Girding the loins? Direct evidence of the use of a medieval English parchment birthing girdle from biomolecular analysis.
In this paper, we describe palaeoproteomic evidence obtained from a stained medieval birth girdle using a previously developed dry non-invasive sampling technique. The parchment birth girdle studied (Wellcome Collection Western MS. 632) was made in England in the late fifteenth century and was thought to be used by pregnant women while giving birth. We were able to extract both human and non-human peptides from the manuscript, including evidence for the use of honey, cereals, ovicaprine milk and legumes. In addition, a large number of human peptides were detected on the birth roll, many of which are found in cervico-vaginal fluid. This suggests that the birth roll was actively used during childbirth. This study is, to our knowledge, the first to extract and analyse non-collagenous peptides from a birth girdle using this sampling method and demonstrates the potential of this type of analysis for stained manuscripts, providing direct biomolecular evidence for active use.


MOTHERS WILLINGNESS TO VACCINATE THEIR IN-SCHOOL ADOLESCENTS AGAINST HPV INFECTION IN LAGOS STATE NIGERIA.
Human Papillomavirus (HPV) vaccine uptake rates for adolescents are still low in Nigeria despite the availability of safe and effective vaccines. The knowledge and attitudes of mothers to HPV infection and vaccines play a role in adolescents HPV vaccination uptake. This study assessed mothers' willingness to vaccinate their in-school adolescents against HPV infection in Lagos State Nigeria. A cross-sectional descriptive study involving mothers of in-school adolescents was conducted using a multi-stage sampling technique. A self-administered questionnaire was used for data collection. Chi square test was used to determine association between categorical variable and the level of significant was set at 5%. The mean age of the respondents was 43.0 ± 5.8. Majority (92%) had poor knowledge of HPV infection and vaccination. However, majority (79%) of the respondents had a positive attitude to HPV vaccination. More than half (65.8%) of the respondents' were willing to vaccinate their adolescents against HPV. There were statistically significant associations between respondents' number of daughters and attitude about HPV vaccination (p=0.021), previous pap smear screening and willingness to vaccinate their children against HPV (p=0.012) and overall knowledge of HPV vaccination and willingness to vaccinate their children against HPV (p=0.041). There is a need to increase public awareness and health education of mothers on HPV infection and cervical cancer in order to increase HPV vaccine uptake for their children. Health and education sectors can collaborate to achieve this using school-based HPV education programme.


Absolute sensitivity of electron microscope radioautography.
A calibration method is described for measuring absolute radioautographic sensitivities under various experimental conditions. Sensitivities to (3)H and (35)S radiation, i.e. ratio of developed grains to radioactive decays in the specimen, were determined with Ilford L4 and Kodak NTE emulsions. The highest values obtained in monolayers of emulsion were (1/8) for (3)H and 1/21 for (35)S. The influence of various experimental parameters on sensitivity is described, and the possibilities for quantitative electron microscope radioautography are discussed.


The avian malaria Plasmodium lophurae has a small number of heterogeneous ribosomal RNA genes.
The structure and number of the ribosomal RNA (rRNA) genes of the avian malaria parasite Plasmodium lophurae has been examined using Southern blot analysis and recombinant DNA techniques. The ribosomal DNA (rDNA) of P. lophurae has been cloned into the plasmid pBR322, beginning with size-selected populations of Cla I- and Hind III-restricted parasite DNA. The structure of two clones (CL-1 and HA-2) is presented in detail. These two clones together probably represent the entire 17s and 25s coding regions of P. lophurae. Analysis of quantitative genomic Southern blots reveals that there are approximately six rRNA genes per haploid genome, and that the rRNA genes may be divided into four distinct classes by restriction analysis. Examination of the flanking regions of these genes indicates that they are not organized into easily recognizable tandem repeats.


Enrichment, phylogenetic analysis and detection of a bacterium that performs enhanced biological phosphate removal in activated sludge.
Activated sludge communities which performed enhanced biological phosphate removal (EBPR) were phylogenetically analyzed by 16S rRNA-targeted molecular methods. Two anaerobic-aerobic sequencing batch reactors were operated with two different carbon sources (acetate vs. a complex mixture) for three years and showed anaerobic-aerobic cycles of polyhydroxybutyrate- (PHB) and phosphate-accumulation characteristic for EBPR-systems. In situ hybridization showed that the reactor fed with the acetate medium was dominated by bacteria phylogenetically related to the Rhodocyclus-group within the beta-Proteobacteria (81% of DAPI-stained cells). The reactor with the complex medium was also predominated by this phylogenetic group albeit at a lesser extent (23% of DAPI-stained cells). More detailed taxonomic information on the dominant bacteria in the acetate-reactor was obtained by constructing clone libraries of 16S rDNA fragments. Two different types of Rhodocyclus-like clones (R1 and R6) were retrieved. Type-specific in situ hybridization and direct rRNA-sequencing revealed that R6 was the type of the dominant bacteria. Staining of intracellular polyphosphate- and PHB-granules confirmed that the R6-type bacterium accumulates PHB and polyphosphate just as predicted by the metabolic models for EBPR. High similarities to 16S rDNA fragments from other EBPR-sludges suggest that R6-type organisms were present and may play an important role in EBPR in general. Although the R6-type bacterium is closely related to the genus Rhodocyclus, it did not grow phototrophically. Therefore, we propose a provisional new genus and species Candidatus Accumulibacter phosphatis.


[Dental hygienists vital to sound oral health care].
The first Dutch Dental Hygiene School was opened in 1968. Educating dental hygiene professionals with preventive qualifications was a well-considered decision. From the beginning on, the Dutch Dental Hygienists'Association has aimed at optimizing dental hygienists' role in preventive oral health care. Current developments, such as the introduction of a new national health insurance system and the rearrangement of oral health care, require reflection on the position of dental hygienists. Prevention should remain the core business of the dental hygiene profession. However, preventive oral health care has lost its social priority. Therefore, a new campaign preventing development of both old and new oral health threats should be initiated. In the opinion of the Dutch Dental Hygienists' Association, dental hygienists should be committed both to public and individual oral health prevention. The association pleads for the reinforcement of preventive oral health care for children and young adults, with a special focus on high-risk groups. Likewise, oral health care for elderly people should receive more attention.


The Station Community Mental Health Centre Inc: nurturing and empowering.
Consumer-driven community mental health services play an important role in rehabilitation, recovery, and advocacy in rural and remote Australia. The origins of services often lie in the need to provide options for people with mental illness and their carers when there is a lack of on-the-ground support. This article adds to the information about the strengths and limitations of consumer-driven mental health services by presenting the findings of an evaluation of The Station Inc. in rural South Australia. This consumer-driven mental health service provides a safe and supportive environment, social connections, and activities for its members (those with a lived experience of mental illness). Using a realist evaluation approach, the evaluation identified the contextual factors and the program mechanisms that produce positive outcomes for members. The evaluation was conducted as participatory action research with The Station members, volunteers, management committee members, and staff involved in all phases of the research process. Because of the complexity of The Station's functioning a realist evaluation using qualitative data was conducted to identify how the program worked, for whom, and in what circumstances. Twenty-five in-depth interviews were conducted with participants who were randomly selected from within the groups identified above. Interviews focused on The Station's role in assisting recovery from mental illness, the limitations and strengths of the program, and relationships with the mental health system. The Station's goals, policies and procedures, and the role of stakeholders were analysed in order to identify any links among these contextual factors, program mechanisms, and program outcomes. Qualitative data were entered into descriptive categories in N6 software (QSR; <a href="http://www.qsr.international.com" target="_blank">www.qsr.international.com). Data from the stakeholder analysis were entered into Microsoft Excel. Using an iterative approach to include the three data sets, a model was developed that identified important contextual factors that linked with two groups of program mechanisms that produced positive outcomes for members. Program mechanisms are categorised by descriptive themes referred to as 'nurturing' and 'empowering'. Nurturing' is experienced as feeling of belonging and being accepted 'as one is' and 'empowerment' mechanisms engender a belief in oneself. Respondents identified features of The Station's program, policies, atmosphere, connections and networks, stakeholder relationships, and staff and volunteers that are nurturing and empowering. Five key contextual factors enable the program mechanisms to work. The Station's coordinators ensure that nurturing and empowerment processes are highlighted through careful facilitation. The governance arrangements, policies, and administrative systems at The Station are well developed but flexibly implemented so that they support the nurturing and empowerment processes. Support and legitimacy for the program is obtained from the mental health system at state and local levels. The Station obtains resources and connections to its rural community through key stakeholders and a peak organisation One Voice Network acts as an advocate. Information about the benefits and limitations of consumer-driven mental health services in rural and remote Australia is in short supply. Increasing the available information about the contribution these services make may result in services being legitimised, understood, and resourced within mental health systems thus making the services sustainable. The benefits of consumer-driven services are that they provide flexibility and adaptation, an ability to capture the energy and passion of rural communities to improve the wellbeing of community members, and they overcome the power differential that exists between professionals and 'patients' or 'clients'.


Antigen 84, an effector of pleiomorphism in Mycobacterium smegmatis.
While in most rod-shaped bacteria, morphology is based on MreB-like proteins that form an actin-like cytoskeletal scaffold for cell wall biosynthesis, the factors that determine the more flexible rod-like shape in actinobacteria such as Mycobacterium species are unknown. Here we show that a Mycobacterium smegmatis protein homologous to eubacterial DivIVA-like proteins, including M. tuberculosis antigen 84 (Ag84), localized symmetrically to centers of peptidoglycan biosynthesis at the poles and septa. Controlled gene disruption experiments indicated that the gene encoding Ag84, wag31, was essential; when overexpressed, cells became longer and wider, with Ag84 asymmetrically distributed at one pole. Many became grossly enlarged, bowling-pin-shaped cells having up to 80-fold-increased volume. In these cells, Ag84 accumulated predominantly at a bulbous pole that was apparently generated by uncontrolled cell wall expansion. In some cells, Ag84 was associated with exceptional sites of cell wall expansion (buds) that evolved into branches. M. bovis BCG Ag84 was able to form oligomers in vitro, perhaps reflecting its superstructure in vivo. These data suggested a role for Ag84 in cell division and modulating cell shape in pleiomorphic actinobacteria.


Trials of nonoperative management exceeding 3 days are associated with increased morbidity in patients undergoing surgery for uncomplicated adhesive small bowel obstruction.
Controversy exists over how long trials of nonoperative management should be pursued in patients with uncomplicated adhesive small bowel obstructions (ASBOs) before deciding to proceed with surgery. The purpose of this study was to determine the effect of incremental delays in surgery on the 30-day postoperative outcomes of patients undergoing surgery for uncomplicated ASBO. American College of Surgeons National Surgical Quality Improvement Program 2005-2011 data were used to identify patients with uncomplicated ASBO in whom a trial of nonoperative management was attempted. Multivariate logistic or linear regression model was created to determine the independent association between the length of preoperative hospitalization and 30-day postoperative outcomes after adjustment for patient- and procedure-related factors. A total of 9,297 patients were included in the study. The 30-day postoperative mortality and overall morbidity rates of the entire cohort were 4.4% and 29.6%, respectively. The median postoperative length of hospitalization was 7 days (interquartile range, 5-11 days). After risk adjustment, there was no association between preoperative length of hospitalization and 30-day postoperative mortality. In contrast, increased 30-day overall morbidity was observed in patients who received their operation after a preoperative length of hospitalization of 3 days compared with earlier in their hospitalization. Furthermore, an increased postoperative length of hospitalization was found in patients who were operated on after a preoperative length of hospitalization of 4 days. Trials of nonoperative management for uncomplicated ASBO exceeding 3 days are associated with increased morbidity and postoperative length of hospitalization. These trials should therefore generally not extend beyond this time point. Therapeutic study, level IV.


Screening for tetrahydrobiopterin deficiency among hyperphenylalaninemia patients in Southern China.
To assess the incidence of tetrahydrobiopterin (BH4) deficiency among patients with hyperphenylalaninemia (HPA) in southern Chinese and evaluate clinical outcome and gene mutations in tetrahydrobiopterin deficient patients. Urinary neopterin (N) and biopterin (B) was analyzed in 87 patients with hyperphenylalaninemia by high-performance liquid chromatography. Further combined loading tests with phenylalanine (Phe) (100 mg/kg) and tetrahydrobiopterin (BH4) (7.5 mg/kg) were performed in suspected patients with abnormal urinary pterin profiles. Gene mutation analysis was performed for patients with BH4 deficiency and their parents. BH4 deficient patients were treated with BH4 and neurotransmitter precursors after diagnosis. Blood phenylalanine levels, clinical symptoms and mental development were followed up. Eleven patients were diagnosed as having BH4 deficiency caused by 6-pyruvoyl tetrahydropterin synthase (PTPS) deficiency. The incidence of tetrahydrobiopterin (BH4) deficiency among patients with hyperphenylalaninemia (HPA) in southern Chinese was 10%. Combined loading tests with phenylalanine and oral BH4 were done in 4 of 11 patients and their phenylalanine levels were decreased to normal 4 - 6h after BH4 administration. Four different mutations (P87S, N52S, D96N and G144R) in the PTPS gene were detected in 5 families. Five PTPS-deficient patients were treated with synthetic BH4, neurotransmitter precursors (L-dopa plus carbidopa, and 5-hydroxytryptophan). They had satisfactory physical and mental development after treatment. One patient with partial PTPS deficiency had normal growth and mental development without treatment. Our results emphasize that screening for BH4 deficiency should be carried out in all patients with hyperphenylalaninemia in order to minimize the misdiagnosis. Patients with BH4 deficiency should be treated early with BH4 and a combination of neurotransmitter precursors.


Are Plasma Oxytocin and Vasopressin Levels Reflective of Amygdala Activation during the Processing of Negative Emotions? A Preliminary Study.
Plasma oxytocin (OT) and arginine vasopressin (AVP) are associated with individual differences in emotional responses and behaviors. The amygdala is considered to be an important brain region for regulating emotion-based behavior, with OT and AVP modulating activity in the amygdala during the processing of negative emotions. In particular, increased OT levels may diminish amygdala activation (anxiolytic effects) and enhanced AVP levels may augment amygdala activation (anxiogenic effects) when negative emotions are processed. A growing body of research has shown that the effects of OT and AVP are modulated by sex: the aforementioned anxiolytic effects of OT and the anxiogenic effects of AVP occur in men, but not in women. However, we have little knowledge regarding the biological mechanisms underlying OT and AVP plasma levels or their respective anxiogenic and anxiolytic effects; similarly, little is known about the causes and nature of sex differences related to these neuropeptides and their effects on emotional processing. In the current study, we focused on the neural functions associated with the biological mechanisms underlying such effects. We hypothesized that amygdala activation would correlate with trait plasma OT (anxiolytic effects) and AVP (anxiogenic effects) levels because the amygdala is thought to affect the coordinated release of these neuropeptides following affective experiences. We further hypothesized that the effects would be modulated by sex. We assessed 51 participants (male and female) using a paradigm involving negative emotion in conjunction with functional magnetic resonance imaging and measurements of plasma OT and AVP levels. We determined that increased plasma AVP levels were positively associated with amygdala activation (anxiogenic effects) in men, but not in women. These findings highlight the potential underlying neural mechanisms of plasma AVP levels in men.


High-affinity B cell receptor ligation by cognate antigen induces cytokine-independent isotype switching.
The selection of an appropriate Ig isotype is critical for an effective immune response against pathogens. Isotype regulation is sensitive to external signals, particularly cytokines secreted by Th cells. For example, IL-4 induces isotype switching to IgG1 via a STAT6-dependent signaling pathway. In this study, we show that BCR ligation also induces IgG1 switching in mouse B cells. The extent of switch induction by Ag is affinity-dependent, and high-affinity Ag binding leads to IgG1 switching levels comparable to those induced by saturating IL-4. However, the Ag-induced IgG1 switch does not require additional cytokine signals and occurs in a STAT6-independent manner. Thus, BCR ligation represents a novel pathway for direct isotype switching leading to IgG1 secretion.


Serum 25-hydroxyvitamin D3 concentration in Iranian patients with Parkinson's disease.
Vitamin D is an important factor responsible for many physiologic functions. Vitamin D deficiency is associated with increased risk of neurodegenerative disease. The level of vitamin D in Iranian patients with Parkinson's disease and its relationship with severity of symptoms and signs were evaluated in this study. Eighty-three patients with Parkinson's disease (PD) were recruited using simple non-random sampling. 25-hydroxyvitamin D [25(OH)D3] was measured by Electrochemiluminescence immunoassay (ECLIA). Serum level of calcium and phosphorus was measured to exclude other endocrine disorders. The mean 25(OH)D3 concentration was lower in the PD population than in the normal group. Lower levels of 25(OH)D3 were associated with more severe postural instability and abnormal posture. There was no significant association between levels of 25(OH)D3 and severity of other symptoms of parkinsonism. This analysis showed that serum 25(OH)D levels are lower in PD patients in comparison with normal range. In addition, there was a significant association between the presences of freezing, postural instability and abnormal postures with lower levels of 25(OH)D.


Dyslipidemia in adults with congenital heart disease: A systematic review and meta-analysis.
Several particular characteristics of patients with congenital heart disease could affect lipid levels. The objectives of this study were: a) to analyze the prevalence of dyslipidemia in congenital heart disease patients; 2) to compare lipid levels between congenital heart disease patients and a control group. This systematic review and meta-analysis was performed according to PRISMA guidelines (PROSPERO CRD42023432041). A literature search was performed to detect studies that have reported lipid levels or the prevalence of dyslipidemia in congenital heart disease patients. We performed a qualitative analysis (studies that reported dyslipidemia prevalence) and quantitative analysis (studies that compared lipid values between congenital heart disease patients and controls). In total, 29 observational studies involving 22,914 patients with congenital heart disease and 641,086 controls were eligible for this review. The reported presence of "hyperlipidemia" or "dyslipidemia" ranged from 14.3% to 69.9%. When studies analyzed lipid variables dichotomously between congenital heart disease patients and controls, the results were conflicting. The quantitative analysis showed that patients with congenital heart disease have lower levels of total cholesterol (MD: -18.9 [95% CI: -22.2 to -15.7]; I2 = 93%), LDL-C (MD: -10.7 [95% CI: -13.1 to -8.3]; I2 = 90%) and HDL-C (MD: -6.3 [95% CI: -7.7 to -4.9]; I2 = 95%) compared to controls. The qualitative analysis showed some concerns, but the quantitative analysis indicates that congenital heart disease patients showed lower levels of total cholesterol, LDL-C, and HDL-C compared to controls. New research should be developed to clarify this relevant topic.


Reduction of Metal Artifacts and Improvement in Dose Efficiency Using Photon-Counting Detector Computed Tomography and Tin Filtration.
The aim of this study was to investigate the impact on metal artifacts and dose efficiency of using a tin filter in combination with high-energy threshold (TH) images of a photon-counting detector (PCD) computed tomography (CT) system. A 3D-printed spine with pedicle screws was scanned on a PCD-CT system with and without tin filtration. Image noise and severity of artifacts were measured for low-energy threshold (TL) and TH images. In a prospective, institutional review board-approved, Health Insurance Portability and Accountability Act-compliant study, 20 patients having a clinical energy-integrating detector (EID) CT were scanned on a PCD-CT system using tin filtration. Images were reviewed by 3 radiologists to evaluate visualization of anatomic structures, diagnostic confidence, and image preference. Artifact severity and image noise were measured. Wilcoxon signed rank was used to test differences between PCD-CT TH and EID-CT images. Phantom TH images with tin filtration reduced metal artifacts and had comparable noise (32 HU) to TL images (29 HU) acquired without tin filtration. Visualization scores for the cortex, trabeculae, and implant-trabecular interface from PCD-CT TH images (4.4 ± 0.9, 4.4 ± 1.0, and 4.4 ± 1.0) were significantly higher (P < 0.0001) than EID-CT images (3.3 ± 1.3, 3.3 ± 1.2, and 3.3 ± 1.6). A strong preference was shown for PCD-CT TH images due to improved diagnostic confidence and decreased artifact severity. Noise in PCD-CT TH images (93 ± 41 HU) was significantly lower than that in EID-CT images (133 ± 92 HU, P < 0.05). Threshold high images acquired with tin filtration on PCD-CT demonstrated a substantial decrease in metal artifacts and an increase in dose efficiency compared with EID-CT.


Fmoc-phenylalanine displays antibacterial activity against Gram-positive bacteria in gel and solution phases.
In the quest for new antimicrobial materials, hydrogels of Fmoc-protected peptides and amino acids have gained momentum due to their ease of synthesis and cost effectiveness; however, their repertoire is currently limited, and the mechanistic details of their function are not well understood. Herein, we report the antibacterial activity of the hydrogel and solution phases of Fmoc-phenylalanine (Fmoc-F) against a variety of Gram-positive bacteria including methicillin-resistant Staphylococcus aureus (MRSA). Fmoc-F, a small molecule hydrogelator, reduces the bacterial load both in vitro and in the skin wound infections of mice. The antibacterial activity of Fmoc-F is predominantly due to its release from the hydrogel. Fmoc-F shows surfactant-like properties with critical micelle concentration nearly equivalent to its minimum bactericidal concentration. Similar to Fmoc-F, some Fmoc-conjugated amino acids (Fmoc-AA) have also shown antibacterial effects that are linearly correlated with their surfactant properties. At low concentrations, where Fmoc-F does not form micelles, it inhibits bacterial growth by entering the cell and reducing the glutathione levels. However, at higher concentrations, Fmoc-F triggers oxidative and osmotic stress and, alters the membrane permeabilization and integrity, which kills Gram-positive bacteria. Herein, we proposed the use of the Fmoc-F hydrogel and its solution for several biomedical applications. This study will open up new avenues to enhance the repertoire of Fmoc-AA to act as antimicrobial agents and improve their structure-activity relationship.


Role of cellular lipoteichoic acids in mediating adherence of serotype III strains of group B streptococci to human embryonic, fetal, and adult epithelial cells.
Lipoteichoic acids (LTA) of serotype III strains of group B streptococci (GBS) were shown to mediate adherence of these organisms to human embryonic (HEC), fetal (HFC), and adult buccal (HBEC) epithelial cells. The binding of GBS was temperature dependent, and maximum attachment occurred at 37 degrees C. HEC, HFC, and HBEC preincubated with purified LTA significantly inhibited attachment of GBS, whereas the group B and type III antigens had no effect. Under phosphate-limiting conditions in which cell-associated LTA could not be detected in these organisms, bacterial adherence did not take place. GBS (virulent) that were isolated from infected infants and previously shown to have significantly higher quantities of cell-associated LTA in comparison to GBS strains from asymptomatically colonized infants adhered with greater binding avidity to HEC and HFC and in greater numbers than to HBEC. It was determined that the mechanism of LTA-mediated adherence of GBS to HBEC differed from adherence to embryonic and fetal cells for both virulent and asymptomatic GBS strains bound to HBEC in a similar manner, enhanced by the lipid portion of the LTA. In contrast, the binding of GBS to HEC and HFC was mediated by hydrophobic as well as specific interactions due to the glycerolphosphate polymer of LTA. These results indicate that possible receptor sites for LTA present on cells in prenatal stages of development may differ from those of adult cells, which may result in increased susceptibility of newborn infants to group B streptococcal disease. The implications of LTA-mediated adherence of GBS and their possible role as virulence factors are discussed.


Cortical reorganization to improve dynamic balance control with error amplification feedback.
Error amplification (EA), virtually magnify task errors in visual feedback, is a potential neurocognitive approach to facilitate motor performance. With regional activities and inter-regional connectivity of electroencephalography (EEG), this study investigated underlying cortical mechanisms associated with improvement of postural balance using EA. Eighteen healthy young participants maintained postural stability on a stabilometer, guided by two visual feedbacks (error amplification (EA) vs. real error (RE)), while stabilometer plate movement and scalp EEG were recorded. Plate dynamics, including root mean square (RMS), sample entropy (SampEn), and mean frequency (MF) were used to characterize behavioral strategies. Regional cortical activity and inter-regional connectivity of EEG sub-bands were characterized to infer neural control with relative power and phase-lag index (PLI), respectively. In contrast to RE, EA magnified the errors in the visual feedback to twice its size during stabilometer stance. The results showed that EA led to smaller RMS of postural fluctuations with greater SampEn and MF than RE did. Compared with RE, EA altered cortical organizations with greater regional powers in the mid-frontal cluster (theta, 4-7 Hz), occipital cluster (alpha, 8-12 Hz), and left temporal cluster (beta, 13-35 Hz). In terms of the phase-lag index of EEG between electrode pairs, EA significantly reduced long-range prefrontal-parietal and prefrontal-occipital connectivity of the alpha/beta bands, and the right tempo-parietal connectivity of the theta/alpha bands. Alternatively, EA augmented the fronto-centro-parietal connectivity of the theta/alpha bands, along with the right temporo-frontal and temporo-parietal connectivity of the beta band. EA alters postural strategies to improve stance stability on a stabilometer with visual feedback, attributable to enhanced error processing and attentional release for target localization. This study provides supporting neural correlates for the use of virtual reality with EA during balance training.


Detection of spatial and temporal influences on bacterial communities in an urban stream by automated ribosomal intergenic ribosomal spacer analysis.
The Yarqon is the largest urban river in Israel, and is a slow-flowing stream whose water originates mostly from wastewater treatment plants. Thus, its microbial community is expected to be heavily impacted both by anthropogenic factors and by seasonal temporal variation. In order to identify the main factors that influence the bacterial community, and their spatial-temporal variation, 50 samples were collected representing five different time points and eleven locations. Samples were analyzed for biotic and a-biotic parameters and the bacterial populations were analyzed by Automated Ribosomal Intergenic Spacer Analysis (ARISA). Bacterial richness and diversity were calculated and compared across samples. Canonical Correspondence Analysis (CCA) showed that ARISA clustered the samples according to temporal variation. Molecular fingerprinting analysis provided a snapshot of the microbial community and showed good correlation with geochemical parameters, despite the rapid changes of the Mediterranean environment and the anthropogenic impact. Molecular fingerprinting methods based on natural fragment length polymorphisms may therefore represent a supplementary approach for stream monitoring, alongside physico-chemical measurements.


Salutogenic resources in relation to teachers' work-life balance.
Experiencing work-life balance is considered a health promoting resource. To counter-balance the negative development of teachers' work situation, salutogenic resources need to be examined among teachers. To examine resources related to teachers' experience of their work-life balance. Using a cross-sectional design, a questionnaire was distributed to 455 teachers in compulsory schools in a Swedish community. A total of 338 teachers participated (74%). A multiple linear regression method was used for the analysis. Four variables in the regression model significantly explained work-life balance and were thereby possible resources: time experience at work; satisfaction with everyday life; self-rated health; and recovery. The strongest association with work-life balance was time experience at work. Except time experience at work, all were individual-related. This study highlights the importance of school management's support in reducing teachers' time pressure. It also emphasizes the need to address teachers' individual resources in relation to work-life balance. In order to support teachers' work-life balance, promote their well-being, and preventing teachers' attrition, we suggest that the school management would benefit from creating a work environment with strengthened resources.


The effects of a forgiveness intervention on patients with coronary artery disease.
This research assesses the effects of a psychology of forgiveness pilot study on anger-recall stress induced changes in myocardial perfusion, forgiveness and related variables. Thirty-two patients were administered baseline rest and anger-recall stress imaging studies, and 17 of these participants who demonstrated anger-recall stress induced myocardial perfusion defects (forgiveness group, n = 9; control group, n = 8) were randomly assigned to a series of 10 weekly interpersonal forgiveness or control therapy sessions with a trained psychologist, and underwent additional anger-recall stress myocardial perfusion nuclear imaging studies post-test and at 10-week follow-up. Patients assigned to the forgiveness group showed significantly fewer anger-recall induced myocardial perfusion defects from pre-test to the 10-week follow-up as well as significantly greater gains in forgiveness from pre-test to post-test and from pre-test to follow-up compared to the control group. Forgiveness intervention may be an effective means of reducing anger-induced myocardial ischemia in patients with coronary artery disease.


Can a comparison of clinical and deep space irradiation scenarios shed light on the radiation response of the brain?
Not surprisingly, our knowledge of the impact of radiation on the brain has evolved considerably. Decades of work have struggled with identifying the critical cellular targets in the brain, the latency of functional change and understanding how irradiation alters the balance between excitatory and inhibitory circuits. Radiation-induced cell kill following clinical fractionation paradigms pointed to both stromal and parenchymal targets but also defined an exquisite sensitivity of neurogenic populations of newly born cells in the brain. It became more and more apparent too, that acute (days) events transpiring after exposure were poorly prognostic of the late (months-years) waves of radiation injury believed to underlie neurocognitive deficits. Much of these gaps in knowledge persisted as NASA became interested in how exposure to much different radiation types, doses and dose rates that characterize the space radiation environment might impair central nervous system functionality, with possibly negative implications for deep space travel. Now emerging evidence from researchers engaged in clinical, translational and environmental radiation sciences have begun to fill these gaps and have uncovered some surprising similarities in the response of the brain to seemingly disparate exposure scenarios. This article highlights many of the commonalities between the vastly different irradiation paradigms that distinguish clinical treatments from occupational exposures in deep space.


Predictors of locoregional outcome in HER2-overexpressing breast cancer treated with neoadjuvant chemotherapy.
We identified prognostic factors for locoregional recurrence (LRR) in a cohort of patients with HER2-overexpressing breast cancer treated with neoadjuvant chemotherapy (NACT). We reviewed records of 157 patients with HER-overexpressing tumors who received NACT between May 1999 and December 2009 and collected demographics, disease/treatment characteristics, and clinical outcome. We estimated rate of LRR by the method of cumulative incidence. Presentation was 33% stage II and 67% stage III; 29.9% were clinically node positive. All patients received NACT, 94% trastuzumab containing. 90.4% had mastectomy and 6.4% breast-conserving surgery; 3.2% had no surgery. Among surgical patients, 48% were pathologically N0, 28.8% had 1 to 3 positive nodes, and 23.7% had ≥4 positive nodes. 79.6% received radiation therapy (RT) to the breast/chest wall±supraclavicular field. Median follow-up was 43 months. Three-year cumulative incidence of LRR was 8.2%; 50% of LRR had a regional component. Predictors for LRR included use of RT (HR=4.70, P=0.006), lymph node positivity (≥4 vs. 0 HR=19.99, P=0.008; 1 to 3 vs. 0 HR=10.8, P=0.031), and ER status (negative vs. positive HR=6.02, P=0.006). The only risk factor for regional failure specifically was residual nodal disease (≥4 HR=6.5, 1 to 3 HR=5.1, P=0.031). In a cohort with stage II to III HER2-overexpressing breast cancer treated predominantly with trastuzumab-containing NACT followed by mastectomy±RT, we identified omission of RT, negative ER status, and residual positive lymph nodes as significant predictors of LRR, with 50% of LRR having a regional component.


Single minimum incision endoscopic radical nephrectomy for renal tumors with preoperative virtual navigation using 3D-CT volume-rendering.
Single minimum incision endoscopic surgery (MIES) involves the use of a flexible high-definition laparoscope to facilitate open surgery. We reviewed our method of radical nephrectomy for renal tumors, which is single MIES combined with preoperative virtual surgery employing three-dimensional CT images reconstructed by the volume rendering method (3D-CT images) in order to safely and appropriately approach the renal hilar vessels. We also assessed the usefulness of 3D-CT images. Radical nephrectomy was done by single MIES via the translumbar approach in 80 consecutive patients. We performed the initial 20 MIES nephrectomies without preoperative 3D-CT images and the subsequent 60 MIES nephrectomies with preoperative 3D-CT images for evaluation of the renal hilar vessels and the relation of each tumor to the surrounding structures. On the basis of the 3D information, preoperative virtual surgery was performed with a computer. Single MIES nephrectomy was successful in all patients. In the 60 patients who underwent 3D-CT, the number of renal arteries and veins corresponded exactly with the preoperative 3D-CT data (100% sensitivity and 100% specificity). These 60 nephrectomies were completed with a shorter operating time and smaller blood loss than the initial 20 nephrectomies. Single MIES radical nephrectomy combined with 3D-CT and virtual surgery achieved a shorter operating time and less blood loss, possibly due to safer and easier handling of the renal hilar vessels.


Mothers' beliefs about analgesia during childhood immunization.
Immunization injections are the most common painful medical procedures experienced during childhood, yet there is a discrepancy between recommendations for the effective use of topical anesthetics to reduce vaccine injection pain and actual practice. To improve our understanding of mothers' experiences and practices regarding their children's routine immunizations. Adopting an interpretive, naturalistic paradigm, semi-structured interviews were conducted with 15 mothers to examine their perceptions and experiences of their children's immunization pain and pain management. The findings demonstrated three main themes: attitudes toward immunization pain, immunization pain management and physicians as sources of information. Participants described feeling distressed while their children were being immunized, but most managed these difficulties by focusing on the benefits of immunization and by minimizing or justifying the pain. All of the participants used non-pharmacological techniques to manage immunization injection pain. Few mothers were aware of the availability of topical anesthetics. When participants did use pharmacological analgesic approaches, oral analgesics were most likely to be used for prophylaxis and treatment of fever, and participants were unaware of evidence-based approaches to managing pain. Participants viewed their physicians as trusted sources of information, and the majority said that they would likely use a topical anesthetic in the future if recommended or approved by their physician. The present findings provide direction for future knowledge translation activities to enhance the knowledge of mothers and clinicians regarding pain during immunization injections and its effective management.


Secondary infection presenting as recurrent pulmonary hypertension.
Primary infection in the neonate, especially group B streptococcal infection, has long been recognized as a cause of persistent pulmonary hypertension of the newborn (PPHN), sometimes requiring treatment with inhaled nitric oxide (iNO) and extracorporeal membrane oxygenation (ECMO). However, secondary nosocomial infections in the neonatal period have not been widely reported as a cause of severe recurrent pulmonary hypertension (PHTN). We now present two cases of secondary infection in the neonate leading to significant PHTN. In both cases, the infants presented with PPHN soon after birth, requiring transfer to a level 3 neonatal intensive care unit and treatment with high-frequency oscillatory ventilation and iNO. After successful resolution of the initial PPHN, including extubation to nasal cannula, both infants developed signs of severe recurrent PHTN, leading to reintubation, high-frequency oscillatory ventilation and iNO therapy, and consideration of ECMO. In both cases, blood cultures taken at the time of recurrence of PHTN returned positive, one for Staphylococcus epidermidis, the other for methicillin-resistant Staphylococcus aureus. These unusual cases present the possibility of severe recurrent PHTN requiring iNO or ECMO in the setting of secondary infection. We speculate that these infants, although extubated after their first episodes of PHTN, were at risk for recurrence of PHTN due to continued pulmonary vascular reactivity.


Variation in Protein Origin and Utilization: Research and Clinical Application.
Muscle health can be rapidly compromised in clinical environments. Modifiable strategies to preserve metabolic homeostasis in adult patient populations include physical activity and pharmacologic support; however, optimizing dietary practices, or more specifically protein intake, is a necessary prerequisite for any other treatment strategy to be fully effective. Simply increasing protein intake is a well-intentioned but often unfocused strategy to protect muscle health in an intensive care setting. Protein quality is a frequently overlooked factor with the potential to differentially influence health outcomes. Quality can be assessed by a variety of techniques, with digestible indispensable amino acid score being the current and most comprehensive technique endorsed by the Food and Agriculture Organization. In practical terms, animal-based proteins are consistently scored higher in quality compared with incomplete proteins, regardless of the assessment method. Consequently, choosing parenteral and/or enteral feeding options that contain high-quality proteins, rich in the branched-chain amino acid leucine, may help establish a dietary framework with the potential to support clinical practice and improve health outcomes in critically ill patients.


Relation of animal size to convergence, divergence, and neuronal number in peripheral sympathetic pathways.
The enormous range of animal size raises a fundamental problem: How do larger animals maintain adequate control of peripheral structures that are many times more massive and extensive than the homologous structures in smaller animals? To explore this question, we have determined neuronal number, the number of axons that innervate each neuron (convergence) and the number of neurons innervated by each axon (divergence), in a peripheral sympathetic pathway of several mammals (mouse, hamster, rat, guinea pig, and rabbit). The average adult weights of these species vary over approximately a 65-fold range. However, the number of superior cervical ganglion cells increases by only a factor of 4 between the smallest of these animals (mice; about 25 gm) and the largest (rabbits; about 1700 gm); the number of spinal preganglionic neurons that innervate the ganglion increases by only a factor of 2. Thus, the number of nerve cells in the sympathetic system does not increase in proportion to animal size. On the other hand, our results indicate that there are systematic differences across these species in the number of axons that innervate each ganglion cell and in the number of ganglion cells innervated by each axon. We suggest that modulation of convergence and divergence in sympathetic ganglia allows this part of the nervous system to effectively activate homologous peripheral targets over a wide range of animal size.


Children Persist Less When Adults Take Over.
Children need to learn to persist through challenges, yet adults sometimes step in to solve problems for them. Here, we looked at how adult taking over related to children's persistence. In an observational study (N = 34, ages 4-8), we found that parents who took over more often during a challenging puzzle task rated their children as dispositionally less persistent. To establish whether taking over can cause reduced persistence, we ran two preregistered experiments (N = 150, ages 4-5). Children assigned to a taking over condition persisted less on a subsequent task compared to those in a teaching or a baseline condition. Reframing the context did not ameliorate the negative impact of taking over. The results suggest that taking over impairs children's persistence.


Hysteretic DC electrowetting by field-induced nano-structurations on polystyrene films.
Electrowetting (EW) offers executive wetting control of conductive liquids on several polymer surfaces. We report a peculiar electrowetting response for aqueous drops on a polystyrene (PS) dielectric surface in the presence of silicone oil. After the first direct current (DC) voltage cycle, the droplet failed to regain Young's angle, yielding contact angle hysteresis, which is close to a value found in ambient air. We conjecture that the hysteretic EW response appears from in situ surface modification using electric field induced water-ion contact with PS surface inducing nano-structuration by electro-hydrodynamic (EHD) instability. Atomic force microscopy confirms the formation of nano-structuration on the electrowetted surface. The effects of molecular weight, applied electric field, water conductivity and pH on nano-structuration are studied. Finally, the EW based nano-structuration on PS surface is used for the enhanced loading of aqueous dyes on hydrophobic surfaces.


Do sympathetic neurons coordinate cellular development in the heart and kidney? Effects of neonatal central and peripheral catecholaminergic lesions on cardiac and renal nucleic acids and proteins.
Sympathetic nerve activity has been hypothesized to set the timing of cellular maturational events in target tissues. In the current study, this hypothesis was tested by lesioning catecholamine pathways in the periphery and central nervous system through the use of subcutaneous or intracisternal injections of 6-hydroxydopamine. Systemically administered 6-hydroxydopamine completely depleted peripheral norepinephrine. The central treatment completely ablated the developmental rise in brain norepinephrine and dopamine and had little effect on peripheral norepinephrine levels, but has been shown to reduce sympathetic tone. In both the heart and the kidney, either type of lesion resulted in deficits in cell acquisition (DNA) with some evidence of compensatory increases in other macromolecules involved in cell enlargement (particularly RNA), thus maintaining the tissue growth rate at only slightly subnormal levels. The peak effect was always seen during the stages at which sympathetic neuronal synaptogenesis and impulse activity ordinarily undergo their most rapid development. Most of the 6-hydroxydopamine-induced differences in nucleic acids lessened or disappeared toward weaning, and thus these data support the view that sympathetic neuronal input influences the timing of maturational control of macromolecules, but not their final set-point. In combination with earlier studies showing termination of DNA synthesis caused by exposure of heart and kidney acutely to high levels of catecholamines, the results suggest that neuronal activity provides a biphasic signal, with positive trophic effects predominating during early development when sympathetic tone is low, and negative effects appearing when sympathetic tone is elevated during the late preweanling stage.


Impact of BNT162b2 mRNA anti-SARS-CoV-2 vaccine on interferon-alpha production by plasmacytoid dendritic cells and autoreactive T cells in patients with systemic lupus erythematosus: The COVALUS project.
To evaluate the specific response of SLE patients to BNT162b2 vaccination and its impact on autoimmunity defined as in vivo production of interferon-alpha (IFNα) by plasmacytoid dendritic cells (pDCs) and autoreactive immune responses. Our prospective study included SLE patients and healthy volunteers (HV) who received 2 doses of BNT162b2 vaccine 4 weeks apart. Subjects under immunosuppressive drugs or with evidence of prior COVID-19 were excluded. IgG anti-Spike SARS-CoV-2 (anti-S) antibodies, anti-S specific-B cells, anti-S specific T cells, in vivo INF-α production by pDCs, activation marker expression by pDCs and autoreactive anti-nuclear T cells were quantified before first injection, before second injection, and 3 and 6 months after first injection. Vaccinated SLE patients produced significantly lower IgG antibodies and specific B cells against SARS-CoV-2 as compared to HV. In contrast, anti-S T cell response did not significantly differ between SLE patients and HV. Following vaccination, the surface expression of HLA-DR and CD86 and the in vivo production of IFNα by pDCs significantly increased in SLE patients. The boosted expression of HLA-DR on pDCs induced by BNT162b2 vaccine correlated with the overall immune responses against SARS-CoV-2 (anti-S antibodies: r = 0.27 [0.05-0.46], p = 0.02; anti-S B cells: r = 0.19 [-0.03-0.39], p = 0.09); anti-S T cells: r = 0.28 [0.05-0.47], p = 0.016). Eventually, anti-SARS-CoV-2 vaccination was associated with an overall decrease of autoreactive T cells (slope = - 0.00067, p = 0.015). BNT162b2 vaccine induces a transient in vivo activation of pDCs in SLE that contributes to the immune responses against SARS-CoV-2. Unexpectedly BNT162b2 vaccine also dampens the pool of circulating autoreactive T cells, suggesting that vaccination may have a beneficial impact on SLE disease.


Relationship between the hypothalamic-pituitary-thyroid axis and alcohol craving in alcohol-dependent patients: a longitudinal study.
A relationship between some hypothalamic-pituitary-related hormones and craving for alcohol has been suggested, leading to hypothesize a role of some hormones in the neurobiology of alcohol dependence. Investigating this association in alcohol-dependent (AD) patients was the aim of this preliminary and exploratory study. Cortisol, adrenocorticotropic hormone, prolactin, thyroid-stimulating hormone (TSH), free T3, free T4, growth hormone, follicle-stimulating hormone, luteinizing hormone as well as administering the Obsessive-Compulsive Drinking Scale (OCDS) and Penn Alcohol Craving Scale (PACS) were assessed at baseline and after 12 weeks in 25 current AD patients. Patients were treated with baclofen (10 mg t.i.d.) for these 12 weeks. Sixteen patients remained totally abstinent for 12 weeks. At baseline, a significant inverse correlation was found between TSH and PACS (r = -0.46; p = 0.022) and OCDS scores (r = -0.53; p = 0.007). A significant direct correlation was found between free T3 and OCDS score (r = 0.44; p = 0.026). In the 16 abstinent patients, craving scores were significantly decreased at 12 weeks (p < 0.01). At 12 weeks, no significant correlation was found between TSH and craving, but free T3 remained directly correlated with OCDS score (r = 0.60; p = 0.013). A relationship between alcohol craving and free T3 and TSH was demonstrated in AD patients, suggesting the potential involvement of the hypothalamic-pituitary-thyroid axis in the neurobiology of alcohol craving.


Involvement of staphylococcal protein A and cytoskeletal actin in Staphylococcus aureus invasion of cultured human oral epithelial cells.
Following the coincidental discovery that beta-actin isolated from renal epithelial cells was precipitated by staphylococcal protein A (SPA), the possibility that SPA and cytoskeletal actin filaments may be involved in Staphylococcus aureus infection of epithelial cells was considered. Therefore, to clarify the potential role of SPA and actin filaments in S. aureus infection, the invasion efficiency of S. aureus was determined quantitatively by measuring the number of cfu of viable organisms recovered from cultured KB cells. S. aureus invasion was found to be time dependent (0-60 min) and increased linearly when increasing numbers of bacteria were added (10(4)-10(6) cfu/ml). However, significant variation in the level of invasion was noted in protein A-deficient S. aureus Wood 46. Cytochalasin B inhibited the invasion efficiency of S. aureus in a dose-dependent manner. The present study suggests that interaction of staphylococcal protein A and cytoskeletal actin filaments is involved in the S. aureus invasion of cultured KB cells, and this process may contribute, in part, to the intracellular movement, cell-to-cell spread and dissemination of S. aureus within human oral epithelial cells in vivo.


Conceiving wholeness: women, motherhood, and ovarian transplantation, 1902 and 2004.
Scholars have shown that organ transplantation may transform ideas about one's body, with recipients feeling that they are receiving not just a body part but also a part of the donor's identity. This article focuses on a different way in which organ transplantation shapes recipient identity: the idea of becoming whole. We present the case studies of two women separated by a century (one in 1902 and the other in 2004) who sought ovarian transplantation, and examine how ovarian transplantation can engender a sense of wholeness on the individual, the familial, and the cultural levels, due to its ability to enable a recipient to naturally conceive and experience pregnancy.


NAP (davunetide) provides functional and structural neuroprotection.
NAP (davunetide) is an eight amino acid peptide (NAPVSIPQ) that has been shown to provide potent neuroprotection, in vitro and in vivo. In human clinical trials, NAP has been shown to increase memory scores in patients suffering from amnestic mild cognitive impairment, a precursor to Alzheimer's disease and to enhance functional daily behaviors in schizophrenia patients. NAP is derived from activity-dependent neuroprotective protein (ADNP) a molecule that is essential for brain formation, interacting with chromatin associated protein alpha and the chromatin remodeling complex SWI/SNF and regulating >400 genes during embryonic development. Partial loss in ADNP results in cognitive deficits and pathology of the microtubule associated protein tau (tauopathy) that is ameliorated in part by NAP replacement therapy. Recent studies increased the scope of NAP neuroprotection and provided further insights into the NAP mechanisms of action. Thus, it has been hypothesized that the presence of tau on axonal microtubules renders them notably less sensitive to the microtubule-severing protein katanin, and NAP was shown to protect microtubules from katanin disruption in the face of reduced tau expression. Parallel studies showed that NAP reduced the number of apoptotic neurons through activation of PI-3K/Akt pathway in the cortical plate or both PI-3K/Akt and MAPK/MEK1 kinases in the white matter. The interaction of these disparate yet complementary pathways is the subject of future studies toward human brain neuroprotection in the clinical scenario.


Handwashing and risk of respiratory infections: a quantitative systematic review.
To determine the effect of handwashing on the risk of respiratory infection. We searched PubMed, CAB Abstracts, Embase, Web of Science, and the Cochrane library for articles published before June 2004 in all languages. We had searched reference lists of all primary and review articles. Studies were included in the review if they reported the impact of an intervention to promote hand cleansing on respiratory infections. Studies relating to hospital-acquired infections, long-term care facilities, immuno-compromised and elderly people were excluded. We independently evaluated all studies, and inclusion decisions were reached by consensus. From a primary list of 410 articles, eight interventional studies met the eligibility criteria. All eight eligible studies reported that handwashing lowered risks of respiratory infection, with risk reductions ranging from 6% to 44% [pooled value 24% (95% CI 6-40%)]. Pooling the results of only the seven homogenous studies gave a relative risk of 1.19 (95% CI 1.12%-1.26%), implying that hand cleansing can cut the risk of respiratory infection by 16% (95% CI 11-21%). Handwashing is associated with lowered respiratory infection. However, studies were of poor quality, none related to developing countries, and only one to severe disease. Rigorous trials of the impact of handwashing on acute respiratory tract infection morbidity and mortality are urgently needed, especially in developing countries.


[New potential of estrogen and progesterone receptor assay in cytological smears sampled from breast cancer patients].
Estrogen (ER) and progesterone (PR) receptors were assayed immunohistochemically in cytological smears and histological samples from 30 breast cancer patients. Since the results reported by the two research centers matched by 73%, it is suggested that running ER and PR determinations on cytological smears in cases of inoperable cancer, preoperative therapy and monitoring may be recommended. Therefore, the 3-4 year old records on ER and PR determinations should be regarded as null and void.


HIV prevention for people with serious mental illness: a survey of mental health workers' attitudes, knowledge and practice.
The aim of this survey was to investigate the attitudes, knowledge and reported practice (capabilities) of mental health workers concerning human immunodeficiency virus (HIV) and other sexually transmitted diseases in people with serious mental illness. People with serious mental illness are at increased risk of HIV and other sexually transmitted infections. Mental health workers have a key role to play in promoting sexual health in this population, but it is unclear how they perceive their role in this work and whether they have the capabilities to deliver sexual health promotion. Cross sectional survey. A questionnaire was devised and distributed to 650 mental health workers working in a London (UK) NHS mental health service. A response rate of 44% was achieved. Overall, workers reported positive attitudes to sexual health promotion and were knowledgeable about risk behaviours and risk factors for HIV infection. Adherence to glove wearing was good. However, participants' knowledge about HIV/AIDS in people with schizophrenia was poor and most reported they were not engaged in sexual health promotion activities with people with serious mental illness. Glove wearing was predicted by those who had drug and alcohol training and clinical experience and knowledge of risk factors was predicted by previous health promotion training. No other demographic factors predicted any of the other subscales. Mental health workers require training to provide skills for health promotion regarding sexual health and HIV in people with serious mental health problems. In addition, there needs to be more research on risk behaviours. The development of effective interventions to reduce this behaviour.


The structure of the catalytic domain of Tannerella forsythia karilysin reveals it is a bacterial xenologue of animal matrix metalloproteinases.
Metallopeptidases (MPs) are among virulence factors secreted by pathogenic bacteria at the site of infection. One such pathogen is Tannerella forsythia, a member of the microbial consortium that causes peridontitis, arguably the most prevalent infective chronic inflammatory disease known to mankind. The only reported MP secreted by T. forsythia is karilysin, a 52 kDa multidomain protein comprising a central 18 kDa catalytic domain (CD), termed Kly18, flanked by domains unrelated to any known protein. We analysed the 3D structure of Kly18 in the absence and presence of Mg(2+) or Ca(2+) , which are required for function and stability, and found that it evidences most of the structural features characteristic of the CDs of mammalian matrix metalloproteinases (MMPs). Unexpectedly, a peptide was bound to the active-site cleft of Kly18 mimicking a left-behind cleavage product, which revealed that the specificity pocket accommodates bulky hydrophobic side-chains of substrates as in mammalian MMPs. In addition, Kly18 displayed a unique Mg(2+) or Ca(2+) binding site and two flexible segments that could play a role in substrate binding. Phylogenetic and sequence similarity studies revealed that Kly18 is evolutionarily much closer to winged-insect and mammalian MMPs than to potential bacterial counterparts found by genomic sequencing projects. Therefore, we conclude that this first structurally characterized non-mammalian MMP is a xenologue co-opted through horizontal gene transfer during the intimate coexistence between T. forsythia and humans or other animals, in a very rare case of gene shuffling from eukaryotes to prokaryotes. Subsequently, this protein would have evolved in a bacterial environment to give rise to full-length karilysin that is furnished with unique flanking domains that do not conform to the general multidomain architecture of animal MMPs.


Catalytic Mechanism of Aromatic Nitration by Cytochrome P450 TxtE: Involvement of a Ferric-Peroxynitrite Intermediate.
The cytochromes P450 are heme-dependent enzymes that catalyze many vital reaction processes in the human body related to biodegradation and biosynthesis. They typically act as mono-oxygenases; however, the recently discovered P450 subfamily TxtE utilizes O2 and NO to nitrate aromatic substrates such as L-tryptophan. A direct and selective aromatic nitration reaction may be useful in biotechnology for the synthesis of drugs or small molecules. Details of the catalytic mechanism are unknown, and it has been suggested that the reaction should proceed through either an iron(III)-superoxo or an iron(II)-nitrosyl intermediate. To resolve this controversy, we used stopped-flow kinetics to provide evidence for a catalytic cycle where dioxygen binds prior to NO to generate an active iron(III)-peroxynitrite species that is able to nitrate l-Trp efficiently. We show that the rate of binding of O2 is faster than that of NO and also leads to l-Trp nitration, while little evidence of product formation is observed from the iron(II)-nitrosyl complex. To support the experimental studies, we performed density functional theory studies on large active site cluster models. The studies suggest a mechanism involving an iron(III)-peroxynitrite that splits homolytically to form an iron(IV)-oxo heme (Compound II) and a free NO2 radical via a small free energy of activation. The latter activates the substrate on the aromatic ring, while compound II picks up the ipso-hydrogen to form the product. The calculations give small reaction barriers for most steps in the catalytic cycle and, therefore, predict fast product formation from the iron(III)-peroxynitrite complex. These findings provide the first detailed insight into the mechanism of nitration by a member of the TxtE subfamily and highlight how the enzyme facilitates this novel reaction chemistry.


Regional differences in stroke mortality and alcohol consumption in Japan.
The relationship between alcohol consumption and stroke mortality in 1975 in 46 prefectures of Japan was investigated. This was done by adjusting salt intake and several socio-economic factors, i.e., the annual per capita income, the number of persons who received public aid, the number of tatamis (a Japanese traditional floor unit) per household, the unemployment rate, and the unmarried or divorce rate, using a stepwise multiple regression analysis. As dependent variables, the sex-specific and age-adjusted mortality for the middle-aged (35-59 years) and for all ages due to stroke were used. For men, alcohol consumption was significantly related to age-adjusted stroke mortalities for the middle-aged and for all ages independent of salt intake and several socio-economic factors. Alcohol consumption was more strongly related to age-adjusted stroke mortality for the middle-aged than for all ages. For women alcohol was weakly correlated with the stroke mortality of the middle-aged. Salt intake was significantly correlated with stroke mortality for women but not for men. Furthermore, the male: female ratios of the age-adjusted stroke mortality for the middle aged and for all ages were analyzed as well, because alcohol is mostly consumed by men in Japan, and it was expected that the sex ratios would be well correlated to alcohol consumption. The results were as expected. Therefore, it was suggested that the regional difference in stroke mortality in Japan may be explained in part by that of alcohol consumption.


Vulnerability to cardiac disease.
Chronic consumption of ethanol has deleterious effects on the cardiovascular system, as manifested by an attenuation in myocardial contractility, a reduction in cardiac output, and the induction of arrhythmia. The arrhythmogenic effect of ethanol is associated with the high incidence of sudden death in alcoholics. Further, alcohol was found to potentiate arrhythmias due to nonpenetrating chest trauma, a finding of profound clinical significance. In addition, chronic ethanol consumption is closely linked to hypertension. Whether modest alcohol consumption may protect against coronary artery disease is controversial and not clearly established. Cessation of alcohol consumption occasionally results in reversal of ethanol-induced myocardial injury. However, the transition from ethanol-induced reversible injury to permanent heart damage is not well understood. Finally, the combined effects on the myocardium of alcohol and other abused drugs, such as cocaine and amphetamines, and the interaction of ethanol with chemicals such as nicotine, digitalis, and other medicaments are not well understood and may be fatal.


Differentiation of human fibroblasts to tissue macrophages by the Snyder-Theilen feline sarcoma virus (ST:FeSV).
We have previously reported the conversion of human fibroblasts (HF) to tissue macrophages (TM) by transduction with the Snyder-Theilen feline sarcoma virus [ST:FeSV(FeLV)]. Here, we have demonstrated macrophage mediated tumor cytotoxicity (MTC) of human teratocarcinoma cells (NTERA-2) by ST:FeSV-induced TM. Inclusion of the NTERA-2 specific antibody (SSEA-4), substantially increased antibody-dependent cellular cytotoxicity (ADCC) of the tumor cells by ST:FeSV-induced TM. Tumor cell lysis by ST:FeSV-induced TM through either MTC or ADCC was especially effective at low serum concentrations (1%). The presence of cytokines during co-incubation, M-CSF in particular, enhanced the lysis of NTERA-2 cells by ST:FeSV-induced TM. Whereas pretreatment of the NTEBA-2 cells with all-trans retinoic acid (AtRA) enhanced MTC-induced lysis, it abrogated ADCC-induced lysis of the tumor cells by ST:FeSV-induced TM. Significantly, the co-incubation of NTERA-2 cells with ST:FeSV-induced TM in agar, resulted in a considerable stimulation of growth of untreated and TPA-pretreated NTERA-2 cells. In contrast, the growth of AtRA-pretreated NTERA-2 cells in agar was completely inhibited by ST:FeSV-induced TM. These results indicate that ST:FeSV-induced TM can produce potent growth-modulating cytokines, and that AtRA can modify their antitumor activity in monolayer cultures and in agar. The effects by AtRA, serum, and growth conditions (whether monolayers or agar) point to several levels of complexity regarding the interaction of tissue macrophages with tumor cells.


Point Prevalence and Patterns of Mental Health Comorbidity Among People Accessing Australia's First Older Adult-Specific Alcohol and Other Drug Treatment Service.
There are good data regarding the prevalence and patterns of dual diagnosis among the general population; however, data regarding the older adult cohort are limited. We aimed to extend the knowledge of the point prevalence and patterns of dual diagnosis among older adults and the impact of dual diagnosis on the utilization of alcohol and other drug treatment services. A 12-month medical chart audit of clients discharged from an Australian older adult-specific alcohol and other drug treatment service was performed. Measures included the Alcohol Use Disorders Identification Test-Consumption, the Drug Use Disorders Identification Test-Consumption, the Kessler 10, and the Modified MINI Screen. Additional data collected included mental health diagnoses, number of session types, and treatment outcomes. There were 79 (n = 45, 57% male) medical charts audited, with a mean age of 65.9 years (SD = 5.8). There were 68 (89%) clients having at least one comorbid mental illness. Clients with a dual diagnosis were younger (p = .011) than those without. Some comorbid mental health conditions were associated with additional service utilization (p < .05). Clients with personality disorders required more telephone calls and outreach services (p < .05). The number of mental health diagnoses was associated with additional treatment sessions (p < .05). Further research with a larger sample size of older adults seeking age-specific alcohol and other drug treatment services is required. Older adult-specific alcohol and other drug treatment services need to allow for longer episodes of care for clients with certain dual diagnoses and a focus on reducing anxiety to increase treatment retention.


TDP-43 proteinopathy impairs mRNP granule mediated postsynaptic translation and mRNA metabolism.
Background: Local protein synthesis and mRNA metabolism mediated by mRNP granules in the dendrites and the postsynaptic compartment is essential for synaptic remodeling and plasticity in neuronal cells. Dysregulation of these processes caused by TDP-43 proteinopathy leads to neurodegenerative diseases, such as frontotemporal lobar degeneration and amyotrophic lateral sclerosis. Methods: Using biochemical analysis and imaging techniques, including super-resolution microscopy, we provide evidence, for the first time, for the postsynaptic localization of TDP-43 in mammalian synapses and we show that TDP-43 is a component of neuronal mRNP granules. Results: With activity stimulation and various molecular approaches, we further demonstrate activity-dependent mRNP granule dynamics involving disassembly of mRNP granules, release of mRNAs, activation of local protein translation, and the impairment of granule disassembly in cellular, animal and human models of TDP-43 proteinopathy. Conclusion: Our study elucidates the interplay between TDP-43 and neuronal mRNP granules in normal physiology and TDP-43 proteinopathy in the regulation of local protein translation and mRNA metabolism in the postsynaptic compartment.


[The composition of lipids and lipid peroxidation in the pancreas of quails exposed to nitrates and correction by the amaranth's seeds].
Researches of features of lipid composition, functioning of the system of antioxidant defense, maintenance of lipid peroxidation products in the quail's pancreas on the early postnatal ontogenesis stages are conducted for actions of nitrates and feeding with amaranth's seeds in mixed fodder. The arrival of nitrates in the organism of quails results in the decline of general lipids maintenance and nonetherified fat acids in the pancreas. Using of amaranth's seeds in mixed fodder on the background of the nitrate loading results in the increase of activity of the enzimes system of antioxidant defence, the growth of general lipid level in the quail's pancreas. Thus in correlation with separate classes of lipid maintenance of cholesterol goes down for certain, whereas the maintenance of triacylglycerols and ethers of cholesterol rises. The results obtained in the researches show the ability of amaranth's seeds to avert oxidative stress in quail's pancreas under nitrates influence.


Mechanisms Mediating Functional Hyperemia in the Brain.
Neuronal activity within the brain evokes local increases in blood flow, a response termed functional hyperemia. This response ensures that active neurons receive sufficient oxygen and nutrients to maintain tissue function and health. In this review, we discuss the functions of functional hyperemia, the types of vessels that generate the response, and the signaling mechanisms that mediate neurovascular coupling, the communication between neurons and blood vessels. Neurovascular coupling signaling is mediated primarily by the vasoactive metabolites of arachidonic acid (AA), by nitric oxide, and by K+. While much is known about these pathways, many contentious issues remain. We highlight two controversies, the role of glial cell Ca2+ signaling in mediating neurovascular coupling and the importance of capillaries in generating functional hyperemia. We propose signaling pathways that resolve these controversies. In this scheme, capillary dilations are generated by Ca2+ increases in astrocyte endfeet, leading to production of AA metabolites. In contrast, arteriole dilations are generated by Ca2+ increases in neurons, resulting in production of nitric oxide and AA metabolites. Arachidonic acid from neurons also diffuses into astrocyte endfeet where it is converted into additional vasoactive metabolites. While this scheme resolves several discrepancies in the field, many unresolved challenges remain and are discussed in the final section of the review.


Increased and sex-selective avian predation of desert locusts Schistocerca gregaria treated with Metarhizium acridum.
The entomopathogenic fungus Metarhizium acridum in oil-based formulations (Green Muscle® (GM)) is a biopesticide for locust control lacking side-effects on biodiversity, unlike chemical insecticides. Under controlled conditions, GM-treated locusts and grasshoppers attract predators, a complementary advantage in locust control. We assessed avian predation on a population of desert locusts in northern Niger aerially sprayed operationally with GM with 107 g viable conidia ha-1. Populations of adult locusts and birds and vegetation greenness were assessed simultaneously along two transects from 12 days before until 23 days after treatment. Common kestrels Falco tinnunculus and lanners F. biarmicus were the predominant avian predators. Regurgitated pellets and prey remains were collected daily beneath "plucking posts" of kestrels. Locusts started dying five days post-spray and GM had its maximum effect one-two weeks after the spray, with 80% efficacy at day 21. After spraying, bird numbers increased significantly (P<0.05) concurrent with decreasing desert locust densities. Locust numbers decreased significantly (P<0.001) with both time since spraying and decreasing greenness. Before spraying, kestrel food remains under plucking posts accounted for 34.3 ±13.4 prey items day-1, of which 31.0 ±11.9 were adult desert locusts (90.3%), reducing post-spray to 21.1 ±7.3 prey items day-1, of which19.5 ±6.7 were adult desert locusts (92.5%), attributable to decreased use of the plucking-posts by the kestrels rather than an effect of the spray. After spraying, kestrels took significantly (P<0.05) more larger female (75-80%) than smaller male (20-25%) locusts. Avian predation probably enhanced the impact of the GM on the desert locust population, especially by removing large adult females. No direct or indirect adverse side-effects were observed on non-target organisms including locust predators such as ants and birds. These substantial ecological advantages should also be considered when choosing between conventional chemical and biopesticide-based locust control.


Interactions of murine APOBEC3 and human APOBEC3G with murine leukemia viruses.
APOBEC3 proteins are cytidine deaminases which help defend cells against retroviral infections. One antiviral mechanism involves deaminating dC residues in minus-strand DNA during reverse transcription, resulting in G-to-A mutations in the coding strand. We investigated the effects of mouse APOBEC3 (mA3) and human APOBEC3G (hA3G) upon Moloney murine leukemia virus (MLV). We find that mA3 inactivates MLV but is significantly less effective against MLV than is hA3G. In contrast, mA3 is as potent against human immunodeficiency virus type 1 (HIV-1, lacking the protective Vif protein) as is hA3G. The two APOBEC3 proteins are packaged to similar extents in MLV particles. Dose-response profiles imply that a single APOBEC3 molecule (or oligomer) is sufficient to inactivate an MLV particle. The inactivation of MLV by mA3 and hA3G is accompanied by relatively small reductions in the amount of viral DNA in infected cells. Although hA3G induces significant levels of G-to-A mutations in both MLV and HIV DNAs, and mA3 induces these mutations in HIV DNA, no such mutations were detected in DNA synthesized by MLV inactivated by mA3. Thus, MLV has apparently evolved to partially resist the antiviral effects of mA3 and to totally resist the ability of mA3 to induce G-to-A mutation in viral DNA. Unlike the resistance of HIV-1 and human T-cell leukemia virus type 1 to hA3G, the resistance of MLV to mA3 is not mediated by the exclusion of APOBEC from the virus particle. The nature of its resistance and the mechanism of inactivation of MLV by mA3 are completely unknown.


Preferential prey selection by Desmodus rotundus (E. Geoffroy, 1810, Chiroptera, Phyllostomidae) feeding on domestic herbivores in the municipality of São Pedro--SP.
In order to verify possible preferential prey selection by Desmodus rotundus feeding on domestic herbivores in the Municipality of São Pedro (São Paulo, Brazil), vampire bat attacks were surveyed at rural properties where domestic herbivores were being raised and attack frequencies of D. rotundus on the total herd and on different species were calculated. The analysis found that the most frequently attacked herbivores were cattle and horses. The chi-square test (χ2), with a significance level of 5% corroborated the comparative analysis of attack frequency in properties that had these two species. Of the two, horses were attacked more frequently, which could be a sign that D. rotundus exhibits preferential prey selection when attacking domestic herbivores. This evidence is also supported by the Optimal Foraging Theory, in which the net rate of energy consumed is higher for horses than it is for cattle. Additionally, we propose that the thinner integument of horses (relative to that of cattle) may facilitate bites by D. rotundus and thus contribute to the observed prey preference.


Transposed-letter priming of prelexical orthographic representations.
A prime generated by transposing two internal letters (e.g., jugde) produces strong priming of the original word (judge). In lexical decision, this transposed-letter (TL) priming effect is generally weak or absent for nonword targets; thus, it is unclear whether the origin of this effect is lexical or prelexical. The authors describe the Bayesian Reader theory of masked priming (D. Norris & S. Kinoshita, 2008), which explains why nonwords do not show priming in lexical decision but why they do in the cross-case same-different task. This analysis is followed by 3 experiments that show that priming in this task is not based on low-level perceptual similarity between the prime and target, or on phonology, to make the case that priming is based on prelexical orthographic representation. The authors then use this task to demonstrate equivalent TL priming effects for nonwords and words. The results are interpreted as the first reliable evidence based on the masked priming procedure that letter position is not coded absolutely within the prelexical, orthographic representation. The implications of the results for current letter position coding schemes are discussed.


The antiquity of hydrocephalus: the first full palaeo-neuropathological description.
The Pathology Museum of the University of Florence houses a rich collection of anatomical specimens and over a hundred waxworks portraying pathological conditions occurring in the nineteenth century, when the museum was established. Clinical and autopsy findings of these cases can still be retrieved from the original museum catalogue, offering a rare opportunity for retrospective palaeo-pathological diagnostics. We present a historical case of severe hydrocephalus backed by modern-day anthropological, radiological and molecular analyses conducted on the skeleton of an 18-month-old male infant deceased in 1831. Luigi Calamai (1796-1851), a wax craftsman of La Specola workshop in Florence, was commissioned to create a life-sized wax model of the child's head, neck and upper thorax. This artwork allows us to appreciate the cranial and facial alterations determined by 30 lb of cerebrospinal fluid (CSF) accumulated within the cerebral ventricular system. Based on the autopsy report, gross malformations of the neural tube, tumours and haemorrhage could be excluded. A molecular approach proved helpful in confirming sex. We present this case as the so-far most compelling case of hydrocephalus in palaeo-pathological research.


Structured soft particulate matters for delivery of bioactive compounds in foods and functioning in the colon.
The present review discusses challenges, perspectives, and current needs of delivering bioactive compounds (BCs) using soft particulate matters (SPMs) for gut health. SPMs can entrap BCs for incorporation in foods, preserve their bioactivities during processing, storage, and gastrointestinal digestion, and deliver BCs to functioning sites in the colon. To enable these functions, physical, chemical, and biological properties of BCs are integrated in designing various types of SPMs to overcome environmental factors reducing the bioavailability and bioactivity of BCs. The design principles are applied using food grade molecules with the desired properties to produce SPMs by additionally considering the cost, sustainability, and scalability of manufacturing processes. Lastly, to make delivery systems practical, impacts of SPMs on food quality are to be evaluated case by case, and health benefits of functional foods incorporated with delivery systems are to be confirmed and must outweigh the cost of preparing SPMs.


Sensitization to pets is a major determinant of persistent asthma and new asthma onset in Sweden.
Our knowledge about atopy as a longitudinal predictor of asthma is limited. The purpose of this study was to investigate the prognosis of asthma and risk factors for asthma onset, especially sensitization of specific allergens in a population sample. A cohort responded to a respiratory questionnaire in 1990 and 2003. At baseline, 2,060 subjects who, in the screening questionnaire, reported respiratory symptoms and 482 controls were investigated with interviews, spirometry, and skin-prick test. A total of 721 asthmatics and 976 subjects without respiratory disease were clinically verified. At follow-up in 2003, 340 subjects with persistent asthma and 186 subjects with asthma remission were identified, while 76 subjects reported new asthma onset. Sensitization to pets and a high symptom score were significant determinants of persistent asthma (odds ratio (OR) 3.2 (95% CI 1.9-5.6) and 5.7 (2.5-13.3), respectively) and onset of asthma (OR 2.6 (1.1-6.0), and 1.7 (1.2-2.3)). A high self-reported responsiveness to airway irritants (OR 1.6 (1.1-2.2)), and more asthma medications (OR 2.0 (1.3-2.9)) were additional indicators of persistent asthma at the follow-up. Belonging to the older age group decreased the risk both of having persistent asthma and asthma onset. Asthmatics sensitized to pets have a more severe outcome than asthmatics not sensitized to pets. Sensitization to pets was also a strong predictor for onset of asthma. Special attention should be given to asthmatics who report having severe symptoms and problems with airway irritants as such patients are more likely to have persistent problems.


Periovulatory 17 beta-estradiol changes and embryo morphologic features in conception and nonconceptional cycles after human in vitro fertilization.
Thirty-one patients who became pregnant after in vitro fertilization were compared with a matched control group of 31 patients who underwent embryo transfer but did not conceive. During the follicular phase there was no difference in the mean daily estradiol (E2) levels between the pregnant and control groups. In 95% of the women who conceived, E2 continued to rise or plateau on the day after human chorionic gonadotropin (hCG) administration; this pattern was seen in only 62.5% of the control group. There were significantly more "regular" embryos (i.e., those with symmetric blastomeres and no visible cytoplasmic fragments) replaced in patients who became pregnant, compared with the control subjects (65.6% versus 47.0%, P less than 0.05). Embryo regularity was associated with preovulatory E2 pattern; falling E2 levels on the day after hCG administration were associated with transfer of only 18.8% of regular embryos in the control group, whereas in the conceptional cycles with rising E2 levels, 61.5% of the replaced embryos were morphologically regular (P less than 0.01). These results would indicate that changes in the follicular environment as reflected by peripheral E2 levels might affect the developmental potential of the human embryo.


Whiplash: still a pain in the neck.
Whiplash is a common problem, particularly following motor vehicle accidents and may have significant sequelae in terms of disability and financial compensation. Recent research has demonstrated that a number of commonplace medical practices as well as the compensation system may lead to unfavourable outcomes. This article discusses recent research into whiplash and its implications for clinical practice. A full assessment of biopsychosocial factors in the acute phase of the injury is essential to predict those at risk of chronicity. Simple therapeutic and educational measures should be employed and early referral to a psychologist or pain specialist considered for those at high risk.


Do Patients With a Baseline Clinical Condition Warranting the Cautious Use of Parenteral Nutrition Develop Subsequent Metabolic Complications?
The American Society for Parenteral and Enteral Nutrition Adult Nutrition Support Core Curriculum describes clinical conditions that warrant cautious use of parenteral nutrition (CCWCPN). The Core Curriculum authors acknowledge there is no evidence for specific criteria suggested for the clinical conditions. Consequently, the purpose of this study was to determine the impact of a baseline CCWCPN on the development of subsequent metabolic complications in patients receiving parenteral nutrition (PN). Adult patients initiated on PN from May 2014 to July 2015 at Cooper University Hospital were included in this retrospective study. The impact of a CCWCPN on the development of the following was determined: acid-base disturbances, hepatobiliary complications, hypercapnia, hyperchloremia, hyperglycemia, hypernatremia, hypertriglyceridemia, hypochloremia, hypoglycemia, hypokalemia, hypophosphatemia, and refeeding syndrome. Three hundred forty-one patients were included (mean age, 61.7 years; mean duration of PN, 8.5 days; central PN, 97%). Metabolic complications occurred more frequently in patients with a baseline CCWCPN than without these conditions (77% vs 53%, P = .001). Subgroup analyses for the development of metabolic complications in patients with or without each individual baseline CCWCPN yielded the following statistically significant results: hypernatremia (93% vs 57%, P = .007) and hyperchloremia (86% vs 57%, P = .033). Hospitalized adult patients with a baseline CCWCPN were more likely to develop a metabolic complication when receiving PN. Baseline hypernatremia and hyperchloremia were associated with the development of metabolic complications. Baseline CCWCPN should be recognized upon initiation of PN; practitioners should closely monitor patients to minimize subsequent metabolic complications.


Association of testosterone with estrogen abolishes the beneficial effects of estrogen treatment by increasing ROS generation in aorta endothelial cells.
Testosterone has been added to hormone replacement therapy to treat sexual dysfunction in postmenopausal women. Whereas estrogen has been associated with vascular protection, the vascular effects of testosterone are contradictory and the effects of its association with estrogen are largely unknown. In this study we determined the effects of testosterone associated with conjugated equine estrogen (CEE) on vascular function using a model of hypertensive postmenopausal female: ovariectomized spontaneously hypertensive rats. Female spontaneously hypertensive rats were divided into sham-operated, ovariectomized (OVX), and OVX treated for 15 days with either CEE alone (OVX+CEE) or associated with testosterone (OVX+CEE+T). Angiotensin II (ANG II)-induced contraction was markedly increased in aortic rings from OVX compared with sham-operated rats. CEE treatment restored ANG-II responses, a beneficial effect abrogated with CEE+T. CEE treatment also increased endothelium-dependent relaxation, which was impaired in OVX rats. This effect was lost by CEE+T. Treatment of aortas with losartan (ANG-II type-1 receptor antagonist) or apocynin (NADPH-oxidase inhibitor) restored the endothelium-dependent relaxation in OVX and CEE+T, establishing an interplay between ANG-II and endothelial dysfunction in OVX and CEE+T. The benefits by CEE were associated with downregulation of NADPH-oxidase subunits mRNA expression and decreased reactive oxygen species generation. The association of testosterone with CEE impairs the benefits of estrogen on OVX-associated endothelial dysfunction and reactive oxygen species generation in rat aorta by a mechanism that involves phosphorylation of the cytosolic NADPH-oxidase subunit p47(phox).


Emulating a target trial in case-control designs: an application to statins and colorectal cancer.
Previous case-control studies have reported a strong association between statin use and lower cancer risk. It is unclear whether this association reflects a benefit of statins or is the result of design decisions that cannot be mapped to a (hypothetical) target trial (that would answer the question of interest). We outlined the protocol of a target trial to estimate the effect of statins on colorectal cancer incidence among adults with low-density lipoprotein (LDL) cholesterol below 5 mmol/L. We then emulated the target trial using linked electronic health records of 752 469 eligible UK adults (CALIBER 1999-2016) under both a cohort design and a case-control sampling of the cohort. We used pooled logistic regression to estimate intention-to-treat and per-protocol effects of statins on colorectal cancer, with adjustment for baseline and time-varying risk factors via inverse-probability weighting. Finally, we compared our case-control effect estimates with those obtained using previous case-control procedures. Over the 6-year follow-up, 3596 individuals developed colorectal cancer. Estimated intention-to-treat and per-protocol hazard ratios were 1.00 (95% confidence interval [CI]: 0.87, 1.16) and 0.90 (95% CI: 0.71, 1.12), respectively. As expected, adequate case-control sampling yielded the same estimates. By contrast, previous case-control analytical approaches yielded estimates that appeared strongly protective (odds ratio 0.57, 95% CI: 0.36, 0.91, for ≥5 vs. <5 years of statin use). Our study demonstrates how to explicitly emulate a target trial using case-control data to reduce discrepancies between observational and randomized trial evidence. This approach may inform future case-control analyses for comparative effectiveness research.


Antagonistic regulation of Fus2p nuclear localization by pheromone signaling and the cell cycle.
When yeast cells sense mating pheromone, they undergo a characteristic response involving changes in transcription, cell cycle arrest in early G1, and polarization along the pheromone gradient. Cells in G2/M respond to pheromone at the transcriptional level but do not polarize or mate until G1. Fus2p, a key regulator of cell fusion, localizes to the tip of the mating projection during pheromone-induced G1 arrest. Although Fus2p was expressed in G2/M cells after pheromone induction, it accumulated in the nucleus until after cell division. As cells arrested in G1, Fus2p was exported from the nucleus and localized to the nascent tip. Phosphorylation of Fus2p by Fus3p was required for Fus2p export; cyclin/Cdc28p-dependent inhibition of Fus3p during late G1 through S phase was sufficient to block exit. However, during G2/M, when Fus3p was activated by pheromone signaling, Cdc28p activity again blocked Fus2p export. Our results indicate a novel mechanism by which pheromone-induced proteins are regulated during the transition from mitosis to conjugation.


Mixed features in major depressive disorder: diagnoses and treatments.
For the first time in 20 years, the American Psychiatric Association (APA) updated the psychiatric diagnostic system for mood disorders in the Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition (DSM-5). Perhaps one of the most notable changes in the DSM-5 was the recognition of the possibility of mixed symptoms in major depression and related disorders (MDD). While MDD and bipolar and related disorders are now represented by 2 distinct chapters, the addition of a mixed features specifier to MDD represents a structural bridge between bipolar and major depression disorders, and formally recognizes the possibility of a mix of hypomania and depressive symptoms in someone who has never experienced discrete episodes of hypomania or mania. This article reviews historical perspectives on "mixed states" and the recent literature, which proposes a range of approaches to understanding "mixity." We discuss which symptoms were considered for inclusion in the mixed features specifier and which symptoms were excluded. The assumption that mixed symptoms in MDD necessarily predict a future bipolar course in patients with MDD is reviewed. Treatment for patients in a MDD episode with mixed features is critically considered, as are suggestions for future study. Finally, the premise that mood disorders are necessarily a spectrum or a gradient of severity progressing in a linear manner is argued.


A mitochondrial malady: stubborn seizures and atypical migraine.
A 22-year-old female migraineur presented with recurrent convulsive status epilepticus and ataxia. Her epilepsy proved refractory to treatment, necessitating the use of five anti-epileptic drugs and a course of steroids. Genetic testing revealed compound heterozygosity for two mutations of the polymerase-? gene. The case highlights the clinical features and therapeutic challenges associated with this relatively common, but probably underrecognised, mitochondrial disease.


Polarized trafficking of the aquaporin-3 water channel is mediated by an NH2-terminal sorting signal.
Epithelial renal collecting duct cells express multiple types of aquaporin (AQP) water channels in a polarized fashion. AQP2 is specifically targeted to the apical cell domain, whereas AQP3 and AQP4 are expressed on the basolateral membrane. It is crucial that these AQP variants are sorted to their proper polarized membrane domains, because correct AQP sorting enables efficient water transport. However, the molecular mechanisms involved in the polarized targeting and membrane trafficking of AQPs remain largely unknown. In the present study, we have examined the polarized trafficking and surface expression of AQP3 in Madin-Darby canine kidney type II (MDCKII) cells in an effort to identify the molecular determinants of polarized targeting specificity. When expressed in MDCKII cells, the majority of the exogenous wild-type AQP3 was found to be targeted to the basolateral membrane, consistent with its localization pattern in vivo. A potential sorting signal consisting of tyrosine- and dileucine-based motifs was subsequently identified in the AQP3 NH2 terminus. When mutations were introduced into this signaling region, the basolateral targeting of the resulting mutant AQP3 was disrupted and the mutant protein remained in the cytoplasm. AQP2-AQP3 chimeras were then generated in which the entire NH2 terminus of AQP2 was replaced with the AQP3 NH2 terminus. This chimeric protein was observed to be mislocalized constitutively in the basolateral membrane, and mutations in the AQP3 NH2-terminal sorting signal abolished this effect. On the basis of these results, we conclude that an NH2-terminal sorting signal mediates the basolateral targeting of AQP3.