Genotypic characteristics of Staphylococcus aureus isolates from a multinational trial of complicated skin and skin structure infections.
The impact of bacterial genetic characteristics on the outcome of patients with Staphylococcus aureus infections is uncertain. This investigation evaluated potential associations between bacterial genotype and clinical outcome using isolates collected as part of an international phase 2 clinical trial (FAST II) evaluating telavancin for the treatment of complicated skin and skin structure infections (cSSSI). Ninety S. aureus isolates from microbiologically evaluable patients with cSSSI enrolled in the FAST II trial from 11 sites in the United States (56 isolates, or 62%) and 7 sites in South Africa (34 isolates, or 38%) were examined for staphylococcal cassette chromosome mec, agr, and the presence of 31 virulence genes and subjected to pulsed-field gel electrophoresis (PFGE). South African methicillin-susceptible S. aureus (MSSA) isolates were more likely to carry certain virulence genes, including sdrD (P = 0.01), sea (P < 0.01), and pvl (P = 0.01). All 44 (49%) methicillin-resistant S. aureus (MRSA) isolates were from the United States; 37 (84%) were strain USA 300 by PFGE. In the United States, MRSA isolates were more likely than MSSA isolates to carry genes for sdrC (P = 0.03), map/eap (P = 0.05), fnbB (P = 0.11), tst (P = 0.02), sea (P = 0.04), sed (P = 0.04), seg (P = 0.11), sej (P = 0.11), agr (P = 0.09), V8 (P = 0.06), sdrD, sdrE, eta, etb, and see (P < 0.01 for all). MRSA isolates were more often clonal than MSSA isolates by PFGE. Isolates from patients who were cured were significantly more likely to contain the pvl gene than isolates from patients that failed or had indeterminate outcomes (79/84 [94%] versus 3/6 [50%]; P = 0.01). S. aureus strains from different geographic regions have different distributions of virulence genes.


Development and evaluation of a drop-and-stick method to assess landing skills in various levels of competitive surfers.
The purpose of this study was to develop and evaluate a drop-and-stick (DS) test method and to assess dynamic postural control in senior elite (SE), junior elite (JE), and junior development (JD) surfers. Nine SE, 22 JE, and 17 JD competitive surfers participated in a single testing session. The athletes completed 5 drop-and-stick trials barefoot from a predetermined box height (0.5 m). The lowest and highest time-to-stabilization (TTS) trials were discarded, and the average of the remaining trials was used for analysis. The SE group demonstrated excellent single-measures repeatability (ICC = .90) for TTS, whereas the JE and JD demonstrated good single-measures repeatability (ICC .82 and .88, respectively). In regard to relative peak landing force (rPLF), SE demonstrated poor single-measures reliability compared with JE and JD groups. Furthermore, TTS for the SE (0.69 ± 0.13 s) group was significantly (P = .04) lower than the JD (0.85 ± 0.25 s). There were no significant (P = .41) differences in the TTS between SE (0.69 ± 0.13 s) and JE (0.75 ± 0.16 s) groups or between the JE and JD groups (P = .09). rPLF for the SE (2.7 ± 0.4 body mass; BM) group was significantly lower than the JE (3.8 ± 1.3 BM) and JD (4.0 ± 1.1 BM), with no significant (P = .63) difference between the JE and JD groups. A possible benchmark approach for practitioners would be to use TTS and rPLF as a qualitative measure of dynamic postural control using a reference scale to discriminate among groups.


Proton-coupled electron transfer of the phenoxyl/phenol couple: effect of Hartree-Fock exchange on transition structures.
Proton-coupled electron transfer (PCET) and hydrogen atom transfer (HAT) reactions of the phenoxyl/phenol couple are studied theoretically by using wave function theory (WFT) as well as DFT methods. At the complete active space self-consistent field (CASSCF) level, geometry optimization is found to give two transition states (TSs); one is the PCET type with two benzene rings being nearly coplanar, and the other is the HAT type with two benzene rings taking a stacking structure. Geometry optimization at the (semilocal) DFT level, on the other hand, is found to give only one transition state (i.e., the PCET-type one) and fail to obtain the stacking TS structure. By comparing various levels of theories (including long-range corrected DFT functionals), we demonstrate that the Hartree-Fock exchange at long range plays a critical role in obtaining the sufficient stacking stabilization of the present open-shell system, and that the sole addition of empirical dispersion correction to semilocal DFT functionals may not be adequate for describing such a stacking interaction. Next, we investigate the solvent effect on the PCET and HAT TS thus obtained using the reference interaction site model self-consistent field (RISM-SCF) method. The results suggest that the free energy barrier increases with increasing polarity of the solvent, and that the solvent effects are stronger for the PCET TS than the stacking HAT TS pathway. The reason for this is discussed based on the dipole moment of different TS structures in solution.


The Longitudinal Parallel Process Analysis of Biomarkers of Oxidative Stress, Symptom Clusters, and Cognitive Function in Children With Leukemia.
Background: During treatment for acute lymphoblastic leukemia (ALL), children report co-occurring symptoms of fatigue, sleep disturbance, pain, nausea, and depression as a symptom cluster. Central nervous system-directed ALL therapies also put children at risk for cognitive impairments. Cancer therapies can cause an increase in oxidative stress, which may contribute to treatment-related symptoms. This study examined the longitudinal relationships between biomarkers of oxidative stress in the cerebrospinal fluid, the Childhood Cancer Symptom Cluster-Leukemia (CCSC-L), and cognition, in children over the first year of ALL treatment. Methods: Glutathione (GSH) biomarkers of oxidative stress were measured in cerebrospinal fluid collected during treatment lumbar punctures. GSH biomarkers, symptoms, and cognitive function of 132 children aged 3 to 18 years were evaluated at four time points during the first year of leukemia treatment. Participants, 7 years and older, completed self-report measures, and parents reported for younger children. Cognitive function measurements for all participants were completed by parents. A longitudinal parallel-process model was used to explore the influence of the initial measurement and the subsequent change over four time points of the GSH biomarkers on the CCSC-L and cognition. Results: GSH biomarkers increased over the four time points indicating decreasing oxidative stress. When GSH biomarkers were higher (less oxidative stress) at the initial measurement, the CCSC-L severity was lower, cognition was better, and cognition improved over the four measurements. Screening children for high levels of oxidative stress would be a foundation for future intervention studies to address symptom distress and cognitive impairments.


Reduced ipsilesional cortical volumes in fetal periventricular venous infarction.
Perinatal stroke causes most term-born hemiplegic cerebral palsy. Many suffer additional sequelae. Periventricular venous infarction (PVI) is a common fetal stroke in which isolated subcortical injury may cause only motor deficits. However, cognitive, language, and behavioral deficits also occur. We hypothesized that ipsilesional cortical gray matter volumes are reduced in PVI. Children (12 months to 18 years) with MRI-confirmed PVI were identified through the Alberta Perinatal Stroke Project. We developed an MRI method to quantify sectional gray (GM) and white matter (WM) volumes from lesioned and unlesioned (control) hemispheres (OsiriX software). Differences in cortical GM and WM volumes were compared between hemispheres in preselected regions "above" the lesion (middle) and anterior and posterior to this. Outcomes dichotomized for "cortical dysfunction" (cognitive, behavioral, language) and motor deficit severity (Pediatric Stroke Outcome Measure) were compared with GM volumes. Twenty-two children (81% boys; median age, 8 years) were included. Methods demonstrated high intrarater and inter-rater reliabilities (ρ=0.988, ρ=0.943) and minimal observer bias. Ipsilesional GM volume was significantly reduced in the middle (P=0.007) and posterior (P=0.03) regions. Middle ipsilesional WM volumes were reduced (P<0.001). The degree of GM reduction was not associated with cortical dysfunction or severity of motor deficit. Ipsilesional GM volume is diminished in PVI. Speculative mechanisms include retrograde neuronal degeneration and disrupted migration. Neuropsychological testing of larger samples is required to determine clinical significance.


Effect of playing violent video games cooperatively or competitively on subsequent cooperative behavior.
Research on video games has yielded consistent findings that violent video games increase aggression and decrease prosocial behavior. However, these studies typically examined single-player games. Of interest is the effect of cooperative play in a violent video game on subsequent cooperative or competitive behavior. Participants played Halo II (a first-person shooter game) cooperatively or competitively and then completed a modified prisoner's dilemma task to assess competitive and cooperative behavior. Compared with the competitive play conditions, players in the cooperative condition engaged in more tit-for-tat behaviors-a pattern of behavior that typically precedes cooperative behavior. The social context of game play influenced subsequent behavior more than the content of the game that was played.


Cancer-Associated Fibroblasts Induce a Collagen Cross-link Switch in Tumor Stroma.
Intratumoral collagen cross-links heighten stromal stiffness and stimulate tumor cell invasion, but it is unclear how collagen cross-linking is regulated in epithelial tumors. To address this question, we used Kras(LA1) mice, which develop lung adenocarcinomas from somatic activation of a Kras(G12D) allele. The lung tumors in Kras(LA1) mice were highly fibrotic and contained cancer-associated fibroblasts (CAF) that produced collagen and generated stiffness in collagen gels. In xenograft tumors generated by injection of wild-type mice with lung adenocarcinoma cells alone or in combination with CAFs, the total concentration of collagen cross-links was the same in tumors generated with or without CAFs, but coinjected tumors had higher hydroxylysine aldehyde-derived collagen cross-links (HLCC) and lower lysine-aldehyde-derived collagen cross-links (LCCs). Therefore, we postulated that an LCC-to-HLCC switch induced by CAFs promotes the migratory and invasive properties of lung adenocarcinoma cells. To test this hypothesis, we created coculture models in which CAFs are positioned interstitially or peripherally in tumor cell aggregates, mimicking distinct spatial orientations of CAFs in human lung cancer. In both contexts, CAFs enhanced the invasive properties of tumor cells in three-dimensional (3D) collagen gels. Tumor cell aggregates that attached to CAF networks on a Matrigel surface dissociated and migrated on the networks. Lysyl hydroxylase 2 (PLOD2/LH2), which drives HLCC formation, was expressed in CAFs, and LH2 depletion abrogated the ability of CAFs to promote tumor cell invasion and migration. CAFs induce a collagen cross-link switch in tumor stroma to influence the invasive properties of tumor cells.


Effect of Low-Frequency rTMS and NMES on Subacute Unilateral Hemispheric Stroke With Dysphagia.
To investigate the effect of low-frequency repetitive transcranial magnetic stimulation (rTMS) and neuromuscular electrical stimulation (NMES) on post-stroke dysphagia. Subacute (<3 months), unilateral hemispheric stroke patients with dysphagia were randomly assigned to the conventional dysphagia therapy (CDT), rTMS, or NMES groups. In rTMS group, rTMS was performed at 100% resting motor threshold with 1 Hz frequency for 20 minutes per session (5 days per week for 2 weeks). In NMES group, electrical stimulation was applied to the anterior neck for 30 minutes per session (5 days per week for 2 weeks). All three groups were given conventional dysphagia therapy for 4 weeks. We evaluated the functional dysphagia scale (FDS), pharyngeal transit time (PTT), the penetration-aspiration scale (PAS), and the American Speech-Language Hearing Association National Outcomes Measurement System (ASHA NOMS) swallowing scale at baseline, after 2 weeks, and after 4 weeks. Forty-seven patients completed the study; 15 in the CDT group, 14 in the rTMS group, and 18 in the NMES group. Mean changes in FDS and PAS for liquid during first 2 weeks in the rTMS and NMES groups were significantly higher than those in the CDT group, but no significant differences were found between the rTMS and NMES group. No significant difference in mean changes of FDS and PAS for semi-solid, PTT, and ASHA NOMS was observed among the three groups. These results indicated that both low-frequency rTMS and NMES could induce early recovery from dysphagia; therefore, they both could be useful therapeutic options for dysphagic stroke patients.


On-site HIV testing in residential drug treatment units: results of a nationwide survey.
Residential drug treatment units are uniquely situated to provide HIV testing and counseling to their patients. This article examines the extent to which residential drug treatment units in the United States provide HIV testing on-site, and identifies organizational and institutional characteristics that differentiate units in which on-site HIV testing is available from those in which it is not. The analyses use data collected in telephone interviews with unit managers from a random nationwide sample (N = 138) of residential drug treatment units in 2001. About half (48.6%) of the residential drug treatment units made HIV testing available to their patients on-site. Residential units were significantly more likely to make on-site testing available if they were larger (i.e., had a greater number of patients treated each month or had a greater number of staff that provided direct patient services) and if they were publicly rather than privately owned. Provision of on-site HIV testing was significantly correlated with having a medical orientation, i.e., with being operated by a hospital, with the unit viewing itself as patients' primary medical provider, or with providing medical care to the patients either on-site or at another part of the same treatment agency. In view of the critical importance of HIV testing for individuals who use illicit drugs and the existence of a simplified testing protocol involving saliva samples (eliminating the need for phlebotomy), units that do not have a medical orientation should be encouraged to make HIV testing available on-site.


Small Molecule Targeting of Oxysterol-Binding Protein (OSBP)-Related Protein 4 and OSBP Inhibits Ovarian Cancer Cell Proliferation in Monolayer and Spheroid Cell Models.
The development of precision drugs for the selective treatment of ovarian cancer will require targeting proliferative factors selectively expressed in ovarian tumors or targeting unique physiological microenvironments specific for ovarian tumors. Here, we report that oxysterol-binding protein (OSBP)-related protein 4 (ORP4) is a potential druggable precision target in ovarian cancer cells. ORP4 has limited expression in normal tissues and was recently recognized to be a cancer-specific driver of cellular proliferation, including in patient-isolated leukemias. We demonstrate that ORP4 is strongly expressed in a panel of ovarian cancer cell lines. The antiproliferative natural product compound OSW-1 targets ORP4 and OSBP. Our results demonstrate that the OSW-1 compound has high antiproliferative potency in both monolayer and three-dimensional ovarian cancer spheroid models, especially compared to the standard-of-care agents cisplatin and paclitaxel. OSW-1 compound treatment induces a loss of ORP4 expression after 48 h, which is coincident with the cytotoxic effects of OSW-1. The absence of extracellular lipids markedly potentiated the cytotoxicity of OSW-1, which was reversed by addition of extracellular free cholesterol. OSBP, but not ORP4, is reported to transport cholesterol and other lipids between organelles. Our results indicate that the targeting of ORP4 is responsible for the antiproliferative activity of the OSW-1 compound, but that in the absence of exogenously supplied cholesterol, which might be similar to the in vivo ovarian cancer microenvironment, possible OSW-1 targeting of OSBP further potentiates the anticancer activity of the compound. Overall, ORP4 and potentially OSBP are revealed as potential druggable targets for the development of novel treatments for ovarian cancer.


Electrostatic interactions during Kv1.2 N-type inactivation: random-walk simulation.
N-type inactivation of the Kv1.2 voltage-gated potassium channel is a process in which the N-terminal of the protein (its first 20 amino acids) binds to the open-channel surface, extends and occludes its pore. This process has been experimentally studied in both intact and ShBDelta6-46 channels in which the inactivating peptides are supplied in the bath solution. In this work we provide a qualitative description of N-type inactivation by simulating the random walk of charged inactivating peptides in the electrostatic field that originates from the charges present in the channel and in the cellular membrane. Our results give a deeper insight into the previously reported influence of electrostatics on the rate of N-type inactivation of ShBDelta6-46. We also show how the enchaining of the peptides, i.e., considering the intact form of the channel, influences the N-type inactivation with different charges of those peptides.


Supervision trajectories of male juvenile offenders: growth mixture modeling on SAVRY risk assessments.
Structured risk/need assessment tools are increasingly used to orientate risk reduction strategies with juvenile offenders. The assumption is that the risk/need items on these tools are sufficiently sensitive to measure changes in the individual, family and/or contextual characteristics of juvenile offenders. However, there is very little research demonstrating the capacity of these tools to measure changes in juvenile offenders. Congruent with the developmental and life-course criminology theories (DLC) the objective of this study is to explore the existence of heterogeneous trajectories of juvenile offenders across the juvenile justice system as measured through five empirical risk/need areas based on the Structured Assessment of Violence Risk in Youth (SAVRY), one of the most widely applied risk assessment tools for juveniles. This longitudinal study included 5205 male juvenile offenders who transitioned through the Catalan juvenile justice system between 2006 and 2014. During intervention they received at least two, and a maximum of seven, consecutive SAVRY risk/need assessments over an 18-month period. The heterogeneity of latent class trajectories was explored through growth mixture modeling (GMM). The trajectory class membership was linked to covariates through multinomial logistic regression analyses. Through GMM three to four heterogeneous trajectories, with high quality of separation, were identified in each of the risk/need areas. The trajectories with low risk/needs (45-77% of the sample) remained low and presented a very limited increase in risk/needs during the 18-month period. The high risk/need trajectories (20-37% of the sample) showed a limited decrease or no change. Between 5 and 13% of the sample had large reductions in their risk/needs levels, and approximately 5% showed a large increase in risk/needs. In line with the DLC theories this study shows that trajectories on criminogenic risk/needs can be heterogeneous and indicate distinct rates of change over time. The results of this study also may suggest a limited sensibility to measure change over time of SAVRY's risk and protective items. Suggestions to improve the sensitivity of measuring change over time, such as shorter time frames or future-oriented time frames for the scoring of the items, are offered.


Systematic Review and Meta-Analyses: Motor Skill Interventions to Improve Fine Motor Development in Children Aged Birth to 6 Years.
The objective of this study was to systematically review the evidence of motor skill intervention programs on typically developing children's fine motor development aged birth to 6 years. Six electronic databases were searched with no date restrictions. Inclusion criteria were any school-, community-, or home-based intervention targeting the development of fine motor skills of children aged birth to 6 years; randomized controlled trials using quasiexperimental, experimental, or single group prepost designs with a minimum sample size of 15 participants per group; and statistical analyses of fine motor skill development at both preintervention and postintervention or addressing the intervention effects on fine motor skill development. Data were extracted on design, participants, intervention components, methodological quality, and efficacy. Twenty-five of the total 31 studies reported positive intervention effects on fine motor skills. The meta-analyses included 19 studies and revealed moderate effect sizes of motor skill programs on fine motor, visual motor, and manual dexterity outcomes. There were substantial differences between intervention settings, facilitators, length, and content with most studies implemented in school settings and facilitated by teachers. Fine motor skill development in the early years is an extensive upcoming field of interest for many international researchers. This review study presents evidence on the positive effects of intervention programs that aim to enhance fine motor skills for young children. The findings are promising but need to be interpreted with caution because of the high risk of bias in many of the studies.


UK internal medicine training in the time of COVID-19.
COVID-19 has had a significant impact on internal medicine training in the UK. Many unprecedented changes have been made to prioritise the care of affected patients. The medical workforce was re-shaped, training programmes were disrupted, Membership of the Royal College of Physicians (MRCP) examinations were cancelled and their format changed on re-commencement, teaching programmes were suspended and delivery methods amended, out of programme (OOP) opportunities deferred, non-COVID related research halted, trainee progression impacted and trainee mental health and wellbeing suffered. Despite this, the pandemic has undoubtedly created a plethora of unique learning opportunities for trainees that could make them better doctors and healthcare leaders in the future.


Edar and Troy signalling pathways act redundantly to regulate initiation of hair follicle development.
The development of ectodermal organs requires signalling by ectodysplasin (Eda), a tumor necrosis factor (TNF) family member, its receptor Edar and downstream activation of the nuclear factor kappaB (NF-kappaB) transcription factor. In humans, mutations in the Eda pathway components cause hypohidrotic ectodermal dysplasia, a syndrome characterized by missing teeth, sparse hair and defects in sweat glands. It has been postulated that Eda acts redundantly with another TNF pathway to regulate ectodermal organogenesis. A potential candidate is Troy (or TNFRSF19 or Taj), a TNF receptor which is homologous with Edar in its ligand-binding domain, and is expressed in an overlapping pattern. We have characterized Troy null mice and crossed them with Eda-deficient mice. Single Troy mutants had no defects in ectodermal organs. Analysis of the double mutants revealed an essential role for Troy in hair follicle development. In mice, hair follicles develop in three different waves. Only primary hair follicles are missing in Eda single mutants, whereas the compound mutants lacked also the follicles of the second wave, as well as all hair follicles in the middle of crown leading to focal alopecia. Assessment of NF-kappaB activity with a transgenic reporter construct indicated that Eda is the main activator of NF-kappaB signalling in developing skin appendages and surprisingly that the functional overlap of Troy and Eda signalling pathways is mediated by NF-kappaB independent pathways.


Interactions of innate and adaptive immunity in brain development and function.
It has been known for decades that the immune system has a tremendous impact on behavior. Most work has described the negative role of immune cells on the central nervous system. However, we and others have demonstrated over the last decade that a well-regulated immune system is needed for proper brain function. Here we discuss several neuro-immune interactions, using examples from brain homeostasis and disease states. We will highlight our understanding of the consequences of malfunctioning immunity on neurodevelopment and will discuss the roles of the innate and adaptive immune system in neurodevelopment and how T cells maintain a proper innate immune balance in the brain surroundings and within its parenchyma. Also, we describe how immune imbalance impairs higher order brain functioning, possibly leading to behavioral and cognitive impairment. Lastly, we propose our hypothesis that some behavioral deficits in neurodevelopmental disorders, such as in autism spectrum disorder, are the consequence of malfunctioning immunity. This article is part of a Special Issue entitled SI: Neuroimmunology in Health And Disease.


Positional interchanges influence the physical and technical match performance variables of elite soccer players.
Positional variation in match performance is well established in elite soccer but no information exists on players switching positions. This study investigated the influence of elite players interchanging from one position to another on physical and technical match performance. Data were collected from multiple English Premier League (EPL) seasons using a computerised tracking system. After adhering to stringent inclusion criteria, players were examined across several interchanges: central-defender to fullback (CD-FB, n = 11, 312 observations), central-midfielder to wide-midfielder (CM-WM, n = 7, 171 observations), wide-midfielder to central-midfielder (WM-CM, n = 7, 197 observations) and attacker to wide-midfielder (AT-WM, n = 4, 81 observations). Players interchanging from CD-FB covered markedly more high-intensity running and sprinting distance (effect size [ES]: -1.56 and -1.26), lost more possessions but made more final third entries (ES: -1.23 and -1.55). Interchanging from CM-WM and WM-CM resulted in trivial to moderate differences in both physical (ES: -0.14-0.59 and -0.21-0.39) and technical performances (ES: -0.48-0.64 and -0.36-0.54). Players interchanging from AT-WM demonstrated a moderate difference in high-intensity running without possession (ES: -0.98) and moderate-to-large differences in the number of clearances, tackles and possessions won (ES: -0.77, -1.16 and -1.41). The data demonstrate that the physical and technical demands vary greatly from one interchange to another but utility players seem able to adapt to these positional switches.


Requirement for coronin 1 in T lymphocyte trafficking and cellular homeostasis.
The evolutionarily conserved actin-related protein (Arp2/3) complex is a key component of actin filament networks that is dynamically regulated by nucleation-promoting and inhibitory factors. Although much is known about actin assembly, the physiologic functions of inhibitory proteins are unclear. We generated coronin 1-/- mice and found that coronin 1 exerted an inhibitory effect on cellular steady-state F-actin formation via an Arp2/3-dependent mechanism. Whereas coronin 1 was required for chemokine-mediated migration, it was dispensable for T cell antigen receptor functions in T cells. Moreover, actin dynamics, through a mitochondrial pathway, was linked to lymphocyte homeostasis.


Human pluripotent reprogramming with CRISPR activators.
CRISPR-Cas9-based gene activation (CRISPRa) is an attractive tool for cellular reprogramming applications due to its high multiplexing capacity and direct targeting of endogenous loci. Here we present the reprogramming of primary human skin fibroblasts into induced pluripotent stem cells (iPSCs) using CRISPRa, targeting endogenous OCT4, SOX2, KLF4, MYC, and LIN28A promoters. The low basal reprogramming efficiency can be improved by an order of magnitude by additionally targeting a conserved Alu-motif enriched near genes involved in embryo genome activation (EEA-motif). This effect is mediated in part by more efficient activation of NANOG and REX1. These data demonstrate that human somatic cells can be reprogrammed into iPSCs using only CRISPRa. Furthermore, the results unravel the involvement of EEA-motif-associated mechanisms in cellular reprogramming.


Molecular diversity of L-type calcium channels. Evidence for alternative splicing of the transcripts of three non-allelic genes.
The diversity of L-type calcium channels was probed using the polymerase chain reaction and primers based on regions conserved in the L-type skeletal muscle (CaCh 1) and cardiac calcium channels (CaCh 2). Related sequences were amplified from human heart, hamster heart, rabbit heart, mouse ovary, mouse BC3H1 cells, and hamster insulin-secreting (HIT) cells. Sequencing of various clones revealed the presence of alternate splicing in gene products coding for CaCh 1, CaCh 2, and a related calcium channel. This related gene product, which we refer to as neuroendocrine or CaCh 3, is expressed in brain and endocrine cells. The diverse products can be explained by the use of alternate exons of equal size, which account for changes in amino acid composition, in combination with an alternate splice acceptor site or an exon skipping event, which produces channels of variable length. Four variants were defined for the gene 3 product, subtypes 3a, 3b, 3c, and 3d that differed in both the sequence of the third membrane spanning segment of the fourth repeat unit (IVS3) and in the size of the linker between this and the fourth membrane spanning segment (IVS4). Three CaCh 2 variants were cloned, subtypes 2a, 2c, and 2d, that are homologous to the a, c, and d variants of CaCh 3. For the skeletal muscle calcium channel only two variants were isolated. They are homologous to those of the a and c subtypes of CaCh 2 or 3, in that they differ only in the size of the IVS3 to IVS4 linker. These results demonstrate that calcium channel diversity is created by both the expression of distinct genes and the alternate splicing of these genes.


A Review of Spectroscopic and Biophysical-Chemical Studies of the Complex of Cyclobutane Pyrimidine Dimer Photolyase and Cryptochrome DASH with Substrate DNA.
Cyclobutane pyrimidine dimer (CPD) photolyase (PL) is a structure-specific DNA repair enzyme that uses blue light to repair CPD on DNA. Cryptochrome (CRY) DASH enzymes use blue light for the repair of CPD lesions on single-stranded (ss) DNA, although some may also repair these lesions on double-stranded (ds) DNA. In addition, CRY DASH may be involved in blue light signaling, similar to cryptochromes. The focus of this review is on spectroscopic and biophysical-chemical experiments of the enzyme-substrate complex that have contributed to a more detailed understanding of all the aspects of the CPD repair mechanism of CPD photolyase and CRY DASH. This will be performed in the backdrop of the available X-ray crystal structures of these enzymes bound to a CPD-like lesion. These structures helped to confirm conclusions that were drawn earlier from spectroscopic and biophysical-chemical experiments, and they have a critical function as a framework to design new experiments and to interpret new experimental data. This review will show the important synergy between X-ray crystallography and spectroscopic/biophysical-chemical investigations that is essential to obtain a sufficiently detailed picture of the overall mechanism of CPD photolyases and CRY DASH proteins.


Preeclampsia is associated with compromised maternal synthesis of long-chain polyunsaturated fatty acids, leading to offspring deficiency.
Obesity and excessive lipolysis are implicated in preeclampsia (PE). Intrauterine growth restriction is associated with low maternal body mass index and decreased lipolysis. Our aim was to assess how maternal and offspring fatty acid metabolism is altered in mothers in the third trimester of pregnancy with PE (n=62) or intrauterine growth restriction (n=23) compared with healthy pregnancies (n=164). Markers of lipid metabolism and erythrocyte fatty acid concentrations were measured. Maternal adipose tissue fatty acid composition and mRNA expression of adipose tissue fatty acid-metabolizing enzymes and placental fatty acid transporters were compared. Mothers with PE had higher plasma triglyceride (21%, P<0.001) and nonesterified fatty acid (50%, P<0.001) concentrations than controls. Concentrations of major n-6 and n-3 long-chain polyunsaturated fatty acids in erythrocytes were 23% to 60% lower (all P<0.005) in PE and intrauterine growth restriction mothers and offspring compared with controls. Subcutaneous adipose tissue Δ-5 and Δ-6 desaturase and very long-chain fatty acid elongase mRNA expression was lower in PE than controls (respectively, mean [SD] control 3.38 [2.96] versus PE 1.83 [1.91], P=0.030; 3.33 [2.25] versus 1.03 [0.96], P<0.001; 0.40 [0.81] versus 0.00 [0.00], P=0.038 expression relative to control gene [square root]). Low maternal and fetal long-chain polyunsaturated fatty acid concentrations in PE may be the result of decreased maternal synthesis.


Systematic analytical characterization of new psychoactive substances: A case study.
New psychoactive substances (NPS) are synthesized compounds that are not usually covered by European and/or international laws. With a slight alteration in the chemical structure of existing illegal substances registered in the European Union (EU), these NPS circumvent existing controls and are thus referred to as "legal highs". They are becoming increasingly available and can easily be purchased through both the internet and other means (smart shops). Thus, it is essential that the identification of NPS keeps up with this rapidly evolving market. In this case study, the Belgian Customs authorities apprehended a parcel, originating from China, containing two samples, declared as being "white pigments". For routine identification, the Belgian Customs Laboratory first analysed both samples by gas-chromatography mass-spectrometry and Fourier-Transform Infrared spectroscopy. The information obtained by these techniques is essential and can give an indication of the chemical structure of an unknown substance but not the complete identification of its structure. To bridge this gap, scientific and technical support is ensured by the Joint Research Centre (JRC) to the European Commission Directorate General for Taxation and Customs Unions (DG TAXUD) and the Customs Laboratory European Network (CLEN) through an Administrative Arrangement for fast recognition of NPS and identification of unknown chemicals. The samples were sent to the JRC for a complete characterization using advanced techniques and chemoinformatic tools. The aim of this study was also to encourage the development of a science-based policy driven approach on NPS. These samples were fully characterized and identified as 5F-AMB and PX-3 using (1)H and (13)C nuclear magnetic resonance (NMR), high-resolution tandem mass-spectrometry (HR-MS/MS) and Raman spectroscopy. A chemoinformatic platform was used to manage, unify analytical data from multiple techniques and instruments, and combine it with chemical and structural information.


Stability of RNA from the retina and retinal pigment epithelium in a porcine model simulating human eye bank conditions.
To assess RNA stability after death in a porcine model to simulate current human eye bank techniques. Eye bank time interval data were collected from 191 donor specimens: death to refrigeration, enucleation, and tissue processing. A control porcine eye was enucleated, retina and RPE isolated, and specimens frozen (-80 degrees C). Fourteen porcine eyes remained at room temperature for 2 hours and then cooled to 4 degrees C. Retina and RPE were isolated and frozen (-80 degrees C) at 5, 12, 24, 29, 36, 48, and 72 hours. Four globes remained in a moist chamber, five whole and five sectioned globes were immersed in RNAlater (Ambion, Austin, TX) at 5, 12, 24, or 48 hours. RNA was isolated. The 28S and 18S rRNA peaks were analyzed by electrophoresis. RT-PCR was performed on each sample. Messenger RNA for GAPDH, beta-actin, mouse rhodopsin from retina (mRHO), and RPE-65 (from RPE) were analyzed with gel electrophoresis. The average time from death to refrigeration was 4.2 hours, to enucleation 6.4 hours, and to tissue processing 10.7 hours. RT-PCR gel electrophoresis patterns from retinal tissue had bands of similar intensity at each interval from beta-actin, GAPDH, and RHO. Band patterns from RPE demonstrated decay of the RT-PCR gene products after 5 hours. This decay was delayed by at least 24 hours with the use of RNAlater. The 28S rRNA decay was similar for retina and RPE. Retinal tissue RNA can be analyzed within the time constraints of current eye bank tissue processing, whereas analysis of RPE necessitates either rapid processing or use of RNAlater. These results should aid in future studies in which eye bank tissue is used for RNA analysis.


Knowledge and practice of breast-self examination among female undergraduate students of Ahmadu Bello University Zaria, northwestern Nigeria.
Carcinoma of the breast is an important public health problem in Nigeria and studies have reported low levels of awareness and practice of breast self examination as an important method of prevention. Breast self examination is a cost-effective method of early detection of cancer of the breast especially in resource poor countries. We assessed knowledge and practice of breast-self examination (BSE) among female undergraduate students of Ahmadu Bello University Zaria, Nigeria. In this study, knowledge and practice of BSE were examined among 221 female students aged 16-28 years old studying at Ahmadu Bello University Zaria using self administered questionnaires. It was found that despite nearly three quarter of the respondents (87.7%) had heard of BSE, only 19.0% of them were performing this examination monthly. Regarding the sources of information about BSE among respondents, media was found to be most common followed by health workers accounting for 45.5% and 32.2% respectively. Regular performance of BSE was significantly correlated with duration of stay in the University (X2 = 81.9, df = 3, P 2 = 17.4, df = 2, P CONCLUSION: We observed a disparity between high levels of knowledge of BSE compared to a low level of practice. Public health education using the media could significantly reduce the knowledge-practice gap and early detection of breast lump.


[Infection of Anopheles (Kerteszia) cruzii by Plasmodium vivax and Plasmodium vivax variant VK247 in the municipalities of São Vicente and Juquitiba, São Paulo].
Sporadic cases of autochthonous malaria have been recorded in São Paulo State, located in the Southeast region of Brazil. These cases are characterized by their benign course, low parasitemia, and mild symptomatology and have been identified as vivax malaria. Little is known about the symptoms and immune response elicited in humans by the variants Plasmodium vivax VK247 and P. vivax-like human malaria parasites. These variants are transmitted by Anopheles (Kerteszia) cruzii, one of the most common species of mosquitoes in the Southeast of Brazil. The objective of the study described in this paper was to investigate infection in anophelines using ELISA immunoenzymatic assay with specific monoclonal antibodies directed against the repetitive regions of the circumsporozoite protein in classic P. vivax, P. brasilianum/P. malariae, and P. vivax VK247. Between 1991 and 1993, mosquitoes were collected in São Vicente and Juquitiba, municipalites located in a remnant of the Brazilian Atlantic forest in São Paulo State, an ecosystem rich in plants of the Bromeliaceae family. These plants function as nurseries for immature forms of anophelines of the subgenus Kerteszia. Of 1,117 An. (Ker.) cruzii captured in São Vicente, 0.179% were positive for classic P. vivax. In Juquitiba, of 1,161 An. (Ker.) cruzii, 0.086% were positive for P. vivax VK247, confirming the presence of this variant in the region. Although the infection rate is low, the high density of these mosquitoes and their voracity (they exhibit 24-h biting activity) could compensate for the low percentage of infected specimens.


Heterogeneous host susceptibility enhances prevalence of mixed-genotype micro-parasite infections.
Dose response in micro-parasite infections is usually shallower than predicted by the independent action model, which assumes that each infectious unit has a probability of infection that is independent of the presence of other infectious units. Moreover, the prevalence of mixed-genotype infections was greater than predicted by this model. No probabilistic infection model has been proposed to account for the higher prevalence of mixed-genotype infections. We use model selection within a set of four alternative models to explain high prevalence of mixed-genotype infections in combination with a shallow dose response. These models contrast dependent versus independent action of micro-parasite infectious units, and homogeneous versus heterogeneous host susceptibility. We specifically consider a situation in which genome differences between genotypes are minimal, and highly unlikely to result in genotype-genotype interactions. Data on dose response and mixed-genotype infection prevalence were collected by challenging fifth instar Spodoptera exigua larvae with two genotypes of Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV), differing only in a 100 bp PCR marker sequence. We show that an independent action model that includes heterogeneity in host susceptibility can explain both the shallow dose response and the high prevalence of mixed-genotype infections. Theoretical results indicate that variation in host susceptibility is inextricably linked to increased prevalence of mixed-genotype infections. We have shown, to our knowledge for the first time, how heterogeneity in host susceptibility affects mixed-genotype infection prevalence. No evidence was found that virions operate dependently. While it has been recognized that heterogeneity in host susceptibility must be included in models of micro-parasite transmission and epidemiology to account for dose response, here we show that heterogeneity in susceptibility is also a fundamental principle explaining patterns of pathogen genetic diversity among hosts in a population. This principle has potentially wide implications for the monitoring, modeling and management of infectious diseases.


Quantification of Aquaporin-CHIP water channel protein in microdissected renal tubules by fluorescence-based ELISA.
Several transporters have been localized along the nephron by physiological methods or immunocytochemistry. However, the actual abundance of these molecules has not been established. To accomplish this goal, we have developed a fluorescence-based ELISA method and have used it to quantitate Aquaporin-CHIP (AQP-CHIP) water channel protein in rat kidney tubules. Microdissected tubules (2 mm/sample, permeabilized with 0.5% Triton X-100) or purified AQP-CHIP standards (0-200 fmol) were utilized in a fluorescence ELISA protocol after covalent immobilization on epoxy-activated Sepharose beads. The lower limit of detection was 2.4 fmol of AQP-CHIP. Preabsorption with excess purified AQP-CHIP or use of nonimmune serum eliminated the signal. In proximal segments, the measured AQP-CHIP was linearly related to tubule length (1-10 mm). The measured AQP-CHIP was (mean +/- SE, fmol/mm): S-1 proximal, 10.8 +/- 2.1; S-2, 10.0 +/- 2.3; S-3, 21.3 +/- 3.1; type 1 thin descending limb (DTL), 12.9 +/- 4.6; type 2 DTL, 86.5 +/- 19.5; type 3 DTL, 43.0 +/- 11.2. In thin ascending limbs, thick ascending limbs, distal convoluted tubules, connecting tubules, and collecting ducts, the AQP-CHIP signal was indistinguishable from zero. Based on the unit water conductance of single CHIP molecules, our calculations show that the content of AQP-CHIP is sufficient to explain water permeability measured in isolated proximal tubules and DTL segments.


Use of honey associated with Ananas comosus (Bromelin) in the treatment of acute irritative cough.
To evaluate the immediate improvement rate of irritative cough in patients treated with the combination of Ananas comosus extract and honey (Bromelin®) compared with the use of honey alone (placebo group). Pragmatic, double-blind, randomized, parallel-group study with children aged between 2 and 15 years, with irritative cough for at least 24hours. The double-blind assessment of cough was through the number of observed coughing episodes and intensity score for a period of 10minutes of observation. The decrease of one point in the mean total score was considered as a therapeutic effect. There was a reduction in coughing episodes in both groups, as well as in the cough score after 30minutes of drug or honey administration. The change in clinical score above two points, which could indicate marked improvement, occurred in five patients in the bromelin group and only in one in the placebo group, but without significant difference. There were no adverse events. The immediate improvement rate of irritative cough was similar in patients treated with combination of Ananas comosus extract and honey (Bromelin®) compared with the use of honey alone (placebo group). It is possible that honey has a therapeutic effect on mucus and cough characteristics (Clinical Trials: NCT01356693).


Curcumin-induced mitotic spindle defect and cell cycle arrest in human bladder cancer cells occurs partly through inhibition of aurora A.
Curcumin, an active compound in turmeric and curry, has been proven to induce tumor apoptosis and inhibit tumor proliferation, invasion, angiogenesis, and metastasis via modulating numerous targets in various types of cancer cells. Aurora A is a mitosis-related serine-threonine kinase and plays important roles in diverse human cancers. However, the effect of curcumin on Aurora A has not been reported. In this study, Aurora A promoter activity and mRNA expression were inhibited in curcumin-treated human bladder cancer T24 cells, suggesting that Aurora A is regulated at the transcription level. We also found that curcumin preferentially inhibited the growth of T24 cells, which show a higher proliferation rate, invasion activity, and expression level of Aurora A compared with that of human immortalized uroepithelial E7cells. Furthermore, inhibition of phosphorylation of Aurora A and its downstream target histone H3 accompanied by the formation of monopolar spindle, induction of G(2)/M phase arrest, and reduction in cell division in response to curcumin were detected in T24 cells. These curcumin-induced phenomena were similar to those using Aurora A small interfering RNA and were attenuated by ectopic expression of Aurora A. Therefore, the antitumor mechanism of curcumin is Aurora A-related, which further supports the application of curcumin in treatments of human cancers.


Gender comparisons of anthropometric characteristics of young sprint swimmers.
The purpose of this study was to compare the body composition, body build, and anthropometric characteristics of boy and girl sprint swimmers. Two groups (boys, n = 38 and girls, n = 31) of sprint swimmers (mean age ± SD = 11.03 ± 2.29 and 10.45 ± 2.29 years, respectively) volunteered for this study. The subjects were members of local swimming clubs who competed in sprint swimming events (≤ 200 m). Gender comparisons were made for age, body weight (BW), height (HT), fat-free weight (FFW), percent body fat (%fat), endomorphic rating, mesomorphic rating, ectomorphic rating, sum of 12 diameters, sum of 11 circumferences, biacromial diameter/biiliac diameter, and FFW/HT. The results of the independent t-tests indicated that the only mean differences between the boy and girl sprint swimmers were for % fat (boys = 9.40 ± 5.35% fat; girls = 12.73 ± 6.19% fat) and endomorphic rating (boys = 2.87 ± 0.96; girls = 4.29 ± 1.22). For the current age group of sprint swimmers the only gender differences were for measures associated with body fatness, and there were no differences for body build measures associated with musculoskeletal size, muscularity, skeletal size, total body mass, or body breadth dimensions. These findings suggest that the swimming performance for girls may be improved through training programs designed to reduce body fatness.


Embryo toxicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin to the wood duck (Aix sponsa).
We examined the sensitivity of the wood duck (Aix sponsa) embryo to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) by injecting the toxicant into their eggs. Six groups of wood duck eggs (n = 35 to 211 per trial) were injected with 0 to 4600 pg TCDD/g egg between 2003 and 2005. Injections were made into yolk prior to incubation, and eggs were subsequently incubated and assessed weekly for mortality. Significant TCDD-induced mortality was not observed through day 25 (90% of incubation). Liver, heart, eye, and brain histology were generally unremarkable. Hepatic ethoxyresorufin-O-deethylase activity, a biomarker of dioxin-like compound exposure, was induced by 12-fold in the 4600 pg/g treatment relative to controls. The median lethal dose for chicken (Gallus domesticus) eggs we dosed identically to wood duck eggs was about 100 pg/g, similar to other assessments of chickens. Among dioxin-like compound embryo lethality data for 15 avian genera, the wood duck 4600 pg/g no-observed-effect level ranks near the middle. Because no higher doses were tested, wood ducks may be like other waterfowl (order Anseriformes), which are comparatively tolerant to embryo mortality from polychlorinated dibenzo-p-dioxins and dibenzofurans when exposed by egg injection.


Engineering Biomimetic Nanofiber Microspheres with Tailored Size, Predesigned Structure, and Desired Composition via Gas Bubble-Mediated Coaxial Electrospray.
Minimally invasive therapies avoiding surgical complexities evoke great interest in developing injectable biomedical devices. Herein, a versatile approach is reported for engineering injectable and biomimetic nanofiber microspheres (NMs) with tunable sizes, predesigned structures, and desired compositions via gas bubble-mediated coaxial electrospraying. The sizes and structures of NMs are controlled by adjusting processing parameters including air flow rate, applied voltage, distance, and spinneret configuration in the coaxial setup. Importantly, unlike the self-assembly method, this technique can be used to fabricate NMs from any material feasible for electrospinning or other nanofiber fabrication techniques. To demonstrate the versatility, open porous NMs are successfully fabricated that consist of various short nanofibers made of poly(ε-caprolactone), poly(lactic-co-glycolic acid), gelatin, methacrylated gelatin, bioglass, and magneto-responsive polymer composites. Open porous NMs support human neural progenitor cell growth in 3D with a larger number and more neurites than nonporous NMs. Additionally, highly open porous NMs show faster cell infiltration and host tissue integration than nonporous NMs after subcutaneous injection to rats. Such a novel class of NMs holds great potential for many biomedical applications such as tissue filling, cell and drug delivery, and minimally invasive tissue regeneration.


The distribution of fruit and seed toxicity during development for eleven neotropical trees and vines in Central Panama.
Secondary compounds in fruit mediate interactions with natural enemies and seed dispersers, influencing plant survival and species distributions. The functions of secondary metabolites in plant defenses have been well-studied in green tissues, but not in reproductive structures of plants. In this study, the distribution of toxicity within plants was quantified and its influence on seed survival was determined in Central Panama. To investigate patterns of allocation to chemical defenses and shifts in allocation with fruit development, I quantified variation in toxicity between immature and mature fruit and between the seed and pericarp for eleven species. Toxicity of seed and pericarp was compared to leaf toxicity for five species. Toxicity was measured as reduced hyphal growth of two fungal pathogens, Phoma sp. and Fusarium sp., and reduced survivorship of brine shrimp, Artemia franciscana, across a range of concentrations of crude extract. I used these measures of potential toxicity against generalist natural enemies to examine the effect of fruit toxicity on reductions of fruit development and seed survival by vertebrates, invertebrates, and pathogens measured for seven species in a natural enemy removal experiment. The seed or pericarp of all vertebrate- and wind-dispersed species reduced Artemia survivorship and hyphal growth of Fusarium during the immature and mature stages. Only mature fruit of two vertebrate-dispersed species reduced hyphal growth of Phoma. Predispersal seed survival increased with toxicity of immature fruit to Artemia during germination and decreased with toxicity to fungi during fruit development. This study suggests that fruit toxicity against generalist natural enemies may be common in Central Panama. These results support the hypothesis that secondary metabolites in fruit have adaptive value and are important in the evolution of fruit-frugivore interactions.


Nurse-patient interaction and decision-making in care: patient involvement in community nursing.
This paper reports a study to identify the extent to which community nurses incorporate patient involvement in decision-making into their everyday clinical practice. The complexity underlying the concept of patient participation is not always recognized. It links the notions of 'passive patient' and 'active participant', hence its usage along with those of similar terms such as 'partnership', 'involvement' and 'collaboration' is often ambiguous. The literature on this subject is fairly extensive and often contradictory; it encompasses the participation of potential as well as actual patients. Hierarchical structures have been proposed to offer an understanding of the current nature of patient participation, and these have sometimes linked the level of participation to the quality of the nurse-patient relationship. Much work has been done in the area of cancer care, where it has been noted that the role of 'passive patient' may in itself be a deliberate choice. The approach was ethnographic and data were collected during 2000 and 2001 by participant observations with 22 nurses and 107 patients; 137 interactions were observed in all. Detailed field notes were kept and then carefully interpreted. The approach to rigour was primarily concerned with ensuring credibility. The extent to which nurses involved patients in making decisions about their care, and the manner in which this was achieved, varied considerably. Collectively, the behaviour of participants is presented as the 'involving-non-involving continuum', with five typologies of behaviour: 'completely involving'; 'partially involving'; 'forced involving'; 'covert non-involving' and 'overt non-involving'. The findings highlight that caring for people in their own home does not necessarily mean that patients are involved in care and treatment decisions. This observation study of the extent to which community nurses involved patients in decision-making in nursing care offers some useful insights into one aspect of nursing work. The implications of the study are far-reaching. The findings suggest that it could be important for nurses and their managers to devote more time to relationship-building in nursing practice. They also offer a perspective--the involving-non-involving continuum--that could usefully be incorporated into nurse education at both pre- and postregistration levels.


Value of high-sensitivity C-reactive protein in low risk chest pain observation unit patients.
High-sensitivity C-reactive protein (hs-CRP) rises with cardiac injury/ischemia. We evaluated its efficacy in aiding in the identification of an acute coronary syndrome (ACS) in patients (pts) admitted to the chest pain unit (CPU) for possible ACS. Retrospective study of all patients admitted to the CPU with chest pain who underwent hs-CRP testing as part of their CPU evaluation from January 2004 to October 2008. Patients were low risk for ACS (compatible symptoms, nondiagnostic initial ECG, and negative cTnI). ACS was diagnosed by positive functional study, cardiac catheterization, or cardiac event during 30-day follow-up. Positive hs-CRP was defined based on local laboratory levels (>1.0 mg/l or >3.0 mg/l), and population-based and prior study values >2.0 mg/l. Chi-square analysis was performed, and odds ratios (OR) are presented. Multivariate analysis was done to determine whether hs-CRP was independently associated with the diagnosis of ACS. Cardiac risk factors, demographics, and diagnosis of ACS were included in the model. Medians with IQR are presented for continuous data. Ninety-five percent confidence intervals are presented where applicable. A total of 958 patients had hs-CRP testing as part of their CPEU evaluation. Excluded from the analysis were 39 patients lost to follow-up. The final cohort comprised 478 (52%) women and 441 (48%) men with a median age of 56 (IQR 48-64). ACS was diagnosed in 128 (13.4%). The median cohort hs-CRP value was 2.2 mg/l (IQR 0.7, 5.8) and 2.3 mg/l (IQR 0.6, 5.9) in those with and without ACS, respectively. In the multivariate analysis hs-CRP was not independently associated with the diagnosis of ACS (0.99; 95% CI 0.98 - 1.01). In large patient cohort managed in a single-center CPU, measurement of hs-CRP did not enhance the diagnostic accuracy for ACS. Routine hs-CRP as a diagnostic tool should not be recommended in the CPU setting.


Replication Fork Protection Factors Controlling R-Loop Bypass and Suppression.
Replication-transcription conflicts have been a well-studied source of genome instability for many years and have frequently been linked to defects in RNA processing. However, recent characterization of replication fork-associated proteins has revealed that defects in fork protection can directly or indirectly stabilize R-loop structures in the genome and promote transcription-replication conflicts that lead to genome instability. Defects in essential DNA replication-associated activities like topoisomerase, or the minichromosome maintenance (MCM) helicase complex, as well as fork-associated protection factors like the Fanconi anemia pathway, both appear to mitigate transcription-replication conflicts. Here, we will highlight recent advances that support the concept that normal and robust replisome function itself is a key component of mitigating R-loop coupled genome instability.


Versatility and connectivity efficiency of bipartite transcription networks.
The modulation of promoter activity by DNA-binding transcription regulators forms a bipartite network between the regulators and genes, in which a smaller number of regulators control a much lager number of genes. To facilitate representation of gene expression data with the simplest possible network structure, we have characterized the ability of bipartite networks to describe data. This has led to the classification of two types of bipartite networks, versatile and nonversatile. Versatile networks can describe any data of the same rank, and are indistinguishable from one another. Nonversatile networks require constraints to be present in data they describe, which may be used to distinguish between different network topologies. By quantifying the ability of bipartite networks to represent data we were able to define connectivity efficiency, which is a measure of how economic the use of connections is within a network with respect to data representation and generation. We postulated that it may be desirable for an organism to maximize its gene expression range per network edge, since development of a regulatory connection may have some evolutionary cost. We found that the transcriptional regulatory networks of both Saccharomyces cerevisiae and Escherichia coli lie close to their respective connectivity efficiency maxima, suggesting that connectivity efficiency may have some evolutionary influence.


Fast inhibition of glutamate-activated currents by caffeine.
Caffeine stimulates calcium-induced calcium release (CICR) in many cell types. In neurons, caffeine stimulates CICR presynaptically and thus modulates neurotransmitter release. Using the whole-cell patch-clamp technique we found that caffeine (20 mM) reversibly increased the frequency and decreased the amplitude of miniature excitatory postsynaptic currents (mEPSCs) in neocortical neurons. The increase in mEPSC frequency is consistent with a presynaptic mechanism. Caffeine also reduced exogenously applied glutamate-activated currents, confirming a separate postsynaptic action. This inhibition developed in tens of milliseconds, consistent with block of channel currents. Caffeine (20 mM) did not reduce currents activated by exogenous NMDA, indicating that caffeine block is specific to non-NMDA type glutamate receptors. Caffeine-induced inhibition of mEPSC amplitude occurs through postsynaptic block of non-NMDA type ionotropic glutamate receptors. Caffeine thus has both pre and postsynaptic sites of action at excitatory synapses.


The malaria transition on the Arabian Peninsula: progress toward a malaria-free region between 1960-2010.
The transmission of malaria across the Arabian Peninsula is governed by the diversity of dominant vectors and extreme aridity. It is likely that where malaria transmission was historically possible it was intense and led to a high disease burden. Here, we review the speed of elimination, approaches taken, define the shrinking map of risk since 1960 and discuss the threats posed to a malaria-free Arabian Peninsula using the archive material, case data and published works. From as early as the 1940s, attempts were made to eliminate malaria on the peninsula but were met with varying degrees of success through to the 1970s; however, these did result in a shrinking of the margins of malaria transmission across the peninsula. Epidemics in the 1990s galvanised national malaria control programmes to reinvigorate control efforts. Before the launch of the recent global ambition for malaria eradication, countries on the Arabian Peninsula launched a collaborative malaria-free initiative in 2005. This initiative led a further shrinking of the malaria risk map and today locally acquired clinical cases of malaria are reported only in Saudi Arabia and Yemen, with the latter contributing to over 98% of the clinical burden.


Negative pressure wound therapy with instillation: past, present, and future.
Negative pressure wound therapy with instillation (NPWTi) is a novel treatment option that provides the combination of negative pressure with intermittent instillation of a solution. Standard Negative Pressure Wound Therapy (NPWT) is an established adjunctive treatment option that offers the ability to promote granulation tissue in wounds. However, there is limited evidence for its utility in the environment of active or senescent infection. Wounds that are acutely infected or that contain deleterious biofilm are a challenging problem, which require an intensive multimodal approach including antibiosis, surgical intervention, and local wound care. Adjunctive application of NPWTi can potentially expedite clearance of infection and wound closure. Although this technology has been commercially available for over a decade, its adoption has been limited. Recently, there has been a resurgence of interest in this therapy with emerging evidence from animal models as well as human clinical studies. There are remaining questions regarding NPWTi including the selection of the optimal instillation solution and device settings. This article discusses the past development, current knowledge, and future direction of NPWTi.


PML-RARalpha/RXR Alters the Epigenetic Landscape in Acute Promyelocytic Leukemia.
Many different molecular mechanisms have been associated with PML-RARalpha-dependent transformation of hematopoietic progenitors. Here, we identified high confidence PML-RARalpha binding sites in an acute promyelocytic leukemia (APL) cell line and in two APL primary blasts. We found colocalization of PML-RARalpha with RXR to the vast majority of these binding regions. Genome-wide epigenetic studies revealed that treatment with pharmacological doses of all-trans retinoic acid induces changes in H3 acetylation, but not H3K27me3, H3K9me3, or DNA methylation at the PML-RARalpha/RXR binding sites or at nearby target genes. Our results suggest that PML-RARalpha/RXR functions as a local chromatin modulator and that specific recruitment of histone deacetylase activities to genes important for hematopoietic differentiation, RAR signaling, and epigenetic control is crucial to its transforming potential.


Mild measles and secondary vaccine failure during a sustained outbreak in a highly vaccinated population.
A prolonged school-based outbreak of measles provided an opportunity to study "vaccine-modified" mild measles and secondary vaccine failure. Thirty-six (97%) of 37 unvaccinated patients had rash illnesses that met the Centers for Disease Control clinical case definition of measles, but 29 (15%) of 198 vaccinated patients did not, primarily because of low-grade or absent fever. Of 122 patients with seroconfirmed measles, 10 patients (all previously vaccinated) had no detectable measles-specific IgM and significantly milder illness than either vaccinated or unvaccinated patients with IgM-positive serum. Of 108 vaccinated patients with seroconfirmed measles, 17 patients (16%) had IgM-negative serology or rash illnesses that failed to meet the clinical case definition; their mean age (13 years), age at the time of vaccination, and time since vaccination did not differ from those of other vaccinated patients. The occurrence of secondary vaccine failure and vaccine-modified measles does not appear to be a major impediment to measles control in the United States but may lead to underreporting of measles cases and result in overestimation of vaccine efficacy in highly vaccinated populations.


Genetic red flags: clues to thinking genetically in primary care practice.
This article presents an approach to "thinking genetically" in primary care. Busy practitioners often lack the time to consider thoroughly whether their patients have an underlying genetic diagnosis. To assist the primary care clinician, a working group of the Genetics in Primary Care Faculty Development Initiative developed a simple mnemonic, Family GENES, that alerts the clinician to consider genetic causes in the differential diagnosis. In addition to family history, the red flags include Groups of anomalies, Early or Extreme presentations of common diseases, Neurodevelopmental or Neurodegenerative conditions, Exceptional or unusual pathology, and Surprising laboratory values. This article discusses the components of the mnemonic, provides examples, and gives guidelines to appropriate actions once the possibility of a genetic diagnosis has been raised.


Quantitative relationships between lacZ mutant frequency and DNA adduct frequency in Muta™Mouse tissues and cultured cells exposed to 3-nitrobenzanthrone.
The frequency of stable DNA adducts in a target tissue can be used to assess biologically effective dose; however, the utility of the metric in a risk assessment context depends on the likelihood that the DNA damage will be manifested as mutation. Previously, we employed the Muta™Mouse system to examine the induction of lacZ mutants and DNA adducts following exposure to the well-studied mutagenic carcinogen 3-nitrobenzanthrone (3-NBA). In this follow-up work, we examined the empirical relationships between total adduct frequency and mutant frequency (MF) in tissues and cultured cells following acute 3-NBA exposure. The results show a significant induction of DNA damage and lacZ mutants in liver, colon and bone marrow, as well as FE1 pulmonary epithelial cells. In contrast, lung and small intestine samples had low, but significantly elevated adduct levels, with no significant increases in lacZ MF. Additional analyses showed a significant relationship between the mutagenic efficiency of total adducts, measured as the slope of the relationships between MF and total adduct frequency, and tissue-specific mitotic index (MI). The lack of mutation response in lung, in contrast to the high in vitro MF in FE-1 lung cells, is likely related to the 100-fold difference in MI. The lack of small intestine mutagenic response may be related to limited metabolic capacity, differences in DNA repair, and /or chemically induced apoptosis that has been observed for other potent mutagens. The results indicate that interpretation of adduct frequency values in a risk assessment context can be improved by considering the MI of the target tissue; however, more generalised interpretation is hampered by tissue-specific variations in metabolic capacity and damage processing. The work provides a proof of principle regarding the use of the Muta™Mouse system to critically examine the health risks associated with tissue-specific adduct loads.


For faculty, by faculty: a case study of learning to teach online.
The shortage of nursing faculty and the need for MSN-prepared faculty to have access to doctoral education and remain in their teaching roles has resulted in a growing number of nurse education programs moving online. A better understanding of how best to support faculty during this transition is needed. This case study describes the experiences of faculty at one institution as they participated in a grassroots effort to learn about online teaching. Six themes related to the faculty development experience were identified: 1) plugging in; 2) peer sharing, modeling and community building; 3) multidimensional learning; 4) role-shifting and meta-learning; 5) paradigm shifting; and 6) sustaining momentum. Findings are connected to recommendations related to how best to prepare faculty to ensure that quality nursing education continues.


Acute asthma epidemics, weather and pollen in England, 1987-1994.
Recent epidemics of acute asthma have caused speculation that, if their causes were known, early warnings might be feasible. In particular, some epidemics seemed to be associated with thunderstorms. We wondered what risk factors predicting epidemics could be identified. Daily asthma admissions counts during 1987-1994, for two age groups (0-14 yrs and > or = 15 yrs), were measured using the Hospital Episodes System (HES). Epidemics were defined as combinations of date, age group and English Regional Health Authority (RHA) with exceptionally high asthma admission counts compared to the predictions of a log-linear autoregression model. They were compared with control days 1 week before and afterwards, regarding seven meteorological variables and 5 day average pollen counts for four species. Fifty six asthma epidemics were identified. The mean density of sferics (lightning flashes), temperature and rainfall on epidemic days were greater than those on control days. High sferics densities were overrepresented in epidemics. Simultaneously high sferics and grass pollen further increased the probability of an epidemic, but only to 15% (95% confidence interval 2-45%). Two thirds of epidemics were not preceded by thunderstorms. Thunderstorms and high grass pollen levels precede asthma epidemics more often than expected by chance. However, most epidemics are not associated with thunderstorms or unusual weather conditions, and most thunderstorms, even following high grass pollen levels, do not precede epidemics. An early warning system based on the indicators examined here would, therefore, detect few epidemics and generate an unacceptably high rate of false alarms.


Intralimb coordination as a sensitive indicator of motor-control impairment after spinal cord injury.
Recovery of walking function after neurotrauma, e.g., after spinal cord injury, is routinely captured using standardized walking outcome measures of time and distance. However, these measures do not provide information on possible underlying mechanisms of recovery, nor do they tell anything about the quality of gait. Subjects with an incomplete spinal cord injury are a very heterogeneous group of people with a wide range of functional impairments. A stratification of these subjects would allow increasing sensitivity for hypothesis testing and a more targeted treatment strategy. The gait of incomplete spinal cord injured subjects was compared to healthy control subjects by analyzing kinematic data obtained by a 3-D motion capture system. Hip-knee angle-angle plots (cyclograms) informed on the qualitative aspect of gait and the intralimb coordination. Features of the cyclogram, e.g., shape of the cyclogram, cycle-to-cycle consistency and its modulation due to changes in walking speed were discerned and used to stratify spinal cord injured subjects. Spinal cord injured subjects were unable to modulate their cyclogram configuration when increasing speed from slow to preferred. Their gait quality remained clearly aberrant and showed even higher deviations from normal when walking at preferred speed. Qualitative categorization of spinal cord injured subjects based on their intralimb coordination was complemented by quantitative measures of cyclogram shape comparison. Spinal cord injured subjects showed distinct distortions of intralimb coordination as well as limited modulation to changes in walking speed. The specific changes of the cyclograms revealed complementary insight in the disturbance of lower-limb control in addition to measures of time and distance and may be a useful tool for patient categorization and stratification prior to clinical trial inclusion.


Sand dune ridge alignment effects on surface BRF over the Libya-4 CEOS calibration site.
The Libya-4 desert area, located in the Great Sand Sea, is one of the most important bright desert CEOS pseudo-invariant calibration sites by its size and radiometric stability. This site is intensively used for radiometer drift monitoring, sensor intercalibration and as an absolute calibration reference based on simulated radiances traceable to the SI standard. The Libya-4 morphology is composed of oriented sand dunes shaped by dominant winds. The effects of sand dune spatial organization on the surface bidirectional reflectance factor is analyzed in this paper using Raytran, a 3D radiative transfer model. The topography is characterized with the 30 m resolution ASTER digital elevation model. Four different regions-of-interest sizes, ranging from 10 km up to 100 km, are analyzed. Results show that sand dunes generate more backscattering than forward scattering at the surface. The mean surface reflectance averaged over different viewing and illumination angles is pretty much independent of the size of the selected area, though the standard deviation differs. Sun azimuth position has an effect on the surface reflectance field, which is more pronounced for high Sun zenith angles. Such 3D azimuthal effects should be taken into account to decrease the simulated radiance uncertainty over Libya-4 below 3% for wavelengths larger than 600 nm.


Increased alcohol use during the COVID-19 pandemic: The effect of mental health and age in a cross-sectional sample of social media users in the U.S.
The COVID-19 pandemic has triggered a public health crisis of unprecedented scale. Increased alcohol use has been extensively documented during other crises, particularly among persons with anxiety and depression. Despite COVID-19's differential impact by age, the association of age, mental health and alcohol use during the pandemic has not been explored. This study aimed to examine whether age modified the association of anxiety and depressive symptoms with alcohol use during the COVID-19 pandemic. Two online surveys were administered to U.S. adult social media users in March and April 2020. Generalized linear models were conducted in 2020 among 5850 respondents (52.9% female; 22.0% aged 18-39 years, 47.0% aged 40-59 years, and 31.0% aged ≥60 years) to examine if age modified the association of anxiety and depression symptomatology and alcohol use. Overall, 29% of respondents reported increased alcohol use. Adjusted odds ratios of reporting increased alcohol use were 1.41 (95% CI = 1.20-1.66) among respondents with anxiety symptoms and 1.64 (95% CI = 1.21-2.23) among those with depressive symptoms compared to those without such symptoms. Whereas respondents aged 18-39 years had the highest probability of reporting increased alcohol use, the probability of older persons (40-59 and ≥60 years) reporting increased drinking was much greater among those with symptoms of anxiety and depression, compared to those without symptoms. These findings warrant age-differentiated public health messaging on the risks of excessive alcohol use and scale-up of substance use services for middle-aged and older adults with symptoms of depression and anxiety.


Plasma noradrenaline concentrations and haemodynamics in the early stage of essential hypertension.
1. Plasma noradrenaline concentrations and haemodynamic status were simultaneously studied in young patients with uncomplicated essential hypertension and in age-matched normal controls. 2. Resting plasma noradrenaline in the controls tended to increase slightly, but progressively, with age. The hypertensive subjects had significantly higher plasma noradrenaline concentrations than those in the controls, but these values did not show any age-related variation. The response of plasma noradrenaline to the standing position tended to increase with age in the controls, whereas plasma noradrenaline in the hypertensive subjects showed a wide range of responses without any fixed relationship with age. 3. The cardiac index was significantly greater in the labile hypertensive subjects than in the controls, whereas total peripheral resistance was significantly greater in the sustained hypertensive subjects than in the labile patients and in the controls. Mean arterial pressure in these patients was closely related with the value of total peripheral resistance rather than with the cardiac index. 4. Of the patients with raised plasma noradrenaline 80% showed significantly increased values of either total peripheral resistance or cardiac index. Plasma noradrenaline was correlated significantly to total peripheral resistance, and marginally to mean arterial pressure. 5. These findings support the view that sympathetic nervous overactivity is an important factor underlying the haemodynamic findings in these patients.


Novel POLG1 mutations in a patient with adult-onset progressive external ophthalmoplegia and encephalopathy.
Mutations in POLG1 are an important cause of human mitochondrial disease. We describe a woman who presented with bilateral ptosis and external ophthalmoplegia at 64 years of age. Neurological examination revealed symptoms of diffuse encephalopathy. The symptoms were progressive and at 67 years she was severely cognitively impaired, had severe bilateral ptosis and complete external ophthalmoplegia. Frequent cytochrome c oxidase-negative fibres were detected in muscle. Electrophysiological examination revealed myopathic changes and axonal neuropathy. Standard laboratory tests were normal. Brain CT showed general, moderate cortical atrophy. Molecular analysis of muscle DNA revealed multiple mitochondrial DNA deletions. Sequencing of the entire POLG1 gene revealed two changes c.2993C>T (p.998S>L) and c.3550G>C (p.1184D>H). Both mutations are previously unreported and confirmed to be compound heterozygous. Late-onset progressive external ophthalmoplegia with severe encephalopathy is an unusual combination in patients with POLG1 mutations. POLG-associated disease should be considered in any patient with unexplained or unusual neurological features.


Effect of various warm-up devices on bat velocity of intercollegiate baseball players.
A variety of warm-up devices are available to baseball players to use before their game at-bat. Past baseball research evaluating warm-up devices indicates that implements that are ±12% of standard game bat weight produce the greatest bat velocities for high school and intercollegiate players. The purpose of this study was to examine the effect of various warm-up devices on bat velocity (BV) of intercollegiate baseball players. Twenty-two Division I intercollegiate baseball players (age = 20.0 ± 1.5 years, height = 182.6 ± 8.3 cm, body mass = 91.4 ± 11.4 kg, lean body mass = 78.8 ± 8.9 kg, % body fat = 13.6 ± 3.8) participated in a warm-up with 1 of 10 weighted devices on separate days. Each of the 10 testing sessions consisted of a standardized warm-up, 3 dry swings as hard as possible with the assigned warm-up device, 2 comfortable dry swings with a standard game baseball bat followed by 3 game swings (20-second rest between swings) while hitting a baseball off of a batting tee with the same standard game baseball bat. Results indicated that there were no statistically significant differences in BV after using any of the 10 warm-up devices. For male intercollegiate baseball players, results indicate that warm-up devices varying from 623.7 to 2,721.5 g (22-96 oz.) did not change mean BV of a standard game baseball bat, suggesting that intercollegiate players can use any of the 10 warm-up devices in the on-deck circle and maintain their BV. Therefore, personal preference as to which warm-up implement to use in the on-deck circle is advised.


Transparent, flexible, and stretchable WS2 based humidity sensors for electronic skin.
Skin-mountable chemical sensors using flexible chemically sensitive nanomaterials are of great interest for electronic skin (e-skin) application. To build these sensors, the emerging atomically thin two-dimensional (2D) layered semiconductors could be a good material candidate. Herein, we show that a large-area WS2 film synthesized by sulfurization of a tungsten film exhibits high humidity sensing performance both in natural flat and high mechanical flexible states (bending curvature down to 5 mm). The conductivity of as-synthesized WS2 increases sensitively over a wide relative humidity range (up to 90%) with fast response and recovery times in a few seconds. By using graphene as electrodes and thin polydimethylsiloxane (PDMS) as substrate, a transparent, flexible, and stretchable humidity sensor was fabricated. This senor can be well laminated onto skin and shows stable water moisture sensing behaviors in the undeformed relaxed state as well as under compressive and tensile loadings. Furthermore, its high sensing performance enables real-time monitoring of human breath, indicating a potential mask-free breath monitoring for healthcare application. We believe that such a skin-activity compatible WS2 humidity sensor may shed light on developing low power consumption wearable chemical sensors based on 2D semiconductors.


Generation of Evc2/Limbin global and conditional KO mice and its roles during mineralized tissue formation.
Ellis-van Creveld (EvC) syndrome (OMIM 225500) is an autosomal recessive disease characterized with chondrodysplastic dwarfism in association with abnormalities in oral cavity. Ciliary proteins EVC and EVC2 have been identified as causative genes and they play an important role on Hedgehog signal transduction. We have also identified a causative gene LIMBIN for bovine chondrodysplastic dwarfism (bcd) that is later identified as the bovine ortholog of EVC2. Here, we report generation of conventional and conditional mutant Evc2/Limbin alleles that mimics mutations found in EvC patients and bcd cattle. Resulted homozygous mice showed no ciliary localization of EVC2 and EVC and displayed reduced Hedgehog signaling activity in association with skeletal and oral defects similar to the EvC patients. Cartilage-specific disruption of Evc2/Limbin resulted in similar but milder skeletal defects, whereas osteoblast-specific disruption did not cause overt changes in skeletal system. Neural crest-specific disruption of Evc2/Limbin resulted in defective incisor growth similar to that seen in conventional knockouts; however, differentiation of amelobolasts was relatively normal in the conditional knockouts. These results showcased functions of EVC2/LIMBIN during formation of mineralized tissues. Availability of the conditional allele for this gene should facilitate further detailed analyses of the role of EVC2/LIMBIN in pathogenesis of EvC syndrome. genesis 53:612-626, 2015. © 2015 Wiley Periodicals, Inc.


Appearance of a substance in acute phase serum which elicits Cx-protein responses.
Seromucoid was isolated from serum of rabbits during the acute inflammatory period following subcutaneous injection of an irritant and at times after whole body irradiation. It produced a Cxprotein response on intravenous injection into normal rabbits, an anamnestic response in immunized rabbits, leucocyte changes characteristic of inflammation, and marked toxicity in rabbits with reticuloendothelial system blockade. Normal seromucoid did not possess these properties.


Light scattering and absorption by randomly-oriented cylinders: dependence on aspect ratio for refractive indices applicable for marine particles.
Typically, explanation/interpretation of observed light scattering and absorption properties of marine particles is based on assuming a spherical shape and homogeneous composition. We examine the influence of shape and homogeneity by comparing the optics of randomly-oriented cylindrically-shaped particles with those of equal-volume spheres, in particular the influence of aspect ratio (AR=length/diameter) on extinction and backscattering. Our principal finding is that the when AR>~3-5 and the diameter is of the order of the wavelength, the extinction efficiency and the backscattering probability are close to those of an infinite cylinder. In addition, we show the spherical-based interpretation of extinction and absorption can lead to large error in predicted backscattering.


Surfactant for pulmonary haemorrhage in neonates.
In the 1960s and 1970s, pulmonary haemorrhage (PH) occurred mainly in full-term infants with pre-existing illness with an incidence of 1.3 per 1000 live births. Risk factors for PH included severity of illness, intrauterine growth restriction, patent ductus arteriosus (PDA), coagulopathy and the need for assisted ventilation. Presently, PH occurs in 3% to 5% of preterm ventilated infants with severe respiratory distress syndrome (RDS) who often have a PDA and have received surfactant. The cause of PH is thought to be due to rapid lowering of intrapulmonary pressure, which facilitates left to right shunting across a PDA and an increase in pulmonary blood flow. Retrospective case reports and one prospective uncontrolled study have shown promising results for surfactant in treating PH. To evaluate the effect of surfactant treatment compared to placebo or no intervention on mortality and morbidities in neonates with PH. For this update The Cochrane Library, Issue 2, 2012; MEDLINE; EMBASE; CINAHL; Clinicaltrials.gov; Controlled-trials.com; proceedings (2000 to 2011) of the Annual Meetings of the Pediatric Academic Societies (Abstracts2View) and Web of Science were searched on 8 February 2012. Randomised or quasi-randomised controlled trials that evaluated the effect of surfactant in the treatment of PH in intubated term or preterm (< 37 weeks) neonates with PH. Infants were included up to 44 weeks' postmenstrual age. The interventions studied were intratracheal instillation of surfactant (natural or synthetic, regardless of dose) versus placebo or no intervention. If studies were identified by the literature search, the planned analyses included risk ratio, risk difference, number needed to treat to benefit or to harm for dichotomous outcomes, and mean difference for continuous outcomes, with their 95% confidence intervals. A fixed-effect model would be used for meta-analyses. The risk of bias for included trials would be assessed. Heterogeneity tests, including the I(2) statistic, would be performed to assess the appropriateness of pooling the data and the results would be reported. No trials were identified. No randomised or quasi-randomised trials that evaluated the effect of surfactant in PH were identified. Therefore, no conclusions from such trials can be drawn. In view of the promising results from studies with less strict study designs than a randomised controlled trial, there is reason to conduct further trials of surfactant for the treatment of PH in neonates.


Urine-concentrating mechanism in the inner medulla: function of the thin limbs of the loops of Henle.
The ability of mammals to produce urine hyperosmotic to plasma requires the generation of a gradient of increasing osmolality along the medulla from the corticomedullary junction to the papilla tip. Countercurrent multiplication apparently establishes this gradient in the outer medulla, where there is substantial transepithelial reabsorption of NaCl from the water-impermeable thick ascending limbs of the loops of Henle. However, this process does not establish the much steeper osmotic gradient in the inner medulla, where there are no thick ascending limbs of the loops of Henle and the water-impermeable ascending thin limbs lack active transepithelial transport of NaCl or any other solute. The mechanism generating the osmotic gradient in the inner medulla remains an unsolved mystery, although it is generally considered to involve countercurrent flows in the tubules and vessels. A possible role for the three-dimensional interactions between these inner medullary tubules and vessels in the concentrating process is suggested by creation of physiologic models that depict the three-dimensional relationships of tubules and vessels and their solute and water permeabilities in rat kidneys and by creation of mathematical models based on biologic phenomena. The current mathematical model, which incorporates experimentally determined or estimated solute and water flows through clearly defined tubular and interstitial compartments, predicts a urine osmolality in good agreement with that observed in moderately antidiuretic rats. The current model provides substantially better predictions than previous models; however, the current model still fails to predict urine osmolalities of maximally concentrating rats.


Socioeconomic inequalities in physiological risk biomarkers and the role of lifestyles among Russians aged 35-69 years.
Socioeconomic inequalities in cardiovascular (CVD) health outcomes are well documented. While Russia has one of the highest levels of CVD mortality in the world, the literature on contemporary socio-economic inequalities in biomarker CVD risk factors is sparse. This paper aims to assess the extent and the direction of SEP inequalities in established physiological CVD risk biomarkers, and to explore the role of lifestyle factors in explaining SEP inequalities in physiological CVD risk biomarkers. We used cross-sectional data from a general population-based survey of Russians aged 35-69 years living in two cities (n = 4540, Know Your Heart study 2015-18). Logistic models were used to assess the associations between raised physiological risk biomarkers levels (blood pressure levels, cholesterol levels, triglycerides, HbA1C, and C-reactive protein) and socioeconomic position (SEP) (education and household financial constraints) adjusting for age, obesity, smoking, alcohol and health-care seeking behavior. High education was negatively associated with a raised risk of blood pressure (systolic and diastolic) and C-reactive protein for both men and women. High education was positively associated with total cholesterol, with higher HDL levels among women, and with low triglycerides and HbA1c levels among men. For the remaining risk biomarkers, we found little statistical support for SEP inequalities. Adjustment for lifestyle factors, and particularly BMI and waist-hip ratio, led to a reduction in the observed SEP inequalities in raised biomarkers risk levels, especially among women. High financial constraints were weakly associated with high risk biomarkers levels, except for strong evidence for an association with C-reactive protein (men). Notable differences in risk biomarkers inequalities were observed according to the SEP measure employed. Clear educational inequalities in raised physiological risk biomarkers levels, particularly in blood pressure and C-reactive protein were seen in Russia and are partly explained by lifestyle factors, particularly obesity among women. These findings provide evidence-based information on the need for tackling health inequalities in the Russian population, which may help to further contribute to CVD mortality decline.


Immune response to melanoma extracts in three melanoma-prone families.
Immune reactivity to melanoma extracts was measured by the leukocyte adherence inhibition (LAI) test in 40 members of 3 melanoma-prone families. The melanoma patients had a wide range of responsiveness to the extract, the highest responder being a 10-year survivor. As a group, family members (including spouses) without disease had significantly elevated LAI responses compared to those of unrelated controls (P less than 0.01). Within the families, members with close exposure to melanoma patients for 10 years or more had a significantly higher response to melanoma antigen than did members with 0-5 years of close exposure (P less than 0.05). Responses of spouses and members at high risk of developing melanoma (B-K mole syndrome) also correlated with length of exposure to patients, which suggests that the elevated LAI response was not genetically determined. The high frequency of positive responses to melanoma antigens in these families, particularly in spouses, suggests the presence of transmissible melanoma-associated material capable of immunizing persons in contact with melanoma patients.


The chemical reactions in electrosprays of water do not always correspond to those at the pristine air-water interface.
The recent application of electrosprays to characterize the air-water interface, along with the reports on dramatically accelerated chemical reactions in aqueous electrosprays, have sparked a broad interest. Herein, we report on complementary laboratory and in silico experiments tracking the oligomerization of isoprene, an important biogenic gas, in electrosprays and isoprene-water emulsions to differentiate the contributions of interfacial effects from those of high voltages leading to charge-separation and concentration of reactants in the electrosprays. To this end, we employed electrospray ionization mass spectrometry, proton nuclear magnetic resonance, ab initio calculations and molecular dynamics simulations. We found that the oligomerization of isoprene in aqueous electrosprays involved minimally hydrated and highly reactive hydronium ions. Those conditions, however, are non-existent at pristine air-water interfaces and oil-water emulsions under normal temperature and pressure. Thus, electrosprays should be complemented with surface-specific platforms and theoretical methods to reliably investigate chemistries at the pristine air-water interface.


Calcium release from Synechocystis cells induced by depolarization of the plasma membrane: MscL as an outward Ca2+ channel.
Cells of the cyanobacterium Synechocystis sp. PCC 6803 are equipped with a mechanosensitive ion channel MscL that is located in their plasma membrane. However, the exact function of the channel in this freshwater cyanobacterium is unknown. This study shows that cells of Synechocystis are capable of releasing Ca(2+) in response to depolarization of the plasma membrane by the K(+) ionophore valinomycin in the presence of K(+) or by tetraphenylphosphonium (TPP(+)). A fluorescent dye, diS-C(3)-(5), sensitive to membrane potential and the metallochromic Ca(2+) indicator arsenazo III were used to follow the plasma membrane depolarization and the Ca(2+) release, respectively. The Ca(2+) release from wild-type cells was temperature-dependent and it was strongly inhibited by the Ca(2+) channel blocker verapamil and by the mechanosensitive channel blocker amiloride. In MscL-deficient cells, Ca(2+) release was about 50 % of that from the wild-type cells. The mutant cells had lost temperature sensitivity of Ca(2+) release completely. However, verapamil and amiloride inhibited Ca(2+) release from these cells in same manner as in the wild-type cells. This suggests the existence of additional Ca(2+) transporters in Synechocystis, probably of a mechanosensitive nature. Evidence for the putative presence of intracellular Ca(2+) stores in the cells was obtained by following the increase in fluorescence intensity of the Ca(2+) indicator chlortetracycline. These results suggest that the MscL of Synechocystis might operate as a verapamil/amiloride-sensitive outward Ca(2+) channel that is involved in the plasma-membrane depolarization-induced Ca(2+) release from the cells under temperature stress conditions.