Use of emergency department services for hand, wrist, and forearm fractures in the United States in 2008.
The objective of this study was to determine demographic characteristics and epidemiology of hand, wrist, and forearm fracture patients treated in the emergency departments (EDs) to identify the at-risk populations. The Nationwide Emergency Department Sample database collected by the Agency for Healthcare Research and Quality was used to estimate ED visits for hand, wrist, and forearm fractures during 2008. The overall rate of ED visits for these injuries was 54 per 10,000 population. Children aged 6 to 15 had the highest rate of ED visits at 124 per 10,000. A secondary increase in ED visits occurred at age 50 and rose with age. Metacarpals were the most common fracture location in adults aged 16 to 25, while radius and ulna were the most common fracture location in all other age groups. Demographic analysis identified children, adult males aged 16 to 25, and the elderly as target populations for preventive interventions.


Identification of sulfhydryl-containing proteins and further evaluation of the selenium-tagged redox homeostasis-regulating proteins.
Chemoproteomic profiling of sulfhydryl-containing proteins has consistently been an attractive research hotspot. However, there remains a dearth of probes that are specifically designed for sulfhydryl-containing proteins, possessing sufficient reactivity, specificity, distinctive isotopic signature, as well as efficient labeling and evaluation capabilities for proteins implicated in the regulation of redox homeostasis. Here, the specific selenium-containing probes (Se-probes) in this work displayed high specificity and reactivity toward cysteine thiols on small molecules, peptides and purified proteins and showed very good competitive effect of proteins labeling in gel-ABPP. We identified more than 6000 candidate proteins. In TOP-ABPP, we investigated the peptide labeled by Se-probes, which revealed a distinct isotopic envelope pattern of selenium in both the primary and secondary mass spectra. This unique pattern can provide compelling evidence for identifying redox regulatory proteins and other target peptides. Furthermore, our examiation of post-translational modification (PTMs) of the cysteine site residues showed that oxidation PTMs was predominantly observed. We anticipate that Se-probes will enable broader and deeper proteome-wide profiling of sulfhydryl-containing proteins, provide an ideal tool for focusing on proteins that regulate redox homeostasis and advance the development of innovative selenium-based pharmaceuticals.


Gastroresistant oral peptide for fluorescence imaging of colonic inflammation.
The use of molecular markers for inflammation in the gastrointestinal tract could empower optical imaging modalities for early diagnosis and eventually personalized timely treatments. A major hurdle to the widespread use of functional fluorescence imaging is the absence of suitable contrast agents, in particular to be administered via the oral route due to the usual proteolytic susceptibility of the biomarkers. By designing a retro-inverso peptide, starting from a previously described sequence specific for N-cadherin, we achieved resistance to gastrointestinal degradation and even slightly improved specificity towards the target, both in ex vivo and in vivo experimental colitis. Simulations at fundamental molecular level suggested that the introduced retro-inverso modifications did not affect the folding of the peptide, leaving its ability to interact with the binding pocket of the monomeric N-cadherin unaltered, even when fluorescently labeled. Possible further derivatization of this sequence could be envisaged to further extend the potential of the designed retro-inverso peptide as diagnostic or theranostic agent for the oral route.


Advances in local and ablative treatment of oligometastasis in prostate cancer.
Oligometastasis is a state of limited metastatic disease that may be amenable to aggressive local therapy to achieve long-term survival. This review aims to explore the role of ablative radiotherapy and surgical management of prostate cancer (CaP) patients with oligometastasis. We performed a systematic review of the literature from November 2003 to November 2013 in the PubMed and EMBASE databases using structured search terms. From our literature search, we identified 13 cases of oligometastatic CaP managed by surgery. The longest disease-free survival documented was 12 years following pulmonary metastasectomy. We also found 12 studies using radiotherapy to treat oligometastatic CaP with median follow-up ranging from 6 to 43 months. Local control rates and overall survival at 3 years range from 66 to 90% and from 54 to 92%, respectively. Most patients did not report any significant toxicity. The limited current literature suggests oligometastatic CaP may be amenable to more aggressive local ablative therapy to achieve prolonged local control and delay to androgen deprivation therapy (ADT). There is a larger body of evidence supporting the use of radiotherapy than surgery in this disease state. However, no direct comparison with ADT is available to suggest an improvement in overall survival. Further studies are required to determine the role of aggressive-targeted local therapy in oligometastatic CaP.


[Modified normothermic avascular hepatic resection].
The conventional Heaney's avascular hepatic resection was modified based on an experimental study on normothermic total hepatic vascular exclusion in dog. The modification was cancellation of simultaneous exclusion of the abdominal aorta and proper prolongation of total hepatic vascular exclusion time. The method was successfully used in 3 patients, 2 with primary hepatocellular carcinoma involving the second porta hepatis and 1 with a huge hepatic cavernous hemangioma in segments VII and VIII. The clinical application of this method is presented and discussed. The modified Heaney's technique, being simpler and safer than the conventional one, may increase the resection rate for central hepatic tumors (segments I, IV, V and VIII according to Couinaud's classification).


Deciphering indented impressions on plastic.
The questioned document laboratory is often called upon to decipher writing that has been erased, obliterated, or that has faded. In cases like these, the original writing is no longer legible to the naked eye, but may be enhanced using various light sources. Certain remnants of the ink's components absorb into the substrate's fibers and can be visualized, usually as luminescence or absorbance. A case is described here that involved the theft of a credit card. An empty plastic credit card holder was found in the possession of a suspect, and as submitted for examination. Indented impressions could be discerned on its clear plastic window and presumably originated from the credit card that had been held in the envelope. These indented impressions were deciphered in the hope that they would reveal enough details from the credit card to establish a connection between the plastic envelope and the stolen credit card. With methods generally utilized in the toolmarks and materials laboratory and the photography laboratory of the Israel Police, most of the indented impressions on the plastic were deciphered and a connection between the plastic envelope and the stolen credit card was demonstrated.


Functional opsonic activity of human serum antibodies to inner core lipopolysaccharide (galE) of serogroup B meningococci measured by flow cytometry.
A recently described flow cytometric opsonophagocytic assay (OPA) was adapted to quantify the functional activity of serum antibodies specifically directed against serogroup B inner core lipopolysaccharide (LPS) of Neisseria meningitidis. The percentage of human peripheral polymorphonuclear leukocytes and monocytes (PMNms) ingesting fluorescently labeled, ethanol-fixed N. meningitidis organisms (phagocytic activity) in the presence of human sera was measured to reflect the serum opsonic activity against the bacterium. The contribution to opsonophagocytic activity of antibodies to inner core LPS was estimated by comparing the opsonic activities of adult and infant sera before and after adsorbing anti-LPS antibodies from the sera using purified LPS extracted from an LPS mutant (galE) of N. meningitidis strain MC58 (B:15:P1.7,16:L3). The specificity of the assay was further investigated using monoclonal antibody (MAb) B5, which binds to an inner core LPS epitope of N. meningitidis. A dose-dependent decrease in phagocytic activity was observed when MAb B5 was incubated with LPS from an inner core LPS (galE) mutant. Similarly, the number of PMNms ingesting fluorescently labeled polystyrene beads coated with inner core (galE) LPS decreased in a dose-dependent fashion when MAb B5 was incubated with various concentrations of the homologous inner core LPS. Strong correlations were found between the concentration of serum antibodies to inner core LPS (galE) versus the phagocytic activity using healthy adult sera (r(2) = 0.89). There was a correlation between phagocytic ingestion and initiation of intracellular oxidative burst (r(2) = 0.99) using polystyrene beads coated with inner core LPS and opsonized with the same sera using the oxidative burst indicator system dihydrorhodamine123/rhodamine 123. OPA results were also found to correlate closely with the results of the serum bactericidal assay using MAb B5 against the N. meningitidis MC58 galE mutant in the presence of human complement (r(2) = 0.994, P = 0.003, two-tailed test). These studies demonstrate that functional antibodies are produced in humans against meningococcal inner core LPS and that the OPA is a useful approach to study the opsonic activity of antibodies to inner core LPS in health and disease.


EGF‑stimulated AKT activation is mediated by EGFR recycling via an early endocytic pathway in a gefitinib‑resistant human lung cancer cell line.
The receptor tyrosine kinase epidermal growth factor receptor (EGFR) and its ligand epidermal growth factor (EGF) are known to play important roles in malignant tumor cells, and the EGFR signaling pathway is one of the most important targets in various tumors, including non-small cell lung cancer (NSCLC). We reported recently that an aberration in certain steps of EGF-stimulated phosphorylated epidermal growth factor receptor (pEGFR) endocytic trafficking from the early endosomes to the late endosomes occurs in the gefitinib-resistant NSCLC cells, in which large amounts of sorting nexin 1 (SNX1) are colocalized with EGFR in the aggregated early endosomes where the internalized pEGFR is also accumulated of these cells. To further investigate the role of SNX1 in EGF‑stimulated pEGFR endocytosis, followed by downstream signaling leading to the activation of phosphatidylinositol 3-kinase (PI3K)--the serine/threonine kinase AKT pathway, we examined the effect of depletion of SNX1 knock-down expression by siRNA and an inhibition of targeting membrane recycling using monensin. Using immunofluorescence, we observed an efficient endocytic transport of pEGFR from early endosomes to late endosomes/lysosomes after EGF-stimulation in the cells transfected with siRNA‑SNX1, whereas the delayed endocytic delivery of pEGFR was evident in the siRNA-control-transfected cells. Furthermore, a large amount of endocytosed pEGFR was accumulated in the presence of monensin in the early endosomes of the SNX1 knock-down cells. In western blot analysis, EGF stimulation of both control and cells transfected with siRNA-SNX1 resulted in rapid phosphorylation of EGFR and enhanced AKT phosphorylation. Monensin-dependent inhibition of AKT phosphorylation was stronger in SNX1 knock-down cells than in controls. In contrast, however, monensin had no effect on AKT phosphorylation triggered by activation of the MET receptor tyrosine kinase. Collectively, we suggest that EGF-stimulated recycling of EGFR to the plasma membrane induces downstream signaling leading to AKT phosphorylation. Suppression of EGFR membrane recycling by SNX1 appears to be critical for the activation of EGFR/PI3K/AKT signaling pathway in human lung cancer cells.


Summary of ongoing clinical trials of platelet-active drugs in cardiovascular disease.
Design features of 17 ongoing clinical trials testing various platelet-active drug regimens in the primary or secondary prevention of coronary or cerebrovascular disease are summarized. The likelihood of detecting biologically important effects given their presence varies widely among the trial; a few have very modest statistical power; others are more robust. The majority of the trials will conclude follow-up and report results within the next several years.


[Sentinel lymph nodes in colorectal cancer].
Metastasis in regional lymph nodes is one of the most important prognostic factors in colorectal cancer. Nevertheless, recurrence may be expected in about 20% of lymph-node-negative patients. Detection of the lymph node(s) most likely to contain potential metastases, denoted the sentinel node(s), would be expected to contribute to the identification of patients with poor prognosis even if they have no lymph node metastases detected by routine analysis. The literature databases PubMed and EMBASE were systematically searched with combinations of the words "sentinel, lymph node, node, colon, colorectal, rectum, carcinoma and cancer". Relevant reviews and original articles among primary hits, and cross-references therein, were extracted for further study. Mapping of sentinel lymph node(s) in patients with colorectal cancer is feasible. However, a standard histopathological analysis restricted to the sentinel lymph nodes is associated with too high false negative rates to be a realistic alternative to the present routine analysis of regional lymph nodes. New prospective studies are required to clarify whether immunohistochemical or molecular biological analysis of sentinel nodes can identify patients with a poor prognosis even though they have no lymph node metastases detected by routine analysis.


Electropalatography as an adjunct to nonspeech orofacial myofunctional disorder assessments: a feasibility study.
The purpose of this study was to determine if electropalatography (EPG) would be a useful adjunct and feasible option for those conducting clinical assessments of individuals with suspected nonspeech orofacial myofunctional disorders (NSOMD). Three females (two adults, one child) were referred by their orthodontist for assessment of suspected NSOMD. Three adults and one child without NSOMD were recruited for the purpose of evaluating methodological construct, and to provide comparisons for participants with NSOMD. Using EPG, lingual-palatal timing and contact patterns of 105 saliva swallows (45 with NSOMD, 60 without NSOMD) were analyzed by compartmentalizing the sensor display and tracking the order and duration of activation. Lingual-palatal contact patterns were compared in terms of four stages: prepropulsion, propulsion, postpropulsion, release. Coding the lingual-palatal activation in an operationalized manner was a valuable adjunct for describing lingual-palatal timing and contact patterns. Participants with NSOMD showed unique lingual-palatal contact patterns that differed from the patterns of the participants without NSOMD, and from each other. EPG is a potential adjunct to the non-instrumental assessment of NSOMD. Larger scale investigations using EPG should proceed.


Towards an automatic classification of protein structural domains based on structural similarity.
Formal classification of a large collection of protein structures aids the understanding of evolutionary relationships among them. Classifications involving manual steps, such as SCOP and CATH, face the challenge of increasing volume of available structures. Automatic methods such as FSSP or Dali Domain Dictionary, yield divergent classifications, for reasons not yet fully investigated. One possible reason is that the pairwise similarity scores used in automatic classification do not adequately reflect the judgments made in manual classification. Another possibility is the difference between manual and automatic classification procedures. We explore the degree to which these two factors might affect the final classification. We use DALI, SHEBA and VAST pairwise scores on the SCOP C class domains, to investigate a variety of hierarchical clustering procedures. The constructed dendrogram is cut in a variety of ways to produce a partition, which is compared to the SCOP fold classification.Ward's method dendrograms led to partitions closest to the SCOP fold classification. Dendrogram- or tree-cutting strategies fell into four categories according to the similarity of resulting partitions to the SCOP fold partition. Two strategies which optimize similarity to SCOP, gave an average of 72% true positives rate (TPR), at a 1% false positive rate. Cutting the largest size cluster at each step gave an average of 61% TPR which was one of the best strategies not making use of prior knowledge of SCOP. Cutting the longest branch at each step produced one of the worst strategies. We also developed a method to detect irreducible differences between the best possible automatic partitions and SCOP, regardless of the cutting strategy. These differences are substantial. Visual examination of hard-to-classify proteins confirms our previous finding, that global structural similarity of domains is not the only criterion used in the SCOP classification. Different clustering procedures give rise to different levels of agreement between automatic and manual protein classifications. None of the tested procedures completely eliminates the divergence between automatic and manual protein classifications. Achieving full agreement between these two approaches would apparently require additional information.


Behavioural deviance and maternal depressive symptoms in paediatric outpatients.
A consecutive series of 95 3 to 4 year old and 43 7 to 11 year old children attending surgical, and medical outpatient clinics was studied, using questionnaires that measured behavioural deviance and had adequate reliability and validity for screening populations of children. A control group was also studied in the 7 to 11 year old sample. Depressive symptoms in the mothers of the children were determined using the Wakefield depression inventory. There was considerable behavioural deviance in children attending the outpatient clinics; 44% of the 7 to 11 year olds who were surgical outpatients had deviant scores. Thirty two per cent of mothers of medical outpatients had deviant scores on the depression inventory. These findings have relevance for the approach paediatricians take to their outpatients.


[Biochemical differences in the reactions of visual cortex neurons to deprivation].
Early visual deprivation was interferometrically shown to entail changes in the concentration and contents of the protein substance with simultaneous changes of the neurons size. The changes are statistically significant in the lamina V and not in the laminasIII and IV. The changes in each lamina are associated with definite group of the neurons the development of which is, probably, determined by visual stimuli.


[Safety of retroviral vectors in gene therapy].
Retroviral vectors are widely used in gene therapy and found to be an effective tool for the delivery of genetic constructs into cells. A unique feature of these vectors is the ability to incorporate therapeutic genes into a chromosome that ensures its passage to all progeny cells and enables to cure the diseases requiring genetic correction of dividing cells such as hematopoietic cells or skin cells. Retroviral vectors have been successfully used in gene therapy clinical trials for the treatment of 2 forms of severe combined immunodeficiencies and some other hereditary blood disorders. However, the integration of the vector into the chromosome was accompanied by genotoxicity and caused development of hematologic malignancies in several patients. Later it was shown that genotoxicity is not a general feature of retroviral vectors but it depends on many factors. In the present article we discuss safety issues concerning the use of different retroviral vectors in gene therapy. The description of modern vectors which designed to avoid the genotoxicity and other possible side effects are given.


Craniocervical arachnoid cyst in a patient with Klippel-Feil syndrome: a unique case: case report.
Klippel-Feil syndrome, or brevicollis, is a complex congenital disorder caused by the improper segmentation of the cervical vertebrae. The authors present the very rare case of a patient with Klippel-Feil syndrome who presented with an intradural arachnoid cyst at the craniocervical junction. They also examine possible factors contributing to this association. A 46-year-old woman presented with complaints of progressively worsening headaches and dizziness of 18 months' duration. She also demonstrated mild bilateral upper-extremity weakness. Magnetic resonance imaging revealed fused cervical vertebrae and a dorsal intradural arachnoid cyst at the craniocervical junction, extending down to the fourth cervical level. Because of worsening myelopathy and the presence of brainstem compression, the patient underwent surgical excision of the arachnoid cyst, which was approached via a midline posterior suboccipital/upper cervical route. An endoscope was introduced through a gap between the occiput and fused upper cervical vertebrae, and the arachnoid cyst was widely fenestrated. Postoperatively, the patient has remained symptom free for more than 2 years with evidence of good radiological decompression. The authors report a unique association between craniocervical arachnoid cyst and Klippel-Feil syndrome. To their knowledge, no other cases of this association have been reported in the literature. Arachnoid cysts should be part of the differential diagnosis in the presence of worsening myelopathic symptoms or pain in patients with Klippel-Feil syndrome.


An aberrant drawing sequence of the human figure and its relation to psychopathology.
The incidence of affective instability and antisocial behavior in patients, and familial mental illness in 14 psychiatric inpatients who drew the human figure aberrantly from the feet upwards to the head were compared with 15 psychiatric inpatients who did not draw the human figure in this manner. The incidence of the three factors of interest was significantly different in the two groups. While affective instability was not significantly different in the two groups, there were significant differences in the incidence of familial mental illness and antisocial behavior with elevations for the aberrant drawing group. Clinical vignettes are presented. This study suggests that the sequence of drawing may be a more objective datum than psychodynamic interpretations of finished products and may have some predictive usefulness in planning treatment.


Prevalence and correlates of traumatic brain injury (TBI) in older adults: results from the Well-being of the Singapore Elderly (WiSE) study.
Older adults are among the most susceptible to sustain traumatic brain injury (TBI). The study aimed to determine the (1) prevalence of TBI among older adults in Singapore, and (2) socio-demographic, lifestyle, and clinical correlates of TBI. Data were extracted from the cross-sectional, Well-being of the Singapore Elderly (WiSE) study. The study included 2,565 participants aged 60 years and above (Mean = 72.75, SD = 9.54). Information on TBI, socio-demographic, and lifestyle factors were collected using participant self-report and verified with the informant report where necessary. Disability was measured using the World Health Organization - Disability Assessment Schedule 2.0 (WHO-DAS 2.0). Data were analyzed using logistic regression analysis. The prevalence of TBI was 3.6%. Being female (vs. male) was found to be associated with decreased odds of having TBI. Having completed secondary education or lower (vs. tertiary education) was found to be associated with increased odds of having TBI. A history of fainting and diabetes were associated with the presence of TBI. Those with TBI were associated with higher disability scores on the WHO-DAS 2.0 than those without TBI. The current study provides information on the prevalence and associated factors of TBI in the older adult population in Singapore. Since TBI was associated with older adults with diabetes, they must be cautioned about fall risk. Also, given the association with disability, older adults with TBI are likely to require support and rehabilitative care to ensure good quality of life.


Conscientiousness in life course context: a commentary.
In this commentary, the common themes from the interesting articles in this special section of Developmental Psychology are considered as they illuminate the potential ontogenetic sources of the conscientiousness-well-being-longevity interconnections that have emerged in recent research. In particular, consideration is given to the changing nature of the expression of conscientiousness over the life course and the importance of the causally linked chain of developmental events associated with the sustenance of this trait from early childhood to later life. Methodological as well as conceptual issues are part of the common thematic analysis provided. In addition, several more or less neglected issues are addressed in the commentary. These include an examination of the potential roles of self-presentation social context and social aggregation in the developmentally emergent conscientiousness-well-being relationship. Further, the potential downside of the moral tinge of the conscientiousness construct is considered, as is the cultural variation in the benefits of conscientiousness to well-being. Finally, the applied significance of research on the conscientiousness-well-being link to health promotion over the life course is addressed. The commentary concludes with reflections on the transformation of 1970s trait-like conceptions as unbending tautological predictors of behavior into the situationally and developmentally nuanced conceptions of conscientiousness provided in these collected articles.


Implementing the ISO 15746 Standard for Chemical Process Optimization.
This paper proposes an approach to integrating advanced process control solutions with optimization (APC-O) solutions, within any factory, to enable more efficient production processes. Currently, vendors who provide the software applications that implement control solutions are isolated and relatively independent. Each such solution is designed to implement a specific task such as control, simulation, and optimization - and only that task. It is not uncommon for vendors to use different mathematical formalisms and modeling tools that produce different data representations and formats. Moreover, instead of being modeled uniformly only once, the same knowledge is often modeled multiple times - each time using a different, specialized abstraction. As a result, it is extremely difficult to integrate optimization with advanced process control. We believe that a recent standard, International Organization for Standardization (ISO) 15746, describes a data model that can facilitate that integration. In this paper, we demonstrate a novel method of integrating advanced process control using ISO 15746 with numerical optimization. The demonstration is based on a chemical-process-optimization problem, which resides at level 2 of the International Society of Automation (ISA) 95 architecture. The inputs to that optimization problem, which are captured in the ISO 15746 data model, come in two forms: goals from level 3 and feedback from level 1. We map these inputs, using this data model, to a population of a meta-model of the optimization problem for a chemical process. Serialization of the metamodel population provides input to a numerical optimization code of the optimization problem. The results of this integrated process, which is automated, provide the solution to the originally selected, level 2 optimization problem.


Challenges and Opportunities Modeling the Dynamic Tumor Matrisome.
We need novel strategies to target the complexity of cancer and, particularly, of metastatic disease. As an example of this complexity, certain tissues are particularly hospitable environments for metastases, whereas others do not contain fertile microenvironments to support cancer cell growth. Continuing evidence that the extracellular matrix (ECM) of tissues is one of a host of factors necessary to support cancer cell growth at both primary and secondary tissue sites is emerging. Research on cancer metastasis has largely been focused on the molecular adaptations of tumor cells in various cytokine and growth factor environments on 2-dimensional tissue culture polystyrene plates. Intravital imaging, conversely, has transformed our ability to watch, in real time, tumor cell invasion, intravasation, extravasation, and growth. Because the interstitial ECM that supports all cells in the tumor microenvironment changes over time scales outside the possible window of typical intravital imaging, bioengineers are continuously developing both simple and sophisticated in vitro controlled environments to study tumor (and other) cell interactions with this matrix. In this perspective, we focus on the cellular unit responsible for upholding the pathologic homeostasis of tumor-bearing organs, cancer-associated fibroblasts (CAFs), and their self-generated ECM. The latter, together with tumoral and other cell secreted factors, constitute the "tumor matrisome". We share the challenges and opportunities for modeling this dynamic CAF/ECM unit, the tools and techniques available, and how the tumor matrisome is remodeled (e.g., via ECM proteases). We posit that increasing information on tumor matrisome dynamics may lead the field to alternative strategies for personalized medicine outside genomics.


The isolation of neurophysin-I and-II from bovine pituitary neurosecretory granules separated on a large scale from other subcellular organelles. Demonstration of slow equilibration of neurosecretory granules during centrifugation in a sucrose density gradient.
1. An improved procedure for the isolation of neurosecretory granules from the posterior lobe of the bovine pituitary gland is described. 2. Of the total oxytocic and pressor activities present in the original tissue 80% was sedimentable. 3. The granules were separated from mitochondria by prolonged centrifugation in a sucrose density gradient. During a sedimentation period of 5hr. the granules moved progressively into denser regions of the gradient and the mitochondria remained at the top. 4. The biological activities of the granules were measured: the oxytocic activity was 11.56+/-1.63 and the pressor activity was 15.60+/-3.91 units/mg. of protein. 5. A protein was isolated from a lysate of granules prepared from 40 pituitary glands. Amino acid analysis showed that it consisted of a mixture of neurophysin-I and neurophysin-II in equal proportions. It accounted for 60% of the soluble granule protein and for 50% of the total granule protein. 6. The neurophysins present in the granules are associated with 19.1 units of oxytocic and 21.1 units of pressor activity/mg. of protein. 7. Starch-gel electrophoresis revealed the presence of both neurophysins in extracts of 15 pituitary glands studied individually. 8. We conclude that the polypeptide hormones, oxytocin and [8-arginine]-vasopressin, are normally closely associated with the two neurophysins within neurosecretory granules of the pituitary gland.


Threshold differences for interaural time delays carried by double vowels.
Experimental measurements were made of threshold interaural time differences (ITDs) for a "target" vowel presented simultaneously with a fixed-ITD "distracter" vowel. Three double-vowel pairs were used, comprising an "er" (/e/) together with either an "ai," "ar," or "oo" (respectively, /e/, /c/, and /u/). Threshold ITDs were found to be larger for the target vowel when it was part of a double-vowel pair than in control conditions in which it was presented alone. The effect size depended upon the choice of target vowel and distracter vowel, the level of the target relative to the distracter, and whether the two vowels had the same or different fundamental frequencies. The experiment was analyzed using a multichannel modification of Heller and Trahiotis' [J. Acoust. Soc. Am. 99, 3632-3637 (1996)] model, which used a weighted combination of the detectabilities of the ITD of the target and the distracter. It gave predictions consistent with the observed effects of level and with some of the effects of the choice of target vowel, but it could not describe the effect of the target-distracter differences in fundamental frequency. It was found that a single-channel version of the model, in which the chosen channel was allowed to depend upon fundamental frequency (which could be derived using a monaural autocorrelation model) did give a set of predictions in qualitative accord with the data.


Process evaluation: how it works.
Process evaluation helps us to understand the planning process. This predominantly qualitative approach explains how and why decisions are made and activities undertaken. The focus includes feelings and perceptions of program staff. The evaluator's ability to interpret and longitudinally summarize the experience of program staff and community members is critical. Techniques discussed include participant observation, content analysis, situational analysis, in-house surveys, and interviews. By combining sources and methods, a fuller picture of the process is revealed.


[Isolation and characteristics of highly-purified phenylalanine hydroxylase from the rat liver].
Electrophoretically homogenous phenylalanine hydroxylase was isolated from rat liver tissue using a number of methods involving affinity chromatography on phenyl-Sepharose, ion exchange chromatography on DEAE-Toyoperle and hydrophobic rechromatography on phenyl-Sepharose. Molecular mass of the enzyme was equal to 50,000 Da and Km = 4.4 X 10(-3) M with phenylalanine as a substrate. The enzymatic preparations were used for immunization of rabbits and the monospecific antiserum towards phenylalanine hydroxylase was produced.


The thymus plays a role in oral tolerance in experimental autoimmune encephalomyelitis.
The oral administration of myelin proteins has been used for the successful prevention and treatment of experimental autoimmune encephalomyelitis (EAE). We questioned whether the thymus was involved in oral tolerance. In this study, euthymic myelin basic protein (MBP) TCR transgenic mice are protected from EAE when fed MBP but are not protected when thymectomized. Similarly, in a cell transfer system, T cell responses to OVA measured in vivo were suppressed significantly only in the OVA-fed euthymic mice but not in the thymectomized mice. We observed that the absence of the thymus dramatically enhanced the Th1 response. We explored three alternatives to determine the role of the thymus in oral tolerance: 1) as a site for the induction of regulatory T cells; 2) a site for deletion of autoreactive T cells; or 3) a site for the dissemination of naive T cells. We found that Foxp3(+)CD4(+)CD25(+) T cells are increased in the periphery but not in the thymus after Ag feeding. These CD4(+)CD25(+) T cells also express glucocorticoid-induced TNFR and intracellular CTLA4 and suppress Ag-specific proliferation of CD4(+)CD25(-) cells in vitro. The thymus also plays a role in deletion of autoreactive T cells in the periphery following orally administered MBP. However, thymectomy does not result in homeostatic proliferation and the generation of memory cells in this system. Overall, the oral administration of MBP has a profound effect on systemic immune responses, mediated largely by the generation of regulatory T cells that act to prevent or suppress EAE.


Ethanolic extract of Taheebo attenuates increase in body weight and fatty liver in mice fed a high-fat diet.
We evaluated whether intake of an ethanolic extract of Taheebo (TBE) from Tabebuia avellanedae protects against body weight increase and fat accumulation in mice with high-fat diet (HFD)-induced obesity. Four-week old male C57BL/6 mice were fed a HFD (25% fat, w/w) for 11 weeks. The diet of control (HFD) mice was supplemented with vehicle (0.5% sodium carboxymethyl cellulose by gavage); the diet of experimental (TBE) mice was supplemented with TBE (150 mg/kg body weight/day by gavage). Mice administered TBE had significantly reduced body weight gain, fat accumulation in the liver, and fat pad weight, compared to HFD mice. Reduced hypertrophy of fat cells was also observed in TBE mice. Mice administered TBE also showed significantly lower serum levels of triglycerides, insulin, and leptin. Lipid profiles and levels of mRNAs and proteins related to lipid metabolism were determined in liver and white adipose tissue of the mice. Expression of mRNA and proteins related to lipogenesis were decreased in TBE-administered mice compared to mice fed HFD alone. These results suggest that TBE inhibits obesity and fat accumulation by regulation of gene expression related to lipid metabolism in HFD-induced obesity in mice.


Heparan Sulfate and Chondroitin Sulfate Glycosaminoglycans Are Targeted by Bleomycin in Cancer Cells.
Bleomycin is a clinically used anti-cancer drug that produces DNA breaks once inside of cells. However, bleomycin is a positively charged molecule and cannot get inside of cells by free diffusion. We previously reported that the cell surface negatively charged glycosaminoglycans (GAGs) may be involved in the cellular uptake of bleomycin. We also observed that a class of positively charged small molecules has Golgi localization once inside of the cells. We therefore hypothesized that bleomycin might perturb Golgi-operated GAG biosynthesis. We used stable isotope labeling coupled with LC/MS analysis of GAG disaccharides simultaneously from bleomycin-treated and non-treated cancer cells. To further understand the cytotoxicity of bleomycin and its relationship to GAGs, we used sodium chlorate to inhibit GAG sulfation and commercially available GAGs to compete for cell surface GAG/bleomycin interactions in seven cell lines including CHO745 defective in both heparan sulfate and chondroitin sulfate biosynthesis. we discovered that heparan sulfate GAG was significantly undersulfated and the quantity and disaccharide compositions of GAGs were changed in bleomycin-treated cells in a concentration- and time-dependent manner. We revealed that bleomycin-induced cytotoxicity was directly related to cell surface GAGs. GAGs were targeted by bleomycin both at cell surface and at Golgi. Thus, GAGs might be the biological relevant molecules that might be related to the bleomycin-induced fibrosis in certain cancer patients, a severe side effect with largely unknown molecular mechanism.


Quantitative and functional analysis of PDC-E2-specific autoreactive cytotoxic T lymphocytes in primary biliary cirrhosis.
While the pathologic mechanisms responsible for organ-specific tissue damage in primary biliary cirrhosis (PBC) remain an enigma, it has been suggested that the pathology is mediated by autoreactive T cells infiltrating the intrahepatic bile ducts. Previously, we have documented that there is 100-fold enrichment in the frequency of CD4(+) autoreactive T cells in the liver that are specific for peptides encoded by the E2 components of the pyruvate dehydrogenase complexes (PDC-E2). We have also recently characterized the first MHC class I-restricted epitope for PDC-E2, namely amino acid 159-167, a region very similar to the epitope recognized by MHC class II-restricted CD4(+) cells and by autoantibodies. The effector functions of these PDC-E2(159-167)-specific CD8(+) cytotoxic T lymphocytes (CTLs) are not well understood. We have taken advantage of tetramer technology and report herein that there is tenfold increase in the frequency of PDC-E2(159-167)-specific CTLs in the liver as compared with the blood in PBC. In addition, the precursor frequency of the CTLs in blood was significantly higher in early-stage PBC. Of interest was the fact that, upon stimulation with the peptide, the response of PDC-E2(159-167) tetramer-positive cells is heterogeneous with respect to IFN-gamma synthesis. These data, we believe for the first time, document the enrichment of autoantigen-specific CD8(+) T cells in the PBC liver, suggesting that CD8(+) T cells play a significant role in the immunopathogenesis of PBC.


Dyadic relationship conflict, gender, and mortality in urban hemodialysis patients.
The effects of dyadic satisfaction and conflict have not been well defined in the hemodialysis (HD) population. The aim of this study was to determine whether the perception of decreased dyadic satisfaction was associated with mortality in patients treated with HD, and if so, whether there were different relationships between risk factors, and differential outcomes in men and women. A total of 174 HD patients, primarily African-Americans, involved in dyadic relationships for more than 6 mo had indices of dyadic satisfaction, depression, perception of illness effects, social support, behavioral compliance with the dialysis prescription, and plasma interleukin-1 (IL-1) and beta-endorphin levels measured. Cox proportional hazards models assessed relative mortality risks. Patients' dyadic satisfaction scores correlated with beta-endorphin levels. There was no correlation of IL-1 or beta-endorphin with any psychosocial or behavioral compliance measure in the group as a whole. Correlations between psychosocial, medical, and neuroimmunologic variables were different in men and women. For women, dyadic satisfaction correlated with beta-endorphin levels, depression, and perception of illness. Women with higher dyadic satisfaction and decreased dyadic conflict were at decreased mortality risk, but dyadic adjustment indices were unassociated with differential survival in the larger group of men. Correlations between neuroendocrine and immune markers are different in African-American male and female HD patients. Greater dyadic satisfaction and lower dyadic conflict are independently associated with decreased mortality in female African-American HD patients, of the same order of magnitude as medical risk factors. Such effects may be attributable to a relationship between dyadic satisfaction and conflict and health-related behaviors, or through an effect on neuroendocrine or immunologic status.


Comparison of economic impact of chronic prostatitis/chronic pelvic pain syndrome and interstitial cystitis/painful bladder syndrome.
To perform a comparison of the economic impact of chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) and interstitial cystitis/painful bladder syndrome (IC/PBS) because limited information is available. Furthermore, no direct comparisons of the costs of these 2 conditions have been performed. Such a comparison is relevant because the distinction between the 2 conditions is not always clear. We recruited 62 men with CP/CPPS and 43 women with IC/PBS from a tertiary care outpatient urology clinic. Information about hospitalizations, laboratory tests, physician visits, telephone calls, medication use, and lost productivity was obtained from written questionnaires. Direct medical cost estimates were determined from hospital cost accounting data, the 2005 Physician Fee Schedule Book, and the 2005 Redbook for pharmaceuticals. Indirect costs were determined from patient-reported annual income and patient-reported hours lost from work during the most recent 3-month period. Using Medicare rates, the annualized direct costs per person were $3631 for IC/PBS and $3017 for CP/CPPS. Using non-Medicare rates for outpatient visits and tests/procedures, the annual per person costs increased substantially to $7043 for IC/PBS and $6534 for CP/CPPS. Sixteen patients with CP/CPPS (26%) and 8 with IC/PBS (19%) reported lost wages as a result of their condition in the previous 3 months. Both CP/CPPS and IC/PBS have very similar and substantial direct and indirect costs. The greater costs reflected by the non-Medicare rates may more accurately reflect the true costs, given that a large proportion of these patients were <65 years old.


THz streak camera method for synchronous arrival time measurement of two-color hard X-ray FEL pulses.
The two-color operation of free electron laser (FEL) facilities allows the delivery of two FEL pulses with different energies, which opens new possibilities for user experiments. Measuring the arrival time of both FEL pulses relative to the external experimental laser and to each other improves the temporal resolution of the experiments using the two-color FEL beam and helps to monitor the performance of the machine itself. This work reports on the first simultaneous measurement of the arrival times of two hard X-ray FEL pulses with the THz streak camera. Measuring the arrival time of the two FEL pulses, the relative delay between them was calculated and compared to the set values. Furthermore, we present the first comparison of the THz streak camera method to the method of FEL induced transient transmission. The results indicate a good agreement between the two methods.


Signal-transduction-pathway-specific desensitization of expression of orphan nuclear receptor TIS1.
Induction of many primary-response genes by a variety of stimuli occurs in a transient manner. The precise mechanism responsible for these transient kinetics is not completely understood. We report here that the orphan nuclear receptor, TIS1, which is a potent sequence-specific transcription factor, was transiently induced by the adrenergic agonist isoprenaline in the C2C12 skeletal-muscle cell line. Moreover, we showed that the rapid decline in mRNA level after peak induction was due in part to a specific desensitization of the isoprenaline-mediated induction pathway. Desensitization of the induction response presumably occurred at the level of the receptor, as agents that either bypass the adrenergic receptor or activate alternative signalling pathways were able to induce TIS1 expression in the desensitized cells. However, stimulation by agents that directly activate intracellular enzymes also resulted in the signal-transduction-pathway-specific desensitization of TIS1 inducibility. Our results suggest that the pathway-specific nature of the desensitization process may be important for directing an integrated response to multiple physiological stimuli.


The Clostridium difficile exosporium cysteine (CdeC)-rich protein is required for exosporium morphogenesis and coat assembly.
Clostridium difficile is an important nosocomial pathogen that has become a major cause of antibiotic-associated diarrhea. There is a general consensus that C. difficile spores play an important role in C. difficile pathogenesis, contributing to infection, persistence, and transmission. Evidence has demonstrated that C. difficile spores have an outermost layer, termed the exosporium, that plays some role in adherence to intestinal epithelial cells. Recently, the protein encoded by CD1067 was shown to be present in trypsin-exosporium extracts of C. difficile 630 spores. In this study, we renamed the CD1067 protein Clostridium difficile exosporium cysteine-rich protein (CdeC) and characterized its role in the structure and properties of C. difficile spores. CdeC is expressed under sporulation conditions and localizes to the C. difficile spore. Through the construction of an ΔcdeC isogenic knockout mutant derivative of C. difficile strain R20291, we demonstrated that (i) the distinctive nap layer is largely missing in ΔcdeC spores; (ii) CdeC is localized in the exosporium-like layer and is accessible to IgGs; (iii) ΔcdeC spores were more sensitive to lysozyme, ethanol, and heat treatment than wild-type spores; and (iv) despite the almost complete absence of the exosporium layer, ΔcdeC spores adhered at higher levels than wild-type spores to intestinal epithelium cell lines (i.e., HT-29 and Caco-2 cells). Collectively, these results indicate that CdeC is essential for exosporium morphogenesis and the correct assembly of the spore coat of C. difficile.


Brain metastases in patients with EGFR-mutated or ALK-rearranged non-small-cell lung cancers.
Brain metastases (BM) are common in non-small-cell lung cancer (NSCLC). However, the baseline incidence and evolution of BM over time in oncogene-driven NSCLCs are seldom reported. In this study, we evaluated the frequency of BM in patients with epidermal growth factor receptor (EGFR)-mutated or anaplastic lymphoma kinase (ALK)-rearranged NSCLC. The presence of BM, clinicopathologic data, and tumor genotype were retrospectively compiled and analyzed from a cohort of 381 patients. We identified 86 EGFR-mutated (90.7% with metastatic disease; 85.9% received an EGFR inhibitor) and 23 ALK-rearranged (91.3% with metastatic disease; 85.7% received an ALK inhibitor) NSCLCs. BM were present in 24.4% of EGFR-mutated and 23.8% of ALK-rearranged NSCLCs at the time of diagnosis of advanced disease. This study did not demonstrate a difference in the cumulative incidence of BM over time between the two cohorts (EGFR/ALK cohort competing risk regression [CRR] coefficient of 0.78 [95% CI 0.44-1.39], p=0.41). In still living patients with advanced EGFR-mutated NSCLC, 34.2% had BM at 1 year, 38.4% at 2 years, 46.7% at 3 years, 48.7% at 4 years, and 52.9% at 5 years. In still living patients with advanced ALK-rearranged NSCLC, 23.8% had BM at 1 year, 45.5% at 2 years, and 58.4% at 3 years. BM are frequent in advanced EGFR-mutated or ALK-rearranged NSCLCs, with an estimated >45% of patients with CNS involvement by three years of survival with the use of targeted therapies. These data point toward the CNS as an important unmet clinical need in the evolving schema for personalized care in NSCLC.


Identifying an Appropriate Area to Facilitate the Cardiorespiratory Measurement during Sleep Monitoring.
Measuring cardiorespiratory parameters in sleep, using non-contact sensors and the Ballistocardiography technique has received much attention due to the low-cost, unobtrusive, and non-invasive method. Designing a user-friendly, simple-to-use, and easy-to-deployment preserving less error-prone remains open and challenging due to the complex morphology of the signal. In this work, using four forcesensitive resistor sensors, we conducted a study by designing four distributions of sensors, in order to simplify the complexity of the system by identifying the region of interest for heartbeat and respiration measurement. The sensors are deployed under the mattress and attached to the bed frame without any interference with the subjects. The four distributions are combined in two linear horizontal, one linear vertical, and one square, covering the influencing region in cardiorespiratory activities. We recruited 4 subjects and acquired data in four regular sleeping positions, each for a duration of 80 seconds. The signal processing was performed using discrete wavelet transform bior 3.9 and smooth level of 4 as well as bandpass filtering. The results indicate that we have achieved the mean absolute error of 2.35 and 4.34 for respiration and heartbeat, respectively. The results recommend the efficiency of a triangleshaped structure of three sensors for measuring heartbeat and respiration parameters in all four regular sleeping positions.


Neurotensin is a proinflammatory neuropeptide in colonic inflammation.
The neuropeptide neurotensin mediates several intestinal functions, including chloride secretion, motility, and cellular growth. However, whether this peptide participates in intestinal inflammation is not known. Toxin A, an enterotoxin from Clostridium difficile, mediates pseudomembranous colitis in humans. In animal models, toxin A causes an acute inflammatory response characterized by activation of sensory neurons and intestinal nerves and immune cells of the lamina propria. Here we show that neurotensin and its receptor are elevated in the rat colonic mucosa following toxin A administration. Pretreatment of rats with the neurotensin receptor antagonist SR-48, 692 inhibits toxin A-induced changes in colonic secretion, mucosal permeability, and histologic damage. Exposure of colonic explants to toxin A or neurotensin causes mast cell degranulation, which is inhibited by SR-48,692. Because substance P was previously shown to mediate mast cell activation, we examined whether substance P is involved in neurotensin-induced mast cell degranulation. Our results show that neurotensin-induced mast cell degranulation in colonic explants is inhibited by the substance P (neurokinin-1) receptor antagonist CP-96,345, indicating that colonic mast activation in response to neurotensin involves release of substance P. We conclude that neurotensin plays a key role in the pathogenesis of C. difficile-induced colonic inflammation and mast cell activation.


Specific recognition of Leishmania major poly-beta-galactosyl epitopes by galectin-9: possible implication of galectin-9 in interaction between L. major and host cells.
Leishmania parasites are the causative agents of leishmaniasis, manifesting itself in a species-specific manner. The glycan epitopes on the parasite are suggested to be involved in the Leishmania pathogenesis. One of such established species-unique glycan structures is the poly-beta-galactosyl epitope (Galbeta1-3)n found on L. major, which can develop cutaneous infections with strong inflammatory responses. Interestingly, the polygalactosyl epitope is also suggested to be involved in the development of the parasites in its host vector, sand fly. Thus, the recognition of the galactosyl epitope by lectins expressed in host or sand fly should be implicated in the species-specific manifestations of leishmaniasis and in the parasite life cycle, respectively. We recently reported that one host beta-galactoside-binding protein, galectin-3, can distinguish L. major from the other species through its binding to the poly-beta-galactosyl epitope, proposing a role for galectin-3 as an immunomodulator that could influence the L. major-specific immune responses in leishmaniasis. Here we report that galectin-9 can also recognize L. major by binding to the L. major-specific polygalactosyl epitope. Frontal affinity analysis with different lengths of poly-beta-galactosyllactose revealed that the galectin-9 affinity for polygalactose was enhanced in proportion to the number of Galbeta1-3 units present. Even though both galectins have comparable affinities toward the polygalactosyl epitopes, only galectin-9 can promote the interaction between L. major and macrophages, suggesting distinctive roles for the galectins in the L. major-specific development of leishmaniasis in the host.


Factors involved in specific transcription by mammalian RNA polymerase II: role of transcription factors IIA, IID, and IIB during formation of a transcription-competent complex.
Human transcription factor TFIID, the TATA-binding protein, was partially purified to a form capable of associating stably with the TATA motif of the adenovirus major late promoter. Binding of the human and yeast TFIID to the TATA motif was stimulated by TFIIA. TFIIA is an integral part of a complex capable of binding other transcription factors. A complex formed with human TFIID and TFIIA (DA complex) was specifically recognized by TFIIB. We found that TFIIB activity was contained in a single polypeptide of 32 kDa and that this polypeptide participated in transcription and was capable of binding to the DA complex to form the DAB complex. Formation of the DAB complex required TFIIA, TFIID, and sequences downstream of the transcriptional start site; however, the DA complex could be formed on an oligonucleotide containing only the adenovirus major late promoter TATA motif. Using anti-TFIIB antibodies and reagents that affect the stability of a transcription-competent complex, we found that yeast and human TFIID yielded DAB complexes with different stabilities.


Outcomes of Baerveldt Glaucoma Drainage Devices in Pediatric Eyes.
In children, Baerveldt implants showed 84% success at 1 year, but decreased to 32% at 8 years. Age, race, and glaucoma type were not risk factors for failure. Concurrent intraocular surgery was associated with complications. Evaluate success and risk factors for failure and complications of Baerveldt glaucoma implants in children. Retrospective case series of children who underwent Baerveldt implant placement (2012-2019 by single surgeon) with ≥1 year follow-up. Ocular examination and surgical details were collected. Failure defined as intraocular pressure (IOP) <5 mm Hg or >21 mm Hg for 2 consecutive visits, need for IOP related surgery, or visually significant complication. One hundred-six eyes of 76 patients underwent 110 Baerveldt placement at median 6.4 years. Baerveldt placement was combined with additional procedures in 49% with vitrectomy most common (30%). Success of first Baerveldt (per patient) was 64% at final follow-up (median 4.7 y). One-, 5-, and 8-year survival rates were 84%, 60%, and 32%, respectively. There was no difference (P=0.97) in survival between first Baerveldt and all Baerveldt surgeries. Failure of first Baerveldt was not associated with sex, age, ethnicity, prior IOP-lowering surgery, concurrent intraocular surgery, or glaucoma type. Complications occurred in 14% and were associated with concurrent surgery. Twenty-six percent required additional IOP-lowering surgery. At final follow-up, IOP and glaucoma medications were significantly decreased (P<0.0001). Eyes underwent an average of 3.8±2.3 ocular surgeries and 3.0±2.0 glaucoma surgeries. Baerveldt implants showed good success initially, but survival rates declined over time. No risk factors for failure of first implanted Baerveldt were identified. Concurrent surgery was associated with complications. Majority of eyes required multiple surgeries to achieve IOP control and preserve vision.


The influence of abdominal visceral fat on inflammatory pathways and mortality risk in obstructive lung disease.
Low-grade systemic inflammation, particularly elevated IL-6, predicts mortality in chronic obstructive pulmonary disease (COPD). Although altered body composition, especially increased visceral fat (VF) mass, could be a significant contributor to low-grade systemic inflammation, this remains unexplored in COPD. The objective was to investigate COPD-specific effects on VF and plasma adipocytokines and their predictive value for mortality. Within the Health, Aging, and Body Composition (Health ABC) Study, an observational study in community-dwelling older persons, we used propensity scores to match n = 729 persons with normal lung function to n = 243 persons with obstructive lung disease (OLD; defined as the ratio of forced expiratory volume in 1 s to forced vital capacity < lower limit of normal). Matching was based on age, sex, race, clinic site, BMI, and smoking status. Within this well-balanced match, we compared computed tomography-acquired visceral fat area (VFA) and plasma adipocytokines, analyzed independent associations of VFA and OLD status on plasma adipocytokines, and studied their predictive value for 9.4-y mortality. Whereas whole-body fat mass was comparable between groups, persons with OLD had increased VFA and higher plasma IL-6, adiponectin, and plasminogen activator inhibitor 1 (PAI-1). Both OLD status and VFA were independently positively associated with IL-6. Adiponectin was positively associated with OLD status but negatively associated with VFA. PAI-1 was no longer associated with OLD status after VFA was accounted for. Participants with OLD had increased risk of all-cause, respiratory, and cardiovascular mortality, of which IL-6 was identified as an independent predictor. Our data suggest that excessive abdominal visceral fat contributes to increased plasma IL-6, which, in turn, is strongly associated with all-cause and cause-specific mortality in older persons with OLD.


How Do Families Experience and Interact with CPS?
The lived experiences of child protective services (CPS)-involved parents is rarely considered from a social justice perspective. Parents and children endure the oversight of the child welfare system in myriad ways, and these experiences usually vary based on race, ethnicity, and socioeconomic status. This article explores how CPS interactions affect family dynamics and well-being and how family members view their experiences with CPS, including their sense of autonomy and empowerment. I focus on the inherent power dynamics between CPS workers and parents, race and ethnicity, and family. I highlight the perspectives of parents and their intended (rather than unintentional) parental behaviors (e.g., providing healthy food choices) to understand ways in which their socioecological contexts impact the well-being of their children. I report results of a pilot study designed to enhance the voices of parents in the literature and provide recommendations for policy and practice that inform innovative solutions to better support CPS-involved families.


Antioxidants protect proteins' anchorage to the bilayer by improving plasma membrane integrity of ram spermatozoa during liquid preservation in a soya lecithin-based diluent.
Antioxidants are known to prevent the reactive oxygen species (ROS)-mediated peroxidative damage to the membrane lipids during hypothermic storage of mammalian spermatozoa. We hypothesized here that ROS also affect the lipid-protein interactions, thereby diminishing the membrane's integrity and proteins' anchorage to the bilayer. Antioxidants prevent these damages by scavenging the ROS. Ejaculates from Patanwadi rams were pooled after subjective evaluation and centrifuged using Percoll® . Sperm pellet was resuspended in soya lecithin-Tris-fructose diluent (400 × 106 cells/ml) containing either antioxidants (100 IU/ml catalase + 10 mM reduced glutathione) or no antioxidant. Aliquots were chilled to 5°C in a cabinet and stored in a refrigerator at 3-5°C for 72 hr. Sperm motility, viability, lipid peroxidation (LPO) and hypo-osmotic swelling test (HOST) were performed at 0, 24, 48 and 72 hr. Sperm proteins extracted with 0.5% Triton X-100 were resolved by SDS-PAGE and quantified using Quantity One software (Bio-Rad, USA). The rapid motility, linearity and straight-line velocity (VSL) were found significantly (p < .05) higher in the antioxidant-treated group compared to the control at 48 hr of storage. Sperm viability was found comparable between the groups. Higher HOST response and lower LPO were found in the antioxidant-treated sample compared to the control both at 48 and at 72 hr. Overall, the proteins P1 (106.09 kDa), P2 (87.00 kDa) and P4 (51.14 kDa) were lower (p < .05) in the sperm extract of antioxidant-treated group compared to the control. The content of P4 (51.14 kDa) in sperm extract was found to increase (p < .05) earlier (48 vs. 72 hr) in the control group compared to the antioxidant-treated group. Altogether, the results suggested that antioxidants reduced LPO in spermatozoa, resulting in higher sperm motility, plasma membrane integrity and protection of proteins' anchorage to the plasma membrane at 48 and 72 hr of storage.


Long-term outcome in open spina bifida.
Doctors need reliable data on outcome in order to help parents faced with difficult decisions about termination of an affected pregnancy or treatment after birth. To determine survival, health and lifestyle at the mean age of 30 years in a complete cohort of adults born with open spina bifida. Prospective cohort study. Well-documented cohort of 117 consecutive cases of open spina bifida whose backs were closed non-selectively at birth between 1963 and 1971. Survivors (age range = 26 to 33 years) were surveyed by postal questionnaire and telephone interview. The main outcome measures were the health, independence and lifestyle of the survivors in terms of living in the community, driving a car and working in open employment. Ascertainment was 100%. Sixty (51%) had died, mainly the most disabled. Of the 57 survivors, 84% had a cerebrospinal fluid (CSF) shunt, 70% had an IQ of 80 or more, 37% lived independently in the community, 39% drove a car, 30% could walk more than 50 metres and 26% were in open employment. However one-third (19) still needed daily care, three were on respiratory support, two were blind, two had diabetes mellitus, and one was on dialysis. Mortality, disability and achievement reflected the neurological deficit that had been recorded in infancy in terms of sensory level. Attainment and independence were reduced in those who had needed revision of CSF shunt. The survivors in this unselected cohort showed a wide range of outcome from apparent normality to very severe disability. This reflected both the extent of their original neurological deficit and events in the history of their CSF shunt.


Gene therapy approaches for GM1 gangliosidosis: Focus on animal and cellular studies.
One of the most important inherited metabolic disorders is GM1 gangliosidosis, which is a progressive neurological disorder. The main cause of this disease is a genetic defect in the enzyme β-galactosidase due to a mutation in the glb1 gene. Lack of this enzyme in cells (especially neurons) leads to the accumulation of ganglioside substrate in nerve tissues, followed by three clinical forms of GM1 disease (neonatal, juvenile, and adult variants). Genetically, many mutations occur in the exons of the glb1 gene, such as exons 2, 6, 15, and 16, so the most common ones reported in scientific studies include missense/nonsense mutations. Therefore, many studies have examined the genotype-phenotype relationships of this disease and subsequently using gene therapy techniques have been able to reduce the complications of the disease and alleviate the signs and symptoms of the disease. In this regard, the present article reviews the general features of GM1 gangliosidosis and its mutations, as well as gene therapy studies and animal and human models of the disease.


"Salt and Pepper Endothelium" Recurring After Descemet Membrane Endothelial Keratoplasty.
To describe the presence of "salt and pepper endothelium", that is, typical cellular inclusion bodies in a patient with Fuchs endothelial corneal dystrophy (FECD), that recurred in the donor corneal endothelial cells after Descemet membrane endothelial keratoplasty (DMEK). A 76-year-old man underwent DMEK for FECD in his left eye. Routine specular microscopy imaging, best-corrected visual acuity measurements, and pachymetry measurements were performed before and after surgery. Besides large guttae indicating FECD, preoperative specular microscopy images showed variable-sized dark cellular inclusion bodies in the endothelial cells. One month after DMEK, donor endothelial cells appeared normal; however, at 3 months, the typical inclusion bodies reappeared and progressed slowly within a 4-year follow-up period. Both best-corrected visual acuity and pachymetry were stable throughout the study period. "Salt and pepper endothelium" recurred after the host tissue was exchanged by donor Descemet membrane, that is, a DMEK graft. These changes may indicate that either the donor corneal endothelial cell morphology is modified by adjacent tissue structures or that it is completely replaced by recipient endothelium within the first months after surgery.


Multidrug-resistant tuberculosis in admitted patients at a tertiary referral hospital of Bangladesh.
This study was set out to investigate the magnitude, patterns and molecular characterization of drug-resistant Mycobacterium tuberculosis strains at a tertiary referral hospital in Bangladesh. Pulmonary tuberculosis (TB) patients admitted at National Institute of Diseases of the Chest and Hospital from February 2002 to September 2005 with or without previous history of TB and/or other complications were randomly interviewed. Among 265 participants enrolled, M. tuberculosis isolates from 189 patients were finally tested for susceptibility to rifampicin (RMP), isoniazid (INH), ethambutol (ETM) and streptomycin (STM). Genotyping of M. tuberculosis was done using deletion analysis and spoligotyping. Eighty-eight percent (n = 167) of the patients had history of previous anti-TB treatment while the remaining 12% were new TB cases. Of the 189 isolates, 9% were fully susceptible to the first line anti-TB drugs and 73.5% were multi-drug resistant TB. Other susceptibility results showed 79.4%, 77.2%, 76.7% and 78.8% resistance to INH, RMP, ETM and STM respectively. Multi-drug resistance was significantly higher among the 130 (78%) patients with previous history of anti-tuberculosis treatment (95% confidence interval, p = 0.001). Among the 189 analyzed isolates, 69% were classified as "modern" M. tuberculosis strains (i.e. TbD1- strains, lacking the M. tuberculosis-deletion region TbD1), whereas the remaining 31% were found to belong to the "ancestal" TbD1+ M. tuberculosis lineages. One hundred and five different spoligotype patterns were identified in which 16 clusters contained 100 strains and 89 strains had unique pattern. Strains with a spoligotype characteristic for the "Beijing" cluster were predominant (19%) and most of these strains (75%) were multi-drug resistant (MDR). A high level of drug resistance observed among the re-treatment patients poses a threat of transmission of resistant strains to susceptible persons in the community. Proper counseling of patients and attention towards the completion of the anti-TB treatment is needed.


Clinical characterization and genetic analysis of Korean patients with X-linked Charcot-Marie-Tooth disease type 1.
Mutations in the gap junction protein beta 1 gene (GJB1) cause X-linked Charcot-Marie-Tooth disease type 1 (CMTX1). CMTX1 is representative of the intermediate type of CMT, having both demyelinating and axonal neuropathic features. We analyzed the clinical and genetic characterization of 128 patients with CMTX1 from 63 unrelated families. Genetic analysis revealed a total of 43 mutations including 6 novel mutations. Ten mutations were found from two or more unrelated families. p.V95M was most frequently observed. The frequency of CMTX1 was 9.6% of total Korean CMT family and was 14.8% when calculated within genetically identified cases. Among 67 male and 61 female patients, 22 females were asymptomatic. A high-arched foot, ataxia, and tremor were observed in 87%, 41%, and 35% of the patients, respectively. In the male patients, functional disability scale, CMT neuropathy score, and compound muscle action potential of the median/ulnar nerves were more severely affected than in the female patients. This study provides a comprehensive summary of the clinical features and spectrum of GJB1 gene mutations in Korean CMTX1 patients.


Conditional deletion of Ccm2 causes hemorrhage in the adult brain: a mouse model of human cerebral cavernous malformations.
Cerebral cavernous malformations (CCM) are irregularly shaped and enlarged capillaries in the brain that are prone to hemorrhage, resulting in headaches, seizures, strokes and even death in patients. The disease affects up to 0.5% of the population and the inherited form has been linked to mutations in one of three genetic loci, CCM1, CCM2 and CCM3. To understand the pathophysiology underlying the vascular lesions in CCM, it is critical to develop a reproducible mouse genetic model of this disease. Here, we report that limited conditional ablation of Ccm2 in young adult mice induces observable neurological dysfunction and reproducibly results in brain hemorrhages whose appearance is highly reminiscent of the lesions observed in human CCM patients. We first demonstrate that conventional or endothelial-specific deletion of Ccm2 leads to fatal cardiovascular defects during embryogenesis, including insufficient vascular lumen formation as well as defective arteriogenesis and heart malformation. These findings confirm and extend prior studies. We then demonstrate that the inducible deletion of Ccm2 in adult mice recapitulates the CCM-like brain lesions in humans; the lesions display disrupted vascular lumens, enlarged capillary cavities, loss of proper neuro-vascular associations and an inflammatory reaction. The CCM lesions also exhibit damaged neuronal architecture, the likely cause of neurologic defects, such as ataxia and seizure. These mice represent the first CCM2 animal model for CCM and should provide the means to elucidate disease mechanisms and evaluate therapeutic strategies for human CCM.


Efferent synaptic connections of grafted dopaminergic neurons reinnervating the host neostriatum: a tyrosine hydroxylase immunocytochemical study.
In adult rats with a unilateral 6-hydroxydopamine-induced destruction of the nigrostriatal dopamine (DA) pathway, grafts of embryonic substantia nigra can establish a new dopaminergic terminal fiber plexus in the previously denervated neostriatum and compensate for some of the behavioral deficits induced by the nigrostriatal lesion. In the present study the synaptic connections of the ingrowing DA fibers from the graft were analyzed ultrastructurally, using immunocytochemical localization of tyrosine hydroxylase (TH), in animals whose lesion-induced motor asymmetry had been completely compensated for by the nigral grafts. In two of the animals, horseradish peroxidase-wheatgerm agglutinin conjugate was injected into the graft in order to trace possible reciprocal afferent connections to the graft from the host striatum. TH-immunoreactive axons from the graft were seen to make abundant symmetric synapses with neuronal elements in the host neostriatum. Between 85 and 90% of these synapses were on dendritic shafts and spines, and the rest were on neuronal perikarya. Two principal targets were identified: dendrites of spiny neurons, the majority of which are likely to be striatal projection neurons; and the cell bodies of giant neurons, most (or perhaps all) of which are known to be cholinergic interneurons. The synapses made on dendritic spines, which constituted about 40% of all TH-positive synapses formed by the TH-positive neurons in the graft, resembled those seen in normal animals, both in that they made contacts with spine necks and in that they invariably were associated with an asymmetric TH-negative synapse contacting the spine head. The innervation of the giant cell perikarya, which constituted about 6% of all TH-positive synapses found, was strikingly abnormal in that the graft-derived TH-positive fibers formed dense pericellular "baskets" selectively around the giant cell bodies. Such arrangements were never seen in the normal striatum, nor did they occur in the intact contralateral striatum in the grafted animals. It is proposed that this apparent dopaminergic hyperinnervation from the graft could provide a powerful inhibition of the cholinergic interneurons in the reinnervated host striatum, and that such an inhibitory mechanism could assist in the graft-induced functional recovery by potentiating the functional effects of DA synapses terminating on the spiny efferent neurons. This dual innervation may thus help to explain why restoration of only a small proportion of the striatal DA innervation by the graft is sufficient to induce complete compensation of, e.g., motor asymmetry in the lesioned rats.(ABSTRACT TRUNCATED AT 400 WORDS)


Nerve growth factor modulates the proliferative capacity of the intrahepatic biliary epithelium in experimental cholestasis.
We evaluated the expression of neurotrophins in rat cholangiocytes and the role and mechanisms by which nerve growth factor (NGF) modulates cholangiocyte proliferation. The expression of neurotrophins and their receptors was investigated by immunohistochemistry in liver sections and reverse-transcription polymerase chain reaction and immunoblots in isolated cholangiocytes. In vitro, the effect of NGF on cholangiocyte proliferation and signal transduction was investigated by immunoblotting for proliferating cell nuclear antigen, phosphorylated AKT (p-AKT), phosphorylated extracellular signal-regulated kinase 1/2 (p-ERK1/2), phosphorylated c-jun-N-terminal kinase, and phosphorylated p38. In vivo, rats that had undergone bile duct ligation (BDL) were treated with an anti-NGF antibody to immunoneutralize NGF and bile duct mass, proliferation, apoptosis, and inflammation were investigated by immunohistochemistry. NGF and its TrkA receptor were expressed by normal rat cholangiocytes and up-regulated following BDL. Cholangiocytes secrete NGF, and secretion is increased in proliferating BDL cholangiocytes. In vitro, NGF stimulated cholangiocyte proliferation, which was associated with enhanced p-AKT and p-ERK1/2 expression. NGF proliferation in vitro was partially blocked by the MEK inhibitor (UO126) and completely ablated by the phosphatidylinositol 3-kinase inhibitor (wortmannin). In vitro, NGF and estrogens have an additive effect on cholangiocyte proliferation by acting on phosphorylated TrkA and p-ERK1/2. In vivo, immunoneutralization of NGF decreased bile duct mass in BDL rats, which was associated with depressed proliferation and enhanced apoptosis and with increased portal inflammation. Cholangiocytes secrete NGF and express NGF receptors. NGF induces cholangiocyte proliferation by activating the ERK and, predominantly, the phosphatidylinositol 3-kinase pathway and exerts an additive effect in combination with estrogens on proliferation.


The espD full gene as a potential biomarker in active pulmonary tuberculosis.
Pulmonary tuberculosis (PTB) is still a major health problem worldwide. The espD has a potential to be a new biomarker because it is important for the espA, espC, and ESX-1 protein secretion system that are actively expressed in active multiplication of Mycobacterium tuberculosis complex. A total of 55 sputum samples and 41 culture isolates from newly diagnosed PTB patients at Dr. Soetomo Academic Hospital were collected from September 2016 to April 2019. The tested samples using polymerase chain reaction targeted 555 bp of espD gene and sequencing. Clone Manager Version 6 and NCBI BLAST were used to align the gene sequence against wild-type M. tuberculosis. The prediction of T-cell epitope in espD gene was detected by GENETYX. The three-dimensional (3D) structure of espD was modeled by SWISS-MODEL and I-TASSER and was visualized with PyMOL. From 55 sputum samples, 43 (78.18%) showed positive results, and all culture isolates showed positive results. In addition, all sequenced samples showed 100% homolog with M. tuberculosis H37Rv gene without detected variant or mutation. There were four T-cell epitopes that could be obtained. The 3D model had a I-TASSER confidence score of 3.91 with estimated RMSD of approximately 14.5 Å. The structure consists of a main fold of a three-stranded antiparallel β-sheet and a long α-helix surrounded by several minor secondary structures. This study provides a brief information about the sequence, epitope prediction, and 3D structure of EspD protein from M. tuberculosis strains in Indonesia.


Effect of Neurotoxin Exposure on Blood Biomarkers of Neurodegeneration and Alzheimer Disease.
To determine whether exposure to neurotoxins in midlife is associated with changes in blood-based biomarkers of neurodegeneration and Alzheimer disease pathology. Blood cadmium, lead, neurofilament light (NfL) chain, total tau (TTau), and amyloid beta (Aβ) 40 and Aβ42 concentrations were measured in 1516 participants in the Beaver Dam Offspring Study. Linear mixed-effect models were used to determine associations between baseline cadmium and lead levels and baseline NfL, TTau, and Aβ42/Aβ40, and 10-year change in concentrations using repeated measures of these biomarkers as the outcome. In women, higher cadmium and lead levels were associated with higher baseline TTau concentrations. A higher baseline cadmium level was associated with lower baseline Aβ42/Aβ40 in both men and women. In age-sex-adjusted models, a doubling in baseline cadmium level was associated with a 0.2% (95% CI: 0.0, 0.3) higher increase per year in NfL concentrations. In men, a doubling of baseline lead level was associated with a 0.9% (95% CI: 0.1, 1.7) higher increase per year in TTau concentration. Participants with relatively higher levels of cadmium and lead had blood biomarker concentrations consistent with more neuronal damage and Alzheimer disease pathology. Environmental exposure to neurotoxins may contribute to neurodegeneration.


CCD imaging in cryospectrophotometric determination of microvascular oxyhemoglobin saturations.
A microspectrophotometric imaging method has been developed for localized measurements of intravascular oxyhemoglobin (HbO2) saturations in microvessels from sections of quick-frozen tissue. HbO2 saturation was calculated from the absorption spectrum of red blood cells measured at five selected wavelengths in the 520- to 570-nm range. We combined the use of narrow-bandwidth interference filters and a CCD camera mounted on a microscope to obtain one gray image of the sample at each wavelength. Each pixel is a quantitative measure of transmitted light intensity from the tissue sample at that location. A linear calibration curve for blood frozen in vitro (humans and mice) and in vivo (mice) was obtained using a multicomponent analysis. Oxy- and deoxyhemoglobin were assumed to be the only hemoglobin components present. A constant term compensates for light loss due to scattering on red blood cells and ice crystals. The standard error in single measurements of HbO2 saturation was 5%. The present method allows off-line analysis of the HbO2 saturation distribution within a microvessel network and offers new possibilities for comparative morphological studies.


Gram-negative outer membrane vesicles: beyond the cell surface.
Considerable interest has recently mounted regarding the biological roles of Gram-negative outer membrane vesicles (MVs). The first discovery of MVs was made over four decades ago, and it is now clear that most Gram-negative bacteria produce MVs, with Pseudomonas aeruginosa and Escherichia coli as the most extensively studied. Much of our knowledge of the biological roles of MVs and mechanism of MV formation is due to T.J. Beveridge and colleagues. Beveridge pioneered the field of MV research not only by enhancing our understanding of MV function, but also through the application of a wide variety of physical, chemical, and genetic techniques to complement his elegant electron microscopy investigations. Here we review the contributions of Beveridge's group to our understanding of MV biology.


Effects of nitroprusside on the bradykinin responsiveness of human fibroblasts.
The effects of agents that cause vasodilatation and hypotension, such as endogenously produced bradykinin (BK) or the drug nitroprusside (NP), appear to result from effects on cyclic nucleotides (cGMP, cAMP) and arachidonate metabolism. Cultured human fibroblasts, which possess B2 BK receptors and respond to NP with an increase in cGMP, were used to study the interaction of these agents at the molecular level. Addition of BK or NP to cultured human fibroblasts caused a rapid increase in cGMP. The effect of NP was usually maximal within 30 sec, after which cGMP content declined. The increase in cGMP produced by BK reached a maximum at approximately 1 min and then fell; the rise with NP was more than 10 times that with BK. At 30 sec, cGMP content with NP plus BK was less than with NP alone. At later times, however, effects of BK and NP were slightly more than additive and maximal cGMP levels were reached at 90 sec. BK increased prostaglandin production by the fibroblasts; it is believed that the kinin-induced elevation in cAMP content is secondary to increased prostaglandin formation. NP caused a small, early increase in cAMP without significant effect on prostaglandin I2 (PGI2); after 2.5 min, effects on PGI2 and cAMP were greater with BK and NP than with BK alone. To study further the roles of arachidonate metabolites in the fibroblast response to BK and NP, the cyclooxygenase inhibitor, indomethacin, and the combined lipoxygenase and cyclooxygenase inhibitor, 5,8,11,14-eicosatetraynoic acid (ETYA), were added to fibroblasts prior to BK or NP. Increases in cAMP or PGI2 with BK or BK plus NP were blocked by indomethacin or ETYA. These effects of BK or BK plus NP on cAMP thus appear to be mediated through cyclooxygenase products of arachidonate metabolism. Indomethacin and ETYA did not affect cGMP in the presence of BK plus NP but enhanced NP-stimulated cGMP accumulation by 40-50%; effects of NP on cGMP may be independent of or perhaps inhibited by cyclooxygenase derivatives. Cellular responses to BK plus NP differed quantitatively and temporally from the sum of effects of BK and NP alone. Through interactions of this type, in vivo responses to drugs like NP may be influenced by levels of BK or similar endogenous mediators.


Professional practice in contested territory: child health nurses and maternal sadness.
This research explored the extent to which Child and Family Health Nurses (CHNs) exercise autonomy as specialist health professionals within community health settings. Using postnatal depression (PND) as an example we examined if, and how, CHNs exercised autonomy within their practice, drawing on their specialised body of knowledge. A qualitative study was undertaken using in-depth interviews with 10 CHNs practising in Tasmania, Australia. We analysed the interview data thematically to identify how CHNs defined and discussed PND, the strategies they drew on in their practice, and indicators of professional identity and autonomy in their descriptions of practice as it pertained to PND. Three themes emerged from this analysis: alternative understandings of maternal sadness; technicality, indeterminancy and tacit knowledge; and identification of professional boundaries. We found that CHNs' autonomous practice was informed by understandings distinct from dominant perspectives, suggesting that this specialist area of nursing practice exhibits extensive indicators of professionalism that have been largely unrecognised. This has implications for both CHNs' professional identity and the provision of services available to new mothers and their babies.


Health locus of control in a pandemic situation: cross-cultural differences between European and Asian respondents.
The present cross-cultural study examined the health locus of control construct during the COVID-19 pandemic. The scientific purpose of the study was to determine whether, during the pandemic situation, cultural and sex differences influence the health locus of control construct and change the internal health locus of control (IHLC), powerful others health locus of control (PHLC), and chance health locus of control (CHLC). A total of 2617 recipients aged 18-70 years from Asia (China, India, and Indonesia), and Europe (Bulgaria, Germany, and Hungary) completed a questionnaire about their health. The participants completed an online version of the Multidimensional Health Locus of Control Scale - Form A. The survey shows that in a pandemic life-threatening situation, most individuals strive to rely on IHLC and/or PHLC, and fewer of them tend to rely on CHLC. However, there are differences (p < .001) between the two cultural samples: the representatives of Asian collectivistic culture are more dominated by PHLC, compared to the representatives of the European individualistic culture. When the comparison is between individuals from different cultures, sex differentiation affects the health locus of control, and as a result, significant differences in relation to IHLC, PHLC, and CHLC levels (p < .05) appear. In conclusion, the study indicates that cultural differences influence both the IHLC and PHLC levels, and that Asian participants are dominated by PHLC more than European respondents. Asian females are more likely to seek support from powerful others (PHLC) compared to European women, who perceive themselves as more independent. Asian male participants are prepared to rely on powerful others (doctors or medical institutions), while European male respondents are prone to rely on themselves mainly (IHLC). The results show that sex differences do not significantly affect the health locus of control within the same cultural group.


Assessing the effectiveness of house-to-house visits on routine oral polio immunization completion and tracking of defaulters.
Strengthening routine immunization is one of the four prongs of the Global Polio Eradication Initiative. Using data collected through 30-cluster sample household surveys of caretakers of children aged 12-23 months, this paper assessed the effectiveness of house-to-house visits on routine oral polio immunization completion, using simple frequency tables, bivariate and multivariate logistic regression analyses. Logistic regression results demonstrated that children in households where the caregivers reported receiving a household visit by health workers were more likely to be fully immunized for polio through routine immunization than other children, although results were significant only after correcting for confounders. In Ethiopia and India, children of caregivers who remembered a house-to-house visit were significantly and positively associated with routine polio vaccination completion (OR = 2.2 and OR = 2.2 respectively). In Angola, the association was positive, though not significant (OR = 1.3). The evidence suggests that targeting high-risk areas for house-to-house visits played a role in increasing routine polio vaccination.


Dissecting the dynamic changes of 5-hydroxymethylcytosine in T-cell development and differentiation.
The discovery of Ten Eleven Translocation proteins, enzymes that oxidize 5-methylcytosine (5mC) in DNA, has revealed novel mechanisms for the regulation of DNA methylation. We have mapped 5-hydroxymethylcytosine (5hmC) at different stages of T-cell development in the thymus and T-cell differentiation in the periphery. We show that 5hmC is enriched in the gene body of highly expressed genes at all developmental stages and that its presence correlates positively with gene expression. Further emphasizing the connection with gene expression, we find that 5hmC is enriched in active thymus-specific enhancers and that genes encoding key transcriptional regulators display high intragenic 5hmC levels in precursor cells at those developmental stages where they exert a positive effect. Our data constitute a valuable resource that will facilitate detailed analysis of the role of 5hmC in T-cell development and differentiation.


Microbial domestication signatures of Lactococcus lactis can be reproduced by experimental evolution.
Experimental evolution is a powerful approach to unravel how selective forces shape microbial genotypes and phenotypes. To this date, the available examples focus on the adaptation to conditions specific to the laboratory. The lactic acid bacterium Lactococcus lactis naturally occurs on plants and in dairy environments, and it is proposed that dairy strains originate from the plant niche. Here we investigate the adaptation of a L. lactis strain isolated from a plant to a dairy niche by propagating it for 1000 generations in milk. Two out of three independently evolved strains displayed significantly increased acidification rates and biomass yields in milk. Genome resequencing, revealed six, seven, and 28 mutations in the three strains, including point mutations in loci related to amino acid biosynthesis and transport and in the gene encoding MutL, which is involved in DNA mismatch repair. Two strains lost a conjugative transposon containing genes important in the plant niche but dispensable in milk. A plasmid carrying an extracellular protease was introduced by transformation. Although improving growth rate and growth yield significantly, the plasmid was rapidly lost. Comparative transcriptome and phenotypic analyses confirmed that major physiological changes associated with improved growth in milk relate to nitrogen metabolism and the loss or down-regulation of several pathways involved in the utilization of complex plant polymers. Reproducing the transition from the plant to the dairy niche through experimental evolution revealed several genome, transcriptome, and phenotype signatures that resemble those seen in strains isolated from either niche.


Revised form of the Belief in Good Luck Scale in a Turkish sample.
The aim of the study was to explore the patterns of belief in luck in a Turkish sample. For this purpose, a shorter version of Freedman and Darke's Belief in Good Luck Scale was given, including some more items related to the beliefs in 'consistent' versus 'variant' patterns of luck. This scale, together with Turkish versions of Rosenberg's measure of Global Self-esteem, Rotter's Internal-External Locus of Control Scale, and the Satisfaction With Life Scale of Diener, Emmons, Larsen, and Griffin, was administered to a group of 173 (96 men, 77 women) undergraduate students from the Middle East Technical University attending an elective general psychology course. The mean age was 21 yr. (SD=2.1). The factor composition of the extended scale consisted of three scales, similar to the original version. Loading of new items on the General Belief in Luck factor indicates that this factor may describe a belief in an organised pattern of luck.


Performance of whole-body integrated 18F-FDG PET/MR in comparison to PET/CT for evaluation of malignant bone lesions.
Because of its higher soft-tissue contrast, whole-body integrated PET/MR offers potential advantages over PET/CT for evaluation of bone lesions. However, unlike PET/CT, PET/MR ignores the contribution of cortical bone in the attenuation map. Thus, the aims of this study were to evaluate the diagnostic performance of whole-body integrated (18)F-FDG PET/MR specifically for bone lesions and to analyze differences in standardized uptake value (SUV) quantification between PET/MR and PET/CT. One hundred nineteen patients with (18)F-FDG-avid primary malignancies underwent a single-injection, dual-imaging protocol using (18)F-FDG on a PET/CT scanner and a subsequent PET/MR scan with a T1-weighted volumetric interpolated breath-hold examination (VIBE) Dixon sequence for attenuation correction and an unenhanced coronal T1-weighted turbo spin-echo (TSE) sequence for bone analysis. Three sets of images (CT with PET [from PET/CT; set A], T1-weighted VIBE Dixon with PET [set B], and T1-weighted TSE with PET [both from PET/MR; set C]) were analyzed. Two readers rated every lesion using a 4-point scale for lesion conspicuity on PET, a 4-point scale for anatomic allocation of PET-positive lesions, and a 5-point scale for the nature of every lesion based on its appearance on morphologic imaging and uptake on PET. For all lesions and for representative regions of normal bone, SUV analysis was performed for PET/MR and PET/CT. In total, 98 bone lesions were identified in 33 of 119 patients, and 630 regions of normal bone were analyzed. Visual lesion conspicuity on PET was comparable for PET/CT (mean rating, 2.82 ± 0.45) and PET/MR (2.75 ± 0.51; P = 0.3095). Anatomic delineation and allocation of suggestive lesions was significantly superior with T1-weighted TSE MRI (mean rating, 2.84 ± 0.42) compared with CT (2.57 ± 0.54, P = 0.0001) or T1-weighted VIBE Dixon MRI (2.57 ± 0.54, P = 0.0002). No significant difference in correct classification of malignant bone lesions was found among sets A (85/90), B (84/90), and C (86/90). For bone lesions and regions of normal bone, a highly significant correlation existed between the mean SUVs for PET/MR and PET/CT (R = 0.950 and 0.917, respectively, each P < 0.001). However, substantially lower mean SUVs were found for PET/MR than for PET/CT both for bone lesions (12.4% ± 15.5%) and for regions of normal bone (30.1% ± 27.5%). Compared with PET/CT, fully integrated whole-body (18)F-FDG PET/MR is technically and clinically robust for evaluation of bone lesions despite differences in attenuation correction. PET/MR, including diagnostic T1-weighted TSE sequences, was superior to PET/CT for anatomic delineation and allocation of bone lesions. This finding might be of clinical relevance in selected cases--for example, primary bone tumors, early bone marrow infiltration, and tumors with low uptake on PET. Thus, a diagnostic T1-weighted TSE sequence is recommended as a routine protocol for oncologic PET/MR.


Multicenter Noninferiority Evaluation of Hain GenoType MTBDRplus Version 2 and Nipro NTM+MDRTB Line Probe Assays for Detection of Rifampin and Isoniazid Resistance.
Less than 30% of multidrug-resistant tuberculosis (MDR-TB) patients are currently diagnosed, due to laboratory constraints. Molecular diagnostics enable rapid and simplified diagnosis. Newer-version line probe assays have not been evaluated against the WHO-endorsed Hain GenoType MTBDRplus (referred to as Hain version 1 [V1]) for the rapid detection of rifampin (RIF) and isoniazid (INH) resistance. A two-phase noninferiority study was conducted in two supranational reference laboratories to allow head-to-head comparisons of two new tests, Hain Genotype MTBDRplus version 2 (referred to as Hain version 2 [V2]) and Nipro NTM+MDRTB detection kit 2 (referred to as Nipro), to Hain V1. In phase 1, the results for 379 test strains were compared to a composite reference standard that used phenotypic drug susceptibility testing (DST) and targeted sequencing. In phase 2, the results for 644 sputum samples were compared to a phenotypic DST reference standard alone. Using a challenging set of strains in phase 1, the values for sensitivity and specificity for Hain V1, Hain V2, and Nipro, respectively, were 90.3%/98.5%, 90.3%/98.5%, and 92.0%/98.5% for RIF resistance detection and 89.1%/99.4%, 89.1%/99.4%, and 89.6%/100.0% for INH resistance detection. Testing of sputa in phase 2 yielded values for sensitivity and specificity of 97.1%/97.1%, 98.2%/97.8%, and 96.5%/97.5% for RIF and 94.4%/96.4%, 95.4%/98.8%, and 94.9%/97.6% for INH. Overall, the rates of indeterminate results were low, but there was a higher rate of indeterminate results with Nipro than with Hain V1 and V2 in samples with low smear grades. Noninferiority of Hain V2 and Nipro to Hain V1 was demonstrated for RIF and INH resistance detection in isolates and sputum specimens. These results serve as evidence for WHO policy recommendations on the use of line probe assays, including the Hain V2 and Nipro assays, for MDR-TB detection.