Role for the P4 amino acid residue in substrate utilization by the poliovirus 3CD proteinase.
Amino acid insertions or substitutions were introduced into the poliovirus P1 capsid precursor at locations proximal to the two known Q-G cleavage sites to examine the role of the P4 residue in substrate processing by proteinase 3CD. Analysis of the processing profile of P1 precursors containing four-amino-acid insertions into the carboxy terminus of VP3 or a single-amino-acid substitution at the P4 position of the VP3-VP1 cleavage site demonstrates that substitution of the alanine residue in the P4 position of the VP3-VP1 cleavage site significantly affects cleavage at that site by proteinase 3CD. A single-amino-acid substitution at the P4 position of the VP0-VP3 cleavage site, on the other hand, has only a slight effect on 3CD-mediated processing at this cleavage site. Finally, analysis of six amino acid insertion mutations containing Q-G amino acid pairs demonstrates that the in vitro and in vivo selection of a cleavage site from two adjacent Q-G amino acid pairs depends on the presence of an alanine in the P4 position of the cleaved site. Our data provide genetic and biochemical evidence that the alanine residue in the P4 position of the VP3-VP1 cleavage site is a required substrate determinant for the recognition and cleavage of that site by proteinase 3CD and suggest that the P4 alanine residue may be specifically recognized by proteinase 3CD.


An ultrastructural and radioautographic study of the chromocentric interphase nucleus in plant meristematic cells (Raphanus sativus).
In Raphanus sativus, the mitotic chromosomes are quite short and, on reaching the cell poles, soon undergo extensive unravelling. By late telophase and early interphase, only a few chromosome segments, believed to correspond to the centromeric regions, are still visible in the form of chromocentres closely associated with the nuclear envelope. Although interphase nuclei show little internal structural differentiation, high-resolution radioautography has permitted us to establish which of them have reached the early, mid and late S periods. In early S nuclei, only the nucleolus and the euchromatin which pervades the nuclear cavity become labelled. By the mid S-period, the diffuse chromatin and nucleolus incorporate less thymidine and DNA synthesis is initiated within the peripheral chromocentres. Subsequently, the radioautographic grains become restricted to the chromocentres. The finding that certain late S nuclei exhibit loosely organized chromocentres strongly suggests that these heterochromatic chromosome segments undergo important conformational modifications during DNA replication. Finally, the presence of radioautographic grains over the lacunar regions of the nucleolus in early and mid S nuclei demonstrates that intranucleolar DNA replicates during the earlier portion of the S-period.


Community-based health insurance in poor rural China: the distribution of net benefits.
The collapse of China's Cooperative Medical System (CMS) in 1978 resulted in the lack of an organized financing scheme for health care, adversely affecting rural farmers' access to health care, especially among the poor. The Chinese government recently announced a policy to re-establish some forms of community-based insurance (CBI). Many existing schemes involve low premiums but high co-payments. We hypothesized that such benefit design leads to unequal distribution of the "net benefits" (NB)--benefits net of payment--because even though low premiums are more affordable to poor farmers, high co-payments may have a significant deterrent effect on the poor in the use of services in CBI. To test this hypothesis empirically, we estimated the probability of farmers joining a re-established CBI using logistic regression, and the utilization of health care services for those who joined the scheme using the two-part model. Based on the estimations, we predicted the distribution of NB among those who joined the CBI and for the entire population in the community. Our data came from a household survey of 4160 members of 1173 households conducted in six villages in Fengshan Township, Guizhou Province, China. Three principal findings emerged from this study. First, income is an important factor influencing farmers' decision to join a CBI despite the premium representing a very small fraction of household income. Secondly, both income and health status influence enrollees' utilization of health services: richer/sicker participants obtain greater NB from the CBI than poorer/healthier members, meaning that the poorer/healthier participants subsidize the rich/sick. Thirdly, wealthy farmers benefit the most from the CBI with low premium and high co-payment features at every level of health status. In conclusion, policy recommendations related to the improvement of the benefit distribution of CBI schemes are made based on the results from this study.


GPR56 is highly expressed in neural stem cells but downregulated during differentiation.
The G-protein-coupled receptor 56 (GPR56) plays important roles in brain development and tumorigenesis. cDNA data suggest that GPR56 has potential to become a neural stem cell (NSC) or neural progenitor cell (NPC) marker. However, expression of GPR56 protein in human NSC/NPCs was not explored. Using specific antibodies and immunochemistry, we showed that GPR56 was highly expressed in nestin-positive NSC/NPCs in the ventricular/subventricular zone of human and mouse fetal brains, and in cultured neurospheres derived from both human and mouse fetal brains. Downregulation of GPR56 protein occurred earlier than that of nestin in differentiating neurosphere cultures. Loss of GPR56 protein was also evident in well-differentiated glial fibrillary acidic protein-positive astrocytes and betaIII-tubulin-positive neurons. Our data suggest that GPR56 can be used as an NSC/NPC marker within the neural cell lineage, especially in combination with nestin.


Estimation of Aromatic Secondary Organic Aerosol Using a Molecular Tracer-A Chemical Transport Model Assessment.
A modified community multiscale air quality model, which can simulate the regional distributions of 2,3-dihydroxy-4-oxopentanoic acid (DHOPA), a marker species for monoaromatic secondary organic aerosol (SOA), was applied to assess the applicability of using the DHOPA to aromatic SOA mass ratio (fSOA) from smog chamber experiments to estimate aromatic SOA during a three-week wintertime air quality campaign in urban Shanghai. The modeled daily DHOPA concentrations based on the chamber-derived mass yields agree well with the organic marker field measurements (R = 0.79; MFB = 0.152; and MFE = 0.440). Two-thirds of the DHOPA are from the oxidation of ARO1 (lumped less-reactive aromatic species; mostly toluene), with the rest from ARO2 (lumped more-reactive aromatic species; mostly xylenes). Modeled DHOPA is mainly in the particle phase under ambient organic aerosol (OA) loading but could exhibit significant gas-particle partitioning when a higher estimation of the DHOPA vapor pressure is used. The modeled fSOA shows a strong dependence on the OA loading when only semivolatile aromatic SOA components are included in the fSOA calculations. However, this OA dependence becomes weaker when non-volatile oligomers and dicarbonyl SOA products are considered. A constant fSOA value of ∼0.002 is determined when all aromatic SOA components are included, which is a factor of 2 smaller than the commonly applied chamber-based fSOA value of 0.004 for toluene. This model-derived fSOA value does not show much spatial variation and is not sensitive to alternative estimates of DHOPA vapor pressures and SOA yields, and thus provides an appropriate scaling factor to assess aromatic SOA from DHOPA measurements. This result helps refine the quantification of SOA attributable to monoaromatic hydrocarbons in urban environments and thereby facilitates the evaluation of control measures targeting these specific precursors.


"That word, cancer": breast care behavior of Hispanic women in new Mexico background and literature review.
Despite international efforts, national and ethnic disparities in utilization of breast cancer (BC) screenings prevail. In the United States, Hispanic women have one of the lowest BC screening rates. The purpose of our study was to examine how Hispanic women in New Mexico described their breast care behavior (BCB; BC screening practices, motivation to act, and breast care information behavior). Analysis of focus groups revealed five types of approaches to BCB. These findings have global implications for health care practitioners in directing attention toward the complexity of BC preventive behavior. Implications for other ethnic groups are discussed.


Prevention of coronary thrombosis with subthrombolytic doses of tissue-type plasminogen activator.
To determine whether tissue-type plasminogen activator (t-PA) may prevent coronary thrombosis or accelerate the lysis of clot formed under conditions in which increased concentration of the activator is present before thrombosis, clot lysis studies were undertaken in vitro and in vivo. In vitro, exogenous t-PA (6 to 100,000 ng/ml) accelerated the lysis of clot in a dose-dependent fashion when the clot was formed either from whole plasma or from euglobulin fractions (n = 316 determinations). Adding t-PA before clot formation shortened the time to lysis by at least threefold with euglobulin fractions and by at least 10-fold with whole plasma clots, which is consistent with the presence of inhibitors of fibrinolysis in whole plasma and with the binding of t-PA to nascent fibrin. In an intact dog preparation of coronary thrombosis (n = 25), occlusive thrombus formation was prevented when t-PA was present in subthrombolytic concentrations (430 to 1200 ng/ml, n = 5). Occlusive thrombus formation occurred after only discontinuation of the t-PA infusion and clearance of t-PA. Lower concentrations of t-PA (147 to 427 ng/ml, n = 6) significantly delayed occlusion (26 +/- 6.5 vs 7.8 +/- 2.8 min for controls). In animals with t-PA concentrations of less than 140 ng/ml (n = 4), the time to occlusion was unaltered (7.7 +/- 4.5 min). The present study demonstrates that t-PA present before clot formation inhibits thrombosis or accelerates thrombolysis depending on concentration, and that subthrombolytic doses of t-PA can prevent thrombus formation in vivo.(ABSTRACT TRUNCATED AT 250 WORDS)


[Long-Term Survival with Multidisciplinary Treatment and Nutritional Therapy for Thoracic Esophageal Cancer with Multiple Liver Metastases-A Case Report].
The patient was a 63-year-old man with a chief complaint. Upper endoscopic examination revealed a semicircular type 2 lesion, sized 24-28 cm, on the incisor teeth and a 3 cm sized elevated lesion directly above the EGJ. When biopsy was performed, squamous cell carcinoma(SCC)was detected. In this case, lymph node metastasis and multiple liver metastases were observed, and diagnosis at the first examination was cT3N2M1(HEP), Stage Ⅳ. After 7 months of chemotherapy, he underwent right thoracic esophageal subtotal resection, 3-field lymph node dissection, posterior mediastinal gastric tube reconstruction, and partial hepatectomy. Despite receiving postoperative chemotherapy, he showed recurrence in the liver(S8). Four additional courses of chemotherapy were administered and partial hepatectomy(S8)was performed, without the appearance of new lesions. He was considered to be cured 1 year and 6months after starting the treatment and was followed- up without chemotherapy. However, 4 months later, chemotherapy was resumed when right adrenal and abdominal wall metastases and liver recurrence(S3)were found. After that, the regimen was modified, and he continued treatment. More than 4 years have passed since the start of treatment, but the treatment has been continued without a decline in ADL.


Impact of pharmacy student and resident-led discharge counseling on heart failure patients.
Many health systems have implemented interventions to reduce the rate of heart failure readmissions. Pharmacists have the training and expertise to provide effective medication-related education. However, few studies have examined the impact of discharge education provided by pharmacy students and residents on patients hospitalized with heart failure exacerbations. This was a nonrandomized intervention study evaluating the impact of a pharmacy student and resident-led discharge counseling program on heart failure readmissions. The primary end point was the 30-day heart failure readmission rate. Secondary end points included self-reported patient understanding of medications, number of medication errors documented, and estimated associated cost avoidance. A total of 86 and 94 patients were enrolled into the intervention and control groups, respectively. No statistically significant difference in readmission rates was detected between the intervention and the control groups. Thirty-four medication errors and discrepancies were documented, or 1 for every 2.5 patients counseled, resulting in an estimated cost avoidance of $4241 for the institution. Eighty-nine percent of patients who received discharge counseling agreed they had a better understanding of their medications after speaking with a pharmacy resident or student. There was no statistically significant difference in readmission rates; however, several medication errors were prevented, and a large percentage of patients expressed an improved understanding of their medications.


Assembly of Protein-Polysaccharide Complexes for Delivery of Bioactive Ingredients: A Perspective Paper.
Protein-polysaccharide complexes can be created in various ways (physical mixing, enzymatic cross-linking, chemical cross-linking, and Maillard reaction), and diverse protein-polysaccharide complexes are generally grouped into non-covalent and covalent complexes. Delivery systems constructed through assembly of protein-polysaccharide complexes (DSAPC) consist of emulsion-based delivery systems, capsule-based delivery systems, molecular complexes, nanogels, core-shell particles, composite nanoparticles, and micelles. DSAPC are effective delivery vehicles in enhancing the overall efficacy of bioactive ingredients, and DSAPC may possess multiple advantages over other delivery vehicles in bioactive ingredient delivery. However, designing and applying DSAPC are still faced with some challenges, such as low loading of bioactive ingredients. Efforts are required to reconsider and improve efficiency of DSAPC in many aspects, such as controlled release and targeted delivery. On the basis of more comprehensive and deeper understandings, DSAPC can be designed more rationally for delivery of bioactive ingredients.


Development of a solid-phase microextraction LC-MS/MS method for determination of oxidative stress biomarkers in biofluids.
Recently the connection between oxidative stress and various diseases, including cancer and Alzheimer's, attracts notice as a pathway suitable for diagnostic purposes. 8-Oxo-deoxyguanosine and 8-oxo-deoxyadenosine produced from the interaction of reactive oxygen species with DNA become prominent as biomarkers. Several methods have been developed for their determination in biofluids, including solid-phase extraction and enzyme-linked immunosorbent assays. However, still, there is a need for reliable and fast analytical methods. In this context, solid-phase microextraction offers many advantages such as flexibility in geometry and applicable sample volume, as well as high adaptability to high-throughput sampling. In this study, a solid-phase microextraction method was developed for the determination of 8-oxo-deoxyguanosine and 8-oxo-deoxyadenosine in biofluids. The extractive phase of solid-phase microextraction consisted of hydrophilic-lipophilic balanced polymeric particles. In order to develop a solid-phase microextraction method suitable for the determination of the analytes in saliva and urine, several parameters, including desorption solvent, desorption time, sample pH, and ionic strength, were scrutinized. Analytical figures of merit indicated that the developed method provides reasonable interday and intraday precisions (<15% in both biofluids) with acceptable accuracy. The method provides a limit of quantification for both biomarkers at 5.0 and 10.0 ng/mL levels in saliva and urine matrices, respectively.


QTL for fatty acid composition and yield in linseed (Linum usitatissimum L.).
The combined SSR-SNP map and 20 QTL for agronomic and quality traits will assist in marker assisted breeding as well as map-based cloning of key genes in linseed. Flax is an important nutraceutical crop mostly because it is a rich source of omega-3 fatty acids and antioxidant compounds. Canada is the largest producer and exporter of oilseed flax (or linseed), creating a growing need to improve crop productivity and quality. In this study, a genetic map was constructed based on selected 329 single nucleotide polymorphic markers and 362 simple sequence repeat markers using a recombinant inbred line population of 243 individuals from a cross between the Canadian varieties CDC Bethune and Macbeth. The genetic map consisted of 15 linkage groups comprising 691 markers with an average marker density of one marker every 1.9 cM. A total of 20 quantitative trait loci (QTL) were identified corresponding to 14 traits. Three QTL each for oleic acid and stearic acid, two QTL each for linoleic acid and iodine value and one each for palmitic acid, linolenic acid, oil content, seed protein, cell wall, straw weight, thousand seed weight, seeds per boll, yield and days to maturity were identified. The QTL for cell wall, straw weight, seeds per boll, yield and days to maturity all co-located on linkage group 4. Analysis of the candidate gene regions underlying the QTL identified proteins involved in cell wall and fibre synthesis, fatty acid biosynthesis as well as their metabolism and yield component traits. This study provides the foundation for assisting in map-based cloning of the QTL and marker assisted selection of a wide range of quality and agronomic traits in linseed and potentially fibre flax.


Are lawyers enemies of psychiatrists? A survey of civil commitment counsel and judges.
The author surveyed 101 attorneys and judges involved in civil commitment procedures to answer the question of how they view psychiatrists, mental hospitals, and the mentally ill. She found that these lawyers and judges tended to view psychiatrists in favorable terms, that their attitudes toward mental hospitals reflected their perception of the weaknesses of these hospitals and their view of hospitalization as a last resort, and that they tended to reject negative statements about the mentally ill. She discusses the effect of these attitudes on the behavior of lawyers and judges toward psychiatrists and the mentally ill in civil commitment hearings.


Production of monocyte chemoattractant protein-1 in amyotrophic lateral sclerosis.
The presence of activated microglia in the spinal cord of amyotrophic lateral sclerosis (ALS) patients is usually accompanied by inflammatory biochemical changes, but these are largely unexplored. Monocyte chemoattractant protein-1 (MCP-1) is critical for recruitment of inflammatory cells of monocytic lineage after inflammation or injury to the central nervous system. MCP-1 concentrations were measured by an enzyme-linked immunosorbent assay in the cerebrospinal fluid (CSF) and the serum of 27 patients with ALS and 30 patients with noninflammatory neurological diseases. In ALS, circulating MCP-1 levels were significantly increased in the serum and particularly in the CSF. Immunoreactivity for MCP-1 in ALS spinal cord was detected mostly in astrocytes but also in microglia, neurons, and within the vasculature of the cord. Our findings suggest a role for MCP-1 as an important molecular mediator of the injury response in ALS.


Effect of melatonin supplementation on sleep quality: a systematic review and meta-analysis of randomized controlled trials.
The Present study was conducted to systematically review the effect of the melatonin on sleep quality. We summarized evidence from randomized clinical trials (RCTs) that investigated the effects of melatonin on sleep quality as assessed by the Pittsburgh Sleep Quality Index (PSQI) in adults with various diseases. The literature searches of English publications in MEDLINE and EMBASE databases were performed up June 2020. Results were summarized as mean differences (MD) with 95% confidence intervals (CI) using random effects model (DerSimonian-Laird method). Heterogeneity among studies was evaluated by the Cochrane Q test and I-squared (I2). To determine the predefined sources of heterogeneity, subgroup analysis was performed. Of 2642 papers, 23 RCTs met inclusion criteria. Our results indicated that melatonin had significant effect on sleep quality as assessed by the Pittsburgh Sleep Quality Index (PSQI) (WMD: - 1.24; 95% CI - 1.77, - 0.71, p = 0.000). There was significant heterogeneity between studies (I2 = 80.7%, p = 0.000). Subgroup analysis based on health status and kind of intervention were potential between-study heterogeneity. Subgroup analysis based on health status revealed melatonin intervention in subjects with Respiratory diseases (WMD: - 2.20; 95% CI - 2.97, - 1.44, p = 0.000), Metabolic disorders (WMD: - 2.74; 95% CI - 3.48, - 2.00, p = 0.000) and sleep disorders (WMD: - 0.67; 95% CI - 0.98, - 0.37, p = 0.000) has significant effect on sleep quality. We found that the treatment with exogenous melatonin has positive effects on sleep quality as assessed by the Pittsburgh Sleep Quality Index (PSQI) in adult. In adults with respiratory diseases, metabolic disorders, primary sleep disorders, not with mental disorders, neurodegenerative diseases and other diseases.


[Melanoma of ganglion cells arising from a thoracic sympathetic ganglion in an adult--a case report].
One case of pigmented tumor arising from the thoracic sympathetic ganglion in a 33 years old man is reported. By microscopy, the tumor cells were arranged in nests, among which ganglionic cells were observed. By Masson-Fontana and Lillie's stain, the pigment in the tumor cells was positive for melanin. By Glees-Marsland's stain, neurofibrils were demonstrated in the cytoplasm of some tumor cells. The presence of melanin and neurofibrils in the tumor cells and the interstitium proves that the tumor is a real melanocytic one arising from the sympathetic ganglion and that both the ganglionic and melanocytic cells originate from the neural crest. The malignancy of this tumor was also evidenced by the rapid extensive metastases occurring half a month after the operation.


The effects of caloric restriction and its mimetics in Alzheimer's disease through autophagy pathways.
Alzheimer's disease (AD) is a neurodegenerative disorder that commonly occurs among older individuals. Increasing evidence suggests that a low-caloric diet might be a promising adjuvant therapeutic strategy for slowing or preventing the pathogenesis and progression of AD through the induction of autophagy. Several intracellular pathways have been implicated in caloric restriction (CR)-induced autophagy. In this review, we summarized the efficacy of CR as well as its mimetics (resveratrol, spermidine, aspirin, rapamycin, metformin, and curcumin) in improving cognitive function of rodent models of AD. On the basis of recent in vitro and animal studies, the beneficial effects of CR- or caloric restriction mimetics-induced autophagy in alleviating amyloid burden and tau pathology of AD were also discussed.


Indigenous youth participatory action research: re-visioning social justice for social work with indigenous youths.
The NASW Code of Ethics identifies social justice as one of six foundational values of the social work profession. Indigenous communities have long questioned the authenticity of this commitment and rightly so, given the historical activities of social work and social workers. Still, the commitment persists as an inspiration for an imperfect, yet determined, profession. This article presents a theoretical discussion of questions pertinent for social justice in social work practice in Native American communities: Whose definition of social justice should prevail in work with and in Indigenous communities? What can a revisioning of social justice mean to the development of Native communities and for Native youths in particular? What methods or processes of social work are most appropriate for this social justice work? This article presents a case for the practice of youth participatory action research as one method to work for social justice in Native communities.


Molecular chaperones and co-chaperones in Parkinson disease.
Parkinson disease, a progressive neurodegenerative disorder, is caused by the pathological accumulation of proteins, including the ubiquitous presynaptic protein α-synuclein. Alterations in the metabolism of α-synuclein have clearly been linked to neurodegeneration, and early steps in the pathological sequence of this protein include the formation of oligomers, fibrils, and small aggregates. Targeting these early steps of oligomerization is one of the main therapeutic approaches in the quest to develop disease-modifying agents. Molecular chaperones, molecules that can mediate the proper folding and refolding of client proteins, are vital to cell function and survival and thus have been explored as potential therapeutic agents. Important to Parkinson disease, chaperones are capable of preventing α-synuclein misfolding, oligomerization, and aggregate formation as shown in vitro and in Parkinson disease animal models. Furthermore, chaperones and associated co-chaperones are closely linked to pathways of protein degradation, like the ubiquitin-proteasome system and autophagy, and are thus able to remove irreversibly misfolded proteins. In this review, we summarize the role of molecular chaperones in Parkinson disease models and discuss the importance of preserving protein homeostasis to prevent neurodegeneration. We also review the growing number of exciting studies that have targeted molecular chaperone function as a novel therapeutic approach.


Diffusion abnormality of deep gray matter in external capsular hemorrhage.
To our knowledge, diffusion abnormality of the unaffected deep gray matter during striatocapsular hemorrhage has not been previously described in the literature. We report the presence of the diffusion abnormality separated from hematoma in patients with external capsular (lateral striatocapsular) hemorrhage and suggest the plausible mechanisms of diffusion signal intensity change. We retrospectively reviewed MR images in 28 consecutive patients with spontaneous striatocapsular hemorrhage and evaluated signal intensity changes at sites separated from the hemorrhage and the lesions on diffusion-weighted (DW) images. Apparent diffusion coefficients (ADCs) of the lesions were measured, and volume changes in the deep gray matter were assessed at follow-up. On DW images, hyperintensity of deep gray matter was found in nine patients (25%). In all patients with DW imaging abnormality, the hemorrhage was located in the external capsule, and the interval from hemorrhagic ictus to MR imaging study was 8-54 days. Hyperintensity of the deep gray matter was seen in the caudate (n = 8), putamen (n = 7), thalamus (n = 5), and substantia nigra (n = 2). Mean relative ADC ratios of the diffusion abnormality were 0.76 +/- 0.10 in the caudate, 0.79 +/- 0.07 in the putamen, and 0.85 +/- 0.11 in the thalamus. DW imaging abnormality disappeared with mild atrophy in two patients who underwent follow-up imaging. External capsular hemorrhage may be uncommonly accompanied by diffusion abnormality in the striatum or thalamus at follow-up, and the lesion should not be misdiagnosed as new-onset infarction. Secondary neuronal degeneration may play an important role in the development of diffusion abnormality.


[Effect of thyroxine on the diurnal rhythm of the mitotic activity and parameters of the life cycle of cells of the liver and esophagus].
The duration of the cellular cycle and the diurnal rhythm of the amount of mitosis were studied in young rats in normality and under the influence of thyroxin. The parenchymal and connective-tissue cells of the liver and cells of the liver and the cells of the oesophagus epithelium basal layer were studied. It was found that under the influence of thyroxin there occured a shortening of the periods of the cellular cycle and a 3--6 h shift to the left of the diurnal rhythm curve of the amount of mitoses. In thyroxinized animals the 21--95% increase of the amount of mitoses in the period of maximum values of the mitotic index during a day was observed as compared with control animals. A conclusion is made about the diurnal rhythm of sensitivity of G0-phase cells to the synchronizing factor, suggesting the decisive significance of the state of the cell population in the interaction of the tissue and hormone cells. The data obatained in the work show that the thyroid hormones regulate the cellular reproduction in the organism by stimulating the cells in the division cycle, synchronization of greater amount of cells by the moment of beginning of the mitotic cycle at a definite time of day and by shortening the period of the cell mitotic cycle.


Dipeptidyl peptidase IV inhibition delays developmental programming of obesity and metabolic disease in male offspring of obese mothers.
Maternal obesity programs the offspring to metabolic diseases later in life; however, the mechanisms of programming are yet unclear, and no strategies exist for addressing its detrimental transgenerational effects. Obesity has been linked to dipeptidyl peptidase IV (DPPIV), an adipokine, and treatment of obese individuals with DPPIV inhibitors has been reported to prevent weight gain and improve metabolism. We hypothesized that DPPIV plays a role in maternal obesity-mediated programming. We measured plasma DPPIV activity in human maternal and cord blood samples from normal-weight and obese mothers at term. We found that maternal obesity increases maternal and cord blood plasma DPPIV activity but only in male offspring. Using two non-human primate models of maternal obesity, we confirmed the activation of DPPIV in the offspring of obese mothers. We then created a mouse model of maternal high-fat diet (HFD)-induced obesity, and found an early-life increase in plasma DPPIV activity in male offspring. Activation of DPPIV preceded the progression of obesity, glucose intolerance and insulin resistance in male offspring of HFD-fed mothers. We then administered sitagliptin, DPPIV inhibitor, to regular diet (RD)- and HFD-fed mothers, starting a week prior to breeding and continuing throughout pregnancy and lactation. We found that sitagliptin treatment of HFD-fed mothers delayed the progression of obesity and metabolic diseases in male offspring and had no effects on females. Our findings reveal that maternal obesity dysregulates plasma DPPIV activity in males and provide evidence that maternal inhibition of DPPIV has potential for addressing the transgenerational effects of maternal obesity.


"Pure" traumatic cerebral lacerations. A review of 129 cases with long-term follow-up.
The authors present a series of 129 patients with "pure" traumatic cerebral lacerations, i.e. not associated with other intracranial lesions (especially haematomas) which by their presence alone can play a primary role in affecting clinical course and prognosis. Twenty per cent of the patients presented a lucid interval, thus confirming the nature of the laceration as a true expanding lesion, increasing in size in the post-traumatic period. Age, GCS score and neuroradiological findings were investigated in view of the eventual outcome. Follow-up examination confirmed that age and level of coma on admission, together with the degree of ventricular shift and presence of bilateral skull fractures, were all factors of statistical significance in affecting prognosis. Lacerations in the temporal area were associated with the poorest prognosis, and highest incidence of neurological focal deficits and residual parenchymal damage on CT control.


Direct and indirect association of the small GTPase ran with nuclear pore proteins and soluble transport factors: studies in Xenopus laevis egg extracts.
Ran is a small GTPase that is required for protein import, mRNA export, and the maintenance of nuclear structures. To gain a better understanding of Ran's role in the nucleus, we have sought to use Xenopus egg extracts for the purification and characterization of proteins from egg extracts bound with a high affinity to a glutathione-S-transferase-Ran fusion protein (GST-Ran). We found that GST-Ran associates specifically with at least 10 extract proteins. We determined the identifies of six Ran-interacting proteins (Rips), and found that they include RanBP2/Nup358, Nup153, Importin beta, hsc70, RCC1, and RanBP1. On the basis of peptide sequence, a seventh Rip (p88) seems to be similar but not identical to Fug1/RanGAP1, the mammalian Ran-GTPase-activating protein. Gel filtration analysis of endogenous extract proteins suggests that Importin beta acts as a primary GTP-Ran effector. Both Ran and Importin beta are coimmunoprecipitated by anti-p340RanBP2 antibodies in the presence of nonhydrolyzable GTP analogues, suggesting that Ran-Importin beta complexes interact with p340RanBP2. Two other Rips, p18 and p88, are coprecipitated with p340RanBP2 in a nucleotide-independent manner. Analysis of the Ran-GTPase pathway in Xenopus extracts allows the examination of interactions between Ran-associated proteins under conditions that resemble in vivo conditions more closely than in assays with purified components, and it thereby allows additional insights into the molecular mechanism of nuclear transport.


Toxicity evaluation of the landfill leachate after treatment with photo-Fenton, biological and photo-Fenton followed by biological processes.
The objective of this study was the toxicity evaluation of landfill leachate samples both raw and after treatment by photo-Fenton (PF), biological (Bio) and photo-Fenton followed by biological (PF-Bio) processes. The organisms Artemia salina and Lactuca sativa were exposed to the leachate in different dilution. In the phytotoxicity evaluation, the inhibition of root (RGI) and radicle (RAGI) growth, and the number of germinated seeds (GR) were evaluated. In the ecotoxicity analysis, the 50% lethal dose (LD50) for each type of treatment was evaluated. The raw leachate, and after treatments PF, Bio and PF-Bio presented COD of 12,797, 1,347, 1,685, 220 mg/L and BOD5 of 4,251, 1,060, 692 and 89 mg/L, respectively. The GR was 0, 95, 25 and 40%, RGI was 100, 83, 100, 93%, whereas RAGI was 100, 72, 100 and 79% in the raw leachate and after treatments PF, Bio and PF-Bio, respectively. LD50 values of 0.25, 0.40, 0.38 and 0.54 were obtained for the raw effluent and after the PF, Bio and PF-Bio treatments, respectively. Smaller values of COD, BOD5, ecotoxicity and phytotoxicity were observed in the effluent after the PF-Bio treatment, indicating that this is the most environmentally suitable system for the treatment of landfill leachate.


Predictors of supportive message expression and reception in an interactive cancer communication system.
Social support in computer-mediated settings is an important variable in health communication research, yet little is known about the factors that influence the amount of social support one gives and receives in online support groups. To shed some light on this issue, the authors examined demographics, disease-related factors, psychosocial factors, and strategies for coping with breast cancer as potential determinants of which patients provide support to others and which ones consume it. Data collected from 177 participants in the Comprehensive Health Enhancement Support System "Living With Breast Cancer" program revealed that individuals who are younger, have higher levels of positive reframing, and lower levels of self-blame are more likely to provide emotional support in online settings. In contrast, individuals who are more educated, have less perceived availability of social support, and have lower levels of religious coping are more likely to receive emotional support from others. The authors discuss the theoretical and practical implications for providing effective psychosocial support for women with breast cancer.


Anti-ageing effects of a new synthetic sphingolipid (K6EAA-L12) on aged murine skin.
Recently, we reported on the anti-ageing effects of K6PC-5. This compound induced keratinocyte differentiation and fibroblast proliferation by increasing sphingosine-1 phosphate synthesis. We performed this study to confirm the anti-ageing effects of new synthetic products (the K6EAA series) derived from K6PC-5 through an amino group induction. Cellular responses such as differentiation, proliferation and calcium mobilization were investigated using cultured human keratinocytes and fibroblasts. Also, we measured the expressions of collagen mRNA and protein using real time RT-PCR and ELISA, respectively. The K6EAA-L12 product, selected by in vitro screening, was evaluated for anti-ageing effects on intrinsically and extrinsically (photo) aged models of hairless mice. In the intrinsically aged murine skin, K6EAA-L12 showed anti-ageing effects by activating collagen synthesis, eventually causing dermal thickening. Also, in the photo-aged skin, the dermal collagen density and dermal thickness were increased. In photo-aged murine skin, K6EAA-L12 increased stratum corneum integrity by increasing corneodesmosome density and improved the barrier recovery rate. However, there were no changes in the expressions of epidermal differentiation maker proteins. In conclusion, topical K6EAA-L12, a new synthetic K6PC-5 derivative, improves intrinsically and extrinsically (photo) aged skin by increasing the collagen density and improving the skin barrier function.


Consensus models to predict oral rat acute toxicity and validation on a dataset coming from the industrial context.
We report predictive models of acute oral systemic toxicity representing a follow-up of our previous work in the framework of the NICEATM project. It includes the update of original models through the addition of new data and an external validation of the models using a dataset relevant for the chemical industry context. A regression model for LD50 and multi-class classification model for toxicity classes according to the Global Harmonized System categories were prepared. ISIDA descriptors were used to encode molecular structures. Machine learning algorithms included support vector machine (SVM), random forest (RF) and naïve Bayesian. Selected individual models were combined in consensus. The different datasets were compared using the generative topographic mapping approach. It appeared that the NICEATM datasets were lacking some relevant chemotypes for chemical industry. The new models trained on enlarged data sets have applicability domains (AD) sufficiently large to accommodate industrial compounds. The fraction of compounds inside the models' AD increased from 58% (NICEATM model) to 94% (new model). The increase of training sets improved models' prediction performance: RMSE values decreased from 0.56 to 0.47 and balanced accuracies increased from 0.69 to 0.71 for NICEATM and new models, respectively.


Loneliness and its association with physical health conditions and psychiatric hospitalizations in people with serious mental illness.
Explore the association between loneliness and psychiatric hospitalizations, functioning, and physical health conditions in people with serious mental illness. Data were included from 349 individuals with serious mental illnesses, aged 18+ years, who participated in two consecutive cross-sectional survey studies conducted between 2013 and 2018. A higher number of psychiatric hospitalizations in the past six months was associated with greater loneliness. The Short-Form-12 mental component score was a highly significant correlate, accounting for a significant amount of variance in loneliness scores. The physical component score and the bodily pain subdomain of the Short-Form-12 were also significantly associated with loneliness, with individuals with lower physical functioning and greater bodily pain reporting more loneliness. Having a higher total number of health conditions, higher total number of conditions that cause pain, and the presence of at least one respiratory condition were found to be associated with greater loneliness. Loneliness may be an important dimension of health as related to health conditions associated with pain, functioning, and psychiatric hospitalizations among people with serious mental illness. These finding reinforce the powerful role of socio-cultural and environmental factors in shaping whole health in people with serious mental illness.


Protein QTL analysis of IGF-I and its binding proteins provides insights into growth biology.
The growth hormone and insulin-like growth factor (IGF) system is integral to human growth. Genome-wide association studies (GWAS) have identified variants associated with height and located near the genes in this pathway. However, mechanisms underlying these genetic associations are not understood. To investigate the regulation of the genes in this pathway and mechanisms by which regulation could affect growth, we performed GWAS of measured serum protein levels of IGF-I, IGF binding protein-3 (IGFBP-3), pregnancy-associated plasma protein A (PAPP-A2), IGF-II and IGFBP-5 in 838 children (3-18 years) from the Cincinnati Genomic Control Cohort. We identified variants associated with protein levels near IGFBP3 and IGFBP5 genes, which contain multiple signals of association with height and other skeletal growth phenotypes. Surprisingly, variants that associate with protein levels at these two loci do not colocalize with height associations, confirmed through conditional analysis. Rather, the IGFBP3 signal (associated with total IGFBP-3 and IGF-II levels) colocalizes with an association with sitting height ratio (SHR); the IGFBP5 signal (associated with IGFBP-5 levels) colocalizes with birth weight. Indeed, height-associated single nucleotide polymorphisms near genes encoding other proteins in this pathway are not associated with serum levels, possibly excluding PAPP-A2. Mendelian randomization supports a stronger causal relationship of measured serum levels with SHR (for IGFBP-3) and birth weight (for IGFBP-5) than with height. In conclusion, we begin to characterize the genetic regulation of serum levels of IGF-related proteins in childhood. Furthermore, our data strongly suggest the existence of growth-regulating mechanisms acting through IGF-related genes in ways that are not reflected in measured serum levels of the corresponding proteins.


Terminal alkenes as versatile chemical reporter groups for metabolic oligosaccharide engineering.
The Diels-Alder reaction with inverse electron demand (DAinv reaction) of 1,2,4,5-tetrazines with electron rich or strained alkenes was proven to be a bioorthogonal ligation reaction that proceeds fast and with high yields. An important application of the DAinv reaction is metabolic oligosaccharide engineering (MOE) which allows the visualization of glycoconjugates in living cells. In this approach, a sugar derivative bearing a chemical reporter group is metabolically incorporated into cellular glycoconjugates and subsequently derivatized with a probe by means of a bioorthogonal ligation reaction. Here, we investigated a series of new mannosamine and glucosamine derivatives with carbamate-linked side chains of varying length terminated by alkene groups and their suitability for labeling cell-surface glycans. Kinetic investigations showed that the reactivity of the alkenes in DAinv reactions increases with growing chain length. When applied to MOE, one of the compounds, peracetylated N-butenyloxycarbonylmannosamine, was especially well suited for labeling cell-surface glycans. Obviously, the length of its side chain represents the optimal balance between incorporation efficiency and speed of the labeling reaction. Sialidase treatment of the cells before the bioorthogonal labeling reaction showed that this sugar derivative is attached to the glycans in form of the corresponding sialic acid derivative and not epimerized to another hexosamine derivative to a considerable extent.


Netschaevo: a new class of chondritic meteorite.
The ratios of refractory elements to silicon and of zinc to silicon indicate that the silicate portion of the Netschaëvo meteorite is an ordinary chondrite. The scarcity of chondrules, the large dimensions (about 100 micrometers) of plagioclase grains, and the low indium content (0.09 nanogram per gram) indicate that Netschaëvo belongs to petrologic type 6. On a diagram of reduced iron versus oxidized iron, Netschaëvo lies along an extrapolation of the LL-L-H ordinary chondrite fractionation trend. The abundances of siderophile elements (nickel, germanium, iridium, and gold) are about 1.6 to 2.0 times greater than in H-group chondrites, and siderophile/nickel ratios are, with one exception, those expected from LL-L-H trends. This evidence indicates that Netschaëvo is an extremely iron-rich member of the ordinary chondrite sequence, and that plausible models to account for the ordinary chondrite sequence must produce materials having iron/silicon ratios 25 percent greater than those in CI carbonaceous chondrites. The existence of Netschaëvo emphasizes that the chondritic meteorites in terrestrial collections are a biased and incomplete selection of primitive solar system materials.


Myogenic precursor cells in craniofacial muscles.
Craniofacial skeletal muscles (CskM), including the masticatory (MM), extraocular (EOM) and laryngeal muscles (LM), have a number of properties that set them apart from the majority of skeletal muscles (SkM). They have embryological origins that are distinct from musculature elsewhere in the body, they express a number of immature myosin heavy chain isoforms and maintain increased and distinct expression of a number of myogenic growth factors and their receptors from other adult SkMs. Furthermore, it has recently been demonstrated that unlike limb SkM, normal adult EOM and LM retain a population of activated satellite cells, the regenerative cell in adult SkM. In order to maintain this proliferative pool throughout life, CSkM may contain more satellite cells and/or more multipotent precursor cells that may be more resistant to apoptosis than those found in limb muscle. A further exciting question is whether this potentially more active muscle precursor cell population could be utilized not only for SkM repair, but be harnessed for repair or reconstruction of other tissues, such as nervous tissue or bone. This is a highly attractive speculation as the innate regenerative capacity of craniofacial muscles would ensure the donor tissue would not have compromised future function.


A raised serum lactate level is an independent predictor of in-hospital mortality in patients with isolated cerebral gunshot wounds.
Cerebral gunshot wounds (CGSWs) represent a highly lethal form of traumatic brain injury, and triaging these patients is difficult. The prognostic significance of the serum lactate level in the setting of CGSWs is largely unknown. To examine the relationship between elevated serum lactate levels and mortality in patients with isolated CGSWs. A retrospective review of the regional trauma registry was undertaken at the Pietermaritzburg Metropolitan Trauma Service, South Africa, over a 5-year period from 1 January 2010 to 31 December 2014. All patients with an isolated CGSW were included. A total of 102 patients with isolated CGSWs were identified. Of these, 92.2% (94/102) were male. The mean age (standard deviation) was 29 (8) years, and the in-hospital mortality rate was 21.6% (22/102). The mean serum lactate level was significantly higher among non-survivors than among survivors (6.1 mmol/L v. 1.3 mmol/L; p<0.001). Lactate levels among non-survivors were <2 mmol/L in 4.5%, 2 - 3.99 mmol/L in 9.1%, 4 - 5.99 mmol/L in 36.4% and ≥6 mmol/L in 50.0%. The odds ratio for mortality with a lactate level of 4 - 5.99 mmol/L was 67 (95% confidence interval (CI) 1.7 - 2 674.2), while for a lactate level of ≥6 mmol/L it was 1 787 (95% CI 9.0 - 354 116.1). The serum lactate level accurately predicted mortality even after adjustment for other variables. Based on a receiver operating curve analysis, an optimal cut-off of 3.3 mmol/L for serum lactate as a predictor for mortality was identified (area under the curve = 0.957). CGSWs are associated with significant mortality, and a raised serum lactate level appears to be an independent predictor of in-hospital mortality. It is a potentially useful adjunct in the resuscitation room for identifying patients with a very poor prognosis.


All-optical control of gigahertz acoustic resonances by forward stimulated interpolarization scattering in a photonic crystal fiber.
We report the observation of a novel nonlinear optoacoustic phenomenon, that we name forward stimulated interpolarization scattering. When two frequency-offset laser signals are colaunched into orthogonally polarized guided modes of a birefringent small-core (1.8 μm diameter) photonic crystal fiber, a pattern of axially moving polarization fringes is produced, with a velocity and spacing that depends on the frequency offset. At values of frequency offset in the few-GHz range, the pattern of moving fringes can perfectly match the phase velocity and axial wavelength (3.9 mm) of the torsional-radial acoustic mode tightly guided in the core. An intense optoacoustic interaction ensues, leading to efficient nonlinear exchange of power from the higher frequency (pump) mode to the orthogonally polarized lower frequency (Stokes) mode. A full-vectorial theory is developed to explain the observations.


Exome chip analyses identify genes affecting mortality after HLA-matched unrelated-donor blood and marrow transplantation.
Although survival outcomes have significantly improved, up to 40% of patients die within 1 year of HLA-matched unrelated-donor blood and marrow transplantation (BMT). To identify non-HLA genetic contributors to mortality after BMT, we performed the first exome-wide association study in the DISCOVeRY-BMT cohorts using the Illumina HumanExome BeadChip. This study includes 2473 patients with acute myeloid leukemia, acute lymphoblastic leukemia, or myelodysplastic syndrome and 2221 10/10 HLA-matched donors treated from 2000 to 2011. Single-variant and gene-level analyses were performed on overall survival (OS), transplantation-related mortality (TRM), and disease-related mortality (DRM). Genotype mismatches between recipients and donors in a rare nonsynonymous variant of testis-expressed gene TEX38 significantly increased risk of TRM, which was more dramatic when either the recipient or donor was female. Using the SKAT-O test to evaluate gene-level effects, variant genotypes of OR51D1 in recipients were significantly associated with OS and TRM. In donors, 4 (ALPP, EMID1, SLC44A5, LRP1), 1 (HHAT), and 2 genes (LYZL4, NT5E) were significantly associated with OS, TRM, and DRM, respectively. Inspection of NT5E crystal structures showed 4 of the associated variants affected the enzyme structure and likely decreased the catalytic efficiency of the enzyme. Further confirmation of these findings and additional functional studies may provide individualized risk prediction and prognosis, as well as alternative donor selection strategies.


Pyruvate carboxylation prevents the decline in contractile function of rat hearts oxidizing acetoacetate.
Acetoacetate, when present as the only fuel for respiration in rat hearts, causes an impairment in contractile function that is reversible with the addition of substrates that can contribute to anaplerosis. To determine the importance of pyruvate carboxylation via NADP(+)-dependent malic enzyme on metabolism and function in hearts oxidizing acetoacetate, isolated working rat hearts were perfused with [1-14C]pyruvate and acetoacetate. While the cardiac power output after 60 min of perfusion in hearts utilizing acetoacetate alone had fallen to 44% of the initial value, the addition of pyruvate resulted in a stable performance with no fall in the work output. When hydroxymalonate, an inhibitor of NADP(+)-dependent malic enzyme and malate dehydrogenase, was added to the two substrates, function at 60 min was similar to the value for hearts oxidizing acetoacetate alone. Measurements of the specific activities of malate, aspartate, and citrate confirm inhibition of both pyruvate carboxylation and malate oxidation. The findings are consistent with a mechanism in which the enrichment of malate by pyruvate improves function by increasing the production of reducing equivalents by the malate dehydrogenase and the isocitrate dehydrogenase reactions increase flux through the span of the tricarboxylic acid cycle from malate to 2-oxoglutarate. The present study demonstrates the physiological importance of anaplerotic pathways in maintaining contractile function in the heart.


Discretionary decision making by primary care physicians and the cost of U.S. Health care.
Efforts to improve the quality and costs of U.S. health care have focused largely on fostering physician adherence to evidence-based guidelines, ignoring the role of clinical judgment in more discretionary settings. We surveyed primary care physicians to assess variability in discretionary decision making and evaluate its relationship to the cost of health care. Physicians in high-spending regions see patients back more frequently and are more likely to recommend screening tests of unproven benefit and discretionary interventions compared with physicians in low-spending regions; however, both appear equally likely to recommend guideline-supported interventions. Greater attention should be paid to the local factors that influence physicians' clinical judgment in discretionary settings.


Therapeutic results of intra-arterial thrombolysis after full-dose intravenous tissue plasminogen activator administration.
IV administration of tPA is accepted as a standard treatment for acute cerebral ischemia, but the clinical outcomes cannot be guaranteed in patients who are not recanalized after IV-tPA and in those who are not eligible for IV-tPA. In this study, outcomes from groups of patients treated with additional IA thrombolytic therapy with the use or omission of IV-tPA administration were compared. IA thrombolytic therapy (thrombolytic agents combined with mechanical intervention) was attempted in those patients who were not eligible for IV-tPA and who showed continuous major vessel occlusion after IV-tPA. Sixty-three patients were divided into 2 groups: a tPA group (n = 29, IA thrombolysis after IV-tPA) and a non-tPA group (n = 34, IA thrombolysis without IV-tPA). These groups were subdivided according to match or mismatch DWI/PWI after MR imaging. Treatment results were compared by recanalization rate, clinical outcome, mortality, and ICH rate. The recanalization rate was 79.3% in the tPA group and 55.9% in the non-tPA group (χ(2) test, P < .05). Subgroup analysis between DWI/PWI mismatch in the tPA group and DWI/PWI mismatch in the non-tPA group also showed no statistical difference in recanalization rate, favorable clinical outcome, and mortality (χ(2) test, P > .05), but the significant ICH rate was high in the tPA group (χ(2) test, P < .05). Additional IA thrombolytic treatment after full-dose IV-tPA administration might be an acceptable treatment option for patients with DWI/PWI mismatch.


Diagnostic Accuracy of Coronary CT Angiography for the Evaluation of Bioresorbable Vascular Scaffolds.
The purpose of this study was to assess the diagnostic accuracy of coronary computed tomography angiography (CTA) for bioresorbable vascular scaffold (BVS) evaluation. Coronary CTA has emerged as a noninvasive method to evaluate patients with suspected or established coronary artery disease. The diagnostic accuracy of coronary CTA to evaluate angiographic outcomes after BVS implantation has not been well established. In the ABSORB II (A Bioresorbable Everolimus-Eluting Scaffold Versus a Metallic Everolimus-Eluting Stent II) study, patients were randomized either to receive treatment with the BVS or everolimus-eluting metallic stent. At the 3-year follow-up, 238 patients (258 lesions) treated with BVS underwent coronary angiography with intravascular ultrasound (IVUS) evaluation and coronary CTA. The diagnostic accuracy of coronary CTA was assessed by the area under the receiver-operating characteristic curve with coronary angiography and IVUS as references. The mean difference in coronary CTA-derived minimal luminal diameter was -0.14 mm (limits of agreement -0.88 to 0.60) with quantitative coronary angiography as reference, whereas the mean difference in minimal lumen area was 0.73 mm2 (limits of agreement -1.85 to 3.30) with IVUS as reference. The per-scaffold diagnostic accuracy of coronary CTA for detecting stenosis based on coronary angiography diameter stenosis of ≥50% revealed an area under the receiver-operating characteristic curve of 0.88 (95% confidence interval [CI]: 0.82 to 0.92) with a sensitivity of 80% (95% CI: 28% to 99%) and a specificity of 100% (95% CI: 98% to 100%), whereas diagnostic accuracy based on IVUS minimal lumen area ≤2.5 mm2 showed an area under the receiver-operating characteristic curve of 0.83 (95% CI: 0.77 to 0.88) with a sensitivity of 71% (95% CI: 44% to 90%) and a specificity of 82% (95% CI: 75% to 87%). The diagnostic accuracy of coronary CTA was similar to coronary angiography in its ability to identify patients with a significant lesion based on the IVUS criteria (p = 0.75). Coronary CTA has good diagnostic accuracy to detect in-scaffold luminal obstruction and to assess luminal dimensions after BVS implantation. Coronary angiography and coronary CTA yielded similar diagnostic accuracy to identify the presence and severity of obstructive disease. Coronary CTA might become the method of choice for the evaluation of patients treated with BVS.


Evolutionarily stable infection by a male-killing endosymbiont in Drosophila innubila: molecular evidence from the host and parasite genomes.
Maternally inherited microbes that spread via male-killing are common pathogens of insects, yet very little is known about the evolutionary duration of these associations. The few examples to date indicate very recent, and thus potentially transient, infections. A male-killing strain of Wolbachia has recently been discovered in natural populations of Drosophila innubila. The population-level effects of this infection are significant: approximately 35% of females are infected, infected females produce very strongly female-biased sex ratios, and the resulting population-level sex ratio is significantly female biased. Using data on infection prevalence and Wolbachia transmission rates, infected cytoplasmic lineages are estimated to experience a approximately 5% selective advantage relative to uninfected lineages. The evolutionary history of this infection was explored by surveying patterns of polymorphism in both the host and parasite genomes, comparing the Wolbachia wsp gene and the host mtDNA COI gene to five host nuclear genes. Molecular data suggest that this male-killing infection is evolutionarily old, a conclusion supported with a simple model of parasite and mtDNA transmission dynamics. Despite a large effective population size of the host species and strong selection to evolve resistance, the D. innubila-Wolbachia association is likely at a stable equilibrium that is maintained by imperfect maternal transmission of the bacteria rather than partial resistance in the host species.


Effects of stimulus predictability and interstimulus gap on saccades in Alzheimer's disease.
Studies of saccadic eye movement impairment in Alzheimer's disease (AD) have largely focused on simple reflexive eye movements and the antisaccade task. The effects of manipulating stimulus timing have been little studied. Fourteen patients with mild to severe AD and 11 age-matched controls were studied on the antisaccade task, while latencies on simultaneous, gap and predictable tasks were recorded from 11 patients and 11 controls. Dementia severity was assessed with the Mini-Mental State Examination. As a group, patients' latencies were significantly higher and more variable on the simultaneous and gap tasks. Predictable task performance was similar in mean but significantly more variable. Grossly anticipatory responses by patients were common on the predictable, simultaneous and gap tasks. Exclusion of these from subject means revealed that AD patients, when making target-driven saccades, demonstrated a gap effect of similar magnitude to normal subjects. Patients made significantly fewer correct antisaccades and significantly more reflexive errors not followed by a corrective antisaccade than did controls. The frequent presence of grossly anticipatory saccades may reflect dysfunction of fixation mechanisms possibly involving projections from frontal lobe to superior colliculus. The less frequently seen, marked prolongation of latency may reflect changes in posterior parietal mechanisms mediating reflexive saccade generation. The presence of the gap effect demonstrates a continued ability to benefit from externally controlled stimulus disengagement. Patients' ability to make appropriately timed saccades to targets of known locations was particularly impaired, but the target sequence itself was at least grossly correctly learned. Larger studies may be able to identify clinically distinct populations of AD patients.


Gene-environment interaction in atopic diseases: a population-based twin study of early-life exposures.
The development of atopic diseases early in life suggests an important role of perinatal risk factors. To study whether early-life exposures modify the genetic influence on atopic diseases in a twin population. Questionnaire data on atopic diseases from 850 monozygotic and 2279 like-sex dizygotic twin pairs, 3-9 years of age, from the Danish Twin Registry were cross-linked with data on prematurity, Cesarean section, maternal age at birth, parental cohabitation, season of birth and maternal smoking during pregnancy, from the Danish National Birth Registry. Significant predictors of atopic diseases were identified with logistic regression and subsequently tested for genetic effect modification using variance components analysis. After multivariable adjustment, prematurity (gestational age below 32 weeks) [odds ratio (OR) = 1.93, confidence interval (CI) = 1.45-2.56], Cesarean section (OR = 1.25, CI = 1.05-1.49) and maternal smoking during pregnancy (OR = 1.70, CI = 1.42-2.04) significantly influenced the risk of asthma, whereas none of the factors were significantly associated with atopic dermatitis and hay fever. Variance components analysis stratified by exposure status showed no significant change in the heritability of asthma according to the identified risk factors. In this population-based study of children, there was no evidence of genetic effect modification of atopic diseases by several identified early-life risk factors. The causal relationship between these risk factors and atopic diseases may therefore be mediated via mechanisms different from gene-environment interaction.


Sex differences in assessment of obesity in rheumatoid arthritis.
To determine the prevalence of obesity and how accurately standard anthropometric measures identify obesity among men and women with rheumatoid arthritis (RA). Dual x-ray absorptiometry (DXA) was performed for 141 persons with RA (56 men and 85 women). Two anthropometric proxies of obesity (body mass index [BMI] and waist circumference [WC]) were compared to a DXA-based obesity criterion. Receiver operating characteristic curves determined optimal cut points for each anthropometric measure, relative to DXA. The association of body fat and anthropometric obesity measures with disease status and cardiovascular risk was assessed in multiple regression analyses, controlling for age and glucocorticoid use. All analyses were performed separately for men and women. A total of 20%, 32%, and 44% of women and 41%, 36%, and 80% of men were classified as obese by BMI, WC, and DXA, respectively. Cut points were identified for anthropometric measures to better approximate DXA estimates of percent body fat (BMI ≥26.1 kg/m(2) for women and ≥24.7 kg/m(2) for men; WC ≥83 cm for women and ≥96 cm for men). For women and men, higher percent fat was associated with poorer RA status. Anthropometric measures were more closely linked to RA status for women, but identified cardiovascular risk for both women and men. A large percentage of this RA sample was overfat; DXA-defined obesity was twice as common in men as in women. Utility of revised BMI and WC cut points compared to traditional cut points remains to be examined in prospective studies, but results suggest that lower, sex-specific cut points may be warranted to better identify individuals at risk for poor RA and/or cardiovascular outcomes.


Bacterial factors that mediate colonization of the stomach and virulence of Helicobacter pylori.
Helicobacter pylori is a Gram-negative microaerophilic organism that colonizes the gastric mucosa of humans. Helicobacter pylori is one of the most common infections in humans and results in the development of gastritis in all infected individuals, although the majority of people are asymptomatic. A subset of infected people develop serious disease including duodenal ulceration and gastric cancer. Helicobacter pylori exhibits many striking characteristics. It lives in the hostile environment of the stomach and displays a very strict host and tissue tropism. Despite a vigorous immune response, infection persists for the lifetime of the host unless eradicated with antimicrobials. Why H. pylori is so pathogenic in some individuals and not in others is unknown but is thought to be due to a variety of host, environmental and bacterial factors. In this review, some of the bacterial factors that mediate colonization of the gastric mucosa and play a role in the pathogenesis of this organism have been considered.


Neurogenic inflammation and the peripheral nervous system in host defense and immunopathology.
The peripheral nervous and immune systems are traditionally thought of as serving separate functions. The line between them is, however, becoming increasingly blurred by new insights into neurogenic inflammation. Nociceptor neurons possess many of the same molecular recognition pathways for danger as immune cells, and, in response to danger, the peripheral nervous system directly communicates with the immune system, forming an integrated protective mechanism. The dense innervation network of sensory and autonomic fibers in peripheral tissues and high speed of neural transduction allows rapid local and systemic neurogenic modulation of immunity. Peripheral neurons also seem to contribute to immune dysfunction in autoimmune and allergic diseases. Therefore, understanding the coordinated interaction of peripheral neurons with immune cells may advance therapeutic approaches to increase host defense and suppress immunopathology.


Coherent feedback control of a single qubit in diamond.
Engineering desired operations on qubits subjected to the deleterious effects of their environment is a critical task in quantum information processing, quantum simulation and sensing. The most common approach relies on open-loop quantum control techniques, including optimal-control algorithms based on analytical or numerical solutions, Lyapunov design and Hamiltonian engineering. An alternative strategy, inspired by the success of classical control, is feedback control. Because of the complications introduced by quantum measurement, closed-loop control is less pervasive in the quantum setting and, with exceptions, its experimental implementations have been mainly limited to quantum optics experiments. Here we implement a feedback-control algorithm using a solid-state spin qubit system associated with the nitrogen vacancy centre in diamond, using coherent feedback to overcome the limitations of measurement-based feedback, and show that it can protect the qubit against intrinsic dephasing noise for milliseconds. In coherent feedback, the quantum system is connected to an auxiliary quantum controller (ancilla) that acquires information about the output state of the system (by an entangling operation) and performs an appropriate feedback action (by a conditional gate). In contrast to open-loop dynamical decoupling techniques, feedback control can protect the qubit even against Markovian noise and for an arbitrary period of time (limited only by the coherence time of the ancilla), while allowing gate operations. It is thus more closely related to quantum error-correction schemes, although these require larger and increasing qubit overheads. Increasing the number of fresh ancillas enables protection beyond their coherence time. We further evaluate the robustness of the feedback protocol, which could be applied to quantum computation and sensing, by exploring a trade-off between information gain and decoherence protection, as measurement of the ancilla-qubit correlation after the feedback algorithm voids the protection, even if the rest of the dynamics is unchanged.


Effectiveness and strategies of iron supplementation during pregnancy.
Iron deficiency continues to be one of the most prevalent single-nutrient deficiencies in the world. Interventions are often designed to prevent the decrease in hemoglobin concentration and the decline in iron stores associated with pregnancy. Although this is believed to be desirable for both the health of the mother and the well-being of the growing fetus, some scientists disagree. Enrichment and fortification of food items, and dietary changes resulting from education interventions, have met with some success in developed countries, but not often in the developing world. A therapeutic approach to iron supplementation, rather than a public health-based approach, is used throughout much of the world but suffers from real, or perceived, problems of compliance. Large doses of iron are most often prescribed and are associated with side effects and with increased oxidative damage. Alternatively, delayed-release preparations and intermittent oral iron supplementation lead to better overall compliance and alleviate side effects. Daily iron intervention provides more protection against a decline in the storage iron pool in pregnant women than does an intermittent schedule, but the latter is generally associated with fewer side effects, better compliance, and possibly a reduction in risk of oxidative damage. An improved cost-benefit ratio associated with a lower-dose oral iron supplement may prove to be quite positive in the future. Currently, no single approach may be universally acceptable, although a moderate iron dosage protocol will likely provide the most benefit to those who require supplemental iron.


EQ-5D-5L in the General German Population: Comparison and Evaluation of Three Yearly Cross-Section Surveys.
Health-related quality of life (HRQoL) is a key measure for evaluating health status in populations. Using the recent EQ-5D-5L for measurement, this study analyzed quality of life results and their stability over consecutive population surveys. Three cross-section surveys for representative samples of the general German population from 2012, 2013, and 2014 were evaluated using the EQ-5D-5L descriptive system and valuation by the Visual Analog Scale (VAS). Aggregated sample size reached 6074. The dimension with the highest prevalence of problems was pain/discomfort (31.7%). Compared with 2012 (59.3%), the percentage of participants in the best health state increased slightly in 2013 (63.4%) and 2014 (62%). Over the 3-year period, diabetes and heart disease had the strongest negative influence on mean VAS result. The number of reported chronic diseases cumulatively reduced mean VAS. Extreme problems in one or more dimensions were stated by only 0.1%-0.2% of patients. Of the potential 247 health states with a problem score ≥ 20, only six were observed in the aggregated sample. HRQoL results were fairly stable over the 3 years, but the share of the population with no problems was not. Results from the aggregated sample may serve as updated reference values for the general German population.


Cell Heterogeneity and Phenotypic Plasticity in Metastasis Formation: The Case of Colon Cancer.
The adenoma-to-carcinoma progression in colon cancer is driven by a sequential accumulation of genetic alterations at specific tumor suppressors and oncogenes. In contrast, the multistage route from the primary site to metastasis formation is underlined by phenotypic plasticity, i.e., the capacity of disseminated tumor cells to undergo transiently and reversible transformations in order to adapt to the ever-changing environmental contexts. Notwithstanding the considerable body of evidence in support of the role played by epithelial-to-mesenchymal transition (EMT)/mesenchymal-to-epithelial transition (MET) in metastasis, its rate-limiting function, the detailed underlying cellular and molecular mechanisms, and the extension of the necessary morphologic and epigenetic changes are still a matter of debate. Rather than leading to a complete epithelial or mesenchymal state, the EMT/MET-program generates migrating cancer cells displaying intermediate phenotypes featuring both epithelial and mesenchymal characteristics. In this review, we will address the role of colon cancer heterogeneity and phenotypic plasticity in metastasis formation and the contribution of EMT to these processes. The alleged role of hybrid epithelial/mesenchymal (E/M) in collective and/or single-cell migration during local dissemination at the primary site and more systemic spreading will also be highlighted.


Role of PCNA and RFC in promoting Mus81-complex activity.
Proper DNA replication is essential for faithful transmission of the genome. However, replication stress has serious impact on the integrity of the cell, leading to stalling or collapse of replication forks, and has been determined as a driving force of carcinogenesis. Mus81-Mms4 complex is a structure-specific endonuclease previously shown to be involved in processing of aberrant replication intermediates and promotes POLD3-dependent DNA synthesis via break-induced replication. However, how replication components might be involved in this process is not known. Herein, we show the interaction and robust stimulation of Mus81-Mms4 nuclease activity by heteropentameric replication factor C (RFC) complex, the processivity factor of replicative DNA polymerases that is responsible for loading of proliferating cell nuclear antigen (PCNA) during DNA replication and repair. This stimulation is enhanced by RFC-dependent ATP hydrolysis and by PCNA loading on the DNA. Moreover, this stimulation is not specific to Rfc1, the largest of subunit of this complex, thus indicating that alternative clamp loaders may also play a role in the stimulation. We also observed a targeting of Mus81 by RFC to the nick-containing DNA substrate and we provide further evidence that indicates cooperation between Mus81 and the RFC complex in the repair of DNA lesions generated by various DNA-damaging agents. Identification of new interacting partners and modulators of Mus81-Mms4 nuclease, RFC, and PCNA imply the cooperation of these factors in resolution of stalled replication forks and branched DNA structures emanating from the restarted replication forks under conditions of replication stress.


MobiDB-lite: fast and highly specific consensus prediction of intrinsic disorder in proteins.
Intrinsic disorder (ID) is established as an important feature of protein sequences. Its use in proteome annotation is however hampered by the availability of many methods with similar performance at the single residue level, which have mostly not been optimized to predict long ID regions of size comparable to domains. Here, we have focused on providing a single consensus-based prediction, MobiDB-lite, optimized for highly specific (i.e. few false positive) predictions of long disorder. The method uses eight different predictors to derive a consensus which is then filtered for spurious short predictions. Consensus prediction is shown to outperform the single methods when annotating long ID regions. MobiDB-lite can be useful in large-scale annotation scenarios and has indeed already been integrated in the MobiDB, DisProt and InterPro databases. MobiDB-lite is available as part of the MobiDB database from URL: http://mobidb.bio.unipd.it/. An executable can be downloaded from URL: http://protein.bio.unipd.it/mobidblite/. email@example.com. Supplementary data are available at Bioinformatics online.


Age-related changes in the distribution of Kv1.1 and Kv3.1 in rat cochlear nuclei.
To identify age-related changes in voltage-gated K(+) (Kv) channels that contribute to temporal processing in neurons of the central auditory system, we investigated the distribution of Kv1.1 and Kv3.1 in the auditory brainstem of adult and aged rats. Immunohistochemistry was performed in accordance with the free-floating method described earlier. Among the auditory nuclei, only the posterior ventral cochlear nucleus (PVCN) showed age-related changes. Kv1.1 immunoreactivity was increased in the octopus cell bodies, while the staining intensity was significantly decreased in the neuropil. Image analysis demonstrated the specific increase in Kv1.1 immunoreactivity in aged cochlear nucleus neurons although the mean density of the entire selection was significantly decreased. In contrast, the number of Kv1.1-immunoreactive neurons was not significantly different between control and aged groups. The immunoreactivity for Kv3.1 was decreased in the octopus cells and neuropil of aged PVCN, which was confirmed by image analysis. The number of Kv3.1-positive cells was also significantly decreased in aged PVCN. This study may provide useful data to compare age-related changes in Kv1.1 and Kv3.1 with known physiological properties of auditory neurons.


Spatial relationships between lead sources and children's blood lead levels in the urban center of Indianapolis (USA).
Urban children remain disproportionately at risk of having higher blood lead levels than their suburban counterparts. The Westside Cooperative Organization (WESCO), located in Marion County, Indianapolis, Indiana, has a history of children with high blood lead levels as well as high soil lead (Pb) values. This study aims at determining the spatial relationship between soil Pb sources and children's blood lead levels. Soils have been identified as a source of chronic Pb exposure to children, but the spatial scale of the source-recipient relationship is not well characterized. Neighborhood-wide analysis of soil Pb distribution along with a furnace filter technique for sampling interior Pb accumulation for selected homes (n = 7) in the WESCO community was performed. Blood lead levels for children aged 0-5 years during the period 1999-2008 were collected. The study population's mean blood lead levels were higher than national averages across all ages, race, and gender. Non-Hispanic blacks and those individuals in the Wishard advantage program had the highest proportion of elevated blood lead levels. The results show that while there is not a direct relationship between soil Pb and children's blood lead levels at a spatial scale of ~100 m, resuspension of locally sourced soil is occurring based on the interior Pb accumulation. County-wide, the largest predictor of elevated blood lead levels is the location within the urban core. Variation in soil Pb and blood lead levels on the community level is high and not predicted by housing stock age or income. Race is a strong predictor for blood lead levels in the WESCO community.


Congenital bacterial sepsis in very preterm infants.
The results of body fluid and surface cultures from 148 preterm infants less than 33 weeks gestational age obtained routinely on admission to a neonatal intensive care unit were reviewed. The aim was to determine the occurrence of congenital bacterial sepsis in this population and to examine whether surface cultures yielded information helpful in management. Gastric aspirate and umbilical, nasal and ear swabs were cultured and the results were compared to those of blood cultures. Nine infants (5.4%) had congenital bacterial sepsis diagnosed by positive blood cultures. Only the results of microscopy of gastric aspirate were available within hours of birth and before the results of blood culture. Microscopy of gastric aspirate, demonstrating pus cells, alone had a sensitivity of 0.86 in predicting congenital sepsis but a specificity of 0.49; the specificity, however, rose to 0.80 if both organisms and pus cells were observed on microscopy. Thus, only this combination was a useful pre-indicator of congenital sepsis. In infants who did not develop septicaemia, treatment was modified only if Streptococcus agalactiae was cultured from surface sites; in all such cases, the organism was grown from the ear swab. Our results demonstrate that congenital bacterial sepsis is common amongst very preterm infants admitted for neonatal intensive care but routine screening of surface cultures should be restricted to an ear swab only.


Roles of the WHHL rabbit in translational research on hypercholesterolemia and cardiovascular diseases.
Conquering cardiovascular diseases is one of the most important problems in human health. To overcome cardiovascular diseases, animal models have played important roles. Although the prevalence of genetically modified animals, particularly mice and rats, has contributed greatly to biomedical research, not all human diseases can be investigated in this way. In the study of cardiovascular diseases, mice and rats are inappropriate because of marked differences in lipoprotein metabolism, pathophysiological findings of atherosclerosis, and cardiac function. On the other hand, since lipoprotein metabolism and atherosclerotic lesions in rabbits closely resemble those in humans, several useful animal models for these diseases have been developed in rabbits. One of the most famous of these is the Watanabe heritable hyperlipidemic (WHHL) rabbit, which develops hypercholesterolemia and atherosclerosis spontaneously due to genetic and functional deficiencies of the low-density lipoprotein (LDL) receptor. The WHHL rabbit has been improved to develop myocardial infarction, and the new strain was designated the myocardial infarction-prone WHHL (WHHLMI) rabbit. This review summarizes the importance of selecting animal species for translational research in biomedical science, the development of WHHL and WHHLMI rabbits, their application to the development of hypocholesterolemic and/or antiatherosclerotic drugs, and future prospects regarding WHHL and WHHLMI rabbits.


Synaptic and intrinsic determinants of the phase resetting curve for weak coupling.
A phase resetting curve (PRC) keeps track of the extent to which a perturbation at a given phase advances or delays the next spike, and can be used to predict phase locking in networks of oscillators. The PRC can be estimated by convolving the waveform of the perturbation with the infinitesimal PRC (iPRC) under the assumption of weak coupling. The iPRC is often defined with respect to an infinitesimal current as z(i)(ϕ), where ϕ is phase, but can also be defined with respect to an infinitesimal conductance change as z(g)(ϕ). In this paper, we first show that the two approaches are equivalent. Coupling waveforms corresponding to synapses with different time courses sample z(g)(ϕ) in predictably different ways. We show that for oscillators with Type I excitability, an anomalous region in z(g)(ϕ) with opposite sign to that seen otherwise is often observed during an action potential. If the duration of the synaptic perturbation is such that it effectively samples this region, PRCs with both advances and delays can be observed despite Type I excitability. We also show that changing the duration of a perturbation so that it preferentially samples regions of stable or unstable slopes in z(g)(ϕ) can stabilize or destabilize synchrony in a network with the corresponding dynamics.


Magnetic resonance imaging in case of cortical apex syndrome caused by varicella zoster virus.
We present a case of orbital apex syndrome that developed after a re-activation of varicella zoster virus. The magnetic resonance imaging showed signs of a diffuse inflammation of the ipsilateral orbital cavity, external ocular muscles and optic nerve. Pattern visually evoked cortical potentials were non-recordable by stimulating the affected eye.


Storage in vivo of [3H]prostaglandins by rabbit blastocysts.
Day 6 rabbit blastocysts that had previously been incubated with 10 nM [3H]prostaglandin E2 (PGE2), [3H] PGF2 alpha, or 0.5 microCi [3H]water were surgically transferred to the uteri of day 6 pseudopregnant recipient rabbits. At 1, 3, and 20 h after transfer, blastocysts were collected from the recipient rabbits and evaluated for retention of [3H]ligands. The transferred day 6 blastocysts were able to retain a significant proportion of the initial PGs for up to 20 h in vivo. These PGs were not metabolized while they remained in the blastocysts in vivo, as assessed by HPLC. The transferred blastocysts appeared normal and viable after 20 h in vivo, since implantation sites could be detected visually. These data support our hypothesis that blastocysts can sequester PGs from their environment in vivo and retain them unmetabolized during the critical period just before implantation.


A methacrylic hyaluronic acid derivative for potential application in oral treatment of celiac disease.
Aim of this work was the synthesis of a methacrylic hyaluronic acid (HA) derivative and the production, via photocrosslinking, of related hydrogels loaded with an endopeptidase intended for a potential oral treatment of celiac disease. The methacrylic derivative of HA was prepared through a one-pot procedure involving the reaction with ethylenediamine (EDA) and methacrylic anhydride (MA). The obtained derivative, named HA-EDA-MA, was used to prepare photocrosslinked hydrogels loaded with a prolyl endopeptidase derived from Flavobacterium meningosepticum (PEP FM) able to detoxify gliadin. Obtained hydrogels were recovered as gels or freeze-dried powders. Hydrogels obtained as freeze-dried powders, are able to protect loaded enzyme from degradation due to freeze-drying process and from alteration during storage, overall in the presence of a cryoprotectant. All photocrosslinked HA-EDA-MA hydrogels (gels and powders) release PEP FM in simulated intestinal fluid in sustained manner and in active form. HA-EDA-MA hydrogels are nontoxic as demonstrated through in vitro studies on BALB 3T3 cells. Prepared hydrogels show a potential application for oral treatment of celiac disease thanks to the possibility to release enzymes able to detoxify the gliadin peptide that induces the immunogenic response.


Hypertensive update: highlights from the Sixth Report of the Joint National Committee on Prevention, Detection, Evaluation and Treatment of High Blood Pressure.
The National Heart, Lung and Blood Institute's Sixth Report of the Joint National Committee on Prevention, Detection, Evaluation and Treatment of High Blood Pressure (JNC VI) proposes a set of public health challenges to fight hypertension. The report proposes a system of risk stratification and pharmacologic therapy. Ongoing care of the person with high blood pressure (HBP) emphasizes the role of the primary care provider.


No evidence of differences in cognitive control in children with autism spectrum disorder or obsessive-compulsive disorder: An fMRI study.
Repetitive behaviors are among the core symptoms of both Autism Spectrum Disorder (ASD) and Obsessive-Compulsive Disorder (OCD) and are thought to be associated with impairments in cognitive control. However, it is still unknown how deficits in cognitive control and associated neural circuitry relate to the quality or severity of repetitive behavior in children with these disorders. Therefore, we investigated the behavioral and neural correlates of cognitive control using a modified stop-signal task in a multicenter study of children (aged 8-12 years) with ASD, OCD and typically developing (TD) children (N = 95). As both ASD and OCD have high levels of comorbidity with Attention Deficit/Hyperactivity Disorder (ADHD), we did an exploratory analysis addressing ADHD-symptoms. We found that children with ASD and OCD did not show deficits in cognitive control or changes in brain activity in task-relevant neural networks when compared to TD children. However, increased activity in prefrontal brain areas was associated with increased symptoms of comorbid ADHD. As such, this study does not support differences in cognitive control or associated neural circuitry in children with ASD and OCD, but rather suggests that changes in cognitive control in these disorders may be related to symptoms of comorbid ADHD.


Identifying key barriers in cationic polymer gene delivery to human T cells.
T cells have emerged as a therapeutically-relevant target for ex vivo gene delivery and editing. However, most commercially available reagents cannot transfect T cells and designing cationic polymers for non-viral gene delivery to T cells has resulted in moderate success. Here, we assess various barriers to successful gene transfer in the Jurkat human T cell line and primary human T cells. Using two polymers previously developed by our group, we show that uptake is one barrier to gene delivery in primary human T cells but is not predictive of successful gene delivery. We then probe intracellular pathways for barriers to gene transfer including endosomal acidification, autophagy, and immune sensing pathways. We find that endosomal acidification is slower and not as robust in human T cells compared to the model HeLa human cell line commonly used to evaluate cationic polymers for gene delivery. These studies inform the future design of cationic polymers for non-viral gene delivery to T cells, specifically, to rely on alternative endosomal release mechanisms rather than on pH-triggered release.


The yeast silent information regulator Sir4p anchors and partitions plasmids.
Circular plasmids containing telomeric TG1-3 arrays or the HMR E silencer segregate efficiently between dividing cells of the yeast Saccharomyces cerevisiae. Subtelomeric X repeats augment the TG1-3 partitioning activity by a process that requires the SIR2, SIR3, and SIR4 genes, which are also required for silencer-based partitioning. Here we show that targeting Sir4p to DNA directly via fusion to the bacterial repressor LexA confers efficient mitotic segregation to otherwise unstable plasmids. The Sir4p partitioning activity resides within a 300-amino-acid region (residues 950 to 1262) which precedes the coiled-coil dimerization motif at the extreme carboxy end of the protein. Using a topology-based assay, we demonstrate that the partitioning domain also retards the axial rotation of LexA operators in vivo. The anchoring and partitioning properties of LexA-Sir4p chimeras persist despite the loss of the endogenous SIR genes, indicating that these functions are intrinsic to Sir4p and not to a complex of Sir factors. In contrast, inactivation of the Sir4p-interacting protein Rap1p reduces partitioning by a LexA-Sir4p fusion. The data are consistent with a model in which the partitioning and anchoring domain of Sir4p (PAD4 domain) attaches to a nuclear component that divides symmetrically between cells at mitosis; DNA linked to Sir4p by LexA serves as a reporter of protein movement in these experiments. We infer that the segregation behavior of telomere- and silencer-based plasmids is, in part, a consequence of these Sir4p-mediated interactions. The assays presented herein illustrate two novel approaches to monitor the intracellular dynamics of nuclear proteins.


The interaction of melanoma cells with fibroblasts and endothelial cells in three-dimensional macromolecular matrices: a model for tumour cell invasion.
Comparative quantitative data are presented concerning the adhesion, proliferation and invasive behaviour of RPMI-3460 melanoma cells on (1) plain collagen gels, (2) monolayer cultures of fibroblasts and endothelial cells growing on the gel surface, and (3) the exposed endothelial and fibroblast extracellular matrices (ECMs). Both types of ECMs enhanced melanoma cell adhesion and proliferation (compared with plain gels) and had marked, but distinctive, effects on melanoma morphology. The thickness and composition of the ECMs was altered by treatment of the matrices with enzymes (trypsin, elastase and chondroitinase ABC) or by using ECMs produced by endothelial cells at various times after confluence. Variations in the thickness and composition of the ECMs had no effect on the behaviour of melanoma cells growing on these matrices; our results suggest that the glycoproteins and glycosaminoglycan ECM constituents removed by digestion with the enzymes do not play an important role in melanoma cell attachment, proliferation and migration. Melanoma cells plated on the surface of a plain collagen gel rapidly migrated down into the collagen matrix, with approximately 30% of the cells found within the gel after 6 days of incubation. Fibroblast and endothelial ECMs significantly and distinctively inhibited melanoma invasion into the underlying collagen gel. The extensive invasion of melanoma cells into the gel was not accompanied by hydrolysis of the collagen fibres. Conversely, fibroblast and endothelial ECMs, which acted as effective barriers, were extensively hydrolysed by the melanoma cells. The possible use of ECMs deposited on collagen in the study of melanoma local invasion (on fibroblast ECMs) and extravasation (on endothelial ECMs) is discussed.


Serum Müllerian inhibiting substance levels are lower in premenopausal women with breast precancer and cancer.
In preclinical studies, müllerian inhibiting substance (MIS) has a protective affect against breast cancer. Our objective was to determine whether serum MIS concentrations were associated with cancerous or precancerous lesions. Blood from 30 premenopausal women was collected and serum extracted prior to their undergoing breast biopsy to assess a suspicious lesion found on imaging or physical examination. Based on biopsy results, the serum specimens were grouped as cancer (invasive or ductal carcinoma in situ), precancer (atypical hyperplasia or lobular carcinoma in situ), or benign. Serum from women with cancer and precancer (p = .0009) had lower MIS levels than serum from women with benign disease. Our findings provide preliminary evidence for MIS being associated with current breast cancer risk, which should be validated in a larger population.


Hemorrhagic complications of severe pancreatitis.
Massive hemorrhage associated with pancreatitis is a rare but frequently lethal complication. Fifteen patients with this complication are presented. Bleeding occurred in four patients with necrotizing pancreatitis, in three patients with pancreatic abscesses, in seven patients with pseudocysts, and in one patient with chronic relapsing pancreatitis following longitudinal pancreaticojejunostomy. The initial presentation of hemorrhage was gastrointestinal in eight patients and retroperitoneal or intraperitoneal in seven. Abdominal pain with associated nausea and vomiting was present in all patients on admission. Duration of symptoms prior to hospitalization averaged 6 days. During hospitalization the 15 patients received a total of 512 units of blood for transfusions ranging from 8 to 177 units. Admission amylase values were of no benefit in assessing severity of the disease, but application of Ranson's criteria accurately predicted both severity and prognosis. The common denominator in all cases of bleeding appeared to be the presence of an overwhelming or continuing inflammatory process with necrosis and erosion of adjacent vascular and visceral structures. The overall mortality rate in the series was 53.3%. Those patients with hemorrhage associated with pseudocyst formation had the highest survival rates, whereas those with necrotizing pancreatitis and hemorrhage had an extremely poor response to aggressive medical and/or surgical management.


Genome-scale mutant fitness reveals versatile c-type cytochromes in Shewanella oneidensis MR-1.
Shewanella has been widely investigated for its metabolic versatility and use of a large number of extracellular electron acceptors. Many c-type cytochromes are responsible for this diversity, mainly in condition-specific fashions. By using genome-scale mutant fitness data, we studied which genes (particularly c-type cytochromes) were used to coordinate various electron transfer processes in the present work. First, by integrating fitness profiles with protein-protein interaction (PPI) networks, we showed that the genes with a high total fitness value were generally more important in PPI networks than those with low fitness values. Then, we identified genes that are important across many experiments, and further fitness analysis confirmed five versatile c-type cytochromes: ScyA (SO0264), PetC (SO0610), CcoP (SO2361), CcoO (SO2363) and CytcB (SO4666), which are considered to be crucial in most experimental conditions. Finally, we demonstrated a mediating role in the periplasm for the less-reported CytcB by combining protein structure, subcellular localization and disordered region analysis. Comparative genome analysis further revealed that it is distinctive in Shewanella species. Collectively, these results suggest that periplasmic electron transfer processes are more diverse and flexible than previously reported, giving insight for further experimental studies of Shewanella oneidensis MR-1.