Homozygous loss-of-function variants in European cosmopolitan and isolate populations.
Homozygous loss of function (HLOF) variants provide a valuable window on gene function in humans, as well as an inventory of the human genes that are not essential for survival and reproduction. All humans carry at least a few HLOF variants, but the exact number of inactivated genes that can be tolerated is currently unknown—as are the phenotypic effects of losing function for most human genes. Here, we make use of 1432 whole exome sequences from five European populations to expand the catalogue of known human HLOF mutations; after stringent filtering of variants in our dataset, we identify a total of 173 HLOF mutations, 76 (44%) of which have not been observed previously. We find that population isolates are particularly well suited to surveys of novel HLOF genes because individuals in such populations carry extensive runs of homozygosity, which we show are enriched for novel, rare HLOF variants. Further, we make use of extensive phenotypic data to show that most HLOFs, ascertained in population-based samples, appear to have little detectable effect on the phenotype. On the contrary, we document several genes directly implicated in disease that seem to tolerate HLOF variants. Overall HLOF genes are enriched for olfactory receptor function and are expressed in testes more often than expected, consistent with reduced purifying selection and incipient pseudogenisation.


Moving for medicaid? Recent eligibility expansions did not induce migration from other states.
Starting in 2014, many low-income adult residents of states that forgo the Affordable Care Act's expansion of Medicaid would be eligible for that program if they moved to a state that had chosen to expand its coverage. Some of these people may migrate to receive coverage, thereby increasing costs for states that have expanded the program. This is known as the "welfare magnet" hypothesis, a claim that geographic variation in social programs induces the migration of welfare recipients to places with more generous benefits or eligibility. To investigate whether such spillover effects are likely, we used data from the Current Population Survey to examine the migration patterns of low-income people before and after recent expansions of public insurance in Arizona, Maine, Massachusetts, and New York. Using difference-in-differences analysis of migration in expansion and control states, we found no evidence of significant migration effects. Our preferred estimate was precise enough to rule out net migration effects of larger than 1,600 people per year in an expansion state. These results suggest that migration will not be a common way for people to obtain Medicaid coverage under the current expansion and that interstate migration is not likely to be a significant source of costs for states choosing to expand their programs.


Biosensor assay for determination of haptoglobin in bovine milk.
Despite more than 30 years of research into mastitis diagnostics, there are few alternatives to the somatic cell count (SCC) in practical use for identification of cows with subclinical mastitis. Mastitis is not only an animal welfare problem, but also affects the yield, composition and technological properties of milk. Hence, dairy cooperatives give farmers a premium quality payment to encourage low SCC although there is no clear scientific data defining the level of SCC in bulk tank milk that is associated with additional benefits in terms of milk quality. Recent research on alternative markers for inflammatory reactions in the lactating cow, e.g. in mastitis, includes investigations of the acute phase protein, haptoglobin (Hp). So far, the content of Hp in milk has mainly been studied in relation to mastitis diagnostics, with little attention given to its importance for milk composition and technological properties. At present, Hp in milk is measured using ELISA, but this technique is not suitable for routine large-scale analysis. In recent years, optical biosensor technology has been used for automated and rapid quantitative analysis of different components in milk, but so far not for analysis of acute phase proteins. The aim of the present study was to develop a rapid and sensitive biosensor method to determine Hp in milk. An affinity sensor assay based on the interaction between Hp and haemoglobin was developed using surface plasmon resonance (SPR) biosensor technology. The assay was used to analyse Hp in composite milk samples from cows without any clinical signs of mastitis and quarter milk samples with a weak to strong reaction in the California Mastitis Test (CMT). A commercial ELISA for determination of Hp in milk was used for comparison. The limit of detection (LOD) of the biosensor assay was determined as 1.1 mg/l. Within-assay and between-day variations were determined both with bulk tank milk spiked with human Hp and with composite milk samples containing bovine Hp. Coefficients of variation varied between 3.6 and 8.6% at concentrations between 4.0 and 12 mg/l, respectively. Agreement between the results obtained by the biosensor assay and the ELISA was satisfactory; however, the results obtained by the biosensor were generally lower than the results obtained by the ELISA. Possible explanations for this observation are discussed.


Fragility Fracture Risk in Elderly Patients With Cervical Myelopathy.
Retrospective review. To identify and compare the incidences of fragility fractures amongst three elderly populations: the general population, patients with surgically treated cervical spondylotic myelopathy (CSM), and patients with CSM not surgically treated. CSM is a common disease in the elderly. Progression of myelopathic symptoms, including gait imbalance, can be a source of morbidity as it can lead to increased falls. Records of elderly patients with Medicare insurance from 2005 to 2014 were retrospectively reviewed. Three mutually exclusive populations of patients were identified for analysis, including a cohort of patients with a diagnosis of CSM who were not treated with surgery; a cohort of patients with CSM who were treated with surgery; and a group of control patients who had never been treated with cervical spine surgery nor were diagnosed with CSM. Incidence of fractures of the distal radius, proximal humerus, proximal femur, and lumbar spine were assessed and compared between cohorts, adjusted by age, sex, osteoporosis, dementia, cerebrovascular disease, and Charlson Comorbidity Index. A total of 891,864 patients were identified, of which 60,332 had a diagnosis of CSM and 24,439 underwent cervical spine surgery. Compared to general population controls, the 12-month adjusted odds of experiencing at least one fragility fracture were 1.59 times higher in patients with CSM who were not treated with surgery (P < 0.001). The analogous odds ratio was 1.34 (P < 0.001) at 3 years. Compared to nonsurgically treated patients with CSM, the odds of experiencing at least one fragility fracture were reduced to 0.89 in surgically treated patients (P = 0.008). Fragility fractures are a significant source of morbidity and mortality in elderly patients. CSM is associated with increased rates of fragility fractures, although surgical management of CSM may be protective against risk of fragility fracture. 3.


Flaw tolerance of nuclear intermediate filament lamina under extreme mechanical deformation.
The nuclear lamina, composed of intermediate filaments, is a structural protein meshwork at the nuclear membrane that protects genetic material and regulates gene expression. Here we uncover the physical basis of the material design of nuclear lamina that enables it to withstand extreme mechanical deformation of >100% strain despite the presence of structural defects. Through a simple in silico model we demonstrate that this is due to nanoscale mechanisms including protein unfolding, alpha-to-beta transition, and sliding, resulting in a characteristic nonlinear force-extension curve. At the larger microscale this leads to an extreme delocalization of mechanical energy dissipation, preventing catastrophic crack propagation. Yet, when catastrophic failure occurs under extreme loading, individual protein filaments are sacrificed rather than the entire meshwork. This mechanism is theoretically explained by a characteristic change of the tangent stress-strain hardening exponent under increasing strain. Our results elucidate the large extensibility of the nuclear lamina within muscle or skin tissue and potentially many other protein materials that are exposed to extreme mechanical conditions, and provide a new paradigm toward the de novo design of protein materials by engineering the nonlinear stress-strain response to facilitate flaw-tolerant behavior.


Hip fractures in persons with stroke.
Background. Our aim was to determine the incidence of hip fractures within two years after stroke, to identify associated factors, to evaluate which test instruments that best could identify people at risk, and to describe the circumstances that prevailed when they sustained their hip fractures. Method. A total of 377 persons with first-ever stroke were followed up for a 24-month period. Stroke severity, cognition, and associated medical conditions were registered. The following test instruments were used: National Institutes of Health Stroke Scale, Mini-Mental State Examination, Berg Balance Scale, Timed Up & Go, and Stops Walking When Talking. Result. Sixteen of the persons fractured their hip within the study period, which corresponds to an incidence of 32 hip fractures per 1000 person-years. Persons with fractures more often had impaired vision and cognitive impairment and more had had previous fractures. Of the investigated test instruments, Timed Up & Go was the best test to predict fractures. Conclusion. The incidence of hip fractures in persons with stroke was high in this study. Persons with previous fractures, and visual and cognitive defects are at the greatest risk. Certain test instruments could be used in order to find people at risk, which should be targeted for fall preventive measures.


Clinical guide for audiologic tinnitus management II: Treatment.
This article is the second of 2 that address the need for basic procedures that can be used commonly by audiologists to manage patients with clinically significant tinnitus, as well as hyperacusis. The method described is termed audiologic tinnitus management (ATM). ATM was developed specifically for use by audiologists. Although certain procedural components were adapted from the methods of tinnitus masking and tinnitus retraining therapy, ATM is uniquely and specifically defined. A detailed description of the ATM assessment procedures is provided in the companion article (J. A. Henry, T. L. Zaugg, & M. A. Schechter, 2005). The present article describes a specific clinical protocol for providing treatment with ATM. The treatment method described for ATM includes structured informational counseling and an individualized program of sound enhancement that can include the use of hearing aids, ear-level noise generators, combination instruments (noise generator and hearing aid combined), personal listening devices (wearable CD, tape, and MP3 players), and augmentative sound devices (e.g., tabletop sound generators). Ongoing treatment appointments involve primarily the structured counseling, evaluation, and adjustment of the use of sound devices, and assessment of treatment outcomes. The informational counseling protocol and an interview form for determining treatment outcomes are each described in step-by-step detail for direct clinical application. This article can serve as a practical clinical guide for audiologists to provide treatment for tinnitus in a uniform manner.


The probiotic Lactobacillus coryniformis CECT5711 reduces the vascular pro-oxidant and pro-inflammatory status in obese mice.
Obesity is associated with intestine dysbiosis and is characterized by a low-grade inflammatory status, which affects vascular function. In the present study, we evaluated the effects of a probiotic with immunomodulatory properties, Lactobacillus coryniformis CECT5711, in obese mice fed on an HFD (high-fat diet). The probiotic treatment was given for 12 weeks, and it did not affect the weight evolution, although it reduced basal glycaemia and insulin resistance. L. coryniformis administration to HFD-induced obese mice induced marked changes in microbiota composition and reduced the metabolic endotoxaemia as it decreased the LPS (lipopolysaccharide) plasma level, which was associated with a significant improvement in gut barrier disruption. Furthermore, it lowered TNFα (tumour necrosis factor α) expression in liver, improving the inflammatory status, and thus the glucose metabolism. Additionally, the probiotic reversed the endothelial dysfunction observed in obese mice when endothelium- and NO (nitric oxide)-dependent vasodilatation induced by acetylcholine in aortic rings was studied. It also restored the increased vessel superoxide levels observed in obese mice, by reducing NADPH oxidase activity and increasing antioxidant enzymes. Moreover, chronic probiotic administration for 2 weeks also improved endothelial dysfunction and vascular oxidative stress induced by in vivo administration of LPS in control mice fed on a standard chow diet. The results of the present study demonstrate an endothelial-protective effect of L. coryniformis CECT5711 in obese mice by increasing NO bioavailability, suggesting the therapeutic potential of this gut microbiota manipulation to prevent vasculopathy in obesity.


Assemblathon 1: a competitive assessment of de novo short read assembly methods.
Low-cost short read sequencing technology has revolutionized genomics, though it is only just becoming practical for the high-quality de novo assembly of a novel large genome. We describe the Assemblathon 1 competition, which aimed to comprehensively assess the state of the art in de novo assembly methods when applied to current sequencing technologies. In a collaborative effort, teams were asked to assemble a simulated Illumina HiSeq data set of an unknown, simulated diploid genome. A total of 41 assemblies from 17 different groups were received. Novel haplotype aware assessments of coverage, contiguity, structure, base calling, and copy number were made. We establish that within this benchmark: (1) It is possible to assemble the genome to a high level of coverage and accuracy, and that (2) large differences exist between the assemblies, suggesting room for further improvements in current methods. The simulated benchmark, including the correct answer, the assemblies, and the code that was used to evaluate the assemblies is now public and freely available from http://www.assemblathon.org/.


Immunoelectron microscopic studies of the sites of cell-substratum and cell-cell contacts in cultured fibroblasts.
Our object was to obtain information about the molecular structures present at cell-substratum and cell-cell contact sites formed by cultured fibroblasts. We have carried out double immunoelectron-microscopic labeling experiments on ultrathin frozen sections cut through such contact sites to determine the absolute and relative dispositions of the three proteins fibronectin, vinculin, and alpha-actinin with respect to these sites. (a) Three types of cell-substratum and cell-cell contact sites familiar from plastic sections could also be discriminated in the frozen sections by morphological criteria alone, i.e., the gap distances between the two surfaces, and the presence of submembranous densities. These types were: (i) focal adhesions (FA); (ii) close contacts (CC); and (iii) extracellular matrix contacts (ECM). This morphological typing of the contact sites allowed us to recognize and assign distinctive immunolabeling patterns for the three proteins to each type of site on the frozen sections. (b) FA sites were immunolabeled intracellularly for vinculin and alpha-actinin, with vinculin labeling situated closer to the membrane than alpha-actinin. Fibronectin was not labeled in the narrow gap between the cell surface and the substratum, or between two cells, at FA sites. Control experiments showed that this could not be ascribed to inaccessibility of the FA narrow gap to the immunolabeling reagents but indicated an absence or severe depletion of fibronectin from these sites. (c) CC sites were labeled intracellularly for alpha-actinin but not vinculin and were labeled extracellularly for fibronectin. (d) ECM sites were characterized by large separations (often greater than 100 nm) between the cell and substratum or between two cells, which were connected by long cables of extracellular matrix components, including fibronectin. In late (24-36 h) cultures, ECM contacts predominated over the other types. ECM sites appeared to be of two kinds, one labeled intracellularly for both alpha-actinin and vinculin, the other for alpha-actinin alone. (e) From these and other results, a coherent but tentative scheme is proposed for the molecular ultrastructure of these contacts sites, and specific functional roles are suggested for fibronectin, vinculin, and alpha-actinin in cell adhesion and in the linkage of intracellular microfilaments to membranes at the different types of contact sites.


An ergonomic evaluation comparing desktop, notebook, and subnotebook computers.
To evaluate and compare the postures and movements of the cervical and upper thoracic spine, the typing performance, and workstation ergonomic factors when using a desktop, notebook, and subnotebook computers. Repeated-measures design. A motion analysis laboratory with an electromagnetic tracking device. A convenience sample of 21 university students between ages 20 and 24 years with no history of neck or shoulder discomfort. Each subject performed a standardized typing task by using each of the 3 computers. Measurements during the typing task were taken at set intervals. Cervical and thoracic spines adopted a more flexed posture in using the smaller-sized computers. There were significantly greater neck movements in using desktop computers when compared with the notebook and subnotebook computers. The viewing distances adopted by the subjects decreased as the computer size decreased. Typing performance and subjective rating of difficulty in using the keyboards were also significantly different among the 3 types of computers. Computer users need to consider the posture of the spine and potential risk of developing musculoskeletal discomfort in choosing computers.


In Vitro Cultivation of 'Unculturable' Oral Bacteria, Facilitated by Community Culture and Media Supplementation with Siderophores.
Over a third of oral bacteria are as-yet-uncultivated in-vitro. Siderophores have been previously shown to enable in-vitro growth of previously uncultivated bacteria. The objective of this study was to cultivate novel oral bacteria in siderophore-supplemented culture media. Various compounds with siderophore activity, including pyoverdines-Fe-complex, desferricoprogen and salicylic acid, were found to stimulate the growth of difficult-to-culture strains Prevotella sp. HOT-376 and Fretibacterium fastidiosum. Furthermore, pyrosequencing analysis demonstrated increased proportions of the as-yet-uncultivated phylotypes Dialister sp. HOT-119 and Megasphaera sp. HOT-123 on mixed culture plates supplemented with siderophores. Therefore a culture model was developed, which incorporated 15 μg siderophore (pyoverdines-Fe-complex or desferricoprogen) or 150 μl neat subgingival-plaque suspension into a central well on agar plates that were inoculated with heavily-diluted subgingival-plaque samples from subjects with periodontitis. Colonies showing satellitism were passaged onto fresh plates in co-culture with selected helper strains. Five novel strains, representatives of three previously-uncultivated taxa (Anaerolineae bacterium HOT-439, the first oral taxon from the Chloroflexi phylum to have been cultivated; Bacteroidetes bacterium HOT-365; and Peptostreptococcaceae bacterium HOT-091) were successfully isolated. All novel isolates required helper strains for growth, implying dependence on a biofilm lifestyle. Their characterisation will further our understanding of the human oral microbiome.


Multiple epiphyseal dysplasia.
Multiple epiphyseal dysplasia is a genotypically and phenotypically heterogeneous disorder affecting the epiphysis of long bones. Inheritance may be autosomal dominant or autosomal recessive. Autosomal dominant variants include mutations of the collagen oligomeric matrix protein, collagen type IX α-1, collagen type IX α-2, collagen type IX α-3, and matrilin-3 genes. The autosomal recessive variant is caused by a mutation of the sulfate transporter gene SLC26A2. These mutations cause disorganized endochondral ossification of the epiphysis, ultimately leading to destruction of the articular cartilage. Patients typically present in childhood, but some may not present until early adulthood. A presumptive diagnosis can be made with clinical history, physical examination, detailed family history, and radiographic findings. Definitive diagnosis requires genetic testing. Treatment is based on the age of the patient, the amount of limb deformity, the level of joint destruction, and the needs of the patient. Children can greatly benefit from limb realignment procedures, and adults can have excellent outcomes with joint arthroplasty.


Carrier density and Schottky barrier on the performance of DC nanogenerator.
By assembling a ZnO nanowire (NW) array based nanogenerator (NG) that is transparent to UV light, we have investigated the performance of the NG by tuning its carrier density and the characteristics of the Schottky barrier at the interface between the metal electrode and the NW. The formation of a Schottky diode at the interface is a must for the effective operation of the NG. UV light not only increases the carrier density in ZnO but also reduces the barrier height. A reduced barrier height greatly weakens the function of the barrier for preserving the piezoelectric potential in the NW for an extended period of time, resulting in little output current. An increased carrier density speeds up the rate at which the piezoelectric charges are screened/neutralized, but a very low carrier density prevents the flow of current through the NWs. Therefore, there is an optimum conductance of the NW for maximizing the output of the NG. Our study provides solid evidence to further prove the mechanism proposed for the piezoelectric NG and piezotronics. The output current density of the NG has been improved to 8.3 microA/cm2.


Osmoregulation of vasopressin secretion is altered in the postacute phase of septic shock.
To determine whether septic shock patients have an abnormal reponse to increasing osmolarity. Prospective interventional study. Intensive care unit at Raymond Poincaré and Etampes Hospitals. Normonatremic patients at > 72 hrs from septic shock onset. Osmotic challenge consisting of infusing 500 mL of hypertonic saline solution (with cumulative amount of sodium not exceeding 24 g) over 120 mins. Plasma arginine vasopressin levels were measured 15 mins before the test and then four times every 30 mins. A slope of the relation between arginine vasopressin and plasma sodium levels of < 0.5 pg/mEq defined nonresponders. Among the 33 included patients, 17 (52%) were nonresponders. During osmotic challenge, variations throughout the test in plasma sodium levels, blood pressure, and central venous pressure were comparable between the two groups. Arginine vasopressin increased from 4.8 pg/mL [3.3-6.4 pg/mL] to 14.4 pg/mL [11.2-23.3 pg/mL] in responders but only from 2.8 pg/mL [2.3-4.0 pg/mL] to 4.0 pg/mL [3.1-5.3 pg/mL] in nonresponders (p < .0001). Responders had a higher plasma arginine vasopressin levels at baseline and a more severe hematosis alteration. Nonresponders had more frequently bacteremia and liver dysfunction, been referred from the ward and undergone surgery. Critical illness severity, hemodynamic alteration, hydroelectrolytic disturbances, treatment, and outcome did not differ between the two groups. Osmoregulation is dramatically altered in half of patients with prolonged septic shock.


Epizootic cutaneous papillomatosis in roach Rutilus rutilus: sex and size dependence, seasonal occurrence and between-population differences.
Epidemiology of epidermal papillomatosis in roach Rutilus rutilus was studied in 11 roach populations in Finland in 1999 and 2000 during the spawning period. In addition, the seasonal pattern of the disease was described in 1 population. Papilloma tumours were observed on the skin and fins of roach in 8 populations out of 11. Prevalence of papillomatosis varied from 0 to 62%, being higher in populations subject to industrial or sewage effluents. Males and large fish had higher prevalence of papilloma tumours. The disease outbreak peaked during the spawning period and signs of recovered skin on fish were observed later during the year. The gender dependence in the disease prevalence found in the study may indicate differences in stress and immunocompetence between roach males and females during the spawning period.


Are accuracy and reaction time affected via different processes?
A recent study by van Ede et al. (2012) shows that the accuracy and reaction time in humans of tactile perceptual decisions are affected by an attentional cue via distinct cognitive and neural processes. These results are controversial as they undermine the notion that accuracy and reaction time are influenced by the same latent process that underlie the decision process. Typically, accumulation-to-bound models (like the drift diffusion model) can explain variability in both accuracy and reaction time by a change of a single parameter. To elaborate the findings of van Ede et al., we fitted the drift diffusion model to their behavioral data. Results show that both changes in accuracy and reaction time can be partly explained by an increase in the accumulation of sensory evidence (drift rate). In addition, a change in non-decision time is necessary to account for reaction time changes as well. These results provide a subtle explanation of how the underlying dynamics of the decision process might give rise to differences in both the speed and accuracy of perceptual tactile decisions. Furthermore, our analyses highlight the importance of applying a model-based approach, as the observed changes in the model parameters might be ecologically more valid, since they have an intuitive relationship with the neuronal processes underlying perceptual decision making.


Pregnancy related complications in women with hypertrophic cardiomyopathy.
To determine whether pregnancy is well tolerated in hypertrophic cardiomyopathy. Referral clinic. The study cohort comprised 127 consecutively referred women with hypertrophic cardiomyopathy. Forty (31.5%) underwent clinical evaluation before pregnancy. The remaining 87 (68.5%) were referred after their first pregnancy. All underwent history, examination, electrocardiography, and echocardiography. Pregnancy related symptoms and complications were determined by questionnaire and review of medical and obstetric records where available. There were 271 pregnancies in total. Thirty six (28.3%) women reported cardiac symptoms in pregnancy. Over 90% of these women had been symptomatic before pregnancy. Symptoms deteriorated during pregnancy in fewer than 10%. Of the 36 women with symptoms during pregnancy, 30 had further pregnancies. Symptoms reoccurred in 18 (60%); symptomatic deterioration was not reported. Heart failure occurred postnatally in two women (1.6%). No complications were reported in 19 (15%) women who underwent general anaesthesia and in 22 (17.4%) women who received epidural anaesthesia, three of whom had a significant left ventricular outflow tract gradient at diagnosis after pregnancy. Three unexplained intrauterine deaths occurred in women taking cardiac medication throughout pregnancy. No echocardiographic or clinical feature was a useful indicator of pregnancy related complications. Most women with hypertrophic cardiomyopathy tolerate pregnancy well. However, rare complications can occur and therefore planned delivery and fetal monitoring are still required for some patients.


Artificial microRNA-mediated resistance against the monopartite begomovirus Cotton leaf curl Burewala virus.
Cotton leaf curl disease, caused by single-stranded DNA viruses of the genus Begomovirus (family Geminiviridae), is a major constraint to cotton cultivation across Pakistan and north-western India. At this time only cotton varieties with moderate tolerance are available to counter the disease. microRNAs (miRNAs) are a class of endogenous small RNA molecules that play an important role in plant development, signal transduction, and response to biotic and a biotic stress. Studies have shown that miRNAs can be engineered to alter their target specificity. Such artificial miRNAs (amiRNAs) have been shown to provide resistance against plant-infecting viruses. Two amiRNA constructs, based on the sequence of cotton miRNA169a, were produced containing 21 nt of the V2 gene sequence of Cotton leaf curl Burewala virus (CLCuBuV) and transformed into Nicotiana benthamiana. The first amiRNA construct (P1C) maintained the miR169a sequence with the exception of the replaced 21 nt whereas in the second (P1D) the sequence of the miRNA169a backbone was altered to restore some of the hydrogen bonding of the mature miRNA duplex. P1C transgenic plants showed good resistance when challenge with CLCuBV; plants being asymptomatic with low viral DNA levels. The resistance to heterologous viruses was lower and correlated with the numbers of sequence mismatches between the amiRNA and the V2 gene sequence. P1D plants showed overall poorer resistance to challenge with all viruses tested. The results show that the amiRNA approach can deliver efficient resistance in plants against a monopartite begomoviruses and that this has the potential to be broad-spectrum, providing protection from a number of viruses. Additionally the findings indicate that the levels of resistance depend upon the levels of complementarity between the amiRNA and the target sequence and the sequence of the miRNA backbone, consistent with earlier studies.


MLC-inhibiting antibodies in mice after blood transfusions.
Balb/c (H-2d) mice were transfused weekly with 0.1 ml of whole blood from C3H/HeJ (C3H) (H-2k) mice. One and two weeks after each transfusion, mice were bled and the sera were collected and pooled. Serum samples from transfused mice were absorbed with erythrocytes and spleen cells from C3H mice, and then heat-inactivated. The presence of antiidiotypic antibodies in these sera was investigated using the mixed lymphocyte culture (MLC) inhibition test, in which spleen cells from normal Balb/c mice were tested for proliferative responses to x-irradiated C3H stimulator cells in the presence of sera from transfused mice. Sera obtained from transfused mice caused significant inhibition of responses in MLC. This inhibition in MLC was specific for stimulator cells from the blood donor (C3H), and little or no inhibition was observed with stimulator cells from third-party SJL mice. In addition, the inhibitory effect in MLC was specific for responder cells from the recipient Balb/c mice and no inhibition was observed with responder cells from blood donor C3H mice. These results suggest that blood transfusions induce antiidiotypic antibodies that can block the T cell antigen-specific receptors and cause inhibition of responses of the recipient mice to blood donor alloantigens in MLC. Thus in this strain combination, disparate for the entire H-2 region, antiidiotypic antibodies developed after 1 blood transfusion. These findings are in contrast to our earlier published results in which antiidiotypic antibodies developed after 3 transfusions in recipient-blood donor combination incompatible for the K and I regions. These data suggest that the development of antiidiotypic antibodies may be related to the level of histoincompatibility at the H-2 complex between recipient and the blood donor.


Synthesis and evaluation of fluorescent heterocyclic aminoadamantanes as multifunctional neuroprotective agents.
A series of fluorescent heterocyclic adamantane amines were synthesised with the goal to develop novel fluorescent ligands for neurological assay development. These derivatives demonstrated multifunctional neuroprotective activity through inhibition of the N-methyl-d-aspartate receptor/ion channel, calcium channels and the enzyme nitric oxide synthase. It also exhibited a high degree of free radical scavenging potential. N-(1-adamantyl)-2-oxo-chromene-3-carboxamide (8), N-adamantan-1-yl-5-dimethyl-amino-1-naphthalenesulfonic acid (11) and N-(1-cyano-2H-isoindol-2-yl) adamantan-1-amine (12) were found to possess a high degree of multifunctionality with favourable physical-chemical properties for bioavailability and blood-brain barrier permeability. The ability of these heterocyclic adamantane amine derivatives as nitric oxide synthase inhibitors, calcium channel modulators, NMDAR inhibitors and effective antioxidants, indicate that they may find application as multifunctional drugs in neuroprotection.


Interface phonon modes in the [AlN/GaN]20 and [Al0.35Ga0.65N/Al0.55Ga0.45N]20 2D multi-quantum well structures.
The observation of interface (IF) phonon modes in the recorded Raman spectra of c-plane oriented [AlN/GaN]20 and [Al0.35Ga0.65N/Al0.55Ga0.45N]20 multi-quantum well (MQW) structures grown via plasma assisted molecular beam epitaxy is reported. The nominal shift in the IF phonon mode of E1 symmetry for [Al0.35Ga0.65N/Al0.55Ga0.45N]20 compared to that of the [AlN/GaN]20 MQW structure is understood on the basis of change in dielectric constants (εm) of the surrounding medium. The presence of buffer layers in [Al0.35Ga0.65N/Al0.55Ga0.45N]20 MQW over a sapphire substrate is also understood by characterizing the IF phonon mode of A1 symmetry. The observed IF phonon modes in the spectra are attributed to the relaxation of Raman selection rules away from the Brillouin zone centre because of the breakdown of the translational symmetry of surface potential due to the presence of the periodic interfaces and surface modulations in the superlattice structures of MQWs. The corresponding required edge lengths (L) of 2D plates, for the observation of the breakdown of the surface potential, are computed from the simulated dispersion relation curve of IF modes. The integral multiples of uniformly distributed platelets, originated due to the horizontal uneven irregularities on the surface of superlattices, are matched with the calculated L values.


Castration fever: On trans, China, and psychoanalysis.
This essay considers the evolution of the author's research over the last 15 years in which the treatment of castration as a historical problem holds promise for bridging disparate scholarly fields and paradigms. In particular, by tracing the shift in the author's intellectual focus from the science of sex change to the history of transcultural psychoanalysis, some insights are offered in regard to the intertwined politics of transness, Chineseness, and the unconscious. Though psychoanalysis may appear as a subject far removed from the eunuchs of ancient China, this essay highlights some of the methodological stakes that have saturated the historical study of both topics. These reflections can serve as a touchstone for thinking beyond disciplinary norms and conventions, especially in Chinese studies and the history of science.


Seasonal trends in antidepressant prescribing, depression, anxiety and self-harm in adolescents and young adults: an open cohort study using English primary care data.
There is an increasing demand for mental health services for young people, which may vary across the year. To determine whether there are seasonal patterns in primary care antidepressant prescribing and mental health issues in adolescents and young adults. This cohort study used anonymised electronic health records from general practices in England contributing to QResearch. It included 5 081 263 males and females aged 14-18 (adolescents), 19-23 and 24-28 years between 2006 and 2019. The incidence rates per 1000 person-years and the incidence rate ratios (IRRs) were calculated for the first records of a selective serotonin reuptake inhibitor (SSRI) prescription, depression, anxiety and self-harm. The IRRs were adjusted for year, region, deprivation, ethnic group and number of working days. There was an increase in SSRI prescribing, depression and anxiety incidence in male and female adolescents in the autumn months (September-November) that was not seen in older age groups. The IRRs for SSRI prescribing for adolescents peaked in November (females: 1.75, 95% CI 1.67 to 1.83, p<0.001; males: 1.72, 95% CI 1.61 to 1.84, p<0.001, vs in January) and for depression (females: 1.29, 95% CI 1.25 to 1.33, p<0.001; males: 1.29, 95% CI 1.23 to 1.35, p<0.001). Anxiety peaked in November for females aged 14-18 years (1.17, 95% CI 1.13 to 1.22, p<0.001) and in September for males (1.19, 95% CI 1.12 to 1.27, p<0.001). There were higher rates of antidepressant prescribing and consultations for depression and anxiety at the start of the school year among adolescents. Support around mental health issues from general practitioners and others should be focused during autumn.


Evidence for interplay between genes and maternal stress in utero: monoamine oxidase A polymorphism moderates effects of life events during pregnancy on infant negative emotionality at 5 weeks.
The low activity variant of the monoamine oxidase A (MAOA) functional promoter polymorphism, MAOA-LPR, in interaction with adverse environments (G × E) is associated with child and adult antisocial behaviour disorders. MAOA is expressed during foetal development so in utero G × E may influence early neurodevelopment. We tested the hypothesis that MAOA G × E during pregnancy predicts infant negative emotionality soon after birth. In an epidemiological longitudinal study starting in pregnancy, using a two stage stratified design, we ascertained MAOA-LPR status (low vs. high activity variants) from the saliva of 209 infants (104 boys and 105 girls), and examined predictions to observed infant negative emotionality at 5 weeks post-partum from life events during pregnancy. In analyses weighted to provide estimates for the general population, and including possible confounders for life events, there was an MAOA status by life events interaction (P = 0.017). There was also an interaction between MAOA status and neighbourhood deprivation (P = 0.028). Both interactions arose from a greater effect of increasing life events on negative emotionality in the MAOA-LPR low activity, compared with MAOA-LPR high activity infants. The study provides the first evidence of moderation by MAOA-LPR of the effect of the social environment in pregnancy on negative emotionality in infancy, an early risk for the development of child and adult antisocial behaviour disorders.


Orai1 and Orai3 in Combination with Stim1 Mediate the Majority of Store-operated Calcium Entry in Astrocytes.
Astrocytes are non-excitable cells in the brain and their activity largely depends on the intracellular calcium (Ca2+) level. Therefore, maintaining the intracellular Ca2+ homeostasis is critical for proper functioning of astrocytes. One of the key regulatory mechanisms of Ca2+ homeostasis in astrocytes is the store-operated Ca2+ entry (SOCE). This process is mediated by a combination of the Ca2+-store-depletion-sensor, Stim, and the store-operated Ca2+-channels, Orai and TrpC families. Despite the existence of all those families in astrocytes, previous studies have provided conflicting results on the molecular identification of astrocytic SOCE. Here, using the shRNA-based gene-silencing approach and Ca2+-imaging from cultured mouse astrocytes, we report that Stim1 in combination with Orai1 and Orai3 contribute to the major portion of astrocytic SOCE. Gene-silencing of Stim1 showed a 79.2% reduction of SOCE, indicating that Stim1 is the major Ca2+-store-depletion-sensor. Further gene-silencing showed that Orai1, Orai2, Orai3, and TrpC1 contribute to SOCE by 35.7%, 20.3%, 26.8% and 12.2%, respectively. Simultaneous gene-silencing of all three Orai subtypes exhibited a 67.6% reduction of SOCE. Based on the detailed population analysis, we predict that Orai1 and Orai3 are expressed in astrocytes with a large SOCE, whereas TrpC1 is exclusively expressed in astrocytes with a small SOCE. This analytical approach allows us to identify the store operated channel (SOC) subtype in each cell by the degree of SOCE. Our results propose that Stim1 in combination with Orai1 and Orai3 are the major molecular components of astrocytic SOCE under various physiological and pathological conditions.


Nationwide surveillance of antimicrobial susceptibility patterns of pathogens isolated from surgical site infections (SSI) in Japan.
To investigate the trends of antimicrobial resistance in pathogens isolated from surgical site infections (SSI), a Japanese surveillance committee conducted the first nationwide survey. Seven main organisms were collected from SSI at 27 medical centers in 2010 and were shipped to a central laboratory for antimicrobial susceptibility testing. A total of 702 isolates from 586 patients with SSI were included. Staphylococcus aureus (20.4 %) and Enterococcus faecalis (19.5 %) were the most common isolates, followed by Pseudomonas aeruginosa (15.4 %) and Bacteroides fragilis group (15.4 %). Methicillin-resistant S. aureus among S. aureus was 72.0 %. Vancomycin MIC 2 μg/ml strains accounted for 9.7 %. In Escherichia coli, 11 of 95 strains produced extended-spectrum β-lactamase (Klebsiella pneumoniae, 0/53 strains). Of E. coli strains, 8.4 % were resistant to ceftazidime (CAZ) and 26.3 % to ciprofloxacin (CPFX). No P. aeruginosa strains produced metallo-β-lactamase. In P. aeruginosa, the resistance rates were 7.4 % to tazobactam/piperacillin (TAZ/PIPC), 10.2 % to imipenem (IPM), 2.8 % to meropenem, cefepime, and CPFX, and 0 % to gentamicin. In the B. fragilis group, the rates were 28.6 % to clindamycin, 5.7 % to cefmetazole, 2.9 % to TAZ/PIPC and IPM, and 0 % to metronidazole (Bacteroides thetaiotaomicron; 59.1, 36.4, 0, 0, 0 %). MIC₉₀ of P. aeruginosa isolated 15 days or later after surgery rose in TAZ/PIPC, CAZ, IPM, and CPFX. In patients with American Society of Anesthesiologists (ASA) score ≥3, the resistance rates of P. aeruginosa to TAZ/PIPC and CAZ were higher than in patients with ASA ≤2. The data obtained in this study revealed the trend of the spread of resistance among common species that cause SSI. Timing of isolation from surgery and the patient's physical status affected the selection of resistant organisms.


Proteins, Transcripts, and Genetic Architecture of Seminal Fluid and Sperm in the Mosquito Aedes aegypti.
The yellow fever mosquito, Aedes aegypti,, transmits several viruses causative of serious diseases, including dengue, Zika, and chikungunya. Some proposed efforts to control this vector involve manipulating reproduction to suppress wild populations or to replace them with disease-resistant mosquitoes. The design of such strategies requires an intimate knowledge of reproductive processes, yet our basic understanding of reproductive genetics in this vector remains largely incomplete. To accelerate future investigations, we have comprehensively catalogued sperm and seminal fluid proteins (SFPs) transferred to females in the ejaculate using tandem mass spectrometry. By excluding female-derived proteins using an isotopic labeling approach, we identified 870 sperm proteins and 280 SFPs. Functional composition analysis revealed parallels with known aspects of sperm biology and SFP function in other insects. To corroborate our proteome characterization, we also generated transcriptomes for testes and the male accessory glands-the primary contributors to Ae. aegypti, sperm and seminal fluid, respectively. Differential gene expression of accessory glands from virgin and mated males suggests that transcripts encoding proteins involved in protein translation are upregulated post-mating. Several SFP transcripts were also modulated after mating, but >90% remained unchanged. Finally, a significant enrichment of SFPs was observed on chromosome 1, which harbors the male sex determining locus in this species. Our study provides a comprehensive proteomic and transcriptomic characterization of ejaculate production and composition and thus provides a foundation for future investigations of Ae. aegypti, reproductive biology, from functional analysis of individual proteins to broader examination of reproductive processes.


Neural control of maternal and paternal behaviors.
Parental care, including feeding and protection of young, is essential for the survival as well as mental and physical well-being of the offspring. A large variety of parental behaviors has been described across species and sexes, raising fascinating questions about how animals identify the young and how brain circuits drive and modulate parental displays in males and females. Recent studies have begun to uncover a striking antagonistic interplay between brain systems underlying parental care and infant-directed aggression in both males and females, as well as a large range of intrinsic and environmentally driven neural modulation and plasticity. Improved understanding of the neural control of parental interactions in animals should provide novel insights into the complex issue of human parental care in both health and disease.


Electro-osmotic flow over a charged superhydrophobic surface.
Bubbles can be trapped inside textured structures such as grooves, forming a superhydrophobic surface. A superhydrophobic surface has a large effective hydrodynamic slip length compared to a smooth hydrophobic surface and holds the promise of enhancing electrokinetic flows that find many interesting applications in microfluidics. However, recent theoretical studies suggested that electro-osmotic flows over a weakly charged superhydrophobic surface [the zeta potential of the surface is smaller than the thermal potential (25 mV)] can only be enhanced when liquid-gas interfaces are charged [T. M. Squires, Phys. Fluids 20, 092105 (2008); Bahga, J. Fluid Mech. 644, 245 (2010)]. So far there is little work reported when the zeta potential of the surface is comparable or even larger than the thermal potential. In this paper we numerically investigate electro-osmotic flows over a periodically striped slip-stick surface by solving the standard Poisson-Nernst-Planck equations. Our results indicate that at large zeta potentials, even if liquid-gas interfaces are charged, the nonuniform surface conduction due to the mismatch between surface conductions over no-shear and no-slip regions leads to electric field lines penetrating the double layer and thus the nonuniform surface conduction weakens the tangential component of the electric field which primarily drives electro-osmotic flows. Our results imply that, in the presence of strong nonuniform surface conduction, enhanced electro-osmotic flows over a superhydrophobic surface are possible only in certain conditions. In particular, the enhancement due to the slip can potentially be lost at large zeta potentials. Similar loss of the enhancement of a charged particle's electrophoretic mobility due to the slip was reported by Khair and Squires [Phys. Fluids 21, 042001 (2009)].


Paramphistomum daubneyi: characteristics of infection in three lymnaeid species.
Experimental infections of two South American lymnaeids (Lymnaea neotropica and L. viatrix var. ventricosa) with Paramphistomum daubneyi were carried out to determine if these snail species could sustain larval development of this digenean and, if so, to specify their potential for cercarial production. A French population of Galba truncatula infected and raised according to the same protocol served as controls. In both experiments, prevalence of P. daubneyi infections in snails did not significantly differ from each other. In snail groups evaluated for cercarial shedding (first experiment), a significantly lower number of shed cercariae was noted for L. neotropica, while those from G. truncatula and L. v. ventricosa did not differ significantly from each other. Dissection of infected snails at day 65 post-exposure at 20°C (second experiment) found significantly lower burdens of P. daubneyi rediae and cercariae in the bodies of L. neotropica than in those of G. truncatula and L. v. ventricosa. Compared to total cercarial production observed in dissected snails, the percentage of cercariae which exited from snails was 75.6% for G. truncatula, 21.6% for L. neotropica, and 91.4% for L. v. ventricosa. This last species seems to be a good candidate for metacercarial production of P. daubneyi.


The emerging utility of animal models of chronic neurodegenerative diseases.
The two most common neurodegenerative diseases are Alzheimer's disease (AD) and Parkinson's disease (PD). The symptoms are caused by the initially selective degeneration of neuronal subpopulations involved in memory (AD) or movement control (PD). The cause of both diseases is unknown, but ageing is an inevitable risk factor. The identification of disease-associated genes was a breakthrough for the understanding of molecular mechanisms of neurodegeneration and has provided the basis for the establishment of cell culture and animal model systems, instrumental for target validation and drug screening. Familial AD is caused by mutations in the beta-amyloid precursor protein (betaAPP) and in the gene products responsible for its proteolytic processing, namely the presenilins. Transgenic mice expressing these mutant genes develop characteristic AD plaques in an age-dependent manner. A reduction of plaque burden and amelioration of cognitive decline in these animals was recently achieved by vaccination with amyloid beta-protein fibrils. The other hallmark lesion of AD, the neurofibrillary tangle, has been modelled recently in transgenic mice expressing mutant tau protein linked to frontotemporal dementia. PD is characterised by intraneuronal cytoplasmic deposits (Lewy bodies) of the PD-associated gene product alpha-synuclein. Transgenic expression of alpha-synuclein recreated hallmark features of PD in mice and fruit flies, establishing alpha-synuclein as PD-causing drug target. Moreover, environmental risk factors such as the pesticide rotenone have been used successfully to generate rodent models of PD. Lesion models of PD are being exploited for the development of experimental gene therapy and transplantation approaches.


Susceptibility of various Japanese freshwater fish species to an isolate of viral haemorrhagic septicaemia virus (VHSV) genotype IVb.
Genotype IVb of viral haemorrhagic septicaemia virus (VHSV) was isolated for the first time in the Great Lakes basin in 2003, where it spread and caused mass mortalities in several wild fish species throughout the basin. In order to prevent further spreading of the disease and to assess risks of new genotypes invading new watersheds, basic microbiological information such as pathogenicity studies are essential. In this study, experimental infections were conducted on 7 indigenous freshwater fish species from Japan by immersion with a VHSV genotype IVb isolate. In Expt 1, cumulative mortalities in bluegill Lepomis macrochirus used as positive controls, Japanese fluvial sculpin Cottus pollux, and iwana Salvelinus leucomaenis pluvius were 50, 80 and 0%, respectively. In Expt 2, cumulative mortalities of 100, 100 and 10% were observed in Japanese fluvial sculpin C. pollux, Japanese rice fish Oryzias latipes and yoshinobori Rhinogobius sp., respectively. No mortality was observed in honmoroko Gnathopogon caerulescens, akaza Liobagrus reini or Japanese striped loach Cobitis biwae. VHSV was detected by RT-PCR from samples of kidney, spleen, and brain from all dead fish, and virus re-isolation by cell culture was successful from all dead fish. We detected the virus in the brain from a few surviving bluegill 50 d post exposure by both cell culture and RT-PCR. These results revealed that VHSV IVb could become a serious threat to wild freshwater fish species in Japan, and that some surviving fish might become healthy carriers of the virus.


Serum adiponectin concentrations predict the developments of type 2 diabetes and the metabolic syndrome in elderly Koreans.
To determine whether low serum adiponectin concentrations are able to predict the future developments of type 2 diabetes and the metabolic syndrome using the National Cholesterol Education Programme (NCEP) criteria. We also investigated the stability of adiponectin levels and the relationships between baseline adiponectin levels and changes in the parameters related to the metabolic syndrome over a period of 3 years. This prospective cohort study included 372 elderly Koreans who participate in the SWS (South-West Seoul) study, which was conducted in 1999 and 2002 in Seoul, Korea. Fasting and postchallenge 2-h plasma glucose, body mass index (BMI), waist-to-hip ratio (WHR), blood pressure, lipid profiles and serum adiponectin data obtained in 1999 and 2002 were examined. The within-person variation between 1999 and 2002 of serum adiponectin was not significant (P = 0.61). Serum adiponectin was closely correlated with the risks factors of cardiovascular disease (CVD), that is negatively with BMI, WHR, blood pressure, triglyceride and blood glucose levels, and positively with high density lipoprotein (HDL)-cholesterol levels. Subjects with the metabolic syndrome showed lower serum adiponectin concentrations than those without the metabolic syndrome (P < 0.0001). Baseline adiponectin levels were found to be correlated with subsequent changes in WHR, low density lipoprotein (LDL)-cholesterol, fasting and postload 2-h glucose over the 3-year period, after adjusting for baseline values. Multiple logistic regression analysis showed that lower baseline serum adiponectin concentrations were significantly associated with the developments of type 2 diabetes and the metabolic syndrome after adjusting for age, sex, obesity, history of impaired fasting glucose (IFG) or impaired glucose tolerance (IGT), hypertension and dyslipidaemia. Reduced concentrations of adiponectin were found to be independently associated with increased risk of both type 2 diabetes and the metabolic syndrome in elderly Koreans.


Preliminary results of the solar flux radiometer experiment aboard the pioneer venus multiprobe mission.
The solar flux radiometer aboard the Pioneer Venus large probe operated successfully during its descent through the atmosphere of Venus. Upward, downward, and net fluxes from 0.4 to 1.0 micrometers were obtained at more than 390 levels between 185 millibars (at an altitude of approximately 61 kilometers) and the surface. Fluxes from 0.4 to 1.8 micrometers were also obtained between 185 millibars and about the level at which the pressure was 2 atmospheres. Data from 80 to 185 millibars should be available after additional decoding by the Deep Space Network. Upward and downward intensities in a narrower band from 0.59 to 0.66 micrometers were also obtained throughout the descent in order to constrain cloud properties. The measurements indicate three cloud regions above the 1.3-atmosphere level (at an altitude of approximately 49 kilometers) and a clear atmosphere beneath that level. At the 67 degrees solar zenith of the probe entry site, some 15 watts per square meter are absorbed at the surface by a dark ground, which implies that about 2 percent of the solar energy incident on the planet is absorbed at the ground.


The influence of diet on faecal DNA amplification and sex identification in brown bears (Ursus arctos).
To evaluate the influence of diet on faecal DNA amplification, 11 captive brown bears (Ursus arctos) were placed on six restricted diets: grass (Trifolium spp., Haplopappus hirtus and Poa pratensis), alfalfa (Lupinus spp.), carrots (Daucus spp.), white-tailed deer (Odocoileus virginianus), blueberries (Vaccinium spp.) and salmon (Salmo spp.). DNA was extracted from 50 faecal samples of each restricted diet, and amplification of brown bear DNA was attempted for a mitochondrial DNA (mtDNA) locus and nuclear DNA (nDNA) locus. For mtDNA, no significant differences were observed in amplification success rates across diets. For nDNA, amplification success rates for salmon diet extracts were significantly lower than all other diet extracts (P < 0.001). To evaluate the accuracy of faecal DNA sex identification when female carnivores consume male mammalian prey, female bears were fed male white-tailed deer. Four of 10 extracts amplified, and all extracts were incorrectly scored as male due to amplification of X and Y-chromosome fragments. The potential biases highlighted in this study have broad implications for researchers using faecal DNA for individual and sex identification, and should be evaluated in other species.


Nerve growth factor increases electrical activity of neural cells derived from murine bone marrow stromal cells.
Nerve growth factor (NGF) triggers long-term neuronal excitability. We examined its effect on murine bone marrow stromal cells (BMSC)-derived neurons. With an optimal differentiation protocol, BMSCs were differentiated into neurons in culture. To confirm the probability of differentiation of BMSC into neuron, the expression of neuronal marker protein, neurofilament, was examined by immunocytochemistry. To examine the electrophysiological properties of BMSC-derived neurons, the field potentials were recorded either from nontreated (control) BMSC-derived neurons or from BMSC-derived neurons after the treatment with NGF by using extracellular recording techniques. Most BMSC-derived neurons showed spontaneous discharges whose amplitudes were up to 2 mV. When NGF at a concentration of 100 ng/ml was applied to BMSC-derived neurons, the amplitudes of discrete field potentials were gradually enlarged within 1 min after NGF application and peaked 3 min later (20-fold the size of control). However, the enlargement of the amplitudes of field potentials almost disappeared 5 min after NGF application. This finding indicates that neuronal cells derived from murine BMSCs generate discrete field potential activities spontaneously and that NGF has the effect of enlarging transient, but not sustained, electrical activity of BMSC-derived neurons.


Super CitriMax (HCA-SX) attenuates increases in oxidative stress, inflammation, insulin resistance, and body weight in developing obese Zucker rats.
Super CitriMax (HCA-SX) is a novel calcium/potassium salt of (-)-hydroxycitric acid extracted from the dried fruit rind of the plant Garcinia cambogia, and commonly consumed as weight loss dietary supplement. In the present study, we investigated the effect of HCA-SX on inflammation, oxidative stress and insulin resistance in developing obese Zucker rats, an animal model of type II diabetes associated with inflammation and oxidative stress. Male Zucker rats (5-week old) were supplemented with vehicle (control) and HCA-SX in drinking water for 7 weeks. Oxidative stress markers, including malondialdehyde (MDA), protein carbonyl (DNPH), and protein tyrosine nitration (tyr-NO(2)) were measured in the liver and kidney tissues using biochemical and immunoblotting techniques. Compared to controls, the levels of MDA, DNPH and tyr-NO(2) were lower in the liver and kidney of HCA-SX-treated animals. Furthermore, the levels of C-reactive protein and interleukin-6, markers of inflammation measured by ELISA, were lower in the plasma of HCA-SX-supplemented animals compared to controls, as were levels of fasting plasma insulin, glucose, and triglycerides. Interestingly, insulin resistance did not develop in HCA-SX-supplemented rats. Food-intake and body weight gain was also lower in rats supplemented with HCA-SX compared to their control counterparts. These results suggest that HCA-SX supplementation in obese Zucker rats reduces food-intake, body weight gain, and also attenuates the increases in inflammation, oxidative stress, and insulin resistance observed in untreated animals. Therefore, HCA-SX may be used as an intervention to overcome obesity-related complications, including inflammation, oxidative stress, and insulin resistance.


Estimating the time-varying generation rate of acetic acid from an all-purpose floor cleaner.
Understanding the relationship between consumer product use and risk of adverse health outcomes facilitates appropriate risk management and product stewardship. A preferred method for estimating the systemic and respiratory tract exposure and dose tailored to cleaning products use has been proposed, refining previously issued exposure guidance. Consistent with other exposure and risk-assessment frameworks, it is dependent upon high-quality exposure determinant data that also serve as model inputs. However, as publicly available exposure determinant data are scarce, the risk assessor is left with the option of estimating determinants such as the generation rate or employing empirical methods to estimate them. When the exposure scenario involves a chemical mixture, estimating the generation rate may not be feasible. We present an approach for estimating the time-varying generation rate of an aqueous acetic acid mixture representative of the base formulation for many consumer and DIY cleaning products that was previously assessed in a screening-level assessment. The approach involved measuring the evaporation rate for a reasonable worst-case scenario under controlled conditions. Knowing the mass applied, a time-varying generation rate was estimated. To evaluate its portability, a field study was conducted in a home where measurements were collected in an all-purpose room with the exterior door open (Room 1) and closed (Room 2), and a bathroom (Room 3) using a portable Fourier Transform Infrared (FTIR) spectrophotometer. Acetic acid concentrations were modeled using two common indoor air models, the Well Mixed Room model. Measured and modeled acetic acid concentrations were compared, with the WMR 95% confidence intervals encompassing measured concentrations for all three rooms, supporting the utility of the approach used and portability of the generation rate derived from it.


In vitro modeling of the blood-brain barrier: simplicity versus complexity.
Proper understanding of blood-brain barrier (BBB) regulation is crucial to reduce/prevent its disruption during injury. Since high brain complexity makes interpretation of in vivo data challenging BBB studies are frequently performed using simplified in vitro models. Although such models represent an important and frequently employed alternative for investigation of BBB function and alterations, our ability to translate in vitro findings to in vivo situation remains sub-optimal. Consequently, despite the fact that our knowledge of the cellular and molecular mechanisms underlying BBB physiology and pathophysiology is constantly increasing, our ability to modulate barrier function remains virtually non-existent. Classical in vitro model systems have provided a wealth of knowledge until now, but it is now evident that newer in vitro models that are more representative of the in vivo situation are needed to further our understanding of barrier physiology. This paper will provide an overview of the BBB cellular components and the most frequently used in vitro BBB model systems. I will discuss their advantages and disadvantages, as well as highlight recently developed models that more closely mimic the BBB in vivo.


Secure attachment to therapist, alliance, and outcome in psychoanalytic psychotherapy with young adults.
Using a novel approach to assess attachment to therapist from patient narratives (Patient Attachment to Therapist Rating Scale; PAT-RS), we investigated the relationships between secure attachment to therapist, patient-rated alliance, and outcome in a sample of 70 young adults treated with psychoanalytic psychotherapy. A series of linear mixed-effects models, controlling for length of therapy and therapist effects, indicated that secure attachment to therapist at termination was associated with improvement in symptoms, global functioning, and interpersonal problems. After controlling for the alliance, these relationships were maintained in terms of symptoms and global functioning. Further, for the follow-up period, we found a suppression effect indicating that secure attachment to therapist predicted continued improvement in global functioning, whereas the alliance predicted deterioration when both variables were modeled together. Although limited by the correlational design, this study suggests that the development of a secure attachment to therapist is associated with treatment gains as well as predictive of posttreatment improvement in functioning. Future research should investigate the temporal development of attachment to therapist and its interaction with alliance and outcome more closely. To ensure differentiation from patient-rated alliance, observer-based measurement of attachment to therapist should be considered.


Analysis of Gga null mice demonstrates a non-redundant role for mammalian GGA2 during development.
Numerous studies using cultured mammalian cells have shown that the three GGAs (Golgi-localized, gamma-ear containing, ADP-ribosylation factor- binding proteins) function in the transport of cargo proteins between the trans- Golgi network and endosomes. However, the in vivo role(s) of these adaptor proteins and their possible functional redundancy has not been analyzed. In this study, the genes encoding GGAs1-3 were disrupted in mice by insertional mutagenesis. Loss of GGA1 or GGA3 alone was well tolerated whereas the absence of GGA2 resulted in embryonic or neonatal lethality, depending on the genetic background of the mice. Thus, GGA2 mediates a vital function that cannot be compensated for by GGA1and/or GGA3. The combined loss of GGA1 and GGA3 also resulted in a high incidence of neonatal mortality but in this case the expression level of GGA2 may be inadequate to compensate for the loss of the other two GGAs. We conclude that the three mammalian GGAs are essential proteins that are not fully redundant.


Recognition of aggressive human behavior using binary local motion descriptors.
Video surveillance is an alternative approach to staff or self-reporting that has the potential to detect and monitor aggressive behaviors more accurately. In this paper, we propose an automatic algorithm capable of recognizing aggressive behaviors from video records using local binary motion descriptors. The proposed algorithm may increase the accuracy for retrieving aggressive behaviors from video records, and thereby facilitates scientific inquiry into this low frequency but high impact phenomenon that eludes other measurement approaches.


A Quantitative Angiographic Comparison of Restenotic Tissue Following Placement of Drug-Eluting Stents and Bare Metal Stents in Symptomatic Patients With Femoropopliteal Disease.
To evaluate the patterns of in-stent restenosis (ISR) within femoropopliteal bare metal stents (BMS) and drug-eluting stents (DES) as determined by quantitative angiographic analysis. Utilizing results from independent core laboratory angiographic imaging analysis, quantitative assessment of the restenotic tissue burden was evaluated in 33 patients with symptomatic femoropopliteal ISR, including 20 lesions in 19 patients (mean age 71.5±8.1 years; 11 men) treated with DES and 14 lesions in 14 patients (mean age 70.6±9.2 years; 9 men) treated with BMS. The average time to target lesion revascularization was similar (8.7 months) for the DES and BMS groups. The DES group had significantly less recurrent disease burden (17.1%) compared with the BMS group (27.8%, p=0.03), representing a 39% relative reduction. Reduced restenotic tissue after endovascular intervention is associated with improved hemodynamics and fewer clinical symptoms and may explain the reduced need for reintervention in restenotic lesions initially treated with DES as compared with BMS. Further study of treatment failure modes may lead to improved device selection criteria to treat patients with peripheral artery disease.


Polymorphisms in chemokine receptor 5 and Toll-like receptor 3 genes are risk factors for clinical tick-borne encephalitis in the Lithuanian population.
Tick-borne encephalitis virus (TBEV) infections can be asymptomatic or cause moderate to severe injuries of the nervous system. We previously reported that a nonfunctional chemokine receptor 5 (CCR5) and a functional Toll-like receptor 3 (TLR3) predispose adults to clinical tick-borne encephalitis (TBE). This study expands our previous findings and further examines polymorphisms in CCR5 and TLR3 genes in different age and disease severity groups. 117 children and 129 adults, stratified into mild, moderate and severe forms of TBE, and 103 adults with severe TBE were analyzed. 135 healthy individuals and 79 patients with aseptic meningoencephalitis served as controls. CCR5 delta 32 and rs3775291 TLR3 genotypes were established by pyrosequencing, and their frequencies were analyzed using recessive genetic, genotype and allelic models. The prevalence of CCR5Δ32 homozygotes was higher in children (2.5%), in adults with severe TBE (1.9%), and in the combined cohort of TBE patients (2.3%) than in controls (0%) (p<0.05). The nonfunctional homozygous TLR3 genotype was less prevalent among the combined TBE cohort (11.5%) than among controls (19.9%) (p = 0.025), but did not differ between children TBE and controls. The genotype and allele prevalence of CCR5 and TLR3 did not differ in children nor adult TBE cohorts stratified by disease severity. However, in the severe adult TBE cohort, homozygous functional TLR3 genotype and wt allele were less prevalent compared to the adult cohort with the whole disease severity spectrum (44.4% vs 59.8% p = 0.022 and 65.2% vs 76.4% p = 0.009; respectively). Independently of age, nonfunctional CCR5Δ32 mutation is a significant risk factor for development of clinical TBE, but not for disease severity. The polymorphism of TLR3 gene predisposes to clinical TBE in adults only and may be associated with disease severity. Further studies are needed to clarify the role of these polymorphisms in susceptibility to TBEV infection.


MRI of the prostate.
Prostate cancer is the most prevalent malignancy in the world. It is the most frequently diagnosed cancer in the American male population and the second most lethal cancer in American men. However, concurrent benign prostatic hyperplasia occurs in almost all men developing the malignancy (although the two processes are unrelated) and many others who have no evidence of cancer. Imaging evaluation of the prostate can be for diagnostic or staging purposes. While MRI can be used on occasion for diagnostic evaluation of prostatic disease, it is most often reserved for evaluation of men with known pathology. The use of MRI is important to plan the appropriate therapeutic approach to men with prostate cancer. An understanding of the newer concepts of anatomy, origins of disease, spread of cancer, and the clinical implications of pathological change of the prostate are important and are discussed in this article. The uses of MRI, the deficiencies of the study, the techniques needed to utilize the modality appropriately, and diagnostic criteria and limitations are also discussed.


Liver Kupffer cells control the magnitude of the inflammatory response in the injured brain and spinal cord.
The CNS inflammatory response is regulated by hepatic chemokine synthesis, which promotes leukocytosis and facilitates leukocyte recruitment to the site of injury. To understand the role of the individual cell populations in the liver during the hepatic response to acute brain injury, we selectively depleted Kupffer cells (KC), using clodronate-filled liposomes, and assessed the inflammatory response following a microinjection of IL-1beta into the rat brain or after a compression injury in the spinal cord. We show by immunohistochemistry that KC depletion reduces neutrophil infiltration into the IL-1beta-injected brain by 70% and by 50% into the contusion-injured spinal cord. qRT-PCR analysis of hepatic chemokine mRNA expression showed that chemokine expression in the liver after brain injury is not restricted to a single cell population. In non-depleted rats, CXCL-10, IL-1beta, CCL-2, and MIP-1alpha mRNAs were increased up to sixfold more than in KC depleted rats. However, CXCL-1 and MIP-1beta were not significantly affected by KC depletion. The reduction in chemokine mRNA expression by the liver was not associated with decreased neutrophil mobilisation as might have been expected. These findings suggest that in response to CNS injury, KC mediated mechanisms are responsible for increasing neutrophil entry to the site of CNS injury, but that neutrophil mobilisation is dependent on other non-KC mediated events. However, the suppression of KC activity may prevent secondary damage after acute brain injury.


Modifying nutritional substrates induces macrovesicular lipid droplet accumulation and metabolic alterations in a cellular model of hepatic steatosis.
Nonalcoholic fatty liver disease (NAFLD) begins with steatosis, where a mixed macrovesicular pattern of large and small lipid droplets (LDs) develops. Since in vitro models recapitulating this are limited, the aims of this study were to develop mixed macrovesicular steatosis in immortalized hepatocytes and investigate effects on intracellular metabolism by altering nutritional substrates. Huh7 cells were cultured in 11 mM glucose and 2% human serum (HS) for 7 days before additional sugars and fatty acids (FAs), either with 200 µM FAs (low fat low sugar; LFLS), 5.5 mM fructose + 200 µM FAs (low fat high sugar; LFHS), or 5.5 mM fructose + 800 µM FAs (high fat high sugar; HFHS), were added for 7 days. FA metabolism, lipid droplet characteristics, and transcriptomic signatures were investigated. Between the LFLS and LFHS conditions, there were few notable differences. In the HFHS condition, intracellular triacylglycerol (TAG) was increased and the LD pattern and distribution was similar to that found in primary steatotic hepatocytes. HFHS-treated cells had lower levels of de novo-derived FAs and secreted larger, TAG-rich lipoprotein particles. RNA sequencing and gene set enrichment analysis showed changes in several pathways including those involved in metabolism and cell cycle. Repeated doses of HFHS treatment resulted in a cellular model of NAFLD with a mixed macrovesicular LD pattern and metabolic dysfunction. Since these nutrients have been implicated in the development of NAFLD in humans, the model provides a good physiological basis for studying NAFLD development or regression in vitro.


Glycinergic Input to the Mouse Basal Forebrain Cholinergic Neurons.
The basal forebrain (BF) receives afferents from brainstem ascending pathways, which has been implicated first by Moruzzi and Magoun (1949) to induce forebrain activation and cortical arousal/waking behavior; however, it is very little known about how brainstem inhibitory inputs affect cholinergic functions. In the current study, glycine, a major inhibitory neurotransmitter of brainstem neurons, and gliotransmitter of local glial cells, was tested for potential interaction with BF cholinergic (BFC) neurons in male mice. In the BF, glycine receptor α subunit-immunoreactive (IR) sites were localized in choline acetyltransferase (ChAT)-IR neurons. The effect of glycine on BFC neurons was demonstrated by bicuculline-resistant, strychnine-sensitive spontaneous IPSCs (sIPSCs; 0.81 ± 0.25 × 10-1 Hz) recorded in whole-cell conditions. Potential neuronal as well as glial sources of glycine were indicated in the extracellular space of cholinergic neurons by glycine transporter type 1 (GLYT1)- and GLYT2-IR processes found in apposition to ChAT-IR cells. Ultrastructural analyses identified synapses of GLYT2-positive axon terminals on ChAT-IR neurons, as well as GLYT1-positive astroglial processes, which were localized in the vicinity of synapses of ChAT-IR neurons. The brainstem raphe magnus was determined to be a major source of glycinergic axons traced retrogradely from the BF. Our results indicate a direct effect of glycine on BFC neurons. Furthermore, the presence of high levels of plasma membrane glycine transporters in the vicinity of cholinergic neurons suggests a tight control of extracellular glycine in the BF.SIGNIFICANCE STATEMENT Basal forebrain cholinergic (BFC) neurons receive various activating inputs from specific brainstem areas and channel this information to the cortex via multiple projections. So far, very little is known about inhibitory brainstem afferents to the BF. The current study established glycine as a major regulator of BFC neurons by (1) identifying glycinergic neurons in the brainstem projecting to the BF, (2) showing glycine receptor α subunit-immunoreactive (IR) sites in choline acetyltransferase (ChAT)-IR neurons, (3) demonstrating glycine transporter type 2 (GLYT2)-positive axon terminals synapsing on ChAT-IR neurons, and (4) localizing GLYT1-positive astroglial processes in the vicinity of synapses of ChAT-IR neurons. The effect of glycine on BFC neurons was demonstrated by bicuculline-resistant, strychnine-sensitive spontaneous IPSCs recorded in whole-cell conditions.


Ibuprofen lacks direct antimicrobial properties for the treatment of urinary tract infection isolates.
The high incidence of urinary tract infection (UTI) among women and children, in combination with a lack of antibiotic efficacy with regard to pathogen eradication and recurrence prevention, as well as the negative side effects associated with antibiotics, has led researchers to explore the role of non-steroidal anti-inflammatory drugs as a primary management strategy. The aim of this study was to determine whether ibuprofen (IBU) or one of its major metabolites, 2-carboxyibuprofen (CIBU), could affect the growth and adhesion of the two most common uropathogens, Escherichia coli and Enterococcus faecalis. The bacterial growth and adhesion to the urothelial cells of E. coli UTI89 and E. faecalis 1131 in the presence of physiologically relevant concentrations of IBU and CIBU were assessed. The effect of IBU on bacterial adhesion to urothelial cells was also assessed following exposure to trimethoprim/sulfamethoxazole (TMP/SMX) and nitrofurantoin. Bacterial growth was not affected by IBU. Further, only at high levels of IBU not regularly found in the bladder was there a significant increase in E. faecalis 1131 attachment at growth inhibitory concentrations of TMP/SMX. There was no effect on the attachment of E. faecalis or E. coli to urothelial cells in the presence of nitrofurantoin. These studies indicate that the beneficial effects of IBU for UTI management are likely mediated through its anti-inflammatory properties rather than direct interactions with uropathogens in the bladder.


Medical students' and doctors' attitudes towards older patients and their care in hospital settings: a conceptualisation.
despite assertions in reports from governmental and charitable bodies that negative staff attitudes towards older patients may contribute to inequitable healthcare provision for older patients when compared with younger patients (those aged under 65 years), the research literature does not describe these attitudes in any detail. this study explored and conceptualised attitudes towards older patients using in-depth interviews. twenty-five semi-structured interviews with medical students and hospital-based doctors in a UK acute teaching hospital were conducted. Participants were asked about their beliefs, emotions and behavioural tendencies towards older patients, in line with the psychological literature on the definition of attitudes (affective, cognitive and behavioural information). Data were analysed thematically. attitudes towards older patients and their care could be conceptualised under the headings: (i) beliefs about older patients; (ii) older patients' unique needs and the skills required to care for them and (iii) emotions and satisfaction with caring for older patients. our findings outlined common beliefs and stereotypes specific to older patients, as opposed to older people in general. Older patients had unique needs concerning their healthcare. Participants typically described negative emotions about caring for older patients, but the sources of dissatisfaction largely related to the organisational setting and system in which the care is delivered to these patients. This study marks one of the first in-depth attempts to explore attitudes towards older patients in UK hospital settings.


Intraoperative maintenance of normoglycemia with insulin and glucose preserves verbal learning after cardiac surgery.
The hyperglycemic response to surgery may be a risk factor for cognitive dysfunction. We hypothesize that strict maintenance of normoglycemia during cardiac surgery preserves postoperative cognitive function. As part of a larger randomized, single-blind, interventional efficacy study on the effects of hyperinsulinemic glucose control in cardiac surgery (NCT00524472), consenting patients were randomly assigned to receive combined administration of insulin and glucose, titrated to preserve normoglycemia (3.5-6.1 mmol L(-1); experimental group), or standard metabolic care (blood glucose 3.5-10 mmol L(-1); control group), during open heart surgery. The patients' cognitive function was assessed during three home visits, approximately two weeks before the operation, and two months and seven months after surgery. The following tests were performed: Rey Auditory Verbal Learning Task (RAVLT for verbal learning and memory), Digit Span Task (working memory), Trail Making A & B (visuomotor tracking and attention), and the Word Pair Task (implicit memory). Questionnaires measuring specific traits known to affect cognitive performance, such as self-esteem, depression, chronic stress and social support, were also administered. The primary outcome was to assess the effect of hyperinsulinemic-normoglycemic clamp therapy versus standard therapy on specific cognitive parameters in patients receiving normoglycemic clamp, or standard metabolic care. Twenty-six patients completed the study with 14 patients in the normoglycemia and 12 patients in the control group. Multiple analysis of covariance (MANCOVA) for the RAVLT showed a significant effect for the interaction of group by visit (F = 4.07, p = 0.035), and group by visit by recall (F = 2.21, p = 0.04). The differences occurred at the second and third visit. MANCOVA for the digit span task, trail making and word pair association test showed no significant effect. Preserving intraoperative normoglycemia by intravenous insulin and glucose may prevent the impairment of memory function, both short and long-term, after cardiac surgery.


Serum lipoproteins improve after successful pharmacologic antidepressant treatment: a randomized open-label prospective trial.
Despite reports of lower plasma cholesterol in depressed patients, major depressive disorder has been shown to increase cardiovascular risk. Our objective was to study the composition of lipoproteins in depressed patients and controls and to examine the effects of pharmacologic treatment and treatment response on lipoprotein composition. Lipoprotein composition was analyzed in 65 adult inpatients at a university psychiatric hospital in Germany with DSM-IV major depressive disorder and 33 healthy controls (recruited via newspaper and radio ads) matched for age and sex. After the cross-sectional study phase, the patients were randomized in an open-label prospective trial to treatment with either mirtazapine or venlafaxine. Lipoproteins were reanalyzed after 4 weeks of treatment. Main outcome measures were total cholesterol, the low-density lipoprotein (LDL) to high-density lipoprotein (HDL) cholesterol ratio, and the LDL triglycerides to apolipoprotein B ratio. Secondary outcome measures were total triglycerides, HDL and LDL cholesterol levels, and apolipoproteins A1 and B levels. Comparisons were made between the 2 drug groups and between remitters and nonremitters as measured by the 21-item Hamilton Depression Rating Scale. The study was conducted from April 2003 through December 2007. Total cholesterol at baseline was lower in patients than in controls (mean ± SD = 4.99 ± 0.98 mmol/L vs 5.63 ± 1.01 mmol/L; P = .003), with significantly lower HDL cholesterol (P < .001) and LDL cholesterol (P = .03) in patients. However, the ratio of LDL triglycerides to apolipoprotein B, an index of size and atherogenic potential of LDL particles, was higher in depressed subjects (mean ± SD = 0.46 ± 0.14 mmol/g vs 0.38 ± 0.09 mmol/g; P = .002). Irrespective of treatment allocation, we found significant improvement of cardiovascular risk parameters in remitters but found deterioration in nonresponders. The LDL cholesterol mean change from baseline (remitters vs partial responders vs nonresponders) was -0.06 mmol/L versus +0.39 mmol/L versus +0.56 mmol/L (P = .014); the mean change in LDL/HDL cholesterol ratio was -0.50 versus +0.14 versus +0.80 (P = .002); and the mean change in the LDL triglycerides per apolipoprotein B ratio was -0.01 versus -0.01 versus +0.08 (P = .045). No drug-specific changes in lipid concentrations during treatment were observed except for total cholesterol (venlafaxine group mean = -0.02 mmol/L and mirtazapine group mean = +0.37 mmol/L; P = .033). In depressed patients, lipoprotein structure is changed toward LDL particles with a higher atherogenic potential. Remission from depression is associated with an improvement of the LDL/HDL cholesterol ratio, shifting lipoproteins toward a less atherogenic composition. Our findings should be confirmed in a larger study, as they have relevance for both researchers and clinicians. German Clinical Trial Registry Identifier: DRKS00000008.


Nonstoichiometric oxides as a continuous homologous series: linear free-energy relationship in oxygen exchange.
A novel methodology for the analysis of oxygen exchange in practically important nonstoichiometric oxides with mixed ionic electronic conductivity (MIEC) is suggested. It is based on the fact that the kinetic and thermodynamic properties of such oxides vary continuously with oxygen stoichiometry. This allows MIEC oxides to be considered as a homologous series, with the difference that traditional series are discrete in their chemical composition whereas MIEC oxides are continuous in oxygen stoichiometry. Analysis of the relations between Gibbs energies of reactions and activation barriers traditionally performed for homologous series can be useful in studies of oxygen exchange in MIEC oxides. To demonstrate the approach, thermodynamic and oxygen-exchange kinetics parameters are measured as functions of oxygen nonstoichiometry δ for two perovskites, SrCo0.8Fe0.2O3-δ and SrFeO3-δ, having metal-like and p-type semiconducting conductivities, respectively. Both oxides are shown to obey linear free energy relationships of the Brønsted-Evans-Polanyi form in spite of their different types of electronic structures. The results open up new possibilities for understanding the mechanism of the rate determining step of oxygen exchange in MIEC oxides.


Olfactory sensitivity to steroid glucuronates in Mozambique tilapia suggests two distinct and specific receptors for pheromone detection.
Cichlids offer an exciting opportunity to understand vertebrate speciation; chemical communication could be one of the drivers of African cichlid radiation. Chemical signals mediate key aspects in the lives of vertebrates and often are species specific. Dominant male Mozambique tilapia [Oreochromis mossambicus (Peters 1852)] release a sex pheromone, 5β-pregnan-3α,17α,20β-triol 3-glucuronate and its 20α-epimer, via their urine. The objective of this study was to assess the sensitivity, specificity and versatility of the olfactory system of O. mossambicus to other steroids and their conjugates using the electro-olfactogram. Oreochromis mossambicus was sensitive to several 3-glucuronidated steroids, but did not respond to prostaglandins, unconjugated steroids or 17- or 20-conjugated steroids. Stimulation of the olfactory epithelium with increasing concentrations (1 pmol l(-1) to 10 μmol l(-1)) of 5β-pregnan-3α,17α,20β-triol 3-glucuronate, 5β-pregnan-3α,17α,20α-triol 3-glucuronate, 3α,17α-dihydroxy-5β-pregnan-20-one 3-glucuronate, etiocholanolone 3α-glucuronate and 17β-estradiol 3-glucuronate produced characteristic sigmoidal concentration-response curves. However, tilapia were most sensitive to 17β-estradiol-3-glucuronate, which also had the lowest apparent EC50 and maximal response amplitude. Cross-adaptation and binary mixture experiments suggested that 5β,3α-reduced pregnan- and androstan-3-glucuronates share (a) common olfactory receptor(s), whereas 17β-estradiol 3-glucuronate is detected via (a) distinct olfactory receptor(s). In conclusion, the Mozambique tilapia has evolved high olfactory sensitivity and specificity to 3-glucuronidated steroids through two distinct olfactory receptor types; one detecting a male sex pheromone and a second detecting 17β-estradiol 3-glucuronate, a putative female-derived signal. However, O. mossambicus differs markedly in its olfactory perception from the more recently derived East African cichlid Astatotilapia burtoni, suggesting that chemical communication could, indeed, be involved in speciation.


Patient classification system evaluation: Part 2, System selection and implementation.
How can nurse executives assure optimum patient classification system performance? This two-part series advances a framework for patient classification system (PCS) evaluation. Using an expanded definition of a PCS, Part One (JONA, June) presented a discussion of the six system elements considered essential to a fully operational PCS. Part Two offers a description of PCS selection criteria as well as keys to successful system implementation. Application of this evaluation framework allows for rapid problem identification and remediation, assists with PCS selection and enhances overall system performance.


A combined pulmonary-radiology workshop for visual evaluation of COPD: study design, chest CT findings and concordance with quantitative evaluation.
The purposes of this study were: to describe chest CT findings in normal non-smoking controls and cigarette smokers with and without COPD; to compare the prevalence of CT abnormalities with severity of COPD; and to evaluate concordance between visual and quantitative chest CT (QCT) scoring. Volumetric inspiratory and expiratory CT scans of 294 subjects, including normal non-smokers, smokers without COPD, and smokers with GOLD Stage I-IV COPD, were scored at a multi-reader workshop using a standardized worksheet. There were 58 observers (33 pulmonologists, 25 radiologists); each scan was scored by 9-11 observers. Interobserver agreement was calculated using kappa statistic. Median score of visual observations was compared with QCT measurements. Interobserver agreement was moderate for the presence or absence of emphysema and for the presence of panlobular emphysema; fair for the presence of centrilobular, paraseptal, and bullous emphysema subtypes and for the presence of bronchial wall thickening; and poor for gas trapping, centrilobular nodularity, mosaic attenuation, and bronchial dilation. Agreement was similar for radiologists and pulmonologists. The prevalence on CT readings of most abnormalities (e.g. emphysema, bronchial wall thickening, mosaic attenuation, expiratory gas trapping) increased significantly with greater COPD severity, while the prevalence of centrilobular nodularity decreased. Concordances between visual scoring and quantitative scoring of emphysema, gas trapping and airway wall thickening were 75%, 87% and 65%, respectively. Despite substantial inter-observer variation, visual assessment of chest CT scans in cigarette smokers provides information regarding lung disease severity; visual scoring may be complementary to quantitative evaluation.


Diethylcarbamazine (DEC) does not induce nitric oxide (NO) synthesis.
Diethylcarbamazine (DEC) was discovered in 1947 as a potent therapeutic agent in lymphatic filariasis and has been a mainstay of antifilarial therapy over the past five decades (R. I. Hewitt, et al., 1947, Journal of Laboratory and Clinical Medicine 32, 1304-1313). Several hundred million doses of this drug have been administered to people. Despite its widespread and successful use over this prolonged time scale, its mechanism of action remains obscure (R. M. Maizels and D. A. Denham, 1992, Parasitology 105 Suppl. 549-560). Numerous studies suggest that DEC has no direct effect on the parasite (F. Hawking and W. Laurie, 1949, Lancet 2, 146-147) and that it exerts its action by stimulating host immune defense mechanisms (F. Hawking et al., 1948, Lancet 2, 730-731), or by activating host platelets to become microfilaricidal (J. Y. Cesbron et al., 1987, Nature 325(6104) 533-536). Recent data from two different laboratories suggest that NO may be involved in host defense against filarial parasites (T. V. Rajan et al., 1996, Infection and Immunity 64(8), 3351-3353; M. J. Taylor et al., 1996, Parasitology 112, 315-322). We investigated whether DEC stimulates the production of NO from murine macrophages or rat endothelial cells. DEC did not stimulate the synthesis or secretion of NO from either, nor did it synergize with interferon-gamma or tumor necrosis factor-alpha in the induction of inducible NO synthase (iNOS). In addition, there was no consistent increase in the output of inorganic nitrate, the end product of NO metabolism, in the urines of rats treated with DEC. These data suggest that DEC does not achieve its therapeutic efficacy through the induction of host iNOS.


Primary tumor resection benefited the survival of patients with distant metastatic gastric cancer.
The role of surgery in the treatment of patients with distant metastatic (M1) gastric cancer (GC) remains controversial currently. This study aimed to clarify the impact of primary tumor resection (PTR) on the survival of such patients. The surveillance, epidemiology, and end results database was adopted to extract eligible patients. We designed a retrospective case-control study. The patients were divided into two groups according to whether they received PTR. The 1:1 propensity score matching (PSM) analysis was performed to balance the confounding factors between no-surgery and surgery groups. The categorical variables were described with Chi-square tests. Cancer-specific survival (CSS) and overall survival (OS) were evaluated by Kaplan-Meier method with log-rank test. Cox proportional hazard models were utilized to identify prognostic factors of CSS. A total of 1716 patients were included, including 1108 (64.6%) patients without surgery and 608 (35.4%) patients with surgery. After PSM, most confounders were well balanced between the two comparison groups. Survival analysis in matched cohorts indicated that surgery exerted significant survival advantages in both CSS and OS curves. The median CSS was 11.0 (9.8-12.2) months in the surgery group versus 9.0 (8.0-10.0) months in the no-surgery group (P < 0.001). Multivariable Cox regression analysis identified surgery as an independent prognostic factor for favorable prognosis (hazard ratio: 0.689, 95% confidence interval: 0.538-0.881, P = 0.003). Surgery showed significant survival benefits for the patients with M1 stage GC. Our study has provided additional evidence to support PTR for these patients.


Pupillary responses to affective words in bilinguals' first versus second language.
Late bilinguals often report less emotional involvement in their second language, a phenomenon called reduced emotional resonance in L2. The present study measured pupil dilation in response to high- versus low-arousing words (e.g., riot vs. swamp) in German-English and Finnish-English late bilinguals, both in their first and in their second language. A third sample of English monolingual speakers (tested only in English) served as a control group. To improve on previous research, we controlled for lexical confounds such as length, frequency, emotional valence, and abstractness-both within and across languages. Results showed no appreciable differences in post-trial word recognition judgements (98% recognition on average), but reliably stronger pupillary effects of the arousal manipulation when stimuli were presented in participants' first rather than second language. This supports the notion of reduced emotional resonance in L2. Our findings are unlikely to be due to differences in stimulus-specific control variables or to potential word-recognition difficulties in participants' second language. Linguistic relatedness between first and second language (German-English vs. Finnish-English) was also not found to have a modulating influence.


A single nucleotide polymorphism within the novel sex-linked testis-specific retrotransposed PGAM4 gene influences human male fertility.
The development of novel fertilization treatments, including in vitro fertilization and intracytoplasmic injection, has made pregnancy possible regardless of the level of activity of the spermatozoa; however, the etiology of male-factor infertility is poorly understood. Multiple studies, primarily through the use of transgenic animals, have contributed to a list of candidate genes that may affect male infertility in humans. We examined single nucleotide polymorphisms (SNPs) as a cause of male infertility in an analysis of spermatogenesis-specific genes. We carried out the prevalence of SNPs in the coding region of phosphoglycerate mutase 4 (PGAM4) on the X chromosome by the direct sequencing of PCR-amplified DNA from male patients. Using RT-PCR and western blot analyses, we identified that PGAM4 is a functional retrogene that is expressed predominantly in the testes and is associated with male infertility. PGAM4 is expressed in post-meiotic stages, including spermatids and spermatozoa in the testes, and the principal piece of the flagellum and acrosome in ejaculated spermatozoa. A case-control study revealed that 4.5% of infertile patients carry the G75C polymorphism, which causes an amino acid substitution in the encoded protein. Furthermore, an assay for enzymatic activity demonstrated that this polymorphism decreases the enzyme's activity both in vitro and in vivo. These results suggest that PGAM4, an X-linked retrogene, is a fundamental gene in human male reproduction and may escape meiotic sex chromosome inactivation. These findings provide fresh insight into elucidating the mechanisms of male infertility.