Effects of various fragrant ingredients on desmopressin-induced fluid retention in mice.
Although fragrances are often used in aromatherapy for the treatment of edema, few studies on their diuretic and/or antiedematous activities have been performed. In this study, the effects of four types of fragrant ingredients (d-limonene, piperitone, alpha-pinene, and cinnamaldehyde) were examined in a mouse model of fluid retention. The mice were loaded with water after treatment with desmopressin (an antidiuretic hormone). In addition, zingerone, a pungent component of ginger which is considered to be effective in the treatment of edema, was examined. Moreover, their effects were compared with those of furosemide, a representative diuretic. Among the five types of fragrant ingredients examined, all except for cinnamaldehyde increased the urine volume in the fluid retention mouse model when administered at a dose of 100 mg/kg. In particular, d-limonene and zingerone significantly increased the urine volume. Thus the effects of these two ingredients were further examined at lower doses of 10 and 30 mg/kg. d-Limonene significantly increased the urine volume in a dose-dependent manner. Zingerone resulted in a significant increase in the urine volume only at a dose of 30 mg/kg. In normal mice, d-limonene did not affect the urine volume at the same doses. In contrast, zingerone significantly increased the urine volume in normal mice at a dose of 30 mg/kg. Furosemide significantly increased the urine volume in both the fluid-retentive and normal mice. These results indicate that both d-limonene and zingerone exhibit diuretic actions; however, the former fragrance only exerted an action in the fluid-retentive state. This different action suggests that d-limonene might be promising for the treatment of edema.


Decreased synthesis of acetylcholine accompanying impaired oxidation of pyruvic acid in rat brain minces.
The relation between pyruvate utilization and acetylcholine synthesis was investigated in minces of adult rat brain. The flux of pyruvate to acetylcholine was less than 1% of that to CO2; nevertheless, a number of agents which inhibited conversion of [1-14C]-pyruvate or [2-14C]pyruvate into 14CO2 were associated with corresponding decreases in the conversion of [2-14C]pyruvate into acetylcholine. The amount of acetylcholine produced by minces of whole rat brain, measured by g.l.c.-mass spectrometry, decreased similarly. Among the inhibitory compounds tested were 3-bromopyruvate, an irreversible inhibitor of pyruvate dehydrogenase; 2-oxobutyrate, a competitive inhibitor of pyruvate dehydrogenase; other 2-oxo acids; and amobarbital and pentobarbital. Linear-regression equations relating CO2 production to acetylcholine synthesis gave correlation coefficients of 0.89-0.93 for the combined observations. The inhibition of acetylcholine synthesis could not be attributed to inhibition of choline acetyltransferase. Incorporation of [2-14C]pyruvate into lipids, proteins and nucleic acids was effected less than that into acetylcholine. Under these experimental conditions, it was shown that pyruvate utilization can limit acetylcholine synthesis.


Characteristics of mothers who delivered the heaviest, average-weight, and lightest triplet sets.
We analyzed a cohort of 2850 live-born triplet sets to compare age, parity, stature, pre-gravid body mass index (BMI), and weekly weight gain in mothers who delivered triplets with a total weight in the 10th, 5th, and 1st deciles corresponding to the heaviest, average-weight, and lightest triplet sets, respectively. Mothers who delivered the heaviest triplets were significantly older, multiparous, taller, heavily built, and gained more weight compared with mothers of average-weight triplets. In contrast, except for higher parity, mothers of average-weight sets were not significantly different compared with mothers who delivered the lightest triplet sets. We concluded that parity was the only significant factor for increased total triplet weight in the first five deciles. However, the presence of other factors in addition to parity is needed for a triplet pregnancy to be included in the 10th decile.


Cognitive heterogeneity in Williams syndrome.
This study used the Woodcock-Johnson Tests of Cognitive Ability-Revised to investigate a wide range of cognitive abilities in people with Williams syndrome (WS). It involved a comparatively large sample of 31 people with WS, but took a case-series approach. The study addressed the widespread claims of a characteristic "WS cognitive profile" by looking for heterogeneity rather than homogeneity. People with WS showed a variety of preserved (significantly above mental age [MA]), expected (at MA), and significantly impaired (significantly below MA) levels of functioning. Such results provide clear evidence for heterogeneity in cognitive functions within WS. We found the most homogeneity on a test of phonological processing and a test of phonological short-term memory, with half of the WS sample performing at MA levels on these tests. Interestingly, no WS individual showed a weakness on a test of nonverbal reasoning, and only one WS individual showed a weakness on a test of verbal comprehension. In addition, we found that strengths on analysis-synthesis and verbal analogies occurred only for WS individuals with an MA less than 5.5 years (our sample median MA); people with an MA greater than 5.5 years performed at MA level on these 2 tests. Results also provided preliminary evidence for distinct subgroups of WS people based on their cognitive strengths and weaknesses on a broad range of cognitive functions. On the basis of the findings, caution should be made in declaring a single cognitive profile that is characteristic of all individuals with WS. Just as there is heterogeneity in genetic and physical anomalies within WS, not all WS individuals share the same cognitive strengths and weaknesses. Also, not all WS individuals show the profile of a strength in verbal abilities and a weakness in spatial functions.


Biotin-based strategies to explore the world of ubiquitin and ubiquitin-like modifiers.
A complex code of cellular signals is mediated by ubiquitin and ubiquitin-like (Ub/UbL) modifications on substrate proteins. The so-called Ubiquitin Code specifies protein fates, such as stability, subcellular localization, functional activation or suppression, and interactions. Hundreds of enzymes are involved in placing and removing Ub/UbL on thousands of substrates, while the consequences of modifications and the mechanisms of specificity are still poorly defined. Challenges include rapid and transient engagement of enzymes and Ub/UbL interactors, low stoichiometry of modified versus non-modified cellular substrates, and protease-mediated loss of Ub/UbL in lysates. To decipher this complexity and confront the challenges, many tools have been created to trap and identify substrates and interactors linked to Ub/UbL modification. This review focuses on an assortment of biotin-based tools we have developed for this purpose (for example BioUbLs, UbL-ID, BioE3), taking advantage of the strong affinity of biotin-streptavidin and the stringent lysis/washing approach allowed by it, paired with sensitive mass-spectrometry-based proteomic methods. Knowing how substrates change during development and disease, the consequences of substrate modification, and matching substrates to particular UbL-ligating enzymes will contribute new insights into how Ub/UbL signaling works and how it can be exploited for therapies.


In silico design of peptides with binding to the receptor binding domain (RBD) of the SARS-CoV-2 and their utility in bio-sensor development for SARS-CoV-2 detection.
The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has infected millions of people across the globe and created not only a health emergency but also a financial crisis. This virus attacks the angiotensin-converting enzyme 2 (ACE2) receptor situated on the surface of the host cell membrane. The spike protein of the virus binds to this receptor which is a critical step in infection. A molecule which can specifically stop this binding could be a potential therapeutic agent. In this study, we have tested 12 potential peptides which can bind to the receptor binding domain (RBD) of the spike protein of the virus and thus can potentially inhibit the binding of the latter on ACE2 receptors. These peptides are screened based on their binding with the RBD of the spike protein and aqueous stability, obtained using several atomistic molecular dynamic simulations. The potential of mean force calculation of peptides confirmed their binding to the RBD of the spike protein. Furthermore, two potential peptides were tested for use in a biosensing application for SARS-CoV-2 detection. Two types of biosensing platforms, a graphene sheet and a carbon nano tube (CNT) were tested. The peptides were modified in order to functionalize the graphene and CNT. Based on the interaction between the substrate, peptide and spike protein, the utility of the screened peptide for a given bio sensing platform is discussed and recommended.


Immobilization of wild ungulates: considerations and drugs.
Chemical immobilization of wild ungulates is a widely used tool of the wildlife researcher and is frequently necessary in the application of telemetry devices. Persons who immobilize wild animals simultaneously assume responsibility for the well-being of those animals. Considerations which must be made prior to immobilization, and some drugs used in ungulate immobilization are briefly discussed.


Functional outcome of infected endoprosthesis: A 20-year retrospective analysis.
Bone tumours are increasingly treated with limb-salvage surgeries. However, implant infection is a devastating complication, greatly affecting the functional outcome. Yet, data on functional outcome post-implant infection are scarce. This study aims to determine the functional outcome and implant survival of these patients. Patients' data on endoprosthetic replacement surgeries at our institution (January 1996-December 2016) was retrospectively reviewed. Information was available for 161 patients and was analysed using SPSS and SMART Partial Least Squares. Functional outcome was determined using the Musculoskeletal Tumor Society (MSTS) and Toronto Extremity Salvage Score (TESS) scoring system. Both mean rank MSTS (33.14 vs 87.02) and TESS (48.17 vs 85.13) scores were significantly lower in the infected group. These differences remained statistically significant after excluding amputation and rotationplasty cases within the infected group. Even after the resolution of infection, both MSTS and TESS remained significantly higher in the non-infected group. However, analysis of the infected group showed no significant differences in functional outcome between persistent and resolved infections (implant in-situ). Age significantly impacted the functional outcome for both the non-infected and infected groups, while local recurrence and metastasis significantly impacted the non-infected cases. Local tumour recurrence was lower in infected endoprosthetic patients (8.3% vs 10.5%). 56% of infected implants were removed; the majority were treated with two-stage revision surgery. Endoprosthesis infection worsens the overall functional outcome. Additional factors affecting functional outcome were age, presence of local recurrence and metastatic disease. Local tumour recurrence was lower amongst infected endoprosthesis cases, and >50% of infected implants were removed.


Correction of complex penile deformities by modified nesbit procedure asymmetric tunica albuginea excision.
Penile angulation combined with penile torsion is an uncommon finding in case of congenital or acquired penile curvature. We describe our technique for treatment of complex penile curvature by the excision of asymmetric ellipses of tunica albuginea. During the last 4 years, 4 patients with complex penile curvature underwent surgical correction by the excision of asymmetric ellipses of tunica albuginea. 3 patients had Peyronie's disease and 1 had congenital penile deviation. They ranged in age from 27 to 63 years. Convalescence was uneventful with good cosmetic correction of the penile torsion and lateral curvature in all patients. 1 patient reported shortening of the penis of about 2 cm, postoperatively. 2 patients reported decreased penile sensation, which resolved after 3 months. Erectile failure did not occur postoperatively in any case. During the last years many Authors have modified the original surgical procedure in an attempt to correct penile deformity. Unfortunately there are very few reports about correction of penile deformities combined with penile torsion. Our modified Nesbit surgical technique for complex deformities differs from the others with regard to the shape of the incision. In this manner, by shortening the longer corporeal body and creating a torque on the penis, it is possible to correct both problems: penile angulation and torsion. The technique is simple, bloodless, safe and reliable. In our experience good cosmetic and functional results have been achieved without complicated surgical interventions.


Increased number of FoxP3+CD4+ regulatory T cells in inflammatory bowel disease.
FoxP3 is a member of the forkhead/winged helix family of transcription factors and plays a critical role in the development and function of CD4+CD25+ regulatory T cells (Tregs). In this study, we performed an immunohistochemical evaluation of FoxP3-expressing cells in inflammatory bowel disease (IBD) mucosa. Mucosal FoxP3 expression was evaluated by immunohistochemistry in samples from normal (n=30), ulcerative colitis (UC) (n=53) and Crohn's disease (CD) (n=24) mucosa. There were no FoxP3-immunopositive cells in the normal colonic mucosa. In contrast, FoxP3-immunopositive cells were significantly increased in the inflamed regions of patients with active UC and CD. FoxP3-immunopositive cells completely coincided with some of the CD4-positive T cells. In conclusion, FoxP3-immunopositive Tregs were expanded in the inflamed mucosa of IBD patients. This suggests that these cells have impaired regulatory functions in the IBD mucosa.


Detection of time-varying signals in event-related fMRI designs.
In neuroimaging research on attention, cognitive control, decision-making, and other areas where response time (RT) is a critical variable, the temporal variability associated with the decision is often assumed to be inconsequential to the hemodynamic response (HDR) in rapid event-related designs. On this basis, the majority of published studies model brain activity lasting less than 4 s with brief impulses representing the onset of neural or cognitive events, which are then convolved with the hemodynamic impulse response function (HRF). However, electrophysiological studies have shown that decision-related neuronal activity is not instantaneous, but in fact, often lasts until the motor response. It is therefore possible that small differences in neural processing durations, similar to human RTs, will produce noticeable changes in the HDR, and therefore in the results of regression analyses. In this study we compare the effectiveness of traditional models that assume no temporal variance with a model that explicitly accounts for the duration of very brief epochs of neural activity. Using both simulations and fMRI data, we show that brief differences in duration are detectable, making it possible to dissociate the effects of stimulus intensity from stimulus duration, and that optimizing the model for the type of activity being detected improves the statistical power, consistency, and interpretability of results.


Can neurophysiologic measures serve as biomarkers for the efficacy of repetitive transcranial magnetic stimulation treatment of major depressive disorder?
Repetitive transcranial magnetic stimulation (rTMS) is an effective treatment for Major Depressive Disorder (MDD). There are clinical data that support the efficacy of many different approaches to rTMS treatment, and it remains unclear what combination of stimulation parameters is optimal to relieve depressive symptoms. Because of the costs and complexity of studies that would be necessary to explore and compare the large number of combinations of rTMS treatment parameters, it would be useful to establish reliable surrogate biomarkers of treatment efficacy that could be used to compare different approaches to treatment. This study reviews the evidence that neurophysiologic measures of cortical excitability could be used as biomarkers for screening different rTMS treatment paradigms. It examines evidence that: (1) changes in excitability are related to the mechanism of action of rTMS; (2) rTMS has consistent effects on measures of excitability that could constitute reliable biomarkers; and (3) changes in excitability are related to the outcomes of rTMS treatment of MDD. An increasing body of evidence indicates that these neurophysiologic measures have the potential to serve as reliable biomarkers for screening different approaches to rTMS treatment of MDD.


A novel autologous duraplasty in situ technique for the treatment of Chiari malformation Type I.
OBJECTIVE The purpose of this study was to introduce a novel autologous duraplasty procedure for the treatment of Chiari malformation Type I (CM-I). METHODS The authors retrospectively reviewed data from patients who had been diagnosed with CM-I and had undergone suboccipital decompression and autologous duraplasty in situ or synthetic dural graft duraplasty; patients were treated in the authors' department between 2011 and 2014. All procedures were performed by the same surgeon. The 2 duraplasty methods were compared in terms of surgical factors and complications. The authors assessed the neurological outcome and MRI-documented syrinx size at the 6-month follow-up visit. RESULTS Twenty-seven patients were enrolled in this study, 13 in the duraplasty in situ group and 14 in the synthetic dural graft duraplasty group. The results showed no significant differences between the duraplasty in situ and synthetic dural graft duraplasty groups in overall operative time (4.9 hours vs 4.1 hours; p = 0.070), estimated blood loss (229 ml vs 254 ml; p = 0.159), and duration of hospital stay after the operation (13.5 days vs 12.8 days; p = 0.808). In the duraplasty in situ group, 1 case of meningitis occurred (7.7%). In the synthetic dural graft duraplasty group, the complications included 1 case of meningitis (7.1%) and 1 CSF leak (7.1%). The mean cost of hospitalization in the duraplasty in situ group (CNY 23,354) was significantly lower than that in the synthetic dural graft duraplasty group (CNY 29,385; p = 0.036). CONCLUSIONS Compared with synthetic dural graft duraplasty, autologous duraplasty in situ is a safe, effective, and cost-effective procedure for the treatment of CM-I. The long-term outcome of this procedure requires investigation.


Salicylic Acid: a natural inducer of heat production in arum lilies.
For more than 50 years the identity of "calorigen," the agent that triggers pronounced heat production in the flowers and inflorescences of some thermogenic plants, remained obscure. Mass spectroscopic analysis of highly purified calorigen extracted from the male flowers of Sauromatum guttatum Schott (voodoo lily) revealed the presence of 2-hydroxybenzoic (salicylic) acid. Application of salicylic acid at 0.13 microgram per gram (fresh weight) to sections of the upper part of the plant's immature spadix, known as the appendix, led to temperature increases of as much as 12 Celsius degrees. These increases duplicated, in both magnitude and timing, the temperature increases produced by the crude calorigen extract. The sensitivity of appendix tissue to salicylic acid increases daily with the approach of anthesis and is controlled by the photoperiod. Thus, at least in some Arum lilies, salicylic acid functions as an endogenous regulator of heat production.


African Swine Fever Virus CD2v Protein Induces β-Interferon Expression and Apoptosis in Swine Peripheral Blood Mononuclear Cells.
African swine fever (ASF) is a hemorrhagic disease of swine characterized by massive lymphocyte depletion in lymphoid tissues due to the apoptosis of B and T cells, a process likely triggered by factors released or secreted by infected macrophages. ASFV CD2v (EP402R) has been implicated in viral virulence and immunomodulation in vitro; however, its actual function(s) remains unknown. We found that CD2v expression in swine PK15 cells induces NF-κB-dependent IFN-β and ISGs transcription and an antiviral state. Similar results were observed for CD2v protein treated swine PBMCs and macrophages, the major ASFV target cell. Notably, treatment of swine PBMCs and macrophages with CD2v protein induced apoptosis. Immunoprecipitation and colocalization studies revealed that CD2v interacts with CD58, the natural host CD2 ligand. Additionally, CD58 knockdown in cells or treatment of cells with an NF-κB inhibitor significantly reduced CD2v-mediated NF-κB activation and IFN-β induction. Further, antibodies directed against CD2v inhibited CD2v-induced NF-κB activation and IFN-β transcription in cells. Overall, results indicate that ASFV CD2v activates NF-κB, which induces IFN signaling and apoptosis in swine lymphocytes/macrophages. We propose that CD2v released from infected macrophages may be a significant factor in lymphocyte apoptosis observed in lymphoid tissue during ASFV infection in pigs.


Challenges in measuring obstetric fistula.
Obstetric fistula is gaining visibility within national and international public health agencies. With increased attention to this dire condition comes the demand for better epidemiologic data. This article assesses the current state of knowledge regarding population-based estimates of the incidence and prevalence of obstetric fistulas; proposes a method for better estimating these rates; and discusses the feasibility of this method. We show that there are no solid population-based estimates of the numbers of obstetric fistulas anywhere (the estimate from the most rigorous study being based on only 2 reported fistula cases); and for advocacy purposes, we recommend using the estimates from the 1990 Global Burden of Disease report. To estimate the incidence and prevalence of obstetric fistula, we propose an adaptation of the sibling-based method for direct estimation of maternal mortality. A series of questions are proposed for this use and sample size calculations are provided. The questions may require refinement, however, and we invite research groups to consider testing them.


Vegetarian diets in children and adolescents.
A well-balanced vegetarian diet can provide for the needs of children and adolescents. However, appropriate caloric intake should be ensured and growth monitored. Particular attention should be paid to adequate protein intake and sources of essential fatty acids, iron, zinc, calcium, and vitamins B₁₂ and D. Supplementation may be required in cases of strict vegetarian diets with no intake of any animal products. Pregnant and nursing mothers should also be appropriately advised to ensure that the nutritional needs of the fetus and infant are adequately met. Recommendations are provided. Adolescents on restricted vegetarian or other such diets should be screened for eating disorders.


The molecular genetics of keratin disorders.
Keratins are the type I and II intermediate filament proteins which form a cytoskeletal network within all epithelial cells. They are expressed in pairs in a tissue- and differentiation-specific fashion. Epidermolysis bullosa simplex (EBS) was the first human disorder to be associated with keratin mutations. The abnormal keratin filament aggregates observed in basal cell keratinocytes of some EBS patients are composed of keratins K5 and K14. Dominant mutations in the genes encoding these proteins were shown to disrupt the keratin filament cytoskeleton resulting in cells that are less resilient and blister with mild physical trauma. Identification of mutations in other keratin genes soon followed with attention focussed on disorders showing abnormal clumping of keratin filaments in specific cells. For example, in bullous congenital ichthyosiform erythroderma, clumping of filaments in the suprabasal cells led to the identification of mutations in the suprabasal keratins, K1 and K10. Mutations have now been identified in 18 keratins, all of which produce a fragile cell phenotype. These include ichthyosis bullosa of Siemens (K2e), epidermolytic palmoplantar keratoderma (K1, K9), pachyonychia congenita (K6a, K6b, K16, K17), white sponge nevus (K4, K13), Meesmann's corneal dystrophy (K3, K12), cryptogenic cirrhosis (K8, K18) and monilethrix (hHb6, hHb1).In general, these disorders are inherited as autosomal dominant traits and the mutations act in a dominant-negative manner. Therefore, treatment in the form of gene therapy is difficult, as the mutant gene needs to be inactivated. Ways of achieving this are actively being studied. Reliable mutation detection methods from genomic DNA are now available. This enables rapid screening of patients for keratin mutations. For some of the more severe phenotypes, prenatal diagnosis may be requested and this can now be performed from chorionic villus samples at an early stage of the pregnancy. This review article describes the discovery of, to date, mutations in 18 keratin genes associated with inherited human diseases.


A graph regularized non-negative matrix factorization method for identifying microRNA-disease associations.
MicroRNAs (miRNAs) play crucial roles in post-transcriptional regulations and various cellular processes. The identification of disease-related miRNAs provides great insights into the underlying pathogenesis of diseases at a system level. However, most existing computational approaches are biased towards known miRNA-disease associations, which is inappropriate for those new diseases or miRNAs without any known association information. In this study, we propose a new method with graph regularized non-negative matrix factorization in heterogeneous omics data, called GRNMF, to discover potential associations between miRNAs and diseases, especially for new diseases and miRNAs or those diseases and miRNAs with sparse known associations. First, we integrate the disease semantic information and miRNA functional information to estimate disease similarity and miRNA similarity, respectively. Considering that there is no available interaction observed for new diseases or miRNAs, a preprocessing step is developed to construct the interaction score profiles that will assist in prediction. Next, a graph regularized non-negative matrix factorization framework is utilized to simultaneously identify potential associations for all diseases. The results indicated that our proposed method can effectively prioritize disease-associated miRNAs with higher accuracy compared with other recent approaches. Moreover, case studies also demonstrated the effectiveness of GRNMF to infer unknown miRNA-disease associations for those novel diseases and miRNAs. The code of GRNMF is freely available at https://github.com/XIAO-HN/GRNMF/. Supplementary data are available at Bioinformatics online.


Metabolic mediators of the effects of body-mass index, overweight, and obesity on coronary heart disease and stroke: a pooled analysis of 97 prospective cohorts with 1·8 million participants.
Body-mass index (BMI) and diabetes have increased worldwide, whereas global average blood pressure and cholesterol have decreased or remained unchanged in the past three decades. We quantified how much of the effects of BMI on coronary heart disease and stroke are mediated through blood pressure, cholesterol, and glucose, and how much is independent of these factors. We pooled data from 97 prospective cohort studies that collectively enrolled 1·8 million participants between 1948 and 2005, and that included 57,161 coronary heart disease and 31,093 stroke events. For each cohort we excluded participants who were younger than 18 years, had a BMI of lower than 20 kg/m(2), or who had a history of coronary heart disease or stroke. We estimated the hazard ratio (HR) of BMI on coronary heart disease and stroke with and without adjustment for all possible combinations of blood pressure, cholesterol, and glucose. We pooled HRs with a random-effects model and calculated the attenuation of excess risk after adjustment for mediators. The HR for each 5 kg/m(2) higher BMI was 1·27 (95% CI 1·23-1·31) for coronary heart disease and 1·18 (1·14-1·22) for stroke after adjustment for confounders. Additional adjustment for the three metabolic risk factors reduced the HRs to 1·15 (1·12-1·18) for coronary heart disease and 1·04 (1·01-1·08) for stroke, suggesting that 46% (95% CI 42-50) of the excess risk of BMI for coronary heart disease and 76% (65-91) for stroke is mediated by these factors. Blood pressure was the most important mediator, accounting for 31% (28-35) of the excess risk for coronary heart disease and 65% (56-75) for stroke. The percentage excess risks mediated by these three mediators did not differ significantly between Asian and western cohorts (North America, western Europe, Australia, and New Zealand). Both overweight (BMI ≥25 to <30 kg/m(2)) and obesity (BMI ≥30 kg/m(2)) were associated with a significantly increased risk of coronary heart disease and stroke, compared with normal weight (BMI ≥20 to <25 kg/m(2)), with 50% (44-58) of the excess risk of overweight and 44% (41-48) of the excess risk of obesity for coronary heart disease mediated by the selected three mediators. The percentages for stroke were 98% (69-155) for overweight and 69% (64-77) for obesity. Interventions that reduce high blood pressure, cholesterol, and glucose might address about half of excess risk of coronary heart disease and three-quarters of excess risk of stroke associated with high BMI. Maintenance of optimum bodyweight is needed for the full benefits. US National Institute of Health, UK Medical Research Council, National Institute for Health Research Comprehensive Biomedical Research Centre at Imperial College Healthcare NHS Trust, Lown Scholars in Residence Program on cardiovascular disease prevention, and Harvard Global Health Institute Doctoral Research Grant.


Recombinants between bacteriophages T7 and T3 which productively infect F-plasmid-containing strains of Escherichia coli.
Recombinant phages between T7 and T3 have been isolated that grow well on strains of Escherichia coli that contain the F factor. One phage that has been characterized physically and genetically is predominately of the T7 genotype. Within this hybrid phage, two separate regions of T3 DNA have been located which are necessary for the phenotype of productive growth on F-containing strains. One of these, designated M1, contains the right part of gene 1 and continues through gene 1.3; the second, M2, appears to lie between gene 3 and gene 4.


Risk and protective factors associated with grandparent kinship caregivers' psychological distress in COVID-19: Kinship license status as a moderator.
COVID-19 and its related policy measures have increased the psychological distress of individuals, including grandparent kinship caregivers. Guided by the Resilience Model of Family Stress, Adjustment, and Adaptation, this study examines relationships between material hardship, parenting stress, social support, resilience and psychological distress of grandparent kinship caregivers during the COVID-19 pandemic, as well as the moderating role of kinship license status on these relationships. Kinship care licensing is a prerequisite to receiving financial assistance and other supporting services from the government. We administered a cross-sectional survey of grandparent kinship caregivers (N = 362) in the United States. Logistic regression results indicated that material hardship was associated with higher odds of experiencing psychological distress, whereas resilience and social support were associated with lower odds. Kinship license status moderated the relationships of social support and resilience with psychological distress. Results suggest that additional emergency funds and more tailored financial services should be provided to meet material needs, and interventions with a focus on resilience and social support are particularly needed. The moderating effects of license status indicate that some interventions should be specifically implemented among licensed kinship caregivers, whereas parallel services should be provided to kinship caregivers regardless of their license status.


Fast high-resolution brain metabolite mapping on a clinical 3T MRI by accelerated 1 H-FID-MRSI and low-rank constrained reconstruction.
Epitomizing the advantages of ultra short echo time and no chemical shift displacement error, high-resolution-free induction decay magnetic resonance spectroscopic imaging (FID-MRSI) sequences have proven to be highly effective in providing unbiased characterizations of metabolite distributions. However, its merits are often overshadowed in high-resolution settings by reduced signal-to-noise ratios resulting from the smaller voxel volumes procured by extensive phase encoding and the related acquisition times. To address these limitations, we here propose an acquisition and reconstruction scheme that offers both implicit dataset denoising and acquisition acceleration. Specifically, a slice selective high-resolution FID-MRSI sequence was implemented. Spectroscopic datasets were processed to remove fat contamination, and then reconstructed using a total generalized variation (TGV) regularized low-rank model. We further measured reconstruction performance for random undersampled data to assess feasibility of a compressed-sensing SENSE acceleration scheme. Performance of the lipid suppression was assessed using an ad hoc phantom, while that of the low-rank TGV reconstruction model was benchmarked using simulated MRSI data. To assess real-world performance, 2D FID-MRSI acquisitions of the brain in healthy volunteers were reconstructed using the proposed framework. Results from the phantom and simulated data demonstrate that skull lipid contamination is effectively removed and that data reconstruction quality is improved with the low-rank TGV model. Also, we demonstrated that the presented acquisition and reconstruction methods are compatible with a compressed-sensing SENSE acceleration scheme. An original reconstruction pipeline for 2D 1 H-FID-MRSI datasets was presented that places high-resolution metabolite mapping on 3T MR scanners within clinically feasible limits.


Role of vitamins B6, B12 and folic acid on hyperhomocysteinemia in a Pakistani population of patients with acute myocardial infarction.
Pakistani people belong to an ethnic group which has the highest rate of coronary artery disease (CAD). We investigated the possible correlation between deficiency of vitamins B6, B12 or folic acid and hyperhomocysteinemia in Pakistani patients with acute myocardial infarction (AMI). A case-control study was carried out involving 224 AMI patients (age 30-70 years; 55 females and 169 males) and 126 normal healthy subjects (age 31-70 years; 35 females and 91 males). Fasting venous blood was obtained from cases and controls. Serum was analyzed for folic acid and B12 using radioassays. Plasma was analyzed for pyridoxal phosphate (PLP; coenzymic form of B6) using a radioenzymatic assay and for total homocysteine using a fluorescence polarization immunoassay. Mean serum B12 concentration in AMI patients was found to be significantly lower than the mean for controls (241+/-185 pg/ml vs 608+/-341 pg/ml; p < 0.001). Mean serum folate level in patients was also found to be lower than controls (3.35+/-3.78 ng/ml vs 4.93+/-2.93 ng/ml), however, the differences were not statistically significant. Similarly, mean PLP concentration in plasma of cases (19.4+/-24.4 nmol/l) was lower than the concentration in controls (23.2+/-17.6 nmol/l), but the difference was not statistically significant. Mean plasma homocysteine level in AMI cases (18+/-8.36 micromol/l) was higher than the mean level in controls (16.4+/-4.9 micromol/l), but not to a significant extent. However, this mean homocysteine concentration in normal healthy subjects was among the highest reported in the literature and was significantly more than mean values reported in most Eastern and Western studies. Compared to controls, there was significantly greater deficiency of folate (32.5% vs 67.1%), B12 (3.2% vs 63.4%) and PLP (49.2% vs 74.1%) in AMI patients. Deficiencies of folate, B12 and PLP were defined as serum folate levels less than 3.5 ng/ml, serum levels of B12 less than 200 pg/ml and plasma PLP levels less than 20 nmol/l. Mean plasma homocysteine levels in smokers were found to be significantly higher in both cases and controls. Similarly, mean serum folate levels in smokers (compared to nonsmokers) were significantly lower in both cases and controls. Substantial nutritional deficiencies of these three vitamins along with mild hyperhomocysteinemia, perhaps through an interplay with the classical cardiovascular risk factors (highly prevalent in this population), could be further aggravating the risk of CAD in the Pakistani population.


Dopamine-dependent facilitation of LTP induction in hippocampal CA1 by exposure to spatial novelty.
In addition to its role in memory formation, the hippocampus may act as a novelty detector. Here we investigated whether attention to novel events can promote the associative synaptic plasticity mechanisms believed to be necessary for storing those events in memory. We therefore examined whether exposure to a novel spatial environment promoted the induction of activity-dependent persistent increases in glutamatergic transmission (long-term potentiation, LTP) at CA1 synapses in the rat hippocampus. We found that brief exposure to a novel environment lowered the threshold for the induction of LTP. This facilitatory effect was present for a short period following novelty exposure but was absent in animals that explored a familiar environment. Furthermore, the facilitation was dependent on activation of D1/D5 receptors. These findings support an important role for dopamine-regulated synaptic plasticity in the storage of unpredicted information in the CA1 area.


Identification of Coxiella burnetii type IV secretion substrates required for intracellular replication and Coxiella-containing vacuole formation.
Coxiella burnetii, the etiological agent of acute and chronic Q fever in humans, is a naturally intracellular pathogen that directs the formation of an acidic Coxiella-containing vacuole (CCV) derived from the host lysosomal network. Central to its pathogenesis is a specialized type IVB secretion system (T4SS) that delivers effectors essential for intracellular replication and CCV formation. Using a bioinformatics-guided approach, 234 T4SS candidate substrates were identified. Expression of each candidate as a TEM-1 β-lactamase fusion protein led to the identification of 53 substrates that were translocated in a Dot/Icm-dependent manner. Ectopic expression in HeLa cells revealed that these substrates trafficked to distinct subcellular sites, including the endoplasmic reticulum, mitochondrion, and nucleus. Expression in Saccharomyces cerevisiae identified several substrates that were capable of interfering with yeast growth, suggesting that these substrates target crucial host processes. To determine if any of these T4SS substrates are necessary for intracellular replication, we isolated 20 clonal T4SS substrate mutants using the Himar1 transposon and transposase. Among these, 10 mutants exhibited defects in intracellular growth and CCV formation in HeLa and J774A.1 cells but displayed normal growth in bacteriological medium. Collectively, these results indicate that C. burnetii encodes a large repertoire of T4SS substrates that play integral roles in host cell subversion and CCV formation and suggest less redundancy in effector function than has been found in the comparative Legionella Dot/Icm model.


[Krukenberg tumor--8 years after surgical treatment of gastric carcinoma].
The aim of the study was to discuss the case of Krukenberg tumor in a patient, eight years after the diagnosis and surgical treatment of gastric carcinoma. We have concluded that there was no safe period after which distant metastases may be excluded. In case of gastric carcinoma diagnosed in women, a regular gynecological examination is necessary due to a high propability of metastases into ovaries, even if the clinical stage of gastric carcinoma is low.


Investigation of epigenetics in kidney cell biology.
Epigenetics is the study of heritable changes in DNA or its associated proteins except mutations in gene sequence. Epigenetic regulation plays fundamental roles in the processes of kidney cell biology through the action of DNA methylation, chromatin modifications via epigenetic regulators and interaction via transcription factors, and noncoding RNA species. Kidney diseases, including acute kidney injury, chronic kidney disease, nephritic and nephrotic syndromes, pyelonephritis and polycystic kidney diseases are driven by aberrant activity in numerous signaling pathways in even individual kidney cell. Epigenetic alterations, including DNA methylation, histone acetylation and methylation, noncoding RNAs, and protein posttranslational modifications, could disrupt essential pathways that protect the renal cells from uncontrolled growth, apoptosis and establishment of other renal associated syndromes, which have been recognized as one of the critical mechanisms for regulating functional changes that drive and maintain the kidney disease phenotype. In this chapter, we briefly summarize the epigenetic mechanisms in kidney cell biology and epigenetic basis of kidney development, and introduce epigenetic techniques that can be used in investigating the molecular mechanism of kidney cell biology and kidneys diseases, primarily focusing on the integration of DNA methylation and chromatin immunoprecipitation technologies into kidney disease associated studies. Future studies using these emerging technologies will elucidate how alterations in the renal cell epigenome cooperate with genetic aberrations for kidney disease initiation and progression. Incorporating epigenomic testing into the clinical research is essential to future studies with epigenetics biomarkers and precision medicine using emerging epigenetic therapies.


Morphology and peroxidase cytochemistry of mouse promonocytes, monocytes, and macrophages.
Mouse promonocytes have been identified and studied in cultures of bone marrow cells. These cells have a diameter of 14-20 micro, and in stained preparations reveal a large, indented or folded nucleus, and basophilic, finely granular cytoplasm. The living promonocyte viewed by phase contrast shows additional features: nucleoli, small dense bodies, and vesicles in the cytoplasm adjacent to the nuclear hilus, and slight membrane ruffling. Prominent ultrastructural components of promonocytes include a well developed Golgi apparatus, small numbers of centrosomal granules and vacuoles, extensive ribosomal aggregates, and finger-like projections of the cell surface. Promonocytes engage in pinocytosis and phagocytosis, but they are less active in these functions than are peripheral blood monocytes of peritoneal macrophages. Promonocytes are positive for peroxidase, the reaction product being localized to granules most of which are centrally situated in the cell. Monocytes in blood or in inflammatory peritoneal exudates display much smaller numbers of peroxidase-positive granules, and various types of mature mouse macrophages are peroxidase negative.


Technical nuances to minimize common complications of deep brain stimulation.
The implantation of deep brain stimulator electrodes is associated with infrequent complications. These complications are consistent across prospective trials and include infection, skin erosion, hemorrhage, and lead misplacement. Nuances of surgical technique can be used to minimize the risk of these commonly noted complications. Several of these technical nuances are highlighted in this video submission. The video can be found here: https://youtu.be/GL09W9p013g .


Risk of Stroke and Myocardial Infarction After Herpes Zoster in Older Adults in a US Community Population.
To assess the risk of stroke and myocardial infarction (MI) after herpes zoster in a US community population of older adults. We performed a community cohort study (January 1, 1986, to October 1, 2011) comparing the risk of stroke and MI in 4862 adult residents of Olmsted County, Minnesota, 50 years and older with and without herpes zoster and 19,433 sex- and age-matched individuals with no history of herpes zoster. Odds ratios are presented for MI and stroke at 3, 6, 12, and 36 months after index herpes zoster plus hazard ratios for long-term risk (up to 28.6 years). Individuals with herpes zoster had more risk or confounding factors for MI and stroke, suggesting that they had worse health status overall. When controlling for the multiple risk factors, those with herpes zoster were at increased risk for stroke at 3 months after herpes zoster compared with those without a history of herpes zoster (odds ratio, 1.53; 95% CI, 1.10-2.33; P=.04). The association between herpes zoster and MI at 3 months was not robust across analytic methods. Herpes zoster was not associated with an increased risk of stroke or MI at any point beyond 3 months. Herpes zoster was associated with only a short-term increased risk of stroke, which may be preventable with the prevention of herpes zoster.


Dichloroacetate--its in vivo effects on carbohydrate metabolism in the conscious dog.
The effects of sodium dichloroacetate (DCA) on carbohydrate metabolism in conscious, 48-h-fasted dogs were examined using the hepatic A-V difference technique and a double isotope infusion technique (3H-glucose to measure glucose production and 14C-alanine to assess gluconeogenesis). DCA infusion (0.4 mg/kg-min) resulted in an 82 +/- 1% fall in the arterial plasma alanine level and a 53 +/- 8% fall in the arterial whole blood lactate level. Hepatic uptake of alanine and lactate fell 67 +/- 5% and 59 +/- 15%, respectively, although the fractional extraction of these intermediates was not altered. DCA decreased the conversion of circulating alanine and lactate to glucose but by only 41 +/- 7%, suggesting that a slight increase in the efficiency of the intrahepatic gluconeogenic process took place. This may be explained by the decrease in the plasma insulin level (39 +/- 9%) that occurred in the presence of an unchanged plasma glucagon concentration. Despite the substantial fall in the levels of gluconeogenic precursors in blood and the considerable decrease in their rate of conversion to glucose, the overall rates of glucose production and the blood glucose concentration were not altered by DCA. These data indicate that the alanine and lactate supplied by the periphery after a 48 h fast in the dog are not essential for the acute maintenance of glucose production or euglycemia. They suggest, further, that a compensatory increase in glucose production can occur by drawing on an alternate intrahepatic carbon source, the nature of which and signal for which remain unclear.


Form and function of the bulbus arteriosus in yellowfin tuna (Thunnus albacares), bigeye tuna (Thunnus obesus) and blue marlin (Makaira nigricans): static properties.
The juxtaposition of heart and gills in teleost fish means that the Windkessel function characteristic of the whole mammalian arterial tree has to be subserved by the extremely short ventral aorta and bulbus arteriosus. Over the functional pressure range, arteries from blue marlin (Makaira nigricans) and yellowfin tuna (Thunnus albacares) have J-shaped pressure-volume (P-V) loops, while bulbi from the same species have r-shaped P-V loops, with a steep initial rise followed by a compliant plateau phase. The steep initial rise in pressure is due to the geometry of the lumen. The interactions between radius, pressure and tension require a large initial pressure to open the bulbar lumen for flow. The plateau is due to the unique organization of the bulbar wall. The large elastin:collagen ratio, limited amount of collagen arranged circumferentially, lack of elastin lamellae and low hydrophobicity of the elastin itself all combine to lower stiffness, increase extensibility and allow efficient recoil. Even though the modulus of bulbus material is much lower than that of an artery, at large volumes the overall stiffness of the bulbus increases rapidly. The morphological features that give rise to the special inflation characteristics of the bulbus help to extend flow and maintain pressure during diastole.


Merohedral icosahedral M48 (M = CoII, NiII) cage clusters supported by thiacalix[4]arene.
Cage clusters are a discrete chemically and topologically diverse family of molecule-based functional materials. Presented here are two isostructural M48 (M = CoII for LSHU01, NiII for LSHU02) cage clusters with a merohedral icosahedral cage structure featuring 12 M4-TC4A (H4TC4A, p-tert-butylthiacalix[4]arene) second building units as vertices and 18 asymmetric 5-(1H-tetrazol-1-yl)isophthalate ligands as faces. They are the highest-nuclearity cage compounds of CoII and NiII. The activated Co48 cage exhibited high selectivity in the sorption of C3H8 over CH4 under ambient conditions. Frequency response experiments indicated that the extrinsic voids and matrix interface of the activated crystalline samples are primarily responsible for the observed gas adsorption performance.


The intra-lesional variation of type, level of invasion, and tumour thickness of primary cutaneous malignant melanoma.
In order to evaluate the prognostic importance of type of melanoma, level of invasion and tumour thickness in a retrospective study of cutaneous melanomas, it is prerequisite that the available histological material be representative. To investigate this condition 55 primary cutaneous melanomas were studied randomly by serial block technique. The histological classification of each tissue block as to 1) type of melanoma, 2) level of invasion, and 3) tumour thickness was correlated with the overall classification of the corresponding whole tumour. The typing included 6 groups: Superficial spreading melanoma, lentigo maligna melanoma, nodular melanoma, acral lentiginous melanoma, melanoma of unclassifiable type, and nevoid lesion. The depth of tumour invasion was graded by 5 levels according to Clark et al. The tumour thickness was measured in mm as described by Breslow, and arranged in 6 groups (0--0.75 mm, 0.76--1.50 mm, 1.51-2.25 mm, 2.26--3.00 mm. 3.01--6.00 mm, and more than 6.00 mm). The study revealed a consistently of 95% (52/55) between the type of the central cross section and the type of the whole tumour. In 96% (53/55) of the central cross sections the level of invasion was equal to that of the whole tumour. In 95% (52/55) of the central cross sections the tumour thickness was situated in the same group as that of the whole tumour. It is concluded that type, level and thickness of malignant melanomas can be evaluated in retrospective studies if central cross sections of the melanomas are available for histological examination.


Pearls, perils, and pitfalls in the use of the electroencephalogram.
Despite advances in neuroimaging techniques over the past three decades that have helped in identifying structural lesions of the central nervous system, electroencephalography (EEG) continues to provide valuable insight into brain function by demonstrating focal or diffuse background abnormalities and epileptiform abnormalities. It is an extremely valuable test in patients suspected of epilepsy and in patients with altered mental status and coma. Patterns in the EEG make it possible to clarify the seizure type; it is indispensable for the diagnosis of nonconvulsive status epilepticus and for separating epileptic from other paroxysmal (nonepileptic) episodes. There are EEG patterns predictive of the cause of the encephalopathy (i.e., triphasic waves in metabolic encephalopathy) or the location of the lesion (i.e., focal polymorphic delta activity in lesions of the subcortical white matter). The various EEG characteristics of infantile, childhood, and adult epilepsies are described as well as the EEG patterns that are morphologically similar to interictal/ictal epileptiform discharges but unrelated to epilepsy. An EEG is most helpful in determining the severity and, hence, the prognosis of cerebral dysfunction. Lastly, EEG is extremely helpful in assessing normal or abnormal brain functioning in a newborn because of the serious limitation in performing an adequate neurologic examination on the neonate who is intubated or paralyzed for ventilatory control. Under such circumstances, the EEG may be the only available tool to detect an encephalopathic process or the occurrence of epileptic seizures.


A missense variant in SHARPIN mediates Alzheimer's disease-specific brain damages.
Established genetic risk factors for Alzheimer's disease (AD) account for only a portion of AD heritability. The aim of this study was to identify novel associations between genetic variants and AD-specific brain atrophy. We conducted genome-wide association studies for brain magnetic resonance imaging measures of hippocampal volume and entorhinal cortical thickness in 2643 Koreans meeting the clinical criteria for AD (n = 209), mild cognitive impairment (n = 1449) or normal cognition (n = 985). A missense variant, rs77359862 (R274W), in the SHANK-associated RH Domain Interactor (SHARPIN) gene was associated with entorhinal cortical thickness (p = 5.0 × 10-9) and hippocampal volume (p = 5.1 × 10-12). It revealed an increased risk of developing AD in the mediation analyses. This variant was also associated with amyloid-β accumulation (p = 0.03) and measures of memory (p = 1.0 × 10-4) and executive function (p = 0.04). We also found significant association of other SHARPIN variants with hippocampal volume in the Alzheimer's Disease Neuroimaging Initiative (rs3417062, p = 4.1 × 10-6) and AddNeuroMed (rs138412600, p = 5.9 × 10-5) cohorts. Further, molecular dynamics simulations and co-immunoprecipitation indicated that the variant significantly reduced the binding of linear ubiquitination assembly complex proteins, SHPARIN and HOIL-1 Interacting Protein (HOIP), altering the downstream NF-κB signaling pathway. These findings suggest that SHARPIN plays an important role in the pathogenesis of AD.


Id1 is a common downstream target of oncogenic tyrosine kinases in leukemic cells.
Oncogenic tyrosine kinases, such as BCR-ABL, TEL-ABL, TEL-PDGFbetaR, and FLT3-ITD, play a major role in the development of hematopoietic malignancy. They activate many of the same signal transduction pathways. To identify the critical target genes required for transformation in hematopoietic cells, we used a comparative gene expression strategy in which selective small molecules were applied to 32Dcl3 cells that had been transformed to factor-independent growth by these respective oncogenic alleles. We identified inhibitor of DNA binding 1 (Id1), a gene involved in development, cell cycle, and tumorigenesis, as a common target of these oncogenic kinases. These findings were prospectively confirmed in cell lines and primary bone marrow cells engineered to express the respective tyrosine kinase alleles and were also confirmed in vivo in murine models of disease. Moreover, human AML cell lines Molm-14 and K562, which express the FLT3-ITD and BCR-ABL tyrosine kinases, respectively, showed high levels of Id1 expression. Antisense and siRNA based knockdown of Id1-inhibited growth of these cells associated with increased p27(Kip1) expression and increased sensitivity to Trail-induced apoptosis. These findings indicate that Id1 is an important target of constitutively activated tyrosine kinases and may be a therapeutic target for leukemias associated with oncogenic tyrosine kinases.


Beneficial Microbes: The pharmacy in the gut.
The scientific evidence supporting the gut microbiome in relation to health maintenance and links with various disease states afflicting humans, from metabolic to mental health, has grown dramatically in the last few years. Strategies addressing the positive modulation of microbiome functionality associated with these disorders offer huge potential to the food and pharmaceutical industries to innovate and provide therapeutic solutions to many of the health issues affecting modern society. Such strategies may involve the use of probiotics and prebiotics as nutritional adjunct therapies. Probiotics are generally recognized to be a good form of therapy to keep harmful, intestinal microorganisms in check, aid digestion and nutrient absorption, and contribute to immune function. Probiotics are reported to improve microbial balance in the intestinal tract and promote the return to a baseline microbial community following a perturbing event (dysbiosis) such as antibiotic therapy. Prebiotics are selectively fermented ingredients that allow specific changes, both in the composition and/or activity in the gastrointestinal microflora, which confers benefits upon host well-being and health.


Routine immunization practices: use of topical anesthetics and oral analgesics.
Immunization pain is a global public health issue. Despite an abundance of data that demonstrate the efficacy of local anesthetics for decreasing immunization pain, their adoption in practice has not been determined. Our objective was to evaluate analgesic use during childhood immunization. We used a cluster-sampling survey of pediatricians in the greater Toronto area (who administer immunizations) and multiparous women. By using a self-administered survey, pediatricians reported frequency of analgesic use in their practice for 2 phases of immunization: injection (needle puncture and vaccine administration) and postinjection (hours to days postvaccination). By using an interviewer-administered face-to-face survey, mothers reported analgesic practices for their children. Of 195 eligible pediatricians, 140 (72%) responded. During the injection phase, 58% rarely or never used analgesics compared with 11% for the postinjection phase. During injection, the local anesthetics lidocaine-prilocaine and tetracaine were used at least sometimes in 12% and 2% of the practices, respectively, whereas acetaminophen and ibuprofen were used in 81% and 46%, respectively. Postinjection, acetaminophen and ibuprofen were used in 89% and 56% of practices. Of 257 eligible mothers, 200 (78%) participated. During injection, analgesics were used in 25% of immunizations (acetaminophen [87%], ibuprofen [7%], and lidocaine-prilocaine [6%]). Postinjection, analgesics were used in 33% of immunizations (acetaminophen [86%] and ibuprofen [14%]). A minority of pediatricians and mothers use topical local anesthetics during childhood immunization despite evidence to support their use. Oral analgesics are used more commonly, but this practice is not consistent with scientific evidence. Knowledge-translation strategies are needed to increase the use of local anesthesia.


[History of solving medical problems of safety provisions of altitude flights].
The paper outlines the basic landmarks of the activity of the workers of the RF State Research Test Institute of Aviation and Space Medicine who have solved the problems of altitude flight safety provision, designed various sets of oxygen and altitude equipment. The main results of the investigations made by the Altitude Department of the Institute are summed up and some promising directions of its workers' activities are shown.


Drinking to our health: can beverage companies cut calories while maintaining profits?
Carbonated soft drinks and other beverages make up an increasing percentage of energy intake, and there are rising public health concerns about the links between consumption of sugar-sweetened beverages and weight gain, obesity, and other cardiometabolic problems. In response, the food and beverage industry claims to be reformulating products, reducing package or portion sizes and introducing healthier options. Comparative analysis on various changes and their potential effects on public health are needed. We conduct a case study using the two largest and most influential producers of sweetened beverages, The Coca-Cola Company and PepsiCo Inc., who together control 34% of the global soft drink market, examining their product portfolios globally and in three critical markets (the United States, Brazil and China) from 2000 to 2010. On a global basis, total revenues and energy per capita sold increased, yet the average energy density (kJ 100 mL(-1) ) sold declined slightly, suggesting a shift to lower-calorie products. In the United States, both total energy per capita and average energy density of beverages sold decreased, while the opposite was true in the developing markets of Brazil and China, with total per capita energy increasing greatly in China and, to a lesser extent, in Brazil.


Highlights for the management of a child with proteinuria and hematuria.
The identification of hematuria or proteinuria in an otherwise healthy child can cause anxiety to both the family and the pediatrician. The etiology of hematuria and proteinuria includes a long list of conditions, and detailed workup can be exhaustive, expensive and not essential in most of the patients. As will be described in this paper, most of the children with proteinuria or hematuria have a benign etiology. The primary role of the pediatrician is to identify hematuria/proteinuria, recognize the common causes of hematuria/proteinuria, and more importantly identify children with serious conditions that need referral to the nephrologist in a timely manner.


Infections in Pediatric Burn Patients: An Analysis of One Hundred Eighty-One Patients.
Background: Infectious complications are one of the most life-threatening complications and result in substantial mortality and morbidity in children who have been burned. The goal of the study is to assess the risk factors for sepsis in pediatric burn patients in a referral hospital. Methods: This study was performed at the Pediatric Burn Unit of Ankara Child Health and Diseases Hematology Oncology Training and Research Hospital during the period between January 2014 and June 2017. The patients were evaluated for age, sex, burn etiology, burned body surface area (BSA), the presence of inhalation injury, sepsis, positive cultures, the micro-organisms cultured samples, and septic focus. Results: A total of 181 patients were included in the study. The most common cause of burns was scalds in 120 patients (66.3%). Forty-one patients (22.7%) developed health-care-associated infection and sepsis. Gram-negative micro-organisms were isolated in 40 (97.6%) patients (Acinetobacter spp., Pseudomonas aeruginosa, Klebsiella pneumonia) with sepsis. Carbapenem resistance was detected in 31 (93.8%) of 40 patients. Mortality was observed in 11 patients (6.1%) in the group with sepsis. Burn surface area, burn depth, C-reactive protein (CRP) values, mortality, Garcés index, and Baux index were higher in the group with sepsis (p < 0.05). Multiple regression analysis revealed that mechanism of injury (flame), burned BSA ≥25%, C-reactive protein ≥6 mg/dL (area under the curve [AUC]: 0.76 p < 0.001 and AUC: 0.90, p < 0.001, respectively) at admission were independent parameters for development of sepsis in pediatric burn patients. Conclusion: Multi-drug-resistant Pseudomonas aeruginosa and Acinetobacter baumannii were important agents of blood stream infection in burned children. Burned BSA ≥25% and CRP ≥6 mg/dL were risk factors for developing sepsis in pediatric burn patients.


Preparedness and response of assisted living facilities toward COVID-19 infection control in Houston, Texas, USA.
Long-term care facilities (LTCFs) including assisted living facilities (ALFs) are hubs for high transmission and poor prognosis of COVID-19 among the residents who are more susceptible due to old age and comorbidities. Houston Health Department conducted assessments of ALFs within the City of Houston to determine preparedness and existing preventive measures at the facilities. Onsite assessments were conducted at ALFs using a modified CDC Infection Control Assessment and Response (ICAR) Tool. Data was obtained on IPC measures, training, testing, vaccination etc. Data was analyzed, frequencies generated, and bivariate associations determined. A total of 118 facilities were assessed and categorized into small scale 46 (39%), medium scale 47 (40%), and large scale 25 (21%). The facilities had 2431 residents and 2290 staff. Thirty-one (26%) facilities reported an outbreak in 2020, while 14 (12%) had an ongoing outbreak. Twenty-three (97%) large-scale and 12 (26%) small-scale facilities had COVID-19 testing program. Vaccination coverage among residents ranged from 99% in large-scale to 40% in small-scale facilities but was smaller among staff at 748 (45%) in large scale, 71 (36%) in small scale, and 193 (45%) in medium scale. While 24 (96%) large-scale and 34 (77%) of small-scale facilities conducted staff training staff on IPC practices, 22 (92%) of large-scale and 19 (56%) of small-scale facility staff demonstrated capacity (p = 0.01), respectively. Visitor screening was done at 100% of large-scale and 80% of small-scale and the medium-scale ALFs. Assisted living facilities within the city of Houston are at various levels of preparedness and interventions with respect to COVID-19 response.


Hiring Help for the Home: Household Services in the Twentieth Century.
[outsourcinghousehold laborcommodificationservicespaid labor ]] The division between home and market has long been a key dimension of family life and shifts across the home/market boundary are important for both gender inequality and the family. Two shifts across this boundary occurred in the twentieth century: women's movement into paid labor and a decline in household labor. One stylized conception suggests that as women moved into paid labor, they used their new resources to purchase replacements. This article takes up this question, asking whether women's movement into market labor led to the commodification of the home. It does so by combining evidence from primary and secondary sources about women's reliance on services to offer a picture of how services have been used. The article argues that household labor is infused with emotional and relational content which has made it difficult to replace.


Chemokines have diverse abilities to form solid phase gradients.
Chemokines play critical roles in leukocyte recruitment into sites of inflammation such as rheumatoid arthritis (RA). While chemokines immobilized on endothelium (solid-phase), but not soluble chemokines, direct rolling leukocytes to firmly adhere to endothelium, soluble and solid-phase chemokine gradients may play important roles in leukocyte extravasation into the tissue. In this study, we have sought to determine (1) if chemokines can be immobilized on structures in the extravascular space, (2) the mechanisms by which chemokines may be immobilized, and (3) if different chemokines have similar potentials to form solid-phase gradients. While secreted alkaline phosphatase (SEAP)-tagged chemokines SLC (CCL21), TARC (CCL17), and RANTES (CCL5) bound to mast cells and the extracellular matrix (ECM) in RA synovium under physiologic salt conditions, MCP1 (CCL2), MIP1 alpha (CCL3), MIP1 beta (CCL4), and fractalkine (FKN, CX3CL1) fusion proteins did not detectably bind. Chemokine binding to ECM and mast cells in situ and to immobilized heparin was inhibited by high salt and glycosaminoglycans (GAGs) heparin, heparan sulfate, chondroitin sulfate, and dermatan sulfate, but not by dextran or hyaluronan, indicating that the chemokines bind to highly sulfated GAGs. Chemokine binding to synovial structures correlated strongly with avidity of chemokine binding to heparin (SLC > TARC > RANTES > MIP1 beta > MCP1 > MIP1 alpha > FKN). A RANTES mutant with decreased avidity for heparin was not able to bind to ECM or mast cells. Thus, these data indicate that chemokines can bind to ECM and mast cell granule constituents in situ via interactions with GAGs. Further, only a subset of chemokines were able to bind efficiently to structures in the extravascular space, indicating that chemokines may form different types of gradients based on their GAG binding ability and that chemotactic gradients in tissues may be quite complex.


Angiotensin II, type 2 receptor is not involved in the angiotensin II-mediated pro-atherogenic process in ApoE-/- mice.
Angiotensin II (Ang II) accelerates atherogenesis in ApoE mice via the angiotensin II, type 1 receptor (AT1) while the type 2 receptor (AT2) is suggested to counteract atherogenesis. To confirm and further explore this possibility, we studied the effect of AT2 receptor antagonism on Ang II-accelerated atherosclerosis. ApoE mice were fed a standard or high cholesterol diet (1.25%) for 4 weeks. Mice on each diet were treated with either Ang II (0.5 microg/kg per min) or Ang II in combination with PD123319 (3 mg/kg per day). Plaque distribution was assessed by en face quantification of the thoracic aorta and in cross-sections of the aortic root. Mean arterial pressure (MAP) was measured. AT1 and AT2 receptor expression were analysed using real-time polymerase chain reaction (PCR) and the localization of the AT2 receptor protein confirmed with immunohistochemistry. Ang II infusion increased MAP only in mice on a standard diet (P < 0.001). Regardless of diet, Ang II-infused mice had 22-30 times increased plaque area in the thoracic aorta (P < 0.001 for both). Ang II had no effect on plaque in the aortic root. Plaque area was not affected by PD123319. AT2 receptor was heavily expressed in the plaques and increased six- to ninefold by a high cholesterol diet and Ang II infusion (P < 0.01). Ang II increases the extent of atherosclerosis in ApoE mice. Despite up-regulation of the AT2 receptor, we found no support for an effect of the AT2 receptor on atherogenesis in this model.


Interspecies variation in survival and growth of filamentous heterotrophic bacteria in response to UVC radiation.
Ultraviolet radiation is an important environmental constraint on the evolution of life. In addition to its harmful effects, ultraviolet radiation plays an important role in generating genetic polymorphisms and acting as a selective agent. Understanding how prokaryotes cope with high radiation can give insights on the evolution of life on Earth. Four representative filamentous bacteria from the family Cytophagaceae with different pigmentation were selected and exposed to different doses of UVC radiation (15-32,400Jm(-2)). The effect of UVC radiation on bacterial survival, growth and morphology were investigated. Results showed high survival in response to UVC for Rudanella lutea and Fibrisoma limi, whereas low survival was observed for Fibrella aestuarina and Spirosoma linguale. S. linguale showed slow growth recovery after ultraviolet exposure, R. lutea and F. limi showed intermediate growth recovery, while F. aestuarina had the fastest recovery among the four tested bacteria. In terms of survival, S. linguale was the most sensitive bacterium whereas R. lutea and F. limi were better at coping with UVC stress. The latter two resumed growth even after 2h exposure (∼10,800Jm(-2)). Additionally, the ability to form multicellular filaments after exposure was tested using two bacteria: one representative of the high (R. lutea) and one of the low (F. aestuarina) survival rates. The ability to elongate filaments due to cell division was preserved but modified. In R. lutea 10min exposure reduced the average filament length. The opposite was observed in F. aestuarina, where the 5 and 10min exposures increased the average filament length. R. lutea and F. limi are potential candidates for further research into survival and resistance to ultraviolet radiation stress.


Genetic diversity of the KIR/HLA system and susceptibility to hepatitis C virus-related diseases.
The variability in the association of host innate immune response to Hepatitis C virus (HCV) infection requires ruling out the possible role of host KIR and HLA genotypes in HCV-related disorders: therefore, we therefore explored the relationships between KIR/HLA genotypes and chronic HCV infection (CHC) as they relate to the risk of HCV-related hepatocarcinoma (HCC) or lymphoproliferative disease progression. We analyzed data from 396 HCV-positive patients with CHC (n = 125), HCC (118), and lymphoproliferative diseases (153), and 501 HCV-negative patients. All were HIV and HBV negative. KIR-SSO was used to determine the KIR typing. KIR2DL5 and KIR2DS4 variants were performed using PCR and GeneScan analysis. HLA/class-I genotyping was performed using PCR-sequence-based typing. The interaction between the KIR gene and ligand HLA molecules was investigated. Differences in frequencies were estimated using Fisher's exact test, and Cochran-Armitage trend test. The non-random association of KIR alleles was estimated using the linkage disequilibrium test. We found an association of KIR2DS2/KIR2DL2 genes, with the HCV-related lymphoproliferative disorders. Furthermore, individuals with a HLA-Bw6 KIR3DL1+ combination of genes showed higher risk of developing lymphoma than cryoglobulinemia. KIR2DS3 gene was found to be the principal gene associated with chronic HCV infection, while a reduction of HLA-Bw4 + KIR3DS1+ was associated with an increased risk of developing HCC. Our data highlight a role of the innate-system in developing HCV-related disorders and specifically KIR2DS3 and KIR2D genes demonstrated an ability to direct HCV disease progression, and mainly towards lymphoproliferative disorders. Moreover the determination of KIR3D/HLA combination of genes direct the HCV progression towards a lymphoma rather than an hepatic disease. In this contest IFN-α therapy, a standard therapy for HCV-infection and lymphoproliferative diseases, known to be able to transiently enhance the cytotoxicity of NK-cells support the role of NK cells to counterstain HCV-related and lymphoproliferative diseases.


Efficacy of short-course antibiotic prophylaxis after penetrating intestinal injury. A prospective randomized trial.
Infection is the leading cause of morbidity and mortality occurring more than 48 hours after penetrating abdominal injury. Antibiotics are routinely administered to patients with penetrating intestinal injuries and are usually given for five days or more. We randomized 116 patients with confirmed penetrating injuries of the colon and/or small bowel to receive either 12 hours or five days of antibiotics. Age, sex, weapon, severity of injury, and other risk factors were evenly distributed between groups. Twenty-one patients (18%) developed trauma-related infections, 28 (24%) any infection, and three (2.6%) died. There were no significant differences between groups in any category of outcome. For patients with penetrating intestinal or colonic injury, a 12-hour course of antibiotics is as effective as a five-day course and has the advantage of lower cost and, theoretically, fewer side effects.


How protein recognizes ladder-like polycyclic ethers. Interactions between ciguatoxin (CTX3C) fragments and its specific antibody 10C9.
Ciguatoxins are a family of marine toxins composed of transfused polycyclic ethers. It has not yet been clarified at the atomic level on the pathogenic mechanism of these toxins or the interaction between a polycyclic ether compounds and a protein. Using the crystal structures of anti-ciguatoxin antibody 10C9 Fab in ligand-free form and in complexes with ABCD-ring (CTX3C-ABCD) and ABCDE-ring (CTX3C-ABCDE) fragments of the antigen CTX3C at resolutions of 2.6, 2.4, and 2.3 angstroms, respectively, we elucidated the mechanism of the interaction between the polycyclic ethers and the antibody. 10C9 Fab has an extraordinarily large and deep binding pocket at the center of the variable region, where CTX3C-ABCD or CTX3C-ABCDE binds longitudinally in the pocket via hydrogen bonds and van der Waals interactions. Upon antigen-antibody complexation, 10C9 Fab adjusts to the antigen fragments by means of rotational motion in the variable region. In addition, the antigen fragment lacking the E-ring induces a large motion in the constant region. Consequently, the thermostability of 10C9 Fab is enhanced by 10 degrees C upon complexation with CTX3C-ABCDE but not with CTX3C-ABCD. The crystal structures presented in this study also show that 10C9 Fab recoginition of CTX3C antigens requires molecular rearrangements over the entire antibody structure. These results further expand the fundamental understanding of the mechanism by which ladder-like polycyclic ethers are recognized and may be useful for the design of novel therapeutic agents by antibodies, marine toxins, or new diagnostic reagents for the detection and targeting of members of the polycyclic ether family.


Computational Method for Evaluating the Thermoelectric Power Factor for Organic Materials Modeled by the Holstein Model: A Time-Dependent Density Matrix Renormalization Group Formalism.
Organic/polymeric materials are of emerging importance for thermoelectric conversion. The soft nature of these materials implies strong electron-phonon coupling, often leading to carrier localization. This poses great challenges for the conventional Boltzmann transport description based on relaxation time approximation and band structure calculations. In this work, combining the Kubo formula with the finite-temperature time-dependent density matrix renormalization group (FT-TD-DMRG) in the grand canonical ensemble, we developed a nearly exact algorithm to calculate the thermoelectric power factor PF = α2 σ, where α is the Seebeck coefficient and σ is the electrical conductivity, and apply the algorithm to Holstein Hamiltonian with electron-phonon coupling to model organic materials. Our algorithm can provide a unified description covering the weak coupling limit described by the bandlike Boltzmann transport to the strong coupling hopping limit.


Motility of macrogamonts of Eimeria magna (Coccidia) in cell culture.
Amoeboid movement of spheroid or ovoid immature or mature macrogamonts within cultured Madin-Darby bovine cells usually began with the formation of a pseudopodium-like protrusion at the margin of the gamont. The protrusion enlarged as the gamont cytoplasm and nucleus moved into the protruded area. During movement, macrogamonts had an elongate shape. The body of the gamont was constricted as it moved through the cell cytoplasm or from one cell to an adjacent one. After movement had ceased, the gamont resumed its ovoid or spheroidal shape.


Impact of COVID-19 on Canadian medical oncologists and cancer care: Canadian Association of Medical Oncologists survey report.
The covid-19 pandemic has presented unprecedented professional and personal challenges for the oncology community. Under the auspices of the Canadian Association of Medical Oncologists, we conducted an online national survey to better understand the impact of the pandemic on the medical oncology community in Canada. An English-language multiple-choice survey, including questions about demographics, covid-19 risk, use of personal protective equipment (ppe), personal challenges, and chemotherapy management was distributed to Canadian medical oncologists. The survey was open from 30 March to 4 April 2020, and attracted 159 responses. More than 70% of medical oncologists expressed moderate-to-extreme concern about personally contracting covid-19 and about family members or patients (or both) contracting covid-19 from them. Despite that high level of concern, considerable variability in the use of ppe in direct cancer care was reported at the time of this survey, with 33% of respondents indicating no routine ppe use at their institutions and 69% indicating uncertainty about access to adequate ppe. Of the respondents, 54% were experiencing feelings of nervousness or anxiety on most days, and 52% were having feelings of depression or hopelessness on at least some days. Concern about aging parents or family and individual wellness represented the top personal challenges identified. The management of cancer patients has been affected, with adoption of telemedicine reported by 82% of respondents, and cessation of clinical trial accrual reported by 54%. The 3 factors deemed most important for treatment decision-making were■ cancer prognosis and anticipated benefit from treatment,■ risk of treatment toxicity during scarce health care access, and■ patient risk of contracting covid-19. This report describes the results of the first national survey assessing the impact of the covid-19 on Canadian medical oncologists and how they deliver systemic anticancer therapies. We hope that these data will provide a framework to address the challenges identified.


Intra- and intersexual differences in parasite resistance and female fitness tolerance in a polymorphic insect.
To understand host-parasite interactions, it is necessary to quantify variation and covariation in defence traits. We quantified parasite resistance and fitness tolerance of a polymorphic damselfly (Ischnura elegans), an insect with three discrete female colour morphs but with monomorphic males. We quantified sex and morph differences in parasite resistance (prevalence and intensity of water mite infections) and morph-specific fitness tolerance in the females in natural populations for over a decade. There was no evidence for higher parasite susceptibility in males as a cost of sexual selection, whereas differences in defence mechanisms between female morphs are consistent with correlational selection operating on combinations of parasite resistance and tolerance. We suggest that tolerance differences between female morphs interact with frequency-dependent sexual conflict, which maintains the polymorphism locally. Host-parasite interactions can therefore shape intra- and intersexual phenotypic divergence and interfere with sexual selection and sexual conflict.


Morphometric analysis of secondary palate development in human embryos.
Rapid shelf elevation and contact of the secondary palate and fusion reportedly occur due to a growth-related equilibrium change in the structures within the oro-nasal cavity. This study aimed to quantitatively evaluate complex three-dimensional morphological changes and their effects on rapid movements, such as shelf elevation and contact, and fusion. Morphological changes during secondary palate formation were analyzed using high-resolution digitalized imaging data (phase-contrast X-ray computed tomography and magnetic resonance images) obtained from 22 human embryonic and fetal samples. The three-dimensional images of the oro-nasal structures, including the maxilla, palate, pterygoid hamulus, tongue, Meckel's cartilage, nasal cavity, pharyngeal cavity, and nasal septum, were reconstructed manually. The palatal shelves were not elevated in all the samples at Carnegie stage (CS)21 and CS22 and in three samples at CS23. In contrast, the palatal shelves were elevated but not in contact in one sample at CS23. Further, the palatal shelves were elevated and fused in the remaining four samples at CS23 and all three samples from the early fetal period. For each sample, 70 landmarks were subjected to Procrustes and principal component (PC) analysis. PC-1 accounted for 67.4% of the extracted gross changes before and after shelf elevations. Notably, the PC-1 values of the negative and positive value groups differed significantly. The PC-2 value changed during the phases in which the change in the PC-1 value was unnaturally slow and stopped at CS22 and the first half of CS23. This period, defined as the "approach period", corresponds to the time before dynamic changes occur as the palatal shelves elevate, the tongue and mandibular tip change their position and shape, and secondary palatal shelves contact and fuse. During the "approach period", measurements of PC-2 changes showed that structures on the mandible (Meckel's cartilage and tongue) and maxilla (palate and nasal cavity) did not change positions, albeit both groups of structures appeared to be compressed anterior-posteriorly. However, during and after shelf elevation, measurements of PC-1 changes showed significant changes between maxillary and mandibular structures, particularly positioning of the shelves above the tongue and protrusion of the tongue and mandible. These results suggest an active role for Meckel's cartilage growth in repositioning the tongue to facilitate shelf elevation. The present data representing three distinct phases of secondary palate closure in humans can advance the understanding of morphological growth changes occurring before and after the horizontal positioning of palatal shelves and their fusion to close the secondary palate in humans successfully.


[Intensive combined chemo- and radiotherapy in small cell bronchial carcinoma].
117 patients with small cell bronchogenic carcinoma stage III-IV were treated, 82 patients with a combined chemotherapy and radiotherapy and 35 patients with combined chemotherapy only. All schedules investigated were successful. Long term remissions were obtained only in patients with a relatively located tumor, who achieved complete remission in consequence of treatment. Therapy results will be improved in future by perfecting the existing methods of treatment.


Revisiting gap locations in amino acid sequence alignments and a proposal for a method to improve them by introducing solvent accessibility.
In comparative modeling, the quality of amino acid sequence alignment still constitutes a major bottleneck in the generation of high quality models of protein three-dimensional (3D) structures. Substantial efforts have been made to improve alignment quality by revising the substitution matrix, introducing multiple sequences, replacing dynamic programming with hidden Markov models, and incorporating 3D structure information. Improvements in the gap penalty have not been a major focus, however, following the development of the affine gap penalty and of the secondary structure dependent gap penalty. We revisited the correlation between protein 3D structure and gap location in a large protein 3D structure data set, and found that the frequency of gap locations approximated to an exponential function of the solvent accessibility of the inserted residues. The nonlinearity of the gap frequency as a function of accessibility corresponded well to the relationship between residue mutation pattern and residue accessibility. By introducing this relationship into the gap penalty calculation for pairwise alignment between template and target amino acid sequences, we were able to obtain a sequence alignment much closer to the structural alignment. The quality of the alignments was substantially improved on a pair of sequences with identity in the "twilight zone" between 20 and 40%. The relocation of gaps by our new method made a significant improvement in comparative modeling, exemplified here by the Bacillus subtilis yitF protein. The method was implemented in a computer program, ALAdeGAP (ALignment with Accessibility dependent GAp Penalty), which is available at http://cib.cf.ocha.ac.jp/target_protein/.


The Normativity of Law: Has the Dispositional Model Solved our Problem?
In Legal Directives and Practical Reasons, Noam Gur has presented a novel account, called the dispositional model, to explain how law bears on our normative practical reasons. Gur holds that his model is superior to the current models, namely the standard weighing model and Joseph Raz's exclusionary model. Although his work provides useful insights into the practical impact of law, I argue that: (i) his challenge against the exclusionary model is valid only insofar as one accepts Raz's normal justification thesis and dependence thesis; (ii) his argument against the weighing model misses its target, because it attacks the model as a decision-making method, not as an account of practical reason; and (iii) his dispositional model solely constitutes a decision-making strategy and does not offer a third alternative answer to the question of how law affects our normative practical reasons. Hence, the dispositional model is not a competitor to the weighing and the exclusionary model, and the problem of accounting for the normativity of law remains.


Clinical comparison of the Proview eye pressure monitor with the Goldmann applanation tonometer and the Tonopen.
To compare intraocular pressure (IOP) values obtained by patients using the new Proview eye pressure monitor (Bausch & Lomb, Rochester, NY) with those measured with the Goldmann tonometer and the TonoPen (Mentor, Norwell, Mass). Eighty-six patients (a total of 171 eyes) with a diagnosis of glaucoma or glaucoma suspect successfully completed the study. The IOP was measured by 3 methods in the following order: Goldmann tonometer, TonoPen, and Proview eye pressure monitor. The central corneal thickness was measured by an ultrasonic pachymeter. Separately for each eye, the differences in mean IOP values between measurement methods were assessed with paired t tests and also in multivariate models that tested the dependence of IOP difference on central corneal thickness. There was a significant difference (P<.001) in the mean IOPs measured by the 3 different methods (Goldmann vs Proview, Goldmann vs TonoPen, and TonoPen vs Proview) for both eyes, and the difference was independent of the central corneal thickness. The differences between IOP measured by Goldmann and Proview were similar in all categories of patient-reported ease of using the Proview. The IOPs obtained with the Proview eye pressure monitor are significantly lower than those measured with Goldmann tonometer and the TonoPen, and variations of the central corneal thickness do not contribute to the difference. Intraclass correlations of IOP values obtained with the Goldmann and the Proview or TonoPen and Proview are not strong. On the other hand, as expected, measurements with Goldmann and TonoPen agreed fairly well.


The role of thymidylate synthase induction in modulating p53-regulated gene expression in response to 5-fluorouracil and antifolates.
Thymidylate synthase (TS) is a critical target for chemotherapeutic agents such as 5-fluorouracil (5-FU) and antifolates such as tomudex (TDX),multitargeted antifolate, and ZD9331. Using the MCF-7 breast cancer line, we have developed p53 wild-type (M7TS90) and null (M7TS90-E6) isogenic lines with inducible TS expression (approximately 6-fold induction compared with control after 48 h). In the M7TS90 line, inducible TS expression resulted in a moderate approximately 3-fold increase in 5-FU IC-50(72 h) dose and a dramatic >20-fold increase in the IC-50(72 h) doses of TDX, multitargeted antifolate, and ZD9331. S-phase cell cycle arrest and apoptosis induced by the antifolates were abrogated by TS induction. In contrast, cell cycle arrest and apoptosis induced by 5-FU was unaffected by TS expression levels. Inactivation of p53 significantly increased resistance to 5-FU and the antifolates with IC-50(72 h) doses for 5-FU and TDX of >100 and >10 microM, respectively, in the M7TS90-E6 cell line. Furthermore, p53 inactivation completely abrogated the cell cycle arrest and apoptosis induced by 5-FU. The antifolates induced S-phase arrest in the p53 null cell line; however, the induction of apoptosis by these agents was significantly reduced compared with p53 wild-type cells. Both inducible TS expression and the addition of exogenous thymidine (10 microM) blocked p53 and p21 induction by the antifolates but not by 5-FU in the M7TS90 cell line. Similarly, inducible TS expression and exogenous thymidine abrogated antifolate but not 5-FU-mediated up-regulation of Fas/CD95 in M7TS90 cells. Our results indicate that in M7TS90 cells, inducible TS expression modulates p53 and p53 target gene expression in response to TS-targeted antifolate therapies but not to 5-FU.


A comparative study of convicilin storage protein gene sequences in species of the tribe Vicieae.
Convicilins, a set of seed storage proteins, differ from vicilins, a related group of seed storage proteins, mainly because of the presence of the N-terminal extension, an additional sequence of amino acids in the sequence corresponding to the first exon. Convicilins have been described only in species of the legume tribe Vicieae. One or two genes for convicilins have been identified in most species of this tribe. The genus Pisum is the main exception, since two genes have been identified in most of its species. Thirty-four new convicilin gene sequences from 29 different species (Lathyrus, Lens, Pisum, and Vicia spp.) have been analyzed here. Convicilin gene sequences are generally organized in 6 exons, but in some instances one of the internal introns (2nd or 4th) is lost. In these 29 species, the N-terminal extension is formed by a stretch of 99 to 196 amino acids particularly rich in polar and charged amino acids (on average, it contains 29.43% glutamic acid and 15.38% arginine residues). This N-terminal extension has the characteristics of an intrinsically unstructured region (IUR), one of the categories of protein "degenerate sequences". A comparative analysis indicates that the N-terminal extension sequence has evolved faster than the surrounding sequence, which is common to all vicilins, and it evolved mainly through a series of duplications of short internal sequences and triplet expansions, the predominant one being GAA. This agrees with the evolution of IURs, which is faster than the evolution of surrounding sequences and is mainly due to replication slippage and unequal crossover recombination. Alternative maximum-likelihood trees of phylogenetic relationships among the 29 Vicieae species based on the convicilin exon sequences are presented and discussed.


Does a patient with hereditary spherocytosis qualify for preoperative autologous blood donation?
Hereditary spherocytosis (HS) is characterized by osmotically fragile spherocytic red cells (RBCs), mild to moderate hemolysis, and splenomegaly. Little is known about the safety of blood bank storage of HS RBCs. A 50-year-old man with mild autosomally dominant HS asked to make an autologous RBC deposit before his scheduled surgery. His RBCs were serially tested for osmotic fragility during 4 weeks of blood bank storage in CPD with adenine-saline. Aliquots of his fresh and 4-week-old RBCs were also labeled with 51Cr for measurement of in vivo survival. The osmotic fragility of the patient's fresh RBCs was normal, with 50-percent hemolysis at approximately 0.43-percent NaCl and a sigmoid lysis curve. Incubated osmotic fragility (24 hours at 37 degrees C) was abnormal, with 50-percent hemolysis at 0.64-percent NaCl. Median survival of his fresh 51Cr-labeled RBCs was moderately shortened at 17.5 days. Osmotic fragility increased during storage, with 50-percent lysis occurring at 0.58-percent and 0.62-percent NaCl after 2 and 4 weeks, respectively. Volunteer normal donor RBCs exhibited 50-percent lysis in 0.48-percent NaCl after 4 weeks. Median survival of the patient's 4-week-old 51Cr-labeled RBCs was severely shortened, at 3.5 days. At surgery, intraoperatively salvaged RBCs demonstrated osmotic fragility identical to that of the patient's RBCs freshly obtained by venipuncture. HS RBCs may lose membrane under optimum storage conditions, becoming unsuitable for transfusion at surgery. Intraoperative autologous transfusion may be preferable to preoperative deposit for avoidance of allogeneic RBC transfusion in patients with HS.


Stability of and attachment to lettuce by a culturable porcine sapovirus surrogate for human caliciviruses.
Human noroviruses (HuNoVs) are the leading cause of food-borne illness, accounting for 58% of U.S. cases. Because HuNoVs are unculturable, surrogates are needed to investigate transmission routes and evaluate disinfection methods. However, the current surrogates, feline calicivirus (FCV) and murine NoV (MNV), are less tolerant than HuNoVs to acid and chlorine, respectively. Porcine sapovirus (SaV) is the only culturable enteropathogenic calicivirus. In this study, the resistance of SaV to physicochemical treatments was compared to that of HuNoVs (by reverse transcription-PCR), FCV, and MNV (by infectivity assays). Sapovirus and HuNoV (viral RNA) showed similar resistances to heat (56°C) and to different concentrations of chlorine. However, SaV was more resistant than HuNoVs to ethanol treatment (60% and 70%). Like HuNoVs, SaV was stable at pH 3.0 to 8.0, with a <1.0 log(10) 50% tissue culture infective dose (TCID(50)) reduction at pH 3.0 compared to the value for pH 4.0 to 8.0. SaV and MNV showed similar resistances, and both were more resistant than FCV to heat inactivation (56°C). FCV was more resistant than MNV and SaV to ethanol, and all three viruses showed similar resistances to treatment with low concentrations of chlorine for 1 min. Those results indicate that SaV is a promising surrogate for HuNoVs. Next, we used SaV as a surrogate to examine virus attachment to lettuce at different pHs. Sapovirus attached to lettuce leaves significantly at its capsid isoelectric point (pH 5.0), and the attached viral particles remained infectious on lettuce after 1 week of storage at 4°C. The culturable SaV is a good surrogate for studying HuNoV contamination and transmission in leafy greens and potential disinfectants.