Prospective care of elderly patients in family practice. Is screening effective?
To evaluate cost and benefits of screening for and treating health and lifestyle risks among community-dwelling elderly. Randomized controlled trial. Primary care. An opportunistic and prompted sample of 619 rostered elderly patients presenting for treatment who screened positive. One third (209) of experimental subjects had screening questionnaires placed in their charts with concerns highlighted for referrals. Two control groups received usual care. Yearly assessments of health service use and multidimensional functional capacity. Overall, screening and treatment of functionally active, elderly, middle-class people had no significant beneficial effect. Almost half of the experimental sample was ineligible because of treatment noncompliance. Generally ineligible subjects were older and more severely impaired. Subjects 75 years and older with risk factors showed improvement in daily living activities, and those living alone were found to have improved mental health and social functions (11% and 22%, respectively). Screening and treatment was ineffective in improving total functional capacity of all seniors 65 years and older. Elderly people 75 years and older, however, who were living alone or lonely did benefit from screening and treatment showing an improvement in daily activities, mental health scores, and social functions. This finding has implications for selective preventive health care spending for the elderly. A 2-year follow-up period could be too brief to detect long-term effects of early intervention with younger, middle-class seniors, especially those who are already functionally active.


Validity of death certificates in the identification of cases of amyotrophic lateral sclerosis (ALS) in the Limousin region, France. A population-based study.
Objectives: Mortality data have been used as a proxy for amyotrophic lateral sclerosis (ALS) incidence. However, the accuracy of death certificates (DCs) needs to be validated. This study aims to assess the accuracy of DCs in the identification of ALS cases. Methods: This is a retrospective population-based validation study. DC information, provided by the "Centre d'épidémiologie sur les causes médicales de décès", including ICD10 codes for specific cause of death for patients recorded in the French register of ALS cases in the Limousin region (FRALim) and deceased between 2000 and 2011, was assessed. The FRALim register was used as gold standard. Results: In the study period, DCs were available for 197 patients diagnosed with ALS, of whom 185 (93.9%) were correctly identified with an ICD10 code (G12.2) corresponding to ALS. The overall sensitivity was 93.9% (95% CI 89.6-96.8) and the positive predictive value (PPV) was 64.9 (59.1-70.4), with higher values in the period 2004-2011 (75.0-78.9). Stratification for sex, age, and year at death did not show difference in accuracy, except a lower PPV during the first years of observation. Conclusions: DCs identifying subjects with a diagnosis of ALS in the Limousin region, France showed an overall good sensitivity and moderate PPV. The absence of ALS diagnosis as the main and underlying cause of death on DCs highlights the need to use DC in combination with other administrative data to create algorithms with higher accuracy performances.


Adenovirus vector library: an approach to the discovery of gene and protein function.
A method was developed to generate a complex cDNA expression library within an adenovirus type 5 (Ad5)-based vector backbone, termed AdLibrary. Construction of the AdLibrary entailed the conversion of an Ad5 genome-containing cosmid to infectious virus particles. The Ad5 genome was modified by replacing the E1A and E1B genes with a Rous sarcoma virus-driven expression cassette. Conversion was accomplished by liberating the viral genome by restriction enzyme digestion and transfection in HEK 293 cells, which support the growth of E1A/E1B-deficient virus. A test AdLibrary demonstrated the possibility of converting and identifying a marker gene present at a frequency of 1/10(5) in the cosmid library. To demonstrate the utility of this technology, an AdLibrary was used to isolate a viral gene by its biological function. Virus growth was selected for with an AdLibrary on A549 cells, which do not complement for E1A/E1B function. The AdLibrary was generated with cDNAs derived from HeLa cells productively infected with Ad5. A cDNA corresponding to Ad5 E1A 13S was selected and isolated from the AdLibrary using this strategy. Since multiple genes are assayed simultaneously, this technology should expedite the discovery of genes affecting defined biological activities. This AdLibrary approach provides an opportunity to exploit the efficient gene delivery capabilities of adenovirus vectors for the rapid discovery of gene and protein function.


Prevention of cytosolic IAPs degradation: a potential pharmacological target in Huntington's Disease.
Huntington's Disease (HD) is a neurodegenerative disorder caused by an abnormally expanded polyglutamine trait in the amino-terminal region of huntingtin. Pathogenic mechanisms involve a gained toxicity of mutant huntingtin and a potentially reduced neuroprotective function of the wild-type allele. Among the molecular abnormalities reported, HD cells are characterized by the presence of aggregates, transcriptional dysregulation, altered mitochondrial membrane potential and aberrant Ca++ handling. In addition, upon exposure to toxic stimuli, increased mitochondrial release of cytochrome C and activation of caspase-9 and caspase-3 are found in HD cells and tissue. Here we report that HTRA2 and Smac/DIABLO, two additional mitochondrial pro-apoptotic factors, are aberrantly released from brain-derived cells expressing mutant huntingtin. This event causes a reduction in levels of the cytosolic IAP1 (Inhibitor of Apoptosis Protein-1) and XIAP (X-linked inhibitor apoptosis) antiapoptotic IAP family members. Reduced IAP levels are also found in post-mortem HD brain tissue. Treatment with ucf101, a serine protease HTRA2 specific inhibitor, counteracts IAPs degradation in HD cells and increases their survival. These results point to the IAPs as potential pharmacological targets in Huntington's Disease.


Both localized and systemic bacterial infections are predicted by injection drug use: A prospective follow-up study in Swedish criminal justice clients.
Both skin and soft tissue infections (SSTI) and systemic bacterial infections are common in people who inject drugs (PWID), but data on incidence and risk factors are lacking. We compared registered diagnoses for such infections in Swedish criminal justice clients with regard to injecting drug use. Baseline interview data from the Swedish Prison and Probation Service on drug use in PWID and non-PWID with problematic alcohol use were linked to follow-up data from national Swedish registers on hospital diagnoses and/or death. Associations between drug use and later diagnosis of SSTI and systemic bacterial infection (septicemia or bacterial infection of the heart, bone/joints or central nervous system) were analyzed by Cox regression. Incidence rates of SSTI was 28.3 per 1,000 person-years for PWID (n = 2,444) and 10.0 for non-PWID with problematic alcohol use (n = 735). Incidence rates of systemic bacterial infection was 9.1 per 1,000 person-years for PWID and 2.7 per 1,000 person-years for non-PWID. Injection drug use was associated with a significantly increased risk of bacterial infections, for main drugs heroin (SSTI: Hazard ratio [HR] 2.45; systemic infection: HR 2.75), amphetamine (SSTI: HR 1.60; systemic infection: HR 2.19), and polysubstance use (SSTI: HR 1.92; systemic infection: HR 2.01). In relation to injection use of amphetamine and polysubstance use, PWID mainly using heroin had higher risk of SSTI. Injection drug use predicted both SSTI and systemic bacterial infection, with a particularly high risk of SSTI in PWID mainly using heroin. The results imply the need for increased attention to bacterial infections among PWID, in terms of clinical management, prevention and research.


The application of minisatellite variant repeat mapping by PCR (MVR-PCR) in a paternity case showing false exclusion due to STR mutation.
A boy and a girl with their mother brought a paternity suit against an alleged but deceased father. We tested six conventional genetic markers, the AmpliType PM+ DQA1 and twelve STR loci the children and mother together with the alleged paternal grandparents. We also DNA typed the bloodstain found later in the alleged father's medical record. Only the result at D3S1358 in a nineplex STR system excluded the alleged father from parentage of the boy, whereas all markers were inclusive for the girl. Accordingly, we performed sequence analysis at D3S1358 to confirm the presence of a paternal exclusion or mutation. The sequence analysis indicated that the boy's allele 17 could have originated from either of the alleged father's allele 16 or 18 by a single-step mutation associated with slippage mutation in STR loci. We carried out minisatellite variant repeat mapping by PCR (MVR-PCR) at loci D1S8 (MS32) and D7S21 (MS31A) and mapped allele haplotypes of all individuals except the deceased alleged father. The MVR-PCR analysis showed that the boy has no inconsistency with the relationship between the alleged grandparents, and was very effective at increasing the paternity index (PI) value. We conclude that there is biological relationship between not only the girl but also the boy and the alleged father.


Exercise hemodynamics enhance diagnosis of early heart failure with preserved ejection fraction.
When advanced, heart failure with preserved ejection fraction (HFpEF) is readily apparent. However, diagnosis of earlier disease may be challenging because exertional dyspnea is not specific for heart failure, and biomarkers and hemodynamic indicators of volume overload may be absent at rest. Patients with exertional dyspnea and ejection fraction >50% were referred for hemodynamic catheterization. Those with no significant coronary disease, normal brain natriuretic peptide assay, and normal resting hemodynamics (mean pulmonary artery pressure <25 mm Hg and pulmonary capillary wedge pressure [PCWP] <15 mm Hg) (n=55) underwent exercise study. The exercise PCWP was used to classify patients as having HFpEF (PCWP ≥25 mm Hg) (n=32) or noncardiac dyspnea (PCWP <25 mm Hg) (n=23). At rest, patients with HFpEF had higher resting pulmonary artery pressure and PCWP, although all values fell within normal limits. Exercise-induced elevation in PCWP in HFpEF was confirmed by greater increases in left ventricular end-diastolic pressure and was associated with blunted increases in heart rate, systemic vasodilation, and cardiac output. Exercise-induced pulmonary hypertension was present in 88% of patients with HFpEF and was related principally to elevated PCWP, as pulmonary vascular resistances dropped similarly in both groups. Exercise PCWP and pulmonary artery systolic pressure were highly correlated. An exercise pulmonary artery systolic pressure ≥45 mm Hg identified HFpEF with 96% sensitivity and 95% specificity. Euvolemic patients with exertional dyspnea, normal brain natriuretic peptide, and normal cardiac filling pressures at rest may have markedly abnormal hemodynamic responses during exercise, suggesting that chronic symptoms are related to heart failure. Earlier and more accurate diagnosis using exercise hemodynamics may allow better targeting of interventions to treat and prevent HFpEF progression.


Endotoxic shock elicits greater endorphin secretion than hemorrhage.
Opiopeptides may contribute to the pathophysiology of both endotoxic and hemorrhagic shock. To determine if endorphin secretion is similar in both types of shock, we divided 25 sheep into three groups: a saline control group (n = 10), an endotoxin-treated group (n = 9), and a hemorrhage group (n = 6). Each sheep had baseline determinations of mean arterial pressure (MAP) and plasma levels of beta-endorphin-like immunoreactivity (iB-EP). Experimental animals either received endotoxin (450 ng/kg intravenous) or underwent withdrawal of blood volume sufficient to diminish MAP by approximately one-third of baseline values. MAP and iB-EP levels were determined every 15 minutes thereafter for 5 hours. Individual data were averaged within each group and then compared between groups using analysis of variance. Both the endotoxin- and hemorrhage-treated groups showed a significant fall in MAP, which was significantly lower in the hemorrhage group than the endotoxin group. Endotoxin-treated animals displayed a mean peak iB-EP level 1,550% above baseline as compared to a mean peak iB-EP level of only 201% above baseline in the hemorrhage-treated group, despite a significantly greater degree of hypotension in the latter group; this difference in peak iB-EP response was significant. Mean peak iB-EP levels coincided with mean trough MAP values in the endotoxin-treated group while the mean peak iB-EP lagged the onset of mean trough MAP in the hemorrhage group. These results demonstrate that iB-EP secretory patterns differ in endotoxic versus hemorrhagic shock and suggest that distinct mechanisms of opiopeptide secretion accompany the two shock states.


A Rapid and Automated Sample-to-Result Candida auris Real-Time PCR Assay for High-Throughput Testing of Surveillance Samples with the BD Max Open System.
The multidrug-resistant yeast pathogen Candida auris continues to cause outbreaks and clusters of clinical cases worldwide. Previously, we developed a real-time PCR assay for the detection of C. auris from surveillance samples (L. Leach, Y. Zhu, and S. Chaturvedi, J Clin Microbiol 56:e01223-17, 2018, https://doi.org/10.1128/JCM.01223-17). The assay played a crucial role in the ongoing investigations of the C. auris outbreak in New York City. To ease the implementation of the assay in other laboratories, we developed an automated sample-to-result real-time C. auris PCR assay using the BD Max open system. We optimized sample extraction at three different temperatures and four incubation periods. Sensitivity was determined using eight pools of patient samples, and specificity was calculated using four clades of C. auris and closely and distantly related yeasts. Three independent extractions and testing of two patient sample pools in quadruplicate yielded assay precision. BD Max optimum assay conditions were as follows: DNA extraction at 75°C for 20 min and the use of PerfeCTa multiplex quantitative PCR (qPCR) ToughMix. The limit of detection (LOD) of the assay was one C. auris CFU/PCR. We detected all four clades of C. auris without cross-reactivity to other yeasts. Of the 110 patient surveillance samples tested, 50 were positive for C. auris using the BD Max system with 96% clinical sensitivity and 94% accuracy compared to the results of the manual assay. The BD Max assay allows high-throughput C. auris screening of 180 surveillance samples in a 12-h workday.


Retention through redemption.
Corporate America and the U.S. Navy share one big problem: employee retention. Today's knowledge workers hop from start-up to start-up. And 40% of the navy's new recruits leave the service before their four-year tours of duty are up. D. Michael Abrashoff came face to face with the navy's retention problem when he took command of the USS Benfold. Before he became captain, sailors couldn't get away from the ship fast enough. Today the vessel is the pride of the Pacific fleet, and sailors from other ships are clamoring to join its crew. In this firsthand account, Abrashoff explains how he got the ship and its crew back on course by breaking bad habits--personal and professional ones--and jettisoning old attitudes. During his 21 months aboard the Benfold, Abrashoff came to realize that in today's technology-intensive U.S. Navy, the traditional command-and-control style wouldn't work. And it hadn't--the Benfold's 310 sailors had cheered derisively when Abrashoff's predecessor had left the ship. So he defied 225 years of navy tradition in his quest to engage the sailors in their work, increase their performance, and keep them around for their entire tours of duty. He retained his crew by redeeming them--showing them how to be not just better sailors but better people, too. That meant breaking them down when they were at their worst and then building them up to reach their best. It also meant personal redemption for Abrashoff; he resolved to really listen to what his sailors were saying. The result? Cost-saving ideas for the entire navy and surging confidence and commitment among crew members.


Role of mucoadhesive polymers in retention of toothpaste in the oral cavity.
Retention of active ingredients of toothpastes in the mouth following brushing determines the efficiency of these oral care formulations. In this study, new in vitro methodologies for the observation and measurement of toothpaste retention in the oral cavity were developed and used to evaluate the efficiency of formulations containing different mucoadhesive hydrophilic polymers. The findings suggest that using Carbopol ETD 2020 and Carbopol Ultrez 10 as binders in toothpaste prolongs the retention time of these formulations in the oral cavity. The in vitro methodologies tested, coupled with texture analysis, were able to accurately characterise the behaviour of the toothpaste and produce detailed images showing how it is retained in the oral cavity. This study has not only produced a new method for studying the behaviour of toothpaste and other formulations in the oral cavity but is also the first to investigate how different types of mucoadhesive binders can be used to improve toothpaste retention.


Arsenic downregulates tight junction claudin proteins through p38 and NF-κB in intestinal epithelial cell line, HT-29.
Arsenic is a naturally occurring metalloid that often is found in foods and drinking water. Human exposure to arsenic is associated with the development of gastrointestinal problems such as fluid loss, diarrhea and gastritis. Arsenic is also known to induce toxic responses including oxidative stress in cells of the gastrointestinal track. Tight junctions (TJs) regulate paracellular permeability and play a barrier role by inhibiting the movement of water, solutes and microorganisms in the paracellular space. Since oxidative stress and TJ damage are known to be associated, we examined whether arsenic produces TJ damage such as downregulation of claudins in the human colorectal cell line, HT-29. To confirm the importance of oxidative stress in arsenic-induced TJ damage, effects of the antioxidant compound (e.g., N-acetylcysteine (NAC)) were also determined in cells. HT-29 cells were treated with arsenic trioxide (40μM, 12h) to observe the modified expression of TJ proteins. Arsenic decreased expression of TJ proteins (i.e., claudin-1 and claudin-5) and transepithelial electrical resistance (TEER) whereas pretreatment of NAC (5-10mM, 1h) attenuated the observed claudins downregulation and TEER. Arsenic treatment produced cellular oxidative stress via superoxide generation and lowering glutathione (GSH) levels, while NAC restored cellular GSH levels and decreased oxidative stress. Arsenic increased phosphorylation of p38 and nuclear translocation of nuclear factor-kappa B (NF-κB) p65, while NAC attenuated these intracellular events. Results demonstrated that arsenic can damage intestinal epithelial cells by proinflammatory process (oxidative stress, p38 and NF-κB) which resulted in the downregulation of claudins and NAC can protect intestinal TJs from arsenic toxicity.


Clinical potential of antigen-specific therapies in type 1 diabetes.
In type 1 diabetes (T1D), pancreatic beta-cells are attacked and destroyed by the immune system, which leads to a loss of endogenous insulin secretion. The desirable outcome of therapeutic intervention in autoimmune diseases is the restoration of immune tolerance to prevent organ damage. Past trials with immune suppressive drugs highlight the fact that T1D is in principle a curable condition. However, the barrier in T1D therapy in terms of drug safety is set particularly high because of the predominantly young population and the good prognosis associated with modern exogenous insulin therapy. Thus, there is a general consensus that chronic immune suppression is associated with unacceptable long-term safety risks. On the other hand, immune-modulatory biologicals have recently failed to confer significant protection in phase 3 clinical trials. However, the concept of antigen-specific tolerization may offer a unique strategy to safely induce long-term protection against T1D. In this review, we analyze the potential reasons for the failure of the different tolerization therapies, and describe how the concept of antigen-specific toleraization may overcome the obstacles associated with clinical therapy in T1D.


Effect of co-transporter blockers on non-synaptic epileptiform activity-computational simulation.
The important role of cation-chloride co-transporters in epilepsy is being supported by an increasing number of investigations. However, enormous complexity is involved since the action of these co-transporters has effects on the ionic homeostasis influencing directly the neuronal excitability and the tissue propensity to sustain seizure. To unravel the complex mechanisms involving the co-transporters action during seizure, this paper shows simulations of non-synaptic epileptiform activity and the effect of the blockage of the two different types of cation-chloride co-transporters present in the brain: Na, K and 2Cl co-transporter (NKCC) and K and Cl co-transporter (KCC). The simulations were performed with an electrochemical model representing the non-synaptic structure of the granule cell layer of the dentate gyrus (DG) of the rat hippocampus. The simulations suggest: (i) the potassium clearance is based on the systemic interplay between the Na/K pump and the NKCC co-transporters; (ii) the simultaneous blockage of the NKCC of the neurons and KCC of glial cells acts efficiently suppressing the epileptiform activities; and (iii) the simulations show that depending on the combined blockage of the co-transporters, the epileptiform activities may be suppressed or enhanced.


Hospital volume is associated with postoperative mortality after radical cystectomy for treatment of bladder cancer.
To contribute to the debate regarding the minimum volume of radical cystectomies (RCs) that a hospital should perform by evaluating the association between hospital volume (HV) and postoperative mortality. Patients who underwent RC for bladder cancer between 1 January 2008 and 31 December 2018 were retrospectively identified from the Netherlands Cancer Registry. To create a calendar-year independent measure, the HV of RCs was calculated per patient by counting the RCs performed in the same hospital in the 12 months preceding surgery. The relationship of HV with 30- and 90-day mortality was assessed by logistic regression with a non-linear spline function for HV as a continuous variable, which was adjusted for age, tumour, node and metastasis (TNM) stage, and neoadjuvant treatment. The median (interquartile range; range) HV among the 9287 RC-treated patients was 19 (12-27; 1-75). Of all the included patients, 208 (2.2%) and 518 (5.6%) died within 30 and 90 days after RC, respectively. After adjustment for age, TNM stage and neoadjuvant therapy, postoperative mortality slightly increased between an HV of 0 and an HV of 25 RCs and steadily decreased from an HV of 30 onwards. The lowest risks of postoperative mortality were observed for the highest volumes. This paper, based on high-quality data from a large nationwide population-based cohort, suggests that increasing the RC volume criteria beyond 30 RCs annually could further decrease postoperative mortality. Based on these results, the volume criterion of 20 RCs annually, as recently recommended by the European Association of Urology Guideline Panel, might therefore be reconsidered.


[Minimally invasive surgical methods in treatment of nodular goiter].
The article presents results of topographo-anatomical investigation of the asymmetric mini-access used in surgical treatment of the nodular goiter. It was established that the mini-access is rational, adequate and does not require special surgical instruments. The performance of hemithyroidectomy from the asymmetric mini-access used in 99 patients with nodular goiter had excellent cosmetic effect, early rehabilitation and low number of complications. Long-term results of treatment by the method of ethanol destruction were investigated in 50 patients. At the end of the observation the diameter of the nodes was found to increase as compared with the minimal indices.


Security of attachment and infant-sibling relationships: a laboratory study.
The relation between infant-sibling affective involvement and the attachment security of each child to the mother was examined in the present laboratory investigation. In mothers' presence, securely attached infants were less likely to protest and aggress against mothers and older siblings when mothers played only with the older child. In mothers' absence, more secure older siblings were more likely to respond to infant distress with caregiving than were less secure older siblings. Although infant attachment behavior to older siblings was rare, it occurred only when the older sibling was more secure. Sibling dyads with a secure infant and a more secure older child appeared to be most likely to develop nonantagonistic relationships, whereas sibling dyads with an insecure infant and a less secure older child appeared least likely to do so. These findings were discussed in terms of the putative role of attachment security in shaping sibling bonds.


Puromycin-purified rat brain microvascular endothelial cell cultures exhibit improved barrier properties in response to glucocorticoid induction.
In vitro blood-brain barrier (BBB) models using primary rat brain microvessel endothelial cells (BMEC) are often hampered by a lack of culture purity and poor barrier properties. To address these problems, the translation inhibitor puromycin was used to purify rat BMEC cultures. BMEC purities of 99.8% were routinely attained using puromycin treatment, and this technique proved to be far superior to other purification methods of similar difficulty. In contrast to cultures without puromycin treatment, purity of puromycin-treated cultures was unaffected by initial seeding density. Next, rat BMEC monolayer transendothelial electrical resistance (TEER) was increased by glucocorticoid treatment with either corticosterone (CORT) or hydrocortisone (HC), and a corresponding decrease in monolayer permeability to small molecules was observed. Importantly, cultures treated with both puromycin and glucocorticoid attained significantly higher TEER values (CORT 168 +/- 13 Omega x cm2; HC 218 +/- 66 Omega x cm2) than those treated by the glucocorticoid alone (CORT 57 +/- 5 Omega x cm2; HC 70 +/- 2 Omega x cm2). Glucocorticoid induction resulted in BMEC morphological changes that accompanied the increases in TEER, and BMEC tight junctions exhibited improved integrity as visualized by the localization of tight junction proteins zonula occluden-1, occludin and claudin-5. The combined use of puromycin and glucocorticoid therefore provides an in vitro system that is well suited for molecular level BBB investigations.


Targeted histone acetylation and altered nuclease accessibility over short regions of the pea plastocyanin gene.
The chromatin structure of the pea plastocyanin gene (PetE) was examined at three different transcriptional states by investigating the acetylation states of histones H3 and H4 and the nuclease accessibility of the gene in pea roots, etiolated shoots, and green shoots. The acetylation states of histones associated with different regions of PetE were analyzed by chromatin immunoprecipitation with antibodies specific for acetylated or nonacetylated histone H3 or H4 tails, followed by polymerase chain reaction quantification. Comparison of pea tissues indicated that histone hyperacetylation was associated with increased PetE transcription in green shoots. Moreover, hyperacetylation of both histones H3 and H4 was targeted to the enhancer/promoter region in green shoots, suggesting that only specific nucleosomes along the gene were modified. Time-course digestions of nuclei with micrococcal nuclease and DNaseI indicated that the enhancer/promoter region was more resistant to digestion in the inactive gene in pea roots than was the same region in the active gene in shoots, whereas the transcribed region of PetE was digested similarly among the tissues. This finding indicates that transcription is accompanied by changes in the nuclease accessibility of the enhancer/promoter region only. Moreover, these results indicate that the changes in nuclease accessibility are organ specific, whereas histone hyperacetylation is light dependent, and they suggest that changes in nuclease accessibility precede histone hyperacetylation during PetE activation.


Fibronectin conformation switch induced by coadsorption with human serum albumin.
The dynamic adsorption of human serum albumin (HSA) and plasma fibronectin (Fn) onto hydrophobic poly(hydroxymethylsiloxane) (PHMS) and the structures of adsorbed protein layers from single and binary protein solutions were studied. Spectroscopic ellipsometry (SE) and quartz crystal microbalance with dissipation monitoring (QCM-D) together with atomic force microscopy (AFM) were used to measure the effective mass, thickness, viscoelastic properties, and morphology of the adsorbed protein films. Adsorbed HSA formed a rigid, tightly bound monolayer of deformed protein, and Fn adsorption yielded a thick, very viscoelastic layer that was firmly bound to the substrate. The mixed protein layers obtained from the coadsorption of binary equimolecular HSA-Fn solutions were found to be almost exclusively dominated by Fn molecules. Further sequential adsorption experiments showed little evidence of HSA adsorbed onto the predeposited Fn layer (denoted as Fn ≫ HSA), and Fn was not adsorbed onto predeposited HSA (HSA ≫ Fn). The conformational arrangement of the adsorbed Fn was analyzed in terms of the relative availability of two Fn domains. In particular, (4)F(1)·(5)F(1) binding domains in the Hep I fragment, close to the amino terminal of Fn, were targeted using a polyclonal antifibronectin antibody (anti-Fn), and the RGD sequence in the 10th segment, in the central region of the molecule, was tested by cell culture experiments. The results suggested that coadsorption with HSA induced the Fn switch from an open conformation, with the amino terminal subunit oriented toward the solution, to a close conformation, with the Fn central region oriented toward the solution.


Effect of magnetic field on the hydrogen evolution activity using non-magnetic Weyl semimetal catalysts.
An external switch to control the kinetics of the reaction by manipulating the participating electrons could be interesting as it can alter the rate of the reaction without affecting the reaction pathway. The magnetic field, like a switch, is non-invasive, tunable, and clean; it can also alter the electrons in a material. We study the effect of an applied magnetic field on the hydrogen evolution activity of the NbP family of Weyl semimetals because of their extremely high mobility and large magnetoresistance at room temperature and good hydrogen evolution properties. We find that by applying a magnetic field of ∼3500 G, the hydrogen evolution activity of NbP increases by up to 95%. The other members of this Weyl semimetal family (viz. TaP, NbAs, and TaAs) also exhibit increased hydrogen evolution activity. Thus, our observations suggest an interplay of electronic property, magnetic field, and catalytic activity in this class of compounds, providing evidence of manipulating the catalytic performance of topological materials through the application of a magnetic field.


Effect of peer led programme for asthma education in adolescents: cluster randomised controlled trial.
To determine the effect of a peer led programme for asthma education on quality of life and related morbidity in adolescents with asthma. Cluster randomised controlled trial. Six high schools in rural Australia. 272 students with recent wheeze, recruited from a cohort of 1515 students from two school years (mean age 12.5 and 15.5 years); 251 (92.3%) completed the study. A structured education programme for peers comprising three steps (the "Triple A Program"). Quality of life, school absenteeism, asthma attacks, and lung function. When adjusted for year and sex, mean total quality of life scores showed significant improvement in the intervention than control group. Clinically important improvement in quality of life (>0.5 units) occurred in 25% of students with asthma in the intervention group compared with 12% in the control group (P=0.01). The number needed to treat was 8 (95% confidence interval 4.5 to 35.7). The effect of the intervention was greatest in students in year 10 and in females. Significant improvements occurred in the activities domain (41% v 28%) and in the emotions domain (39% v 19%) in males in the intervention group. School absenteeism significantly decreased in the intervention group only. Asthma attacks at school increased in the control group only. The triple A programme leads to a clinically relevant improvement in quality of life and related morbidity in students with asthma. Wider dissemination of this programme in schools could play an important part in reducing the burden of asthma in adolescents.


Nesting behavior of the Picazuro pigeon, Columba Picazuro (Columbidae, Aves).
The Picazuro Pigeon nests in all months of the year in southeastern Brazil. Nest material is plucked from trees or ground and carried to build a frail and transparent nest of sticks where one egg is laid. Female and male alternate in incubation and brooding and do not soil the nest with feces.


Gluten and skin disease beyond dermatitis herpetiformis: a review.
Gluten, a protein found in wheat, rye, and barley, is known to cause an immune reaction in patients with celiac disease (CD) resulting in small bowel villous atrophy and impaired nutrient absorption and cutaneous manifestations in patients with dermatitis herpetiformis (DH). It is common that patients associate skin conditions with their diet, and the advantages of a gluten-free diet (GFD) are brought up frequently. Indeed, there is evidence that certain dermatologic conditions can respond to a GFD, especially for those with concomitant CD and DH. In the last decade, new data have become available on the significance of gluten in skin disease. Herein, we review the role of gluten and a GFD on various cutaneous diseases beyond DH.


Association of health behaviors, dietary habits, and oral health with weight gain after 20 years of age in community-dwelling Japanese individuals aged 40 years and older: a cross-sectional study.
This cross-sectional study investigated the factors associated with weight gain ≥ 10 kg after 20 years of age in the general Japanese population, with a focus on the number of teeth. We included individuals aged ≥ 40 years from Yamagata prefecture, Japan from 2017-2021. A postal survey was conducted using a self-administered questionnaire; 5,940 participants were included in the final analysis. The questionnaire included items on lifestyle factors, medical history, physical and mental conditions, oral health, and dietary intake. A multivariate logistic regression analysis was used to identify independent associations between weight gain ≥ 10 kg after 20 years of age and various parameters; adjusted odds ratios (ORs) and 95% confidence intervals (CIs) were estimated. Less than 20 teeth, male sex, drinking habit frequency, eating very fast or fast, and a higher frequency of eating-away-from-home were significant factors associated with weight gain ≥ 10 kg after 20 years of age; individuals with < 20 versus > 20 teeth exhibited a 1.35-fold higher OR (95% CI 1.15-1.59; p < 0.01). Our results suggest that having < 20 teeth may affect weight gain ≥ 10 kg after 20 years of age. However, owing to the cross-sectional study design, causality could not be determined. Therefore, maintaining healthy lifestyle behaviors to avoid tooth loss may also affect weight gain ≥ 10 kg after 20 years of age. Having < 20 teeth has the potential to affect long-term weight gain after 20 years of age.


Risk factors and characteristics of post-transplant tuberculosis in an endemic area.
Tuberculosis (TB) is a major post-transplant concern in endemic areas. This report summarizes the clinical characteristics, risk factors, and effects of post-transplant TB on graft and patient survival at a single center in Korea. We retrospectively analyzed data from 2799 kidney recipients at Yonsei University Health System between April 1979 and August 2008 to determine the incidence, outcome, and risk factors affecting the development of TB infections and the effect of TB on graft and patient survival rates. TB developed in 144 (109 males and 35 females; mean age, 37 ± 12 years) out of 2799 (5.1%) recipients. Newly developed TB occurred in 116 recipients (81%) and recurrent TB occurred in 28 (19%) recipients with a pre-transplant history of a mycobacterial infection. The mean interval to TB diagnosis was 55.6 ± 47.9 months after transplantation without a peak interval incidence for 8 years after transplantation. Based on Cox regression analysis, a history of TB was the strongest risk factor (hazard ratio [HR] =11.618) affecting the development of TB. TB negatively affected graft and patient survival after kidney transplantation. Non-pleurisy extrapulmonary and miliary TB resulted in inferior treatment results and a poor prognosis in the early treatment period. A history of TB is the strongest predictor of post-transplant TB. Therefore, patients with a pre-transplant history of TB should be carefully monitored.


Altitude training and its influence on physical endurance in swimmers.
It is possible to plan an altitude training (AT) period in such a way that the enhanced physical endurance obtained as a result of adaptation to hypoxia will appear and can be used to improve performance in competition. Yet finding rationales for usage of AT in highly trained swimmers is problematic. In practice AT, in its various forms, is still controversial, and an objective review of research concentrating on the advantages and disadvantages of AT has been presented in several scientific publications, including in no small part the observations of swimmers. The aim of this article is to review the various methods and present both the advantageous and unfavourable physiological changes that occur in athletes as a result of AT. Moreover, AT results in the sport of swimming have been collected. They include an approach towards primary models of altitude/hypoxic training: live high + train high, live high + train low, live low + train high, as well as subsequent methods: Intermittent Hypoxic Exposure (IHE) and Intermittent Hypoxic Training (IHT). Apnoea training, which is descended from freediving, is also mentioned, and which can be used with, or as a substitute for, the well-known IHE or IHT methods. In conclusion, swimmers who train using hypoxia may be among the best-trained athletes, and that even a slight improvement in physical endurance might result in the shortening of a swimming time in a given competition, and the achievement of a personal best, which is hard to obtain by normal training methods, when the personal results of the swimmer have reached a plateau.


Seroprevalence of hepatitis B virus infection and associated factors among healthcare workers in northern Tanzania.
Hepatitis B virus infection is a global health problem with the highest prevalence in East Asia and Sub-Saharan Africa. The majority of infected people, including healthcare workers are unaware of their status. This study is aimed to determining seroprevalence of hepatitis B virus infection and associated factors among healthcare workers in northern Tanzania. This cross-sectional study included 442 healthcare workers (HCWs) from a tertiary and teaching hospital in Tanzania before the nationwide hepatitis B vaccination campaign in 2004. Questionnaire- based interviews were used to obtain detailed histories of the following: demographic characteristics; occupation risks such splash and needle stick injuries or other invasive procedure such as intravenous, intramuscular or subcutaneous injections; history of blood transfusion and surgeries, as well as HCWs'knowledge of HBV. Serological markers of HBV were done using Laborex HBsAg rapid test. Serology was done at zero months and repeated after six months ( bioscienceinternational.co.ke/rapid-test-laborex.html HBsAg Piazzale-milano-2, Italy [Accessed on November 2017]). Chi-square (χ2) tests were used to compare proportion of HBV infection by different HCWs characteristics. Multivariable logistic regression was used to determine factors associated with HBV infection. A total of 450 surveys were sent out, with a 98.2% response rate. Among the 442 HCWs who answered the questionnaire, the prevalence of chronic hepatitis B virus infection was 5.7% (25/442). Only 50 (11.3%) of HCWs were aware of the HBV status. During the second HBsAg testing which was done after six months one participant sero-converted hence was excluded. Adjusted for other factors, history of blood transfusion significantly increased the odds of HBV infection (OR = 21.44, 95%CI 6.05, 76.01, p < 0.001) while HBV vaccine uptake was protective against HBV infection (OR = 0.06, 95%CI 0.02, 0.26, p < 0.001). The majority of HCWs with chronic HBV infection had poor to fare knowledge about HBV infection but this was not statistically significant when controlled for confounding. Prevalence of HBV among health care workers was 5.7% which is similar to national prevalence. Although the response rate to take part in the study was good but knowledge on HBV infection among HCWs was unsatisfactory. History of blood transfusion increased risks while vaccine uptake decreased the risk of HBV infection. This study recommends continues vaccinating HCWs together with continues medical education all over the country. We also recommend documentation of vaccination evidence should be asked before employment of HCWs in order to sensitize more uptakes of vaccinations. Although we didn't assess the use of personal protective equipment but we encourage HCWs to abide strictly on universal protections against nosocomial infections.


Weighing the gold in the gold standard: challenges in HIV prevention research.
Few HIV prevention interventions have been evaluated in randomized controlled trials (RCTs). We examined design, implementation, and contextual considerations that may limit detection of a positive or adverse effect in HIV prevention trials. A systematic review of late phase RCTs for prevention of sexual transmission of HIV that randomly allocated intervention and comparison groups; evaluated interventions to prevent sexual transmission in nonpregnant populations; and reported HIV incidence as the primary or secondary outcome. PubMed/MEDLINE, other electronic databases, and electronic conference proceedings of recent HIV/AIDS-related conferences were searched to identify published or unpublished trials meeting the inclusion criteria. Descriptive, methodological, and contextual factors were abstracted from each trial. The review included 37 HIV prevention RCTs reporting on 39 unique interventions. Only six RCTs, all evaluating biomedical interventions, demonstrated definitive effects on HIV incidence. Five of the six RCTs significantly reduced HIV infection: all three male circumcision trials, one trial of sexually transmitted infection treatment and care, and one vaccine trial. One microbicide trial of nonoxynol-9 gel produced adverse results. Lack of statistical power, poor adherence, and diluted versions of the intervention in comparison groups may have been important issues for the other trials that demonstrated 'flat' results. Almost 90% of HIV prevention trials had 'flat' results, which may be attributable to trial design and/or implementation. The HIV prevention community must not only examine evidence from significant RCTs, but must also examine flat trials and address design and implementation issues that limit detection of an effect.


Extracts from presumed "reduced harm" cigarettes induce equivalent or greater toxicity in antigen-presenting cells.
The tobacco industry has promoted certain cigarette products with claims that their use may be less harmful to the smoker as they purportedly deliver lower amounts of toxic chemicals compared to conventional cigarettes. This study was designed to compare the relative antigen presenting cellular toxicity of Eclipse, a presumed reduced exposure product (PREP) cigarette, when compared with the reference research 3R4F cigarettes (Kentucky University). Utilizing a murine macrophage cell line, murine bone marrow derived dendritic cells (DCs) and human monocyte-derived DCs incubated with extracts generated from Eclipse and Kentucky reference 3R4F cigarettes, we determined the relative toxic effects of the different cigarette smoke extracts on cellular viability, oxidative stress, T-helper-1 (Th-1) polarizing cytokine production and general gene expression. Eclipse and 3R4F cigarette smoke extracts induced equivalent oxidatively-mediated cellular heme oxygenase-1 (HO-1) protein levels in macrophages and DCs. Cellular viability determination demonstrated greater induction of cell death by apoptosis and necrosis by Eclipse extracts in DCs. The production of the key Th-1 polarizing cytokine interleukin-12 (IL-12) by activated DCs or macrophages was suppressed to an equivalent or greater extent by Eclipse extracts. Microarray studies performed on bone marrow derived murine DCs incubated with Eclispe or 3R4F cigarette extracts showed identical genotoxic profiles. These studies imply that presumed reduced harm Eclipse cigarettes induce equivalent or greater antigen presenting cell dysfunction relative to 3R4F cigarettes and illustrate the importance of independent validation and testing of similar products claimed to be associated with reduced toxicity relative to other cigarettes.


An R195K Mutation in the PA-X Protein Increases the Virulence and Transmission of Influenza A Virus in Mammalian Hosts.
In the 21st century, the emergence of H7N9 and H1N1/2009 influenza viruses, originating from animals and causing severe human infections, has prompted investigations into the genetic alterations required for cross-species transmission. We previously found that replacement of the human-origin PA gene segment in avian influenza virus (AIV) could overcome barriers to cross-species transmission. Recently, it was reported that the PA gene segment encodes both the PA protein and a second protein, PA-X. Here, we investigated the role of PA-X. We found that an H9N2 avian influenza reassortant virus bearing a human-origin H1N1/2009 PA gene was attenuated in mice after the loss of PA-X. Reverse genetics analyses of PA-X substitutions conserved in human influenza viruses indicated that R195K, K206R, and P210L substitutions conferred significantly increased replication and pathogenicity on H9N2 virus in mice and ferrets. PA-X R195K was present in all human H7N9 and H1N1/2009 viruses and predominated in human H5N6 viruses. Compared with PA-X 195R, H7N9 influenza viruses bearing PA-X 195K showed increased replication and transmission in ferrets. We further showed that PA-X 195K enhanced lung inflammatory responses, potentially due to decreased host shutoff function. A competitive transmission study in ferrets indicated that 195K provides a replicative advantage over 195R in H1N1/2009 viruses. In contrast, PA-X 195K did not influence the virulence of H9N2 AIV in chickens, suggesting that the effects of the substitution were mammal specific. Therefore, future surveillance efforts should scrutinize this region of PA-X because of its potential impact on cross-species transmission of influenza viruses.IMPORTANCE Four influenza pandemics in humans (the Spanish flu of 1918 [H1N1], the Asian flu of 1957 [H2N2], the Hong Kong flu of 1968 [H3N2], and the swine origin flu of 2009 [H1N1]) are all proposed to have been caused by avian or swine influenza viruses that acquired virulence factors through adaptive mutation or reassortment with circulating human viruses. Currently, influenza viruses circulating in animals are repeatedly transmitted to humans, posing a significant threat to public health. However, the molecular properties accounting for interspecies transmission of influenza viruses remain unclear. In the present study, we demonstrated that PA-X plays an important role in cross-species transmission of influenza viruses. At least three human-specific amino acid substitutions in PA-X dramatically enhanced the adaptation of animal influenza viruses in mammals. In particular, PA-X 195K might have contributed to cross-species transmission of H7N9, H5N6, and H1N1/2009 viruses from animal reservoirs to humans.


Separation between the digestive and the respiratory lumina during the human embryonic period: morphometric study along the tracheo-oesophageal septum.
An isolated tracheo-oesophageal fistula could be caused by close proximity of the epithelia of both organs (O'Rahilly & Müller, 1984; Kluth et al. 1987) at certain embryonic stages, the most frequent location being the tracheal bifurcation. Thus the relative position and degree of separation between the digestive and the respiratory tubes throughout their development may be relevant to the origin of this anomaly. The aim of this study was to analyse along the different segments of the tracheo-oesophageal septum (TES) where the closest relationship between both lumina occurred and what degree of separation was present at each segment. Computer imaging techniques were applied on cross sections of a graded series of normal human embryos (Carnegie stages (CS) 13-23). In addition, the differentiation of the primitive TES was also studied (from CS 12) by light microscopy. Between CS 13 and 16 both tubes tended to separate (phase of separation), principally at the proximal segments of the laryngopharyngeal and the tracheo-oesophageal portions of the TES. During this phase the separation between the trachea and oesophagus was wider than between the larynx and pharynx. From CS 17 to CS 23 the digestive and respiratory lumina reached their widest separation at different levels of the laryngopharyngeal portion. Below these levels they tended to come closer together, principally at the proximal segment of the tracheo-oesophageal portion, but also at the distal part of the laryngopharyngeal portion. During this phase of approximation they reached their closest relationship at the proximal (CS 17) and the distal (from CS 18) segments of the tracheo-oesophageal portion. When finally the distal segment of the trachea (which includes the bifurcation) comes closest to the oesophagus, the coats of both organs have already undergone an appreciable differentiation. According to these observations, the origin of the most frequent isolated tracheo-oesophageal fistula at the bifurcation region could not be explained from the normal development of the TES.


Bacteria associated with a tree-killing insect reduce concentrations of plant defense compounds.
Bark beetles encounter a diverse array of constitutive and rapidly induced terpenes when attempting to colonize living conifers. Concentrations of these compounds at entry sites can rapidly reach levels toxic to beetles, their brood, and fungal symbionts. Large numbers of beetles can overwhelm tree defenses via pheromone-mediated mass attacks, but the mechanisms are poorly understood. We show that bacteria associated with mountain pine beetles can metabolize monoterpenes and diterpene acids. The abilities of different symbionts to reduce concentrations of different terpenes appear complementary. Serratia reduced concentrations of all monoterpenes applied to media by 55-75 %, except for α-pinene. Beetle-associated Rahnella reduced (-)- and (+)-α-pinene by 40 % and 45 %, respectively. Serratia and Brevundimonas reduced diterpene abietic acid levels by 100 % at low concentrations. However, high concentrations exhausted this ability, suggesting that opposing rates of bacterial metabolism and plant induction of terpenes are critical. The two major fungal symbionts of mountain pine beetle, Grosmannia clavigera and Ophiostoma montium were highly susceptible to abietic acid. Grosmannia clavigera did not reduce total monoterpene concentrations in lodgepole pine turpentine. We propose the ability of bark beetles to exert landscape-scale impacts may arise partly from micro-scale processes driven by bacterial symbionts.


One year treatment with omalizumab is effective and well tolerated in Japanese Patients with moderate-to-severe persistent asthma.
We have previously demonstrated that addition of omalizumab to standard therapy improved asthma control by significantly improving lung function and reducing asthma exacerbations in Japanese patients with moderate-to-severe asthma. The aim of this study was to evaluate the effects of omalizumab on long-term disease control in Japanese patients with moderate-to-severe persistent asthma. An open-label, 48-week study was conducted in 133 Japanese patients with moderate-to-severe persistent asthma. Omalizumab was administered subcutaneously every 2 or 4 weeks based on serum IgE level and body weight in each patient. Treatment with omalizumab significantly improved lung function. A subgroup of patients with inadequately controlled severe persistent asthma, despite high dose inhaled corticosteroids and other multiple controller therapies, which corresponds to the Japanese label (label population), showed greater improvements in morning PEF and FEV(1) than the whole study population (full Analysis Set). Serum free IgE levels decreased to below the target and were maintained during the treatment period in almost all patients. The majority of adverse events were mild-to-moderate in severity and there was no trend toward an increase in incidence of adverse events with increase in duration of omalizumab. In addition, the profile of adverse events in this study was similar to that in a 16-week, placebo-controlled study which the present authors had conducted previously in Japan. There were no anaphylactic reactions and no anti-omalizumab antibodies were detected. Long-term treatment with omalizumab is effective and well tolerated in Japanese patients with moderate-to-severe persistent asthma.


Coupling between the mesoscopic dynamics and shear stress of a room-temperature ionic liquid.
Molecular dynamics (MD) simulation is performed on two imidazolium-based ionic liquids, one of which possesses a long alkyl chain whereas the other does not, and the coupling between the liquid structure and the shear stress is examined by means of cross-correlation analysis. The former liquid exhibits a prepeak representing the polar-nonpolar domain structure whereas the latter does not. The relaxation of the intermediate scattering function at the prepeak is slower than those of other microscopic structures. Nonetheless, the principal part of the viscoelastic relaxation is ascribed to the relaxation of the charge-alternation mode in both liquids. In particular, the dynamics of the domain structure hardly seems to affect the viscosity of the ionic liquid with a long alkyl chain, although the cross correlation between the shear stress and the domain mode exists and the relaxation of the cross correlation with the domain mode is far slower than that with the charge alternation mode. The absence of the domain dynamics in the viscoelastic relaxation is ascribed to the decoupling between the relaxations of the short- and the long-range structures.


Characteristics and Hospital Activity of Elderly Patients Receiving Admission Avoidance Home Visits: A Population-Level Record Linkage Study.
As pressures on healthcare systems increase, due to an ageing population, hospital admission avoidance interventions have been emphasised. These interventions can be difficult to objectively evaluate due to non-randomised roll-out, requiring observational methods with carefully selected control groups. This study aims to identify the defining characteristics of elderly patients receiving admission avoidance home visits. We conducted a record linkage study using routinely collected data to compare characteristics and outcomes of the general elderly population and a subset of high-risk patients. Intervention patients were found to have significantly different demographics and admission rates compared to the general population, having four times higher admission rates at baseline. However, they share similarities with high-risk patients, particularly in that after a period of increased admissions, both groups experienced a reduction in the following year. Identifying defining characteristics of the target intervention population can guide the careful selection of a control group for evaluation.


[Doppler sonography findings in arteriitis temporalis].
The results of 145 patients who had been examined by Doppler sonography for suspected giant cell arteritis were analysed. Alternating flow direction, reversed flow direction or "no-flow" in the supratrochlear and/or supraorbital arteries were considered as pathological findings as well as absence of signals from the trunk or branches of one or both superficial temporal arteries. Pathological findings at the periorbital arteries were obtained in 25.9% (n = 7) of the cases with histological proven giant cell arteritis. In a further 29.6% (n = 8) the superficial temporal artery findings were abnormal. Only 13.2% (n = 5) of the patients with normal histological findings and 5.7% (n = 2) of those with degenerative lesions of the temporal arteries had pathological Doppler sonographic findings. Two thirds of the patients with visual impairment showed abnormal findings in the periorbital arteries.


Mutations in genes encoding complement inhibitors CD46 and CFH affect the age at nephritis onset in patients with systemic lupus erythematosus.
Inherited deficiencies of several complement components strongly predispose to systemic lupus erythematosus (SLE) while deficiencies of complement inhibitors are found in kidney diseases such as atypical hemolytic uremic syndrome (aHUS). The exons of complement inhibitor genes CD46 and CFH (factor H) were fully sequenced using the Sanger method in SLE patients with nephritis originating from two cohorts from southern and mid Sweden (n = 196). All identified mutations and polymorphisms were then analyzed in SLE patients without nephritis (n = 326) and in healthy controls (n = 523). We found nonsynonymous, heterozygous mutations in CFH in 6.1% patients with nephritis, in comparison with 4.0% and 5.4% in patients without nephritis and controls, respectively. No associations of SLE or nephritis with common variants in CFH (V62I/Y402H/E936D) were found. Furthermore, we found two nonsynonymous heterozygous mutations in CD46 in SLE patients but not in controls. The A353V polymorphism, known to affect function of CD46, was found in 6.6% of nephritis patients versus 4.9% and 6.1% of the non-nephritis SLE patients and controls. The presence of mutations in CD46 and CFH did not predispose to SLE or nephritis but was associated with earlier onset of nephritis. Furthermore, we found weak indications that there is one protective and one risk haplotype predisposing to nephritis composed of several polymorphisms in noncoding regions of CD46, which were previously implicated in aHUS. SLE nephritis is not associated with frequent mutations in CFH and CD46 as found in aHUS but these may be modifying factors causing earlier onset of nephritis.


Establishing the accuracy of density functional approaches for the description of noncovalent interactions in biomolecules.
Biomolecules have complex structures, and noncovalent interactions are crucial to determine their conformations and functionalities. It is therefore critical to be able to describe them in an accurate but efficient manner in these systems. In this context density functional theory (DFT) could provide a powerful tool to simulate biological matter either directly for relatively simple systems or coupled with classical simulations like the QM/MM (quantum mechanics/molecular mechanics) approach. Additionally, DFT could play a fundamental role to fit the parameters of classical force fields or to train machine learning potentials to perform large scale molecular dynamics simulations of biological systems. Yet, local or semi-local approximations used in DFT cannot describe van der Waals (vdW) interactions, one of the essential noncovalent interactions in biomolecules, since they lack a proper description of long range correlation effects. However, many efficient and reasonably accurate methods are now available for the description of van der Waals interactions within DFT. In this work, we establish the accuracy of several state-of-the-art vdW-aware functionals by considering 275 biomolecules including interacting DNA and RNA bases, peptides and biological inhibitors and compare our results for the energy with highly accurate wavefunction based calculations. Most methods considered here can achieve close to predictive accuracy. In particular, the non-local vdW-DF2 functional is revealed to be the best performer for biomolecules, while among the vdW-corrected DFT methods, uMBD is also recommended as a less accurate but faster alternative.


Expert opinion on tilarginine in the treatment of shock.
Tilarginine is L-N-monomethyl arginine (L-NMMA) or N(G)-monomethyl-L-arginine HCL, a non-selective inhibitor of nitric oxide synthase (NOS), which has been studied in the treatment of septic shock and cardiogenic shock complicating myocardial infarction. Despite strong evidence that excessive nitric oxide (NO) production plays a pivotal role in the pathogenesis of septic shock and may contribute to the pathogenesis of cardiogenic shock complicating myocardial infarction, outcome studies in these two disorders have proved disappointing. L-NMMA therapy was associated with an excess mortality, particularly at doses > 5 mg/(kg h), in septic shock whereas the effects of a lower dose (1 mg/(kg h)) in cardiogenic shock complicating myocardial infarction were neutral. The excess mortality in patients with septic shock was almost certainly the result of unfavourable haemodynamic changes induced by L-NMMA (decreased cardiac output, increased pulmonary vascular resistance and reduced tissue oxygen delivery) whereas the lack of benefit in patients with cardiogenic shock complicating myocardial infarction may have been because the dose of L-NMMA was too low. Further studies of L-NMMA at doses < 5 mg/(kg h) in conjunction with inotrope support may produce more beneficial results. Conversely, the use of a selective inducible NOS inhibitor to reduce the pathological effects of excessive NO production although leaving the beneficial effects of vascular NO production by endothelial NOS unaltered may prove to be of value.


Measurement of cardiac natriuretic hormones (atrial natriuretic peptide, brain natriuretic peptide, and related peptides) in clinical practice: the need for a new generation of immunoassay methods.
Cardiac natriuretic hormones (CNHs) are a family of related peptides, including atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP), and other peptides derived from the N-terminal portion of the proANP and proBNP peptide chains. Assays for cardiac natriuretic peptides have been proposed to help assess clinical conditions associated with expanded fluid volume. In particular, the assays can be useful for distinguishing healthy subjects from patients in different stages of heart failure. Measurements of these hormones have also been considered for prognostic indicators of long-term survival in patients with heart failure and/or after acute myocardial infarction. The different CNHs differ in their production/secretion patterns and have different clearance rates. Furthermore, there are numerous proposed assay configurations for each of these hormones, and it is not clear which assay provides the best pathophysiological and/or clinical information. Here we review recent studies concerning the competitive (such as RIA, enzyme immunoassay, or luminescence immunoassay) and noncompetitive immunoassays (such as two-site IRMA, ELISA, or immunoluminometric assay) for the different cardiac natriuretic peptides to compare the analytical characteristics and clinical relevance of assays for the different CNHs and the different assay formats. Developing sensitive, precise, and accurate immunoassays for cardiac natriuretic peptides has been difficult because of their low concentrations (on average, approximately 3-6 pmol/L) in healthy subjects and because of their structural, metabolic, and physiological characteristics. Competitive assays have historically suffered from lack of sensitivity and specificity for the biologically active peptides. These usually require tedious extraction procedures prior to analysis. Recently, immunometric assays have been developed that have improved sensitivity and specificity; it appears these will be the methods of choice. To date, there is no consensus on the best assay procedure of cardiac natriuretic peptides. To facilitate widespread propagation of determination of these hormones in routine clinical practice, it will be necessary to study the new generation of noncompetitive immunometric methods that are less time-consuming and more sensitive and specific. Although several studies suggest that BNP exhibits better clinical utility than the other CNHs, more studies examining multiple CNHs in the same cohorts of patients will be necessary.


Inflammatory tumor of the cecum simulating neoplasm.
Barium enema performed on a 36-year old man prior to the operative repair of a rectal stricture demonstrated a large cecal filling defect which radiologically was consistent with a villous adenoma or carcinoma. At the time of exploratory celiotomy (for possible ileocolectomy) intraoperative colonoscopy revealed a necrotic appearing lesion in the cecum which did not look neoplastic. Local excision of the mass was therefore performed. The lesion proved to be an inflammatory polyp.


[The mechanisms of the restriction of pollinator range in the fireweed (Chamaenerion angustifolium) and 2 species of geranium (Geranium palustre) and G. pratense)].
Flowers of Chamaenerion angustifolium, Geranium palustre and G. pratense are visited by more than 100 insect species. For all plants the spectrum of visitors is similar. However the role of insects species in pollination is different and depends on the stamen and stigma length, flexibility of pedicle and feeding behaviour of insect inside flower. The possibility to take and to transport pollen grains usually increase with the size of insect. The nature of this correlation is determined by stamen and stigma length. Pollinators of G. palustre with short stamens and stigmas are smaller than those of Ch. angustifolium and G. pratense. On the other hand, more flexible pedicle of G. palustre prevent the flowers from visits of big insects. Three plants studied by the authors are not equally attractive for different insect groups. Dense in fluorescence of Ch. angustifolium and G. pratense that usually are lifter under the grass are very attractive for foraging social insects (honey bee, bumblebee). Flies avoid long distance travelling and prefer single flowers located not far from each other. For instance G. palustre is more attractive for flies not for social bees. It is pollinated mainly by flies and solitary bees with average weight of 10-70 mg. The main pollinators of Ch. angustifolium and G. pratense are honey bees, bumblebees and wasps with average mass exceeding 70 mg.


Phase I radioimmunotherapy of metastatic renal cell carcinoma with 131I-labeled chimeric monoclonal antibody G250.
Clinical tumor targeting studies with chimeric monoclonal antibody G250 (cG250) in renal cell carcinoma (RCC) patients indicated the potential use of this antibody for radioimmunotherapy. Here we report on a phase I activity dose escalation study to determine the safety, the maximum tolerable dose (MTD), and the possible therapeutic potential of 131I-labeled cG250 in patients with progressive metastatic RCC. All patients (n = 12) received a diagnostic i.v. infusion of 5 mg of cG250 labeled with 222 MBq of 131I. If accumulation of the antibody in metastatic lesions was observed, patients were hospitalized and a second, therapeutic, i.v. infusion of 5 mg of cG250 labeled with a high dose of 131I was administered (n = 8). Three patients per dose level were entered, starting at 1665 MBq/m2. If no dose-limiting toxicity occurred, the study continued at the next dose level (555 MBq/m2 increase). Most patients experienced mild nausea without vomiting. No other complaints were reported during hospitalization. In two of two patients who received a dose of 2775 MBq/m2, grade IV hematological toxicity was observed, which was defined as dose limiting. Thus, the MTD was set at 2220 MBq/m2. In one patient (2220 MBq/m2), stable disease (lasting 3-6 months) was achieved, whereas another patient (2220 MBq/m2) showed a partial response that is ongoing (>9 months). The minor responses observed in this phase I trial in patients with an advanced stage of RCC are encouraging and warrant further study in a phase II setting at the MTD to determine the efficacy of radioimmunotherapy for metastatic RCC.


[Construction of the synthetic genes for protein analogs of spider silk carcass spidroin 1 and their expression in tobacco plants].
To obtain transgenic tobacco plants expressing recombinant analogs of spider dragline silk spidroin 1, artificial 1f5 and 1f9 coding for spidroin 1 analogs were 3'-fused in-frame with the reporter lichenase gene. The Tr2' weak constitutive promoter of Agrobacterium tumefaciens T-DNA and the strong constitutive promoter of the cauliflower mosaic virus 35S RNA gene were used as regulatory elements. The expression cassettes were used to transform agrobacteria and then introduced in tobacco leaf disks. On evidence of Southern hybridization, transgenic plants each carried a single copy of a hybrid gene, which corresponded in size to the constructed one. Zymography and Western blotting revealed full-length hybrid proteins in leaf extracts of transgenic plants. The results testified that plants can maintain and express synthetic genes for spider silks and, consequently, may be used as a convenient producer of recombinant silk analogs.


The dynamics of attentional sampling during visual search revealed by Fourier analysis of periodic noise interference.
What are the temporal dynamics of perceptual sampling during visual search tasks, and how do they differ between a difficult (or inefficient) and an easy (or efficient) task? Does attention focus intermittently on the stimuli, or are the stimuli processed continuously over time? We addressed these questions by way of a new paradigm using periodic fluctuations of stimulus information during a difficult (color-orientation conjunction) and an easy (+ among Ls) search task. On each stimulus, we applied a dynamic visual noise that oscillated at a given frequency (2-20 Hz, 2-Hz steps) and phase (four cardinal phase angles) for 500 ms. We estimated the dynamics of attentional sampling by computing an inverse Fourier transform on subjects' d-primes. In both tasks, the sampling function presented a significant peak at 2 Hz; we showed that this peak could be explained by nonperiodic search strategies such as increased sensitivity to stimulus onset and offset. Specifically in the difficult task, however, a second, higher-frequency peak was observed at 9 to 10 Hz, with a similar phase for all subjects; this isolated frequency component necessarily entails oscillatory attentional dynamics. In a second experiment, we presented difficult search arrays with dynamic noise that was modulated by the previously obtained grand-average attention sampling function or by its converse function (in both cases omitting the 2 Hz component to focus on genuine oscillatory dynamics). We verified that performance was higher in the latter than in the former case, even for subjects who had not participated in the first experiment. This study supports the idea of a periodic sampling of attention during a difficult search task. Although further experiments will be needed to extend these findings to other search tasks, the present report validates the usefulness of this novel paradigm for measuring the temporal dynamics of attention.


Male GPs' views on men seeking medical help: a qualitative study.
Analysis of health service statistics reveals that men tend to under-use primary health care services. Previous research has suggested that men view illness as a form of weakness and have negative views about men who seek medical help. It has also indicated that some men believe that male General Practitioner (GP)s hold similar views, a factor which may influence their own tendency to self-refer. This study aims to explore male GPs' experiences of their male patients' patterns of self-referral and to identify factors that may negatively influence doctor-patient relationships. Semi-structured interviews were carried out with 10 male GPs (ages=35-53). These were audiotaped and subjected to interpretative phenomenological analysis to identify relevant themes. For all participants, managing service demands were seen as a major challenge. Analysis of these GP's accounts suggested that they see men's self-referral as being primarily influenced by the need to conform to masculine gender roles and a number of ways that men accessed services in a way congruent with this were identified. This study also explored how participants' own adherence to gender roles may itself influence the doctor-patient relationship. This study suggests that, like other men, male GPs may have ambivalent attitudes towards male self-referral and that this may influence their interactions with male patients.


[Patient-Centered Care model and New Public Management : Confluence and paradox.]
The search for efficiency and concentration on patient needs and satisfaction are at the topic of current conceptions in terms of the quality and organization of health services. There is indeed a relative consolidation of organizational approaches called Patient-Centered Care (e.g. CCM, DM, case management, population management) for the structured management of patients with chronic diseases and a real desire to implement them in the domain of health. This evolution continues in the management of health systems through New Public Management - NPM. There is a strong pressure on professionals (care professionals and managers) to ensure greater efficiency in terms of activities and greater efficiency in terms of the use of resources. Both developments point to a greater emphasis on personalizing care, differentiating benefits, and taking into account the satisfaction of beneficiaries of public services. However, without being antinomic, the two evolutions underlie divergent and even paradoxical issues and mechanisms.


Are genital examinations necessary for STI screening for female sex workers? An audit of decriminalized and regulated sex workers in Melbourne, Australia.
The Victorian legislation prohibits sex workers from working when they have visible anogenital herpes or warts. The aim of this study was to determine the proportion of asymptomatic female sex workers (FSW) diagnosed with anogenital herpes or warts by genital examination. We analysed all computerised medical records of consultations with FSW at the Melbourne Sexual Health Centre (MSHC) in 2018. All asymptomatic sex workers were offered screening sexually transmitted infections (STIs) and a genital examination to identify visible anogenital herpes or warts at MSHC. FSW consultations were categorised into either 'asymptomatic' or 'symptomatic' based on the presence of symptoms reported by the FSW to the triage nurse. The proportion of asymptomatic FSW diagnosed with visible anogenital herpes or warts during a routine screening examination was calculated. In 2018, 4055 consultations were provided to 1979 FSW. 3406 of these consultations were asymptomatic and all were examined by an experienced clinician for signs of STIs. Of these 3406 asymptomatic consultations, seven FSW (0.21%, 95% CI: 0.08% to 0.42%) were diagnosed with visible anogenital herpes and/or warts following a genital examination. Four were diagnosed with warts (0.12%, 95% CI: 0.03% to 0.30%), two with herpes (0.06%, 95% CI: 0.01% to 0.21%) and one with both herpes and warts (0.03%, 95% CI: 0.001% to 0.16%). Based on these data, approximately 500 asymptomatic FSW would need to be examined to identify one case of anogenital herpes or warts. Genital examinations consume considerable clinical resources, increase the duration of consultations and provide essentially no significant benefit to the mandated testing for gonorrhoea, chlamydia, HIV and syphilis. Our clinic will use self-collected samples and no longer examine FSW who are asymptomatic.


[Living and working conditions and access to health services for agricultural and non-agricultural workers, Brazil, 2013].
In Brazil, the growth of agribusiness to the detriment of family agriculture occurred while concealing social, environmental and human health damages. The objective was to compare living and working conditions and access to health services between agricultural and non-agricultural workers. Data from the National Health Survey (PNS) on living and working conditions, sociodemographic, economic characteristics, and access to health services from a representative sample of the employed Brazilian population were adopted. Pearson's chi-square test was used, with a significance level of 0.05, taking the complex sampling design into consideration. Agricultural workers suffered from worse living conditions, lower purchasing power, greater exposure to solar radiation and chemical agents, and a higher frequency and severity of occupational accidents compared to non-agricultural workers. The agricultural population had greater coverage of the Family Health Service and sought medical care from the Unified Health System (SUS) to treat diseases, while the non-agricultural workers sought private medical care for preventive actions. The differences found between these workers imply different patterns of illness and define specific health needs.


Urinary bladder smooth muscle regeneration utilizing bone marrow derived mesenchymal stem cell seeded elastomeric poly(1,8-octanediol-co-citrate) based thin films.
Bladder regeneration studies have yielded inconclusive results possibly due to the use of unfavorable cells and primitive scaffold design. We hypothesized that human mesenchymal stem cells seeded onto poly(1,8-octanediol-co-citrate) elastomeric thin films would provide a suitable milieu for partial bladder regeneration. POCfs were created by reacting citric acid with 1,8-octanediol and seeded on opposing faces with human MSCs and urothelial cells; normal bladder smooth muscle cells and UCs, or unseeded POCfs. Partial cystectomized nude rats were augmented with the aforementioned POCfs, enveloped with omentum and sacrificed at 4 and 10 weeks. Isolated bladders were subjected to Trichrome and anti-human gamma-tubulin, calponin, caldesmon, smooth muscle gamma-actin, and elastin stainings. Mechanical testing of POCfs revealed a Young's modulus of 138 kPa with elongation 137% its initial length without permanent deformation demonstrating its high uniaxial elastic potential. Trichrome and immunofluorescent staining of MSC/UC POCf augmented bladders exhibited typical bladder architecture with muscle bundle formation and the expression and retention of bladder smooth muscle contractile proteins of human derivation. Quantitative morphometry of MSC/UC samples revealed muscle/collagen ratios approximately 1.75x greater than SMC/UC controls at 10 weeks. Data demonstrate MSC seeded POCfs support partial regeneration of bladder tissue in vivo.


Pulmonary paraganglioma manifesting as an endobronchial mass.
Thoracic paragangliomas comprise only 1-2% of all paragangliomas, including the adrenal pheochromocytomas, and these tumors are mostly found in the mediastinal compartments (1). To the best of our knowledge, there is only one case report in the pathology literature of endobronchial involvement by a primary pulmonary paraganglioma (2). We report here on the CT and bronchoscopic findings of a case of pathologically proven endobronchial paraganglioma in a 37-year-old woman. In our case, bronchoscopy and CT demonstrated an endobronchial hypervascular mass, which indicated the presence of carcinoid or hypervascular metastasis based on the known incidence of such tumors.


[Changes in the configuration of synapses in the cerebral cortex in laboratory rats after termination of epileptic seizures during kindling].
The sensorimotor area of the cerebral cortex of the laboratory rat was repeatedly stimulated by electric current after 10-minute intervals which led to the development a gradual prolongation of self-sustained after-discharges (SSAD). One and 10 minutes after termination of the third SSAD the authors analyzed under the electron microscope synapses type I after to Gray from the second cortical homotopic area of the unstimulated hemisphere. They evaluated the curvature of the active zone of synapses. Evidence was provided that one minute after termination of the fit the number of negatively curved synapses increased and the number of synapses with a straight active zone decreased. The number of synapses with a positively curved active zone does not change. Ten minutes after termination of SSAD in the experimental animals the number of positively curved elements declines and the number of negatively curved synapses increases. There is no significant difference in the number of synapses with a straight active zone. The authors discuss possible functional consequences of the mentioned changes.


Complexities surrounding the diagnosis and management of hypercalcaemia in pregnancy.
Hypercalcaemia in pregnancy is uncommon, with associated adverse obstetric and perinatal outcomes for both the mother and the fetus. Determination of causality is central to its management. Diagnostic imaging techniques are limited during pregnancy and the diagnosis is made more complex by physiological changes in calcium and vitamin D homeostasis in pregnancy. Further, therapeutic options are limited due to safety considerations for the pregnant woman and the developing foetus. Three cases of hypercalcaemia in pregnancy will be presented, highlighting the distinct aetiologies and management strategies for hypercalcaemia in pregnancy and the importance of early measurement of serum calcium in pregnancy screening. There are complex physiological changes in calcium balance in pregnancy, including increased calcium intestinal absorption and renal excretion. Hypercalcaemia in pregnancy is uncommon but has important potential maternal and foetal complications, making a compelling argument for routine antenatal, calcium screening. Identifying the cause of hypercalcaemia in pregnancy can be challenging due to the complex placental interplay in biochemical test interpretation and due to safety constraints restricting imaging and surgery. Acute medical management of hypercalcaemia must be considered in the context of both maternal and foetal well-being, along with gestational age and specific consideration for the safety of the developing fetus in late gestation.


Induction of NEIL1 and NEIL2 DNA glycosylases in aniline-induced splenic toxicity.
The mechanisms by which aniline exposure elicits splenotoxic response, especially the tumorigenic response, are not well-understood. Earlier, we have shown that aniline-induced oxidative stress is associated with increased oxidative DNA damage in rat spleen. The base excision repair (BER) pathway is the major mechanism for the repair of oxidative DNA base lesions, and we have shown an up-regulation of 8-oxoguanine glycosylase 1 (OGG1), a specific DNA glycosylase involved in the removal of 8-hydroxy-2'-deoxyguanosine (8-OHdG) adducts, following aniline exposure. Nei-like DNA glycosylases (NEIL1/2) belong to a family of BER proteins that are distinct from other DNA glycosylases, including OGG1. However, contribution of NEIL1/2 in the repair of aniline-induced oxidative DNA damage in the spleen is not known. This study was, therefore, focused on evaluating if NEILs also contribute to the repair of oxidative DNA lesions in the spleen following aniline exposure. To achieve that, male SD rats were subchronically exposed to aniline (0.5 mmol/kg/day via drinking water for 30 days), while controls received drinking water only. The BER activity of NEIL1/2 was assayed using a bubble structure substrate containing 5-OHU (preferred substrates for NEIL1 and NEIL2) and by quantitating the cleavage products. Aniline treatment led to a 1.25-fold increase in the NEIL1/2-associated BER activity in the nuclear extracts of spleen compared to the controls. Real-time PCR analysis for NEIL1 and NEIL2 mRNA expression in the spleen revealed 2.7- and 3.9-fold increases, respectively, in aniline-treated rats compared to controls. Likewise, Western blot analysis showed that protein expression of NEIL1 and NEIL2 in the nuclear extract of spleens from aniline-treated rats was 2.0- and 3.8-fold higher than controls, respectively. Aniline treatment also led to stronger immunoreactivity for NEIL1 and NEIL2 in the spleens, confined to the red pulp areas. These studies, thus, show that aniline-induced oxidative stress is associated with an induction of NEIL1/2. The increased NIEL-mediated BER activity is another indication of aniline-induced oxidative damage in the spleen and could constitute another important mechanism of removal of oxidative DNA lesions, especially in transcribed DNA following aniline insult.


Surgical Treatment of Erectile Dysfunction and Peyronie's Disease Using Malleable Prosthesis.
Peyronie's disease (PD) is a condition of middle aged men and frequently accompanied by erectile dysfunction (ED) which was attributed to penile deformity, vascular pathology and psychological components. The implantation of semi-rigid penile prosthesis allows for these patients to undergo a simple procedure aimed at correction both penile deformity and ED. The aim of this study was to investigate surgical and clinical outcomes and patient satisfaction rate at long term follow-up after semi-rigid penile prosthesis implantation (PPI) in men with PD and ED. A total of 66 patients with mean age of 49.2 (range, 30-76) years old underwent semi-rigid PPI between 1995 and 2006. Genesis (Coloplast®) was used for implantation in a standard manner by penoscrotal approach without using any graft and remodeling technique. In all patients, dilatation of corpora was performed without any difficulty and straightening of the penis was achieved. A retrospective review of clinical database and prospective telephone survey were conducted in all patients. The mean follow-up time was 9.7 years (range, 6 to 17). There wasn't any clinical infection and complication during follow-up period. Fifty-nine patients were sexually active at the time of the interview. None of the patients reported residual curvature. The overall patient satisfaction was 91.5% (54 patients). Primary reasons for dissatisfaction were decreased penile length and prosthesis problems. Based on our results semi-rigid PPI is effective and easy procedure for treatment of men with PD and ED without any complication and with high patient satisfaction rate in long-term follow up period.


Composite poly(ethylene oxide)-based solid electrolyte with consecutive and fast ion transport channels constructed by upper-dimensional MIL-53(Al) nanofibers.
Novel structural designs for metal organic frameworks (MOFs) are expected to improve ion-transport behavior in composite solid electrolytes. Herein, upper-dimensional MIL-53(Al) nanofibers (MNFs, MIL-53 belongs to the MIL (Material Institute Lavoisier) group) with flower-like nanoflake structures have been designed and constructed via modified hydrothermal coordination. The optimized MNFs with high surface area and porosity can form abundant interfaces with poly(ethylene oxide) (PEO) matrix. The plasticization of MNFs to the PEO matrix will facilitate segmental movement of PEO chains to facilitate Li+ conduction. The unsaturated open metal centers of MNFs can effectively capture bis(trifluoromethanesulfonyl)imide anions (TFSI-) to deliver more free lithium ions for transfer. Moreover, the upper-dimensional nanofiber structure endows lithium ions with a long-range and consecutive transport pathway. The obtained composite solid electrolyte (MNFs@PEO) presents a high ionic conductivity of 4.1 × 10-4 S cm-1 and a great Li+ transference number of 0.4 at 60 °C. The electrolyte also exhibits a stable Li plating/stripping behavior over 1000 h at 0.1 mA cm-1 with inhibited Li dendrite growth. Furthermore, the Li/LiFePO4 and Li/LiNi0.8Mn0.1Co0.1O2 batteries with MNFs@PEO as electrolytes both display great cycling stabilities with high-capacity retention, indicating their potential applications in lithium metal batteries. The study will put forward new inspirations for designing advanced MOF-based composite solid electrolytes.


The nieuwkoid/dharma homeobox gene is essential for bmp2b repression in the zebrafish pregastrula.
Dorsoventral specification of the zebrafish gastrula is governed by the functions of the dorsal shield, a region of the embryo functionally analogous to the amphibian Spemann organizer. We report that the bozozok locus encodes the transcription factor nieuwkoid/dharma, a homeobox gene with non-cell-autonomous organizer-inducing activity. The nieuwkoid/dharma gene is expressed prior to the onset of gastrulation in a restricted region of an extraembryonic tissue, the yolk syncytial layer, that directly underlies the presumptive organizer cells. A single base-pair substitution in the nieuwkoid/dharma gene results in a premature stop codon in boz(m168) mutants, leading to the generation of a truncated protein product which lacks the homeodomain and fails to induce a functional organizer in misexpression assays. Embryos homozygous for the boz(m168) mutation exhibit impaired dorsal shield specification often leading to the loss of shield derivatives, such as prechordal plate in the anterior and notochord in the posterior, along the entire anteroposterior axis. Furthermore, boz homozygotes feature a loss of neural fates anterior to the midbrain/hindbrain boundary. Characterization of homozygous mutant embryos using molecular markers indicates that the boz ventralized phenotype may be due, in part, to the derepression of a secreted antagonizer of dorsal fates, zbmp2b, on the dorsal side of the embryo prior to the onset of gastrulation. Furthermore, ectopic expression of nieuwkoid/dharma RNA is sufficient to lead to the down regulation of zbmp2b expression in the pregastrula. Based on these results, we propose that gastrula organizer specification requires the Nieuwkoop center-like activity mediated by the nieuwkoid/dharma/bozozok homeobox gene and that this activity reveals the role of a much earlier than previously suspected inhibition of ventral determinants prior to dorsal shield formation.


An observed correlation between plume activity and tidal stresses on Enceladus.
Saturn's moon Enceladus emits a plume of water vapour and micrometre-sized ice particles from a series of warm fissures located near its south pole. This geological activity could be powered or controlled by variations in the tidal stresses experienced by Enceladus as it moves around its slightly eccentric orbit. The specific mechanisms by which these varying stresses are converted into heat, however, are still being debated. Furthermore, it has proved difficult to find a clear correlation between the predicted tidal forces and measured temporal variations in the plume's gas content or the particle flux from individual sources. Here we report that the plume's horizontally integrated brightness is several times greater when Enceladus is near the point in its eccentric orbit where it is furthest from Saturn (apocentre) than it is when near the point of closest approach to the planet (pericentre). More material therefore seems to be escaping from beneath Enceladus' surface at times when geophysical models predict its fissures should be under tension and therefore may be wider open.


Kainic Acid lesion of vasopressinergic neurons in the hypothalamus disrupts flank marking behavior in golden hamsters.
Abstract It is well established that flank marking behavior in the Golden hamster is controlled by vasopressin-sensitive neurons localized to the anterior hypothalamus; however, the source(s) of vasopressinergic innervation to this area is unknown. Previous analysis by immunocytochemistry showed distinct populations of vasopressinergic magnocellular neurons localized to the supraoptic nucleus, paraventricular nucleus and the nucleus circularis that did not project to the neurohypophysis. In the present study, these same hypothalamic nuclei were lesioned by microinjection of kainic acid to determine which, if any, of these populations of vasopressin neurons are involved in the control of flank marking. Unilateral lesions in the areas of the nucleus circularis and supraoptic nucleus at the rostro-caudal plane of the anterior hypothalamus abolished odor-induced flank marking behavior. Lesions in the paraventricular nucleus at the level of the anterior hypothalamus did not consistently inhibit flank marking, while lesions of magnocellular neurons rostral or caudal to the anterior hypothalamus were ineffective. The microinjection of vasopressin into the anterior hypothalamus following lesion of the paraventricular and supraoptic nuclei stimulated flank marking, evidence that treatment with kainic acid did not damage the efferent component of this behavior. However, animals with lesions in the nucleus circularis did not respond to the microinjection of vasopressin; hence, it is uncertain whether lesions in this area disrupt vasopressinergic innervation to the anterior hypothalamus or simply destroy the motor neurons controlling flank marking. In summary, the data clearly demonstrate that vasopressin neurons localized primarily to the medial aspect of the supraoptic nucleus are necessary for sensory integration of odor-induced flank marking, and as such, may be one possible source of neurotransmitter controlling this behavior.


Cardiac contractility modulation improves long-term survival and hospitalizations in heart failure with reduced ejection fraction.
Cardiac contractility modulation (CCM) improves symptoms and exercise tolerance and reduces heart failure (HF) hospitalizations over 6-month follow-up in patients with New York Heart Association (NYHA) class III or IV symptoms, QRS < 130 ms and 25% ≤ left ventricular ejection fraction (LVEF) ≤ 45% (FIX-HF-5C study). The current prospective registry study (CCM-REG) aimed to assess the longer-term impact of CCM on hospitalizations and mortality in real-world experience in this same population. A total of 140 patients with 25% ≤ LVEF ≤ 45% receiving CCM therapy (CCM-REG25-45 ) for clinical indications were included. Cardiovascular and HF hospitalizations, Minnesota Living with Heart Failure Questionnaire (MLHFQ) and NYHA class were assessed over 2 years. Mortality was tracked through 3 years and compared with predictions by the Seattle Heart Failure Model (SHFM). A separate analysis was performed on patients with 35% ≤ LVEF ≤ 45% (CCM-REG35-45 ) and 25% ≤ LVEF < 35% (CCM-REG25-34 ). Hospitalizations decreased by 75% (from 1.2/patient-year the year before, to 0.35/patient-year during the 2 years following CCM, P < 0.0001) in CCM-REG25-45 and by a similar amount in CCM-REG35-45 (P < 0.0001) and CCM-REG25-34 . MLHFQ and NYHA class improved in all three cohorts, with progressive improvements over time (P < 0.002). Three-year survival in CCM-REG25-45 (82.8%) and CCM-REG24-34 (79.4%) were similar to those predicted by SHFM (76.7%, P = 0.16; 78.0%, P = 0.81, respectively) and was better than predicted in CCM-REG35-45 (88.0% vs. 74.7%, P = 0.046). In real-world experience, CCM produces results similar to those of previous studies in subjects with 25% ≤ LVEF ≤ 45% and QRS < 130 ms; cardiovascular and HF hospitalizations are reduced and MLHFQ and NYHA class are improved. Overall mortality was comparable to that predicted by the SHFM but was lower than predicted in patients with 35% ≤ LVEF ≤ 45%.


Entomological studies at an enzootic Venezuelan equine encephalitis virus focus in Guatemala, 1977-1980.
The ecology of several potential mosquito vectors of Venezuelan equine encephalitis (VEE) alphavirus was studied in an enzootic focus of that virus on the Pacific coast of Guatemala over a four-year period. Four species-Culex taeniopus, Mansonia titillans, Culex nigripalpus and Aedes taeniorhynchus-were most prevalent during the wet season when transmission normally occurs. However, only Cx. taeniopus yielded VEE virus. The bloodfeeding patterns of these species revealed that Ae. taeniorhynchus and Ma. titillans fed almost exclusively on bovine and equine hosts. Conversely, Cx. nigripalpus was highly ornithophilic but occasionally fed on mammals. Cx. taeniopus exhibited a wide host range, utilizing both large and small mammals as well as birds and, rarely, reptiles. The versatility in feeding pattern displayed by this mosquito coupled with its ability to become infected with relatively low levels of enzootic VEE virus suggests that vertebrates other than rodents may serve as amplifying hosts in this habitat. Nepuyo virus was also isolated from Cx. taeniopus, suggesting that this mosquito might be an endemic vector of this rodent-associated bunyavirus. A single isolate of St. Louis encephalitis virus was made from Cx. nigripalpus.