
Infection of host cells by the respiratory syncytial virus (RSV) is characterized by the formation of spherical cytoplasmic inclusion bodies (IBs). These structures, which concentrate all the proteins of the polymerase complex as well as some cellular proteins, were initially considered aggresomes formed by viral dead-end products. However, recent studies revealed that IBs are viral factories where viral RNA synthesis, i.e., replication and transcription, occurs. The analysis of IBs by electron microscopy revealed that they are membrane-less structures, and accumulated data on their structure, organization, and kinetics of formation revealed that IBs share the characteristics of cellular organelles, such as P-bodies or stress granules, suggesting that their morphogenesis depends on a liquid-liquid phase separation mechanism. It was previously shown that expression of the RSV nucleoprotein N and phosphoprotein P of the polymerase complex is sufficient to induce the formation of pseudo-IBs. Here, using a series of truncated P proteins, we identified the domains of P required for IB formation and show that the oligomeric state of N, provided it can interact with RNA, is critical for their morphogenesis. We also show that pseudo-IBs can form in vitro when recombinant N and P proteins are mixed. Finally, using fluorescence recovery after photobleaching approaches, we reveal that in cellula and in vitro IBs are liquid organelles. Our results strongly support the liquid-liquid phase separation nature of IBs and pave the way for further characterization of their dynamics.IMPORTANCE Respiratory syncytial virus (RSV) is the leading cause of lower respiratory tract illness in infants, elderly, and immunocompromised people. No vaccine or efficient antiviral treatment is available against this virus. The replication and transcription steps of the viral genome are appealing mechanisms to target for the development of new antiviral strategies. These activities take place within cytoplasmic inclusion bodies (IBs) that assemble during infection. Although expression of both the viral nucleoprotein (N) and phosphoprotein (P) allows induction of the formation of these IBs, the mechanism sustaining their assembly remains poorly characterized. Here, we identified key elements of N and P required for the scaffolding of IBs and managed for the first time to reconstitute RSV pseudo-IBs in vitro by coincubating recombinant N and P proteins. Our results provide strong evidence that the biogenesis of RSV IBs occurs through liquid-liquid phase transition mediated by N-P interactions.


[Hypoglycorrhachia in mumps meningitis (author's transl)].
Eighty cases of mumps meningitis are studied in children between the ages of one to seven years. The parotids were affected before, after and during the meningitis in all cases. In fifteen cases, when admitted to the hospital, levels of glucose were found in the C.S.F. equal or lower than 40 mg/100 ml. In three of them the hypoglycorrhachia persisted for a long time, as reported by other authors. The possible causes of low C.S.F. glucose values in meningitis are analized, not explanation for this phenomenon in viral meningitis is found.


Physical and psychosocial functioning following motor vehicle trauma: relationships with chronic pain, posttraumatic stress, and medication use.
Chronic pain and PTSD are known to hold substantial comorbidity following traumatic injury. Although pharmacological agents have been examined in the treatment of pain and PTSD individually, little is known regarding the relationship of medication use with functioning in patients with comorbid conditions. This research examined the relationships of pain, PTSD, and medication use across physical and psychosocial functioning in patients with chronic pain following motor vehicle injury (N=234). Separate analyses were conducted for opioids, SSRIs, and sedative/anxiolytics, respectively. Several relevant effects were noted: (1) Pain evidenced strong associations with reduced functioning across both physical and psychosocial domains, (2) Opioid use held interactive relationships with PTSD across both functioning domains. Specifically, opioids were associated with greater physical impairment in patients without comorbid PTSD. Opioids also were related to greater psychosocial impairment in patients without PTSD while PTSD was associated with greater impairment in patients not using opioids, (3) Opioid use evidenced a marginal interaction with pain on psychosocial functioning. Opioids were associated with greater psychosocial impairment among patients with high-pain, and high-pain was associated with greater impairment among opioid users, (4) SSRIs held a marginal interaction with PTSD such that PTSD was related to poorer psychosocial functioning only among individuals not using an SSRI, and (5) Anxiolytic use evidenced a marginal interaction with PTSD on physical functioning although no between-group differences were noted. These data suggest that PTSD symptomology may be an important consideration in determining treatment modality for patients experiencing pain subsequent to traumatic injury.


Heavy metals monitoring at a Mediterranean natural ecosystem of Central Italy. Trends in different environmental matrixes.
The study deals with the evaluation of the impact of heavy metal pollution on a Mediterranean natural ecosystem, and presents the results derived from a monitoring of heavy metals in different environmental matrixes (atmospheric dry depositions, suspended particulate matter (SPM) and stemflow of forest trees). Two sites in Castelporziano Presidential Estate (Rome), one internal and one near the sea-side, were chosen in order to assess the differences in pollutant load. Results showed that heavy metal contamination can arise from local anthropogenic activities, in particular road traffic, and long-range pollution, from industrial and artisan activities near Rome.


Laboratory examinations for the vestibular system.
In the last decades, researchers suggested that clinical assessment of labyrinthine function in detail became easy thanks to video head impulse tests (VHITs), vestibular evoked myogenic potential test (VEMP) and video-oculography (VOG). It has been argued that they can replace electronystagmography, the caloric and rotatory chair tests. This review addresses the latest evaluations of these tests and the opportunities they offer, but also the limitations in clinical practice. The VHIT and suppression head impulse test (SHIMP) are under ideal circumstances able to accurately identify deficits of the VOR in 3D. However, in a relevant part of the patient population, pupil tracking is inaccurate, video-goggles slip and VOR quantification is problematic. The dissociation between the VHIT and caloric test suggests that these tests are complementary. A new 3D-VOG technique claims to quantify eye torsion better than before, opening multiple diagnostic possibilities. VEMPs remain difficult to standardize. Variability in normal cervical vestibular-evoked myogenic potential amplitude is large. VEMPs become smaller or absent with age, raising questions of whether there is a lower normal limit at all. Recent research shows that the labyrinth is directly stimulated in the MRI offering new opportunities for diagnostics and research. In clinical practice, the VHIT, SHIMP, VEMP and new 3D-VOG techniques improve diagnostic power. Unfortunately, technical issues or variability prevent reliable quantitative evaluation in a part of the regular patient population. The traditional caloric and rotatory chair test can still be considered as valuable complementary tests.


Cortical and hippocampal correlates of deliberation during model-based decisions for rewards in humans.
How do we use our memories of the past to guide decisions we've never had to make before? Although extensive work describes how the brain learns to repeat rewarded actions, decisions can also be influenced by associations between stimuli or events not directly involving reward - such as when planning routes using a cognitive map or chess moves using predicted countermoves - and these sorts of associations are critical when deciding among novel options. This process is known as model-based decision making. While the learning of environmental relations that might support model-based decisions is well studied, and separately this sort of information has been inferred to impact decisions, there is little evidence concerning the full cycle by which such associations are acquired and drive choices. Of particular interest is whether decisions are directly supported by the same mnemonic systems characterized for relational learning more generally, or instead rely on other, specialized representations. Here, building on our previous work, which isolated dual representations underlying sequential predictive learning, we directly demonstrate that one such representation, encoded by the hippocampal memory system and adjacent cortical structures, supports goal-directed decisions. Using interleaved learning and decision tasks, we monitor predictive learning directly and also trace its influence on decisions for reward. We quantitatively compare the learning processes underlying multiple behavioral and fMRI observables using computational model fits. Across both tasks, a quantitatively consistent learning process explains reaction times, choices, and both expectation- and surprise-related neural activity. The same hippocampal and ventral stream regions engaged in anticipating stimuli during learning are also engaged in proportion to the difficulty of decisions. These results support a role for predictive associations learned by the hippocampal memory system to be recalled during choice formation.


Motor asymmetry in goats during a stepping task.
Behavioural lateralization consists of perceptual and motor lateralization and provides adaptive advantages such as a general increase in brain efficiency. Motor laterality refers to the preferred use of either left or right limbs or organs to perform a specific task. We investigated motor laterality in goats (Capra hircus), using the First-stepping Task. During this task, the first foreleg used to step off a board after standing with both forelimbs was recorded. Subjects varied individually in their expression of motor lateralization with 36.6% of subjects showing individual-level asymmetries. However, goats as a group did not show a preference for a specific foreleg or lateralization in general. Our results support the hypothesis that the need to coordinate behaviour among conspecifics might be important for determining the presence of lateralization at the population level. We suggest that future research investigates how social complexity might affect population-level asymmetries, and whether stimuli with high emotional valence impact on lateralization presence and level (i.e., individual or population).


Delayed induction of type I and III interferons mediates nasal epithelial cell permissiveness to SARS-CoV-2.
The nasal epithelium is a plausible entry point for SARS-CoV-2, a site of pathogenesis and transmission, and may initiate the host response to SARS-CoV-2. Antiviral interferon (IFN) responses are critical to outcome of SARS-CoV-2. Yet little is known about the interaction between SARS-CoV-2 and innate immunity in this tissue. Here we apply single-cell RNA sequencing and proteomics to a primary cell model of human nasal epithelium differentiated at air-liquid interface. SARS-CoV-2 demonstrates widespread tropism for nasal epithelial cell types. The host response is dominated by type I and III IFNs and interferon-stimulated gene products. This response is notably delayed in onset relative to viral gene expression and compared to other respiratory viruses. Nevertheless, once established, the paracrine IFN response begins to impact on SARS-CoV-2 replication. When provided prior to infection, recombinant IFNβ or IFNλ1 induces an efficient antiviral state that potently restricts SARS-CoV-2 viral replication, preserving epithelial barrier integrity. These data imply that the IFN-I/III response to SARS-CoV-2 initiates in the nasal airway and suggest nasal delivery of recombinant IFNs to be a potential chemoprophylactic strategy.


Activation of B lymphocyte maturation by a human follicular dendritic cell line, FDC-1.
Previously, we described the characteristics of a cell line that is derived from a low density fraction of human tonsillar cells and, on the basis of a number of criteria, is related to follicular dendritic cells (FDC). This line, FDC-1, binds B lymphocytes and not T lymphocytes, and promotes anti-Ig- or anti-CD40-induced B cell proliferation. In this work, we show that culturing B cells with small numbers of FDC-1 cells leads to significant production of IL-6 and of both IgM and IgG. As few as 50 to 100 FDC-1 augmented B cell IgM production by 10- to 100-fold. Although fixed FDC-1 cells, unlike live FDC-1 cells, do not stimulate Ig production, cell contact is not required for all FDC-dependent Ig production. Supernatants from cultured FDC-1 cells can also stimulate B cells to produce IgM, suggesting that FDC produce a soluble B cell stimulating factor(s). Augmentation of FDC-dependent IgM production by either IL-6 or IL-7 and augmentation of FDC-dependent IgG production by IL-4 does require FDC-1 cells to be in contact with B cells. When the effects of FDC-1 cells were compared with those of epithelial cell lines and human foreskin fibroblasts (HFF), both FDC-1 cells and HFF induced B cells to produce IgM. FDC-1, unlike HFF, were positive for CD40, CD54, CD73, CD74, and nerve growth factor receptor (NGFR), and unlike HFF, but like certain stromal cells, FDC-1 cells also expressed smooth muscle actin, and a novel marker for stromal cells, BST-1. The possible relationship of FDC-1 cells and FDC in general to a fibroblast/stromal cell lineage is discussed.


The Effects of Altitude Training on Erythropoietic Response and Hematological Variables in Adult Athletes: A Narrative Review.
Background: One of the goals of altitude training is to increase blood oxygen-carrying capacity in order to improve sea-level endurance performance in athletes. The elevated erythropoietin (EPO) production in hypoxia is a key factor in the achievement of enhanced hematological variables. The level of the EPO increase and acceleration of erythropoiesis depend on the duration of exposure and degree of hypoxia. Furthermore, many other factors may affect the hematological response to altitude training. Aim: The purpose of this narrative review was to: (1) analyze the kinetics of EPO and hematological variables during and after altitude training; (2) summarize the current state of knowledge about the possible causes of individual or cohort differences in EPO and hematological response to altitude training; (3) formulate practical guidelines for athletes to improve the efficiency of altitude training. Methods: A narrative review was performed following an electronic search of the databases PubMed/MEDLINE and SPORTDiscus via EBSCO for all English-language articles published between 1997 and 2017. Results: Complete unification of results from studies on EPO kinetics was difficult due to different time and frequency of blood sampling by different researchers during and after altitude training, but the data presented in the reviewed literature allowed us to detect certain trends. The results of the reviewed studies were divergent and indicated either increase or no change of hematological variables following altitude training. Factors that may affect the hematological response to altitude training include hypoxic dose, training content, training background of athletes, and/or individual variability of EPO production. Conclusions: Despite the potential benefits arising from altitude training, its effectiveness in improving hematological variables is still debatable. Further research and better understanding of factors influencing the response to altitude, as well as factors affecting the suitable measurement and interpretation of study results, are needed.


Dissemination of the CTX-M-25 family beta-lactamases among Klebsiella pneumoniae, Escherichia coli and Enterobacter cloacae and identification of the novel enzyme CTX-M-41 in Proteus mirabilis in Israel.
The CTX-M-25 family of beta-lactamases is a closely related family of enzymes found rarely in the world. We aimed to describe the occurrence and to understand the dissemination of this extended-spectrum beta-lactamase family among Enterobacteriaceae strains in our hospital. Fifty-four CTX-M-producing Enterobacteriaceae strains collected from 2000 to 2005 were screened for bla(CTX-M-25) genes by PCR and sequencing. Genetic relatedness was analysed by PFGE. Antibiotic susceptibilities were determined by VITEK-2. Plasmids encoding bla(CTX-M-25)-type genes were isolated, transformed and analysed by Southern blot using a bla(CTX-M-25) probe. Chromosomal location of bla(CTX-M-25)-type was studied by I-CeuI restriction analysis. The bla(CTX-M-25) genetic environment was characterized by PCR mapping and partial sequencing. Ten out of 54 CTX-M-producing isolates (18.5%) carried bla(CTX-M-25) genes, including Klebsiella pneumoniae (n = 4), Escherichia coli (n = 3), Enterobacter cloacae (n = 1) and Proteus mirabilis (n = 2). Isolates were genetically unrelated. Four beta-lactamases were found: CTX-M-25, CTX-M-26, CTX-M-39 and CTX-M-41, a new member of the family (accession no. DQ023162) that differed from CTX-M-25 in three amino acids, Ala80Val, Val106Ile and Ile126Ser. bla(CTX-M-25)-type genes were plasmid-mediated in all genera but P. mirabilis, organized in a class I integron and located downstream of an ISEcp1 element. The genes were encoded on different plasmids with varying degree of similarities. Several antibiotic-resistant determinants conferring resistance to trimethoprim and aminoglycosides existed on the same integron. bla(CTX-M-25) exists in Israel in different enteric species. Spread of these enzymes within and between species is due to transfer of plasmids with common regions and by dissemination of determinants encoding these genes. CTX-M-41, a novel member of this family, was identified in the chromosome of P. mirabilis.


Differential effect of clomipramine on habituation and prepulse inhibition in dominant versus subordinate rats.
Many patients with depression have comorbidities associated with an impairment of sensorimotor gating, such as e.g. schizophrenia, Parkinson Disease, or Alzheimer disease. Anti-depressants like clomipramine that modulate serotonergic or norepinephrinergic neurotransmission have been shown to impact sensorimotor gating, it is therefore important to study potential effects of clomipramine in order to rule out an exacerbation of sensorimotor gating impairment. Prior studies in animals and humans have been inconclusive. Since serotonin and norepinephrine levels are closely related to anxiety and stress levels and therefore to the social status of an animal, we tested the hypothesis that acute and chronic effects of clomipramine on sensorimotor gating are different in dominant versus subordinate rats, which might be responsible for conflicting results in past animal studies. We used habituation and prepulse inhibition (PPI) of the acoustic startle response as operational measures of sensorimotor gating. After establishing the dominant animal in pair-housed male rats, we injected clomipramine for two weeks and measured acute effects on baseline startle, habituation and PPI after the first injection and chronic effects at the end of the two weeks. Chronic treatment with clomipramine significantly increased habituation in subordinate rats, but had no effect on habituation in dominant animals. Furthermore, PPI was slightly enhanced in subordinate rats upon chronic treatment while no changes occurred in dominant animals. We conclude that the social status of an animal, and therefore the basic anxiety/stress level determines whether or not clomipramine has a beneficial effect on sensorimotor gating and discuss possible underlying mechanisms.


Restriction fingerprinting and serology in a small outbreak of B15 meningococcal disease among Norwegian soldiers.
In September 1981 a soldier died from meningococcal septicemia in a military camp in Mid-Norway. Soon afterwards one of his room-mates was transferred to a military camp in Northern-Norway where he shared sleeping quarters (room 7D) with 5 other soldiers of whom 2 fell ill with meningococcal disease 1 month later. Throat cultures were obtained from all 128 soldiers at the military camp in Northern-Norway; 41 (32%) harboured meningococci in their throats. The 3 invasive isolates and the isolates from the 4 healthy carriers at room 7D were all group B and type 15 meningococci. However, by DNA fingerprinting we could identify at least 2, probably 3, different individual strains among these 7 isolates. None of these strains were isolated from soldiers outside room 7D. By use of a B15 whole-bacterium ELISA method we showed that the levels of antimeningococcal IgG antibodies in the sera of the two cases at room 7D were low (18 and 28 OD units) compared with the mean IgG levels in the sera of their 4 healthy room mates (1150 OD units) and the mean IgG in the sera from all healthy soldiers (472 OD units).


Use of a multiplex polymerase chain reaction assay in the antemortem diagnosis of toxoplasmosis and neosporosis in the central nervous system of cats and dogs.
To develop a multiplex polymerase chain reaction (PCR) assay for the detection of Toxoplasma gondii and Neospora caninum DNA in canine and feline biological samples. SAMPLE POPULATION; Biological samples from 7 cats with systemic (n = 4) or CNS (3) toxoplasmosis, 6 dogs with neospora- or toxoplasma-associated encephalitis, and 11 animals with nonprotozoal disease. Primers for T gondii, N caninum, and the canine ferritin gene (dogs) or feline histone 3.3 gene (cats) were combined in a single PCR assay. The DNA was extracted from paraffin-embedded brain tissue, CSF, or skeletal muscle. The PCR products with positive results were cloned, and sequence identity was confirmed. Of 7 cats and 4 dogs with immunohistochemical or serologic evidence of toxoplasmosis, PCR results were positive for all cats and 3 dogs for T gondii, and positive for T gondii and N caninum for 1 dog. Another dog had negative PCR results for both parasites. Of 2 dogs with immunohistochemical or serologic evidence of neosporosis, PCR results were positive for 1 for N caninum and positive for the other for T gondii. All negative-control samples yielded negative results for T gondii and N caninum on the PCR assay. Standard tests for toxoplasmosis or neosporosis associated with the CNS rely on serologic, histologic, or immunohistochemical analysis and can be difficult to interpret. The multiplex PCR assay with built-in control reactions could be a complementary clinical tool for the antemortem diagnosis of toxoplasmosis or neosporosis associated with the CNS.


Weak intermolecular interactions in gas-phase nuclear magnetic resonance.
Gas-phase nuclear magnetic resonance (NMR) spectra demonstrating the effect of weak intermolecular forces on the NMR shielding constants of the interacting species are reported. We analyse the interaction of the molecular hydrogen isotopomers with He, Ne, and Ar, and the interaction in the He-CO(2) dimer. The same effects are studied for all these systems in the ab initio calculations. The comparison of the experimental and computed shielding constants is shown to depend strongly on the treatment of the bulk susceptibility effects, which determine in practice the pressure dependence of the experimental values. Best agreement of the results is obtained when the bulk susceptibility correction in rare gas solvents is evaluated from the analysis of the He-rare gas interactions, and when the shielding of deuterium in D(2)-rare gas systems is considered.


The 3-hour therapy criterion: a challenge for rehabilitation facilities.
Rehabilitation hospitals and units must meet certain requirements to be reimbursed by Medicare. Section 211 of the Medicare Hospital Manual specifies that at least 3 hours a day of physical and/or occupational therapy must be provided to each patient in addition to other required therapies and services. This article discusses the political and professional context surrounding the implementation of these guidelines and some of the practical issues that managers must address in the process. A case report is used to illustrate the management strategies and specific methods that have been implemented in the Occupational Therapy Department at the Rehabilitation Institute of Chicago in response to the 3-hour criterion. Data are presented to indicate how these efforts have contributed to raising our level of compliance with the criterion during the 1985-1986 fiscal year. The compliance data are discussed in relation to variation in both the number of patients and staff productivity over the course of the year.


Biphasic alterations in coronary smooth muscle Ca(2+) regulation in a repeat cross-sectional study of coronary artery disease severity in metabolic syndrome.
Coronary artery disease (CAD) is progressive, classified by stages of severity. Alterations in Ca(2+) regulation within coronary smooth muscle (CSM) cells in metabolic syndrome (MetS) have been observed, but there is a lack of data in relatively early (mild) and late (severe) stages of CAD. The current study examined alterations in CSM Ca(2+) regulation at several time points during CAD progression. MetS was induced by feeding an excess calorie atherogenic diet for 6, 9, or 12 months and compared to age-matched lean controls. CAD was measured with intravascular ultrasound (IVUS). Intracellular Ca(2+) was assessed with fura-2. IVUS revealed that the extent of atherosclerotic CAD correlated with the duration on atherogenic diet. Fura-2 imaging of intracellular Ca(2+) in CSM cells revealed heightened Ca(2+) signaling at 9 months on diet, compared to 6 and 12 months, and to age-matched lean controls. Isolated coronary artery rings from swine fed for 9 months followed the same pattern, developing greater tension to depolarization, compared to 6 and 12 months (6 months = 1.8 ± 0.6 g, 9 months = 5.0 ± 1.0 g, 12 months = 0.7 ± 0.1 g). CSM in severe atherosclerotic plaques showed dampened Ca(2+) regulation and decreased proliferation compared to CSM from the wall. These CSM Ca(2+) regulation data from several time points in CAD progression and severity help to resolve the controversy regarding up-vs. down-regulation of CSM Ca(2+) regulation in previous reports. These data are consistent with the hypothesis that alterations in sarcoplasmic reticulum Ca(2+) contribute to progression of atherosclerotic CAD in MetS.


Aberrant recruitment of the nuclear receptor corepressor-histone deacetylase complex by the acute myeloid leukemia fusion partner ETO.
Nuclear receptor corepressor (CoR)-histone deacetylase (HDAC) complex recruitment is indispensable for the biological activities of the retinoic acid receptor fusion proteins of acute promyelocytic leukemias. We report here that ETO (eight-twenty-one or MTG8), which is fused to the acute myelogenous leukemia 1 (AML1) transcription factor in t(8;21) AML, interacts via its zinc finger region with a conserved domain of the corepressors N-CoR and SMRT and recruits HDAC in vivo. The fusion protein AML1-ETO retains the ability of ETO to form stable complexes with N-CoR/SMRT and HDAC. Deletion of the ETO C terminus abolishes CoR binding and HDAC recruitment and severely impairs the ability of AML1-ETO to inhibit differentiation of hematopoietic precursors. These data indicate that formation of a stable complex with CoR-HDAC is crucial to the activation of the leukemogenic potential of AML1 by ETO and suggest that aberrant recruitment of corepressor complexes is a general mechanism of leukemogenesis.


What do we know about the impact of sexual violence on health and health behaviour of women in Estonia?
Sexual violence against women is a major public health issue and a breach of human rights. Although various consequences of sexual violence on health have been described in a large number of scientific publications, very little is known about this topic in Estonia. The aim of this study was to examine the prevalence of sexual violence and associations between exposure to sexual violence and risky health and sexual behaviours among women in Estonia. A population-based cross-sectional study was carried out in Estonia in 2014. Self-reported data regarding selected indicators of risky health and sexual behaviours were collected from 1670 women, aged 18-44 years, via a self-administered questionnaire. To measure the prevalence of sexual violence, questions from the NorVold Abuse Questionnaire were included. Chi-square and multivariate logistic regression were used to analyse the data. Of the respondents, 22.7% (n = 379) reported being exposed to sexual violence during their lifetime, and over half of these women had had these experiences before the age of 18. Statistically significant associations were found between sexual violence and smoking (adjusted odds ratio (AOR) 1.32, 95% CI 1.03-1.70), alcohol consumption (AOR 1.52, 95% CI 1.18-1.95), illicit drug use (AOR 2.21, 95% CI 1.70-2.89), sexual intercourse for money or other material reward (AOR 3.51, 95% CI 1.62-7.61), concurrent sexual relationships (AOR 2.64; 95% CI 1.80-3.86), and being diagnosed with sexually transmitted infections (AOR 1.48, 95% CI 1.09-2.01). In Estonia, sexual violence against women is widespread and is associated with several risky health and sexual behaviours. Efforts should be made, both among the general public and professionals, to raise awareness regarding the prevalence and negative impact of sexual violence. Women who have been exposed to sexual violence are in need of professional medical, legal and psychological help free from prejudice to help them recover from such traumatic events.


White-light generation using spatially-structured beams of femtosecond radiation.
We studied white-light generation in water using spatially- structured beams of femtosecond radiation. By changing the transverse spatial phase of an initial Gaussian beam with a 1D spatial light modulator to that of an Hermite-Gaussian (HGn,m) mode, we were able to generate beams exhibiting phase discontinuities and steeper intensity gradients. When the spatial phase of an initial Gaussian beam (showing no significant white-light generation) was changed to that of a HG01, or HG11 mode, significant amounts of white-light were produced. Because self-focusing is known to play an important role in white-light generation, the self-focusing lengths of the resulting transverse intensity profiles were used to qualitatively explain this production. Distributions of the laser intensity for beams having step-wise spatial phase variations were modeled using the Fresnel-Kirchhoff integral in the Fresnel approximation and found to be in good agreement with experiment.


[Methodological problems in the neurophysiology of behavior].
The methodological problems of correlation of analytical and synthetical approaches (reduction and integration) in research on the brain mechanisms of behaviour are considered. The correlation of plasticity of polyfunctional brain structures with relative stability of behavioral defects after the damage of the same structures are discussed. The teaching of A. A. Ukhtomskiĭ on the dominant as a complex of brain mechanisms securing the active behavioral nature may be a methodological basis for purposeful experimental studies.


Characterization of the early steps of human parvovirus B19 infection.
The early steps of human parvovirus B19 (B19V) infection were investigated in UT7/Epo cells. B19V and its receptor globoside (Gb4Cer) associate with lipid rafts, predominantly of the noncaveolar type. Pharmacological disruption of the lipid rafts inhibited infection when the drug was added prior to virus attachment but not after virus uptake. B19V is internalized by clathrin-dependent endocytosis and spreads rapidly throughout the endocytic pathway, reaching the lysosomal compartment within minutes, where a substantial proportion is degraded. B19V did not permeabilize the endocytic vesicles, indicating a mechanism of endosomal escape without apparent membrane damage. Bafilomycin A(1) (BafA1) and NH(4)Cl, which raise endosomal pH, blocked the infection by preventing endosomal escape, resulting in a massive accumulation of capsids in the lysosomes. In contrast, in the presence of chloroquine (CQ), the transfer of incoming viruses from late endosomes to lysosomes was prevented; the viral DNA was not degraded; and the infection was boosted. In contrast to the findings for untreated or BafA1-treated cells, the viral DNA was progressively associated with the nucleus in CQ-treated cells, reaching a plateau by 3 h postinternalization, a time coinciding with the initiation of viral transcription. At this time, more than half of the total intracellular viral DNA was associated with the nucleus; however, the capsids remained extranuclear. Our studies provide the first insight into the early steps of B19V infection and reveal mechanisms involved in virus uptake, endocytic trafficking, and nuclear penetration.


Influenza B and C virus NEP (NS2) proteins possess nuclear export activities.
Nucleocytoplasmic transport of viral ribonucleoproteins (vRNPs) is an essential aspect of the replication cycle for influenza A, B, and C viruses. These viruses replicate and transcribe their genomes in the nuclei of infected cells. During the late stages of infection, vRNPs must be exported from the nucleus to the cytoplasm prior to transport to viral assembly sites on the cellular plasma membrane. Previously, we demonstrated that the influenza A virus nuclear export protein (NEP, formerly referred to as the NS2 protein) mediates the export of vRNPs. In this report, we suggest that for influenza B and C viruses the nuclear export function is also performed by the orthologous NEP proteins (formerly referred to as the NS2 protein). The influenza virus B and C NEP proteins interact in the yeast two-hybrid assay with a subset of nucleoporins and with the Crm1 nuclear export factor and can functionally replace the effector domain from the human immunodeficiency virus type 1 Rev protein. We established a plasmid transfection system for the generation of virus-like particles (VLPs) in which a functional viral RNA-like chloramphenicol acetyltransferase (CAT) gene is delivered to a new cell. VLPs generated in the absence of the influenza B virus NEP protein were unable to transfer the viral RNA-like CAT gene to a new cell. From these data, we suggest that the nuclear export of the influenza B and C vRNPs are mediated through interaction between NEP proteins and the cellular nucleocytoplasmic export machinery.


Surgical Practice in the Shadow of COVID-19 Outbreak.
COVID-19 epidemic rapidly spread all around the world with over 1500 thousand infected cases and 95000 deaths. This rapid pandemic may overwhelm health care capacity and shortage of resources is a major concern. Literature provided guidelines on management of COVID-19 patients but healthcare service to the normal population should be continued meanwhile. Health system should act immediately and wisely to support essential surgical care while fighting against COVID-19. We conducted a comprehensive search in the major data bases since 2020, using the combination of MeSH words of "COVID-19 " and "surgery" and finally 34 full texts entered to data extraction phase to define a plan for surgical practice during COVID-19 pandemic. Healthcare workers are at the higher risk of contamination by COVID-19 especially in early stage of outbreak when they were not aware of the different aspects of COVID-19 pandemic. All healthcare staff must be trained to properly use PPE. All patients have to be screened at the hospital triage. All elective surgical interventions must be postponed. Operation room is considered as a place with high risk of cross infection so the highest level of protection should be maintained. Anesthesia, endoscopy and oral surgery are considered as aerosol producing procedures with very high risk of contamination. There is not any evidence to support the risk of infection trough blood products. Postoperative respiratory problems are more common among COVID-19 patients that may increases the estimated risk of morbidity and mortality. COVID-19 pandemic is a dynamic challenge for health system to save the healthcare staff and equipment resources by timely decisions. Healthcare workers are at the higher risk of contamination by COVID-19 especially in early phase of epidemic when the protection is sub-optimal.


Comparative analysis of membrane-associated fusion peptide secondary structure and lipid mixing function of HIV gp41 constructs that model the early pre-hairpin intermediate and final hairpin conformations.
Fusion between viral and host cell membranes is the initial step of human immunodeficiency virus infection and is mediated by the gp41 protein, which is embedded in the viral membrane. The approximately 20-residue N-terminal fusion peptide (FP) region of gp41 binds to the host cell membrane and plays a critical role in fusion catalysis. Key gp41 fusion conformations include an early pre-hairpin intermediate (PHI) characterized by extended coiled-coil structure in the region C-terminal of the FP and a final hairpin state with compact six-helix bundle structure. The large "N70" (gp41 1-70) and "FP-Hairpin" constructs of the present study contained the FP and respectively modeled the PHI and hairpin conformations. Comparison was also made to the shorter "FP34" (gp41 1-34) fragment. Studies were done in membranes with physiologically relevant cholesterol content and in membranes without cholesterol. In either membrane type, there were large differences in fusion function among the constructs with little fusion induced by FP-Hairpin, moderate fusion for FP34, and very rapid fusion for N70. Overall, our findings support acceleration of gp41-induced membrane fusion by early PHI conformation and fusion arrest after folding to the final six-helix bundle structure. FP secondary structure at Leu7 of the membrane-associated constructs was probed by solid-state nuclear magnetic resonance and showed populations of molecules with either beta-sheet or helical structure with greater beta-sheet population observed for FP34 than for N70 or FP-Hairpin. The large differences in fusion function among the constructs were not obviously correlated with FP secondary structure. Observation of cholesterol-dependent FP structure for fusogenic FP34 and N70 and cholesterol-independent structure for non-fusogenic FP-Hairpin was consistent with membrane insertion of the FP for FP34 and N70 and with lack of insertion for FP-Hairpin. Membrane insertion of the FP may therefore be associated with the early PHI conformation and FP withdrawal with the final hairpin conformation.


Enabling Spike-Based Backpropagation for Training Deep Neural Network Architectures.
Spiking Neural Networks (SNNs) have recently emerged as a prominent neural computing paradigm. However, the typical shallow SNN architectures have limited capacity for expressing complex representations while training deep SNNs using input spikes has not been successful so far. Diverse methods have been proposed to get around this issue such as converting off-the-shelf trained deep Artificial Neural Networks (ANNs) to SNNs. However, the ANN-SNN conversion scheme fails to capture the temporal dynamics of a spiking system. On the other hand, it is still a difficult problem to directly train deep SNNs using input spike events due to the discontinuous, non-differentiable nature of the spike generation function. To overcome this problem, we propose an approximate derivative method that accounts for the leaky behavior of LIF neurons. This method enables training deep convolutional SNNs directly (with input spike events) using spike-based backpropagation. Our experiments show the effectiveness of the proposed spike-based learning on deep networks (VGG and Residual architectures) by achieving the best classification accuracies in MNIST, SVHN, and CIFAR-10 datasets compared to other SNNs trained with a spike-based learning. Moreover, we analyze sparse event-based computations to demonstrate the efficacy of the proposed SNN training method for inference operation in the spiking domain.


Individual and neighborhood risk factors of hospital admission and death during the COVID-19 pandemic: a population-based cohort study.
The coronavirus disease 2019 (COVID-19) disproportionately affects minority populations in the USA. Sweden - like other Nordic countries - have less income and wealth inequality but lacks data on the socioeconomic impact on the risk of adverse outcomes due to COVID-19. This population-wide study from March 2020 to March 2022 included all adults in Stockholm, except those in nursing homes or receiving in-home care. Data sources include hospitals, primary care (individual diagnoses), the Swedish National Tax Agency (death dates), the Total Population Register "RTB" (sex, age, birth country), the Household Register (size of household), the Integrated Database For Labor Market Research "LISA" (educational level, income, and occupation), and SmiNet (COVID data). Individual exposures include education, income, type of work and ability to work from home, living area and living conditions as well as the individual country of origin and co-morbidities. Additionally, we have data on the risks associated with living areas. We used a Cox proportional hazards model and logistic regression to estimate associations. Area-level covariates were used in a principal component analysis to generate a measurement of neighborhood deprivation. As outcomes, we used hospitalization and death due to COVID-19. Among the 1,782,125 persons, male sex, comorbidities, higher age, and not being born in Sweden increase the risk of hospitalization and death. So does lower education and lower income, the lowest incomes doubled the risk of death from COVID-19. Area estimates, where the model includes individual risks, show that high population density and a high percentage of foreign-born inhabitants increased the risk of hospitalization. Segregation and deprivation are public health issues elucidated by COVID-19. Neighborhood deprivation, prevalent in Stockholm, adds to individual risks and is associated with hospitalization and death. This finding is paramount for governments, agencies, and healthcare institutions interested in targeted interventions.


Biophysical characterization of higher plant Rubisco activase.
Rubisco activase (Rca) is a chaperone-like protein of the AAA+ family, which uses mechano-chemical energy derived from ATP hydrolysis to release tightly bound inhibitors from the active site of the primary carbon fixing enzyme ribulose 1,5-bisphosphate oxygenase/carboxylase (Rubisco). Mechanistic and structural investigations of Rca have been hampered by its exceptional thermolability, high degree of size polydispersity and propensity towards subunit aggregation. In this work, we have characterized the thermal stability and self-association behavior of recombinant Rca preparations, and have developed ligand screening methods. Thermal denaturation profiles generated by circular dichroism indicate that creosote and tobacco short-form Rcas are the most stable proteins examined, with an estimated mid-point temperature of 45-47°C for protein denaturation. We demonstrate that ADP provides a higher degree of stabilization than ATP, that magnesium ions have a small stabilizing effect on ATP-bound, but a significant destabilizing effect on ADP-bound Rca, and that phosphate provides weak stabilization of the ADP-bound form of the protein. A dimeric species was identified by size-exclusion chromatography, suggesting that the two-subunit module may comprise the basic building block for larger assemblies. Evidence is provided that chromatographic procedures reflect non-equilibrium multimeric states. Dynamic light scattering experiments performed on nucleotide-bearing Rca support the notion that several larger, highly polydisperse assembly states coexist over a broad concentration range. No significant changes in aggregation are observed upon replacement of ADP with ATP. However, in the absence of nucleotides, the major protein population appears to consist of a monodisperse oligomer smaller than a hexamer.


Nasal polyps, bronchial asthma and aspirin sensitivity.
The ASA triad comprises bronchial asthma, acetylsalicylic acid (ASA) sensitivity and nasal polyps. It presents as chronic rhinitis followed by bronchial asthma and ASA sensitivity, and later nasal polyps. The pathogenesis of the ASA triad may involve interrelationships between disease in the upper and lower airway and hypersensitivity to cyclo-oxygenase inhibiting medications. Treatment of the nasal polyps has been shown to improve the patients' asthma.


HIV Care Initiation Delay Among Rural Residents in the Southeastern United States, 1996 to 2012.
Delaying HIV care initiation may lead to greater morbidity, mortality, and further HIV transmission. Rural residence may be associated with delayed diagnosis and linkage to care, with negative clinical outcomes. To examine the association between rural patient residence and CD4 cell count at HIV care initiation in a large HIV clinical cohort in the Southeastern United States. We included HIV-infected patients who initiated care between 1996 and 2012 with a geocodable address and no previous history of HIV clinical care. Patient residence was categorized as urban or rural using United States Department of Agriculture Rural Urban Commuting Area codes. Multivariable linear regression models were fit to estimate the association between patient residence and CD4 cell count at HIV care initiation. Among 1396 patients who met study inclusion criteria, 988 had a geocodable address. Overall, 35% of patients resided in rural areas and presented to HIV care with a mean CD4 cell count of 351 cells/mm (SD, 290). Care initiation mean CD4 cell counts increased from 329 cells/mm (SD, 283) in 1996-2003 to 391 cells/mm (SD, 292) in 2008-2012 (P = 0.006). Rural in comparison with urban patients presented with lower CD4 cell counts with an unadjusted and adjusted mean difference of -48 cells/mm [95% confidence interval, -86 to -10) and -37 cells/mm (95% confidence interval: -73 to -2), respectively, consistently observed across calendar years. HIV care initiation at low CD4 cell counts was common in this Southeastern US cohort and more common among rural area residents.


Eps15R is required for bone morphogenetic protein signalling and differentially compartmentalizes with Smad proteins.
Transforming growth factor β superfamily members signal through Smad transcription factors. Bone morphogenetic proteins (BMPs) act via Smads 1, 5 and 8 and TGF-βs signal through Smads 2 and 3. The endocytic adaptor protein Eps15R, or 'epidermal growth factor (EGF) receptor pathway substrate 15-related protein' is a component of EGF signal transduction, mediating internalization of the EGF receptor. We show that it interacts with Smad proteins, is required for BMP signalling in animal caps and stimulates Smad1 transcriptional activity. This function resides in the Asp-Pro-Phe motif-enriched 'DPF domain' of Eps15R, which activates transcription and antagonizes Smad2 signalling. In living cells, Eps15R segregates into spatially distinct regions with different Smads, indicating an unrecognized level of Smad compartmentalization.


Cognitive function in 1736 participants in NINDS Exploratory Trials in PD Long-term Study-1.
Clinical cohort studies suggest that mild cognitive impairment (MCI) is common in early Parkinson's disease (PD). The objectives of this paper were to describe cognitive function in a large clinical trial of early treated PD patients at baseline and over time using two brief cognitive screening tests. In total 1741 participants were enrolled in the NINDS Exploratory Trials in Parkinson's disease (NET-PD) Long-term Study-1 (LS-1). The Symbol Digit Modalities Test (SDMT) was collected annually. The SCales for Outcomes in PArkinson's disease-COGnition (SCOPA-COG) was collected at baseline and at year 5. The trial was stopped early based on a planned interim analysis after half the cohort completed 5 years of follow-up. The median length of follow-up was 4 years (range 3-6 years). Predictors of cognitive change were examined using cross sectional (baseline) and longitudinal multivariable linear regression. The mean (SD) change from baseline to 5 years was -1.9 (5.1) for the SCOPA-COG and -2.1 (11.1) for the SDMT. Age and baseline UPDRS motor scores were associated with a more rapid decline in SDMT scores and 5 year SCOPA-COG scores. Male gender was associated with more rapid decline in SDMT. Self-reported income was a novel predictor of baseline cognitive function, even adjusted for educational status, although not significantly associated with change over time. This large prospective cohort study demonstrated mild cognitive decline in early treated Parkinson's disease. The study identified income level as a novel predictor of cognitive function.


RaPiDS: an algorithm for rapid expression profile database search.
In this paper we present a fast algorithm and implementation for computing the Spearman rank correlation (SRC) between a query expression profile and each expression profile in a database of profiles. The algorithm is linear in the size of the profile database with a very small constant factor. It is designed to efficiently handle multiple profile platforms and missing values. We show that our specialized algorithm and C++ implementation can achieve an approximately 100-fold speed-up over a reasonable baseline implementation using Perl hash tables. RaPiDS is designed for general similarity search rather than classification - but in order to attempt to classify the usefulness of SRC as a similarity measure we investigate the usefulness of this program as a classifier for classifying normal human cell types based on gene expression. Specifically we use the k nearest neighbor classifier with a t statistic derived from SRC as the similarity measure for profile pairs. We estimate the accuracy using a jackknife test on the microarray data with manually checked cell type annotation. Preliminary results suggest the measure is useful (64% accuracy on 1,685 profiles vs. the majority class classifier's 17.5%) for profiles measured under similar conditions (same laboratory and chip platform); but requires improvement when comparing profiles from different experimental series.


Historical perspectives in the care of patients with enterocutaneous fistula.
Evidence can be found throughout surgical history of how devastating an enterocutaneous fistula (ECF) can be for both patient and surgeon. From antiquity, this complication of abdominal surgery, malignancy, radiation, trauma, or inflammatory processes has been a significant challenge to surgeons due to high associated mortality and significant morbidity. An ECF causes dehydration, malnutrition, skin excoriation, and sepsis, and has profound psychological effects on the patient. Recent mortality rates of patients suffering an ECF approach 20%. The authors illustrate the history of management of patients with ECF and discuss advances in perioperative care including parasurgical care, nutrition, wound care, and the history of surgical techniques.


Cryobacterium melibiosiphilum sp. nov., a psychrophilic bacterium isolated from glacier ice.
A psychrophilic, Gram-stain-positive, rod-shaped bacterium, designated Hh39T, was isolated from Xinjiang No. 1 glacier in PR China. Strain Hh39T was catalase-positive, oxidase-negative and could grow at 0-18 °C, pH 6.0-11.0 and in the presence of 0-2.5 % (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain Hh39T belonged to the genus Cryobacterium. The highest level of 16S rRNA gene sequence similarities were found to the type strains of Cryobacterium levicorallinum (99.01 %), Cryobacterium luteum (98.90 %), Cryobacterium aureum (98.90 %) and Cryobacterium roopkundense (98.75 %). However, the low average nucleotide identity (80.65-81.89 %) and digital DNA-DNA hybridization values (22.1-23.8 %) between strain Hh39T and its four closest relatives indicated that it represents a novel species of the genus Cryobacterium. The predominant fatty acids were anteiso-C15:0, anteiso-C15:1, iso-C16:0 and anteiso-C17:0. The major menaquinone was MK-10. The polar lipids were diphosphatidylglycerol, phosphatidylglycerol, one unidentified lipid and one unidentified glycolipid. On the basis of results of phenotypic, genotypic and phylogenetic analyses, a novel species, Cryobacterium melibiosiphilum sp. nov., is proposed, with Hh39T (=NBRC 107884T=CGMCC 1.11212T) as the type strain.


Comparison of loading rate-dependent injury modes in a murine model of post-traumatic osteoarthritis.
Post-traumatic osteoarthritis (PTOA) is a common long-term consequence of joint injuries such as anterior cruciate ligament (ACL) rupture. In this study we used a tibial compression overload mouse model to compare knee injury induced at low speed (1 mm/s), which creates an avulsion fracture, to injury induced at high speed (500 mm/s), which induces midsubstance tear of the ACL. Mice were sacrificed at 0 days, 10 days, 12 weeks, or 16 weeks post-injury, and joints were analyzed with micro-computed tomography, whole joint histology, and biomechanical laxity testing. Knee injury with both injury modes caused considerable trabecular bone loss by 10 days post-injury, with the Low Speed Injury group (avulsion) exhibiting a greater amount of bone loss than the High Speed Injury group (midsubstance tear). Immediately after injury, both injury modes resulted in greater than twofold increases in total AP joint laxity relative to control knees. By 12 and 16 weeks post-injury, total AP laxity was restored to uninjured control values, possibly due to knee stabilization via osteophyte formation. This model presents an opportunity to explore fundamental questions regarding the role of bone turnover in PTOA, and the findings of this study support a biomechanical mechanism of osteophyte formation following injury.


MERS-CoV pathogenesis and antiviral efficacy of licensed drugs in human monocyte-derived antigen-presenting cells.
Middle East respiratory syndrome coronavirus (MERS-CoV) presents an emerging threat to public health worldwide by causing severe respiratory disease in humans with high virulence and case fatality rate (about 35%) since 2012. Little is known about the pathogenesis and innate antiviral response in primary human monocyte-derived macrophages (MDMs) and dendritic cells (MDDCs) upon MERS-CoV infection. In this study, we assessed MERS-CoV replication as well as induction of inflammatory cytokines and chemokines in MDMs and immature and mature MDDCs. Immature MDDCs and MDMs were permissive for MERS-CoV infection, while mature MDDCs were not, with stimulation of proinflammatory cytokine and chemokine upregulation in MDMs, but not in MDDCs. To further evaluate the antiviral activity of well-defined drugs in primary antigen presenting cells (APCs), three compounds (chloroquine, chlorpromazine and toremifine), each with broad-spectrum antiviral activity in immortalized cell lines, were evaluated in MDMs and MDDCs to determine their antiviral effect on MERS-CoV infection. While chloroquine was not active in these primary cells, chlorpromazine showed strong anti-MERS-CoV activity, but it was associated with high cytotoxicity narrowing the potential window for drug utilization. Unlike in established cells, toremifene had marginal activity when tested in antigen presenting cells, with high apparent cytotoxicity, also limiting its potential as a therapeutic option. These results demonstrate the value of testing drugs in primary cells, in addition to established cell lines, before initiating preclinical or clinical studies for MERS treatment and the importance of carefully assessing cytotoxicity in drug screen assays. Furthermore, these studies also highlight the role of APCs in stimulating a robust protective immune response to MERS-CoV infection.


Effect of a simulated soccer match on the functional hamstrings-to-quadriceps ratio in amateur female players.
The purpose of this study was to investigate the effect of a simulated soccer test on the functional hamstrings-to-quadriceps ratio (H(ecc) : Q(con)) in female soccer players. Fourteen amateur players (age, 26.1 ± 4.6 years; height, 168 ± 12 cm; body mass, 62.7 ± 5.5 kg; body fat, 23.7 ± 2.2%) performed the modified Loughborough Intermittent Shuttle Test (modified LIST). Isokinetic strength assessments of the hamstrings and quadriceps on the dominant and non-dominant legs at 120°/s were performed before and immediately after the modified LIST. H(ecc) : Q(con) was calculated as the ratio of the peak eccentric torque of the hamstrings to the peak concentric torque of the quadriceps. A two-way univariate analysis of variance was used to assess the effect of time and leg dominance on H(ecc) : Q(con). The main results showed that the modified LIST led to a significant decrease in H(ecc) : Q(con) in the dominant (-14.1%) and non-dominant legs (-8.0%) (P = 0.02). However, this decrease was not significantly different between dominant and non-dominant legs (P = 0.42). These results reflect a greater risk of hamstrings tears and ACL sprains at the end of soccer matches.


Safety evaluation for a biodiesel process using prion-contaminated animal fat as a source.
Due to the bovine spongiform encephalopathy (BSE), specified risk material (SRM) as well as animal meat and bone meal (MBM) are banned from the food and feed chain because of a possible infection with pathogenic prions (PrP(Sc)). Nowadays, prions are widely accepted to be responsible for TSE(transmissible spongiform encephalopathies)-caused illnesses like BSE and scrapie, and especially for the occurrence of the new variant of CJD in humans. Presently, SRM and MBM are burnt under high temperatures to avoid any hazards for humans, animals or the environment. The aim of this study was to evaluate a method using animal fat separated from Category I material which includes SRM and the carcasses of TSE-infected animals, or animals suspected of being infected with TSE, as a source for producing biodiesel by transesterification, analogous to the biodiesel process using vegetable oil. For this purpose, animal fat was spiked with scrapie-infected hamster brain equivalents--as representative for a TSE-infected animal--and the biodiesel manufacturing process was downscaled and performed under lab-scale conditions. The results analysed by Western blotting showed clearly that almost each single step of the process leads to a significant reduction of the concentration of the pathogenic prion protein (PrP(Sc)) in the main and side-products. The data revealed that the biodiesel production, even from material with a high concentration of pathogenic prions, can be considered as safe. The obtained results indicated that biodiesel produced from prion-contaminated fat was safe under the tested process conditions. However, it has to be pointed out that the results cannot be generalized because a different process control using other conditions may lead to different results and then has to be analysed independently. It is clear that the production of biodiesel from high risk material represents a more economic usage than the combustion of such material.


New genotype of avian influenza H5N1 viruses isolated from tree sparrows in China.
The 2004 outbreaks of highly pathogenic avian influenza H5N1 disease in China led to a great poultry loss and society attention. A survey of avian influenza viruses was conducted on tree sparrows (Passer montanus) collected in China in 2004. Four viruses were isolated from free-living tree sparrows. The results of the whole-genome analysis indicated that an H5N1 virus with a new genotype is circulating among tree sparrows. The hemagglutinin and neuraminidase genes of the new genotype were derived from Gs/Gd/96-like viruses and the nuclear protein gene descended from the 2001 genotype A H5N1 viruses, while the other inner genes originated from an unknown influenza virus. In experimental infection, all four viruses were highly pathogenic to chickens but not pathogenic to ducks or mice. The four tree sparrow viruses were different from the 2003 tree sparrow strain (genotype Z) in Hong Kong. The results suggested that H5N1 viruses might be distributed widely in tree sparrows.


Toward the evaluation of family practice: development of a family utilization index.
This paper describes a research project that establishes criteria for utilization patterns of a family practice that could be used to develop evaluation techniques for family practice as a method of delivering primary medical care. The criteria are summarized in a Family Utilization Index, which measures the utilization patterns of a family unit. Changes in the Utilization Index are compared over a five-year period. Changes in attitudes and opinions about family practice in general and the specific Family Medicine Clinic are also compared over time and related to changes in the Family Utilization Index. Utilization patterns have significantly shifted over a five-year period, with more total visits to the clinic due to an increase in the number of family members per family unit using the clinic. Both the utilization patterns and attitudes toward the role of the family physician have shifted, indicating an increased acceptance of family practice.


Nonnasopharyngeal lymphoepithelioma of the head and neck.
Lymphoepithelioma (squamous cell carcinoma with associated lymphoid stroma) commonly occurs in the nasopharynx, rarely at other sites. As a result, the clinical course and optimal treatment of nonnasopharyngeal lymphoepithelioma of the head and neck have not been well described. This retrospective study was undertaken to analyze the clinical course of the disease in patients treated at a single institution and to formulate recommendations for treatment based on that experience as well as results reported in the literature. Between 1950 and 1994, 34 patients with nonnasopharyngeal lymphoepithelioma of the head and neck were treated at the University of Texas M. D. Anderson Cancer Center. The patients' medical records were reviewed and their pathologic specimens evaluated. The primary tumor sites were: oropharynx (24 patients), salivary gland (4), laryngohypopharynx (4), and the maxillary sinus/nasal cavity (2). Assessed in accordance with the 1992 American Joint Committee Against Cancer TNM staging system, T classifications were TX-2, T1-7, T2-8, T3-10, and T4-7, and N classifications were N0-8, N1-5, N2-15, and N3-6. Treatment consisted of radiotherapy for 24 patients, excisional biopsy of the primary tumor followed by radiotherapy for 7 patients, and surgery for 3 patients. Of the patients treated with radiotherapy, neck dissections were performed on only two, both of whom had persistent lymph node masses after completing radiotherapy. The median dose delivered to the primary tumor was 65 gray (Gy) (range, 46-78 Gy). The median fraction size was 2.1 Gy (range, 1.6-3.2 Gy). The 5-year actuarial disease specific survival and overall survival rates were 59% and 39%, respectively. The 5-year actuarial local control rate for all patients was 94%. For the irradiated patients, the 5-year regional control rates were 77% overall and 83% within the radiation field. The 5-year actuarial rate of distant metastasis for all patients was 30%. For patients who presented with and without regional adenopathy, the 5-year rates of distant metastasis were 36% and 12%, respectively (P = 0.27). Nonnasopharygeal lymphoepithelioma is a radiosensitive disease. High rates of locoregional tumor control were achieved with radiotherapy at all head and neck sites. The main cause of treatment failure was distant metastasis, which occurred more frequently in patients with lymph node involvement. Radiotherapy is appropriate initial locoregional therapy for patients with this disease. Surgery should be reserved for patients who have persistent disease after completing radiotherapy. Systemic therapy is a reasonable approach for patients who present with regional adenopathy because they have a relatively high rate of distant metastasis.


Extension of platelet concentrate storage by addition of sodium bicarbonate.
Viability of platelet concentrate (PC) stored in polyvinylchloride bags in an elliptical rotator at 22 degrees C (standard PC) was assessed by in vitro tests, and an alternate approach to extending the shelf-life of PC by the addition of hypertonic sodium bicarbonate (test PC) was investigated. The fall in the pH which occurred during storage in standard PC was arrested in test PC. Furthermore, platelets stored under these test conditions maintained their morphology better than in standard PC as judged by their mean platelet volume and platelet distribution width. Recovery of stored platelets from hypotonic shock at 37 degrees C following resuspension in fresh plasma was better for test platelets. Results indicated that platelets in standard PC were viable up to day 3 but were not viable at day 7. Platelets store better in PC to which sodium bicarbonate has been added and behave as viable platelets up to 7 days.


Extracellular hydrolytic enzymes produced by yeasts from Antarctic lichens.
In the Antarctic environment, yeasts are versatile eukaryotes that have shown wide dispersion in different substrates, producing active enzymes in extreme conditions, but their relevance in biotechnological applications is largely unknown. The aim of this study was to evaluate the production of extracellular hydrolases by yeasts isolated from Antarctic lichens and molecularly identify these isolates. From a total of 144 isolates on the screening, 109 (76%) produced at least one of the hydrolases tested, with most activities for proteases 59 (41%), cellulases 58 (40%), esterases 57 (39%), lipases 29 (20%), amylases 23 (16%) and pectinases 20 (14%). Among these isolates, 76 were identified, most belonged to the phylum Basidiomycota (n=73) with the dominance of Vishniacozyma victoriae (n=27), Cystobasidium alpinum (n=3), Mrakia niccombsii (n=3), Cystobasidium laryngis (n=2), Bannozyma yamatoana (n=2), Holtermanniella nyarrowii (n=2), and Glaciozyma martinii (n=2). This study is the first one reporting extracellular enzyme production by yeasts isolated from thallus of the species of Antarctic lichens Lecania brialmontii, Polycauliona candelaria, Usnea capillacea, Cladonia metacorallifera, and Polycauliona regalis. With these data, it's possible to confirm lichens as a source of hydrolase-producing yeasts, reinforcing the potential of these microorganisms in bioprospecting studies of catalytic molecules from polar regions that may be useful in promising biotechnological applications.


High resolution 31P NMR of extracted phospholipids.
The common phospholipids from biological sources were quantitated using phosphorus-31 nuclear magnetic resonance (NMR) spectroscopy in conjunction with an analytical reagent composed of two parts: 1) 2 ml of reagent chloroform in which was dissolved 0.01-100 mg of crude tissue lipid extracted from tissue sources using chloroform-methanol 2:1, the extract having been washed with 0.2 vol. of 0.1 M KCl; 2) 1 ml of an aqueous methanol reagent composed of one part 0.2 M (ethylenedinitrilo)-tetraacetic acid in D2O titrated to pH 6 with CsOH and four parts of reagent methanol. In a magnetic field of 11.75 Tesla, the extracted phospholipids yield narrow signals (1.8-3.2 Hz at half-height), corresponding to each generic species, e.g., phosphatidylcholines, phosphatidylethanolamines, etc., permitting resolution among the various phospholipid families and their lyso and plasmalogen derivatives. The reagent permits assays of high precision and accuracy using a modest amount of NMR spectrometer time (ca. 15 min/assay). The procedures described, which are compared to high-performance liquid chromatography, are convenient for the routine analysis of phospholipids from biological sources.


Nutrition in acute pancreatitis.
This review aims to discuss recent developments in different topics regarding nutrition and acute pancreatitis (AP), including oral refeeding, nutritional therapy, and implications of gut microbiota. Obesity increases the risk for severe AP and mortality. Considering the worldwide obesity rates, this finding could have major implications in the global outcomes of patients admitted with AP. Recent research confirms that early oral feeding leads to shorter length of stay, fewer complications, and lower costs. In case of intolerance to oral feeding or severe disease, nutritional therapy should be offered within 24-72 h, whereas enteral nutrition (EN) has been shown superior to parenteral nutrition. EN can be administered through gastric or jejunal feeding, depending on digestive tolerance and the presence of ileus. Nevertheless, modalities of EN in patients undergoing endoscopic drainage of pancreatitis-related collections are still undetermined. Weight-loss after discharge occurs frequently and could reflect post-AP pancreatic exocrine failure. Finally, novel research regarding gut microbiota could open new therapeutic opportunities to prevent bacterial translocation and pancreatic necrosis' infection. Despite available evidence many questions regarding nutritional management in patients with AP remain open. Modulation of gut microbiota could play an important role in further therapeutic management.


Fecal Contamination on the Household Compound and in Water Sources are Associated with Subsequent Diarrhea in Young Children in Urban Bangladesh (CHoBI7 Program).
We investigated the environmental and individual-level risk factors for diarrheal disease among young children in slum areas of Dhaka, Bangladesh. A prospective cohort study was conducted among 884 children under 5 years of age. Caregiver reports were collected on sociodemographic factors and hygiene behaviors. Diarrhea surveillance data was collected monthly based on caregiver-reported diarrhea for children in the past 2 weeks during the 12-month study period. Unannounced spot checks of the household compound were performed at 1, 3, 6, 9, and 12 months after enrollment to check for the presence of feces (animal or human) and the presence of animals in the child's sleeping space, to assess child and caregiver hands for the presence of dirt, and to collect samples of the household's source and stored drinking water. Children with feces found on the household compound during spot checks had a significantly higher odds of diarrhea (odds ratio: 1.71; 95% confidence interval: 1.23-2.38). Children residing in households with > 100 colony forming units/100 mL Escherichia coli in source drinking water had a significantly higher odds of diarrhea (OR: 1.43; 95% CI: 1.06-1.92). The presence of feces on the household compound and source drinking water with > 100 colony forming units/100 mL E. coli were significant risk factors for diarrheal disease for children < 5 years of age in slum areas of Dhaka, Bangladesh. These findings demonstrate the urgent need for comprehensive interventions to reduce fecal contamination on the household compound to protect the health of susceptible pediatric populations.


The Fish Pathogen Vibrio ordalii Under Iron Deprivation Produces the Siderophore Piscibactin.
Vibrio ordalii is the causative agent of vibriosis, mainly in salmonid fishes, and its virulence mechanisms are still not completely understood. In previous works we demonstrated that V. ordalii possess several iron uptake mechanisms based on heme utilization and siderophore production. The aim of the present work was to confirm the production and utilization of piscibactin as a siderophore by V. ordalii. Using genetic analysis, identification by peptide mass fingerprinting (PMF) of iron-regulated membrane proteins and chemical identification by LC-HRMS, we were able to clearly demonstrate that V. ordalii produces piscibactin under iron limitation. The synthesis and transport of this siderophore is encoded by a chromosomal gene cluster homologous to another one described in V. anguillarum, which also encodes the synthesis of piscibactin. Using β-galactosidase assays we were able to show that two potential promoters regulated by iron control the transcription of this gene cluster in V. ordalii. Moreover, biosynthetic and transport proteins corresponding to piscibactin synthesis and uptake could be identified in membrane fractions of V. ordalii cells grown under iron limitation. The synthesis of piscibactin was previously reported in other fish pathogens like Photobacterium damselae subsp. piscicida and V. anguillarum, which highlights the importance of this siderophore as a key virulence factor in Vibrionaceae bacteria infecting poikilothermic animals.


Antoni Christian Bryk (1820-1881) - professor of forensic medicine at Jagiellonian University (1852-1860), and director of the surgical clinic of Jagiellonian University in Cracow (1860-1881).
scientific discipline. In the 19th century two milestones revolutionised surgery: the development of narcosis, which enabled painless surgery, and the introduction of antisepsis and asepsis. The author presents the beginnings of academic surgery in Cracow. Its pioneer surgeons are presented. Extensive research was undertaken to collect the literature and documents in Polish, Austrian and German archives and libraries in order to prepare this study. Biographical details of the director of the Surgical Clinic, Prof. Antoni Bryk, are provided. He was the first person in Poland to introduce antisepsis and galvanocautery as routine procedures in the Cracow Clinic. The introduction of antisepsis contributed to a reduction in infection during surgery, and a reduction in postoperative mortality in Cracow Surgical Clinic. In this way Professor Bryk became the first Polish surgeon to apply Lister's antiseptic method in the treatment of wounds. Thus enlarged, the scope of surgery for intracranial, bone and other procedures became routine. Surgery is the oldest discipline in medicine. Poland's first university chair of surgery was established in the 18th century. Surgery, which until then had been the domain of barbers and bath house attendants, became a clinical,


Current burden of serious fungal infections in Republic of Congo.
The Republic of Congo (RoC) is characterised by a high prevalence of tuberculosis and HIV/AIDS, which largely drive the epidemiology of serious fungal infections. We aimed to estimate the current burden of serious fungal infections in RoC. Using local, regional or global data and estimates of population and at-risk population groups, deterministic modelling was employed to estimate national incidence or prevalence of the most serious fungal infections. Our study revealed that about 5.4% of the Congolese population (283 450) suffer from serious fungal infections yearly. The incidence of cryptococcal meningitis, Pneumocystis jirovecii pneumonia and disseminated histoplasmosis in AIDS patients was estimated at 560, 830 and 120 cases per year. Oral and oesophageal candidiasis collectively affects 12 320 HIV-infected patients. Chronic pulmonary aspergillosis, 67% post-tuberculosis, probably has a prevalence of 3420. Fungal asthma (allergic bronchopulmonary aspergillosis and severe asthma with fungal sensitisation) probably has a prevalence of 3640 and 4800, although some overlap due to disease definition is likely. The estimated prevalence of recurrent vulvovaginal candidiasis and tinea capitis is 85 440 and 178 400 respectively. Mostly related to agricultural activity, fungal keratitis affects an estimated 700 Congolese yearly. These data underline the urgent need for an intensified awareness towards Congolese physicians to fungal infections and for increased efforts to improve diagnosis and management of fungal infections in the RoC.


[Early ventricular repolarization syndrome in patients with heart defects].
The prevalence of the early ventricular repolarization syndrome (EVRS) was examined in 129 patients with congenital and acquired heart diseases. Dynamic ECG values were assessed. The most prevalence rate was found in patients with congenital heart disease (100%), aortic (80%), mitral (51%), and aorto-mitral valvular (60%) diseases. The prevalence of EVRS, the mechanisms of its occurrence were associated with the performance of accessory pathways and the manifestation of the competitive nature of the functional interaction between the basic and accessory pathways.


Determination of amprolium in egg yolk and muscle tissue (chicken) by HPLC with post-column reaction and fluorometric detection, using on-line sample clean-up and pre-concentration steps.
A continuous flow system was coupled to a high-pressure liquid chromatography (HPLC) system, resulting in an automated system for the determination of amprolium in egg yolk and (chicken) muscle tissue. The sample was diluted (yolk) or extracted (tissue) with water, and the solution obtained was dialysed against water as the recipient stream. Aliquots of the dialysed solutions were pumped onto a short pre-concentration column. By means of the mobile phase, the concentrate was back-flushed onto the analytical column and amprolium was separated from interfering substances, using a reversed phase ion-pair system. Amprolium was post-column oxidized to amprochrome, which was detected fluorometrically. Linear calibration curves for both yolk an muscle tissue were obtained in the 10-250 micrograms/kg range. The detection limit is approximately 3 micrograms/kg. This method was applied to eggs and muscle tissue, which were commercial obtained. Egg yolk was found to be frequently contaminated with low levels of amprolium (29.4% positive of 266 samples investigated; mean concentration of positive samples = 58 micrograms/kg), whereas only a few muscle samples contained detectable levels (4.9% positive of 81 samples investigated; mean concentration of positive samples = 5 micrograms/kg).


Comparison of antifungal efficacy of commercially available denture cleanser and the extract of Turbinaria conoides seaweed against Candida albicans adherent to acrylic denture base resin: An in vitro study.
One of the main factors responsible for the development of Candida albicans on the surface of denture is improper maintenance of dentures. Denture hygiene can be achieved by regular cleansing of dentures using an appropriate denture cleanser. The aim of the study is to evaluate the antifungal efficacy of commercially available denture cleanser and the extract of Turbinaria conoides seaweed against C. albicans adherent to the surface of denture base resin. This was an in vitro experimental study. Twenty-four Acrylic resin samples of dimension 10-mm radius and 2-mm thickness was randomly divided into two groups. The denture base resin was coated with C. albicans. The colonies present on the surface of each denture base resin were evaluated by serial dilution method. Group A was treated with commercially available denture cleanser and Group B was treated with extract of seaweed T. conoides. The colonies were then evaluated using serial dilution. The colony count values obtained by serial dilution were tabulated. These values were statistically analysed using t-test. Reduction of colony count is more in T. conoides than commercially available Fittydent; the difference was statistically significant with a mean difference of 65 at dilution 10-2 and 29.25 at dilution 10-3 using t-test with P < 0.001. Within the limitations of this in vitro study, it was proved that the extract of T. conoides seaweed and commercially available denture cleanser Fittydent was effective in reducing the colony count of C. albicans. T. conoides seaweed is statistically significant than commercially available Fittydent.