VapC toxins from Mycobacterium tuberculosis are ribonucleases that differentially inhibit growth and are neutralized by cognate VapB antitoxins.
The chromosome of Mycobacterium tuberculosis (Mtb) encodes forty seven toxin-antitoxin modules belonging to the VapBC family. The role of these modules in the physiology of Mtb and the function(s) served by their expansion are unknown. We investigated ten vapBC modules from Mtb and the single vapBC from M. smegmatis. Of the Mtb vapCs assessed, only Rv0549c, Rv0595c, Rv2549c and Rv2829c were toxic when expressed from a tetracycline-regulated promoter in M. smegmatis. The same genes displayed toxicity when conditionally expressed in Mtb. Toxicity of Rv2549c in M. smegmatis correlated with the level of protein expressed, suggesting that the VapC level must exceed a threshold for toxicity to be observed. In addition, the level of Rv2456 protein induced in M. smegmatis was markedly lower than Rv2549c, which may account for the lack of toxicity of this and other VapCs scored as 'non-toxic'. The growth inhibitory effects of toxic VapCs were neutralized by expression of the cognate VapB as part of a vapBC operon or from a different chromosomal locus, while that of non-cognate antitoxins did not. These results demonstrated a specificity of interaction between VapCs and their cognate VapBs, a finding corroborated by yeast two-hybrid analyses. Deletion of selected vapC or vapBC genes did not affect mycobacterial growth in vitro, but rendered the organisms more susceptible to growth inhibition following toxic VapC expression. However, toxicity of 'non-toxic' VapCs was not unveiled in deletion mutant strains, even when the mutation eliminated the corresponding cognate VapB, presumably due to insufficient levels of VapC protein. Together with the ribonuclease (RNase) activity demonstrated for Rv0065 and Rv0617--VapC proteins with similarity to Rv0549c and Rv3320c, respectively--these results suggest that the VapBC family potentially provides an abundant source of RNase activity in Mtb, which may profoundly impact the physiology of the organism.


Cell interactions in the immune response in vitro. 3. Specific collaboration across a cell impermeable membrane.
Tissue cultures with two compartments, separated by a cell impermeable nuclepore membrane (1 micro pore size), were used to investigate the mechanism of T-B lymphocyte cooperation. It was found that collaboration was as effective when the T and B lymphocyte populations were separated by the membrane as when they were mixed together. Critical tests were performed to verify that the membranes used were in fact cell impermeable. The specificity of the augmentation of the B cell response by various T cell populations was investigated. Only the response of B cells reactive to determinants on the same molecule as recognized by the T cells was augmented markedly. Specific activation of thymocytes by antigen was necessary for efficient collaboration across the membrane. The response of both unprimed and hapten-primed spleen cells was augmented by the T cell "factor" although, as expected, hapten-primed cells yielded greater responses. The T cell factor acted as efficiently if T cells were present or absent in the lower chamber. Thus the site of action of the T cell factor was not on other T cells, but was either on macrophages or the B cells themselves. The T cell-specific immunizing factor did not pass through dialysis membranes. The experiments reported here help rule out some of the possible theories of T-B cell collaboration. Clearly T-B cell contact was not necessary for successful cooperation to occur in this system. Possible theoretical interpretations of the results and their bearing on the detailed mechanism of T-B lymphocyte cooperation are discussed.


MiR-155 targets TP53INP1 to regulate liver cancer stem cell acquisition and self-renewal.
In liver cancer, miR-155 up-regulation can regulate cancer-cell invasion. However, whether miR-155 expression is associated with liver cancer stem cells (CSCs) remains unknown. Here, we show that miR-155 expression is up-regulated in tumor spheres. Knock-down of miR-155 resulted in suppression of tumor sphere formation, through a decrease in the proportion of CD90(+) and CD133(+) CSCs and in the expression of Oct4, whereas miR-155 overexpression had the opposite effect. TP53INP1 was determined to be involved in the CSCs-like properties that were regulated by miR-155. Thus, miR-155 may play an important role in promoting the generation of stem cell-like cells and their self-renewal by targeting the gene TP53INP1.


Spirulina improves non-alcoholic steatohepatitis, visceral fat macrophage aggregation, and serum leptin in a mouse model of metabolic syndrome.
Nutritional approaches are sought to overcome the limits of pioglitazone in metabolic syndrome and non-alcoholic fatty liver disease. Spirulina, a filamentous unicellular alga, reduces serum lipids and blood pressure while exerting antioxidant effects. To determine whether Spirulina may impact macrophages infiltrating the visceral fat in obesity characterizing our metabolic syndrome mouse model induced by the subcutaneous injection treatment of monosodium glutamate. Mice were randomized to receive standard food added with 5% Spirulina, 0.02% pioglitazone, or neither. We tested multiple biochemistry and histology (both liver and visceral fat) readouts at 24 weeks of age. Data demonstrate that both the Spirulina and the pioglitazone groups had significantly lower serum cholesterol and triglyceride levels and liver non-esterified fatty acid compared to untreated mice. Spirulina and pioglitazone were associated with significantly lower leptin and higher levels, respectively, compared to the control group. At liver histology, non-alcoholic fatty liver disease activity score and lipid peroxide were significantly lower in mice treated with Spirulina. Spirulina reduces dyslipidaemia in our metabolic syndrome model while ameliorating visceral adipose tissue macrophages. Human studies are needed to determine whether this safe supplement could prove beneficial in patients with metabolic syndrome.


Long AT-rich palindromes and the constitutional t(11;22) breakpoint.
The constitutional t(11;22) is the most frequently occurring non-Robertsonian translocation in humans. The breakpoint (BP) of the t(11;22) has been identified within palindromic AT-rich repeats (PATRRs) on chromosomes 11 and 22, suggesting that hairpin/cruciform structures mediate double-strand breaks leading to the translocation. To further characterize the mechanism of the translocation, identification of the precise location of the translocation BP is essential. Thus, the PATRRs from normal chromosomes 11 have been analyzed in detail. The majority of individuals have a PATRR that is 445 bp in length with a nearly symmetrical structure. The shorter, previously reported 204 bp PATRR has been shown to be a rare polymorphism. There are several nucleotide differences between the proximal and distal arms of the 445 bp palindrome (cis-morphisms) that correspond to five polymorphic sites within the PATRR. Using these data, the junction fragments of 40 unrelated t(11;22) families have been examined to determine the position of their 11q23 BPs. Sequence analysis demonstrates that BPs are located at the center of the longer PATRR in 39 of 40 cases. The data suggest that the center of the palindrome is susceptible to double-strand breaks leading to translocations that sustain small symmetrical deletions at the BP junction. The sequence of the larger, chromosome 22 PATRR deduced from junction fragments has three cis-morphisms, and the derivative chromosomes sustain symmetric deletions at the center of 22q11 PATRR. In one unusual case, the BPs on both chromosomes appear to correspond to these cis-morphic sites, suggesting that double-strand breaks at mismatched regions caused this variant translocation. De novo t(11;22) BPs have been analyzed using translocations detected in sperm samples from normal males. cis-Morphisms reveal no exclusive utilization of a particular allele in meiosis to produce the translocation. Our data lend support to the hypothesis that palindrome-mediated double-strand breaks in meiosis cause illegitimate recombination between 11q23 and 22q11 resulting in this recurrent translocation.


Induction by monoclonal anti-idiotypic antibodies of an anti-poly(Glu60 Ala30 Tyr10) (GAT) immune response in GAT-responder and GAT-nonresponder mice.
Two different monoclonal anti-idiotypic (Id) antibodies, HP-Id20 and HP-Id22, recognizing two discrete idiotopes characteristic of the anti-poly(Glu60 Ala30 Tyr10) (GAT) response were used to immunize BALB/c (GAT-responder) and DBA/1 (GAT-nonresponder) mice. The monoclonals were injected either copolymerized with keyhole limpet haemocyanin or polymerized with glutaraldehyde. The specific response was studied by two assays: (a) inhibition of binding of monoclonal anti-GAT antibody G5Bb2-2 to HP-Id20 and HP-Id22 and (b) GAT binding assays. In BALB/c GAT-responder mice, HP-Id20 and HP-Id22 immunization led to the preferential stimulation of immunoglobulin idiotypically related to anti-GAT antibodies (Ab1') and expressing anti-GAT activity. The results obtained with BALB/c nu/nu mice indicated that this response is T-cell-dependent. By means of the same experimental protocol GAT-nonresponder animals could be induced to produce anti-GAT antibodies after HP-Id immunization. This last result indicates that anti-Id immunization can bypass Ir gene control and does not preferentially stimulate the induction of GAT-specific T suppressor cells.


Brief communication: Evolution of a specific O allele (O1vG542A) supports unique ancestry of Native Americans.
In this study, we explore the geographic and temporal distribution of a unique variant of the O blood group allele called O1v(G542A) , which has been shown to be shared among Native Americans but is rare in other populations. O1v(G542A) was previously reported in Native American populations in Mesoamerica and South America, and has been proposed as an ancestry informative marker. We investigated whether this allele is also found in the Tlingit and Haida, two contemporary indigenous populations from Alaska, and a pre-Columbian population from California. If O1v(G542A) is present in Na-Dene speakers (i.e., Tlingits), it would indicate that Na-Dene speaking groups share close ancestry with other Native American groups and support a Beringian origin of the allele, consistent with the Beringian Incubation Model. If O1v(G542A) is found in pre-Columbian populations, it would further support a Beringian origin of the allele, rather than a more recent introduction of the allele into the Americas via gene flow from one or more populations which have admixed with Native Americans over the past five centuries. We identified this allele in one Na-Dene population at a frequency of 0.11, and one ancient California population at a frequency of 0.20. Our results support a Beringian origin of O1v(G542A) , which is distributed today among all Native American groups that have been genotyped in appreciable numbers at this locus. This result is consistent with the hypothesis that Na-Dene and other Native American populations primarily derive their ancestry from a single source population.


Peripheral Nerve Blockade for Primary Total Knee Arthroplasty: A Population-based Cohort Study of Outcomes and Resource Utilization.
Although peripheral nerve blocks decrease pain after total knee arthroplasty, the population-level impact of nerve blocks on arthroplasty resource utilization is unknown. We conducted a population-based cohort study using linked administrative data from Ontario, Canada. We identified all adults having their first primary knee arthroplasty between 2002 and 2013. Using propensity scores to adjust for measureable confounders, we matched nerve block patients to a patient who did not receive a block. Within the matched cohort, we estimated the independent association of blocks with outcomes (length of hospital stay [primary]; and readmissions, emergency department visits, and falls [secondary]). One hundred seventy-eight thousand two hundred fourteen patients were identified; 61,588 (34.6%) had a block. The mean hospital stay was 4.6 days with a block compared to 4.8 without. After matching, there was a statistically significant decrease in the length of stay in the block group (relative risk, 0.98; 95% CI, 0.97 to 0.99; P < 0.001). Blocks were associated with a significant decrease in readmissions (relative risk, 0.87; 95% CI, 0.79 to 0.88; P < 0.001) but not emergency department visits (relative risk, 1.02; 95% CI, 0.98 to 1.05) or falls (relative risk, 1.37; 95% CI, 0.90 to 2.08). The association of blocks with length of stay after 2008 was inconsistent; overall, they were associated with longer stays; however, single-shot blocks were associated with shorter stays, while continuous techniques prolonged the length of stay. Nerve blocks in total knee arthroplasty patients were associated with statistically significant reductions in length of stay and readmissions, but not emergency department visits or falls. The significance of these findings at the patient level and in contemporary practice requires further exploration in prospective randomized studies at low risk of indication bias.


Use of intravenous tissue plasminogen activator and hospital costs for patients with acute ischaemic stroke aged 18-64 years in the USA.
Intravenous tissue plasminogen activator (IV tPA) is a globally recommended treatment for acute ischemic stroke patients. We examined IV tPA use among patients aged 18-64 years with a primary diagnosis acute ischemic stroke in the US and inpatient costs per hospitalization by IV tPA use status among these patients. Using 2010-2013 MarketScan Commercial Claims and Encounters Inpatient Data, we identified 39,149 hospitalizations with a primary diagnosis of acute ischemic stroke. We verified those with and without IV tPA by ICD-9 procedure code 99.10. We estimated trends in IV tPA use by applying logistic regression. The average inpatient costs per acute ischemic stroke hospitalization were assessed for sub-populations. We examined costs per acute ischemic stroke hospitalization using multivariate regression models controlling for IV tPA status, age, gender, urbanization, geographic region, Charlson Comorbidity Index, length of hospital stays (LOS) and discharge status. 2,546 hospitalizations (6.5%) used IV tPA. IV tPA use increased over time (2010 vs. 2013; odds ratio 1.50). Average inpatient costs per acute ischemic stroke hospitalization was $20,331 ($31,369 for IV tPA group, $19,563 for non-tPA group). From multivariate analyses, higher costs per acute ischemic stroke hospitalization were associated with longer LOS, non-home discharge destination, and IV tPA use, which might be correlated with severity of stroke. Findings suggest that IV tPA use increased recent years while the inpatient costs per acute ischemic stroke hospitalization using IV tPA are substantial. Those findings are useful in better understanding the overall economic burden of stroke, short-term cost implications of using IV tPA, and for estimating the accurate cost-effectiveness of stroke treatments.


Kinetic path of genes undergoing selection.
As natural populations approach genetic equilibrium, the various genes in the population are capable of assuming intermediate distributions that might not be anticipated from either the rate of the process or the final distribution of the genes. Since it is possible that many populations have not reached genetic equilibrium, the distribution of genes in natural populations may be a reflection of the kinetic path by which the genes approach equilibrium. Attention to kinetic path provides an explanation for an apparent discrepancy in recent studies of selection in man.


A "transmission gap" between research and practice? A Q-methodology study of perceptions of the application of attachment theory among clinicians working with children and among attachment researchers.
Clinical practitioners are frequently encouraged, through literature, training, and policy, to learn, understand, refer to and use their knowledge of attachment theory and research when working to meet the needs of children and families. However, there has been very little empirical study of how practitioners understand and perceive the relevance of attachment concepts and methods. Q-methodology was used to examine the perceptions of attachment knowledge and its applications for practice among 30 UK clinicians working with children and an international group of 31 attachment researchers. Factor analysis revealed three perspectives, described as: i) pragmatic, developmental, and uncertain, ii) academic, and iii) autodidactic and therapeutic. Participants agreed on core tenants of theory, their aspirations for clinical practice and the inaccessibility of current assessment measures for practitioners. Yet they diverged on their understandings of attachment insecurity, disorganisation, and the implications of both for various aspects of child development.


Polarity-specific effects of motor transcranial direct current stimulation on fMRI resting state networks.
Transcranial direct current stimulation (tDCS) has been used to modify motor performance in healthy and patient populations. However, our understanding of the large-scale neuroplastic changes that support such behavioural effects is limited. Here, we used both seed-based and independent component analyses (ICA) approaches to probe tDCS-induced modifications in resting state activity with the aim of establishing the effects of tDCS applied to the primary motor cortex (M1) on both motor and non-motor networks within the brain. Subjects participated in three separate sessions, during which resting fMRI scans were acquired before and after 10min of 1mA anodal, cathodal, or sham tDCS. Cathodal tDCS increased the inter-hemispheric coherence of resting fMRI signal between the left and right supplementary motor area (SMA), and between the left and right hand areas of M1. A similar trend was documented for the premotor cortex (PMC). Increased functional connectivity following cathodal tDCS was apparent within the ICA-generated motor and default mode networks. Additionally, the overall strength of the default mode network was increased. Neither anodal nor sham tDCS produced significant changes in resting state connectivity. This work indicates that cathodal tDCS to M1 affects the motor network at rest. In addition, the effects of cathodal tDCS on the default mode network support the hypothesis that diminished top-down control may contribute to the impaired motor performance induced by cathodal tDCS.


Mechanical analysis of feeding behavior in the extinct "terror bird" Andalgalornis steulleti (Gruiformes: Phorusrhacidae).
The South American phorusrhacid bird radiation comprised at least 18 species of small to gigantic terrestrial predators for which there are no close modern analogs. Here we perform functional analyses of the skull of the medium-sized (approximately 40 kg) patagornithine phorusrhacid Andalgalornis steulleti (upper Miocene-lower Pliocene, Andalgalá Formation, Catamarca, Argentina) to assess its mechanical performance in a comparative context. Based on computed tomographic (CT) scanning and morphological analysis, the skull of Andalgalornis steulleti is interpreted as showing features reflecting loss of intracranial immobility. Discrete anatomical attributes permitting such cranial kinesis are widespread phorusrhacids outgroups, but this is the first clear evidence of loss of cranial kinesis in a gruiform bird and may be among the best documented cases among all birds. This apomorphic loss is interpreted as an adaptation for enhanced craniofacial rigidity, particularly with regard to sagittal loading. We apply a Finite Element approach to a three-dimensional (3D) model of the skull. Based on regression analysis we estimate the bite force of Andalgalornis at the bill tip to be 133 N. Relative to results obtained from Finite Element Analysis of one of its closest living relatives (seriema) and a large predatory bird (eagle), the phorusrhacid's skull shows relatively high stress under lateral loadings, but low stress where force is applied dorsoventrally (sagittally) and in "pullback" simulations. Given the relative weakness of the skull mediolaterally, it seems unlikely that Andalgalornis engaged in potentially risky behaviors that involved subduing large, struggling prey with its beak. We suggest that it either consumed smaller prey that could be killed and consumed more safely (e.g., swallowed whole) or that it used multiple well-targeted sagittal strikes with the beak in a repetitive attack-and-retreat strategy.


The Toxoplasma gondii cyst wall protein CST1 is critical for cyst wall integrity and promotes bradyzoite persistence.
Toxoplasma gondii infects up to one third of the world's population. A key to the success of T. gondii as a parasite is its ability to persist for the life of its host as bradyzoites within tissue cysts. The glycosylated cyst wall is the key structural feature that facilitates persistence and oral transmission of this parasite. Because most of the antibodies and reagents that recognize the cyst wall recognize carbohydrates, identification of the components of the cyst wall has been technically challenging. We have identified CST1 (TGME49_064660) as a 250 kDa SRS (SAG1 related sequence) domain protein with a large mucin-like domain. CST1 is responsible for the Dolichos biflorus Agglutinin (DBA) lectin binding characteristic of T. gondii cysts. Deletion of CST1 results in reduced cyst number and a fragile brain cyst phenotype characterized by a thinning and disruption of the underlying region of the cyst wall. These defects are reversed by complementation of CST1. Additional complementation experiments demonstrate that the CST1-mucin domain is necessary for the formation of a normal cyst wall structure, the ability of the cyst to resist mechanical stress, and binding of DBA to the cyst wall. RNA-seq transcriptome analysis demonstrated dysregulation of bradyzoite genes within the various cst1 mutants. These results indicate that CST1 functions as a key structural component that confers essential sturdiness to the T. gondii tissue cyst critical for persistence of bradyzoite forms.


Diagnostic accuracy of oblique chest radiograph for occult pneumothorax: comparison with ultrasonography.
An occult pneumothorax is a pneumothorax that is not seen on a supine chest X-ray but is detected by computed tomography scanning. However, critical patients are difficult to transport to the computed tomography suite. We previously reported a method to detect occult pneumothorax using oblique chest radiography (OXR). Several authors have also reported that ultrasonography is an effective technique for detecting occult pneumothorax. The aim of this study was to evaluate the usefulness of OXR in the diagnosis of the occult pneumothorax and to compare OXR with ultrasonography. All consecutive blunt chest trauma patients with clinically suspected pneumothorax on arrival at the emergency department were prospectively included at our tertiary-care center. The patients underwent OXR and ultrasonography, and underwent computed tomography scans as the gold standard. Occult pneumothorax size on computed tomography was classified as minuscule, anterior, or anterolateral. One hundred and fifty-nine patients were enrolled. Of the 70 occult pneumothoraces found in the 318 thoraces, 19 were minuscule, 32 were anterior, and 19 were anterolateral. The sensitivity and specificity of OXR for detecting occult pneumothorax was 61.4 % and 99.2 %, respectively. The sensitivity and specificity of lung ultrasonography was 62.9 % and 98.8 %, respectively. Among 27 occult pneumothoraces that could not be detected by OXR, 16 were minuscule and 21 could be conservatively managed without thoracostomy. OXR appears to be as good method as lung ultrasonography in the detection of large occult pneumothorax. In trauma patients who are difficult to transfer to computed tomography scan, OXR may be effective at detecting occult pneumothorax with a risk of progression.


Experimental demonstration of both inverted and non-inverted wavelength conversion based on transient cross phase modulation of SOA.
We demonstrate experimentally ultrafast inverted and noninverted wavelength conversion (WC) based on a semiconductor optical amplifier (SOA) and an optical bandpass filter (OBF). In the case of small detuning, the WC is inverted regardless if the OBF is blue- or red-shifted with respect to the central wavelength of the converted signal. However the WC is non-inverted when the filter detuning is relatively large. An analytical formula for the transient cross phase modulation is applied to reveal the polarity variation of WC with respect to the OBF detuning. The theoretical detuning values are in good agreement with our experimental results.


Anti-hyperalgesic and anti-nociceptive potentials of standardized grape seed proanthocyanidin extract against CCI-induced neuropathic pain in rats.
Neuropathic pain is associated with severe chronic sensory disturbances characterized by spontaneous pain, increased responsiveness to painful stimuli (hyperalgesia) and pain perceived in response to non-noxious stimuli (allodynia). Morphine is effective treatment for neuropathic pain but produces tolerance on chronic use. The present study was designed to explore the anti-nociceptive and anti-hyperalgesic effect of grape seed extract using sciatic nerve ligation-induced neuropathic pain in rats. Chronic constructive injury (CCI) was performed under anesthesia, on one side leg exposed by making a skin incision, and chromic gut ligatures were tied loosely around the sciatic nerve at 1 mm intervals. The treatment with grape seed proanthocyanidin extract (GSPE) (100 and 200 mg/kg, p.o.) was initiated on 7th day post-surgery and continued for next 14 days. Morphine (10 mg/kg, s.c.) alone and morphine in combination with GSPE (100 mg/kg, p.o.) were administered in CCI rats for 5 days starting from 7th day. On 3rd, 7th, 14th and 21st day, behavioral parameters (mechanical allodynia and thermal hyperalgesia) were assessed. Then the animals were killed on 22nd day and biochemical parameters [reduced glutathione (GSH), lipid peroxidation (LPO), catalase, nitrite, superoxide dismutase (SOD)] were assessed. Ligation of the sciatic nerve significantly induced mechanical allodynia and thermal hyperalgesia and induces oxidative stress (increase in LPO and nitrite) and decline of anti-oxidant enzyme levels (catalase, SOD, GSH) in sciatic nerve homogenate. GSPE (100 and 200 mg/kg, p.o.) attenuated all the behavioural and biochemical parameters. Morphine also significantly reversed the symptoms of neuropathic pain but produced tolerance after 5 days. Further, co-treatment of GSPE (100 mg/kg) with morphine (10 mg/kg, s.c.) in CCI rats significantly reversed the morphine tolerance and enhanced its anti-hyperalgesic effect as compared to the morphine-alone-treated group. In the present set of experiments, GSPE showed a significant anti-hyperalgesic and anti-nociceptive effect in rats.


Evaluating the Effectiveness of Social Distancing Interventions to Delay or Flatten the Epidemic Curve of Coronavirus Disease.
By April 2, 2020, >1 million persons worldwide were infected with severe acute respiratory syndrome coronavirus 2. We used a mathematical model to investigate the effectiveness of social distancing interventions in a mid-sized city. Interventions reduced contacts of adults >60 years of age, adults 20-59 years of age, and children <19 years of age for 6 weeks. Our results suggest interventions started earlier in the epidemic delay the epidemic curve and interventions started later flatten the epidemic curve. We noted that, while social distancing interventions were in place, most new cases, hospitalizations, and deaths were averted, even with modest reductions in contact among adults. However, when interventions ended, the epidemic rebounded. Our models suggest that social distancing can provide crucial time to increase healthcare capacity but must occur in conjunction with testing and contact tracing of all suspected cases to mitigate virus transmission.


Pediatrics and donor-derived disease transmission: The US OPTN experience.
PDDTE are tracked by the OPTN Ad Hoc DTAC. Specific evaluation of potential transmissions from pediatric deceased donors or the impact of donor-derived disease transmissions to pediatric organ recipients has not been previously undertaken. PDDTE reported to the DTAC between 2008 and 2013 were reviewed, characterized as proven, probable, possible, IWDT, unlikely, or excluded for both the whole event and each individual recipient. Pediatric donors and recipients were defined as being 0-17 years of age. Analysis was undertaken to characterize potential disease transmission from pediatric donors to adult or pediatric recipients and also to evaluate potential transmission from all donors to pediatric recipients. P/P cases were further analyzed. A total of 5238 pediatric deceased US donors accounted for 17 456 organ transplants during the study period; 103 PDDTE reports arose from these donors (2.0%). PDDTE were characterized as P/P (15%), possible (13%), IWDT (9%), unlikely, and excluded (63%). Disease was transmitted to 22 of 54 potentially exposed (adult and pediatric) recipients with six attributable deaths. An infectious pathogen accounted for 13/15 of the P/P PDDTE associated with pediatric donors, affecting 19 of 50 potentially exposed recipients and resulting in five deaths. Four separate viral pathogens from six donors accounted for P/P transmissions to 11 recipients with the unanticipated transmission of CMV most common. No pediatric donor transmitted HIV, HBV, or HCV. Bacteria, fungi, and parasites accounted for three (all staphylococci), three (Zygomycetes and Histoplasma), and two (both Toxoplasma) P/P transmissions from seven donors, respectively. From the recipient side, 11/11,188 pediatric recipient deceased and living donor transplants during the study period were associated with a P/P PDDTE (<0.1%) with infectious pathogens accounting for 9/11 P/P events. Infections were split among pathogen categories (bacteria 2, viruses 3, parasites 3, and fungi 1). Reporting rates of PDDTE involving pediatric donors were very low and similar to rates from all donors, with resulting P/P transmissions occurring in only 0.1% of exposed recipients, but transmissions were associated with six deaths. Rates of P/P transmission to pediatric recipients from any donor (<0.1%) were also very low and similar to that of all recipients. Additional studies are needed to compare the pattern and outcome of donor-derived disease transmission from and to pediatric and adult donor and recipients.


A molecular survey of biomphalaria in Egypt: is B. glabrata present?
Two species of Biomphalaria are reported from Egypt, the indigenous Biomphalaria alexandrina and Biomphalaria glabrata, the latter believed to be introduced during the past few decades. Both are known to be excellent hosts of Schistosoma mansoni, the human-infecting blood fluke common in Egypt. Given the concerns regarding the spread of the exotic B. glabrata, this study was carried out to get a more current picture of the status of Biomphalaria in Egypt. Snail collections were undertaken during 2002-2003 from regions between Alexandria and Ismailia in the north of the Nile Delta, to as far south as Abu Simbel at Lake Nasser. Biomphalaria snails were found in 37 out of 76 sampled localities and were widely distributed in the Nile Delta and along the Nile as far south as Aswan. According to the results of species-specific polymerase chain reaction assays that sampled both nuclear and mitochondrial genomes, and according to DNA sequence data, all Biomphalaria collected during this survey were B. alexandrina. There was no evidence of the presence of B. glabrata or of hybridization of B. alexandrina with B. glabrata in the examined sites. The results were surprising given that some field-collected snails strongly resembled B. glabrata in both size and conchology and that previous survey work suggested B. glabrata had established in Egypt. Continued scrutiny to ascertain the possible presence of B. glabrata in Egypt is warranted. Also, the planorbid Helisoma duryi was detected in the Delta and as far south as Aswan, so it is important for Egyptian schistosomiasis workers to accurately distinguish this non-schistosome-transmitting snail from Biomphalaria.


Age criteria in medicine. Are the medical justifications ethical?
Age criteria are increasingly being advocated as means of determining who should receive treatment. Four types of medical justification have been proposed. Three of these, namely, length of medical benefit, quality of medical benefit, and likelihood of medical benefit, entail comparisons of patients. The fourth, medical benefit, involves identifying for each patient whether or not a significant benefit is likely to result from treatment. I contend that the three types of comparative justification are questionable. For example, they rest on statistical generalities that are misleading in particular situations, and they value characteristics of persons (such as life-years) more than they respect persons themselves. I also contend that the fourth type of medical justification, rooted as it is in response to human need, is ethically sound, but that it warrants a medical-benefit criterion rather than an age criterion.


Recovery of Campylobacter from commercial broiler hatchery trayliners.
Previous research has identified Campylobacter as one of the leading causes of foodborne illness. Poultry and poultry products have been identified as a major source of Campylobacter in human infections. Although many risk factors that contribute to Campylobacter levels have been identified, precise identification of the most effective sites for intervention has not been established. Epidemiological studies have identified that Campylobacter in the broiler breeder's reproductive tract, fertile eggs, and 2- to 3-wk-old broilers has the potential to contaminate day-of-hatch chicks. Numerous studies have shown that day-of-hatch broilers are Campylobacter-negative using conventional culture methods. The purpose of the present study was to demonstrate the prevalence of Campylobacter found in day-of-hatch broilers using a peptone water preenrichment followed by conventional Campylobacter culture methods. Using conventional tray liner (hatcheries) culture methods, the isolation distribution of Campylobacter from 8 commercial broiler hatcheries (n = 2,000) was evaluated. A total of 15 tray liners were positive from 3 different hatcheries. Of the 2,000 chick paper pad tray liners sampled, 0.75% were positive for Campylobacter. These data support previous findings indicating the potential for Campylobacter to be spread by vertical transmission. This is the first time that Campylobacter has been recovered from tray liners collected at commercial broiler hatcheries.


MAX mutant small-cell lung cancers exhibit impaired activities of MGA-dependent noncanonical polycomb repressive complex.
The MYC axis is disrupted in cancer, predominantly through activation of the MYC family oncogenes but also through inactivation of the MYC partner MAX or of the MAX partner MGA. MGA and MAX are also members of the polycomb repressive complex, ncPRC1.6. Here, we use genetically modified MAX-deficient small-cell lung cancer (SCLC) cells and carry out genome-wide and proteomics analyses to study the tumor suppressor function of MAX. We find that MAX mutant SCLCs have ASCL1 or NEUROD1 or combined ASCL1/NEUROD1 characteristics and lack MYC transcriptional activity. MAX restitution triggers prodifferentiation expression profiles that shift when MAX and oncogenic MYC are coexpressed. Although ncPRC1.6 can be formed, the lack of MAX restricts global MGA occupancy, selectively driving its recruitment toward E2F6-binding motifs. Conversely, MAX restitution enhances MGA occupancy to repress genes involved in different functions, including stem cell and DNA repair/replication. Collectively, these findings reveal that MAX mutant SCLCs have either ASCL1 or NEUROD1 or combined characteristics and are MYC independent and exhibit deficient ncPRC1.6-mediated gene repression.


Computer modeling of 2D supramolecular nanoporous monolayers self-assembled on graphite.
Nano-porous two-dimensional molecular crystals, self-assembled on atomically flat host surfaces offer a broad range of possible applications, from molecular electronics to future nano-machines. Computer-assisted designing of such complex structures requires numerically intensive modeling methods. Here we present the results of extensive, fully atomistic simulations of self-assembled monolayers of interdigitated molecules of 1,3,5-tristyrilbenzene substituted by C6 alkoxy peripheral chains (TSB3,5-C6), deposited onto highly-ordered pyrolytic graphite. Structural and electronic properties of the TSB3,5-C6 molecules were determined from ab initio calculations, then used in Molecular Dynamics simulations to analyze the mechanism of formation, epitaxy, and stability of the TSB3,5-C6 nanoporous superlattice. We show that the monolayer disordering results from the competition between flexibility of the C6 chains and their stabilization by interdigitation. The inclusion of guest molecules (benzene and pyrene) into superlattice nanopores stabilizes the monolayer. The alkoxy chain mobility and available pore space defines the systems dynamics, essential for potential application.


Efficacy of a quadrivalent prophylactic human papillomavirus (types 6, 11, 16, and 18) L1 virus-like-particle vaccine against high-grade vulval and vaginal lesions: a combined analysis of three randomised clinical trials.
Vulval and vaginal cancers among younger women are often related to infection with human papillomavirus (HPV). These cancers are preceded by high-grade vulval intraepithelial neoplasia (VIN2-3) and vaginal intraepithelial neoplasia (VaIN2-3). Our aim was to do a combined analysis of three randomised clinical trials to assess the effect of a prophylactic quadrivalent HPV vaccine on the incidence of these diseases. 18 174 women (16-26 years) were enrolled and randomised to receive either quadrivalent HPV6/11/16/18 L1 virus-like-particle vaccine or placebo at day 1, and months 2 and 6. Individuals underwent detailed anogenital examination at day 1, 1 month after dose three, and at 6-12-month intervals for up to 48 months. Suspect genital lesions were biopsied and read by a panel of pathologists and vaccine HPV type-specific DNA testing was done. The primary endpoint was the combined incidence of VIN2-3 or VaIN2-3 associated with HPV16 or HPV18. Primary efficacy analyses were done in a per-protocol population. The mean follow-up time was 3 years. Among women naive to HPV16 or HPV18 through 1 month after dose three (per-protocol population; vaccine n=7811; placebo n=7785), the vaccine was 100% effective (95% CI 72-100) against VIN2-3 or VaIN2-3 associated with HPV16 or HPV18. In the intention-to-treat population (which included 18 174 women who, at day 1, could have been infected with HPV16 or HPV18), vaccine efficacy against VIN2-3 or VaIN2-3 associated with HPV16 or HPV18 was 71% (37-88). The vaccine was 49% (18-69) effective against all VIN2-3 or VaIN2-3, irrespective of whether or not HPV DNA was detected in the lesion. The most common treatment-related adverse event was injection-site pain. Prophylactic administration of quadrivalent HPV vaccine was effective in preventing high-grade vulval and vaginal lesions associated with HPV16 or HPV18 infection in women who were naive to these types before vaccination. With time, such vaccination could result in reduced rates of HPV-related vulval and vaginal cancers.


Phosphoinositide 3-kinase signaling is critical for ErbB3-driven breast cancer cell motility and metastasis.
Many malignancies show increased expression of the epidermal growth factor (EGF) receptor family member ErbB3 (HER3). ErbB3 binds heregulin β-1 (HRGβ1) and forms a heterodimer with other ErbB family members, such as ErbB2 (HER2) or EGF receptor (EGFR; HER1), enhancing phosphorylation of specific C-terminal tyrosine residues and activation of downstream signaling pathways. ErbB3 contains six YXXM motifs that bind the p85 subunit of phosphoinositide 3 (PI3)-kinase. Previous studies demonstrated that overexpression of ErbB3 in mammary tumor cells can significantly enhance chemotaxis to HRGβ1 and overall metastatic potential. We tested the hypothesis that ErbB3-mediated PI3-kinase signaling is critical for heregulin-induced motility, and therefore crucial for ErbB3-mediated invasion, intravasation and metastasis. The tyrosines in the six YXXM motifs on the ErbB3 C-terminus were replaced with phenylalanine. In contrast to overexpression of the wild-type ErbB3, overexpression of the mutant ErbB3 did not enhance chemotaxis towards HRGβ1 in vitro or in vivo. We also observed reduced tumor cell motility in the primary tumor by multiphoton microscopy, as well as a dramatically reduced ability of these cells to cross the endothelium and intravasate into the circulation. Moreover, whereas mutation of the ErbB3 C-terminus had no effect on tumor growth, it had a dramatic effect on spontaneous metastatic potential. Treatment with the PI3-kinase inhibitor PIK-75 similarly inhibited motility and invasion in vitro and in vivo. Our results indicate that stimulation of the early metastatic steps of motility and invasion by ErbB3 requires activation of the PI3-kinase pathway by the ErbB3 receptor.


The In Vitro Impact of the Herbicide Roundup on Human Sperm Motility and Sperm Mitochondria.
Toxicants, such as herbicides, have been hypothesized to affect sperm parameters. The most common method of exposure to herbicides is through spraying or diet. The aim of the present study was to investigate the effect of direct exposure of sperm to 1 mg/L of the herbicide Roundup on sperm motility and mitochondrial integrity. Sperm samples from 66 healthy men who were seeking semen analysis were investigated after written informed consent was taken. Semen analysis was performed according to the World Health Organization guidelines (WHO, 2010). Mitochondrial integrity was assessed through mitochondrial staining using a mitochondria-specific dye, which is exclusively incorporated into functionally active mitochondria. A quantity of 1 mg/L of Roundup was found to exert a deleterious effect on sperm's progressive motility, after 1 h of incubation (mean difference between treated and control samples = 11.2%) in comparison with the effect after three hours of incubation (mean difference = 6.33%, p < 0.05), while the relative incorporation of the mitochondrial dye in mitochondria of the mid-piece region of Roundup-treated spermatozoa was significantly reduced compared to relative controls at the first hour of incubation, indicating mitochondrial dysfunction by Roundup. Our results indicate that the direct exposure of semen samples to the active constituent of the herbicide Roundup at the relatively low concentration of 1 mg/L has adverse effects on sperm motility, and this may be related to the observed reduction in mitochondrial staining.


Total parenteral nutrition-associated cholestasis and risk factors in preterm infants.
Development of hepatic dysfunction is a well-recognized complication of total parenteral nutrition in preterm infants. Previous studies reported the incidence of total parenteral nutrition-associated cholestasis and described possible contributing factors to its pathogenesis, but little is done trying to determine its possible predictive risk factors. The aims of this study was to determine the incidence of total parenteral nutrition-associated cholestasis and to develop a possible predictive model for its occurrence. A review of medical records of all very low birth weight infants admitted to neonatal intensive care unit at King Khalid University Hospital, Riyadh, Saudi Arabia, between January 2001 and December 2003 was carried out. The infants were divided into two groups: Cholestasis and noncholestasis, based on direct serum bilirubin level >34 μmol/L. A multivariate logistic regression analysis was performed to calculate the statistical significance of risk factors. Receiver-operating characteristic curve was used to determine the optimal cutoff points for the significant risk factors and to calculate their sensitivity and specificity. The level of significance was set at P ≤ 0.05. A total of 307 patients were included in the analysis. The incidence of cholestasis in the whole population was 24.1% (74 patients). Infants with cholestasis had a lower birth weight, 735.4 ± 166.4 g vs. 1185.0 ± 205.6 g for noncholestasis group (P < 0.001), whereas the mean gestational age for the two groups was 25.4 ± 2.1 week and 28.9 ± 2.1 week, respectively (P < 0.001). The significant risk factors for the development of cholestasis were birth weight (P = 0.006) with an odds ratio of 0.99 [95% confidence interval (CI), 0.98, 0.99]; sensitivity of 92%, specificity of 87%; and total parenteral nutrition duration (P < 0.001) with an odds ratio of 1.18 (95% CI, 1.10, 1.27); sensitivity of 96%, specificity of 89%. A lower birth weight and longer duration of total parenteral nutrition were strong predictive risk factors for the development of cholestasis in preterm infants.


A critical interaction between NR2B and MAGUK in L-DOPA induced dyskinesia.
Abnormal function of NMDA receptor has been suggested to be correlated with the pathogenesis of Parkinson's disease (PD) as well as with the development of l-3,4-dihydroxyphenylalanine (L-DOPA)-induced dyskinesia. Here we show that NMDA receptor NR2 subunits display specific alterations of their subcellular distribution in striata from unilateral 6-hydroxydopamine-lesioned, L-DOPA-treated dyskinetic, and L-DOPA-treated nondyskinetic rats. Dyskinetic animals have significantly higher levels of NR2A subunit in the postsynaptic compartment than all other experimental groups, whereas NR2B subunit shows a significant reduction in both dopamine-denervated and dyskinetic rats. These events are paralleled by profound modifications of NMDA receptor NR2B subunit association with interacting elements, i.e., members of the membrane-associated guanylate kinase (MAGUK) protein family postsynaptic density-95, synapse-associated protein-97 and synapse-associated protein-102. Treatment of nondyskinetic animals with a synthetic peptide (TAT2B) able to affect NR2B binding to MAGUK proteins as well as synaptic localization of this subunit in nondyskinetic rats was sufficient to induce a shift of treated rats toward a dyskinetic motor behavior. These data indicate abnormal NR2B redistribution between synaptic and extrasynaptic membranes as an important molecular disturbance of the glutamatergic synapse involved in L-DOPA-induced dyskinesia.


A Smart-Anomaly-Detection System for Industrial Machines Based on Feature Autoencoder and Deep Learning.
Machine-health-surveillance systems are gaining popularity in industrial manufacturing systems due to the widespread availability of low-cost devices, sensors, and internet connectivity. In this regard, artificial intelligence provides valuable assistance in the form of deep learning methods to analyze and process big machine data. In diverse industrial applications, gears are considered a condemning element; many contributing failures occur due to an unexpected breakdown of the gears. In recent research, anomaly-detection and fault-diagnosis systems have been the gears' most contributing content. Thus, in work, we presented a smart deep learning-based system to detect anomalies in an industrial machine. Our system used vibrational analysis methods as a deciding tool for different machinery-maintenance decisions. We will first perform a data analysis of the gearbox data set to analyze the data's insights. By calculating and examining the machine's vibration, we aim to determine the nature and severity of the defect in the machine and hence detect the anomaly. A gearbox's vibration signal holds the fault's signature in the gears, and earlier fault detection of the gearbox is achievable by examining the vibration signal using a deep learning technique. Therefore, we aim to propose a 6-layer autoencoder-based deep learning framework for anomaly detection and fault analysis using a publically available data set of wind-turbine components. The gearbox fault-diagnosis data set is utilized for experimentation, including collecting vibration attributes recorded using SpectraQuest's gearbox fault-diagnostics simulator. Through comprehensive experiments, we have seen that the framework gains good results compared to others, with an overall accuracy of 91%.


Perceived discrimination and psychosis: a systematic review of the literature.
Higher rates of psychosis have been reported in minority groups. Since individuals belonging to such groups are vulnerable to the experiences of discrimination, and in line with models proposing that social and life adversity may play a causal role in development and maintenance of psychotic experiences, it has been proposed that perceived discrimination may represent an important determinant of psychotic experiences. This paper reviews the literature examining the relationship between perceived discrimination and psychosis, examining whether discrimination is associated with an increased risk of psychosis, the severity of psychotic symptoms and whether there is an association with specific psychotic symptoms. A systematic database search of PsycINFO, Embase and PubMed was conducted to identify quantitative cross-sectional and prospective studies that examined the association between discrimination and psychosis. Twenty-four studies met the inclusion criteria, four of which used prospective designs and twenty used cross-sectional designs. The main findings indicated that discrimination may be associated with an increased risk of psychosis (too few studies to determine whether discrimination is associated with severity). Some studies found associations between discrimination and positive psychotic experiences and/or specific psychotic experiences such as paranoia. A small number of studies found that greater exposure to discrimination was associated with a greater likelihood of reporting psychotic experiences, tentatively indicating a dose-response relationship. This review indicates that discrimination plays an important role in the experience of psychosis; however, future research is required to clarify the nature of this relationship. Avenues for further research and clinical implications are proposed.


A prodrug of cysteine, L-2-oxothiazolidine-4-carboxylic acid, regulates vascular permeability by reducing vascular endothelial growth factor expression in asthma.
Inflammation of the asthmatic airway is usually accompanied by increased vascular permeability and plasma exudation. Oxidative stress plays critical roles in airway inflammation. Although reactive oxygen species (ROS) are shown to cause vascular leakage, the mechanisms by which ROS induce increased vascular permeability are not clearly understood. We have used a murine model of asthma to evaluate the effect of l-2-oxothiazolidine-4-carboxylic acid (OTC), a prodrug of cysteine that acts as an antioxidant, more specifically in the increase of vascular permeability. These mice develop the following typical pathophysiological features of asthma in the lungs: increased numbers of inflammatory cells of the airways, airway hyper-responsiveness, increased vascular permeability, and increased levels of vascular endothelial growth factor (VEGF). Administration of OTC markedly reduced plasma extravasation and VEGF levels in allergen-induced asthmatic lungs. We also showed that at 72 h after ovalbumin inhalation, increased levels of hypoxia-inducible factor-1alpha (a transcriptional activator of VEGF) in nuclear protein extracts of lung tissues were decreased by the administration of OTC. These results indicate that OTC modulates vascular permeability by lowering VEGF expression.


Reliability of the Nursing Child Assessment Feeding Scale during toddlerhood.
The quality of the maternal-child feeding interaction has been proposed to be an important contributor to a child's being overweight, yet assessment of this proposition has been hindered by a lack of age-appropriate instrumentation. The primary aim of this study was to examine the reliability of the Nursing Child Assessment of Feeding Scale (NCAFS) if extended to use during toddlerhood. A longitudinal design was used to assess NCAFS reliability at 12, 24, and 36 months. The NCAFS was used to code videotaped feeding observations of 116 mother-toddler dyads collected as part of a larger study examining mother-child interactions and adaptations of toddlers. Reliability was explored through the assessment of interrater reliability, internal consistency of the various subscales and the scale as a whole, and stability of the scale measurements over time. At each age, interrater reliability was generally quite good whereas the NCAFS' internal consistency was low. Maternal contributions to feeding interaction quality were stable over time but dyadic and child contributions were not. The lower internal consistency estimates were likely due to relatively low levels of variance among the dyads within each age. Another probable contributor to diminished internal consistency was the attrition of several behavior items due to zero variance. Possible explanations for this restriction of variance and several approaches for improving the NCAFS internal consistency during toddlerhood are considered. With revision, the NCAFS could be useful in assessment of feeding interaction quality during the transition to toddlerhood when issues of control and autonomy become increasingly prominent.


Nursing management of postoperative pain: use of relaxation techniques with female cholecystectomy patients.
The purpose of this study was to assess the effectiveness of two different relaxation techniques in the management of postoperative pain. The sample consisted of 40 women between the ages of 21 and 65 years who were undergoing elective cholecystectomy. Using an experimental design, participants were randomly assigned to four groups: an experimental group who received a taped recording of a rhythmic breathing exercise (RB); an experimental group who received a taped recording of Benson's Relaxation Technique (BRT); an attention-distraction control group (CA) who received a taped recording of a history of the hospital; and a standard control (CB) group who had only the routine perioperative care which all groups received. Data were collected on postoperative sensation and distress at five time points during the first 72 postoperative hours, number of doses of analgesic medication during the same time period, and number of postoperative hospital days. Data were analysed using multivariate and univariate analyses of variance. The BRT group was significantly different from the CA group on a combined sensation and distress factor (P = 0.011). No significant differences were found among groups for doses of analgesics (P = 0.068) or postoperative hospital days (P = 0.56).


Inhibitory Effects of Tofogliflozin on Cardiac Hypertrophy in Dahl Salt-Sensitive and Salt-Resistant Rats Fed a High-Fat Diet.
Sodium-glucose cotransporter 2 (SGLT2) inhibitors are drugs for diabetes and might prevent heart failure. In this study, we investigated the effects of tofogliflozin, an SGLT2 inhibitor, on cardiac hypertrophy and metabolism in hypertensive rats fed a high-fat diet. Dahl salt-sensitive (DS) rats, hypertensive model rats, and Dahl salt-resistant (DR) rats, non-hypertensive model rats, were fed a high-salt and high-fat diet containing tofogliflozin (0.005%) for 9 weeks to examine the effects of this drug on cardiac hypertrophy and metabolism. Tofogliflozin tended to suppress a rise of the systolic blood pressure, relative to the control, throughout the treatment period in both DR and DS rats, and significantly suppress a rise of the systolic blood pressure, relative to the control, at the 9th week in DS rats. Tofogliflozin reduced cardiac hypertrophy (heart weight/body weight) not only in DS rats but also in DR rats. Histological analysis showed that tofogliflozin significantly decreased cardiomyocyte hypertrophy and perivascular fibrosis in both DS and DR rats. Tofogliflozin significantly decreased the expression levels of genes related to cardiac hypertrophy (encoding for natriuretic peptides A and B and interleukin-6), and to cardiac fibrosis (encoding for transforming growth factor-β1 and collagen type IV), in DS rats. Recent studies have shown that hypertrophied and failing hearts shift to oxidizing ketone bodies as a significant fuel source. We also performed metabolome analysis for ventricular myocardial tissue. Tofogliflozin reduced 3-hydroxybutyrate, a ketone body, and significantly decreased the expression levels of β-hydroxybutyrate dehydrogenase 1 and 3-oxoacid CoA-transferase, which are related to ketone oxidization. In conclusion, tofogliflozin ameliorated cardiac hypertrophy and fibrosis along with reduction of ketone usage in myocardial tissue.


Molecular evidence that Reticulomyxa filosa is a freshwater naked foraminifer.
Reticulomyxa filosa is a freshwater protist possessing fine granular, branching and anastomosing pseudopodia and therefore traditionally placed in the class Granuloreticulosea, order Athalamida, as a sister group to the order Foraminiferida. Recent studies have revealed remarkable similarities in pseudopodial motility and ultrastructure between R. filosa and foraminifera (e.g. Allogromia laticollaris), prompting us to conduct a molecular phylogenetic analysis of these seemingly disparate organisms. We sequenced the complete small-subunit of the ribosomal DNA of the cultured strain of R. filosa and compared it to the corresponding sequences of other protists including 12 species of foraminifera. We also sequenced and analyzed the actin coding genes from R. filosa and two species of foraminifera, Allogromia sp. and Ammonia sp. The analysis of both data sets clearly shows that R. filosa branches within the clade of foraminifera, suggesting that R. filosa is in fact a freshwater naked foraminiferan.


Evolution of juvenile growth rates in female guppies (Poecilia reticulata): predator regime or resource level?
Recent theoretical and empirical work argues that growth rate can evolve and be optimized, rather than always being maximized. Chronically low resource availability is predicted to favour the evolution of slow growth, whereas attaining a size-refuge from mortality risk is predicted to favour the evolution of rapid growth. Guppies (Poecilia reticulata) evolve differences in behaviour, morphology and life-history traits in response to predation, thus demonstrating that predators are potent agents of selection. Predators in low-predation environments prey preferentially on small guppies, but those in high-predation environments appear to be non-selective. Because guppies can outgrow their main predator in low- but not high-predation localities, we predict that predation will select for higher growth rates in the low-predation environments.However, low-predation localities also tend to have lower productivity than high-predation localities, yield-ing the prediction that guppies from these sites should have slower growth rates. Here we compare the growth rates of the second laboratory-born generation of guppies from paired high- and low-predation localities from four different drainages. In two out of four comparisons, guppies from high-predation sites grew significantly faster than their low-predation counterparts. We also compare laboratory born descendants from a field introduction experiment and show that guppies introduced to a low-predation environment evolved slower growth rates after 13 years, although this was evident only at the high food level. The weight of the evidence suggests that resource availability plays a more important role than predation in shaping the evolution of growth rates.


Focal areas of increased lipid concentration on the coating of microbubbles during short tone-burst ultrasound insonification.
Acoustic behavior of lipid-coated microbubbles has been widely studied, which has led to several numerical microbubble dynamics models that incorporate lipid coating behavior, such as buckling and rupture. In this study we investigated the relationship between microbubble acoustic and lipid coating behavior on a nanosecond scale by using fluorescently labeled lipids. It is hypothesized that a local increased concentration of lipids, appearing as a focal area of increased fluorescence intensity (hot spot) in the fluorescence image, is related to buckling and folding of the lipid layer thereby highly influencing the microbubble acoustic behavior. To test this hypothesis, the lipid microbubble coating was fluorescently labeled. The vibration of the microbubble (n = 177; 2.3-10.3 μm in diameter) upon insonification at an ultrasound frequency of 0.5 or 1 MHz at 25 or 50 kPa acoustic pressure was recorded with the UPMC Cam, an ultra-high-speed fluorescence camera, operated at ~4-5 million frames per second. During short tone-burst excitation, hot spots on the microbubble coating occurred at relative vibration amplitudes > 0.3 irrespective of frequency and acoustic pressure. Around resonance, the majority of the microbubbles formed hot spots. When the microbubble also deflated acoustically, hot spot formation was likely irreversible. Although compression-only behavior (defined as substantially more microbubble compression than expansion) and subharmonic responses were observed in those microbubbles that formed hot spots, both phenomena were also found in microbubbles that did not form hot spots during insonification. In conclusion, this study reveals hot spot formation of the lipid monolayer in the microbubble's compression phase. However, our experimental results show that there is no direct relationship between hot spot formation of the lipid coating and microbubble acoustic behaviors such as compression-only and the generation of a subharmonic response. Hence, our hypothesis that hot spots are related to acoustic buckling could not be verified.


Antitussive drugs--past, present, and future.
Cough remains a serious unmet clinical problem, both as a symptom of a range of other conditions such as asthma, chronic obstructive pulmonary disease, gastroesophageal reflux, and as a problem in its own right in patients with chronic cough of unknown origin. This article reviews our current understanding of the pathogenesis of cough and the hypertussive state characterizing a number of diseases as well as reviewing the evidence for the different classes of antitussive drug currently in clinical use. For completeness, the review also discusses a number of major drug classes often clinically used to treat cough but that are not generally classified as antitussive drugs. We also reviewed a number of drug classes in various stages of development as antitussive drugs. Perhaps surprising for drugs used to treat such a common symptom, there is a paucity of well-controlled clinical studies documenting evidence for the use of many of the drug classes in use today, particularly those available over the counter. Nonetheless, there has been a considerable increase in our understanding of the cough reflex over the last decade that has led to a number of promising new targets for antitussive drugs being identified and thus giving some hope of new drugs being available in the not too distant future for the treatment of this often debilitating symptom.


The Effect of the 2009 Influenza Pandemic on Absence from Work.
In July 2009, the World Health Organization declared the first flu pandemic in nearly 40 years. Although the health effects of the pandemic have been studied, there is little research examining the labor productivity consequences. Using unique sick leave data from the Chilean private health insurance system, we estimate the effect of the pandemic on missed days of work. We estimate that the pandemic increased mean flu days missed by 0.042 days per person-month during the 2009 peak winter months (June and July), representing an 800% increase in missed days relative to the sample mean. Calculations using the estimated effect imply a minimum 0.2% reduction in Chile's labor supply. Copyright © 2017 John Wiley & Sons, Ltd.


Cancer-Associated Mutations in Endometriosis without Cancer.
Endometriosis, defined as the presence of ectopic endometrial stroma and epithelium, affects approximately 10% of reproductive-age women and can cause pelvic pain and infertility. Endometriotic lesions are considered to be benign inflammatory lesions but have cancerlike features such as local invasion and resistance to apoptosis. We analyzed deeply infiltrating endometriotic lesions from 27 patients by means of exomewide sequencing (24 patients) or cancer-driver targeted sequencing (3 patients). Mutations were validated with the use of digital genomic methods in microdissected epithelium and stroma. Epithelial and stromal components of lesions from an additional 12 patients were analyzed by means of a droplet digital polymerase-chain-reaction (PCR) assay for recurrent activating KRAS mutations. Exome sequencing revealed somatic mutations in 19 of 24 patients (79%). Five patients harbored known cancer driver mutations in ARID1A, PIK3CA, KRAS, or PPP2R1A, which were validated by Safe-Sequencing System or immunohistochemical analysis. The likelihood of driver genes being affected at this rate in the absence of selection was estimated at P=0.001 (binomial test). Targeted sequencing and a droplet digital PCR assay identified KRAS mutations in 2 of 3 patients and 3 of 12 patients, respectively, with mutations in the epithelium but not the stroma. One patient harbored two different KRAS mutations, c.35G→T and c.35G→C, and another carried identical KRAS c.35G→A mutations in three distinct lesions. We found that lesions in deep infiltrating endometriosis, which are associated with virtually no risk of malignant transformation, harbor somatic cancer driver mutations. Ten of 39 deep infiltrating lesions (26%) carried driver mutations; all the tested somatic mutations appeared to be confined to the epithelial compartment of endometriotic lesions.


Specificity and enzymatic mechanism of the editing exonuclease of Escherichia coli DNA polymerase III.
Exonucleolytic editing is a major contributor to the fidelity of DNA replication by the multisubunit DNA polymerase (pol) III holoenzyme. To investigate the source of editing specificity, we have studied the isolated exonuclease subunit, epsilon, and the pol III core subassembly, which carries the epsilon, theta, and alpha (polymerase) subunits. Using oligonucleotides with specific terminal mismatches, we have found that both epsilon and pol III core preferentially excise a mispaired 3' terminus and therefore have intrinsic editing specificity. For both epsilon and pol III core, exonuclease activity is much more effective with single-strand DNA; with a double-strand DNA, the exonuclease is strongly temperature-dependent. We conclude that the epsilon subunit of pol III holoenzyme is itself a specific editing exonuclease and that the source of specificity is the greater melting capacity of a mispaired 3' terminus.


A complex phosphodiesterase system controls beta-adrenoceptor signalling in cardiomyocytes.
beta-Adrenergic signalling mediates the positive inotropic effect of catecholamines on cardiomyocytes, mainly through cAMP generation and subsequent activation of PKA (protein kinase A). Given the large diversity of PKA targets within cardiac cells, a precisely regulated and confined activity of such signalling pathways is essential for the specificity of response. PDEs (phosphodiesterases) constitute the only cAMP-degrading mechanism and are expressed in the cardiomyocyte in at least 5 family variants. Each PDE family is characterized by unique functional properties and contributes to a cAMP-degrading system enabling the modulation of PKA activity in a stimulus-dependent fashion.


[Control of HCV, HBV and HIV Infections in Hemodialysis].
Infections with blood-borne pathogens are still common among patients on maintenance dialysis all over the world. The control of infection due to blood-borne viruses (particularly HBV) within dialysis units has been a major goal in the management of patients with chronic kidney disease in the industrialized world. Standard precautions and specific procedures have been recommended to prevent infections with HBV, HCV and HIV within dialysis units. Isolation of HBsAg positive patients by dialysis rooms, staff and machines continues to be an important step to control HBV infection within dialysis units, according to the CDC and other regulatory agencies. Some prospective observational studies have reported the complete prevention of HCV transmission to hemodialysis patients in the absence of any isolation policy, and the use of dedicated dialysis machines for HCV-infected patients is not recommended by clinical guidelines. Isolation of HCV-infected patients should be considered in special circumstances only. Vaccination is an important tool against transmission of HBV among patients on long-term dialysis even if the immune response towards the hepatitis B vaccine remains unsatisfactory. Hemodialysis is considered a low risk setting for the transmission of human immunodeficiency virus (HIV) infection, providing that standard and specific procedures are carefully observed. HIV-infected patients do not have to be isolated from other patients or dialyzed separately on dedicated machines.


Course, complications, and outcome of juvenile arthritis-related uveitis.
To describe the clinical features of uveitis in patients with juvenile idiopathic arthritis (JIA). Retrospective chart review of a subset of 1,081 consecutive JIA patients who were younger than 18 years of age and had uveitis, with a minimum of 1-year follow-up at a single center. One hundred forty-two patients (13.1%) developed uveitis after a mean follow-up of 6.3 years (range, 0.10-23.2). Uveitis types were chronic anterior (97/142, 68.3%), acute anterior (23/142, 16.2%), recurrent anterior (17/142, 12%), and panuveitis (5/142, 3.5%). Uveitic complications were observed in 37.3% of cases (53/142) and 32.5% of eyes (74/228). When we compared uveitic eyes with complications to uveitic eyes without complications, we found the following significant differences: time interval from diagnosis of JIA to diagnosis of uveitis was shorter (mean, 1.3 years vs. 2.2 years; p = 0.003) and use of oral prednisone was greater (59.1% vs 15.6%; p < 0.0001) in the eyes with complications. Twenty-one children (21/142, 14.8%) with uveitis underwent a total of 62 ocular surgeries. Good visual acuity (20/40 or better) was found in 90.8% of eyes (159/175) and in both eyes of 87% of cases (94/108), impaired visual acuity in 6 eyes of 4 cases (6/175, 3.4%), and blindness in 10 eyes of 10 cases (10/175, 5.7%). Only 2 patients had reduced visual acuity in both eyes. Surgery was the single most important risk factor for reduced visual acuity at the last follow-up (p = 0.0086). Most uveitic eyes with JIA achieved good visual outcome despite uveitic complications.


Interventions to improve patient safety in transitional care--a review of the evidence.
When a patient's transition from the hospital to home is less than optimal, the repercussions can be far-reaching - hospital readmission, adverse medical events, and even mortality. Elderly, especially frail older patients with complex health care problems appear to be a group particularly at risk for adverse events in general, and during transitions across health providers in particular. We undertook a systematic review to identify interventions designed to improve patient safety during transitional care of the elderly, with a particular focus on discharge interventions. We searched the literature for qualitative and quantitative studies on the subject published over the past ten years. The review revealed a set of potential intervention types aimed at the improvement of communication that contribute to safe transitional care. Intervention types included profession-oriented interventions (e.g. education and training), organisational/culture interventions (e.g. transfer nurse, discharge protocol, discharge planning, medication reconciliation, standardized discharge letter, electronic tools), or patient and next of kin oriented interventions (e.g. patient awareness and empowerment, discharge support). Results strongly indicate that elderly discharged from hospital to the community will benefit from targeted interventions aimed to improve transfer across healthcare settings. Future interventions should take into account multi-component and multi-disciplinary interventions incorporating several single interventions combined.


Changing patterns in the practice of carotid endarterectomy in a large metropolitan area.
Changes in the practice of carotid endarterectomy were studied by review of all endarterectomies performed in the greater Cincinnati area during 1980 and from July 1983 through June 1984. The number of operations rose from 431 to 750 (74% increase). The perioperative stroke rate fell from 8.6% in 1980 to 5.1% in 1983-1984; operative mortality declined from 2.8% to 2.3%; and the combined stroke or death rate declined from 9.5% to 6.5%. Asymptomatic carotid artery disease was the indication for 50% of the endarterectomies during both time periods. The combined stroke or death rate for asymptomatic patients declined from 6.9% to 5.3%, but both rates were higher than the 3% suggested as acceptable for prophylactic carotid endarterectomy. We conclude that carotid endarterectomy is becoming an increasingly common procedure, that morbidity continues to decline, and that mortality continues to be significant. Citywide surgical morbidity and mortality remain excessive for patients with asymptomatic carotid disease.


Extracting the structure of highly reflecting fiber Bragg gratings by measuring both the transmission and the reflection spectra.
We demonstrate a novel method that enables one to measure the structure of highly reflecting fiber Bragg gratings. The method is based on measuring both the transmission and reflection spectra of the grating and applying an inverse-scattering algorithm. The use of the transmission spectrum significantly reduces the sensitivity of the reconstruction to measurement noise, and therefore it significantly decreases the measurement duration. We experimentally demonstrate our method for reconstructing the structure of an apodized grating with a reflectivity of 99.91%.


A cluster randomised controlled trial of two rounds of mass drug administration in Zanzibar, a malaria pre-elimination setting-high coverage and safety, but no significant impact on transmission.
Mass drug administration (MDA) has the potential to interrupt malaria transmission and has been suggested as a tool for malaria elimination in low-endemic settings. This study aimed to determine the effectiveness and safety of two rounds of MDA in Zanzibar, a pre-elimination setting. A cluster randomised controlled trial was conducted in 16 areas considered as malaria hotspots, with an annual parasite index of > 0.8%. The areas were randomised to eight intervention and eight control clusters. The intervention included two rounds of MDA with dihydroartemisinin-piperaquine and single low-dose primaquine 4 weeks apart in May-June 2016. Primary and secondary outcomes were cumulative confirmed malaria case incidences 6 months post-MDA and parasite prevalences determined by PCR 3 months post-MDA. Additional outcomes included intervention coverage, treatment adherence, occurrence of adverse events, and cumulative incidences 3, 12, and 16 months post-MDA. Intervention coverage was 91.0% (9959/10944) and 87.7% (9355/10666) in the first and second rounds, respectively; self-reported adherence was 82.0% (881/1136) and 93.7% (985/1196). Adverse events were reported in 11.6% (147/1268) and 3.2% (37/1143) of post-MDA survey respondents after both rounds respectively. No serious adverse event was reported. No difference in cumulative malaria case incidence was observed between the control and intervention arms 6 months post-MDA (4.2 and 3.9 per 1000 population; p = 0.94). Neither was there a difference in PCR-determined parasite prevalences 3 months post-MDA (1.4% and 1.7%; OR = 1.0, p = 0.94), although having received at least the first MDA was associated with reduced odds of malaria infection (aOR = 0.35; p = 0.02). Among confirmed malaria cases at health facilities, 26.0% and 26.3% reported recent travel outside Zanzibar in the intervention and control shehias (aOR ≥ 85; p ≤ 0.001). MDA was implemented with high coverage, adherence, and tolerability. Despite this, no significant impact on transmission was observed. The findings suggest that two rounds of MDA in a single year may not be sufficient for a sustained impact on transmission in a pre-elimination setting, especially when the MDA impact is restricted by imported malaria. Importantly, this study adds to the limited evidence for the use of MDA in low transmission settings in sub-Saharan Africa. ClinicalTrials.gov, NCT02721186 (registration date: March 29, 2016).


Antineoplaston induces G(1) arrest by PKCalpha and MAPK pathway in SKBR-3 breast cancer cells.
Antineoplastons such as A10 include naturally occurring peptides and amino acid derivatives that control the neoplastic growth of cells. The mechanism underlying this antitumor effect was investigated using the breast cancer cell line, SKRB-3. Cells treated with A10 were monitored for any changes in cell cycle, expression of protein kinase C (PKC), or intracellular signal transduction, particularly phos-phorylation of mitogen-activated protein kinase (MAPK). The A10 markedly inhibited SKBR-3 proliferation due to arrest in the G(1) phase. A10 down-regulated the expression of PKCalpha protein, resulting in inhibition of extracellular signal-regulated kinase (ERK) MAPK phosphorylation. This increased the expression of p16 and p21 protein, with resultant inhibition of Rb phosphorylation, leading to G(1) arrest. This study has defined a pathway in which A10 arrested SKBR-3 cells in the G(1) phase via PKCalpha and MAPK. Our findings indicate that the antineoplaston A10 antitumor effect could be utilized as an effective therapy for breast cancer patients.


Germinal transpositions of the maize element Dissociation from T-DNA loci in tomato.
We have analyzed the pattern of germinal transpositions of artificial Dissociation (Ds) transposons in tomato. T-DNA constructs carrying Ds were transformed into tomato, and the elements were trans-activated by crossing to lines transformed with a stabilized Activator (sAc) that expressed the transposase gene. The sAc T-DNA carried a GUS gene to monitor its segregation. The Ds elements were inserted in a marker gene so that excision from the T-DNA could be monitored. The Ds elements also carried a genetic marker that was intended to be used for reinsertion selection of the elements after excision. Unfortunately, this gene was irreversibly inactivated on crossing to sAc. Germinal excision frequencies of Ds averaged 15-40%, but there was large variation between and within plants. Southern hybridization analysis of stable transposed Ds elements indicated that although unique transpositions predominate, early transposition events can lead to large clonal sectors in the germline of developing plants and to sibling offspring carrying the same transposition event. Multiple germinal transpositions from three different loci were examined for uniqueness, and 15 different transpositions were identified from each of three T-DNA loci that carried a single independent Ds. These were mapped relative to the donor T-DNA loci, and for each locus 70-80% of the transposed elements were closely linked to the donor site.


Epidermal growth factor activates phospholipase C in rat hepatocytes via a different mechanism from that in A431 or rat1hER cells.
Epidermal growth factor (EGF) can stimulate inositol lipid hydrolysis in rat hepatocytes and can accelerate GTP/GDP exchange in hepatic membranes. Both of these responses can be abolished by pretreatment with pertussis toxin, suggesting that EGF may regulate phospholipase C (PLC) activity via a guanine nucleotide-binding regulatory protein (G protein) in liver cells. In contrast, in A431 human epidermoid carcinoma cells EGF can induce a rapid phosphorylation of PLC-gamma on tyrosine residues that increases the activity of immunoprecipitated PLC-gamma, suggesting that tyrosine phosphorylation of PLC-gamma may be the mechanism for EGF-stimulated inositol trisphosphate production in these cells. To determine the importance of the phosphorylation of PLC-gamma on tyrosine residues in a system where the EGF receptor apparently couples to a G protein, the effect of EGF on tyrosine phosphorylation of PLC-gamma was examined in rat hepatocytes. PLC-gamma was immunoprecipitated from cell lysates with a PLC-gamma antiserum and its tyrosine phosphorylation state was determined using both Western blot analysis with phosphotyrosine antibodies and direct measurement of phosphorylated amino acids. The results were compared with analogous experiments performed with A431 cells and another cultured cell line expressing high levels of human EGF receptors, Rat1hER fibroblasts. Although the amount of PLC-gamma in rat hepatocytes is similar to that in A431 cells and slightly higher than that in Rat1hER cells, EGF causes a barely detectable increase in the phosphorylation of PLC-gamma on tyrosine in hepatocytes, whereas it stimulates a significant degree of phosphorylation of PLC-gamma on tyrosine in Rat1hER or A431 cells. Pretreatment of hepatocytes with pertussis toxin abolishes the ability of EGF to activate PLC, as determined by an increase in intracellular Ca2+, but has no effect on the small amount of phosphate incorporated into tyrosine residues on the PLC-gamma protein, demonstrating that this low level of PLC-gamma phosphorylation does not correlate with changes in PLC activity. The data suggest that phosphorylation of PLC-gamma on tyrosine is not important for EGF-enhanced PLC activity in hepatocytes. This conclusion implies that EGF may use a mechanism to regulate PLC activity in hepatocytes that is different from that used in cultured cells expressing high levels of EGF receptors.


[Obstructive sleep apnea syndrome in young infants].
Obstructive sleep apnea syndrome (OSAS) has been reported among children aged 3-6 years. Adenotonsillar hypertrophy is the most common cause of OSAS. The upper airway obstruction results in repeated obstructive apneas and hypopneas, associated with oxygen desaturations and sleep fragmentation. OSAS in children is associated with higher respiratory morbidity as well as significant clinical consequences, mainly neurocognitive and behavioral problems, impaired growth and cardiac dysfunction. The objectives of the present study were to determine the clinical and sleep characteristics of OSAS in children younger than 2 years, and to evaluate morbidity and health care utilization of infants with OSAS. This retrospective study included 35 children younger than 2 years of age, referred for polysomnography because of suspected OSAS. The controL group included healthy children, matched by age, gender and pediatrician. A total of 33 infants were diagnosed with OSAS. Mean apnea/hypopnea index [AHI) was 18.7 +/- 18.1 events/ hour [range 1.3-90.2]. In 10 infants a pattern of intermittent hypoxemia was observed. Infants with OSAS demonstrated a higher number of primary care clinic visits (20.8 +/- 14 vs. 12.1 +/- 6.6, P < 0.02). A higher percentage of children with OSAS visited the emergency room (60.6% vs. 32.2%, P < 0.03), and was hospitalized (36.3% vs. 12.9%, P < 0.03). Duration of hospitalization was also higher in the OSAS group [23.2 +/- 14.1% vs. 3 +/- 2.1%, P < 0.05). The number of drug prescriptions was higher among the study group [25.9 +/- 21.8% vs. 13.6 +/- 10.1, P < 0.03). Thirteen patients underwent adenoidectomy with or without tonsillectomy, resulting in improvement of AHI, decreasing from 26.4 +/- 24 before to 3.6 +/- 4.5 events per hour after surgery (p < 0.01). OSAS can be found in infants younger than 2 years of age. OSAS at this young age is characterized by a higher morbidity in comparison to healthy children. Early diagnosis and treatment may prevent morbidity in young children with OSAS.


Ni2+-Dependent and PsaR-Mediated Regulation of the Virulence Genes pcpA, psaBCA, and prtA in Streptococcus pneumoniae.
Previous studies have shown that the transcriptional regulator PsaR regulates the expression of the PsaR regulon consisting of genes encoding choline binding protein (PcpA), the extracellular serine protease (PrtA), and the Mn2+-uptake system (PsaBCA), in the presence of manganese (Mn2+), zinc (Zn2+), and cobalt (Co2+). In this study, we explore the Ni2+-dependent regulation of the PsaR regulon. We have demonstrated by qRT-PCR analysis, metal accumulation assays, β-galactosidase assays, and electrophoretic mobility shift assays that an elevated concentration of Ni2+ leads to strong induction of the PsaR regulon. Our ICP-MS data show that the Ni2+-dependent expression of the PsaR regulon is directly linked to high, cell-associated, concentration of Ni2+, which reduces the cell-associated concentration of Mn2+. In vitro studies with the purified PsaR protein showed that Ni2+ diminishes the Mn2+-dependent interaction of PsaR to the promoter regions of its target genes, confirming an opposite effect of Mn2+ and Ni2+ in the regulation of the PsaR regulon. Additionally, the Ni2+-dependent role of PsaR in the regulation of the PsaR regulon was studied by transcriptome analysis.


Initiation of drug dealing among a prospective cohort of street-involved youth.
Street-involved youth who use drugs may have limited income-generation options and are known to commonly become immersed in illicit drug markets to generate funds. However, little attention has been given to factors that may drive drug dealing initiation among this vulnerable population. This longitudinal study examines drug dealing initiation among street-involved youth. Data were derived from the At-Risk Youth Study from September 2005 to November 2014; a prospective cohort of 194 street-involved youth who use drugs aged 14-26, in Vancouver, Canada. Extended Cox model was used to identify factors independently associated with time to first drug dealing. Among street-involved youth who had never dealt drugs at baseline, 56 (29%) individuals initiated drug dealing during the study period for an incidence density of 13.0 per 100 person-years (95% confidence interval [CI]: 9.9-17.2). In multivariable Cox regression analysis, male gender (adjusted hazard ratio [AHR] = 1.90, 95% CI: 1.06-3.42), homelessness (AHR = 1.88, 95% CI: 1.05-3.35), crystal methamphetamine use (AHR = 2.48, 95% CI: 1.47-4.20), and crack cocaine use (AHR = 2.35, 95% CI: 1.38-4.00) were positively and independently associated with initiating drug dealing. Homelessness and stimulant drug use were key risk factors for drug dealing initiation among street-involved youth. Findings indicate that evidence-based and innovative interventions, including youth-centric supportive housing, low threshold employment programs, and stimulant addiction treatment should be implemented and evaluated as strategies to help prevent this vulnerable population from engaging in risky illegal income generation practices.


MHC class II antigen presentation by intestinal epithelial cells fine-tunes bacteria-reactive CD4 T cell responses.
Although intestinal epithelial cells (IECs) can express major histocompatibility complex class II (MHC II), especially during intestinal inflammation, it remains unclear if antigen presentation by IECs favours pro- or anti-inflammatory CD4+ T cell responses. Using selective gene ablation of MHC II in IECs and IEC organoid cultures, we assessed the impact of MHC II expression by IECs on CD4+ T cell responses and disease outcomes in response to enteric bacterial pathogens. We found that intestinal bacterial infections elicit inflammatory cues that greatly increase expression of MHC II processing and presentation molecules in colonic IECs. Whilst IEC MHC II expression had little impact on disease severity following Citrobacter rodentium or Helicobacter hepaticus infection, using a colonic IEC organoid-CD4+ T cell co-culture system, we demonstrate that IECs can activate antigen-specific CD4+ T cells in an MHC II-dependent manner, modulating both regulatory and effector Th cell subsets. Furthermore, we assessed adoptively transferred H. hepaticus-specific CD4+ T cells during intestinal inflammation in vivo and report that IEC MHC II expression dampens pro-inflammatory effector Th cells. Our findings indicate that IECs can function as non-conventional antigen presenting cells and that IEC MHC II expression fine-tunes local effector CD4+ T cell responses during intestinal inflammation.


Motor cortex stimulation for trigeminal neuropathic or deafferentation pain: an institutional case series experience.
Trigeminal neuropathy is a rare, devastating condition that can be intractable and resistant to treatment. When medical treatment fails, invasive options are limited. Motor cortex stimulation (MCS) is a relatively recent technique introduced to treat central neuropathic pain. The use of MCS to treat trigeminal neuropathic or deafferentation pain is not widespread and clinical data in the medical literature that demonstrate efficacy are limited. We retrospectively reviewed patients with trigeminal neuropathic or trigeminal deafferentation pain who were treated at the Oregon Health & Science University between 2001 and 2008 by 1 neurosurgeon using MCS. Eight of 11 patients (3 male, 8 female) underwent successful permanent implantation of an MCS system. All 8 patients reported initial satisfactory pain control. Three failed to experience continued pain control (6 months of follow-up). Five continued to experience long-term pain control (mean follow-up, 33 months). Average programming sessions were 2.2/year (all 8 patients) and 1.55/year (5 patients who sustained long-term pain control). Patients with anesthesia dolorosa or trigeminal deafferentation pain who had previously undergone ablative trigeminal procedures responded poorly to MCS. We encountered no perioperative complications. MCS is a safe and potentially effective therapy in certain patients with trigeminal neuropathy.


Hemodynamic study of TCPC using in vivo and in vitro 4D Flow MRI and numerical simulation.
Altered total cavopulmonary connection (TCPC) hemodynamics can cause long-term complications. Patient-specific anatomy hinders generalized solutions. 4D Flow MRI allows in vivo assessment, but not predictions under varying conditions and surgical approaches. Computational fluid dynamics (CFD) improves understanding and explores varying physiological conditions. This study investigated a combination of 4D Flow MRI and CFD to assess TCPC hemodynamics, accompanied with in vitro measurements as CFD validation. 4D Flow MRI was performed in extracardiac and atriopulmonary TCPC subjects. Data was processed for visualization and quantification of velocity and flow. Three-dimensional (3D) geometries were generated from angiography scans and used for CFD and a physical model construction through additive manufacturing. These models were connected to a perfusion system, circulating water through the vena cavae and exiting through the pulmonary arteries at two flow rates. Models underwent 4D Flow MRI and image processing. CFD simulated the in vitro system, applying two different inlet conditions from in vitro 4D Flow MRI measurements; no-slip was implemented at rigid walls. Velocity and flow were obtained and analyzed. The three approaches showed similar velocities, increasing proportionally with high inflow. Atriopulmonary TCPC presented higher vorticity compared to extracardiac at both inflow rates. Increased inflow balanced flow distribution in both TCPC cases. Atriopulmonary IVC flow participated in atrium recirculation, contributing to RPA outflow; at baseline, IVC flow preferentially traveled through the LPA. The combination of patient-specific in vitro and CFD allows hemodynamic parameter control, impossible in vivo. Physical models serve as CFD verification and fine-tuning tools.


Parasite resistance and the adaptive significance of sleep.
Sleep is a biological enigma. Despite occupying much of an animal's life, and having been scrutinized by numerous experimental studies, there is still no consensus on its function. Similarly, no hypothesis has yet explained why species have evolved such marked variation in their sleep requirements (from 3 to 20 hours a day in mammals). One intriguing but untested idea is that sleep has evolved by playing an important role in protecting animals from parasitic infection. This theory stems, in part, from clinical observations of intimate physiological links between sleep and the immune system. Here, we test this hypothesis by conducting comparative analyses of mammalian sleep, immune system parameters, and parasitism. We found that evolutionary increases in mammalian sleep durations are strongly associated with an enhancement of immune defences as measured by the number of immune cells circulating in peripheral blood. This appeared to be a generalized relationship that could be independently detected in 4 of the 5 immune cell types and in both of the main sleep phases. Importantly, no comparable relationships occur in related physiological systems that do not serve an immune function. Consistent with an influence of sleep on immune investment, mammalian species that sleep for longer periods also had substantially reduced levels of parasitic infection. These relationships suggest that parasite resistance has played an important role in the evolution of mammalian sleep. Species that have evolved longer sleep durations appear to be able to increase investment in their immune systems and be better protected from parasites. These results are neither predicted nor explained by conventional theories of sleep evolution, and suggest that sleep has a much wider role in disease resistance than is currently appreciated.


Cystatin C is associated with risk of venous thromboembolism in subjects with normal kidney function--the Tromsø study.
Previous studies have shown an association between impaired kidney function, assessed by cystatin C-based estimated glomerular filtration rate, and venous thromboembolism. The aim of this study was to investigate whether serum cystatin C was associated with a risk of venous thromboembolism among subjects with normal kidney function in a prospective population-based study. Cystatin C was measured in serum from 3251 men and women with normal kidney function, aged 25-84 years, who participated in the Tromsø study in 1994-1995. Normal kidney function was defined as a creatinine-based estimated glomerular filtration rate greater than 90 mL/min/1.73 m(2) and absence of microalbuminuria. Incident venous thromboembolism was registered from the date of inclusion through to the end of follow-up, September 1, 2007. Cox-regression models were used to calculate hazard ratios with 95% confidence intervals for venous thromboembolism. There were 83 incident venous thromboembolic events, of which 53 (63.9 %) were provoked, during a median of 12.3 years of follow-up. A one standard deviation (0.11 mg/L) increase in serum cystatin C levels was associated with a 43% (hazard ratio 1.43; 95% confidence interval 1.17-1.72) increased risk of total venous thromboembolism. Subjects with cystatin C levels in the top quartile (≥ 0.87 mg/L) had a 2.5-fold (hazard ratio 2.51; 95% confidence interval 1.27-4.96) increased risk of venous thromboembolism compared to those with levels in the bottom quartile (≤ 0.72 mg/L) in adjusted analysis. The risk estimates were even higher for provoked venous thromboembolism (hazard ratio 3.11; 95% confidence interval 1.23-7.86). Serum cystatin C levels were associated with the risk of venous thromboembolism in subjects with normal kidney function. Our findings suggest that elevated serum cystatin C levels may promote venous thrombosis beyond reflecting impaired kidney function.


FermiKit: assembly-based variant calling for Illumina resequencing data.
FermiKit is a variant calling pipeline for Illumina whole-genome germline data. It de novo assembles short reads and then maps the assembly against a reference genome to call SNPs, short insertions/deletions and structural variations. FermiKit takes about one day to assemble 30-fold human whole-genome data on a modern 16-core server with 85 GB RAM at the peak, and calls variants in half an hour to an accuracy comparable to the current practice. FermiKit assembly is a reduced representation of raw data while retaining most of the original information. https://github.com/lh3/fermikit firstname.lastname@example.com.