National Bone Health Alliance Bone Turnover Marker Project: current practices and the need for US harmonization, standardization, and common reference ranges.
This position paper reviews how the National Bone Health Alliance (NBHA) will execute a project to help assure health professionals of the clinical utility of bone turnover markers; the current clinical approaches concerning osteoporosis and the status and use of bone turnover markers in the USA; the rationale for focusing this effort around two specific bone turnover markers; the need to standardize bone marker sample collection procedures, reference ranges, and bone turnover marker assays in clinical laboratories; and the importance of harmonization for future research of bone turnover markers. Osteoporosis is a major global health problem, with the prevalence and incidence of osteoporosis for at-risk populations estimated to be 44 million Americans. The potential of bone markers as an additional tool for health care professionals to improve patient outcomes and impact morbidity and mortality is crucial in providing better health care and addressing rising health care costs. This need to advance the field of bone turnover markers has been recognized by a number of organizations, including the International Osteoporosis Foundation (IOF), National Osteoporosis Foundation, International Federation of Clinical Chemistry, and Laboratory Medicine (IFCC), and the NBHA. This position paper elucidates how this project will standardize bone turnover marker sample collection procedures in the USA, establish a USA reference range for one bone formation (serum procollagen type I N propeptide, s-PINP) and one bone resorption (serum C-terminal telopeptide of type I collagen, s-CTX) marker, and standardize bone turnover marker assays used in clinical laboratories. This effort will allow clinicians from the USA to have confidence in their use of bone turnover markers to help monitor osteoporosis treatment and assess future fracture risk. This project builds on the recommendations of the IOF/IFCC Bone Marker Standards Working Group by developing USA reference standards for s-PINP and s-CTX, the markers identified as most promising for use as reference markers. The goals of this project will be realized through the NBHA and will include its governmental, academic, for-profit, and non-profit sector stakeholders as well as major academic and commercial laboratories. Upon completion, a parallel effort will be pursued to make bone turnover marker measurements reliable and accepted by all health care professionals for facilitating treatment decisions and ultimately be reimbursed by all health insurance payers. Successful completion of this project will help assure health professionals from the USA of the clinical utility of bone turnover markers and ties in with the parallel effort of the IOF/IFCC to develop worldwide bone turnover reference ranges.


Patterns of distress in US medical students.
How multiple forms of psychological distress coexist in individual medical students has not been formally studied. To explore the prevalence of various forms of distress in medical students and their relationship to recent suicidal ideation or serious thoughts of dropping out of school. All medical students at seven US schools were surveyed with standardized instruments to evaluate burnout, depression, stress, mental quality of life (QOL), physical QOL, and fatigue. Additional items explored recent suicidal ideation and serious thoughts of dropping out of medical school. Nearly all (1846/2246, 82%) of medical students had at least one form of distress with 1066 (58%) having ≥3 forms of distress. A dose-response relationship was found between the number of manifestations of distress and recent suicidal ideation or serious thoughts of dropping out. For example, students with 2, 4, or 6 forms of distress were 5, 15, and 24 fold, respectively, more likely to have suicidal ideation than students with no forms of distress assessed. All forms of distress were independently associated with suicidal ideation or serious thoughts of dropping out on multivariable analysis. Most medical students experience ≥1 manifestation of distress with many experiencing multiple forms of distress simultaneously. The more forms of distress experienced the greater the risk for suicidal ideation and thoughts of dropping out of medical school.


Hormesis and adaptive cellular control systems.
Hormetic dose response occurs for many endpoints associated with exposures of biological organisms to environmental stressors. Cell-based U- or inverted U-shaped responses may derive from common processes involved in activation of adaptive responses required to protect cells from stressful environments. These adaptive pathways extend the region of cellular homeostasis and are protective against ultimate cell, organ, and system toxicity. However, the activation of stress responses carries a significant energetic cost to the cell, leading to alterations of a variety of basal cellular functions in adapted or stressed cells. This tradeoff of resources between the unstressed and adapted states may lead to U-or inverted U-shaped dose response curves for some precursor endpoints. We examine this general hypothesis with chlorine, a prototype oxidative stressor, using a combination of cellular studies with gene expression analysis of response pathways and with computational modeling of activation of control networks. Discrete cellular states are expected as a function of exposure concentration and duration. These cellular states include normal functioning state, adaptive and stressed states at mild to intermediate exposures, and overt toxicity in the presence of an overwhelming concentration of stressors. These transitions can be used to refine default risk assessment practices that do not currently accommodate adaptive responses.


The global tree restoration potential.
The restoration of trees remains among the most effective strategies for climate change mitigation. We mapped the global potential tree coverage to show that 4.4 billion hectares of canopy cover could exist under the current climate. Excluding existing trees and agricultural and urban areas, we found that there is room for an extra 0.9 billion hectares of canopy cover, which could store 205 gigatonnes of carbon in areas that would naturally support woodlands and forests. This highlights global tree restoration as our most effective climate change solution to date. However, climate change will alter this potential tree coverage. We estimate that if we cannot deviate from the current trajectory, the global potential canopy cover may shrink by ~223 million hectares by 2050, with the vast majority of losses occurring in the tropics. Our results highlight the opportunity of climate change mitigation through global tree restoration but also the urgent need for action.


Investigation of the mechanism of beta 2-agonist-induced activation of the renin-angiotensin system.
1. We have previously described activation of the renin-angiotensin system in asthma, and also by high-dose nebulized beta 2-agonists. In this study we sought to determine the mechanism responsible. 2. The influence of the angiotensin-converting enzyme inhibitor, lisinopril, on the response of the renin-angiotensin system and serum potassium to nebulized salbutamol was investigated in a randomized, double-blind, crossover study in eight healthy volunteers using a factorial block design. On study days, subjects received lisinopril 20 mg orally or identical placebo tablets followed 3 h later by nebulized salbutamol or placebo inhalation; plasma renin, angiotensin II, serum angiotensin-converting enzyme and potassium were measured at intervals for 120 min after inhalation. 3. Following salbutamol, plasma renin and angiotensin II concentrations were increased significantly compared with placebo [mean (SEM) plasma renin of 61.7 (15.6) mu-units/ml and angiotensin II of 17.7 (5.4) pg/mol 15 min after salbutamol, P < 0.05 versus placebo]. Baseline plasma renin concentrations were increased [160.1 (20.6) mu-units/ml] and baseline plasma angiotensin II concentrations were reduced [1.4 (0.1) pg/ml] by lisinopril, P < 0.05 versus placebo in each case. Inhibition of angiotensin-converting enzyme completely inhibited this salbutamol-induced rise in plasma angiotensin II [mean (SEM) plasma angiotensin II of 1.5 (0.4) pg/ml 15 min after salbutamol, P < 0.05 versus placebo] but had no effect on the changes in plasma renin concentrations after the beta 2-agonist [mean (SEM) plasma renin of 198.4 (18.9) mu-units/ml 15 min after salbutamol].(ABSTRACT TRUNCATED AT 250 WORDS)


Bacterial and protozoal agents of feline vector-borne diseases in domestic and stray cats from southern Portugal.
Feline vector-borne diseases (FVBD) have emerged in recent years, showing a wider geographic distribution and increased global prevalence. In addition to their veterinary importance, domestic cats play a central role in the transmission cycles of some FVBD agents by acting as reservoirs and sentinels, a circumstance that requires a One Health approach. The aim of the present work was to molecularly detect feline vector-borne bacteria and protozoa with veterinary and zoonotic importance, and to assess associated risk factors in cats from southern Portugal. Six hundred and forty-nine cats (320 domestic and 329 stray), from veterinary medical centres and animal shelters in southern Portugal, were studied. Anaplasma spp./Ehrlichia spp., Babesia spp., Bartonella spp., Borrelia burgdorferi sensu lato, Hepatozoon spp. and Leishmania spp. infections were evaluated by polymerase chain reaction (PCR) in blood samples. One hundred and ninety-four (29.9%) cats were PCR-positive to at least one of the tested genera or complex of FVBD agents. Sixty-four (9.9%) cats were positive to Leishmania spp., 56 (8.6%) to Hepatozoon spp., 43 (6.6%) to Babesia spp., 35 (5.4%) to Anaplasma spp./Ehrlichia spp., 19 (2.9%) to Bartonella spp. and 14 (2.2%) to B. burgdorferi s.l. Thirty-three (5.1%) cats were positive to two (n = 29) or three (n = 4) genera/complex. Babesia vogeli, Bartonella clarridgeiae, Bartonella henselae, Ehrlichia canis, Hepatozoon felis and Leishmania infantum were identified by DNA sequencing. The occurrence of FVBD agents in southern Portugal, some of them with zoonotic character, emphasizes the need to alert the veterinary community, owners and public health authorities for the risk of infection. Control measures should be implemented to prevent the infection of cats, other vertebrate hosts and people.


The Roles and Mechanisms of Actions of Vitamin C in Bone: New Developments.
Vitamin C is an important antioxidant and cofactor that is involved in the regulation of development, function, and maintenance of several cell types in the body. Deficiencies in vitamin C can lead to conditions such as scurvy, which, among other ailments, causes gingivia, bone pain, and impaired wound healing. This review examines the functional importance of vitamin C as it relates to the development and maintenance of bone tissues. Analysis of several epidemiological studies and genetic mouse models regarding the effect of vitamin C shows a positive effect on bone health. Overall, vitamin C exerts a positive effect on trabecular bone formation by influencing expression of bone matrix genes in osteoblasts. Recent studies on the molecular pathway for vitamin C actions that include direct effects of vitamin C on transcriptional regulation of target genes by influencing the activity of transcription factors and by epigenetic modification of key genes involved in skeletal development and maintenance are discussed. With an understanding of mechanisms involved in the uptake and metabolism of vitamin C and knowledge of precise molecular pathways for vitamin C actions in bone cells, it is possible that novel therapeutic strategies can be developed or existing therapies can be modified for the treatment of osteoporotic fractures.


Recombinant protein production from stable mammalian cell lines and pools.
We highlight recent developments for the production of recombinant proteins from suspension-adapted mammalian cell lines. We discuss the generation of stable cell lines using transposons and lentivirus vectors (non-targeted transgene integration) and site-specific recombinases (targeted transgene integration). Each of these methods results in the generation of cell lines with protein yields that are generally superior to those achievable through classical plasmid transfection that depends on the integration of the transfected DNA by non-homologous DNA end-joining. This is the main reason why these techniques can also be used for the generation of stable cell pools, heterogenous populations of recombinant cells generated by gene delivery and genetic selection without resorting to single cell cloning. This allows the time line from gene transfer to protein production to be reduced.


Work schedules of long-distance truck drivers before and after 2004 hours-of-service rule change.
Federal rules regulate work hours of interstate commercial truck drivers. On January 4, 2004, a new work rule was implemented, increasing daily and weekly maximum driving limits and daily off-duty requirements. The present study assessed changes in long-distance truck drivers' reported work schedules and reported fatigued driving after the rule change. Associations between reported rule violations, fatigued driving, and schedule as well as other characteristics were examined. Samples of long-distance truck drivers were interviewed face-to-face in two states immediately before the rule change (November-December 2003) and about 1 year (November-December 2004) and 2 years (November-December 2005) after the change. Drivers reported substantially more hours of driving after the rule change. Most drivers reported regularly using a new restart provision, which permits a substantial increase in weekly driving. Reported daily off-duty and sleep time increased. Reported incidents of falling asleep at the wheel of the truck increased between 2003 (before the rule change) and 2004 and 2005 (after the change); in 2005 about one fifth of drivers reported falling asleep at the wheel in the past month. The frequency of reported rule violations under the old and new rules was similar. The percentage of trucks with electronic on-board recorders increased significantly to almost half the fleet; only a few drivers were using automated recorders to report rule compliance. More than half of drivers said that requiring automated recorders on all large trucks to enforce driving-hour limits would improve compliance with work rules. Based on the 2004-2005 survey data, drivers who reported more frequent rule violations were significantly more likely to report fatigued driving. Predictors of reported violations included having unrealistic delivery schedules, longer wait times to drop off or pick up loads, difficulty finding a legal place to stop or rest, and driving a refrigerated trailer. Reported truck driver fatigue increased after the new rule was implemented, suggesting that the rule change may not have achieved the goal of reducing fatigued driving. Reported violations of the work rules remain common. Because many trucks already have electronic recorders, requiring them as a means of monitoring driving hours appears feasible.


Development and validation of a microRNA-based signature (MiROvaR) to predict early relapse or progression of epithelial ovarian cancer: a cohort study.
Risk of relapse or progression remains high in the treatment of most patients with epithelial ovarian cancer, and development of a molecular predictor could be a valuable tool for stratification of patients by risk. We aimed to develop a microRNA (miRNA)-based molecular classifier that can predict risk of progression or relapse in patients with epithelial ovarian cancer. We analysed miRNA expression profiles in three cohorts of samples collected at diagnosis. We used 179 samples from a Multicenter Italian Trial in Ovarian cancer trial (cohort OC179) to develop the model and 263 samples from two cancer centres (cohort OC263) and 452 samples from The Cancer Genome Atlas epithelial ovarian cancer series (cohort OC452) to validate the model. The primary clinical endpoint was progression-free survival, and we adapted a semi-supervised prediction method to the miRNA expression profile of OC179 to identify miRNAs that predict risk of progression. We assessed the independent prognostic role of the model using multivariable analysis with a Cox regression model. We identified 35 miRNAs that predicted risk of progression or relapse and used them to create a prognostic model, the 35-miRNA-based predictor of Risk of Ovarian Cancer Relapse or progression (MiROvaR). MiROvaR was able to classify patients in OC179 into a high-risk group (89 patients; median progression-free survival 18 months [95% CI 15-22]) and a low-risk group (90 patients; median progression-free survival 38 months [24-not estimable]; hazard ratio [HR] 1·85 [1·29-2·64], p=0·00082). MiROvaR was a significant predictor of progression in the two validation sets (OC263 HR 3·16, 95% CI 2·33-4·29, p<0·0001; OC452 HR 1·39, 95% CI 1·11-1·74, p=0·0047) and maintained its independent prognostic effect when adjusted for relevant clinical covariates using multivariable analyses (OC179: adjusted HR 1·48, 95% CI 1·03-2·13, p=0·036; OC263: adjusted HR 3·09 [2·24-4·28], p<0·0001; and OC452: HR 1·41 [1·11-1·79], p=0·0047). MiROvaR is a potential predictor of epithelial ovarian cancer progression and has prognostic value independent of relevant clinical covariates. MiROvaR warrants further investigation for the development of a clinical-grade prognostic assay. AIRC and CARIPLO Foundation.


[State of dentition and its impact on the capacity of elders to perform daily activities].
The aim of this study was to evaluate the effect of dental state on the capacity of elderly individuals to carry out their daily activities during the previous six months. A transversal study was conducted in the Mexican Institute of Social Security (IMSS) with insured elderly persons residing in southwestern Mexico City. Home interviews were conducted to gather sociodemographic data, data on the use of oral health services, problems in carrying out daily activities, and their relationship with the state of dentition over the past six months, utilizing the Oral Impacts on Daily Performance (OIDP) index. Later, three oral health professionals trained in determining the state of dentition according to World Health Organization (WHO) criteria performed a clinical examination on each elderly subject. A total of 540 elderly individuals were studied. The prevalence of elderly persons with problems in carrying out their daily activities was 21.5%. The main problems in daily activities were eating (14.4%), speaking (8.7%), not becoming irritated (5.4%), brushing their teeth(5%), and inability to enjoy contact with people (4.4%). There is a positive OIDP correlation between number of teeth lost and the decayed, missing, filled teeth(DMFT) index (p<0.05) and a negative correlation with the number of functional and filled teeth. The state of dentition in the elderly has repercussions on the capacity to carry out daily activities, with quality of life most seriously affected with regard to eating and enjoying food, as well as with respect to correct pronunciation.


Functional connectivity of the orbitofrontal cortex, anterior cingulate cortex, and inferior frontal gyrus in humans.
Parcellation of the orbitofrontal cortex, anterior cingulate cortex, and inferior frontal gyrus based on their functional connectivity with the whole brain in resting state fMRI with 654 participants was performed to investigate how these regions with different functions in reward, emotion and their disorders are functionally connected to each other and to the whole brain. The human medial and lateral orbitofrontal cortex, the ventromedial prefrontal cortex, the anterior cingulate cortex, and the right and left inferior frontal gyrus have different functional connectivity with other brain areas and with each other; and each of these regions has several parcels with different functional connectivity with other brain areas. In terms of functional connectivity, the lateral orbitofrontal cortex extends especially on the right into the orbital part of the inferior frontal gyrus and provides connectivity with premotor cortical areas. The orbitofrontal cortex, especially the lateral orbitofrontal cortex, has connectivity not only with language-related areas in the inferior frontal gyrus (Broca's area), but also with the angular and supramarginal gyri. In this context, whereas the connectivity of the orbitofrontal cortex, ventromedial prefrontal cortex, and anterior cingulate cortex is symmetrical, the connectivity of the inferior frontal gyrus triangular and opercular parts is asymmetrical for the right and the left hemispheres. These findings have implications for understanding the neural bases of human emotion and decision-making, and for their disorders including depression.


How Well Do They Convert? Trending ASAPS Presentations to Publication From 1995-2010.
The American Society for Aesthetic Plastic Surgery (ASAPS) sponsors an annual conference that promotes education, advocacy, and care. There, researchers deliver abstracts as podium and poster presentations. Subsequently, ASAPS encourages submitting these research findings for publication. Yet, many never become published manuscripts. To quantify the conversion rates of oral abstract presentations to publication from 1995 to 2010. Secondary objectives included evaluating trends in presentations, publications, time to publication, and published journal distribution. Comprehensive literature search in PubMed cross-referencing oral abstract presentations and determining peer-reviewed publication status. The conversion rate and time to publication was calculated. A total of 569 oral presentations met the inclusion criteria. The mean annual presentations was 35.6. A total of 360 presentations became journal publications. The mean annual publications was 22.5. The mean conversion rate was 63.3% (R2, 0.1271; P-value of .23). The mean time to publication was 19.8 months. Most publications occurred within two years of presentation (87.5%). Publications appeared in Plastic and Reconstructive Surgery (PRS, 48.6%), Aesthetic Surgery Journal (ASJ, 27.8%), Aesthetic Plastic Surgery (APS, 5.6%), Annals of Plastic Surgery (AnnPS, 4.2%), Clinics in Plastic Surgery (CPS, 3.9%), and other journals (10%). Trending ASJ publications vs other journals in five year intervals demonstrated an increase from 18.7% to 58.8%. While the number of presentations and publications declined, the time to publication, and conversion rate remained largely the same. Despite its short existence, ASJ became the predominant journal publishing ASAPS abstracts by the end of the study period.


Impaired neutrophil function in 24p3 null mice contributes to enhanced susceptibility to bacterial infections.
Lipocalin 24p3 (24p3) is a neutrophil secondary granule protein. 24p3 is also a siderocalin, which binds several bacterial siderophores. It was therefore proposed that synthesis and secretion of 24p3 by stimulated macrophages or release of 24p3 upon neutrophil degranulation sequesters iron-laden siderophores to attenuate bacterial growth. Accordingly, 24p3-deficient mice are susceptible to bacterial pathogens for which siderophores would normally be chelated by 24p3. Specific granule deficiency (SGD) is a rare congenital disorder characterized by complete absence of proteins in secondary granules. Neutrophils from SGD patients, who are prone to bacterial infections, lack normal functions, but the potential role of 24p3 in neutrophil dysfunction in SGD is not known. In this study, we show that neutrophils from mice genetically deficient for lipocalin 24p3 (24p3(-/-)) are defective in many neutrophil functions. Specifically, neutrophils in 24p3(-/-) mice do not extravasate to sites of infection and are defective for chemotaxis. A transcriptome analysis revealed that genes that control cytoskeletal reorganization are selectively suppressed in 24p3(-/-) neutrophils. Additionally, small regulatory RNAs (microRNAs) that control upstream regulators of cytoskeletal proteins are also increased in 24p3(-/-) neutrophils. Further, 24p3(-/-) neutrophils failed to phagocytose bacteria, which may account for the enhanced sensitivity of 24p3(-/-) mice to both intracellular (Listeria monocytogenes) and extracellular (Candida albicans and Staphylococcus aureus) pathogens. Listeria does not secrete siderophores, and additionally, the siderophore secreted by Candida is not sequestered by 24p3. Therefore, the heightened sensitivity of 24p3(-/-) mice to these pathogens is not due to sequestration of siderophores limiting iron availability, but is a consequence of impaired neutrophil function.


The impact of neurosurgical procedure on cognitive resources: Results of bypass training.
Neurosurgeons are exposed to unavoidable distractions in their natural operating environment. Distractions can affect both the surgeon's concentration and the safety and duration of the surgery. Such distraction can be studied by applying a simultaneous cognitive task during a surgical procedure. We used a previously described cognitive task: a forward (DF) and backward digit (DB) repetition task to interfere with the surgeon's attention during a training bypass. A pilot study was performed to find suitable digit repetition lengths. For the main experiment, we used four-digit strings. The test task was alternated across two consecutive sutures (n = 153, 8 bypasses), followed by two consecutive control sutures without digit repetition. The duration and the number of correct answers for the digit repetition task were compared to a baseline digit repetition without simultaneous surgery. During the bypass surgery, digit repetitions (especially DB) became slower (P < 0.0001). More errors were made during DB compared to DF only during simultaneous bypass (P < 0.0001). However, we found no effect of digit repetition tasks on individual suture times (P = 0.823). The ability to engage in simultaneous tasks while performing surgery is diminished. A surgeon with extensive training can withstand external distraction without an effect on performance; however, this is achieved by partially ignoring the simultaneous task. Our data support that during surgery other cognitive tasks should be avoided to ensure safety.


Disruption of the gene encoding the ChiB1 chitinase of Aspergillus fumigatus and characterization of a recombinant gene product.
The gene encoding a major, inducible 45 kDa chitinase of Aspergillus fumigatus was cloned and analysis of the deduced amino acid sequence identified a chitinase of the fungal/bacterial class which was designated ChiB1. Recombinant ChiB1, expressed in Pichia pastoris, was shown to function by a retaining mechanism of action. That is, the beta-conformation of the chitin substrate linkage was preserved in the product in a manner typical of family 18 chitinases. Cleavage patterns with the N-acetylglucosamine (GlcNAc) oligosaccharide substrates GlcNAc(4), GlcNAc(5) and GlcNAc(6) indicated that the predominant reaction involved hydrolysis of GlcNAc(2) from the non-reducing end of each substrate. Products of transglycosylation were also identified in each incubation. Following disruption of chiB1 by gene replacement, growth and morphology of disruptants and of the wild-type strain were essentially identical. However, during the autolytic phase of batch cultures the level of chitinase activity in culture filtrate from a disruptant was much lower than the activity from the wild-type. The search for chitinases with morphogenetic roles in filamentous fungi should perhaps focus on chitinases of the fungal/plant class although such an investigation will be complicated by the identification of at least 11 putative active site domains for family 18 chitinases in the A. fumigatus TIGR database (http://www.tigr.org/).


Differential effects of nifedipine, nicorandil and nitroglycerin on the pressor responses elicited by selective alpha-1 and alpha-adrenoceptor agonists in conscious dogs.
The influence of vasodilators with varying mechanisms of action on pressor responses mediated by alpha-1 and alpha-2 adrenoceptors was investigated in chronically instrumented, conscious dogs. After ganglionic, cholinergic and beta adrenergic blockade, equipressor doses of phenylephrine (0.6 microgram/kg i.v.), a selective alpha-1 adrenoceptor agonist, and B-HT 933 (20 micrograms/kg i.v.) a selective alpha-2 adrenoceptor agonist, were administered before and in the presence of infusions of nifedipine (0.25-2.0 micrograms/kg/min), nicorandil (4.0-32.0 micrograms/kg/min) or nitroglycerin (1.0-8.0 micrograms/kg/min). Nifedipine produced a dose-related attenuation of the increase in mean arterial pressure after bolus administration of phenylephrine (from 26 +/- 1 to 7 +/- 1 mm Hg) and B-HT 933 (from 29 +/- 2 to 5 +/- 1 mm Hg). Nicorandil did not affect phenylephrine-mediated pressor responses but significantly attenuated those to B-HT 933 (28 +/- 2 to 10 +/- 1 mm Hg). In contrast, nitroglycerin had no effect on either phenylephrine or B-HT 933-mediated responses. In additional experiments after autonomic nervous system blockade, multiple doses of phenylephrine (0.6, 1.25 and 2.5 micrograms/kg i.v.) and B-HT 933 (10, 20 and 40 micrograms/kg i.v.) were administered before and after i.v. infusions of nifedipine (1.0 microgram/kg/min) or nicorandil (16.0 micrograms/kg/min). Nifedipine significantly decreased the pressor responses to all doses of phenylephrine and B-HT 933. In contrast, the attenuation from control of alpha-2-mediated increases in arterial pressure by nicorandil was partially overcome at higher doses (40 micrograms/kg) of B-HT 933.(ABSTRACT TRUNCATED AT 250 WORDS)


Adenomatoid tumor of tunica albuginea mimicking seminoma on fine needle aspiration cytology: a case report.
Adenomatoid tumor is one of the most common primary neoplasms of the paratesticular area. The epididymis is the most common site of involvement; however, it can also arise from the tunica albuginea, spermatic cord, tunica vaginalis and testis. Fine needle aspiration cytology (FNAC) is being increasingly used for lesions in the paratesticular area. Review of the literature showed very few reports on the cytologic features of adenomatoid tumor. We report the cytologic features of a case of adenomatoid tumor arising from testicular tunica albuginea. A 40-year-old man presented with right-sided scrotal swelling of 1 years' duration. FNAC of the lesion showed highly cellular smears composed of clusters of monomorphic plump cells with oval eccentric nucleus, fine granular chromatin, small single nucleolus and abundant pale to vacuolated cytoplasm. The background showed "tigroid substance," classically described in seminoma. Histopathologic examination confirmed the diagnosis as adenomatoid tumor of tunica albuginea. FNAC of an adenomatoid tumor poses a diagnostic challenge when the tumor arises from sites other than the epididymis because they can be mistaken for intratesticular tumors. It is thus important to consider this tumor in the differential diagnosis of scrotal swellings.


Presence and characterization of shiga toxin-producing Escherichia coli and other potentially diarrheagenic E. coli strains in retail meats.
To determine the presence of Shiga toxin-producing Escherichia coli (STEC) and other potentially diarrheagenic E. coli strains in retail meats, 7,258 E. coli isolates collected by the U.S. National Antimicrobial Resistance Monitoring System (NARMS) retail meat program from 2002 to 2007 were screened for Shiga toxin genes. In addition, 1,275 of the E. coli isolates recovered in 2006 were examined for virulence genes specific for other diarrheagenic E. coli strains. Seventeen isolates (16 from ground beef and 1 from a pork chop) were positive for stx genes, including 5 positive for both stx(1) and stx(2), 2 positive for stx(1), and 10 positive for stx(2). The 17 STEC strains belonged to 10 serotypes: O83:H8, O8:H16, O15:H16, O15:H17, O88:H38, ONT:H51, ONT:H2, ONT:H10, ONT:H7, and ONT:H46. None of the STEC isolates contained eae, whereas seven carried enterohemorrhagic E. coli (EHEC) hlyA. All except one STEC isolate exhibited toxic effects on Vero cells. DNA sequence analysis showed that the stx(2) genes from five STEC isolates encoded mucus-activatable Stx2d. Subtyping of the 17 STEC isolates by pulsed-field gel electrophoresis (PFGE) yielded 14 distinct restriction patterns. Among the 1,275 isolates from 2006, 11 atypical enteropathogenic E. coli (EPEC) isolates were identified in addition to 3 STEC isolates. This study demonstrated that retail meats, mainly ground beef, were contaminated with diverse STEC strains. The presence of atypical EPEC strains in retail meat is also of concern due to their potential to cause human infections.


Thrombin Upregulates PAI-1 and Mesothelial-Mesenchymal Transition Through PAR-1 and Contributes to Tuberculous Pleural Fibrosis.
Thrombin is an essential procoagulant and profibrotic mediator. However, its implication in tuberculous pleural effusion (TBPE) remains unknown. The effusion thrombin and plasminogen activator inhibitor-1 (PAI-1) levels were measured among transudative pleural effusion (TPE, n = 22) and TBPE (n = 24) patients. Pleural fibrosis, identified as radiological residual pleural thickening (RPT) and shadowing, was measured at 12-month follow-up. Moreover, in vivo and in vitro effects of thrombin on PAI-1 expression and mesothelial-mesenchymal transition (MMT) were assessed. We demonstrated the effusion thrombin levels were significantly higher in TBPE than TPE, especially greater in TBPE patients with RPT > 10mm than those without, and correlated positively with PAI-1 and pleural fibrosis area. In carbon black/bleomycin-treated mice, knockdown of protease-activated receptor-1 (PAR-1) markedly downregulated α-smooth muscle actin (α-SMA) and fibronectin, and attenuated pleural fibrosis. In pleural mesothelial cells (PMCs), thrombin concentration-dependently increased PAI-1, α-SMA, and collagen I expression. Specifically, Mycobacterium tuberculosis H37Ra (MTBRa) induced thrombin production by PMCs via upregulating tissue factor and prothrombin, and PAR-1 silencing considerably abrogated MTBRa-stimulated PAI-1 expression and MMT. Consistently, prothrombin/PAR-1 expression was evident in the pleural mesothelium of TBPE patients. Conclusively, thrombin upregulates PAI-1 and MMT and may contribute to tuberculous pleural fibrosis. Thrombin/PAR-1 inhibition may confer potential therapy for pleural fibrosis.


Using a Multi-Locus Microsatellite Typing method improved phylogenetic distribution of Candida albicans isolates but failed to demonstrate association of some genotype with the commensal or clinical origin of the isolates.
The dimorphic yeast Candida albicans is a component of the normal microflora at the mucosal surfaces of healthy individuals. It possesses an array of phenotypic properties considered as virulence traits that contribute to pathogenicity of the yeast in immuno-compromised patients. We addressed the question of the pathogenicity of lineages of C. albicans with regard to their genotype in three series of C. albicans isolates (a series of commensal isolates collected in healthy individuals, a group of bloodstream isolates and a group of non-bloodstream clinical isolates) using a Multi-Locus Microsatellite Typing (MLMT) approach based on the analysis of the polymorphism of 11 microsatellite loci. The MLMT analysis of the three series, corresponding to 174 C. albicans isolates, gave a 100% typability to the method, with a DP index of 0.999. The UPGMA analysis showed that the isolates segregated in eight phylogenetic groups. Interestingly, the clustering was comparable when using NJ and MS-tree algorithms and a good concordance index of the clustering was observed with MLST. All in all our data strongly indicated MLMT as a reliable tool for DNA-typing studies in C. albicans. Isolates from healthy and non-healthy individuals segregated at the same proportions into the eight phylogenetic groups, suggesting that isolates of different origin share the same overall pathogenicity. Surprisingly allelic frequencies at the HIS3 microsatellite differed significantly in commensal isolates (group A) from pooled groups B and C (clinical isolates), raising the possibility that some individual alleles at the HIS3 microsatellite may be associated with distinct pathogenic profiles in C. albicans.


Graphical coding data and operational guidance for implementation or modification of a LabVIEW®-based pHstat system for the cultivation of microalgae.
The influence of pH on phytoplankton physiology is an important facet of the body of research on ocean acidification. We provide data developed during the design and implementation of a novel pHstat system capable of maintaining both static and dynamic pH environments in a laboratory setting. These data both help improve functionality of the system, and provide specific coding blocks for controlling the pHstat using a LabVIEW® virtual instrument (VI). The data in this paper support the research article "Development of an economical, autonomous pHstat system for culturing phytoplankton under steady state or dynamic conditions" (Golda et al. [2]). These data will be of interest to researchers studying the effects of changing pH on phytoplankton in a laboratory context, and to those desiring to build their own pHstat system(s). These data can also be used to facilitate modification of the pHstat system to control salinity, temperature, or other environmental factors.


Inflammation and its discontents: the role of cytokines in the pathophysiology of major depression.
Recognition that inflammation may represent a common mechanism of disease has been extended to include neuropsychiatric disorders including major depression. Patients with major depression have been found to exhibit increased peripheral blood inflammatory biomarkers, including inflammatory cytokines, which have been shown to access the brain and interact with virtually every pathophysiologic domain known to be involved in depression, including neurotransmitter metabolism, neuroendocrine function, and neural plasticity. Indeed, activation of inflammatory pathways within the brain is believed to contribute to a confluence of decreased neurotrophic support and altered glutamate release/reuptake, as well as oxidative stress, leading to excitotoxicity and loss of glial elements, consistent with neuropathologic findings that characterize depressive disorders. Further instantiating the link between inflammation and depression are data demonstrating that psychosocial stress, a well-known precipitant of mood disorders, is capable of stimulating inflammatory signaling molecules, including nuclear factor kappa B, in part, through activation of sympathetic nervous system outflow pathways. Interestingly, depressed patients with increased inflammatory biomarkers have been found to be more likely to exhibit treatment resistance, and in several studies, antidepressant therapy has been associated with decreased inflammatory responses. Finally, preliminary data from patients with inflammatory disorders, as well as medically healthy depressed patients, suggest that inhibiting proinflammatory cytokines or their signaling pathways may improve depressed mood and increase treatment response to conventional antidepressant medication. Translational implications of these findings include the unique opportunity to identify relevant patient populations, apply immune-targeted therapies, and monitor therapeutic efficacy at the level of the immune system in addition to behavior.


Introducing the WHOQOL-SRPB BREF: developing a short-form instrument for assessing spiritual, religious and personal beliefs within quality of life.
The aim was to develop and conduct preliminary testing of a short-form measure to assess spiritual, religious and personal beliefs (SRPB) within quality of life (QoL). Existing data from the 132 items of the WHOQOL-SRPB (n = 5087) obtained in 18 cultures were first analysed to select the 'best' performing item from each of the eight SRPB facets. These were integrated with the 26 WHOQOL-BREF items to give 34 items in the WHOQOL-SRPB BREF. A focus group of hospital chaplains reviewed this new short-form. The WHOQOL-SRPB BREF was administered to a UK community sample (n = 230) either with an adapted WHOQOL-SRPB Importance measure or the SWBQ. A subset received both WHOQOL measures twice. Completed in 8 mins, the WHOQOL-SRPB BREF was acceptable and feasible; Importance 5.5 mins. Good internal consistency reliability was found overall (α = 0.85), for the SRPB domain (α = 0.83), and Importance (α = 0.90). Domains were moderately correlated. Domain test-retest reliability was acceptable in both WHOQOL measures, except for SRPB Importance. Sleep was linked with religious beliefs. Hope and wholeness were widely associated with non-spiritual facets. Factor analysis (maximum likelihood) of items largely confirmed the WHOQOL domain structure, adding SRPB as a significant fifth domain. Internally, SRPB distinguished religious from existential beliefs, and was validated by association with personal and transcendental well-being from the SWBQ. Preliminary evidence shows that the WHOQOL-SRPB BREF is sound for use in, and beyond health care. Extracted from a measure already available in 18 languages, this short-form can be immediately used where such translations exist.


Structure of the fungal beta-glucan-binding immune receptor dectin-1: implications for function.
The murine molecule dectin-1 (known as the beta-glucan receptor in humans) is an immune cell surface receptor implicated in the immunological defense against fungal pathogens. Sequence analysis has indicated that the dectin-1 extracellular domain is a C-type lectin-like domain, and functional studies have established that it binds fungal beta-glucans. We report several dectin-1 crystal structures, including a high-resolution structure and a 2.8 angstroms resolution structure in which a short soaked natural beta-glucan is trapped in the crystal lattice. In vitro characterization of dectin-1 in the presence of its natural ligand indicates higher-order complex formation between dectin-1 and beta-glucans. These combined structural and biophysical data considerably extend the current knowledge of dectin-1 structure and function, and suggest potential mechanisms of defense against fungal pathogens.


A flexible framework for sparse simultaneous component based data integration.
High throughput data are complex and methods that reveal structure underlying the data are most useful. Principal component analysis, frequently implemented as a singular value decomposition, is a popular technique in this respect. Nowadays often the challenge is to reveal structure in several sources of information (e.g., transcriptomics, proteomics) that are available for the same biological entities under study. Simultaneous component methods are most promising in this respect. However, the interpretation of the principal and simultaneous components is often daunting because contributions of each of the biomolecules (transcripts, proteins) have to be taken into account. We propose a sparse simultaneous component method that makes many of the parameters redundant by shrinking them to zero. It includes principal component analysis, sparse principal component analysis, and ordinary simultaneous component analysis as special cases. Several penalties can be tuned that account in different ways for the block structure present in the integrated data. This yields known sparse approaches as the lasso, the ridge penalty, the elastic net, the group lasso, sparse group lasso, and elitist lasso. In addition, the algorithmic results can be easily transposed to the context of regression. Metabolomics data obtained with two measurement platforms for the same set of Escherichia coli samples are used to illustrate the proposed methodology and the properties of different penalties with respect to sparseness across and within data blocks. Sparse simultaneous component analysis is a useful method for data integration: First, simultaneous analyses of multiple blocks offer advantages over sequential and separate analyses and second, interpretation of the results is highly facilitated by their sparseness. The approach offered is flexible and allows to take the block structure in different ways into account. As such, structures can be found that are exclusively tied to one data platform (group lasso approach) as well as structures that involve all data platforms (Elitist lasso approach). The additional file contains a MATLAB implementation of the sparse simultaneous component method.


Biliary stents are forgotten more frequently in elderly patients
Plastic biliary stents that remain in situ for more than 12 months, called forgotten biliary stents (FBSs), can cause complications such as cholangitis, stent migration, stent occlusion, and perforation. The medical records of patients who underwent ERCP procedures from December 2016 to December 2020 were analysed retrospectively. Data on patient characteristics, indications for ERCP and stenting, stent types, stenting duration, complications, and causes of FBSs were obtained from the hospital's database. A total of 48 cases with FBSs were analysed. The mean age (SD) of the patients was 71.23 years (±12.165), the male-to-female ratio was 23/25 (0.92), and the mean stenting duration was 27.12 months (range: 12–84 months). The most common indication for biliary stenting was irretrievable choledochal stones (40/48). Stone formation (79%) and proximal stent migration (26.4%) were the most frequent complications. The patients in the FBS group were significantly older than those from whom stents were removed in a timely manner (71.23 vs. 62.43 years, p < 0.001). Endoscopic treatment was possible in all cases; surgery was not required in any case. The most common cause of FBSs cited by patients was not having been informed about the need for long-term management of their stents (n = 14, 29.2%) FBSs are potentially problematic particularly in elderly patients. Communication with the patient to remind them of the need for stent management is important for preventing FBSs.


Establishment of four new mesothelioma cell lines: characterization by ultrastructural and immunophenotypic analysis.
The aim of this study was to assess the biological characteristics of four new malignant mesothelioma (MM) cell lines. Since simian virus (SV)40 sequences have been recently detected in MM, SV40 large T antigen (Tag) expression was also analysed. MM cell lines were characterized by morphological, ultrastructural and cytogenetic analysis. Expression of Tag and of relevant MM markers was studied by immunocytochemistry, surface antigens by indirect immunofluorescence and immunomodulating cytokines by enzyme-linked immunosorbent assay (ELISA). The four MM cell lines, established from pleural effusions, showed a slow proliferation rate and pleomorphic changes during culture. Cell lines expressed vimentin, cytokeratins 8 and 18, and the mesothelial antigen recognized by HBME-1 monoclonal antibody, but not carcinoembryonic antigen. Surface human leukocyte antigen (HLA)-class I and intercellular adhesion molecule (ICAM)-1 molecules were present on all the cell lines. While HLA class II and CD86 were constitutively undetectable, HLA-class II was present after interferon (IFN)-gamma stimulation. All cell lines displayed abnormal karyotypes with chromosome 6 abnormalities. Transforming growth factor (TGF)-beta2 and interleukin (IL)-6 were constitutively secreted, while tumour necrosis factor (TNF)-alpha was secreted only in response to lipopolysaccharide. Intranuclear Tag was expressed in two cell lines. The persistence of large T antigen with human leukocyte antigen class I and intercellular adhesion molecule-1 positivity may point to large T antigen as a target for cytotoxic T-lymphocyte-based immunotherapy in some malignant mesothelioma patients.


Achievements and future problems with component surgery of the cornea.
Penetrating keratoplasty (PKP) has long been the standard procedure for treating corneal opacification. Recent advances in corneal surgery have enabled us to perform component surgery of the cornea, replacing necessary cells and tissue only instead of changing the entire layer of the cornea. Corneal components can be transplanted as lamellar sections of donor cornea or as ex vivo expanded cell sheets with or without biologic carriers. Transplantation of cultivated epithelial sheets expanded from limbal epithelium or oral mucosal epithelium, deep lamellar keratoplasty (DLKP), and deep lamellar endothelial keratoplasty (DLEK) are already in clinical application. These surgical techniques have the advantage of reducing surgical trauma, risk of immunologic rejection, and refractive error. Furthermore, severe ocular surface diseases caused by total limbal stem-cell deficiency including Stevens-Johnson syndrome can be treated by such epithelial sheet transplantation. Although limbal transplantation (LT) has also been applied for such cases, epithelial sheet transplantation has the advantage of covering the corneal surface during surgery, which may reduce postoperative inflammation. However, the success rate of clinical outcome is still not sufficient. We review the surgical technique of component surgery and compare the clinical results including visual acuity, clarity of the cornea, and neovascularization between LT and epithelial sheet transplantation and among PKP, DLKP, and DLEK in our institute. Further refinements in surgical and biologic technology may take the limits of corneal regenerative medicine to new horizons.


Label-free phenotyping of peripheral blood lymphocytes by infrared imaging.
It is now widely accepted that the immune microenvironment of tumors and more precisely Tumor Infiltrating Lymphocytes (TIL) play an important role in cancer development and outcome. TILs are considered to be important prognostic and predictive factors based on a growing body of clinical evidence; however, their presence at the tumor site is not currently assessed routinely. FTIR (Fourier transform infrared) imaging has proven it has value in studying a range of tumors, particularly for characterizing tumor cells. Currently, very little is known about the potential for FTIR imaging to characterize TIL. The present proof of concept study investigates the ability of FTIR imaging to identify the principal lymphocyte subpopulations present in human peripheral blood (PB). A negative cell isolation method was employed to select pure, label-free, helper T cells (CD4(+)), cytotoxic T cells (CD8(+)) and B cells (CD19(+)) from six healthy donors PB by Fluorescence Activated Cell Sorting (FACS). Cells were centrifuged onto Barium Fluoride windows and ten infrared images were recorded for each lymphocyte subpopulation from all six donors. After spectral pre-treatment, statistical analyses were performed. Unsupervised Principal Component Analyses (PCA) revealed that in the absence of donor variability, CD4(+) T cells, CD8(+) T cells and B cells each display distinct IR spectral features. Supervised Partial Least Square Discriminant Analyses (PLS-DA) demonstrated that the differences between the three lymphocyte subpopulations are reflected in their IR spectra, permitting their individual identification even when significant donor variability is present. Our results also show that a distinct spectral signature is associated with antibody binding. To our knowledge this is the first study reporting that FTIR imaging can effectively identify T and B lymphocytes and differentiate helper T cells from cytotoxic T cells. This proof of concept study demonstrates that FTIR imaging is a reliable tool for the identification of lymphocyte subpopulations and has the potential for use in characterizing TIL.


Molecular characterisation of the early response in pigs to experimental infection with Actinobacillus pleuropneumoniae using cDNA microarrays.
The bacterium Actinobacillus pleuropneumoniae is responsible for porcine pleuropneumonia, a widespread, highly contagious and often fatal respiratory disease of pigs. The general porcine innate immune response after A. pleuropneumoniae infection is still not clarified. The objective of this study was hence to characterise the transcriptional response, measured by using cDNA microarrays, in pigs 24 hours after experimental inoculation with A. pleuropneumoniae. Microarray analyses were conducted to reveal genes being differentially expressed in inflamed versus non-inflamed lung tissue sampled from inoculated animals as well as in liver and tracheobronchial lymph node tissue sampled from three inoculated animals versus two non-inoculated animals. The lung samples were studied using a porcine cDNA microarray with 5375 unique PCR products while liver tissue and tracheobronchial lymph node tissue were hybridised to an expanded version of the porcine microarray with 26879 unique PCR products. A total of 357 genes differed significantly in expression between infected and non-infected lung tissue, 713 genes differed in expression in liver tissue from infected versus non-infected animals and 130 genes differed in expression in tracheobronchial lymph node tissue from infected versus non-infected animals. Among these genes, several have previously been described to be part of a general host response to infections encoding immune response related proteins. In inflamed lung tissue, genes encoding immune activating proteins and other pro-inflammatory mediators of the innate immune response were found to be up-regulated. Genes encoding different acute phase reactants were found to be differentially expressed in the liver. The obtained results are largely in accordance with previous studies of the mammalian immune response. Furthermore, a number of differentially expressed genes have not previously been associated with infection or are presently unidentified. Determination of their specific roles during infection may lead to a better understanding of innate immunity in pigs. Although additional work including more animals is clearly needed to elucidate host response to porcine pleuropneumonia, the results presented in this study demonstrate three subsets of genes consistently expressed at different levels depending upon infection status.


Chronic obstructive pulmonary disease and subsequent overall and lung cancer mortality in low-income adults.
Chronic obstructive pulmonary disease (COPD) is a known risk factor for lung cancer and a leading cause of mortality in the U.S., but its impact may not be fully appreciated, especially among low-income populations in the southeast where COPD prevalence and lung cancer incidence are elevated. We conducted a prospective study among 26,927 low-income adults age 40-79 in the Southern Community Cohort Study who had a Center for Medicare and Medicaid Services (CMS) encounter prior to enrollment and were followed for a median of over 6 years. Using a validated algorithm for assessing COPD from CMS claims data, we estimated COPD prevalence and potential misreporting. From Cox proportional hazard models, we computed overall and lung cancer-specific mortality according to COPD status. The overall prevalence of CMS-diagnosed COPD was 16%, but was twice as high among whites as blacks. Only 35% of these individuals, however, self-reported having COPD, with underreporting significantly greater for blacks than whites. Smoking-adjusted all-cause mortality was increased by 1.7-fold and lung cancer mortality by 2.3-fold among those with a CMS COPD diagnosis, with similar patterns in blacks and whites, but no excess was found among those self-reporting COPD and without CMS confirmation. The prevalence of COPD in this low-income population may be greater than previously recognized and misreporting is common. COPD is associated with elevated lung cancer mortality, even among those not self-reporting the condition.


Asymmetric Orientation Distribution Functions (AODFs) revealing intravoxel geometry in diffusion MRI.
Characterization of anisotropy via diffusion MRI reveals fiber crossings in a substantial portion of voxels within the white-matter (WM) regions of the human brain. A considerable number of such voxels could exhibit asymmetric features such as bends and junctions. However, widely employed reconstruction methods yield symmetric Orientation Distribution Functions (ODFs) even when the underlying geometry is asymmetric. In this paper, we employ inter-voxel directional filtering approaches through a cone model to reveal more information regarding the cytoarchitectural organization within the voxel. The cone model facilitates a sharpening of the ODFs in some directions while suppressing peaks in other directions, thus yielding an Asymmetric ODF (AODF) field. We also show that a scalar measure of AODF asymmetry can be employed to obtain new contrast within the human brain. The feasibility of the technique is demonstrated on in vivo data obtained from the MGH-USC Human Connectome Project (HCP) and Parkinson's Progression Markers Initiative (PPMI) Project database. Characterizing asymmetry in neural tissue cytoarchitecture could be important for localizing and quantitatively assessing specific neuronal pathways.


Rapid Decrease in KRT14 and TP53 mRNA Expression in the Buccal Mucosa of Patients Receiving Total-Body Irradiation for Allogeneic Stem Cell Transplantation.
The only curative treatment option for relapsed patients with acute myeloid leukemia (AML) is allogeneic stem cell transplantation. Depletion of hematopoietic stem cells and leukemic blast cells is achieved through the systemic administration of DNA damaging agents, including total-body irradiation (TBI) prior to transplantation. Since other tissues are radiosensitive, the identification of biomarkers could facilitate the management of additional toxicities. Buccal keratinocytes are readily accessible and could provide a source of cells for RNA analysis. In this study, we obtained miRNAs and mRNAs from daily buccal swabs collected from patients undergoing allogeneic stem cell transplantation. Unexpectedly, there was no prominent p53-induced mRNA or miRNA response in these samples, despite the fact that the p53 pathway is a well-characterized radiation-inducible response. Instead, the expression of mRNAs encoding p53 and cytokeratin 14 (TP53 and KRT14, respectively) decreased precipitously within hours of the first radiation treatment. These patients went on to develop oral mucositis, however, it is unclear whether TP53 and/or KRT14 expression are predictive of this adverse event. Larger scale analysis of buccal epithelial samples from patients undergoing allogeneic stem cell transplantation appears to be warranted.


Sanguis draconis, a dragon's blood resin, attenuates high glucose-induced oxidative stress and endothelial dysfunction in human umbilical vein endothelial cells.
Hyperglycaemia, a characteristic feature of diabetes mellitus, induces endothelial dysfunction and vascular complications by limiting the proliferative potential of these cells. Here we aimed to investigate the effect of an ethanolic extract of Sanguis draconis (SD), a kind of dragon's blood resin that is obtained from Daemonorops draco (Palmae), on human umbilical vein endothelial cells (HUVEC) under high-glucose (HG) stimulation and its underlying mechanism. Concentration-dependent (0-50 μg/mL) assessment of cell viability showed that SD does not affect cell viability with a similar trend up to 48 h. Remarkably, SD (10-50 μg/mL) significantly attenuated the high-glucose (25 and 50 mM) induced cell toxicity in a concentration-dependent manner. SD inhibited high glucose-induced nitrite (NO) and lipid peroxidation (MDA) production and reactive oxygen species (ROS) formation in HUVEC. Western blot analysis revealed that SD treatments abolished HG-induced phosphorylation of extracellular signal-regulated kinase 1/2 (ERK 1/2), nuclear transcription factor, κB (NF-κB), VCAM-1, and E-selectin, and it also blocked the breakdown of PARP-116 kDa protein in a dose-dependent manner. Furthermore, we found that SD increased the expression of Bcl-2 and decreased Bax protein expression in HG-stimulated HUVEC. Thus, these results of this study demonstrate for the first time that SD inhibits glucose induced oxidative stress and vascular inflammation in HUVEC by inhibiting the ERK/NF-κB/PARP-1/Bax signaling cascade followed by suppressing the activation of VCAM-1 and E-selectin. These data suggest that SD may have a therapeutic potential in vascular inflammation due to the decreased levels of oxidative stress, apoptosis, and PARP-1 activation.


Persistent fluctuations in stride intervals under fractal auditory stimulation.
Stride sequences of healthy gait are characterized by persistent long-range correlations, which become anti-persistent in the presence of an isochronous metronome. The latter phenomenon is of particular interest because auditory cueing is generally considered to reduce stride variability and may hence be beneficial for stabilizing gait. Complex systems tend to match their correlation structure when synchronizing. In gait training, can one capitalize on this tendency by using a fractal metronome rather than an isochronous one? We examined whether auditory cues with fractal variations in inter-beat intervals yield similar fractal inter-stride interval variability as isochronous auditory cueing in two complementary experiments. In Experiment 1, participants walked on a treadmill while being paced by either an isochronous or a fractal metronome with different variation strengths between beats in order to test whether participants managed to synchronize with a fractal metronome and to determine the necessary amount of variability for participants to switch from anti-persistent to persistent inter-stride intervals. Participants did synchronize with the metronome despite its fractal randomness. The corresponding coefficient of variation of inter-beat intervals was fixed in Experiment 2, in which participants walked on a treadmill while being paced by non-isochronous metronomes with different scaling exponents. As expected, inter-stride intervals showed persistent correlations similar to self-paced walking only when cueing contained persistent correlations. Our results open up a new window to optimize rhythmic auditory cueing for gait stabilization by integrating fractal fluctuations in the inter-beat intervals.


Dual-Process Theories of Higher Cognition: Advancing the Debate.
Dual-process and dual-system theories in both cognitive and social psychology have been subjected to a number of recently published criticisms. However, they have been attacked as a category, incorrectly assuming there is a generic version that applies to all. We identify and respond to 5 main lines of argument made by such critics. We agree that some of these arguments have force against some of the theories in the literature but believe them to be overstated. We argue that the dual-processing distinction is supported by much recent evidence in cognitive science. Our preferred theoretical approach is one in which rapid autonomous processes (Type 1) are assumed to yield default responses unless intervened on by distinctive higher order reasoning processes (Type 2). What defines the difference is that Type 2 processing supports hypothetical thinking and load heavily on working memory.


Quantitative characterization of capsaicin-induced TRPV1 ion channel activation in HEK293 cells by impedance spectroscopy.
The analysis of receptor activity, especially in its native cellular environment, has always been of great interest to evaluate its intrinsic but also downstream biological activity. An important group of cellular receptors are ion channels. Since they are involved in a broad range of crucial cell functions, they represent important therapeutic targets. Thus, novel analytical techniques for the quantitative monitoring and screening of biological receptor activity are of great interest. In this context, we developed an impedance spectroscopy-based label-free and non-invasive monitoring system that enabled us to analyze the activation of the transient receptor potential channel Vanilloid 1 (TRPV1) in detail. TRPV1 channel activation by capsaicin resulted in a reproducible impedance decrease. Moreover, concentration response curves with an EC50 value of 0.9 μM could be determined. Control experiments with non TRPV1 channel expressing HEK cells as well as experiments with the TRPV1 channel blocker ruthenium red validated the specificity of the observed impedance decrease. More strikingly, through correlative studies with a cytoskeleton restructuring inhibitor mixture and equivalent circuit analysis of the acquired impedance spectra, we could quantitatively discriminate between the direct TRPV1 channel activation and downstream-induced biological effects. In summary, we developed a quantitative impedimetric monitoring system for the analysis of TRPV1 channel activity as well as downstream-induced biological activity in living cells. It has the capabilities to identify novel ion channel activators as well as inhibitors for the TRPV1 channel but could also easily be applied to other ion channel-based receptors.


Analysis of putative apoplastic effectors from the nematode, Globodera rostochiensis, and identification of an expansin-like protein that can induce and suppress host defenses.
The potato cyst nematode, Globodera rostochiensis, is an important pest of potato. Like other pathogens, plant parasitic nematodes are presumed to employ effector proteins, secreted into the apoplast as well as the host cytoplasm, to alter plant cellular functions and successfully infect their hosts. We have generated a library of ORFs encoding putative G. rostochiensis putative apoplastic effectors in vectors for expression in planta. These clones were assessed for morphological and developmental effects on plants as well as their ability to induce or suppress plant defenses. Several CLAVATA3/ESR-like proteins induced developmental phenotypes, whereas predicted cell wall-modifying proteins induced necrosis and chlorosis, consistent with roles in cell fate alteration and tissue invasion, respectively. When directed to the apoplast with a signal peptide, two effectors, an ubiquitin extension protein (GrUBCEP12) and an expansin-like protein (GrEXPB2), suppressed defense responses including NB-LRR signaling induced in the cytoplasm. GrEXPB2 also elicited defense response in species- and sequence-specific manner. Our results are consistent with the scenario whereby potato cyst nematodes secrete effectors that modulate host cell fate and metabolism as well as modifying host cell walls. Furthermore, we show a novel role for an apoplastic expansin-like protein in suppressing intra-cellular defense responses.


Combination of microsurgery and gene therapy for spinal dorsal root injury repair.
Brachial plexus injury is frequent after traffic accident in adults or shoulder dystocia in newborns. Whereas surgery can restore arm movements, therapeutic options are missing for sensory defects. Dorsal root (DR) ganglion neurons convey sensory information to the central nervous system (CNS) through a peripheral and a central axon. Central axons severed through DR section or avulsion during brachial plexus injury inefficiently regenerate and do not reenter the spinal cord. We show that a combination of microsurgery and gene therapy circumvented the functional barrier to axonal regrowth at the peripheral and CNS interface. After cervical DR section in rats, microsurgery restored anatomical continuity through a nerve graft that laterally connected the injured DR to an intact DR. Gene transfer to cells in the nerve graft induced the local release of neurotrophin-3 (NT-3) and glial cell line-derived neurotrophic factor (GDNF) and stimulated axonal regrowth. Central DR ganglion axons efficiently regenerated and invaded appropriate areas of the spinal cord dorsal horn, leading to partial recovery of nociception and proprioception. Microsurgery created conditions for functional restoration of DR ganglion central axons, which were improved in combination with gene therapy. This combination treatment provides means to reduce disability due to somatosensory defects after brachial plexus injury.


Treatment of Liver Metastases Using an Internal Target Volume Method for Stereotactic Body Radiotherapy.
The prognosis of patients with metastatic cancers has improved in the past decades due to effective chemotherapy and oligometastatic surgery. For inoperable patients, local ablation therapies, such as stereotactic body radiotherapy (SBRT), can provide effective local tumor control with minimal toxicity. Because of its high precision and accuracy, SBRT delivers a higher radiation dose per fraction, is more effective, and targets smaller irradiation volumes than does conventional radiotherapy. In addition, steep dose gradients from target lesions to surrounding normal tissues are achieved using SBRT; thus, SBRT provides more effective tumor control and exhibits fewer side effects than conventional radiotherapy. The use of SBRT is prevalent for treating intracranial lesions (known as stereotactic radiosurgery); however, it is now also used for treating spinal and adrenal metastases. Because of advancements in image-guided assistance and respiratory motion management, several studies have investigated the use of SBRT for treating lung or liver tumors, which move as a patient breathes. The results of these studies have suggested that SBRT favorably controls tumors in the case of moving lesions. Four-dimensional computed tomography (4D-CT) with an abdominal compressor (AC) is clinically convenient for effective respiratory motion management. Because this method is noninvasive and allows free breathing, its use reduces complications. Furthermore, patients consider this method convenient. Moreover, it is considered more efficient than other methods of respiratory motion management by physicians and therapists. The use of 4D-CT with an AC for treating pulmonary lesions has also been widely investigated, and the technique is gaining acceptance for treating hepatic lesions. However, the protocols for using 4D-CT with an AC for treating hepatic lesions are different from those used for treating pulmonary lesions. In this article, we describe a new protocol for SBRT with 4D-CT and an AC for treating liver metastases.


Organometric investigations of the spleen and liver by ultrasound in Schistosoma mansoni endemic and nonendemic villages in Senegal.
With the intention of ultrasonographically assessing hepatosplenic morbidity in Schistosoma mansoni infection and of validating the grading system applied (Cairo classification), 191 subjects in a schistosomiasis endemic village and 247 controls from a nonendemic village in northern Senegal underwent sonographic examination of the liver and spleen. Measurements of the diameters of the peripheral periportal vein branches, the main portal vein stem, liver size (left lobe and right lobe), and spleen length in the endemic village were compared with those in the nonendemic village to evaluate the much discussed influence of S. mansoni infection on those variables. To subtract this presumed influence from reference values for the named variables, they are given as measured in the nonendemic village, stratified by body weight, enabling future investigators on schistosomiasis-induced morbidity to refer to these reference values. The 95th percentile regarding peripheral periportal vein branch diameter in the control groups was exceeded in 24% of the subjects in the endemic group. It was exceeded by 6% for the main portal vein stem diameter, 13% for the left liver lobe, 12% for the right liver lobe, and 14% for the spleen length. According to the Cairo classification, 97% of the endemic population and 81% of the controls had periportal thickening of the liver, mostly grade I. We conclude that 1) hepatic morbidity in the S. mansoni endemic area was low, despite strikingly high intensities of infection; 2) the Cairo classification in its present form overestimates periportal thickening, especially in the case of mild morbidity; and 3) body height-dependent reference values, obtained from endemic controls, must be applied for organometric parameters.


Tensions and opportunities in social prescribing. Developing a framework to facilitate its implementation and evaluation in primary care: a realist review.
Social prescribing (SP) involves linking patients in primary care with services provided by the voluntary and community sector (VCS). Despite growing interest within NHS primary care, it remains unclear how and under what circumstances SP might contribute to good practice. To define 'good' practice in SP by identifying context-specific enablers and tensions. To contribute to the development of an evidence-based framework for theorising and evaluating SP within primary care. Realist review of secondary data from primary care-based SP schemes. Academic articles and grey literature were searched for qualitative and quantitative evidence following the Realist And Meta-narrative Evidence Syntheses - Evolving Standards (RAMESES). Common SP practices were characterised in three settings (general practice, link workers, and community sector) using archetypes that ranged from best to worst practice. A total of 140 studies were included for analysis. Resources were identified influencing the type and potential impact of SP practices and four dimensions were outlined in which opportunities for good practice arise: 1) individual characteristics (stakeholder's buy-in, vocation, and knowledge); 2) interpersonal relations (trustful, bidirectional, informed, supportive, and transparent and convenient interactions within and across sectors); 3) organisational contingencies (the availability of a predisposed practice culture, leadership, training opportunities, supervision, information governance, resource adequacy, accessibility, and continuity of care within organisations); and 4) policy structures (bottom-up and coherent policymaking, stable funding, and suitable monitoring strategies). Findings were synthesised in a multilevel, dynamic, and usable SP framework. The realist review and resulting framework revealed that SP is not inherently advantageous. Specific individual, interpersonal, organisational, and policy resources are needed to ensure SP best practice in primary care.


Oxygen supply limits the heat tolerance of avian embryos.
Physiologists have primarily focused on two potential explanations for heat stress in animals-the classic model of molecular stability and an alternative model of oxygen limitation. Although the classic model has widespread support, the oxygen-supply model applies to many aquatic animals and some terrestrial ones. In particular, the embryonic stage of terrestrial animals seems most susceptible to oxygen limitation because embryos acquire oxygen from the atmosphere by diffusion rather than ventilation. We report experiments confirming the two conditions of the oxygen-supply model in Japanese quail embryos, Coturnix coturnix. Hypoxia (12% O2) greatly reduced the chance of survival at 47.5°C, and hyperoxia greatly improved the chance of survival at 48.5°C. This finding expands the scope of the oxygen-supply model to a terrestrial, endothermic species, suggesting that oxygen supply generally limits the heat tolerance of embryos.


Mental distress and service utilization among help-seeking, community-dwelling older adults.
This study aimed to characterize healthcare and human services utilization among mentally distressed and non-distressed clients receiving in-home care management assessment by aging services provider network (ASPN) agencies in the U.S.A. A two-hour research interview was administered to 378 English-speaking ASPN clients aged 60+ years in Monroe County, NY. A modified Cornell Services Index measured service utilization for the 90 days prior to the ASPN assessment. Clients with clinically significant anxiety or depressive symptoms were considered distressed. ASPN clients utilized a mean of 2.93 healthcare and 1.54 human services. The 42% of subjects who were distressed accessed more healthcare services (e.g. mental health, intensive medical services) and had more outpatient visits and days hospitalized than the non-distressed group. Contrary to expectations, distressed clients did not receive more human services. Among those who were distressed, over half had discussed their mental health with a medical professional in the past year, and half were currently taking a medication for their emotional state. A far smaller proportion had seen a mental health professional. In the U.S.A., aging services providers serve a population with high medical illness burden and medical service utilization. Many clients also suffer from anxiety and depression, which they often have discussed with a medical professional and for which they are receiving medications. Few, however, have seen a mental health specialist preceding intake by the ASPN agency. Optimal care for this vulnerable, service intensive group would integrate primary medical and mental healthcare with delivery of community-based social services for older adults.


Parallel modified signed-digit arithmetic using an optoelectronic shared content-addressable-memory processor.
Addition is the most primitive arithmetic operation in digital computation. Other arithmetic operations such as subtraction, multiplication, and division can all be performed by addition together with some logic operations. With the binary number system, addition speed is inevitably limited by the carry-propagation schemes. On the other hand, carry-free addition is possible when the modified signed-digit (MSD) number representation is used. We propose a novel optoelectronic scheme to handle the parallel MSD addition and subtraction operations. An optoelectronic shared content-addressable memroy is introduced. The shared content-addressable memory uses free-space optical processing to handle the large amount of parallel memory access operations and uses electronics to postprocess and derive logic decisions. We analyze the accuracy that the required optical hardware can deliver by using a statistical cross-talk-rate model that we propose. We also evaluate other important device and system performanceparameters, such as the memory capacity or the maximum number of parallel bits the adder can handle in terms of a given cross-talk rate at a certain repetition rate, the corresponding diffraction-limited memory density, and the system's power efficiency. To confirm the underlining operational principles of the proposed optoelectronic shared content-addressable-memory MSD adder, we design and perform initial experiments for handling 8-bit MSD number addition and subtraction and present the results.


The availability of fast-food and full-service restaurants in the United States: associations with neighborhood characteristics.
Parallel to the rising obesity epidemic, food consumption patterns and household expenditures show a marked upward trend in total energy intake derived from away-from-home sources. This study conducted cross-sectional multivariate analyses to examine associations between local-area racial, ethnic, and income characteristics and the availability of full-service and fast-food restaurants. Based on a U.S. national census of 28,050 ZIP codes that cover a population of 280,675,874 people, restaurant outlet data were linked to 2000 Census Bureau data based on ZIP code tabulation areas and analyses were undertaken using negative binomial count models and ordinary least squares regression analyses. Study results showed that higher- versus lower-income, predominantly black and racially mixed versus predominantly white and Hispanic versus non-Hispanic neighborhoods had fewer available full-service and fast-food restaurants. Near-low- and middle-income neighborhoods had the highest number of available restaurants with 1.24 and 1.22 times number of full-service restaurants and 1.34 and 1.28 times the number of fast-food restaurants compared to high-income neighborhoods. Predominantly black neighborhoods were found to have 58.2% and 59.3% of the number of full-service and fast-food restaurants available in predominantly white neighborhoods. No statistically significant differences were found in the relative availability of fast-food versus full-service restaurants by income, race, or ethnicity in the national sample used. However, across urban areas, near-low-, middle-, and near-high- versus high-income neighborhoods and predominantly black versus white neighborhoods were found to have moderately higher proportions of fast-food among total restaurants. In urban areas, higher proportions of available fast-food restaurants out of total restaurants in predominantly black versus predominantly white neighborhoods may contribute to racial differences in obesity rates.


The Prognostic Value of Cardiovascular Risk Factors and Laboratory Biomarkers in Predicting 6-Month Outcomes in High-risk Patients with Non-ST Segment Elevation Myocardial Infarction.
Acute coronary syndrome (ACS) represents a spectrum of ischemic myocardial disease including unstable angina (UA), non-ST-segment elevation myocardial infarction (NSTEMI), and ST-elevation myocardial infarction (STEMI). Various prognostic scores were developed for patients presenting with NSTEMI-ACS. Among these scores, the GRACE risk score offers the best discriminative performance for prediction of in-hospital and 6-month mortality. However, the GRACE score is limited and cannot be used in several ethnic populations. Moreover, it is not predictive of clinical outcomes other than mortality. To assess the prognostic value of traditional cardiovascular risk factors and laboratory biomarkers in predicting 6-month major adverse cardiac and cerebrovascular events (MACCE), including hospitalization, recurrent percutaneous coronary intervention (PCI), stroke, and cardiovascular mortality in patients with NSTEMI treated with PCI. This retrospective study included consecutive patients admitted with an initial diagnosis of NSTEMI to the cardiac intensive care unit (CICU) at the Tzafon Medical Center, Israel, between April 2015 and August 2018 and treated by PCI within 48 hours of admission. A total of 223 consecutive patients with NSTEMI treated by PCI were included in the study. Logarithmebrain natriuretic peptide (LogₑBNP), prior MI, and Hb levels were found to be significant predictors of any first MACCE. Only logₑBNP was found to be an independent predictor of a first MACCE event by multivariate logistic regression analysis. LogₑBNP is an independent predictor of worse prognosis in patients with NSTEMI. Routine evaluation of BNP levels should be considered in patients admitted with NSTEMI.


Broad-host-range expression vectors with tightly regulated promoters and their use to examine the influence of TraR and TraM expression on Ti plasmid quorum sensing.
Experiments requiring strong repression and precise control of cloned genes can be difficult to conduct because of the relatively high basal level of expression of currently employed promoters. We report the construction of a family of vectors that contain a reengineered lacI(q)-lac promoter-operator complex in which cloned genes are strongly repressed in the absence of inducer. The vectors, all based on the broad-host-range plasmid pBBR1, are mobilizable and stably replicate at moderate copy number in representatives of the alpha- and gammaproteobacteria. Each vector contains a versatile multiple cloning site that includes an NdeI site allowing fusion of the cloned gene to the initiation codon of lacZalpha. In each tested bacterium, a uidA reporter fused to the promoter was not expressed at a detectable level in the absence of induction but was inducible by 10- to 100-fold, depending on the bacterium. The degree of induction was controllable by varying the concentration of inducer. When the vector was tested in Agrobacterium tumefaciens, a cloned copy of the traR gene, the product of which is needed at only a few copies per cell, did not confer activity under noninducing conditions. We used this attribute of very tight and variably regulatable control to assess the relative amounts of TraR required to activate the Ti plasmid conjugative transfer system. We identified levels of induction that gave wild-type transfer frequencies, as well as levels that induced correspondingly lower frequencies of transfer. We also used this system to show that the antiactivator TraM sets the level of intracellular TraR required for tra gene activation.


Human T-cell leukemia virus type 1 envelope glycoprotein gp46 interacts with cell surface heparan sulfate proteoglycans.
The major receptors required for attachment and entry of the human T-cell leukemia virus type 1 (HTLV-1) remain to be identified. Here we demonstrate that a functional, soluble form of the HTLV-1 surface envelope glycoprotein, gp46, fused to an immunoglobulin Fc region (gp46-Fc) binds to heparan sulfate proteoglycans (HSPGs) on mammalian cells. Substantial binding of gp46-Fc to HeLa and Chinese hamster ovary (CHO) K1 cells that express HSPGs was detected, whereas binding to the sister CHO lines 2244, which expresses no HSPGs, and 2241, which expresses no glycosaminoglycans (GAGs), was much reduced. Enzymatic removal of HSPGs from HeLa and CHO K1 cells also reduced gp46-Fc binding. Dextran sulfate inhibited gp46-Fc binding to HSPG-expressing cells in a dose-dependent manner, whereas chondroitin sulfate was less effective. By contrast, dextran sulfate inhibited gp46-Fc binding to GAG-negative cells such as CHO 2244, CHO 2241, and Jurkat T cells weakly or not at all. Dextran sulfate inhibited HTLV-1 envelope glycoprotein (Env)-pseudotyped virus infection of permissive, HSPG-expressing target cells and blocked syncytium formation between HTLV-1 Env-expressing cells and HSPG-expressing permissive target cells. Finally, HSPG-expressing cells were more permissive for HTLV-1 Env-pseudotyped virus infection than HSPG-negative cells. Thus, similar to other pathogenic viruses, HTLV-1 may have evolved to use HSPGs as cellular attachment receptors to facilitate its propagation.


Analysis of the barriers of mental distress disclosure in medical inpatients in Taiwan.
Disclosure of mental distress to physicians is important for mental illness identification, early referrals and proper treatment to prevent suicide. Little is known about what affects mental health communication in the clinical settings in the Chinese societies. The study aimed to identify the demographic, psychosocial and medical factors related to people with non-disclosure of their mental distress. We interviewed a series of consecutive inpatients from two medical wards of a general hospital in northern Taiwan. We collected depressive symptoms (the Patient Health Questionnaire, PHQ-9), living arrangement, threatening life events, suicide risks (i.e. past self-harm history, lifetime suicide ideas and hopelessness) and recent experience of mental distress disclosure. Furthermore, we explored the reasons of non-disclosure. A total of 230 medical inpatients agreed to participate (53.5% males). The results indicated that only 5.7% actually communicated their mental health concerns, and that 12.3% were willing to talk about their mental health problems. Among the 69 (30%) depressed participants (PHQ-9 score ≥ 10 points), the disclosure rate was low (8.7%) and the wish to talk about mental distress was also low (10.3%). After adjustment, living alone (OR = 7.58, 95% CI = 1.56-36.91) and having stressful life events (OR = 3.68, 95% CI = 1.09-12.46) remained significant in predicting disclosure of mental distress. The 109 participants attributed their refusal of communicating mental distress as medical-related attributes, subjective perceptions or sociocultural factors. Communication of mental distress in medical settings was uncommon due to medical or psychosocial barriers in Taiwan. Skill training to facilitate disclosure in medical education and public campaigns to improve knowledge of depression and enhance help-seeking deserve more attention, particularly under the influence of stigma in the Chinese societies.


Multi-Modal Curriculum Learning for Semi-Supervised Image Classification.
Semi-supervised image classification aims to classify a large quantity of unlabeled images by typically harnessing scarce labeled images. Existing semi-supervised methods often suffer from inadequate classification accuracy when encountering difficult yet critical images, such as outliers, because they treat all unlabeled images equally and conduct classifications in an imperfectly ordered sequence. In this paper, we employ the curriculum learning methodology by investigating the difficulty of classifying every unlabeled image. The reliability and the discriminability of these unlabeled images are particularly investigated for evaluating their difficulty. As a result, an optimized image sequence is generated during the iterative propagations, and the unlabeled images are logically classified from simple to difficult. Furthermore, since images are usually characterized by multiple visual feature descriptors, we associate each kind of features with a teacher, and design a multi-modal curriculum learning (MMCL) strategy to integrate the information from different feature modalities. In each propagation, each teacher analyzes the difficulties of the currently unlabeled images from its own modality viewpoint. A consensus is subsequently reached among all the teachers, determining the currently simplest images (i.e., a curriculum), which are to be reliably classified by the multi-modal learner. This well-organized propagation process leveraging multiple teachers and one learner enables our MMCL to outperform five state-of-the-art methods on eight popular image data sets.


High-density mapping suggests cytoplasmic male sterility with two restorer genes in almond × peach progenies.
Peach (Prunus persica) and almond (Prunus dulcis) are two sexually compatible species that produce fertile offspring. Almond, a highly polymorphic species, is a potential source of new genes for peach that has a strongly eroded gene pool. Here we describe the genetics of a male sterile phenotype that segregated in two almond ('Texas') × peach ('Earlygold') progenies: an F2 (T×E) and a backcross one (T1E) to the 'Earlygold' parent. High-density maps were developed using a 9k peach SNP chip and 135 simple-sequence repeats. Three highly syntenic and collinear maps were obtained: one for the F2 (T×E) and two for the backcross, T1E (for the hybrid) and E (for 'Earlygold'). A major reduction of recombination was observed in the interspecific maps (T×E and T1E) compared to the intraspecific parent (E). The E map also had extensive monomorphic genomic regions suggesting the presence of large DNA fragments identical by descent. Our data for the male sterility character were consistent with the existence of cytoplasmic male sterility, where individuals having the almond cytoplasm required the almond allele in at least one of two independent restorer genes, Rf1 and Rf2, to be fertile. The restorer genes were located in a 3.4 Mbp fragment of linkage group 2 (Rf1) and 1.4 Mbp of linkage group 6 (Rf2). Both fragments contained several genes coding for pentatricopeptide proteins, demonstrated to be responsible for restoring fertility in other species. The implications of these results for using almond as a source of novel variability in peach are discussed.


Innate Immune Recognition of EBV.
The ability of Epstein-Barr virus (EBV) to establish latency despite specific immune responses and to successfully persist lifelong in the human host shows that EBV has developed powerful strategies and mechanisms to exploit, evade, abolish, or downsize otherwise effective immune responses to ensure its own survival. This chapter focuses on current knowledge on innate immune responses against EBV and its evasion strategies for own benefit and summarizes the questions that remain to be tackled. Innate immune reactions against EBV originate both from the main target cells of EBV and from nontarget cells, which are elements of the innate immune system. Thus, we structured our review accordingly but with a particular focus on the innate recognition of EBV in its two stages in its life cycle, latent state and lytic replication. Specifically, we discuss (I) innate sensing and resulting innate immune responses against EBV by its main target cells, focusing on (i) EBV transmission between epithelial cells and B cells and their life cycle stages; and (ii) elements of innate immunity in EBV's target cells. Further, we debate (II) the innate recognition and resulting innate immune responses against EBV by cells other than the main target cells, focusing on (iii) myeloid cells: dendritic cells, monocytes, macrophages, and neutrophil granulocytes; and (iv) natural killer cells. Finally, we address (III) how EBV counteracts or exploits innate immunity in its latent and lytic life cycle stages, concentrating on (v) TLRs; (vi) EBERs; and (vii) microRNAs.


Sprouty genes control diastema tooth development via bidirectional antagonism of epithelial-mesenchymal FGF signaling.
Unlike humans, who have a continuous row of teeth, mice have only molars and incisors separated by a toothless region called a diastema. Although tooth buds form in the embryonic diastema, they regress and do not develop into teeth. Here, we identify members of the Sprouty (Spry) family, which encode negative feedback regulators of fibroblast growth factor (FGF) and other receptor tyrosine kinase signaling, as genes that repress diastema tooth development. We show that different Sprouty genes are deployed in different tissue compartments--Spry2 in epithelium and Spry4 in mesenchyme--to prevent diastema tooth formation. We provide genetic evidence that they function to ensure that diastema tooth buds are refractory to signaling via FGF ligands that are present in the region and thus prevent these buds from engaging in the FGF-mediated bidirectional signaling between epithelium and mesenchyme that normally sustains tooth development.


Combined bicarbonate conductance-impairing variants in CFTR and SPINK1 variants are associated with chronic pancreatitis in patients without cystic fibrosis.
Idiopathic chronic pancreatitis (ICP) is a complex inflammatory disorder associated with multiple genetic and environmental factors. In individuals without cystic fibrosis (CF), variants of CFTR that inhibit bicarbonate conductance but maintain chloride conductance might selectively impair secretion of pancreatic juice, leading to trypsin activation and pancreatitis. We investigated whether sequence variants in the gene encoding the pancreatic secretory trypsin inhibitor SPINK1 further increase the risk of pancreatitis in these patients. We screened patients and controls for variants in SPINK1 associated with risk of chronic pancreatitis and in all 27 exons of CFTR. The final study group included 53 patients with sporadic ICP, 27 probands with familial ICP, 150 unrelated controls, 375 additional controls for limited genotyping. CFTR wild-type and p.R75Q were cloned and expressed in HEK293 cells, and relative conductances of HCO(3)(-) and Cl(-) were measured. SPINK1 variants were identified in 36% of subjects and 3% of controls (odds ratio [OR], 18.1). One variant of CFTR not associated with CF, p.R75Q, was found in 16% of subjects and 5.3% of controls (OR, 3.4). Coinheritance of CFTR p.R75Q and SPINK1 variants occurred in 8.75% of patients and 0.38% of controls (OR, 25.1). Patch-clamp recordings of cells that expressed CFTR p.R75Q showed normal chloride currents but significantly reduced bicarbonate currents (P = .0001). The CFTR variant p.R75Q causes a selective defect in bicarbonate conductance and increases risk of pancreatitis. Coinheritance of p.R75Q or CF causing CFTR variants with SPINK1 variants significantly increases the risk of ICP.


Long-term treatment of hereditary angioedema with attenuated androgens: a survey of a 13-year experience.
Fifty-six patients affected with hereditary angioedema have been followed during long-term prophylaxis with attenuated androgens. The treatment was started in patients who had one or more severe attacks per month. In 24 patients, the therapy lasted for more than 5 years. The minimal effective dose usually did not exceed 2 mg/day of stanozolol or 200 mg/day of danazol. Only in two patients were these doses not sufficient to achieve the complete disappearance of symptoms. Irregular menstruation, but rarely amenorrhea, was the only significant side effect. One patient had to stop the therapy because of laboratory signs of hepatic cell necrosis. In one patient, danazol was administered during the last 8 weeks of pregnancy without side effects for the mother but with transient signs of virilization for the female baby. To find a biochemical marker for the minimal effective dose of androgen derivatives, we measured the plasma levels of C1 C1 INH complexes at different doses of stanozolol in four patients with hereditary angioedema. We found that these complexes, elevated before treatment, promptly reverted to normal values during androgen therapy and remained normal with any reduction of the dose of the drug as long as the patient remained symptom free. Therefore, the measurement of C1 C1 INH complexes appears to reflect the activity of the disease and not the amount of androgen that is administered.


A systematic method for clinical description and classification of personality variants. A proposal.
A systematic method for clinical description and classification of both normal and abnormal personality variants is proposed based on a general biosocial theory of personality. Three dimensions of personality are defined in terms of the basic stimulus-response characteristics of novelty seeking, harm avoidance, and reward dependence. The possible underlying genetic and neuroanatomical bases of observed variation in these dimensions are reviewed and considered in relation to adaptive responses to environmental challenge. The functional interaction of these dimensions leads to integrated patterns of differential response to novelty, punishment, and reward. The possible tridimensional combinations of extreme (high or low) variants on these basic stimulus-response characteristics correspond closely to traditional descriptions of personality disorders. This reconciles dimensional and categorical approaches to personality description. It also implies that the underlying structure of normal adaptive traits is the same as that of maladaptive personality traits, except for schizotypal and paranoid disorders.