Self-Assembly of Colloidal Molecules due to Self-Generated Flow.
The emergence of structure through aggregation is a fascinating topic and of both fundamental and practical interest. Here we demonstrate that self-generated solvent flow can be used to generate long-range attractions on the colloidal scale, with subpiconewton forces extending into the millimeter range. We observe a rich dynamic behavior with the formation and fusion of small clusters resembling molecules. The dynamics of this assembly is governed by an effective conservative energy that for large separations r decays as 1/r. Breaking the flow symmetry, these clusters can be made active.


Manifest latent nystagmus: a case of sensori-motor switching.
Latent nystagmus (LN) and manifest latent nystagmus (MLN) are closely associated with early visual deprivation and strabismus. In both cases, the eyes oscillate horizontally in an involuntary manner and the fast phases always beat towards the attending or fixing eye. By simultaneously recording eye movements during the dichoptic viewing of dissimilar visual stimuli we present evidence that MLN offers a unique opportunity to examine the nature of sensori-motor switching. In particular, we show how the nystagmus beat direction is strongly influenced by endogenous and exogenous attention. A model describing the possible mechanisms underpinning the sensori-motor switching is proposed.


Glial fibrillary acidic protein (GFAP) and the astrocyte intermediate filament system in diseases of the central nervous system.
Glial fibrillary acidic protein (GFAP) is the hallmark intermediate filament (IF; also known as nanofilament) protein in astrocytes, a main type of glial cells in the central nervous system (CNS). Astrocytes have a range of control and homeostatic functions in health and disease. Astrocytes assume a reactive phenotype in acute CNS trauma, ischemia, and in neurodegenerative diseases. This coincides with an upregulation and rearrangement of the IFs, which form a highly complex system composed of GFAP (10 isoforms), vimentin, synemin, and nestin. We begin to unravel the function of the IF system of astrocytes and in this review we discuss its role as an important crisis-command center coordinating cell responses in situations connected to cellular stress, which is a central component of many neurological diseases.


Can Some Marine-Derived Fungal Metabolites Become Actual Anticancer Agents?
Marine fungi are known to produce structurally unique secondary metabolites, and more than 1000 marine fungal-derived metabolites have already been reported. Despite the absence of marine fungal-derived metabolites in the current clinical pipeline, dozens of them have been classified as potential chemotherapy candidates because of their anticancer activity. Over the last decade, several comprehensive reviews have covered the potential anticancer activity of marine fungal-derived metabolites. However, these reviews consider the term "cytotoxicity" to be synonymous with "anticancer agent", which is not actually true. Indeed, a cytotoxic compound is by definition a poisonous compound. To become a potential anticancer agent, a cytotoxic compound must at least display (i) selectivity between normal and cancer cells (ii) activity against multidrug-resistant (MDR) cancer cells; and (iii) a preferentially non-apoptotic cell death mechanism, as it is now well known that a high proportion of cancer cells that resist chemotherapy are in fact apoptosis-resistant cancer cells against which pro-apoptotic drugs have more than limited efficacy. The present review thus focuses on the cytotoxic marine fungal-derived metabolites whose ability to kill cancer cells has been reported in the literature. Particular attention is paid to the compounds that kill cancer cells through non-apoptotic cell death mechanisms.


Functional analyses of epidemic Clostridioides difficile toxin B variants reveal their divergence in utilizing receptors and inducing pathology.
Clostridioides difficile toxin B (TcdB) is a key virulence factor that causes C. difficile associated diseases (CDAD) including diarrhea and pseudomembranous colitis. TcdB can be divided into multiple subtypes/variants based on their sequence variations, of which four (TcdB1-4) are dominant types found in major epidemic isolates. Here, we find that these variants are highly diverse in their receptor preference: TcdB1 uses two known receptors CSPG4 and Frizzled (FZD) proteins, TcdB2 selectively uses CSPG4, TcdB3 prefers to use FZDs, whereas TcdB4 uses neither CSPG4 nor FZDs. By creating chimeric toxins and systematically switching residues between TcdB1 and TcdB3, we determine that regions in the N-terminal cysteine protease domain (CPD) are involved in CSPG4-recognition. We further evaluate the pathological effects induced by TcdB1-4 with a mouse intrarectal installation model. TcdB1 leads to the most severe overall symptoms, followed by TcdB2 and TcdB3. When comparing the TcdB2 and TcdB3, TcdB2 causes stronger oedema while TcdB3 induces severer inflammatory cell infiltration. These findings together demonstrate divergence in the receptor preference and further lead to colonic pathology for predominant TcdB subtypes.


Should the dose of levothyroxine be changed in hypothyroidism patients fasting during Ramadan?
Muslims worship by fasting from predawn (suhoor) until sunset (iftar) for 30 days in the religious month of Ramadan. In addition to prolonged hunger, patients fasting with a diagnosis of hypothyroidism take their doses of levothyroxine (LT4) outside of daytime fasting hours. The purpose of our study is to compare the values of hypothyroid patients which have been obtained through thyroid function tests before and after Ramadan. Ninety-seven patients; ranging from 18 to 65 years old, who were followed with a diagnosis of hypothyroidism, who fasted during Ramadan, and who had no change of their LT4 dose for at least 6 months were included in the study. The median serum thyroid-stimulating hormone (TSH) level of patients prior to fasting was 2.19 mIU/L, while median serum TSH after fasting was 2.73 mIU/L. Serum TSH values after Ramadan increased significantly compared to those prior to Ramadan (P = 0.004). Our study demonstrates a significant increase in serum TSH levels after Ramadan but no significant change in serum free thyroxine (fT4) levels in hypothyroidism patients who are fasting. It may be appropriate to take precautions by making a small increase in LT4 dose before Ramadan in some hypothyroid patients wishing to fast.


Solubilized adenosine receptors in the brain: regulation of guanine nucleotides.
Adenosine receptors associated with a reduction of adenylate cyclase and labeled by tritium-labeled cyclohexyladenosine can be solubilized from brain membranes with sodium cholate. Regulation of receptor binding by guanine nucleotides is retained in the soluble state. Influences of cations observed in membrane preparations of adenosine receptors are no longer detected with the solubilized receptors. The apparent retention of a complex of receptors and guanosine triphosphate binding but not cation binding protein in the soluble state may permit a molecular analysis of receptor regulation.


NAVIGATE Program Directors' perspectives on treatment for early psychosis.
Since the release of the Recovery After Initial Schizophrenia Episode-Early Treatment Program results in 2015, the United States Congress expanded funding for early intervention programs and these programs now exist in every state. The purpose of the present study was to understand the real-world experience of NAVIGATE Program Directors with respect to identifying and recruiting patients with early psychosis and engaging families in treatment. Utilizing a mixed-methods sequential explanatory design, researchers surveyed 32 program directors in 13 states and engaged in 22 follow-up interviews to explore how NAVIGATE is being translated to serve individuals experiencing early psychosis and their families. We found that program directors using the NAVIGATE model encountered difficulty identifying diagnosis at admission and that supporting families of individuals experiencing psychosis was an important aspect of the program with unique complexity. These findings have important implications for future mental health research on early intervention programs and may serve to stimulate future research on how early intervention mental health programs can better serve individuals recovering from psychosis and support their families. (PsycInfo Database Record (c) 2023 APA, all rights reserved).


Circadian rhythms regulate amelogenesis.
Ameloblasts, the cells responsible for making enamel, modify their morphological features in response to specialized functions necessary for synchronized ameloblast differentiation and enamel formation. Secretory and maturation ameloblasts are characterized by the expression of stage-specific genes which follows strictly controlled repetitive patterns. Circadian rhythms are recognized as key regulators of the development and diseases of many tissues including bone. Our aim was to gain novel insights on the role of clock genes in enamel formation and to explore the potential links between circadian rhythms and amelogenesis. Our data shows definitive evidence that the main clock genes (Bmal1, Clock, Per1 and Per2) oscillate in ameloblasts at regular circadian (24 h) intervals both at RNA and protein levels. This study also reveals that the two markers of ameloblast differentiation i.e. amelogenin (Amelx; a marker of secretory stage ameloblasts) and kallikrein-related peptidase 4 (Klk4, a marker of maturation stage ameloblasts) are downstream targets of clock genes. Both, Amelx and Klk4 show 24h oscillatory expression patterns and their expression levels are up-regulated after Bmal1 over-expression in HAT-7 ameloblast cells. Taken together, these data suggest that both the secretory and the maturation stages of amelogenesis might be under circadian control. Changes in clock gene expression patterns might result in significant alterations of enamel apposition and mineralization.


Supplementation with Hualian No. 4 wild bitter gourd (Momordica charantia Linn. var. abbreviata ser.) extract increases anti-fatigue activities and enhances exercise performance in mice.
Hualian No. 4 wild bitter gourd (WBG) is a specific vegetable cultivated by the Hualien District Agricultural Research and Extension Station in Taiwan. WBG is commonly consumed as a vegetable and used as a popular folk medicine. However, few studies have demonstrated the effects of WBG supplementation on exercise performance, physical fatigue and the biochemical profile. The purpose of this study was to evaluate the potential beneficial effects of WBG extract on fatigue and ergogenic functions following physiological challenge. Three groups of male ICR mice (n=8 per group) were orally administered 0, 1 or 2.5 g/kg/day of WBG for 4 weeks. They were respectively designated the vehicle, WBG-1X and WBG-2.5X groups. WBG significantly decreased body weight (BW) and epididymal fat pad (EFP) weight. Concerning physical performance, WBG supplementation dose-dependently increased grip strength and endurance swimming time. Concerning anti-fatigue activity, WBG decreased levels of serum lactate, ammonia, creatine kinase and blood urea nitrogen, and economized glucose metabolism after acute exercise challenge. Glycogen in the liver and gastrocnemius muscle dose-dependently increased with WBG treatment. Concerning the biochemical profile, WBG treatment significantly decreased alanine aminotransferase (ALT), blood urea nitrogen (BUN) and urea acid (UA), and increased total protein (TP). Therefore, 4-week supplementation with WBG may decrease white adipose weight, enhance energy economy, increase glycogen storage to enhance exercise performance and reduce fatigue.


Self-Description of a University-Community Partnership to Advocate for Restroom Access Policy Change in Chicago.
Transgender, nonbinary, and gender-non-conforming individuals face significant obstacles accessing public restrooms, resulting in negative mental and physical health consequences. We describe the Chicago Restroom Access Project, a university and community partnership aimed at reforming restroom laws in Chicago and the state of Illinois. A range of community-engagement approaches were used, including recruiting, mobilizing, and partnering with diverse stakeholders, amplifying the voices of under-represented members of the impacted community, collecting data for evidence-based decision-making, fluid membership, and diffuse leadership. Outcomes included developing resources for public education, changing the City of Chicago Human Rights Ordinance, changing Illinois state law on single-occupancy restrooms, and implementing restroom reform at an educational institution. Lessons learned are also described. The methods and principles of this partnership provide approaches that can be used to advocate for reform and policy change for restroom access throughout the country.


Serovar distribution, antimicrobial resistance profiles, and PFGE typing of Salmonella enterica strains isolated from 2007-2012 in Guangdong, China.
Salmonella enterica includes the major serovars associated with human salmonellosis. In this study, 1764 clinical Salmonella enterica isolates from diarrhea outpatients were collected from fifteen cities in Guangdong province, China, between 2007 and 2012. These isolates represent all of the Salmonella isolates collected from the province during that period. The isolates were characterized by serovar determination, antimicrobial susceptibility tests and PFGE fingerprint typing. The serovar distribution results demonstrated that Salmonella Typhimurium (n=523, 29.65%) and Salmonella 4,5,12:i:- (n=244, 13.83%) are the most common serovars causing infant salmonellosis, whereas Salmonella Enteritidis (n=257, 14.57%) mainly causes human salmonellosis in adults. The serovar shift from Salmonella Enteritidis to Salmonella Typhimurium occurred in 2008. Antimicrobial susceptibility data showed a high burden of multidrug resistance (MDR) (n=1128, 56.58%), and a 20%-30% increase in the number of isolates resistant to ciprofloxacin (n=142, 8.05%) and third-generation cephalosporins (n=88, 4.99%) from 2007-2012. Only 9.97% of isolates (n=176) were fully susceptible to all agents tested. A high burden of MDR was observed in Salmonella Typhimurium and Salmonella 4,5,12:i:- for all age groups, and a reduced susceptibility to third-generation cephalosporins and quinolones occurred particularly in infants (≤ 6 years). The dominant PFGE patterns were JPXX01.GD0004, JEGX01.GD0006-7 and JNGX01.GD0006-7. ACSSuT was the predominant MDR profile in the Salmonella Typhimurium & 4,5,12:i:- complexes, while ASSuT-Nal and ASSu-Nal were the major MDR profiles in Salmonella Enteritidis. The predominant PFGE patterns of the Salmonella Typhimurium & 4,5,12:i:- complexes and Salmonella Stanley were most prevalent in infants (≤ 6 years). However, no obvious relationship was observed between these PFGE profiles and geographic location. These data reveal the serovar distribution of isolates recovered from diarrhea patients, the characteristics of resistant strains and fingerprint typing in Guangdong from 2007 to 2012. These results highlight a serovar shift and a worrying percentage of MDR strains with increasing resistance to quinolones and third-generation cephalosporins. Thus, continued surveillance of Salmonella and their MDR profiles using combined molecular tools and efforts to control the rapid increase in antimicrobial resistance among Salmonella in Guangdong are needed.


Viral etiologies of hospitalized acute lower respiratory infection patients in China, 2009-2013.
Acute lower respiratory infections (ALRIs) are an important cause of acute illnesses and mortality worldwide and in China. However, a large-scale study on the prevalence of viral infections across multiple provinces and seasons has not been previously reported from China. Here, we aimed to identify the viral etiologies associated with ALRIs from 22 Chinese provinces. Active surveillance for hospitalized ALRI patients in 108 sentinel hospitals in 24 provinces of China was conducted from January 2009-September 2013. We enrolled hospitalized all-age patients with ALRI, and collected respiratory specimens, blood or serum collected for diagnostic testing for respiratory syncytial virus (RSV), human influenza virus, adenoviruses (ADV), human parainfluenza virus (PIV), human metapneumovirus (hMPV), human coronavirus (hCoV) and human bocavirus (hBoV). We included 28,369 ALRI patients from 81 (of the 108) sentinel hospitals in 22 (of the 24) provinces, and 10,387 (36.6%) were positive for at least one etiology. The most frequently detected virus was RSV (9.9%), followed by influenza (6.6%), PIV (4.8%), ADV (3.4%), hBoV (1.9), hMPV (1.5%) and hCoV (1.4%). Co-detections were found in 7.2% of patients. RSV was the most common etiology (17.0%) in young children aged <2 years. Influenza viruses were the main cause of the ALRIs in adults and elderly. PIV, hBoV, hMPV and ADV infections were more frequent in children, while hCoV infection was distributed evenly in all-age. There were clear seasonal peaks for RSV, influenza, PIV, hBoV and hMPV infections. Our findings could serve as robust evidence for public health authorities in drawing up further plans to prevent and control ALRIs associated with viral pathogens. RSV is common in young children and prevention measures could have large public health impact. Influenza was most common in adults and influenza vaccination should be implemented on a wider scale in China.


High-χ alternating copolymers for accessing sub-5 nm domains via simulations.
The ever-growing semiconductor industry has encouraged the feature dimensions of nanolithography to reach the sub-10 nm length scale. It is highly necessary to find nanolithographic materials with high performance but ultra-small domains. We have designed a series of high-χ alternating copolymers (ACPs), in which the polar and apolar repeating units are four hydroxyl groups and alkyl chains, respectively. Careful coarse-grained molecular dynamics (CG-MD) simulations demonstrate that these ACPs can form a variety of mesophases, including lamellae, perforated lamellae, and hexagonally packed cylinders. All the domain periods of these mesophases are smaller than 5 nm, and the smallest domain is close to 1 nm. Most importantly, both the phase morphologies and domain periods are independent of the molecular weight (MW) and molecular weight distribution (MWD) when the degree of polymerization (N) exceeds the threshold value. Thus, using high-χ ACPs, ultranarrow domains can be realized with high MW for sufficient material performance, while the MWD-independence can ensure the uniformity of the domain sizes. We believe that these "high χ-high N" alternating copolymers are promising alternatives as new nanolithographic materials.


Fibroblast growth factor-23 in obese, normotensive adolescents is associated with adverse cardiac structure.
Fibroblast growth factor-23 (FGF23) is a biomarker for cardiovascular disease. Obesity may promote FGF23 production in the absence of chronic kidney disease. We sought to determine among normotensive African American adolescents whether FGF23 levels are greater in obese compared with normal-weight adolescents and to determine the relationship of FGF23 with markers of cardiac structure and insulin resistance. Cross-sectional data were obtained from a cohort of 130 normotensive, African American adolescents ages 13-18 years without chronic kidney disease; 74 were obese; 56 were normal weight. Plasma C-terminal FGF23, fasting glucose and insulin, and high-sensitivity C-reactive protein were measured; participants underwent M-mode echocardiography. FGF23 was skewed and approximately normally distributed after natural log transformation (logFGF23). FGF23 levels were greater in obese vs normal-weight participants (geometric mean 43 vs 23 RU/mL, P < .01). FGF23 values were significantly greater in participants with eccentric or concentric cardiac hypertrophy compared with those without hypertrophy P < .01). LogFGF23 directly correlated with body mass index, body mass index z-score, waist circumference, fasting insulin levels, and homeostasis model assessment scores. Regression models adjusted for age, sex, and high-sensitivity C-reactive protein suggest that each 10% increase in FGF23 is associated with a 1.31 unit increase in left ventricular mass (P < .01), a 0.29-unit increase in left ventricular mass index (P < .01), and a 0.01-unit increase in left atrial dimension indexed to height (P = .02). In this sample of obese African American adolescents, FGF23 blood levels were associated with abnormal cardiac structure. We postulate that FGF23 may be an early marker of cardiac injury in obese but otherwise-healthy African American adolescents.


Cleaning efficiency of anatomic endodontic technology, ProFile System and Manual Instrumentation in oval-shaped root canals: an in vitro study.
This in vitro study is an attempt to compare the effectiveness in cleaning oval shaped root canals using Anatomic Endodontic Technology (AET®), ProFile system® and Manual Instrumentation with K-files. Sixty oval shaped single rooted maxillary and mandibular premolars with straight canals were divided in to three groups. The root canals were, confirmed as being oval shape by means of radiographs made in a buccolingual and mesiodistal direction. Automated canal preparation was performed using Anatomic Endodontic Technology (group 1) and the ProFile system® (group 2). Manual instrumentation (group 3) was performed with k-files. Irrigation was performed using alternatively 3% NaOCl and 17% EDTA, followed by rinsing with normal saline. The roots were split longitudinally into two halves and examined under a scanning electron microscope. The presence of debris and smear layer was recorded at distances 1, 5 and 10 mm from the working length using a three step scoring scale. Mean scores for debris and smear layer was calculated and statistically analyzed for between and within groups significance, using the Kruskal-Wallis nonparametric ANOVA test and Bonferroni's multiple comparison test. At 1, 5 and 10 mm levels the root canals prepared with AET had significantly less surface debris and smear layer on the canal walls as compared to canals prepared with ProFile system® or manual instrumentation. For all three groups significantly lower mean smear layer scores (p < 0.05) were recorded at 5 and 10 mm levels compared with the 1 mm level. Significantly lower mean debris scores (p < 0.05) were also recorded at 5 and 10 mm levels for the AET group whereas no significant differences were found between the three levels for the ProFile system® and manual instrumentation groups. Although better instrumentation scores were obtained in canals prepared with AET, complete cleanliness was not achieved with any of the techniques and instruments investigated.


A pilot study of an online cognitive rehabilitation program for executive function skills in children with cancer-related brain injury.
Children with a history of cancer are at increased risk for cognitive impairments, particularly in executive and memory domains. Traditional, in-person cognitive rehabilitation strategies may be unavailable and/or impractical for many of these children given difficulties related to resources and health status. The feasibility and efficacy of implementing a computerized, home-based cognitive rehabilitation curriculum designed to improve executive function skills was examined in these children. A one-arm open trial pilot study of an original executive function cognitive rehabilitation curriculum was conducted with 23 paediatric cancer survivors aged 7-19. Compliance with the cognitive rehabilitation program was 83%, similar to that of many traditional programs. Following the cognitive intervention, participants showed significantly increased processing speed, cognitive flexibility, verbal and visual declarative memory scores as well as significantly increased pre-frontal cortex activation compared to baseline. These results suggest that a program of computerized cognitive exercises can be successfully implemented at home in young children with cancer. These exercises may be effective for improving executive and memory skills in this group, with concurrent changes in neurobiologic status.


Efficacy of vincristine and cyclophosphamide in the therapy of recurrent medulloblastoma.
We conducted a Phase II study of combination therapy with vincristine and cyclophosphamide in the treatment of patients with recurrent or metastatic medulloblastoma. Fourteen patients were treated with vincristine 2 mg/m2 (2.0-mg maximal dose) by intravenous bolus on Day 1 and cyclophosphamide 1 g/m2 by intravenous infusion on Days 1 and 2, with cycles repeated every 4 weeks. All 4 patients with extraneural disease (biopsy-proven bony metastases) responded (duration of responses 2+, 6+, 8, and 16+ months) and 4 of 8 evaluable patients with neuraxis disease responded (duration of response 2, 2+, 2+, and 21+ months). Toxicity was limited to neutropenia without any episodes of infection. These therapeutic results compare favorably with other reports of therapy for recurrent medulloblastoma and support the inclusion of vincristine and cyclophosphamide in randomized adjuvant therapy trials of patients with medulloblastoma.


Artificial Insemination as an Alternative Transmission Route for African Swine Fever Virus.
The rapid spread of the African swine fever virus (ASFV), causing severe disease with often high fatality rates in Eurasian suids, prevails as a threat for pig populations and dependent industries worldwide. Although advancing scientific progress continually enhances our understanding of ASFV pathogenesis, alternative transmission routes for ASFV have yet to be assessed. Here, we demonstrate that ASFV can efficiently be transferred from infected boars to naïve recipient gilts through artificial insemination (AI). In modern pig production, semen from boar studs often supplies many sow herds. Thus, the infection of a boar stud presents the risk of rapidly and widely distributing ASFV within or between countries. Daily blood and semen collection from four boars after intramuscular inoculation with ASFV strain 'Estonia 2014' resulted in the detection of ASFV genomes in the semen as early as 2 dpi, in blood at 1 dpi while semen quality remained largely unaffected. Ultimately, after insemination with extended semen, 7 of 14 gilts were ASFV positive by 7 days post insemination, and all gilts were ASFV positive by 35 days post insemination. Twelve out of 13 pregnant gilts aborted or resorbed at the onset of fever. A proportion of fetuses originating from the remaining gilt showed both abnormalities and replication of ASFV in fetal tissues. Thus, we present evidence for the efficient transmission of ASFV to gilts via AI and also to implanted embryos. These results underline the critical role that boar semen could play in ASFV transmission.


Exercises to prevent lower limb injuries in youth sports: cluster randomised controlled trial.
To investigate the effect of a structured warm-up programme designed to reduce the incidence of knee and ankle injuries in young people participating in sports. Cluster randomised controlled trial with clubs as the unit of randomisation. 120 team handball clubs from central and eastern Norway (61 clubs in the intervention group, 59 in the control group) followed for one league season (eight months). 1837 players aged 15-17 years; 958 players (808 female and 150 male) in the intervention group; 879 players (778 female and 101 male) in the control group. A structured warm-up programme to improve running, cutting, and landing technique as well as neuromuscular control, balance, and strength. The rate of acute injuries to the knee or ankle. During the season, 129 acute knee or ankle injuries occurred, 81 injuries in the control group (0.9 (SE 0.09) injuries per 1000 player hours; 0.3 (SE 0.17) in training v 5.3 (SE 0.06) during matches) and 48 injuries in the intervention group (0.5 (SE 0.11) injuries per 1000 player hours; 0.2 (SE 0.18) in training v 2.5 (SE 0.06) during matches). Fewer injured players were in the intervention group than in the control group (46 (4.8%) v (76 (8.6%); relative risk intervention group v control group 0.53, 95% confidence interval 0.35 to 0.81). A structured programme of warm-up exercises can prevent knee and ankle injuries in young people playing sports. Preventive training should therefore be introduced as an integral part of youth sports programmes.


Results of surgical treatment for thoracic myelopathy: minimum 2-year follow-up study in 132 patients.
Thoracic myelopathy is uncommon compared with cervical myelopathy. In this study, data obtained in patients with thoracic myelopathy caused by degenerative processes of the spine were retrospectively analyzed to clarify the surgical outcomes and to examine the various factors affecting the postoperative improvement. Between 1988 and 2002, 132 patients with thoracic myelopathy underwent surgery and a minimum 2-year observation period. Clinical data were collected from medical and operative records, and sagittal alignment of the spine was measured on radiographs. The patients were evaluated pre- and postoperatively using the modified Japanese Orthopaedic Association (JOA) scale (maximum score 11). The relationships among various factors affecting the preoperative severity of myelopathy and postoperative improvement were also examined. The population consisted of 97 men (mean age at surgery was 58 years) and 35 women (mean age at surgery 62 years). Myelopathy was caused by ossification of the ligamentum flavum (OLF) in 73 patients, ossification of the posterior longitudinal ligament (OPLL) in 21, combined OLF-OPLL in 10, intervertebral disc herniation (IDH) in 15, posterior bone spur in 11, and OLF with IDH or posterior bone spur in one patient each. The surgical outcome was relatively good: a mean preoperative JOA score of 5.3 improved to a mean score of 7.8 at the last follow-up, 50 months on average after surgery. Thoracic myelopathy caused by OPLL, however, was associated with lower postoperative scores and recovery rates. In more than half of the patients the authors documented an increase of kyphosis of less than 2 degrees. Patients with a shorter preoperative duration of symptoms and milder myelopathy experienced significantly better postoperative neurological conditions, which indicated that those who present earlier with fewer disabilities should be recommended to undergo surgery in time, although the surgical treatment for OPLL still involves many problems.


Blue fluorescent cGMP sensor for multiparameter fluorescence imaging.
Cyclic GMP (cGMP) regulates many physiological processes by cooperating with the other signaling molecules such as cyclic AMP (cAMP) and Ca(2+). Genetically encoded sensors for cGMP have been developed based on fluorescence resonance energy transfer (FRET) between fluorescent proteins. However, to analyze the dynamic relationship among these second messengers, combined use of existing sensors in a single cell is inadequate because of the significant spectral overlaps. A single wavelength indicator is an effective alternative to avoid this problem, but color variants of a single fluorescent protein-based biosensor are limited. In this study, to construct a new color fluorescent sensor, we converted the FRET-based sensor into a single wavelength indicator using a dark FRET acceptor. We developed a blue fluorescent cGMP biosensor, which is spectrally compatible with a FRET-based cAMP sensor using cyan and yellow fluorescent proteins (CFP/YFP). We cotransfected them and loaded a red fluorescent probe for Ca(2+) into cells, and accomplished triple-parameter fluorescence imaging of these cyclic nucleotides and Ca(2+), confirming the applicability of this combination to individually monitor their dynamics in a single cell. This blue fluorescent sensor and the approach using this FRET pair would be useful for multiparameter fluorescence imaging to understand complex signal transduction networks.


The role of voltage dependence of the NMDA receptor in cellular and network oscillation.
Unraveling the mechanisms underlying oscillatory behavior is critical for understanding normal and pathological brain processes. Here we used electrophysiology in mouse neocortical slices and principles of nonlinear dynamics to demonstrate how an increase in the N-methyl-d-aspartic acid receptor (NMDAR) conductance can create a nonlinear whole-cell current-voltage (I-V) relationship which leads to changes in cellular stability. We discovered two behaviorally and morphologically distinct pyramidal cell populations. Under control conditions, both cell types responded to depolarizing current injection with regular spiking patterns. However, upon NMDAR activation, an intrinsic oscillatory (IO) cell type (n = 44) showed a nonlinear whole-cell I-V relationship, intrinsic voltage-dependent oscillations plus amplification of alternating input current, and these properties persisted after disabling action potential generation with tetrodotoxin (TTX). The other non-oscillatory (NO) neuronal population (n = 24) demonstrated none of these behaviors. Simultaneous intra- and extracellular recordings demonstrated the NMDAR's capacity to promote low-frequency seizure-like network oscillations via its effects on intrinsic neuronal properties. The two pyramidal cell types demonstrated different relationships with network oscillation--the IO cells were leaders that were activated early in the population activity cycle while the activation of the NO cell type was distributed across network bursts. The properties of IO neurons disappeared in a low-magnesium environment where the voltage dependence of the receptor is abolished; concurrently, the cellular contribution to network oscillation switched to synchronous firing. Thus, depending upon the efficacy of NMDAR in altering the linearity of the whole-cell I-V relationship, the two cell populations played different roles in sustaining network oscillation.


International Latin American Survey on Pediatric Intestinal Failure Team.
There is little data on the experience of managing pediatric Intestinal Failure (IF) in Latin America. This study aimed to identify and describe the current organization and practices of the IF teams in Latin America and the Caribbean. An online survey was sent to inquire about the existence of IF teams that managed children on home parenteral nutrition (HPN). Our questionnaire was based on a previously published European study with a similar goal. Twenty-four centers with pediatric IF teams in eight countries completed the survey, representing a total number of 316 children on HPN. The median number of children on parenteral nutrition (PN) at home per team was 5.5 (range 1-50). Teams consisted of the following members: pediatric gastroenterologist and a pediatric surgeon in all teams, dietician (95.8%), nurse (91.7%), social worker (79.2%), pharmacist (70.8%), oral therapist (62.5%), psychologist (58.3%), and physiotherapist (45.8%). The majority of the centers followed international standards of care on vascular access, parenteral and enteral nutrition, and IF medical and surgical management, but a significant percentage reported inability to monitor micronutrients, like vitamins A (37.5%), E (41.7%), B1 (66.7%), B2 (62.5%), B6 (62.5%), active B12 (58.3%); and trace elements-including zinc (29.2%), aluminum (75%), copper (37.5%), chromium (58.3%), selenium (58.3%), and manganese (58.3%). Conclusion: There is wide variation in how IF teams are structured in Latin America-while many countries have well-established Intestinal rehabilitation programs, a few do not follow international standards. Many countries did not report having an IF team managing pediatric patients on HPN.


Macroamylasemia in HIV infection.
A case of macroamylasemia was seen in a 40-year-old HIV-positive bisexual male treated at the Fort Worth-Tarrant County Health Department (Ryan White Clinic). Macroamylasemia is a rare condition encountered sometimes in persons with HIV infection. Apart from the setting of HIV infection and acquired immunodeficiency syndrome, macroamylasemia is seen also in various conditions including liver disease, diabetes, cancer, malabsorption, and autoimmune disorders. Although this biochemical phenomenon requires no therapy, it should be considered in the differential diagnosis of patients who have persistently high levels of serum amylase and yet do not exhibit any clinical symptoms of pancreatitis or salivary gland inflammation.


The gene structure and promoter sequence of mouse hyaluronan synthase 1.
The structure and organization of mouse hyaluronan synthase 1 gene, HAS1 were determined by direct sequencing of lambda phage clones carrying the entire gene and by application of the long and accurate (LA)-PCR method to amplify regions encompassing the exon-intron boundaries and all of the exons. This gene spans about 11kb of genomic DNA and consists of 5 exons and 4 introns. A similarity in the exon-intron organization was found between the genes of mouse HAS1 and Xenopus laevis DG42 which was recently identified as Xenopus hyaluronan synthase. The transcription initiation site was determined by rapid amplification of the cDNA ends (5'-RACE). Position +1 is located 55 nucleotides upstream of the ATG initiation codon. The promoter region of the HAS1 gene has no typical TATA box, but contains a CCAAT box located 190 nucleotides upstream of the transcription initiation site. Further analysis of 1.4 kb of the 5' flanking region revealed several potential binding motifs for transcription factors. This information about the gene structure may be useful for further studies on the promoter activity.


[Pulmonary alveolar microlithiasis].
The literary data and own observation of pulmonary microlithiasis, an extremely rare disease, are presented. The results of investigation of local immunity in patients were depicted.


Comparative Y-chromosome analysis among Cypriots in the context of historical events and migrations.
Y-chromosome analysis provides valuable information regarding the migration patterns of male ancestors, ranging from the Paleolithic age to the modern humans. STR and SNP genotyping analysis provides data regarding the genetic and geographical ancestry of the populations studied. This study focused on the analysis of the Y-chromosome in Maronite Cypriots and Armenian Cypriots, who came to the island as a result of different historical events. The aim was to provide information on the paternal genetic ancestry of Maronites and Armenians of Cyprus and investigate any affinity with the Greek Cypriots and Turkish Cypriots of the island. Since there is limited information in the current literature, we proceeded and used 23 Y-chromosome STRs and 28 Y-chromosome SNPs to genotype 57 Maronite Cypriots and 56 Armenian Cypriots, which were then compared to data from 344 Greek Cypriots and 380 Turkish Cypriots. All samples were assigned to eight major Y-haplogroups but the most frequent haplogroup among all Cypriots is haplogroup J in the major subclade J2a-L559. The calculated pairwise genetic distances between the populations show that Armenian Cypriots are genetically closer to Greek and Turkish Cypriots compared to Maronite Cypriots. Median Joining Network analysis in 17 Y-STR haplotypes of all Cypriots assigned to J2a-L559, revealed that Cypriots share a common paternal ancestor, prior to the migration of the Armenians and Maronites to Cyprus, estimated in the Late Bronze Age and Early Iron Age.


The relation of the Halstead-Reitan neuropsychological battery to functional daily living skills in geriatric patients.
This study investigated the nature of the relationship between neuropsychological tests, the Halstead-Reitan Battery, and a measure of everyday living ability, the Community Competence Scale. The sample consisted of 40 geriatric patients referred for suspected dementia. The results indicated that neuropsychological tests were moderately predictive of measures of functional daily living skills. Although some specific neuropsychological measures, or combination of measures, were more predictive of particular functional daily living skills, their relationship did not necessarily follow logical or theoretical expectations. Thus, while it may be safe for neuropsychologists to make predictions about a patient's global capacity to function independently, predictions regarding specific functional daily living skills may be unwarranted.


Optimal intakes of protein in the human diet.
For protein, progress is slow in defining quantifiable indicators of adequacy other than balance and growth. As far as current requirements are concerned, only in the case of infants and children is there any case for revision, and this change is to lower values. Such intakes would appear to be safe when consumed as milk formula. In pregnancy, notwithstanding the concern that deficiency may influence programming of disease in later life, there is little evidence of any increased need, and some evidence that increased intakes would pose a risk. For the elderly there is no evidence of an increased requirement or of benefit from increased intakes, except possibly for bone health. For adults, while we now know much more about metabolic adaptation to varying intakes, there would appear to be no case for a change in current recommendations. As far as risks and benefits of high intakes are concerned, there is now only a weak case for risk for renal function. For bone health the established views of risk of high protein intakes are not supported by newly-emerging data, with benefit indicated in the elderly. There is also circumstantial evidence for benefit on blood pressure and stroke mortality. With athletes there is little evidence of benefit of increased intakes in terms of performance, with older literature suggesting an adverse influence. Thus, given that a safe upper limit is currently defined as twice the reference nutrient intake, and that for individuals with high energy requirements this value (1.5 g/kg per d) is easily exceeded, there is a case for revising the definition of a safe upper limit.


4D visualization of transitional structures in phase transformations by electron diffraction.
Complex systems in condensed phases involve a multidimensional energy landscape, and knowledge of transitional structures and separation of time scales for atomic movements is critical to understanding their dynamical behavior. Here, we report, using four-dimensional (4D) femtosecond electron diffraction, the visualization of transitional structures from the initial monoclinic to the final tetragonal phase in crystalline vanadium dioxide; the change was initiated by a near-infrared excitation. By revealing the spatiotemporal behavior from all observed Bragg diffractions in 3D, the femtosecond primary vanadium-vanadium bond dilation, the displacements of atoms in picoseconds, and the sound wave shear motion on hundreds of picoseconds were resolved, elucidating the nature of the structural pathways and the nonconcerted mechanism of the transformation.


Premotor cortex neurons in macaques: activity before distal and proximal forelimb movements.
Single unit activity was examined in the premotor cortex of three awake, behaving macaque monkeys. Premotor cortex (PM) neurons were found to be active in association with a movement involving specifically the distal forelimb. Other PM neurons were active in relation to more proximal forelimb movements. The spatial distribution of neurons related to distal movements differed from that of neurons related to proximal forelimb movements, the former being focused at a postarcuate region near the genu of the arcuate sulcus and the latter being shifted more posteromedially. The distal movement was triggered by one of the three sensory signals: visual, auditory, and vibrotactile. Of 190 PM neurons related to the distal forelimb movements, 123 (65%) showed similar premovement activity changes irrespective of the modality of the triggering signal. However, a number of neurons (35%) responded preferentially to one or two of the three signals. The modality-specific activity before movements distinguishes PM from the caudally adjacent primary motor cortex.


Survey of the use of intracranial pressure monitoring in children in the United Kingdom.
To establish current practice for the monitoring and management of acute intracranial hypertension in children in United Kingdom intensive care units (ICUs). Postal questionnaire, targetted by prior telephone survey, to all ICUs admitting five or more children per annum with acute neurological illness. Of the units contacted 70 % responded, approximately one-half of which reported the use of intracranial pressure (ICP) monitoring. Only data from these units are presented. Nearly all of these units consider monitoring following serious head injury, but its use in non-traumatic brain injury is less widespread. The decision to institute ICP monitoring is based mainly upon neuroimaging appearances and Glasgow Coma Scale score. ICP and cerebral perfusion pressure targets differ markedly between centres, with only 46 % and 65 % of units, respectively, setting age-dependent parameters. Mannitol and varying degrees of hyperventilation are employed by all units to lower ICP. The majority also use barbiturates, diuretics, and fluid restriction. Controlled hypothermia is used in 52 % of units. Paediatric units are more likely to employ age-dependent cerebral perfusion pressure targets. Specific therapies employed to lower ICP are similar to those used in adult centres. Faced with a lack of both evidence and consensus, the management of acute intracranial hypertension in childhood varies widely. National or international guidelines for the management of children with raised intracranial pressure are needed. These should incorporate the physiological differences between children of different ages.


Imaging pericytes and capillary diameter in brain slices and isolated retinae.
The cerebral circulation is highly specialized, both structurally and functionally, and it provides a fine-tuned supply of oxygen and nutrients to active regions of the brain. Our understanding of blood flow regulation by cerebral arterioles has evolved rapidly. Recent work has opened new avenues in microvascular research; for example, it has been demonstrated that contractile pericytes found on capillary walls induce capillary diameter changes in response to neurotransmitters, suggesting that pericytes could have a role in neurovascular coupling. This concept is at odds with traditional models of brain blood flow regulation, which assume that only arterioles control cerebral blood flow. The investigation of mechanisms underlying neurovascular coupling at the capillary level requires a range of approaches, which involve unique technical challenges. Here we provide detailed protocols for the successful physiological and immunohistochemical study of pericytes and capillaries in brain slices and isolated retinae, allowing investigators to probe the role of capillaries in neurovascular coupling. This protocol can be completed within 6-8 h; however, immunohistochemical experiments may take 3-6 d.


X-linked dominant hereditary motor and sensory neuropathy.
Modern techniques have defined the hereditary motor and sensory neuropathies (HMSN) as a genetically heterogeneous group of disorders. This includes a rare variant with X-linked dominant inheritance. We have traced this disorder through 6 generations of a large Canadian kindred; neurological and electrophysiological examinations were performed in 57 family members and nerve biopsies were studied in 2 affected males, early and late in the disease; 42/83 family members were affected. No male-to-male transmission was encountered in 19 sons of affected fathers, whereas all their daughters expressed the disease. Linkage was shown to the DNA loci DXYS1 Z max = 2.87 at theta max = 0.06 and to PGK1 Z max = 1.51 at theta max = 0 (Beckett et al., 1986). The typical clinical features are onset in early childhood, pes cavus, atrophy and weakness of peroneal muscles and intrinsic hand muscles, and sensory abnormalities. Males were severely affected, whereas females had mild or subclinical disease. Electrophysiological observations indicated a substantial loss of distal motor and sensory nerve fibres. Evoked compound muscle action potentials in extensor digitorum brevis were absent or severely reduced in 42% of cases and the peroneal motor nerve conduction velocity was mildly reduced to a mean 36.5 +/- 7.4 m.s-1. Sural sensory nerve action potentials were absent or severely reduced in 75% of those affected. Nerve biopsies showed loss of myelinated and unmyelinated nerve fibres, regenerative sprouting and secondary demyelination. The findings indicate that this distinct variant of HMSN is the result of primary axonal degeneration.


Distinct patterns of DNA copy number alteration are associated with different clinicopathological features and gene-expression subtypes of breast cancer.
Breast cancer is a leading cause of cancer-death among women, where the clinicopathological features of tumors are used to prognosticate and guide therapy. DNA copy number alterations (CNAs), which occur frequently in breast cancer and define key pathogenetic events, are also potentially useful prognostic or predictive factors. Here, we report a genome-wide array-based comparative genomic hybridization (array CGH) survey of CNAs in 89 breast tumors from a patient cohort with locally advanced disease. Statistical analysis links distinct cytoband loci harboring CNAs to specific clinicopathological parameters, including tumor grade, estrogen receptor status, presence of TP53 mutation, and overall survival. Notably, distinct spectra of CNAs also underlie the different subtypes of breast cancer recently defined by expression-profiling, implying these subtypes develop along distinct genetic pathways. In addition, higher numbers of gains/losses are associated with the "basal-like" tumor subtype, while high-level DNA amplification is more frequent in "luminal-B" subtype tumors, suggesting also that distinct mechanisms of genomic instability might underlie their pathogenesis. The identified CNAs may provide a basis for improved patient prognostication, as well as a starting point to define important genes to further our understanding of the pathobiology of breast cancer. This article contains Supplementary Material available at http://www.interscience.wiley.com/jpages/1045-2257/suppmat


Effects of 6-week sling-based training of the external-rotator muscles on the shoulder profile in elite female high school handball players.
In women's handball, the large numbers of throws and passes make the shoulder region vulnerable to overuse injuries. Repetitive throwing motions generate imbalance between shoulder internal- and external-rotator muscles. It has not yet been established whether sling-based training can improve shoulder external-rotator muscle strength. This study investigated the effectiveness of a 6-wk strengthening program in improving shoulder functional profile in elite female high school handball players. Crossover study. National elite handball training center. 25 elite female high school handball players. The program, completed twice per week for 6 wk, included sling-based strengthening exercises using a suspension trainer for external rotation with scapular retraction and scapular retraction alone. Maximal shoulder external- and internal-rotation strength, shoulder external- and internal-rotation range of motion (ROM), and maximal throwing velocity were assessed preintervention and postintervention for dominant and nondominant sides. After sling training, external- and internal-rotation strength increased significantly for both sides (P ≤ .001, and P = .004, respectively), with the result that there was no significant change in external- and internal-rotation strength ratios for either the dominant or the nondominant shoulder. No significant differences were observed for external-rotation ROM, while internal-rotation ROM decreased moderately, in particular in the dominant shoulder (P = .005). Maximal throwing velocity remained constant for the dominant arm, whereas a significant increase was found for the nondominant arm (P = .017). This 6-wk strengthening program was effective in improving shoulder external-rotator muscle strength but resulted in a decrease in the ROM in shoulder internal rotation, while throwing velocity remained stable. Adding a stretching program to this type of sling-based training program might help avoid potential detrimental effects on shoulder ROM.


Prognostic models for clinical outcomes in patients with venous leg ulcers: A systematic review.
The purpose of this review was to identify prognostic models for clinical application in patients with venous leg ulcers (VLUs). Literature searches were conducted in Embase, Medline, Cochrane, and CINAHL databases from inception to December 22, 2021. Eligible studies reported prognostic models aimed at developing, validating, and adjusting multivariable prognostic models that include multiple prognostic factors combined, and that predicted clinical outcomes. Methodological quality was assessed using the CHARMS checklist and PROBAST short form questionnaire. Thirteen studies were identified, of which three were validation studies of previously published models, four reported derivation and validation of models, and the remainder reported derivation models only. There was substantial heterogeneity in the model characteristics, including 11 studies focused on wound healing outcomes reporting 91 different predictors. Three studies shared similar predicted outcomes, follow-up timepoint and used a Cox proportional hazards model. However, these models reported different predictor selection methods and different predictors and it was therefore not feasible to summarize performance, such as discriminative ability. There are no standout risk prediction models in the literature with promising clinical application for patients with VLUs. Future research should focus on developing and validating high-performing models in wider VLU populations.


Characterization of two receptors for TRAIL.
Two receptors for TRAIL, designated TRAIL-R2 and TRAIL-R3, have been identified. Both are members of the tumor necrosis factor receptor family. TRAIL-R2 is structurally similar to the death-domain-containing receptor TRAIL-R1 (DR-4), and is capable of inducing apoptosis. In contrast, TRAIL-R3 does not promote cell death. TRAIL-R3 is highly glycosylated and is membrane bound via a putative phosphatidylinositol anchor. The extended structure of TRAIL-R3 is due to the presence of multiple threonine-, alanine-, proline- and glutamine-rich repeats (TAPE repeats). TRAIL-R2 shows a broad tissue distribution, whereas the expression of TRAIL-R3 is restricted to peripheral blood lymphocytes (PBLs) and skeletal muscle. All three TRAIL receptors bind TRAIL with similar affinity, suggesting a complex regulation of TRAIL-mediated signals.


pELMO, an optimised in-house cloning vector.
DNA cloning is an essential tool regarding DNA recombinant technology as it allows the replication of foreign DNA fragments within a cell. pELMO was here constructed as an in-house cloning vector for rapid and low-cost PCR product propagation; it is an optimally designed vector containing the ccdB killer gene from the pDONR 221 plasmid, cloned into the pUC18 vector's multiple cloning site (Thermo Scientific). The ccdB killer gene has a cleavage site (CCC/GGG) for the SmaI restriction enzyme which is used for vector linearisation and cloning blunt-ended products. pELMO transformation efficiency was evaluated with different sized inserts and its cloning efficiency was compared to that of the pGEM-T Easy commercial vector. The highest pELMO transformation efficiency was observed for ~500 bp DNA fragments; pELMO vector had higher cloning efficiency for all insert sizes tested. In-house and commercial vector cloned insert reads after sequencing were similar thus highlighting that sequencing primers were designed and localised appropriately. pELMO is thus proposed as a practical alternative for in-house cloning of PCR products in molecular biology laboratories.


Dosimetry Formalism and Implementation of a Homogenous Irradiation Protocol to Improve the Accuracy of Small Animal Whole-Body Irradiation Using a 137Cs Irradiator.
Shielded Cs irradiators are routinely used in pre-clinical radiation research to perform in vitro or in vivo investigations. Without appropriate dosimetry and irradiation protocols in place, there can be large uncertainty in the delivered dose of radiation between irradiated subjects that could lead to inaccurate and possibly misleading results. Here, a dosimetric evaluation of the JL Shepard Mark I-68A Cs irradiator and an irradiation technique for whole-body irradiation of small animals that allows one to limit the between subject variation in delivered dose to ±3% are provided. Mathematical simulation techniques and Gafchromic EBT film were used to describe the region within the irradiation cavity with homogeneous dose distribution (100% ± 5%), the dosimetric impact of varying source-to-subject distance, and the variation in attenuation thickness due to turntable rotation. Furthermore, an irradiation protocol and dosimetry formalism that allows calculation of irradiation time for whole-body irradiation of small animals is proposed that is designed to ensure a more consistent dose delivery between irradiated subjects. To compare this protocol with the conventional irradiation protocol suggested by the vendor, high-resolution film dosimetry measurements evaluating the dose difference between irradiation subjects and the dose distribution throughout subjects was performed using phantoms resembling small animals. Based on these results, there can be considerable variation in the delivered dose of > ± 5% using the conventional irradiation protocol for whole-body irradiation doses below 5 Gy. Using the proposed irradiation protocol this variability can be reduced to within ±3% and the dosimetry formalism allows for more accurate calculation of the irradiation time in relation to the intended prescription dose.


Evaluation of coagulation and fibrinolysis in horses with atrial fibrillation.
To evaluate horses with atrial fibrillation for hypercoagulability; plasma D-dimer concentrations, as a marker of a procoagulant state; and a relationship between coagulation profile results and duration of atrial fibrillation or presence of structural heart disease. Case-control study. Plasma samples from 42 horses (25 with atrial fibrillation and 17 without cardiovascular or systemic disease [control group]). Results of hematologic tests (ie, plasma fibrinogen and D-dimer concentrations, prothrombin and activated partial thromboplastin times, and antithrombin activity) in horses were recorded to assess coagulation and fibrinolysis. Historical and clinical variables, as associated with a hypercoagulable state in other species, were also recorded. Horses with atrial fibrillation and control horses lacked clinical signs of hypercoagulation or thromboembolism. Compared with control horses, horses with atrial fibrillation had significantly lower antithrombin activity. No significant differences in plasma fibrinogen and D-dimer concentrations and prothrombin and activated partial thromboplastin times existed between horse groups. In horses with atrial fibrillation versus control horses, a significantly larger proportion had an abnormal plasma D-dimer concentration (10/25 vs 2/17), test results indicative of subclinical activated coagulation (18/25 vs 6/17), or abnormal coagulation test results (25/121 vs 7/85), respectively. Horses with atrial fibrillation did not have clinical evidence of a hypercoagulable state, but a higher proportion of horses with atrial fibrillation, compared with control horses, did have subclinical activated coagulation on the basis of standard coagulation test results.


Lead poisoning secondary to hyperthyroidism: report of two cases.
With long-term exposure to lead, lead accumulates in bone, where it is stored for years. These quiescent lead stores are mobilised when increased bone turnover occurs, and latent lead toxicity may then become symptomatic. Although Graves' disease is a common cause of increased bone turnover, to date hyperthyroidism has been implicated in lead poisoning only twice. We describe herein two cases of hyperthyroidism, one caused by toxic multinodular thyroid enlargement, the second by Graves' disease, leading to lead poisoning. Treatment of hyperthyroidism with radioactive iodine cured both hyperthyroidism and lead poisoning and no chelating agent therapy was necessary. Lead poisoning is an important environmental health problem, and physicians must be aware of the endocrine disorders such as hyperthyroidism and hyperparathyroidism that increase bone turnover, favouring lead mobilisation. Atypical symptoms should draw the physician's attention to the possibility of lead poisoning, particularly in workers with occupational exposure to lead and in areas where lead poisoning is endemic.


Lack of sequence variation of the mitochondrial cytochrome b gene from a malaria parasite, Plasmodium mexicanum.
Very slight sequence differences in the mitochondrial cytochrome b gene, even single nucleotide substitutions, have been proposed as indicative of different species of avian malaria parasites. However, few studies have examined within-species variation in that gene for Plasmodium or related genera. We examined sequences for the entire cytochrome b gene from Plasmodium mexicanum , a parasite of lizards, for sites where microsatellite markers revealed substantial genetic diversity. For sites where the parasite is geographically genetically differentiated, and may have been isolated for thousands of years, there was no sequence variation (1,153 nucleotides) for >160 infections studied. The low degree of variation found in the cytochrome b gene for two human malaria parasites world-wide, as well as the lack of variation for P. mexicanum , contrast with the substantial variation found in surveys of bird malaria parasites, even in restricted geographic regions.


When Knowledge and Experience Do Not Help: A Study of Nonfatal Drug Overdoses.
: With recent increases in overdose deaths in Australia, there is renewed interest in understanding the factors that contribute to overdose. We examine the experiences of persons who report a nonfatal drug overdose. Fifty people who inject drugs (PWID) and who had accidently overdosed in the past 12 months were recruited and interviewed at 1 of 4 Needle and Syringe Program sites during September and October 2013.Participants were typically male, middle-aged, with long injecting histories. Half of the participants reported mainly injecting pharmaceutical opioids. Most overdoses occurred at home with others present. An ambulance was called for only 38% of cases and 26% were admitted to a hospital emergency department. Police were seldom involved, and there were no complaints about the involvement of police at the time of the overdose.Participants commonly had a history of overdosing, and most were on prescription medications for physical and/or mental health problems. Poly drug use was common for those reporting an accidental overdose. Benzodiazepines (eg, Xanax or Valium) were implicated in just over half of the overdoses.Most of those reporting a recent overdose also report a past history of previous overdoses. Most of those reporting a previous overdose continue to use substances in ways they are aware contribute to the risk of an overdose.


Validation of equations to estimate kidney function with and without adjustment by race/color in Brazilian adults (ELSA-Brazil).
To evaluate accuracy and agreement between creatinine clearance (CrCl) measured in 12-h urine and glomerular filtration rate (GFR) calculated by the Modification of Diet in Renal Disease (MDRD-4) and the Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) formulas, with and without adjustment for race/color. Baseline data from the Longitudinal Study of Adult Health (ELSA-Brazil) in adults (35-74 years of age) of both genders were used. Serum creatinine was measured in fasting blood and urinary creatinine was measured in an overnight 12-h urine collect. The agreement between CrCl and the calculated GFR was analyzed by the Bland-Altman method. One-way analysis of variance (ANOVA) with race/color factor was used to verify differences between means of CrCl and GFR with and without correction for race/color. Statistical significance was accepted for p<0.05. From 15,105 participants in the ELSA-Brazil, 12,813 had a validated urine collect. The Bland-Altman diagrams showed that formulas and CrCl agree with each other with a better accuracy for GFR <90 mL/.min x 1.73m2. The adjustment by race/color increased data dispersion. In this range, one-way ANOVA of CrCl with race/color factor showed similarity between groups (p=0.27). MDRD-4 and CKD-EPI are useful formulas for screening cases of chronic kidney disease, and correction by race/color, only in blacks or in black and brown subjects, proved to be unnecessary and reduced the reliability of the equations.


Cloning, expression, and characterization of the squid Na+-Ca2+ exchanger (NCX-SQ1).
We have cloned the squid neuronal Na+-Ca2+ exchanger, NCX-SQ1, expressed it in Xenopus oocytes, and characterized its regulatory and ion transport properties in giant excised membrane patches. The squid exchanger shows 58% identity with the canine Na+-Ca2+ exchanger (NCX1.1). Regions determined to be of functional importance in NCX1 are well conserved. Unique among exchanger sequences to date, NCX-SQ1 has a potential protein kinase C phosphorylation site (threonine 184) between transmembrane segments 3 and 4 and a tyrosine kinase site in the Ca2+ binding region (tyrosine 462). There is a deletion of 47 amino acids in the large intracellular loop of NCX-SQ1 in comparison with NCX1. Similar to NCX1, expression of NCX-SQ1 in Xenopus oocytes induced cytoplasmic Na+-dependent 45Ca2+ uptake; the uptake was inhibited by injection of Ca2+ chelators. In giant excised membrane patches, the NCX-SQ1 outward exchange current showed Na+-dependent inactivation, secondary activation by cytoplasmic Ca2+, and activation by chymotrypsin. The NCX-SQ1 exchange current was strongly stimulated by both ATP and the ATP-thioester, ATP gamma S, in the presence of F- (0.2 mM) and vanadate (50 microM), and both effects reversed on application of a phosphatidylinositol-4',5'-bisphosphate antibody. NCX1 current was stimulated by ATP, but not by ATP gamma S. Like NCX1 current, NCX-SQ1 current was strongly stimulated by phosphatidylinositol-4',5'-bisphosphate liposomes. In contrast to results in squid axon, NCX-SQ1 was not stimulated by phosphoarginine (5-10 mM). After chymotrypsin treatment, both the outward and inward NCX-SQ1 exchange currents were more strongly voltage dependent than NCX1 currents. Ion concentration jump experiments were performed to estimate the relative electrogenicity of Na+ and Ca2+ transport reactions. Outward current transients associated with Na+ extrusion were much smaller for NCX-SQ1 than NCX1, and inward current transients associated with Ca2+ extrusion were much larger. For NCX-SQ1, charge movements of Ca2+ transport could be defined in voltage jump experiments with a low cytoplasmic Ca2+ (2 microM) in the presence of high extracellular Ca2+ (4 mM). The rates of charge movements showed "U"-shaped dependence on voltage, and the slopes of both charge-voltage and rate-voltage relations (1,600 s-1 at 0 mV) indicated an apparent valency of -0.6 charges for the underlying reaction. Evidently, more negative charge moves into the membrane field in NCX-SQ1 than in NCX1 when ions are occluded into binding sites.


A peculiar sequence motif in the alpha-1-domain of the HLA-C molecule in psoriasis.
Most immunogenetic studies restrict the location of the susceptibility gene for psoriasis to the HLA-C locus. The present study evaluates the amino acid sequences of the HLA-C binding groove associated with the disease in a restricted population of Italian descent: Pavia, Northern Italy. Forty patients, affected by psoriasis, underwent genomic analysis of exons 2 and 3 of the HLA-C gene by means of the PCR-SSP technique. The HLA-C*0602 gene frequency was significantly higher in patients with respect to 122 sex-, age- and ethnically matched controls (25 vs. 8.6%, chi(2) = 14.65, p = 0.000129). This frequency rose to 39.5% in patients with a family history positive for psoriasis. In particular, 4 amino acid substitutions were shown to be crucial for the predisposition to the disease, namely Ala 73 and Asn 77 (located in the C pocket of the antigen binding groove), Lys 80 in the F pocket and Asp 90 in the outer loop of the molecule. This peculiar motif, entirely located in the alpha(1)-domain of the HLA-C molecule, was characteristic of psoriatic patients being present in 77.5% of the cases against the 50.82% of controls (chi(2) = 8.77; p = 0.0031). Homozygosity for the amino acids considered at these residues was found in 15% of patients and in 0% of controls (chi(2) = 20.74, p = 0.0000053). The most important results emerging from this study indicate that Ala 73, Asn 77, Lys 80 and Asp 90 together form a motif restricted to few pathogenic peptides to be studied in the near future.


Synergistic role of curli and cellulose in cell adherence and biofilm formation of attaching and effacing Escherichia coli and identification of Fis as a negative regulator of curli.
Curli are adhesive fimbriae of Escherichia coli and Salmonella enterica. Expression of curli (csgA) and cellulose (bcsA) is co-activated by the transcriptional activator CsgD. In this study, we investigated the contribution of curli and cellulose to the adhesive properties of enterohaemorragic (EHEC) O157:H7 and enteropathogenic E. coli (EPEC) O127:H6. While single mutations in csgA, csgD or bcsA in EPEC and EHEC had no dramatic effect on cell adherence, double csgAbcsA mutants were significantly less adherent than the single mutants or wild-type strains to human colonic HT-29 epithelial cells or to cow colon tissue in vitro. Overexpression of csgD (carried on plasmid pCP994) in a csgD mutant, but not in the single csgA or bscA mutants, led to significant increase in adherence and biofilm formation in EPEC and EHEC, suggesting that synchronized over-production of curli and cellulose enhances bacterial adherence. In line with this finding, csgD transcription was activated significantly in the presence of cultured epithelial cells as compared with growth in tissue culture medium. Analysis of the influence of virulence and global regulators in the production of curli in EPEC identified Fis (factor for inversion stimulation) as a, heretofore unrecognized, negative transcriptional regulator of csgA expression. An EPEC E2348/69Deltafis produced abundant amounts of curli whereas a double fis/csgD mutant yielded no detectable curli production. Our data suggest that curli and cellulose act in concert to favour host colonization, biofilm formation and survival in different environments.


Neural tube and craniofacial defects with special emphasis on folate pathway genes.
Neural tube and orofacial defects are common congenital malformations in humans. While etiologically heterogeneous, they are for the most part multifactorial in their pathogenesis, having both genetic and environmental components in their development. In recent years, there has been a great deal of epidemiologic evidence demonstrating that women who received multivitamins containing folic acid periconceptionally had significantly reduced occurrence and recurrence risks for producing infants with such malformations. This risk reduction is not observed in all populations, further suggestive of a genetic regulation of this phenomenon. Unfortunately, the mechanisms underlying the beneficial effects of folic acid are not well-understood. In this article, we review the relevant epidemiologic data on both neural tube defects and orofacial malformations, the fundamental embryological processes involved in closing the neural tube, and the development of the craniofacies, and propose a working hypothesis for susceptibility to these malformations. This hypothesis is based on the interworkings of cellular folate transport, focusing on the key elements involved in potocytosis. We propose that infants with mutations in the folate receptor alpha gene might be at increased risk for congenital anomalies due to a reduced binding affinity for 5-methyltetrahydrofolate, the physiologic form of folic acid. Various experimental approaches to test the working hypothesis are considered.


Current concepts in vital primary pulp therapy.
Recent progress in understanding the molecular and cellular changes during tooth development and how they are mimicked during tissue repair, offers the opportunity to assess the biologic validity of the various vital pulp treatments. Under this light, indirect pulp treatment can be an acceptable procedure for primary teeth with reversible pulp inflammation, provided that this diagnosis is based on a good history, a proper clinical and radiographic examination, and the tooth had been sealed with a leakage-free restoration. Several articles report the success of this technique of direct pulp capping (DPC) and calcium hydroxide has been widely used with high success rates in young permanent teeth, but the results in primary teeth are less satisfactory. Recent studies have reported successful results with direct adhesive capping of exposed pulps, while others showed pulp inflammation and unacceptable results using this technique. Thus, the traditional rationale for the use of calcium hydroxide should be maintained, and this treatment modality reserved for iatrogenic exposures in asymptomatic teeth that are expected to exfoliate within a short period of time. In younger children, iatrogenic or carious exposures should be treated by pulpotomy. Formocresol has been the most popular pulp dressing material for pulpotomized primary molars for many years but, due to its deleterious effect, the use of formocresol is decreasing considerably worldwide. Ferric sulphate has been proposed as a substitute to formocresol, and the success rates were comparable to those of formocresol. More recently, considerably better results have been obtained with MTA (Mineral Trioxide Aggregate), and statistically significant differences were reported when compared with formocresol. Internal root resorption, a finding seen both in ferric sulphate and formocresol, was not observed in the MTA treated teeth. MTA is commercially available, but its cost is very high, and cannot be kept once opened. Thus, ferric sulphate can still be a valid and inexpensive solution for pulpotomies in primary teeth.


Generation and validation of a conditional knockout mouse model for the study of the Smith-Lemli-Opitz syndrome.
Smith-Lemli-Opitz Syndrome (SLOS) is a developmental disorder (OMIM #270400) caused by autosomal recessive mutations in the Dhcr7 gene, which encodes the enzyme 3β-hydroxysterol-Δ7 reductase. SLOS patients present clinically with dysmorphology and neurological, behavioral, and cognitive defects, with characteristically elevated levels of 7-dehydrocholesterol (7-DHC) in all bodily tissues and fluids. Previous mouse models of SLOS have been hampered by postnatal lethality when Dhcr7 is knocked out globally, while a hypomorphic mouse model showed improvement in the biochemical phenotype with aging and did not manifest most other characteristic features of SLOS. We report the generation of a conditional knockout of Dhcr7 (Dhcr7flx/flx), validated by generating a mouse with a liver-specific deletion (Dhcr7L-KO). Phenotypic characterization of liver-specific knockout mice revealed no significant changes in viability, fertility, growth curves, liver architecture, hepatic triglyceride secretion, or parameters of systemic glucose homeostasis. Furthermore, qPCR and RNA-Seq analyses of livers revealed no perturbations in pathways responsible for cholesterol synthesis, either in male or in female Dhcr7L-KO mice, suggesting that hepatic disruption of postsqualene cholesterol synthesis leads to minimal impact on sterol metabolism in the liver. This validated conditional Dhcr7 knockout model may now allow us to systematically explore the pathophysiology of SLOS, by allowing for temporal, cell and tissue-specific loss of DHCR7.


Technology and Information Sharing in Disaster Relief.
This paper seeks to examine the extent to which technological advances can enhance inter-organizational information sharing in disaster relief. Our case is the Virtual OSOCC (On-Site Operations Coordination Centre) which is a part of the Global Disaster Alert and Coordination System (GDACS) under the United Nations Office for Coordination of Humanitarian Affairs (UN OCHA). The online platform, which has been developing for more than a decade, provides a unique insight into coordination behaviour among disaster management agencies and individual actors. We build our study on the analysis of a complete database of user interaction including more than 20,000 users and 11,000 comments spread across approximately 300 disaster events. Controlling for types and severities of the events, location-specific vulnerabilities, and the overall trends, we find that the introduction of new features have led to increases in user activity. We supplement the data-driven approach with evidence from semi-structured interviews with administrators and key users, as well as a survey among all users specifically designed to capture and assess the elements highlighted by both interviews and data analysis.


Spatially valid proprioceptive cues improve the detection of a visual stimulus.
Vision and proprioception are the main sensory modalities that convey hand location and direction of movement. Fusion of these sensory signals into a single robust percept is now well documented. However, it is not known whether these modalities also interact in the spatial allocation of attention, which has been demonstrated for other modality pairings. The aim of this study was to test whether proprioceptive signals can spatially cue a visual target to improve its detection. Participants were instructed to use a planar manipulandum in a forward reaching action and determine during this movement whether a near-threshold visual target appeared at either of two lateral positions. The target presentation was followed by a masking stimulus, which made its possible location unambiguous, but not its presence. Proprioceptive cues were given by applying a brief lateral force to the participant's arm, either in the same direction (validly cued) or in the opposite direction (invalidly cued) to the on-screen location of the mask. The d' detection rate of the target increased when the direction of proprioceptive stimulus was compatible with the location of the visual target compared to when it was incompatible. These results suggest that proprioception influences the allocation of attention in visual space.


Case Series: Keratolimbal Allograft as a Patch Graft for Glaucoma Drainage Devices.
Tube exposure remains one of the most common complications after glaucoma drainage device (GDD) implantation, despite various types of patch grafts available today. We present a 4 patient case series following the effectivity of the keratolimbal allograft (KLAL) as a patch graft for cases of tube exposure. Given its inherent population of stem cells, our hypothesis was that this highly replicative, biological tissue would provide an adequate means of glaucoma tube coverage. The subset of patients chosen for the KLAL patch graft all had a history of abnormally scarred conjunctiva or thin sclera. The aim of utilizing the KLAL patch with its associated donor conjunctival and scleral ring was also to provide additional reinforcement and adequate tube coverage in the setting of compromised native tissue. Four patients comprised of 2 males and 2 females with a minimum postoperative period of 12 months. All GDDs were initially implanted with a limbal-based incision using either Ahmed glaucoma valve or Baerveldt drainage implant. Three of the 4 patients received the KLAL patch graft after tube exposure with scleral patch graft and 1 patient received KLAL as the primary graft during initial tube placement. Two of the 4 eyes experienced tube re-exposure postoperatively at 2 and 3 months, respectively. Both of these cases had a history of prior tube exposure after scleral patch graft and both were tubes placed in the pars plana. Interestingly, the patients with failed grafts were younger with a history of more ocular surgeries as compared with the patients with graft viable eyes. Through our case series, we found that the KLAL utilized as a patch graft over GDD tubes has the potential for favorable outcomes in certain subtypes of eyes. Although further large scale investigation will be necessary to better define the risk factors associated with graft failure, proving the graft's viability is a crucial first step.


Chronic sustained and intermittent hypoxia reduce function of ATP-sensitive potassium channels in nucleus of the solitary tract.
Activation of neuronal ATP-sensitive potassium (K(ATP)) channels is an important mechanism that protects neurons and conserves neural function during hypoxia. We investigated hypoxia (bath gassed with 95% N(2)-5% CO(2) vs. 95% O(2)-5% CO(2) in control)-induced changes in K(ATP) current in second-order neurons of peripheral chemoreceptors in the nucleus of the solitary tract (NTS). Hypoxia-induced K(ATP) currents were compared between normoxic (Norm) rats and rats exposed to 1 wk of either chronic sustained hypoxia (CSH) or chronic intermittent hypoxia (CIH). Whole cell recordings of NTS second-order neurons identified after 4-(4-(dihexadecylamino)styryl)-N-methylpyridinium iodide (DiA) labeling of the carotid bodies were obtained in a brain stem slice. In Norm cells (n = 9), hypoxia (3 min) induced an outward current of 12.7 +/- 1.1 pA with a reversal potential of -73 +/- 2 mV. This current was completely blocked by the K(ATP) channel blocker tolbutamide (100 muM). Bath application of the K(ATP) channel opener diazoxide (200 muM, 3 min) evoked an outward current of 21.8 +/- 5.8 pA (n = 6). Hypoxia elicited a significantly smaller outward current in both CSH (5.9 +/- 1.4 pA, n = 11; P < 0.01) and CIH (6.8 +/- 1.7 pA, n = 6; P < 0.05) neurons. Diazoxide elicited a significantly smaller outward current in CSH (3.9 +/- 1.0 pA, n = 5; P < 0.05) and CIH (2.9 +/- 0.9 pA, n = 3; P < 0.05) neurons. Western blot analysis showed reduced levels of K(ATP) potassium channel subunits Kir6.1 and Kir6.2 in the NTS from CSH and CIH rats. These results suggest that hypoxia activates K(ATP) channels in NTS neurons receiving monosynaptic chemoreceptor afferent inputs. Chronic exposure to either sustained or intermittent hypoxia reduces K(ATP) channel function in NTS neurons. This may represent a neuronal adaptation that preserves neuronal excitability in crucial relay neurons in peripheral chemoreflex pathways.


Language processing in the occipital cortex of congenitally blind adults.
Humans are thought to have evolved brain regions in the left frontal and temporal cortex that are uniquely capable of language processing. However, congenitally blind individuals also activate the visual cortex in some verbal tasks. We provide evidence that this visual cortex activity in fact reflects language processing. We find that in congenitally blind individuals, the left visual cortex behaves similarly to classic language regions: (i) BOLD signal is higher during sentence comprehension than during linguistically degraded control conditions that are more difficult; (ii) BOLD signal is modulated by phonological information, lexical semantic information, and sentence-level combinatorial structure; and (iii) functional connectivity with language regions in the left prefrontal cortex and thalamus are increased relative to sighted individuals. We conclude that brain regions that are thought to have evolved for vision can take on language processing as a result of early experience. Innate microcircuit properties are not necessary for a brain region to become involved in language processing.


Structural dynamics of GluK2 kainate receptors in apo and partial agonist bound states.
Kainate receptors (KARs) belong to the family of ionotropic glutamate receptors (iGluRs) and are tetrameric ligand-gated ion channels that regulate neurotransmitter release and excitatory synaptic transmission in the central nervous system. While KARs share overall architectures with other iGluR subfamilies, their dynamics are significantly different from those of other iGluRs. KARs are activated by both full and partial agonists. While there is less efficacy with partial agonists than with full agonists, the detailed mechanism has remained elusive. Here, we used cryo-electron microscopy to determine the structures of homomeric rat GluK2 KARs in the absence of ligands (apo) and in complex with a partial agonist. Intriguingly, the apo state KARs were captured in desensitized conformation. This structure confirms the KAR desensitization prior to activation. Structures of KARs complexed to the partial agonist domoate populate in domoate bound desensitized and non-active/non-desensitized states. These previously unseen intermediate structures highlight the molecular mechanism of partial agonism in KARs. Additionally, we show how N -glycans stabilized the ligand-binding domain dimer via cation/anion binding and modulated receptor gating properties using electrophysiology. Our findings provide vital structural and functional insights into the unique KAR gating mechanisms.


[Vesico-ureteral reflux associated with detrusor instability].
The aim of this study was to determine the urodynamic characteristics of patients with vesico-ureteral reflux (VUR) and detrusor instability (DI) followed by medical treatment. The urodynamic and cystographic findings in a group of 24 patients between 4 and 18 years of age (mean 7.6 years) with a simultaneous diagnosis of VUR and DI were reviewed. All of them presented with recurrent urinary tract infections. Twenty were female of whom eight also had enuresis and daytime symptoms. Mean follow-up was 40 months (range 18-97 months). VUR was diagnosed by voiding cystourethrogram and classified according to the grades proposed by the "International reflux study on children". All but 6 patients had at least a 15% reduction in age-adjusted capacity. No relationship was observed between the severity of VUR (measured as the addition of reflux grades in both units of each patient) and reduction in bladder capacity or compliance. Seven patients had upper tract damage (either scars or a decrease in size or function on the renogram). Upper tract damage was significantly more frequent in patients with bilateral reflux (regardless of the type of DI). Patients with unilateral reflux and sustained instability had significantly less upper tract damage. Results of treatment are reported separately. In these patients, the severity of VUR did not seem to have an impact on bladder capacity or compliance. The risk factors for upper tract damage in this series differ from those found by other authors.


Cercosporamide inhibits bone morphogenetic protein receptor type I kinase activity in zebrafish.
Zebrafish models are well-established tools for investigating the underlying mechanisms of diseases. Here, we identified cercosporamide, a metabolite from the fungus Ascochyta aquiliqiae, as a potent bone morphogenetic protein receptor (BMPR) type I kinase inhibitor through a zebrafish embryo phenotypic screen. The developmental defects in zebrafish, including lack of the ventral fin, induced by cercosporamide were strikingly similar to the phenotypes caused by renowned small-molecule BMPR type I kinase inhibitors and inactivating mutations in zebrafish BMPRs. In mammalian cell-based assays, cercosporamide blocked BMP/SMAD-dependent transcriptional reporter activity and BMP-induced SMAD1/5-phosphorylation. Biochemical assays with a panel of purified recombinant kinases demonstrated that cercosporamide directly inhibited kinase activity of type I BMPRs [also called activin receptor-like kinases (ALKs)]. In mammalian cells, cercosporamide selectively inhibited constitutively active BMPR type I-induced SMAD1/5 phosphorylation. Importantly, cercosporamide rescued the developmental defects caused by constitutively active Alk2 in zebrafish embryos. We believe that cercosporamide could be the first of a new class of molecules with potential to be developed further for clinical use against diseases that are causally linked to overactivation of BMPR signaling, including fibrodysplasia ossificans progressiva and diffuse intrinsic pontine glioma.This article has an associated First Person interview with the first author of the paper.


The role of negative charge in spontaneous aggregation of transformed and untransformed cell lines.
No correlation between net negative surface charge as determined by electrophoretic light-scattering techniques and the rates of spontaneous aggregation of 3T3MIT and SVPy 3T3MIT cells has been found. Neuraminidase treatment of both 3T3MIT cells and SVPy 3T3MIT cells causes a significant decrease in electrophoretic mobility but only the 3T3MIT cells show an increase in spontaneous aggregation. An increase in spontaneous aggregation of 3T3MIT cells is seen after growth in 200 mM urea for 18 h but no change in net surface charge occurs. The distribution of anionic sites on the membranes of cells was determined using the ultrastructural marker polycationized ferritin. The distribution of polycationized ferritin-binding sites was essentially identical for both cell lines under all conditions when they were labelled at 4 degrees C. When the cells were labelled with polycationized ferritin at 37 degrees C it was found that cells which have a high net rate of spontaneous aggregation also show rearrangement of anionic sites on their surface membrane. Clustering and rearrangement of anionic sites at 37 degrees C correlate with high rates of spontaneous aggregation.


Transport and public health in China: the road to a healthy future.
Transportation-related risk factors are a major source of morbidity and mortality in China, where the expansion of road networks and surges in personal vehicle ownership are having profound effects on public health. Road traffic injuries and fatalities have increased alongside increased use of motorised transport in China, and accident injury risk is aggravated by inadequate emergency response systems and trauma care. National air quality standards and emission control technologies are having a positive effect on air quality, but persistent air pollution is increasingly attributable to a growing and outdated vehicle fleet and to famously congested roads. Urban design favours motorised transport, and physical activity and its associated health benefits are hindered by poor urban infrastructure. Transport emissions of greenhouse gases contribute substantially to regional and global climate change, which compound public health risks from multiple factors. Despite these complex challenges, technological advances and innovations in planning and policy stand to make China a leader in sustainable, healthy transportation.


Single-wavelength phototheranostics for colon cancer via the thiolytic reaction.
It's a huge challenge to develop effective nanosystems that combine the capabilities of diagnoses and therapies together for colon cancer in the clinic. Herein, we constructed a far-red absorbing phototheranostic nanosystem (FR-H2S) based on the thiolytic reaction of a dinitrophenyl modified phototheranostic prodrug and over-expressed H2S in colon cancer sites for precise imaging-guided phototherapy. FR-H2S with a BODIPY core not only could work as an imaging probe for diagnosis but also act as a phototherapeutic agent for cancer treatment under a single FR laser source (650 nm). FR-H2S exhibited a gradually enhanced fluorescence emission for precise diagnosis of H2S-rich colon tumor sites. After entering tumor cells, FR-H2S could generate abundant 1O2 and heat for phototherapies timely by using the same laser source (650 nm). We believe that this precise imaging-guided phototheranostic nanosystem could provide a promising approach to colon cancer with minimal damage.


[Reliability and construct validity of an instrument to asses the Self-perception of Family Health Status].
Family health determines and it is determined by family´s capacity to function effectively as a biosocial unit in a given culture and society. The main of study has been to test reliability and construct validity of an instrument to asses the Self-perception of Family Health Status. We validated its content by an on-line Dephi panel with experts. We surveyed 258 families in them homes or in primary health centres from Seville, Spain. We administered the instrument that has five Likert scales: Family climate, Family integrity, Family functioning, and Family resistance. We tested reliability by Cronbach Alpha and construct validity by exploratory factor analysis. The five scales obtained values α between 0.73 for the Family Climate and 0.89 for Family Integrity. They showed evidence of one-dimensional interpretation after factor analysis, a) all items got weights r>0.30 in first factor before rotations, b) the first factor explained a significant proportion of variance before rotations, and c) the total variance explained by the main factors extracted was greater than 50%. The scales showed their reliability and validity. They could be employed to assess the self-perception of family health status.


Tracking the Fate of Endogenous Retrovirus Segregation in Wild and Domestic Cats.
Endogenous retroviruses (ERVs) of domestic cats (ERV-DCs) are one of the youngest feline ERV groups in domestic cats (Felis silvestris catus); some members are replication competent (ERV-DC10, ERV-DC18, and ERV-DC14), produce the antiretroviral soluble factor Refrex-1 (ERV-DC7 and ERV-DC16), or can generate recombinant feline leukemia virus (FeLV). Here, we investigated ERV-DC in European wildcats (Felis silvestris silvestris) and detected four loci: ERV-DC6, ERV-DC7, ERV-DC14, and ERV-DC16. ERV-DC14 was detected at a high frequency in European wildcats; however, it was replication defective due to a single G → A nucleotide substitution, resulting in an E148K substitution in the ERV-DC14 envelope (Env). This mutation results in a cleavage-defective Env that is not incorporated into viral particles. Introduction of the same mutation into feline and murine infectious gammaretroviruses resulted in a similar Env dysfunction. Interestingly, the same mutation was found in an FeLV isolate from naturally occurring thymic lymphoma and a mouse ERV, suggesting a common mechanism of virus inactivation. Refrex-1 was present in European wildcats; however, ERV-DC16, but not ERV-DC7, was unfixed in European wildcats. Thus, Refrex-1 has had an antiviral role throughout the evolution of the genus Felis, predating cat exposure to feline retroviruses. ERV-DC sequence diversity was present across wild and domestic cats but was locus dependent. In conclusion, ERVs have evolved species-specific phenotypes through the interplay between ERVs and their hosts. The mechanism of viral inactivation may be similar irrespective of the evolutionary history of retroviruses. The tracking of ancestral retroviruses can shed light on their roles in pathogenesis and host-virus evolution.IMPORTANCE Domestic cats (Felis silvestris catus) were domesticated from wildcats approximately 9,000 years ago via close interaction between humans and cats. During cat evolution, various exogenous retroviruses infected different cat lineages and generated numerous ERVs in the host genome, some of which remain replication competent. Here, we detected several ERV-DC loci in Felis silvestris silvestris Notably, a species-specific single nucleotide polymorphism in the ERV-DC14 env gene, which results in a replication-defective product, is highly prevalent in European wildcats, unlike the replication-competent ERV-DC14 that is commonly present in domestic cats. The presence of the same lethal mutation in the env genes of both FeLV and murine ERV provides a common mechanism shared by endogenous and exogenous retroviruses by which ERVs can be inactivated after endogenization. The antiviral role of Refrex-1 predates cat exposure to feline retroviruses. The existence of two ERV-DC14 phenotypes provides a unique model for understanding both ERV fate and cat domestication.


Mode-of-action of the natural herbicide radulanin A as an inhibitor of photosystem II.
Radulanin A is a natural 2,5-dihydrobenzoxepin synthesized by several liverworts of the Radula genus. Breakthroughs in the total synthesis of radulanin A paved the way for the discovery of its phytotoxic activity. Nevertheless, its mode-of-action (MoA) has remained unknown so far and thus was investigated, in Arabidopsis thaliana. Radulanin A phytotoxicity was associated with cell death and partially depended on light exposure. Photosynthesis measurements based on chlorophyll-a fluorescence evidenced that radulanin A and a Radula chromene inhibited photosynthetic electron transport with IC50 of 95 and 100 μm, respectively. We established a strong correlation between inhibition of photosynthesis and phytotoxicity for a range of radulanin A analogs. Based on these data, we also determined that radulanin A phytotoxicity was abolished when the hydroxyl group was modified, and was modulated by the presence of the heterocycle and its aliphatic chain. Thermoluminescence studies highlighted that radulanin A targeted the QB site of the Photosystem II (PSII) with a similar MoA as 3-(3,4-dichloropheny)-1,1-dimethylurea (DCMU). We establish that radulanin A targets PSII, expanding QB sites inhibitors to bibenzyl compounds. The identification of an easy-to-synthesize analog of radulanin A with similar MoA and efficiency might be useful for future herbicide development. © 2023 Society of Chemical Industry.