Feasibility of a symptom management intervention for adolescents recovering from a hematopoietic stem cell transplant.
Adolescents undergoing a hematopoietic stem cell transplantation (HSCT) experience a variety of adverse effects and eating difficulties. Few interventions exist to assist patients with self-care after HSCT hospitalization. The Eating After Transplant (EAT!) program is a mobile phone applications developed to assist adolescents with self-management of common eating-related issues during HSCT recovery. This study examined the acceptability and usability of the EAT! program among adolescents and assessed the competency of the participants using the program after hospital discharge through the first 100 days after HSCT. A repeated-measures design was used to evaluate the EAT! application with 16 adolescent patients recovering from an allogeneic HSCT. Participants provided verbal feedback and used a Likert scale to rate acceptability and usability of the application. In addition, a tracking device monitored use of the application. Competency was measured with orientation time and independent demonstration of use of the application. Acceptability remained high throughout the study, but use significantly decreased over time. Patients reported familiarity with the program's content as the reason for the declining use. Competency was excellent with a short orientation period and independent demonstration throughout the study. A mobile phone application is a feasible intervention to educate adolescents with symptom management strategies. Future research needs to examine factors affecting sustainability of use over time. Healthcare providers need to continue to develop and evaluate innovative methods to educate adolescents on effective self-care strategies throughout HSCT recovery.


Occipital foramina development involves localised regulation of mesenchyme proliferation and is independent of apoptosis.
Cranial foramina are holes within the skull, formed during development, allowing entry and exit of blood vessels and nerves. Once formed they must remain open, due to the vital structures they contain, i.e. optic nerves, jugular vein, carotid artery, and other cranial nerves and blood vessels. Understanding cranial foramina development is essential as cranial malformations lead to the stenosis or complete closure of these structures, resulting in blindness, deafness, facial paralysis, raised intracranial pressure and lethality. Here we focus on describing early events in the formation of the jugular, carotid and hypoglossal cranial foramina that form in the mesoderm-derived, endochondral occipital bones at the base of the embryonic chick skull. Whole-mount skeletal staining of skulls indicates the appearance of these foramina from HH32/D7.5 onwards. Haematoxylin & eosin staining of sections shows that the intimately associated mesenchyme, neighbouring the contents of these cranial foramina, is initially very dense and gradually becomes sparser as development proceeds. Histological examination also revealed that these foramina initially contain relatively large-diameter nerves, which later become refined, and are closely associated with the blood vessel, which they also innervate within the confines of the foramina. Interestingly cranial foramina in the base of the skull contain blood vessels lacking smooth muscle actin, which suggests these blood vessels belong to glomus body structures within the foramina. The blood vessel shape also appears to dictate the overall shape of the resulting foramina. We initially hypothesised that cranial foramina development could involve targeted proliferation and local apoptosis to cause 'mesenchymal clearing' and the creation of cavities in a mechanism similar to joint cavitation. We find that this is not the case, and propose that a mechanism reliant upon local nerve/blood vessel-derived restriction of ossification may contribute to foramina formation during cranial development.


Circulating osteocrin stimulates bone growth by limiting C-type natriuretic peptide clearance.
Although peptides are safe and useful as therapeutics, they are often easily degraded or metabolized. Dampening the clearance system for peptide ligands is a promising strategy for increasing the efficacy of peptide therapies. Natriuretic peptide receptor B (NPR-B) and its naturally occurring ligand, C-type natriuretic peptide (CNP), are potent stimulators of endochondral bone growth, and activating the CNP/NPR-B system is expected to be a powerful strategy for treating impaired skeletal growth. CNP is cleared by natriuretic peptide clearance receptor (NPR-C); therefore, we investigated the effect of reducing the rate of CNP clearance on skeletal growth by limiting the interaction between CNP and NPR-C. Specifically, we generated transgenic mice with increased circulating levels of osteocrin (OSTN) protein, a natural NPR-C ligand without natriuretic activity, and observed a dose-dependent skeletal overgrowth phenotype in these animals. Skeletal overgrowth in OSTN-transgenic mice was diminished in either CNP- or NPR-C-depleted backgrounds, confirming that CNP and NPR-C are indispensable for the bone growth-stimulating effect of OSTN. Interestingly, double-transgenic mice of CNP and OSTN had even higher levels of circulating CNP and additional increases in bone length, as compared with mice with elevated CNP alone. Together, these results support OSTN administration as an adjuvant agent for CNP therapy and provide a potential therapeutic approach for diseases with impaired skeletal growth.


Early-onset liver cancer in South America associates with low hepatitis B virus DNA burden.
In Peru, hepatocellular carcinoma (HCC) arises in young non-cirrhotic patients. Hepatitis B virus (HBV) is suspected to be the prominent etiological agent. We thus performed a comprehensive molecular study of HBV infection in 65 Peruvian HCC patients. Only 51% were considered as persistently infected at the onset. HBV DNA was found by PCR in the tumor and/or matched non-tumor liver tissues in more than 80% of cases (n = 53/65). HBV DNA was significantly more abundant in livers of younger patients than in those of the older ones. We consistently observed low viral DNA burden (0.1-6.5 copies for 100 cells), with viral genomes in younger patients displaying higher proportion of mutations at di-pyrimidines (TpT and CpC, P = 0.006). A drastic activation of multiple DNA repair pathways in tumors of younger patients was observed. Our observations clearly challenge the current vision that associates high HBV DNA load with earlier tumor development. We concluded that in Peru, and maybe in other populations with Americas' indigenous ancestry, HBV-associated liver tumorigenesis might differ significantly from that generally observed in the rest of the world. Procedures used to screen for HCC development in subjects at risk should be adapted to the local situation.


Academic staff recruitment and retention challenges at the University of Botswana medical school.
Sub-Saharan Africa has a greater share of the global burden of disease, poverty, and inadequate human resources for health compared with other regions of the world. Botswana, as other regional countries, is failing to successfully recruit and retain academics at its medical school. To document the medical school's staff recruitment and retention trends and challenges, and to propose possible solutions. This was a descriptive research study involving review and analysis of the University of Botswana medical school's staff number targets, actual numbers on post, and other relevant publicly available university documents. The numbers and country of origin of staff recruited from 2008 to 2013 were recorded. Net staff gain or loss per year was then calculated. Student numbers were analysed and related to staff availability. As there was a multilevel change in university management in 2011, the periods and events before and after April 2011 were analysed. Publicly available University of Botswana documents about the university's organisational structure, policies, and processes were reviewed. Over a 5-year period, the school recruited 74 academics worldwide; 30 of them left the school. Retention was a greater challenge than recruitment. The school had difficulty recruiting locals and senior academics, regardless of specialty. It appears that staff loss occurred regardless of country of origin. The authors suggest that multilevel change in management was one of the most likely contributors to the school's recruitment and retention challenges. The University of Botswana must comprehensively address these.


De-escalation and limit-setting in forensic mental health units.
Limit-setting and de-escalation are commonly used nursing interventions that are critical to the effective maintenance of the therapeutic milieu and the prevention and management of aggression in forensic mental health units. However, despite their purported importance, the techniques used in these interventions are rarely described. Further, these techniques are seldom based on empirical research or derived from a theory or model of aggression or interpersonal behavior. The purpose of this review is to examine the extant literature on limit-setting and de-escalation and consider how these activities could be enhanced. In concluding, this review emphasizes the potential benefits of drawing upon (1) a model of aggressive behavior and (2) a theory and literature examining interpersonal behavior, to enhance limit-setting and de-escalation skills.


Functional coupling constrains craniofacial diversification in Lake Tanganyika cichlids.
Functional coupling, where a single morphological trait performs multiple functions, is a universal feature of organismal design. Theory suggests that functional coupling may constrain the rate of phenotypic evolution, yet empirical tests of this hypothesis are rare. In fish, the evolutionary transition from guarding the eggs on a sandy/rocky substrate (i.e. substrate guarding) to mouthbrooding introduces a novel function to the craniofacial system and offers an ideal opportunity to test the functional coupling hypothesis. Using a combination of geometric morphometrics and a recently developed phylogenetic comparative method, we found that head morphology evolution was 43% faster in substrate guarding species than in mouthbrooding species. Furthermore, for species in which females were solely responsible for mouthbrooding the males had a higher rate of head morphology evolution than in those with bi-parental mouthbrooding. Our results support the hypothesis that adaptations resulting in functional coupling constrain phenotypic evolution.


A novel optical biosensor for direct and selective determination of serotonin in serum by Solid Surface-Room Temperature Phosphorescence.
This paper describes a novel biosensor which combines the use of nanotechnology (non-woven nanofibre mat) with Solid Surface-Room Temperature Phosphorescence (SS-RTP) measurement for the determination of serotonin in human serum. The developed biosensor is simple and can be directly applied in serum; only requires a simple clean-up protocol. Therefore it is the first time that serotonin is analysed directly in serum with a non-enzymatic technique. This new approach is based on the covalent immobilization of serotonin directly from serum on a functional nanofibre material (Tiss®-Link) with a preactivated surface for direct covalent immobilization of primary and secondary amines, and the subsequent measurement of serotonin phosphorescent emission from the solid surface. The phosphorescent detection allows avoiding the interference from any fluorescence emission or scattering light from any molecule present in the serum sample which can be also immobilised on the nanofibre material. The determination of serotonin with this SS-RTP sensor overcomes some limitations, such as large interference from the matrix and high cost and complexity of many of the methods widely used for serotonin analysis. The potential applicability of the sensor in the clinical diagnosis was demonstrated by analysing serum samples from seven healthy volunteers. The method was validated with an external reference laboratory, obtaining a correlation coefficient of 0.997 which indicates excellent correlation between the two methods.


Isolated Intrapulmonary Vascular Dilatations and the Risk of Developing Hepatopulmonary Syndrome in Liver Transplant Candidates.
The natural history of intrapulmonary vascular dilations (IPVD) and their impact on patient outcomes in the setting of portal hypertension has only been described in small series. To assess the development of hepatopulmonary síndrome (HPS) in patients with isolated IPVD and to evaluate outcomes of IPVD and HPS among patients evaluated for liver transplantation (LT). Data from a prospective cohort of patients evaluated for LT with standardized screening for HPS were analyzed. IPVDs were defined as the presence of microbubbles in the left atrium > 3 cycles following right atrial opacification. HPS was defined as the presence of IPVD and hypoxemia (Alveolar-arterial gradient ≥ 15 mmHg) in the absence of concomitant cardiopulmonary disease. A total of 104 patients with negative contrast-enhanced echocardiogram (CE) were compared to 63 patients with IPVD and 63 patients with HPS. Only four patients were categorized as 'severe' HPS based on degree of hipoxemia (defined as PaO2 < 60 mmHg). Twenty IPVD patients were followed with ABG over a mean duration of 21 months (range 9-43), of whom 7 (35%) subsequently met HPS criteria. Overall unadjusted survival from the time of LT evaluation using multi-state survival models that accounted for pre- and post-LT time was not statistically different among the three groups (negative CE, IPVD, and HPS; p > 0.5). Patients with IPVD appear to have a substantial risk of developing oxygenation impairment over time and progress to HPS. In our cohort, survival in patients with HPS and isolated IPVD is not different when compared to those without IPVDs.


Phage type conversion in Salmonella enterica serotype Enteritidis caused by the introduction of a resistance plasmid of incompatibility group X (IncX).
The plasmid pOG670, a 54 kb, conjugative plasmid that specifies resistance to ampicillin and kanamycin and belonging to the incompatibility group X (IncX), was transferred into 10 isolates of Salmonella enterica serotype Enteritidis belonging to 10 different phage types (PT1, 2, 3, 4, 8, 9, 9b, 10, 11 and 13). Acquisition of the plasmid by these strains did not result in the loss of any resident plasmids but resulted in phage type conversion in 8 of the 10 strains (PT1, 2, 4, 8, 9, 9b, 10 and 11). The observed changes in phage type were found to result from the loss of sensitivity to 3 of the 10 typing phages used (phages 3, 5 and 7). Where the conversion resulted in a change to a defined phage type, both the new and original PTs belonged to the same, previously described, evolutionary lines. Enteritidis PTs 1, 4 and 8, commonly associated with poultry world-wide, were converted to PTs 21, 6 and 13a respectively. The results indicate a different route for phage type conversion Enteritidis from others reported in the literature and, although IncX plasmids are not normally present in PT8 or PT13a, may suggest a possible mechanism/link connecting these phage types.


Phase II study of Amrubicin combined with carboplatin for thymic carcinoma and invasive thymoma: North Japan Lung Cancer group study 0803.
There has been no standard chemotherapy for advanced or recurrent thymic malignancies including thymic carcinoma (TC) and invasive thymoma (IT), though platinum and anthracycline have been reported as effective agents for the treatment of these diseases. The objective of this study was to evaluate the efficacy and safety of the combination of amrubicin (AMR), a new anthracycline agent, and carboplatin (CBDCA) in patients with advanced thymic malignancies. Patients with histologically confirmed thymic malignancies received AMR (35 mg/m, days 1-3) and CBDCA (area under the curve 4.0, day 1) every 3 weeks. Patients who had received previous chemotherapy were treated with a reduced dose of AMR (30 mg/m). The primary end point was objective response rate (ORR), and secondary endpoints were progression-free survival, overall survival, and toxicity profile. From December 2008 to October 2012, 51 patients (33 TC and 18 IT) were enrolled. The median number of treatment cycles was four in each group. The ORR and progression-free survival were 30% (95% confidence interval, 14-46) and 7.6 months in the TC group, and 17% (95% confidence interval, 0-34) and 7.6 months in the IT group, respectively. The ORR of TC patients without previous chemotherapy (n = 19) was 42%. Although grade 3 or 4 hematological toxicities were common including neutropenia (82%) and febrile neutropenia (22%), these were transient and manageable. Nonhematological toxicities were moderate and no treatment-related death was observed. The combination of AMR with CBDCA was active for TC with acceptable toxicity, although it was not effective for IT. Further investigation of this regimen for advanced TC is warranted.


Cadmium-induced effects on cellular signaling pathways in the liver of transgenic estrogen reporter mice.
Estrogen-like effects of cadmium (Cd) have been reported in several animal studies, and recent epidemiological findings suggest increased risk of hormone-dependent cancers after Cd exposure. The mechanisms underlying these effects are still under investigation. Our aim was to study the effects of Cd on cellular signaling pathways in vivo with special focus on estrogen signaling and to perform benchmark dose analysis on the effects. Transgenic adult ERE-luciferase male mice were exposed subcutaneously to 0.5-500 μg CdCl(2) per kg body weight (bw) or 17α-ethinylestradiol (EE2) for 3 days. These doses had no effects on organ and bw or testicular histology, indicating subtoxic exposure levels. The transgene luciferase, reporting genomic estrogen response, was significantly increased by EE2 but not by Cd. However, Cd significantly affected kinase phosphorylation and endogenous gene expression. Interestingly, gene expression changes displayed a traditional dose-response relationship, with benchmark dose levels for the expression of Mt1, Mt2, p53, c-fos, and Mdm2 being 92.9, 19.9, 7.6, 259, and 25.9 μg/kg bw, respectively, but changes in kinase phosphorylation were only detected at low exposure levels. Phosphorylation of Erk1/2 was significantly increased even in the lowest dose group, 0.5 μg/kg bw, rendering pErk1/2 a more sensitive sensor of exposure than changes in gene expression. Collectively, our data suggest that the effects triggered by Cd in vivo are markedly concentration dependent. Furthermore, we conclude that the estrogen-like effects of Cd are likely to result from a mechanism different from steroidal estrogens.


Nausea and vomiting in early pregnancy: Effects on food intake and diet quality.
Experiences of nausea and/or vomiting in pregnancy (NVP) vary greatly, but the paucity of studies with pre-pregnancy dietary data mean that little is known about the effects of NVP on diet. Using an administered food frequency questionnaire, diet was assessed before pregnancy and at 11 and 34 weeks' gestation in 2270 participants in a UK birth cohort study (Southampton Women's Survey). Experience of NVP in early pregnancy was graded as none, mild, moderate, or severe. Participants reported their level of food consumption as more, the same, or less than before pregnancy. "Prudent" diet scores (derived using principal component analysis) were used to describe participants' diet quality before, in early and late pregnancy. In early pregnancy, 89% of women were nauseous, although most commonly, the NVP experienced was mild (48%) or moderate (30%); 11% had severe NVP. A total of 39% of women reported an increase in their level of food intake in early pregnancy; 34% reported a reduction. Increasing severity of nausea was associated with changes in intake of a range of foods, most notably reduced consumption of vegetables, tea/coffee, rice/pasta, breakfast cereals, beans/pulses and citrus fruits/fruit juices and increased consumption of white bread, and soft drinks. Increasing severity of nausea was also associated with decreasing prudent diet score from before to early pregnancy, such that women with severe nausea had prudent diet scores 0.29 SDs lower than those with no nausea (P < 0.001). However, this was transient as NVP was not related to change in diet quality from before to late pregnancy.


Development of experimental autoimmune uveitis: efficient recruitment of monocytes is independent of CCR2.
Macrophages are major contributors to the damage occurring in the retina in experimental autoimmune uveitis (EAU). CCR2 may be needed for efficient recruitment of monocytes to an inflammatory site, and the aim of this study was to determine whether this was the case in EAU. EAU was induced and graded in C57BL/6J and CCR2(-/-) mice. Macrophage infiltration and CCR2 expression were assessed using immunohistochemistry. Retinas were examined for MCP-1 expression using RT-PCR. Rolling and infiltration of labeled bone marrow monocytes at the inflamed retinal vasculature were examined by scanning laser ophthalmoscopy and confocal microscopy, respectively. Effect of CCR2 deletion or blockade by antibody and antagonist was determined. Expression of mRNA for MCP-1 increased as EAU developed and was localized to the retina. CCR2 was associated with infiltrating macrophages. However, EAU induced in CCR2(-/-) mice was not reduced in severity, and neither was the percentage of macrophages in the retina. CCR2(-/-) monocytes, 48 hours after adoptive transfer to mice with EAU, showed no significant difference in percentage rolling or infiltration into the retina compared to WT. CCR2-independent rolling of monocytes was confirmed by CCR2 neutralizing antibody and antagonist treatment. CCR2 does not have a primary role in the recruitment of monocytes to the inflammatory site across the blood-retina barrier in well-developed EAU. Therapeutics targeting CCR2 are unlikely to be of value in treating human posterior uveitis.


A novel laser-Doppler flowmetry assisted murine model of acute hindlimb ischemia-reperfusion for free flap research.
Suitable and reproducible experimental models of translational research in reconstructive surgery that allow in-vivo investigation of diverse molecular and cellular mechanisms are still limited. To this end we created a novel murine model of acute hindlimb ischemia-reperfusion to mimic a microsurgical free flap procedure. Thirty-six C57BL6 mice (n = 6/group) were assigned to one control and five experimental groups (subject to 6, 12, 96, 120 hours and 14 days of reperfusion, respectively) following 4 hours of complete hindlimb ischemia. Ischemia and reperfusion were monitored using Laser-Doppler Flowmetry. Hindlimb tissue components (skin and muscle) were investigated using histopathology, quantitative immunohistochemistry and immunofluorescence. Despite massive initial tissue damage induced by ischemia-reperfusion injury, the structure of the skin component was restored after 96 hours. During the same time, muscle cells were replaced by young myotubes. In addition, initial neuromuscular dysfunction, edema and swelling resolved by day 4. After two weeks, no functional or neuromuscular deficits were detectable. Furthermore, upregulation of VEGF and tissue infiltration with CD34-positive stem cells led to new capillary formation, which peaked with significantly higher values after two weeks. These data indicate that our model is suitable to investigate cellular and molecular tissue alterations from ischemia-reperfusion such as occur during free flap procedures.


Registration of in-vivo to ex-vivo MRI of surgically resected specimens: a pipeline for histology to in-vivo registration.
Advances in MRI have the potential to improve surgical treatment of epilepsy through improved identification and delineation of lesions. However, validation is currently needed to investigate histopathological correlates of these new imaging techniques. The purpose of this work is to develop and evaluate a protocol for deformable image registration of in-vivo to ex-vivo resected brain specimen MRI. This protocol, in conjunction with our previous work on ex-vivo to histology registration, completes a registration pipeline for histology to in-vivo MRI, enabling voxel-based validation of novel and existing MRI techniques with histopathology. A combination of image-based and landmark-based 3D registration was used to register in-vivo MRI and the ex-vivo MRI from patients (N=10) undergoing epilepsy surgery. Target registration error (TRE) was used to assess accuracy and the added benefit of deformable registration. A mean TRE of 1.35±0.11 and 1.41±0.33mm was found for neocortical and hippocampal specimens respectively. Statistical analysis confirmed that the deformable registration significantly improved the registration accuracy for both specimens. Image registration of surgically resected brain specimens is a unique application which presents numerous technical challenges and that have not been fully addressed in previous literature. Our computed TRE are comparable to previous attempts tackling similar applications, as registering in-vivo MRI to whole brain or serial histology. The presented registration pipeline finds dense and accurate spatial correspondence between in-vivo MRI and histology and allows for the spatially local and quantitative assessment of pathological correlates in MRI.


Report of the WHO technical consultation on the effect of maternal influenza and influenza vaccination on the developing fetus: Montreal, Canada, September 30-October 1, 2015.
In 2012, the World Health Organization (WHO) released a position paper on influenza vaccination recommending that pregnant women have the highest priority for seasonal vaccination in countries where the initiation or expansion of influenza immunization programs is under consideration. Although the primary goal of the WHO recommendation is to prevent influenza illness in pregnant women, the potential benefits of maternal immunization in protecting young infants are also recognized. The extent to which maternal influenza vaccination may prevent adverse birth outcomes such as preterm birth or small-for-gestational-age birth, however, is unclear as available studies are in disagreement. To inform WHO about the empirical evidence relating to possible benefits of influenza vaccination on birth outcomes, a consultation of experts was held in Montreal, Canada, September 30-October 1, 2015. Presentations and discussions covered a broad range of issues, including influenza virus infection during pregnancy and its effect on the health of the mother and the fetus, possible biological mechanisms for adverse birth outcomes following maternal influenza illness, evidence on birth outcomes following influenza illness during pregnancy, evidence from both observational studies and randomized controlled trials on birth outcomes following influenza vaccination of pregnant women, and methodological issues. This report provides an overview of the presentations, discussions and conclusions.


The feasibility of creating a checklist for the assessment of the methodological quality both of randomised and non-randomised studies of health care interventions.
To test the feasibility of creating a valid and reliable checklist with the following features: appropriate for assessing both randomised and non-randomised studies; provision of both an overall score for study quality and a profile of scores not only for the quality of reporting, internal validity (bias and confounding) and power, but also for external validity. A pilot version was first developed, based on epidemiological principles, reviews, and existing checklists for randomised studies. Face and content validity were assessed by three experienced reviewers and reliability was determined using two raters assessing 10 randomised and 10 non-randomised studies. Using different raters, the checklist was revised and tested for internal consistency (Kuder-Richardson 20), test-retest and inter-rater reliability (Spearman correlation coefficient and sign rank test; kappa statistics), criterion validity, and respondent burden. The performance of the checklist improved considerably after revision of a pilot version. The Quality Index had high internal consistency (KR-20: 0.89) as did the subscales apart from external validity (KR-20: 0.54). Test-retest (r 0.88) and inter-rater (r 0.75) reliability of the Quality Index were good. Reliability of the subscales varied from good (bias) to poor (external validity). The Quality Index correlated highly with an existing, established instrument for assessing randomised studies (r 0.90). There was little difference between its performance with non-randomised and with randomised studies. Raters took about 20 minutes to assess each paper (range 10 to 45 minutes). This study has shown that it is feasible to develop a checklist that can be used to assess the methodological quality not only of randomised controlled trials but also non-randomised studies. It has also shown that it is possible to produce a checklist that provides a profile of the paper, alerting reviewers to its particular methodological strengths and weaknesses. Further work is required to improve the checklist and the training of raters in the assessment of external validity.


Mortality, mauling, and maiming by vicious dogs.
Maiming and death due to dog bites are uncommon but preventable tragedies. We postulated that patients admitted to a level I trauma center with dog bites would have severe injuries and that the gravest injuries would be those caused by pit bulls. We reviewed the medical records of patients admitted to our level I trauma center with dog bites during a 15-year period. We determined the demographic characteristics of the patients, their outcomes, and the breed and characteristics of the dogs that caused the injuries. Our Trauma and Emergency Surgery Services treated 228 patients with dog bite injuries; for 82 of those patients, the breed of dog involved was recorded (29 were injured by pit bulls). Compared with attacks by other breeds of dogs, attacks by pit bulls were associated with a higher median Injury Severity Scale score (4 vs. 1; P = 0.002), a higher risk of an admission Glasgow Coma Scale score of 8 or lower (17.2% vs. 0%; P = 0.006), higher median hospital charges ($10,500 vs. $7200; P = 0.003), and a higher risk of death (10.3% vs. 0%; P = 0.041). Attacks by pit bulls are associated with higher morbidity rates, higher hospital charges, and a higher risk of death than are attacks by other breeds of dogs. Strict regulation of pit bulls may substantially reduce the US mortality rates related to dog bites.


Discovery of candidate disease genes in ENU-induced mouse mutants by large-scale sequencing, including a splice-site mutation in nucleoredoxin.
An accurate and precisely annotated genome assembly is a fundamental requirement for functional genomic analysis. Here, the complete DNA sequence and gene annotation of mouse Chromosome 11 was used to test the efficacy of large-scale sequencing for mutation identification. We re-sequenced the 14,000 annotated exons and boundaries from over 900 genes in 41 recessive mutant mouse lines that were isolated in an N-ethyl-N-nitrosourea (ENU) mutation screen targeted to mouse Chromosome 11. Fifty-nine sequence variants were identified in 55 genes from 31 mutant lines. 39% of the lesions lie in coding sequences and create primarily missense mutations. The other 61% lie in noncoding regions, many of them in highly conserved sequences. A lesion in the perinatal lethal line l11Jus13 alters a consensus splice site of nucleoredoxin (Nxn), inserting 10 amino acids into the resulting protein. We conclude that point mutations can be accurately and sensitively recovered by large-scale sequencing, and that conserved noncoding regions should be included for disease mutation identification. Only seven of the candidate genes we report have been previously targeted by mutation in mice or rats, showing that despite ongoing efforts to functionally annotate genes in the mammalian genome, an enormous gap remains between phenotype and function. Our data show that the classical positional mapping approach of disease mutation identification can be extended to large target regions using high-throughput sequencing.


Screening for dementia by telephone using the memory impairment screen.
: To develop and assess telephone-based screening tests for dementia, especially Alzheimer's disease (AD). : A cross-sectional validation study nested within a longitudinal study of aging and dementia. : The Einstein Aging Study of the Albert Einstein College of Medicine, Bronx, New York. : Three hundred elderly community volunteers living in Bronx County, 27 of whom were diagnosed with dementia based on in-person clinical evaluation. Of the 27 individuals with dementia, 18 had AD. : A telephone battery was administered that included the Memory Impairment Screen by telephone (MIS-T, a test of semantic memory), the Category Fluency Test (CF-T), and the Telephone Instrument for Cognitive Status (TICS). An in-person evaluation then followed that included a neurological examination, a neuropsychological battery, demographics, and medical history. : The telephone battery was well accepted. The MIS-T required 4 minutes; the CF-T, 3 minutes; and the TICS, 10 minutes. The MIS-T had excellent sensitivity and specificity when compared with the CF-T and the TICS. Using cutscores on all three tests that provide a sensitivity of 78%, specificity was significantly higher for the MIS-T (93%) than for the CF-T (78%, P<.05) or the TICS (80%, P<.05). Combining the MIS-T and CF-T improved discriminative validity but increased screening time and the complexity of scoring. Normative data for the MIS-T, the CF-T, and the TICS for use in settings with different base rates (prevalence) of dementia are presented in this study. : The MIS-T outperforms the CF-T and the TICS as a valid and time-efficient telephone screen for dementia. For applications that require optimal efficiency and accuracy, the MIS-T is recommended.


Clinical and histopathological characteristics of patients with prostate cancer in the BioBank Japan project.
Prostate cancer is the sixth leading cause of cancer-related deaths in Japan. We aimed to elucidate the clinical and histopathological characteristics of patients with prostate cancer in the BioBank Japan (BBJ) project. Four thousand, seven hundred and ninety-three patients diagnosed with prostate cancer in the BBJ project were included. Clinical and histopathological data, including causes of death, were analyzed. Relative survival (RS) rates of prostate cancer were calculated. Four thousand, one hundred and seventy-one prostate cancer patients with available histological data had adenocarcinoma. The mean age of the patients was 72.5 years. The proportion of patients who were non-smokers, non-drinkers, had a normal body mass index, did not exercise, had a normal prostate-specific antigen level, and had a family history of prostate cancer were 30.7%, 28.0%, 66.6%, 58.1%, 67.6%, and 6.5%, respectively. The proportion of patients with Stage II, III, and IV disease were 24.4%, 7.3%, and 4.4%, respectively. After limiting to patients with a time from the initial diagnosis of prostate cancer to entry into the study cohort of ≤90 days (n = 869), the 5- and 10-year RS rates were 96.3% and 100.5%, respectively, although we were unable to consider management strategies due to a plenty of data missing. We provide an overview of patients with prostate cancer in the BBJ project. Our findings, coupled with those from various high throughput "omics" technologies, will contribute to the implementation of prevention interventions and medical management of prostate cancer patients.


Relation between oral health and nutritional condition in the elderly.
Oral health is a prerequisite for a good chewing function, which may have an impact on food choices and nutritional well-being. This study was designed to evaluate the relationship between oral health status and nutritional status in the elderly. In this cross-sectional study, 33 elderly people from the Group for the Elderly Interdisciplinary Geriatrics and Gerontology Program, at Fluminense Federal University, Niteroi, RJ, Brazil, completed a questionnaire to collect information on socioeconomic status, eating habits, physical activity and health habits, undertook a clinical oral examination, blood test, and anthropometric measurements, and were allocated into groups according to age. The oral health status was assessed using the index for decayed, missing and filled teeth (DMFT). The nutritional status was assessed using hemoglobin, hematocrit and albumin concentrations in blood, anthropometric values and the body mass index. Tooth loss was the biggest nuisance to the elderly subjects (57.6%), followed by the use of dentures (30.3%) and ill-fitting dentures (33.3%). 66.6% of patients had difficulty in chewing, and 54.5% reported this to be due to prostheses and 13.6% to the absence of teeth. A significant correlation was found between DMFT and the value of suprailiac skinfold thickness (rho=0.380, p=0.029). The results support the temporal association between tooth loss and detrimental changes in anthropometry, which could contribute to increased risk of developing chronic diseases.


Remote-sensing infrared thermometer with radiation balancing.
What is to our knowledge a novel infrared thermometer (IRT) for remote measurement of the temperature rise (-20-100 K) above the variable ambient (270-320 K) of a distant object is described. A radiation-balancing method is successfully extended to the near-ambient temperature range by variation of the temperature of a built-in blackbody, until the radiation from it equals the radiation from the source, so that its temperature is proportional to that of the source. Another feature believed to be novel is simplifying the design by elimination of the need for cooling the blackbody for subambient temperature range by use of a second blackbody, strategically located, which is heated to achieve radiation balance. Detailed theoretical analysis is given, showing that the IRT can measure remotely the total emissivity or even the electric current or voltage. Resistive inserts are proposed for improving the accuracy of current measurement. A method is proposed for simultaneous remote measurement of absolute temperature and emissivity by variation of the heating current and the aperture of the blackbody for radiation balancing in two bands so that prior knowledge of the object's emissivity is not needed.


Are the three hydroxyphenyl radical isomers created equal?--The role of the phenoxy radical.
We have investigated the thermal decomposition of the three hydroxyphenyl radicals (˙C6H4OH) in a heated microtubular reactor. Intermediates and products were identified isomer-selectively applying photoion mass-selected threshold photoelectron spectroscopy with vacuum ultraviolet synchrotron radiation. Similarly to the phenoxy radical (C6H5-O˙), hydroxyphenyl decomposition yields cyclopentadienyl (c-C5H5) radicals in a decarbonylation reaction at elevated temperatures. This finding suggests that all hydroxyphenyl isomers first rearrange to form phenoxy species, which subsequently decarbonylate, a mechanism which we also investigate computationally. Meta- and para-radicals were selectively produced and spectroscopically detectable, whereas the ortho isomer could not be traced due to its fast rethermalization and rapid decomposition in the reactor. A smaller barrier to isomerization to phenoxy was found to be the reason for this observation. Since hydroxyphenyl species may be present under typical sooting conditions in flames, the resonantly stabilized cyclopentadienyl radical adds to the hydrocarbon pool and can contribute to the formation of polycyclic aromatic hydrocarbons, which are precursors in soot formation.


Relation between rumination and impaired memory in dysphoric moods.
College students in dysphoric or nondysphoric moods studied pairs of words and later took a fragment-completion test of memory for targets from the pairs (under process-dissociation procedures for obtaining estimates of controlled and automatic retrieval; L. L. Jacoby, 1996). Between the study and test phases, some participants waited quietly for 7 min; others rated self-focused materials designed to invoke ruminations in the dysphoric group; and still others rated self-irrelevant and task-irrelevant materials. A dysphoria-related impairment in controlled retrieval occurred in the first 2 conditions but not in the 3rd condition. These results show that the nature of task-irrelevant thoughts contributes to memory impairments in dysphoria and suggest that self-focused rumination might also contribute to similar impairments under unconstrained conditions that permit mind wandering.


Microhomology-mediated end joining is activated in irradiated human cells due to phosphorylation-dependent formation of the XRCC1 repair complex.
Microhomology-mediated end joining (MMEJ), an error-prone pathway for DNA double-strand break (DSB) repair, is implicated in genomic rearrangement and oncogenic transformation; however, its contribution to repair of radiation-induced DSBs has not been characterized. We used recircularization of a linearized plasmid with 3΄-P-blocked termini, mimicking those at X-ray-induced strand breaks, to recapitulate DSB repair via MMEJ or nonhomologous end-joining (NHEJ). Sequence analysis of the circularized plasmids allowed measurement of relative activity of MMEJ versus NHEJ. While we predictably observed NHEJ to be the predominant pathway for DSB repair in our assay, MMEJ was significantly enhanced in preirradiated cells, independent of their radiation-induced arrest in the G2/M phase. MMEJ activation was dependent on XRCC1 phosphorylation by casein kinase 2 (CK2), enhancing XRCC1's interaction with the end resection enzymes MRE11 and CtIP. Both endonuclease and exonuclease activities of MRE11 were required for MMEJ, as has been observed for homology-directed DSB repair (HDR). Furthermore, the XRCC1 co-immunoprecipitate complex (IP) displayed MMEJ activity in vitro, which was significantly elevated after irradiation. Our studies thus suggest that radiation-mediated enhancement of MMEJ in cells surviving radiation therapy may contribute to their radioresistance and could be therapeutically targeted.


'Particle genetics': treating every cell as unique.
Genotype-phenotype relations are usually inferred from a deterministic point of view. For example, quantitative trait loci (QTL), which describe regions of the genome associated with a particular phenotype, are based on a mean trait difference between genotype categories. However, living systems comprise huge numbers of cells (the 'particles' of biology). Each cell can exhibit substantial phenotypic individuality, which can have dramatic consequences at the organismal level. Now, with technology capable of interrogating individual cells, it is time to consider how genotypes shape the probability laws of single cell traits. The possibility of mapping single cell probabilistic trait loci (PTL), which link genomic regions to probabilities of cellular traits, is a promising step in this direction. This approach requires thinking about phenotypes in probabilistic terms, a concept that statistical physicists have been applying to particles for a century. Here, I describe PTL and discuss their potential to enlarge our understanding of genotype-phenotype relations.


mTOR is a fine tuning molecule in CDK inhibitors-induced distinct cell death mechanisms via PI3K/AKT/mTOR signaling axis in prostate cancer cells.
Purvalanol and roscovitine are cyclin dependent kinase (CDK) inhibitors that induce cell cycle arrest and apoptosis in various cancer cells. We further hypothesized that co-treatment of CDK inhibitors with rapamycin, an mTOR inhibitor, would be an effective combinatory strategy for the inhibition of prostate cancer regard to androgen receptor (AR) status due to inhibition of proliferative pathway, PI3K/AKT/mTOR, and induction of cell death mechanisms. Androgen responsive (AR+), PTEN(-/-) LNCaP and androgen independent (AR-), PTEN(+/-) DU145 prostate cancer cells were exposed to purvalanol (20 µM) and roscovitine (30 µM) with or without rapamycin for 24 h. Cell viability assay, immunoblotting, flow cytometry and fluorescence microscopy was used to define the effect of CDK inhibitors with or without rapamycin on proliferative pathway and cell death mechanisms in LNCaP and DU145 prostate cancer cells. Co-treatment of rapamycin modulated CDK inhibitors-induced cytotoxicity and apoptosis that CDK inhibitors were more potent to induce cell death in AR (+) LNCaP cells than AR (-) DU145 cells. CDK inhibitors in the presence or absence of rapamycin induced cell death via modulating upstream PI3K/AKT/mTOR signaling pathway in LNCaP cells, exclusively only treatment of purvalanol have strong potential to inhibit both upstream and downstream targets of mTOR in LNCaP and DU145 cells. However, co-treatment of rapamycin with CDK inhibitors protects DU145 cells from apoptosis via induction of autophagy mechanism. We confirmed that purvalanol and roscovitine were strong apoptotic and autophagy inducers that based on regulation of PI3K/AKT/mTOR signaling pathway. Co-treatment of rapamycin with purvalanol and roscovitine exerted different effects on cell survival and death mechanisms in LNCaP and DU145 cell due to their AR receptor status. Our studies show that co-treatment of rapamycin with CDK inhibitors inhibit prostate cancer cell viability more effectively than either agent alone, in part, by targeting the mTOR signaling cascade in AR (+) LNCaP cells. In this point, mTOR is a fine-tuning player in purvalanol and roscovitine-induced apoptosis and autophagy via regulation of PI3K/AKT and the downstream targets, which related with cell proliferation.


The coalescent and the genealogical process in geographically structured population.
We shall extend Kingman's coalescent to the geographically structured population model with migration among colonies. It is described by a continuous-time Markov chain, which is proved to be a dual process of the diffusion process of stepping-stone model. We shall derive a system of equations for the spatial distribution of a common ancestor of sampled genes from colonies and the mean time to getting to one common ancestor. These equations are solved in three particular models; a two-population model, the island model and the one-dimensional stepping-stone model with symmetric nearest-neighbour migration.


Are prior pregnancy outcomes relevant for models of fertility-specific distress or infertility helpseeking?
Women with prior pregnancy but no live birth are inconsistently termed as either 'primary infertile' or 'secondary infertile' in psychosocial studies of infertile women. The goal of this study was to discover whether infertile women who had experienced pregnancies but no live births were more similar in attitudes and behaviour to infertile women who had not experienced pregnancies or to those who had live births. We used the National Survey of Fertility Barriers (NSFB), which contains self-reported data from a probability-based sample of US women aged between 25 and 45, to accomplish our goal. In this cross-sectional analysis, infertile women who had not experienced pregnancies were compared on the basis of fertility-specific distress (FSD) and medical help-seeking for infertility to women who had had pregnancies with live births and women with pregnancies but no live births. Women were interviewed by telephone in their homes. Data of 1,027 women who had had an infertility episode within the past 10 years were analysed using multiple regression and logistic regression. Infertile women who had never been pregnant experience higher levels of FSD and were more likely to seek treatment than infertile women who had been pregnant, regardless of the outcome of the pregnancy.


Prediction of drug response in breast cancer using integrative experimental/computational modeling.
Nearly 30% of women with early-stage breast cancer develop recurrent disease attributed to resistance to systemic therapy. Prevailing models of chemotherapy failure describe three resistant phenotypes: cells with alterations in transmembrane drug transport, increased detoxification and repair pathways, and alterations leading to failure of apoptosis. Proliferative activity correlates with tumor sensitivity. Cell-cycle status, controlling proliferation, depends on local concentration of oxygen and nutrients. Although physiologic resistance due to diffusion gradients of these substances and drugs is a recognized phenomenon, it has been difficult to quantify its role with any accuracy that can be exploited clinically. We implement a mathematical model of tumor drug response that hypothesizes specific functional relationships linking tumor growth and regression to the underlying phenotype. The model incorporates the effects of local drug, oxygen, and nutrient concentrations within the three-dimensional tumor volume, and includes the experimentally observed resistant phenotypes of individual cells. We conclude that this integrative method, tightly coupling computational modeling with biological data, enhances the value of knowledge gained from current pharmacokinetic measurements, and, further, that such an approach could predict resistance based on specific tumor properties and thus improve treatment outcome.


Survival impact of integrative cancer care in advanced metastatic breast cancer.
Integrative cancer treatment is of substantial interest to many cancer patients. Research is needed to evaluate the effects of integrative treatment on patient outcomes. We report survival data for a consecutive case series of advanced metastatic breast cancer patients who received a comprehensive clinical program combining conventional treatments with nutrition and supplementation, fitness and mind-spirit instruction at the Block Center for Integrative Cancer Treatment. Treatment outcomes using integrative care for this disease have not previously been documented; survival data will thus contribute to decisions concerning future research directions and design. Ninety consecutive patients with metastatic breast cancer diagnosed during 1984-1997 who received chemotherapy at the integrative cancer center were included. Prognostic factors, treatments and survival from onset of metastases were determined from analysis of scans, labs, pathology and medical records. The log-rank test and Cox proportional hazards analyses were used, and a Kaplan-Meier curve was calculated. All patients had metastatic disease at baseline, 96% were relapsed and 52% had received prior chemotherapy for metastatic disease. Median age at onset of metastasis was 46 years. Median survival was 38 months (95% CI 27,48). Published literature on populations with somewhat more favorable prognostic factors treated in conventional clinics showed median survivals of 20 to 23 months. Through the 1990s, median survival reported in metastatic breast cancer trials or observations generally ranged from 12 to 24 months. Five-year survival was 27% for Center versus 17% for comparison patients. Despite a higher proportion of younger and relapsed patients, survival of metastatic breast cancer patients at the Center was approximately double that of comparison populations and possibly even higher compared to trials published during this period. Explanations for the advantage relative to conventional treatment alone may include the nutritional, nutraceutical, exercise and psychosocial interventions, individually or in combination; self-selection of patients cannot be ruled out. Further research to evaluate the impact of integrative breast cancer treatment on survival is warranted.


The RNA content of human sperm reflects prior events in spermatogenesis and potential post-fertilization effects.
Transcriptome analyses using high-throughput methodologies allow a deeper understanding of biological functions in different cell types/tissues. The present study provides an RNA-seq profiling of human sperm mRNAs and lncRNAs (messenger and long non-coding RNAs) in a well-characterized population of fertile individuals. Sperm RNA was extracted from twelve ejaculate samples under strict quality controls. Poly(A)-transcripts were sequenced and aligned to the human genome. mRNAs and lncRNAs were classified according to their mean expression values (FPKM: Fragments Per Kilobase of transcript per Million mapped reads) and integrity. Gene Ontology analysis of the Expressed and Highly Expressed mRNAs showed an involvement in diverse reproduction processes, while the Ubiquitously Expressed and Highly Stable mRNAs were mainly involved in spermatogenesis. Transcription factor enrichment analyses revealed that the Highly Expressed and Ubiquitously Expressed sperm mRNAs were primarily regulated by zinc-fingers and spermatogenesis-related proteins. Regarding the Expressed lncRNAs, only one-third of their potential targets corresponded to Expressed mRNAs and were enriched in cell-cycle regulation processes. The remaining two-thirds were absent in sperm and were enriched in embryogenesis-related processes. A significant amount of post-testicular sperm mRNAs and lncRNAs was also detected. Even though our study is solely directed to the poly-A fraction of sperm transcripts, results indicate that both sperm mRNAs and lncRNAs constitute a footprint of previous spermatogenesis events and are configured to affect the first stages of embryo development.


Modification of the optoelectronic properties of membranes via insertion of amphiphilic phenylenevinylene oligoelectrolytes.
We report on the modification of membranes by incorporation of phenylenevinylene oligoelectrolytes with the goal of tailoring their optical and electronic properties and their applications. A water-soluble distyrylstilbene oligoelectrolyte (DSSN+), capped at each end with nitrogen bound, terminally charged pendant groups, was synthesized. The photophysical and solvatochromatic properties of DSSN+ and the shorter distyrylbenzene analogue DSBN+ were probed and found to be useful for characterizing insertion into membranes based on phospholipid vesicle systems. A combination of UV/visible absorbance and photoluminescence spectroscopies, together with confocal microscopy, were employed to confirm membrane incorporation. Examination of the emission intensity profile in stationary multilamellar vesicles obtained with a polarized excitation source provides insight into the orientation of these chromophores within lipid bilayers and indicates that these molecules are highly ordered, such that the hydrophobic electronically delocalized region positions within the inner membrane with the long molecular axis perpendicular to the bilayer plane. Cyclic voltammetry experiments provide evidence that DSSN+ and DSBN+ facilitate transmembrane electron transport across lipid bilayers supported on glassy carbon electrodes. Additionally, the interaction with living microorganisms was probed. Fluorescence imaging indicates that DSSN+ and DSBN+ preferentially accumulate within cell membranes. Furthermore, notable increases in yeast microbial fuel cell performance were observed when employing DSSN+ as the electron transport mediator.


Impact of microcystin containing diets on physiological performance of Nile tilapia (Oreochromis niloticus) concerning stress and growth.
Diets containing Microcystis with considerable amounts of the cyanotoxin microcystin-LR (MC-LR) were fed to determine their impact on the physiological performance of the omnivorous Nile tilapia (Oreochromis niloticus) with regard to stress and growth performance. Four different diets were prepared based on a commercial diet (control, MC-5% [containing 5% dried Microcystis biomass], MC-20% [containing 20% dried Microcystis biomass], and Arthrospira-20% [containing 20% dried Arthrospira sp. biomass without toxin]) and fed to female Nile tilapia. Blood and tissue samples were taken after 1, 7, and 28 d, and MC-LR was quantified in gills, muscle, and liver by using high-performance liquid chromatography (HPLC). Only in the liver were moderate concentrations of MC-LR detected. The stress hormone cortisol and glucose were analyzed from plasma, suggesting that all modified diets caused only minor to moderate stress, which was confirmed by analyses of hepatic glycogen. In addition, the effects of the different diets on growth performance were investigated by determining gene expression of hypophyseal growth hormone (GH) and hepatic insulin-like growth factor-I (IGF-I). For all diets, quantitative reverse transcription-polymerase chain reaction (RT-qPCR) demonstrated no significant effect on gene expression of the major endocrine hormones of the growth axis, whereas classical growth data, including growth and feed conversion ratio, displayed slight inhibitory effects of all modified diets independent of their MC-LR content. However, no significant change was found in condition or hepatosomatic index among the various diets, so it seems feasible that dried cyanobacterial biomass might be even used as a component in fish diet for Nile tilapia, which requires further research in more detail.


USP33 regulates centrosome biogenesis via deubiquitination of the centriolar protein CP110.
Centrosome duplication is critical for cell division, and genome instability can result if duplication is not restricted to a single round per cell cycle. Centrosome duplication is controlled in part by CP110, a centriolar protein that positively regulates centriole duplication while restricting centriole elongation and ciliogenesis. Maintenance of normal CP110 levels is essential, as excessive CP110 drives centrosome over-duplication and suppresses ciliogenesis, whereas its depletion inhibits centriole amplification and leads to highly elongated centrioles and aberrant assembly of cilia in growing cells. CP110 levels are tightly controlled, partly through ubiquitination by the ubiquitin ligase complex SCF(cyclin F) during G2 and M phases of the cell cycle. Here, using human cells, we report a new mechanism for the regulation of centrosome duplication that requires USP33, a deubiquitinating enzyme that is able to regulate CP110 levels. USP33 interacts with CP110 and localizes to centrioles primarily in S and G2/M phases, the periods during which centrioles duplicate and elongate. USP33 potently and specifically deubiquitinates CP110, but not other cyclin-F substrates. USP33 activity antagonizes SCF(cyclin F)-mediated ubiquitination and promotes the generation of supernumerary centriolar foci, whereas ablation of USP33 destabilizes CP110 and thereby inhibits centrosome amplification and mitotic defects. To our knowledge, we have identified the first centriolar deubiquitinating enzyme whose expression regulates centrosome homeostasis by countering cyclin-F-mediated destruction of a key substrate. Our results point towards potential therapeutic strategies for inhibiting tumorigenesis associated with centrosome amplification.


Mutation Screening of Candidate Genes in Patients with Nonsyndromic Sagittal Craniosynostosis.
Craniosynostosis is a condition that includes the premature fusion of one or multiple cranial sutures. Among various craniosynostosis forms, sagittal nonsyndromic craniosynostosis is the most prevalent. Although different gene mutations have been identified in some craniosynostosis syndromes, the cause of sagittal nonsyndromic craniosynostosis remains largely unknown. To screen for candidate genes for sagittal nonsyndromic craniosynostosis, the authors sequenced DNA of 93 sagittal nonsyndromic craniosynostosis patients from a population-based study conducted in Iowa and New York states. FGFR1-3 mutational hotspots and the entire TWIST1, RAB23, and BMP2 coding regions were screened because of their known roles in human nonsyndromic or syndromic sagittal craniosynostosis, expression patterns, and/or animal model studies. The authors identified two rare variants in their cohort. A FGFR1 insertion c.730_731insG, which led to a premature stop codon, was predicted to abolish the entire immunoglobulin-like III domain, including the ligand-binding region. A c.439C>G variant was observed in TWIST1 at its highly conserved loop domain in another patient. The patient's mother harbored the same variant and was reported with jaw abnormalities. These two variants were not detected in 116 alleles from unaffected controls or seen in the several databases; however, TWIST1 variant was found in a low frequency of 0.000831 percent in Exome Aggregation Consortium database. The low mutation detection rate indicates that these genes account for only a small proportion of sagittal nonsyndromic craniosynostosis patients. The authors' results add to the perception that sagittal nonsyndromic craniosynostosis is a complex developmental defect with considerable genetic heterogeneity. Risk, II.


Lymphatic metastases with childhood rhabdomyosarcoma. A report from the Intergroup Rhabdomyosarcoma Study.
A review of 1415 patients without distant metastasis from the Intergroup Rhabdomyosarcoma Study (IRS) I and II revealed an overall 10% incidence of identified lymphatic spread at diagnosis, whereas 81 of 592 children with localized rhabdomyosarcoma who had grossly complete resection (and therefore with more complete pathologic data) had histologically proven lymphatic spread (14%). The percentage of patients in this latter group with nodal metastases was highest for the prostate (41%), paratesticular sites (26%), and genitourinary sites overall (24%). Sites with a small percentage of proven lymphatic involvement were the orbit (0%), nonorbital head and neck sites (7%), and truncal sites (3%), whereas the percentage of patients with nodal metastases from extremity lesions was 12%. The primary tumor mean diameter was significantly larger in the group with nodal metastases, but there was no evidence of a relationship between lymphatic spread and age, sex, or histologic subtype. Patients with lymph node metastases who had resection had a poorer survival rate (logrank P value = 0.001), with a 3-year survival estimate of 54%, compared with 78% for patients without lymphatic metastases. Patients with extremity lesions and positive lymph nodes also did poorly when compared with patients with normal nodes (P = 0.006), and a similar observation was made for patients with paratesticular sarcoma (P = 0.06).


A strategy to prevent signal losses, analyte decomposition, and fluctuating carbon contamination bands in surface-enhanced Raman spectroscopy.
Signal losses and fluctuating carbon contamination bands are "bottlenecks" in the application of surface-enhanced Raman spectroscopy (SERS) for reliable chemical analysis. They originate mainly from prolonged laser irradiation of the sample during data collection, which causes analyte decomposition and/or loss of the enhancing capabilities of the adsorption site. In this work, a laser illumination/signal collection technique, the "multiple points collection" (MPC) method is introduced to circumvent these problems. The MPC method is based on the use of a pair of galvanic mirrors to scan the laser beam rapidly and steadily across the sample surface. Each position is irradiated for <10 mus, at a rate of approximately 0.5 Hz. The SER spectrum is obtained by summing the signals collected from a large array of non-overlapping sample points. The MPC is compared with the conventional "single point collection" method, in which the laser beam is statically focused onto a particular spot and the scattered signals acquired. The MPC has the following advantages: (1) illumination and collection efficiencies are not compromised, (2) signal losses originating from analyte decomposition and/or alteration of the enhancing capabilities of the adsorption site are avoided, (3) high-quality SER spectra for analytes such as biomolecules and dipicolinic acid (a common marker for bacteria spores) can be easily obtained, and (4) the occurrence of broad amorphous carbon bands and the commonly observed temporal fluctuations in SERS are prevented. The success of the MPC is attributed to the reduction of local sample heating, as the time interval between the laser irradiations of a spot is much longer than the actual irradiation time itself.


Toward improved statistical methods for analyzing Cotinine-Biomarker health association data.
Serum cotinine, a metabolite of nicotine, is frequently used in research as a biomarker of recent tobacco smoke exposure. Historically, secondhand smoke (SHS) research uses suboptimal statistical methods due to censored serum cotinine values, meaning a measurement below the limit of detection (LOD). We compared commonly used methods for analyzing censored serum cotinine data using parametric and non-parametric techniques employing data from the 1999-2004 National Health and Nutrition Examination Surveys (NHANES). To illustrate the differences in associations obtained by various analytic methods, we compared parameter estimates for the association between cotinine and the inflammatory marker homocysteine using complete case analysis, single and multiple imputation, "reverse" Kaplan-Meier, and logistic regression models. Parameter estimates and statistical significance varied according to the statistical method used with censored serum cotinine values. Single imputation of censored values with either 0, LOD or LOD/√2 yielded similar estimates and significance; multiple imputation method yielded smaller estimates than the other methods and without statistical significance. Multiple regression modelling using the "reverse" Kaplan-Meier method yielded statistically significant estimates that were larger than those from parametric methods. Analyses of serum cotinine data with values below the LOD require special attention. "Reverse" Kaplan-Meier was the only method inherently able to deal with censored data with multiple LODs, and may be the most accurate since it avoids data manipulation needed for use with other commonly used statistical methods. Additional research is needed into the identification of optimal statistical methods for analysis of SHS biomarkers subject to a LOD.


Identifying conditions for elimination and epidemic potential of methicillin-resistant Staphylococcus aureus in nursing homes.
Residents of nursing homes are commonly colonized with methicillin-resistant Staphylococcus aureus (MRSA) but there is a limited understanding of the dynamics and determinants of spread in this setting. To address this gap, we sought to use mathematical modeling to assess the epidemic potential of MRSA in nursing homes and to determine conditions under which non-USA300 and USA300 MRSA could be eliminated or reduced in the facilities. Model parameters were estimated from data generated during a longitudinal study of MRSA in 6 Wisconsin nursing homes. The data included subject colonization status with strain-specific MRSA collected every 3 months for up to 1 year. Deterministic and stochastic co-colonization and single-strain models were developed to describe strain-specific dynamics of MRSA in these facilities. Basic reproduction numbers of strain-independent MRSA, non-USA300 and USA300 MRSA were estimated numerically. The impact of antibiotic use in the past 3 months on the prevalence of strain-specific MRSA and associated basic reproduction numbers were evaluated. Our models predicted that MRSA would persist in Wisconsin nursing homes, and non-USA300 would remain the dominant circulating strain. MRSA eradication was theoretically achievable by elimination of MRSA-positive admissions over the course of years. Substantial reductions in MRSA prevalence could be attained through marked increase in clearance rates or reduction in MRSA-positive admissions sustained over years. The basic reproduction number of strain-independent MRSA was 0.18 (95 % CI = 0.13-0.23). Recent antibiotic use increased the prevalence of strain-specific MRSA and associated basic reproduction numbers, but was unlikely to lead to an outbreak. Based on our model, MRSA elimination from nursing homes, while theoretically possible, was unlikely to be achieved in practice. Decolonization therapy that can sustain higher clearance rates or lower MRSA-positive introductions over years may reduce strain-specific prevalence of MRSA in the facilities, and antibiotic stewardship may contribute to this effort. Large-scale MRSA outbreaks were unlikely in this setting.


Expression of bcl-2 in bladder neoplasms is a cell lineage associated and p53-independent event.
To investigate bcl-2 and p53 protein expression in hyperplastic, metaplastic and neoplastic epithelia of the urinary bladder in relation to cell lineages (transitional versus glandular epithelia). Formalin fixed, paraffin wax embedded archival tissue blocks of 29 transitional cell carcinomas (TCC), 11 adenocarcinomas, five specimens of cystitis glandularis, four papillomas, and seven samples of morphologically normal bladder mucosa were examined immunohistochemically with antibodies specific to bcl-2 and p53. Consecutive sections were used to assess co-expression of the two proteins. bcl-2 protein was expressed heterogeneously in basal cells of the normal transitional epithelium, whereas p53 was rarely detectable in either normal or hyperplastic epithelium. Of the 29 TCCs, 20 (69%) expressed immunodetectable p53 which was positively associated with grade. In contrast, bcl-2 was detected in four (14%) TCCs and its expression was not associated with grade. bcl-2 was expressed constitutively in all five specimens of cystitis glandularis and in all adenocarcinomas; p53 was co-expressed in most of the latter. There was no association between bcl-2 and p53 protein expression in the TCCs. Expression of bcl-2 protein correlated negatively with grade of adenocarcinoma. In bladder adenocarcinomas, bcl-2 expression correlated negatively with tumour grade whereas p53 was associated positively with tumour grade. The association of bcl-2 with cystitis glandularis and adenocarcinoma but not TCC suggests that it may be involved in triggering a lineage switch converting transitional epithelium to a glandular phenotype.


Sub-proteome differential display: single gel comparison by 2D electrophoresis and mass spectrometry.
Two-dimensional (2D) gel electrophoresis and mass spectrometry (MS) have been used in comparative proteomics but inherent problems of the 2D electrophoresis technique lead to difficulties when comparing two samples. We describe a method (sub-proteome differential display) for comparing the proteins from two sources simultaneously. Proteins from one source are mixed with radiolabelled proteins from a second source in a ratio of 100:1. These combined proteomes are fractionated simultaneously using column chromatographic methods, followed by analysis of the pre-fractionated proteomes (designated sub-proteomes) using 2D gel electrophoresis. Silver staining and (35)S autoradiography of a single gel allows precise discrimination between members of each sub-proteome, using commonly available computer software. This is followed by MS identification of individual proteins. We have demonstrated the utility of the technology by identifying the product of a transfected gene and several proteins expressed differentially between two renal carcinoma proteomes. The procedure has the capacity to enrich proteins prior to 2D electrophoresis and provides a simple, inexpensive approach to compare proteomes. The single gel approach eliminates differences that might arise if separate proteome fractionations or 2D gels are employed.


Underwater linear polarization: physical limitations to biological functions.
Polarization sensitivity is documented in a range of marine animals. The variety of tasks for which animals can use this sensitivity, and the range over which they do so, are confined by the visual systems of these animals and by the propagation of the polarization information in the aquatic environment. We examine the environmental physical constraints in an attempt to reveal the depth, range and other limitations to the use of polarization sensitivity by marine animals. In clear oceanic waters, navigation that is based on the polarization pattern of the sky appears to be limited to shallow waters, while solar-based navigation is possible down to 200-400 m. When combined with intensity difference, polarization sensitivity allows an increase in target detection range by 70-80% with an upper limit of 15 m for large-eyed animals. This distance will be significantly smaller for small animals, such as plankton, and in turbid waters. Polarization-contrast detection, which is relevant to object detection and communication, is strongly affected by water conditions and in clear waters its range limit may reach 15 m as well. We show that polarization sensitivity may also serve for target distance estimation, when examining point source bioluminescent objects in the photic mesopelagic depth range.


A temperature gradient method for lipid phase diagram construction using time-resolved x-ray diffraction.
A method that enables temperature-composition phase diagram construction at unprecedented rates is described and evaluated. The method involves establishing a known temperature gradient along the length of a metal rod. Samples of different compositions contained in long, thin-walled capillaries are positioned lengthwise on the rod and "equilibrated" such that the temperature gradient is communicated into the sample. The sample is then moved through a focused, monochromatic synchroton-derived x-ray beam and the image-intensified diffraction pattern from the sample is recorded on videotape continuously in live-time as a function of position and, thus, temperature. The temperature at which the diffraction pattern changes corresponds to a phase boundary, and the phase(s) existing (coexisting) on either side of the boundary can be identified on the basis of the diffraction pattern. Repeating the measurement on samples covering the entire composition range completes the phase diagram. These additional samples can be conveniently placed at different locations around the perimeter of the cylindrical rod and rotated into position for diffraction measurement. Temperature-composition phase diagrams for the fully hydrated binary mixtures, dimyristoylphosphatidylcholine (DMPC)/dipalmitoylphosphatidylcholine (DPPC) and dipalmitoylphosphatidylethanolamine (DPPE)/DPPC, have been constructed using the new temperature gradient method. They agree well with and extend the results obtained by other techniques. In the DPPE/DPPC system structural parameters as a function of temperature in the various phases including the subgel phase are reported. The potential limitations of this steady-state method are discussed.


Multicenter assessment of surgical outcomes in adult spinal deformity patients with severe global coronal malalignment: determination of target coronal realignment threshold.
The impact of global coronal malalignment (GCM; C7 plumb line-midsacral offset) on adult spinal deformity (ASD) treatment outcomes is unclear. Here, the authors' primary objective was to assess surgical outcomes and complications in patients with severe GCM, with a secondary aim of investigating potential surgical target coronal thresholds for optimal outcomes. This is a retrospective analysis of a prospective multicenter database. Operative patients with severe GCM (≥ 1 SD above the mean) and a minimum 2-year follow-up were identified. Demographic, surgical, radiographic, health-related quality of life (HRQOL), and complications data were analyzed. Of 691 potentially eligible operative patients (mean GCM 4 ± 3 cm), 80 met the criteria for severe GCM ≥ 7 cm. Of these, 62 (78%; mean age 63.7 ± 10.7 years, 81% women) had a minimum 2-year follow-up (mean follow-up 3.3 ± 1.1 years). The mean ASD-Frailty Index was 3.9 ± 1.5 (frail), 50% had undergone prior fusion, and 81% had concurrent severe sagittal spinopelvic deformity with GCM and C7-S1 sagittal vertical axis (SVA) positively correlated (r = 0.313, p = 0.015). Surgical characteristics included posterior-only (58%) versus anterior-posterior (42%) approach, mean fusion of 13.2 ± 3.8 levels, iliac fixation (90%), 3-column osteotomy (36%), operative duration of 8.3 ± 3.0 hours, and estimated blood loss of 2.3 ± 1.7 L. Final alignment and HRQOL significantly improved (p < 0.01): GCM, 11 to 4 cm; maximum coronal Cobb angle, 43° to 20°; SVA, 13 to 4 cm; pelvic tilt, 29° to 23°; pelvic incidence-lumbar lordosis mismatch, 31° to 5°; Oswestry Disability Index, 51 to 37; physical component summary of SF-36 (PCS), 29 to 37; 22-Item Scoliosis Research Society Patient Questionnaire (SRS-22r) Total, 2.6 to 3.5; and numeric rating scale score for back and leg pain, 7 to 4 and 5 to 3, respectively. Residual GCM ≥ 3 cm was associated with worse SRS-22r Appearance (p = 0.04) and SRS-22r Satisfaction (p = 0.02). The minimal clinically important difference and/or substantial clinical benefit (MCID/SCB) was met in 43%-83% (highest for SRS-22r Appearance [MCID 83%] and PCS [SCB 53%]). The severity of baseline GCM (≥ 2 SD above the mean) significantly impacted postoperative SRS-22r Satisfaction and MCID/SCB improvement for PCS. No significant partial correlations were demonstrated between GCM or SVA correction and HRQOL improvement. There were 89 total complications (34 minor and 55 major), 45 (73%) patients with ≥ 1 complication (most commonly rod fracture [19%] and proximal junctional kyphosis [PJK; 18%]), and 34 reoperations in 22 (35%) patients (most commonly for rod fracture and PJK). Study results demonstrated that ASD surgery in patients with substantial GCM was associated with significant radiographic and HRQOL improvement despite high complication rates. MCID improvement was highest for SRS-22r Appearance/Self-Image. A residual GCM ≥ 3 cm was associated with a worse outcome, suggesting a potential coronal realignment target threshold to assist surgical planning.


Hydrogen supplemented air inhalation reduces changes of prooxidant enzyme and gap junction protein levels after transient global cerebral ischemia in the rat hippocampus.
Transient global cerebral ischemia (TGCI) occurs during acute severe hypotension depriving the brain of oxygen and glucose for a short period of time. During reperfusion, several mechanisms can induce secondary neuronal damage, including the increased production of reactive oxygen species (ROS). Hydrogen gas-enriched air inhalation is a neuroprotective approach with proven antioxidant potential, which has not yet been examined in TGCI. Accordingly, we set out to describe the effect of inhalation of 2.1% hydrogen supplemented room air (H(2)-RA) in comparison with a well studied neuroprotective agent, rosiglitazone (RSG) in a TGCI rat model. Male Wistar rats were exposed to TGCI (n=26) or sham operation (n=26), while a third group served as intact control (naive, n=5). The operated groups were further divided into non-treated, H(2)-RA, RSG (6 mg/kg i.v.) and vehicle treated animals. Tissue samples from the hippocampus and frontal cortex were taken 3 days following surgery. Western blot analysis was applied to determine the expressions of cyclooxygenase-2 (COX-2), neuronal and endothelial nitric oxide synthase (nNOS and eNOS, respectively), manganese superoxide dismutase (MnSOD) and glial connexin proteins: connexin 30 and connexin 43. The expressions of COX-2, and connexin proteins were upregulated, while nNOS was downregulated 3 days after TGCI. Both RSG and H(2)-RA prevented the changes of enzyme and connexin levels. Considering the lack of harmful side effects, inhalation of H(2)-RA can be a promising approach to reduce neuronal damage after TGCI.


Overview of the use of the U.S. EPA exposure factors handbook.
Risk assessments are important components of the decision making process. At hazardous waste sites, they are used as tools to determine appropriate cleanup levels. Therefore, it is critical that the best up-to-date methods, models, and exposure data are available to the exposure and risk assessor to realistically estimate the potential for human and ecological exposures to environmental contaminants. The EPA Exposure Factors Handbook published in 1997 is a tool available to exposure assessors which summarizes statistical data on exposure factors necessary to conduct human health exposure assessments. Since it was first published by EPA in 1989, the handbook has been the primary source of data for human exposure assessments. The purpose of this paper is to provide an overview of the handbook, its impact, applications, discussion about data gaps, and future directions.


A multicenter evaluation of a compact, sterile, single-use pressure transducer for central venous catheter placement.
Inadvertent arterial placement of a large-bore catheter during attempted placement of a central venous catheter (CVC) occurs at a rate of 0.1% to 1.0% and may result in hemorrhage, pseudoaneurysm, stroke, or death. Ultrasound guidance or observation of color and pulsatility of blood are not reliable methods for avoiding this serious complication. Measurement of pressure in the needle or short plastic catheter before insertion of the guidewire has been shown to be highly reliable; however, traditional pressure measurement methodology is cumbersome. Recently a compact, sterile, single-use pressure transducer with an integrated digital display has become available. In this study, we evaluated the performance of this new device (Compass® Vascular Access). In this prospective, observational study at 4 academic medical centers 298 CVCs were placed. Pressure was measured using the Compass transducer before and after guidewire insertion. Other details of the procedure were at the discretion of the clinician. Data describing the CVC placement and any complications were collected. Trainees placed 279 of 298 CVCs. Ultrasound guidance was used for 286 of 298 CVCs. Seven of the CVC placements occurred in the intensive care unit, with the balance occurring in the operating room. Ten of the CVCs were placed in a subclavian vein, with the balance being internal jugular vein. Two hundred seventy-four of 298 CVCs were placed on the right side. Venous pressure measured before and after guidewire insertion was 7.2 ± 4.3 (SD) and 6.5 ± 4.3 (SD) mm Hg respectively (P = 0.03). The satisfaction score recorded by the physician performing the procedure was 8.0 ± 2.1 (SD; visual analog scale 1-10, 10 being most satisfying). There were 5 inadvertent arterial punctures (1.7%). Ultrasound guidance was used in all 5 cases of arterial puncture. All of the arterial punctures were recognized before guidewire insertion by measurement of arterial pressure with the Compass transducer. No guidewires or CVC catheters were placed in arteries. The Compass pressure transducer for CVC placement performed as intended in 298 cases from 4 academic medical centers. There were 5 inadvertent arterial punctures despite the use of ultrasound guidance, all of which were correctly identified by pressure measurement using the Compass. The device was easily used by trainees, and users expressed a positive level of satisfaction.


Increased Circulating T Follicular Helper Cells Are Inhibited by Rituximab in Neuromyelitis Optica Spectrum Disorder.
Neuromyelitis optica spectrum disorder (NMOSD) is a severe autoimmune disease of the central nervous system. The existence of autoantibody targeting aquaporin-4 (AQP4-Ab) indicates the involvement of humoral immunity in the pathogenesis of this disease. Rituximab (RTX), a monoclonal antibody against CD20, has been used to treat NMOSD by depleting circulating B cells and overall satisfactory outcome has been achieved. Although T follicular helper cells have been proved to regulate B cell activation and antibody production, the role of these cells in NMOSD and the impact of RTX treatment on these cells remain less understood. In this study, we found that frequencies of circulating T follicular helper (cTfh) cells and B cells together with the related cytokines, IL-21 and IL-6, were closely correlated with disease activity of NMOSD. Furthermore, B cell depletion with RTX treatment inhibited the expansion of cTfh cells, and these effects were achieved through eliminating IL-6-producing B cells and blocking the direct contact between cTfh cells and B cells. These findings imply the complicated cross talk between cTfh cells and B cells and may provide a novel therapeutic target for NMOSD.


Integrative genomics analysis of genes with biallelic loss and its relation to the expression of mRNA and micro-RNA in esophageal squamous cell carcinoma.
Genomic instability plays an important role in human cancers. We previously characterized genomic instability in esophageal squamous cell carcinomas (ESCC) in terms of loss of heterozygosity (LOH) and copy number (CN) changes in tumors. In the current study we focus on biallelic loss and its relation to expression of mRNA and miRNA in ESCC using results from 500 K SNP, mRNA, and miRNA arrays in 30 cases from a high-risk region of China. (i) Biallelic loss was uncommon but when it occurred it exhibited a consistent pattern: only 77 genes (<0.5%) showed biallelic loss in at least 10% of ESCC samples, but nearly all of these genes were concentrated on just four chromosomal arms (i.e., 42 genes on 3p, 14 genes on 9p, 10 genes on 5q, and seven genes on 4p). (ii) Biallelic loss was associated with lower mRNA expression: 52 of the 77 genes also had RNA expression data, and 41 (79%) showed lower expression levels in cases with biallelic loss compared to those without. (iii) The relation of biallelic loss to miRNA expression was less clear but appeared to favor higher miRNA levels: of 60 miRNA-target gene pairs, 34 pairs (57%) had higher miRNA expression with biallelic loss than without, while 26 pairs (43%) had lower miRNA expression. (iv) Finally, the effect of biallelic loss on the relation between miRNA and mRNA expression was complex. Biallelic loss was most commonly associated with a pattern of elevated miRNA and reduced mRNA (43%), but a pattern of both reduced miRNA and mRNA was also common (35%). Our results indicate that biallelic loss in ESCC is uncommon, but when it occurs it is localized to a few specific chromosome regions and is associated with reduced mRNA expression of affected genes. The effect of biallelic loss on miRNA expression and on the relation between miRNA and mRNA expressions was complex.


Temporal Modulation of the Response of Sensory Fibers to Paired-Pulse Stimulation.
Multi-channel nerve cuff electrode arrays can provide sensory feedback to prosthesis users. To develop efficacious stimulation protocols, an understanding of the impact that spatio-temporal patterned stimulation can have on the response of sensory fibers is crucial. We used experimental and modelling methods to investigate the response of nerve fibers to paired-pulse stimulation. Nerve cuff electrode arrays were implanted for stimulation of the sciatic nerves of rats and the sensory compound action potentials were recorded from the L4 dorsal root. A model of the nerve cuff electrode array and sciatic nerve was also developed. The experimental and modelling results were compared. Experiments showed that it took 8 ms for the sensory fibers to completely recover from a conditioning stimulus, regardless of the relative position of the electrodes used for stimulation. The results demonstrate that the electrodes on the cuff cannot be considered independent. Additionally, at 120% of the threshold, there is a large overlap in the fibers that were activated by the different electrodes. If a stimulus paradigm considered the electrodes as independent, stimuli from the different electrodes would need to be interleaved, and the intervals between the stimuli should be greater than 8 ms.


Intranasal murine model of Bordetella pertussis infection. I. Prediction of protection in human infants by acellular vaccines.
Bicomponent, tricomponent and pertactin DTPa vaccines were tested in sublethal aerosol, and lethal and sublethal intranasal murine Bordetella pertussis respiratory challenge models. Pertactin and bicomponent vaccines induced protective immunity against lethality but with little or no bacterial clearance. Intranasal challenge discriminated in a reproducible, statistically significant manner between the efficacies of bicomponent and tricomponent DTPa, in agreement with clinical trial data. This discrimination was not observed in the aerosol challenge. Pertactin had a synergistic effect with bicomponent DTPa. Intranasal challenge may be useful as part of the preclinical evaluation of new acellular pertussis formulations or DTPa-based combinations.


Human endotoxin tolerance is associated with enrichment of the CD14+ CD16+ monocyte subset.
Prior exposure to lipopolysaccharides (LPS) induces a state of cell resistance to subsequent LPS restimulation, known as endotoxin tolerance, mainly by repressing the expression of pro-inflammatory cytokines. We established an endotoxin tolerance model in human monocytes Endotoxin-tolerant cells showed a decrease in IκBα degradation and diminished expression of Tumor necrosis factor (TNF) (both messenger RNA [mRNA] and protein content). The myeloid differentiation factor 88 (MyD88)/MyD88 splice variant (MyD88s) ratio, an indirect way to test the Toll-like receptor 4 (TLR4) MyD88-dependent signaling cascade, did not change in endotoxin-tolerant cells when compared to LPS-stimulated or -unstimulated ones. Remarkably, cell population analysis indicated a significant increase of the CD14+ CD16+ subset only under the endotoxin-tolerant condition. Furthermore, endotoxin-tolerant cells produced higher amounts of C-X-C motif chemokine 10 (CXCL10), a typical MyD88-independent cytokine.


Hormone therapy, gender affirmation surgery, and their association with recent suicidal ideation and depression symptoms in transgender veterans.
Access to transition-related medical interventions (TRMIs) for transgender veterans has been the subject of substantial public interest and debate. To better inform these important conversations, the current study investigated whether undergoing hormone or surgical transition intervention(s) relates to the frequency of recent suicidal ideation (SI) and symptoms of depression in transgender veterans. This study included a cross-sectional, national sample of 206 self-identified transgender veterans. They self-reported basic demographics, TRMI history, recent SI, and symptoms of depression through an online survey. Significantly lower levels of SI experienced in the past year and 2-weeks were seen in veterans with a history of both hormone intervention and surgery on both the chest and genitals in comparison with those who endorsed a history of no medical intervention, history of hormone therapy but no surgical intervention, and those with a history of hormone therapy and surgery on either (but not both) the chest or genitals when controlling for sample demographics (e.g., gender identity and annual income). Indirect effect analyses indicated that lower depressive symptoms experienced in the last 2-weeks mediated the relationship between the history of surgery on both chest and genitals and SI in the last 2-weeks. Results indicate the potential protective effect that TRMI may have on symptoms of depression and SI in transgender veterans, particularly when both genitals and chest are affirmed with one's gender identity. Implications for policymakers, providers, and researchers are discussed.


Phonological encoding is not contingent on semantic feature retrieval: an electrophysiological study on object naming.
In the present study, the authors examined with event-related brain potentials whether phonological encoding in picture naming is mediated by basic semantic feature retrieval or proceeds independently. In a manual 2-choice go/no-go task the choice response depended on a semantic classification (animal vs. object) and the execution decision was contingent on a classification of name phonology (vowel vs. consonant). The introduction of a semantic task mixing procedure allowed for selectively manipulating the speed of semantic feature retrieval. Serial and parallel models were tested on the basis of their differential predictions for the effect of this manipulation on the lateralized readiness potential and N200 component. The findings indicate that phonological code retrieval is not strictly contingent on prior basic semantic feature processing.


The three official language versions of the Declaration of Helsinki: what's lost in translation?
The Declaration of Helsinki, the World Medical Association's (WMA's) statement of ethical guidelines regarding medical research, is published in the three official languages of the WMA: English, French and Spanish. A detailed comparison of the three official language versions was carried out to determine ways in which they differed and ways in which the wording of the three versions might illuminate the interpretation of the document. There were many minor linguistic differences between the three versions. However, in paragraphs 1, 6, 29, 30 and in the note of clarification to paragraph 29, there were differences that could be considered potentially significant in their ethical relevance. Given the global status of the Declaration of Helsinki and the fact that it is translated from its official versions into many other languages for application to the ethical conduct of research, the differences identified are of concern. It would be best if such differences could be eliminated but, at the very least, a commentary to explain any differences that are unavoidable on the basis of language or culture should accompany the Declaration of Helsinki. This evidence further strengthens the case for international surveillance of medical research ethics as has been proposed by the WMA.


Tension measurement of artificial chordae tendinae implanted between the anterior mitral valve leaflet and the left ventricular apex: an in vitro study.
: A newly developed surgical technique allows implantation of artificial chordae tendinae between a mitral valve leaflet and the left ventricular (LV) apex in a closed-chest, off-pump mitral valve repair operation. However, this orientation causes tension vectors on the mitral valve leaflet in a different direction than those of native chordae. The purpose of this study was to investigate the relationship between systolic LV pressure and the tension of artificial chordae secured at the ventricular apex to assess if the altered tension vectors on the mitral leaflet resulted in excessive stress on the chordae/leaflet system. : Freshly harvested porcine mitral valves were suspended in a mechanical LV simulator. Two 5-0 Gore-Tex sutures were secured to the free edge of the middle scallop of the anterior leaflet adjacent to the insertion point of two major native chordae. The native chordae were then cut. A simulator pump was activated at different pressure levels, and the tension of the artificial chordae was measured with a digital tensiometer during several cardiac cycles. : The numerical relationship between chordal tension (in Newtons, N) and LV pressure (in mm Hg/100) averaged at 0.8 (range 0.6-1.05). This tension/pressure relationship was similar in order of magnitude and range to that measured in native chordae in previously published studies. : In this in vitro model, orienting artificial chordae in a direction mimicking implantation between the valve and the LV apex does not result in excessive tension stress on either the chordae or the mitral leaflet.


Working memory mediates the relationship between intellectual enrichment and long-term memory in multiple sclerosis: an exploratory analysis of cognitive reserve.
Some individuals with multiple sclerosis (MS) show decrements in long-term memory (LTM) while other individuals do not. The theory of cognitive reserve suggests that individuals with greater pre-morbid intellectual enrichment are protected from disease-related cognitive decline. How intellectual enrichment affords this benefit remains poorly understood. The present study tested an exploratory meditational hypothesis whereby working memory (WM) capacity may mediate the relationship between intellectual enrichment and verbal LTM decline in MS. Intellectual enrichment, verbal LTM, and WM capacity were estimated with the Wechsler Test of Adult Reading and Peabody Picture Vocabulary Test, delayed recall of the Hopkins Verbal Learning Test-Revised and Logical Memory of the Wechsler Memory Scale, and Digit Span Total, respectively. Intellectual enrichment predicted LTM (B=.54; p=.003) and predicted WM capacity (B=.91; p<.001). WM capacity predicted LTM, (B=.44; p<.001) and fully mediated the relationship between intellectual enrichment (B=.24; p=.27) and LTM (B=.33, p=.03), Sobel test, Z=3.31, p<.001. These findings implicate WM capacity as an underlying mechanism of cognitive reserve and are an initial first step in understanding the relationship between intellectual enrichment, WM, and LTM in MS.