The effect of fluoride on enamel and dentin formation in the uremic rat incisor.
Renal impairment in children is associated with tooth defects that include enamel pitting and hypoplasia. However, the specific effects of uremia on tooth formation are not known. In this study, we used rat mandibular incisors, which continuously erupt and contain all stages of tooth formation, to characterize the effects of uremia on tooth formation. We also tested the hypothesis that uremia aggravates the fluoride (F)-induced changes in developing teeth. Rats were subjected to a two-stage 5/6 nephrectomy or sham operation and then exposed to 0 (control) or 50 ppm NaF in drinking water for 14 days. The effects of these treatments on food intake, body growth rate, and biochemical serum parameters for renal function and calcium metabolism were monitored. Nephrectomy reduced food intake and weight gain. Intake of F by nephrectomized rats increased plasma F levels twofold and further decreased food intake and body weight gain. Uremia affected formation of dentin and enamel and was more extensive than the effect of F alone. Uremia also significantly increased predentin width and induced deposition of large amounts of osteodentin-like matrix-containing cells in the pulp chamber. In enamel formation, the cells most sensitive to uremia were the transitional-stage ameloblasts. These data demonstrate that intake of F by rats with reduced renal function impairs F clearance from the plasma and aggravates the already negative effects of uremia on incisor tooth development.


[Application of the characteristic interval method to differential diagnosis between the depressive phase of manic-depressive psychosis and reactive depression].
Using V. S. Genes' method of characteristic intervals 112 gradations of 36 different factors were analyzed in women under 40 years of the age with diagnoses of the depressive phase of manic-depressive psychosis (50 patients) and reactive depression (38 patients). The authors discovered 11 differential-diagnostic features relatively specific for the depressive phase of manic-depressive psychosis, and 13 features specific for reactive depression. As a result of a comprehensive analysis of these features a system for diagnostic differentiation between these conditions has been developed. This system makes it possible to diagnose correctly the depressive phase of manic-depressive psychosis in 78% of cases. In 20% of the cases the diagnostic solution remains uncertain, and in 2% erroneous.


[Results of selection for affinity in cell populations of rat transplantable tumors].
Results of selection for the affinity for lung tissue in cells of three transplanted tumors of the rat are given. Tumor cell suspensions were injected intravenously-intraperitoneally or subcutaneously-intravenously (the clonal line -- clone circuit), or intravenously-intravenously (the clone-clone circuit). The efficiency of the method has been shown in the cases of the Worker carcinoma and of a 20-methylcholantren-induced rhabdosarcoma. A 10-fold recloning of cells of the ovary ascite tumor failed to increase their affinity for the lung tissue. It is suggested that a hereditary heterogeneity with the trait "the affinity for lung tissue" may be characteristic of tumor cell populations.


Effects of interstimulus interval and contingency on classical conditioning of the Aplysia siphon withdrawal reflex.
The siphon withdrawal reflex of Aplysia undergoes differential classical conditioning with cutaneous stimulation of the siphon or mantle shelf as the discriminative conditioned stimuli (CS+ and CS-) and shock to the tail as the unconditioned stimulus (US). The reflex has proved to be useful for analyzing the neural mechanisms of conditioning. To test the generality of this experimental system, we have begun to compare the properties of conditioning in Aplysia with those of conditioning in vertebrates. We first examined the effect of the interstimulus interval (ISI) by varying the time between presentation of the CS+ and the US in different groups of animals. Significant differential conditioning was obtained when the onset of the CS+ preceded the onset of the US by 0.5 sec, and marginal conditioning was obtained when the ISI was 1.0 sec. By contrast, no significant conditioning occurred when the CS+ preceded the US by 2, 5, or 10 sec, when the onsets of the stimuli were simultaneous, or when US onset preceded the CS+ by 0.5, 1.0, or 1.5 sec (backward conditioning). We next examined the effect of contingency by giving one group of animals normal differential conditioning, and a second group the same training but with additional USs inserted between the paired trials. Presentation of these additional USs reduced the degree to which the US was contingent on the CS+, but did not change the number of pairings. Animals receiving normal training again showed significant conditioning, whereas animals receiving additional USs showed no conditioning.(ABSTRACT TRUNCATED AT 250 WORDS)


HIV-1 Escape from a Peptidic Anchor Inhibitor through Stabilization of the Envelope Glycoprotein Spike.
The trimeric HIV-1 envelope glycoprotein spike (Env) mediates viral entry into cells by using a spring-loaded mechanism that allows for the controlled insertion of the Env fusion peptide into the target membrane, followed by membrane fusion. Env is the focus of vaccine research aimed at inducing protective immunity by antibodies as well as efforts to develop drugs that inhibit the viral entry process. The molecular factors contributing to Env stability and decay need to be understood better in order to optimally design vaccines and therapeutics. We generated viruses with resistance to VIR165, a peptidic inhibitor that binds the fusion peptide of the gp41 subunit and prevents its insertion into the target membrane. Interestingly, a number of escape viruses acquired substitutions in the C1 domain of the gp120 subunit (A60E, E64K, and H66R) that rendered these viruses dependent on the inhibitor. These viruses could infect target cells only when VIR165 was present after CD4 binding. Furthermore, the VIR165-dependent viruses were resistant to soluble CD4-induced Env destabilization and decay. These data suggest that VIR165-dependent Env proteins are kinetically trapped in the unliganded state and require the drug to negotiate CD4-induced conformational changes. These studies provide mechanistic insight into the action of the gp41 fusion peptide and its inhibitors and provide new ways to stabilize Env trimer vaccines. Because of the rapid development of HIV-1 drug resistance, new drug targets need to be explored continuously. The fusion peptide of the envelope glycoprotein can be targeted by anchor inhibitors. Here we describe virus escape from the anchor inhibitor VIR165. Interestingly, some escape viruses became dependent on the inhibitor for cell entry. We show that the identified escape mutations stabilize the ground state of the envelope glycoprotein and should thus be useful in the design of stabilized envelope-based HIV vaccines.


Changing Attitudes to Alcohol in Australia.
Per-capita alcohol consumption in Australia increased between 2001 and 2007 and has subsequently declined, particularly for young people over this period. We examine trends in general attitudes to alcohol and to perceptions of risk from alcohol consumption to assess whether consumption trends have been mirrored by changing ideas about alcohol. We also examine whether trends in attitudes vary across sociodemographic subgroups. Data are from the National Drug Strategy Household Survey (n = 127,916), 2001-2013. To examine trends in attitudes to alcohol and perceptions of risk related to alcohol, logistic regression was used to assess changes over time, with interaction analyses used to explore whether changing attitudes have been consistent across sociodemographic subgroups. There were statistically significant increases in the percentage of the population naming excessive alcohol consumption as the most serious drug problem in Australia (22.6% in 2001 to 43.6% in 2013, p < .001), naming alcohol as the drug that causes the most deaths (22.7% in 2001 to 34.2% in 2013, p < .001), and suggesting thresholds for low-risk drinking below two drinks per day for men (22.4% in 2001 to 33.0% in 2013, p < .001) and women (54.4% in 2001 to 64.8% in 2013, p < .001). These trends were consistent across all sociodemographic subgroups. Australian attitudes to alcohol became more conservative between 2001 and 2013. These trends may partly explain the decline in youth drinking observed over the same period, with young people's consumption potentially more likely to be affected by growing social concern about alcohol, although shifts in attitudes occurred even while consumption was increasing.


Does Immigrant Generation Matter? Re-examining the Ethnic Density Effects on Mental Health of Ethnic Minorities in Britain.
Objectives: In this study, we extend previous research by re-examining the ethnic density effects on mental health of British ethnic minorities and exploring how the relationship varies across generations at a lower geographic scale Lower Super Output Areas (official census geographical classification designed for the consistent reporting of local statistics). Methods: We used random intercept logistic multilevel models to analyze the second wave (2010-2011) of the United Kingdom Household Longitudinal Study (UKHLS). Results: The results show that after controlling for neighborhood deprivation, respondents' socio-demographic characteristics, duration of stay in a neighborhood and moving preference, ethnic concentration has a detrimental effect on mental health for Pakistanis, Bangladeshis, black Caribbeans and black Africans. Moreover, the results show that the detrimental effects are particularly pronounced for the first-generation Pakistanis, Bangladeshis and Indians compared to their second-generation counterparts. Conclusions: Policy interventions to ameliorate ethnic disadvantages in mental health may need to be more targeted to first-generation South Asian minorities living in ethnically clustered areas, and that previous research overlooking migration generation may conceal important internal differences within British ethnic minorities.


Effects of serving high-sugar cereals on children's breakfast-eating behavior.
To test (1) whether children will consume low-sugar ready-to-eat (RTE) cereals and (2) the effects of serving high- versus low-sugar cereals on the consumption of cereal, refined sugar, fresh fruit, and milk. Using an experimental design, we randomly assigned children (n = 91) who were attending summer day camp to receive a breakfast that included either the choice of 1 of 3 high-sugar cereals (high-sugar condition) or low-sugar cereals (low-sugar condition), as well as low-fat milk, orange juice, bananas, strawberries, and sugar packets. Participants served themselves and completed a background questionnaire after eating. Researchers measured the amount and calories consumed of each food. In both conditions, children reported "liking" or "loving" the cereal they chose. Children in the low-sugar cereal condition consumed, on average, slightly more than 1 serving of cereal (35 g), whereas children in the high-sugar condition consumed significantly more (61 g) and almost twice the amount of refined sugar in total (24.4 vs 12.5 g). Milk and total calories consumed did not differ significantly between conditions, but children in the low-sugar condition were more likely to put fruit on their cereal (54% vs 8%) and consumed a greater portion of total calories from fresh fruit (20% vs 13%). Compared with serving low-sugar cereals, high-sugar cereals increase children's total sugar consumption and reduce the overall nutritional quality of their breakfast. Children will consume low-sugar cereals when offered, and they provide a superior breakfast option.


Genetic diversity of hepatitis B virus strains derived worldwide: genotypes, subgenotypes, and HBsAg subtypes.
Sequences of 234 complete genomes and 631 hepatitis B surface antigen genes were used to assess the worldwide diversity of hepatitis B virus (HBV). Apart from the described two subgenotypes each for A and F, also B, C, and D divided into four subgenotypes each in the analysis of complete genomes supported by significant bootstrap values. The subgenotypes of B and C differed in their geographical distribution, with B1 dominating in Japan, B2 in China and Vietnam, B3 confined to Indonesia, and B4 confined to Vietnam, all strains specifying subtype ayw1. Subgenotype C1 was common in Japan, Korea, and China; C2 in China, South-East Asia, and Bangladesh, and C3 in the Oceania comprising strains specifying adrq-, and C4 specifying ayw3 is encountered in Aborigines from Australia. This pattern of defined geographical distribution was less evident for D1-D4, where the subgenotypes were widely spread in Europe, Africa, and Asia, possibly due to their divergence having occurred a longer time ago than for genotypes B and C, with D4 being the first split and still the dominating subgenotype of D in the Oceania. The genetic diversity of HBV and the geographical distribution of its subgenotypes provide a tool to reconstruct the evolutionary history of HBV and may help to complement genetic data in the understanding of the evolution and past migrations of man.


Survival of cats with naturally occurring chronic renal failure: effect of dietary management.
Fifty cats with naturally occurring stable chronic renal failure (CRF) were entered into a prospective study on the effect of feeding a veterinary diet restricted in phosphorus and protein with or without an intestinal phosphate binding agent on their survival from initial diagnosis. Twenty-nine cats accepted the veterinary diet, whereas compliance (due to limited intake by the cats or owner resistance to diet change) was not achieved in the remaining 21. At diagnosis, both groups of cats were matched in terms of age, bodyweight, plasma creatinine, phosphate, potassium and parathyroid hormone (PTH) concentrations, packed cell volume and urine specific gravity. Feeding the veterinary diet was associated with a reduction in plasma phosphate and urea concentrations and prevented the increase in plasma PTH concentrations seen in cats not receiving the diet. Cats fed the veterinary diet survived for longer when compared with those that were not (median survival times of 633 versus 264 days). These data suggest that feeding a diet specifically formulated to meet the needs of cats with CRF, together with phosphate binding drugs if required, controls hyperphosphataemia and secondary renal hyperparathyroidism, and is associated with an increased survival time.


The role of pituitary gonadotropins in gonadal sex differentiation in the protogynous Malabar grouper, Epinephelus malabaricus.
The aim of this study was to clarify the roles of 2 gonadotropins (GTHs), follicle-stimulating hormone (FSH) and luteinizing hormone (LH), on sex differentiation in the protogynous Malabar grouper, Epinephelus malabaricus. To do this, the mRNA expression patterns of GTH subunits (cga, fshb, and lhb) in the fish pituitary throughout gonadal sex differentiation were investigated. Real-time reverse transcriptase (RT)-PCR showed that cga and fshb were present in the undifferentiated and ovarian differentiation stages, and that the expression levels significantly increased after ovarian differentiation (AOD). However, lhb was not expressed before ovarian differentiation (BOD) and was first detected AOD. Next, to investigate the differentiation and distribution of Fshb and Lhb-producing cells in the pituitary of fish throughout gonadal sex differentiation, immunohistochemical analysis was used to detect teleost GTH subunits. Positive immunoreactivity against Fshb and Lhb was not detected in the pituitary BOD; Fshb and Lhb-positive cells first appeared in the pituitary AOD. It therefore seems unlikely that pituitary gonadotropins play a major role in the control of gonadal sex differentiation in the Malabar grouper.


Predictors for early introduction of solid food among Danish mothers and infants: an observational study.
Early introduction of complementary feeding may interfere with breastfeeding and the infant's self-controlled appetite resulting in increased growth. The aim of the present study was to investigate predictors for early introduction of solid food. In an observational study Danish mothers filled in a self-administered questionnaire approximately six months after birth. The questionnaire included questions about factors related to the infant, the mother, attachment and feeding known to influence time for introduction of solid food. The study population consisted of 4503 infants. Data were analysed using ordered logistic regression models. Outcome variable was time for introduction to solid food. Almost all of the included infants 4386 (97%) initiated breastfeeding. At weeks 16, 17-25, 25+, 330 infants (7%); 2923 (65%); and 1250 (28%), respectively had been introduced to solid food. Full breastfeeding at five weeks was the most influential predictor for later introduction of solid food (OR = 2.52 CI: 1.93-3.28). Among infant factors male gender, increased gestational age at birth, and higher birth weight were found to be statistically significant predictors. Among maternal factors, lower maternal age, higher BMI, and being primipara were significant predictors, and among attachment factors mother's reported perception of the infant as being temperamental, and not recognising early infant cues of hunger were significant predictors for earlier introduction of solid food. Supplementary analyses of interactions between the predictors showed that the association of maternal perceived infant temperament on early introduction was restricted to primiparae, that the mother's pre-pregnancy BMI had no impact if the infant was fully breastfed at week five, and that birth weight was only associated if the mother had reported early uncertainty in recognising infant's cues of hunger. Breastfeeding was the single most powerful indicator for preventing early introduction to solid food. Modifiable predictors pointed to the importance of supporting breastfeeding and educating primipara and mothers with low birth weight infants to be able to read and respond to their infants' cues to prevent early introduction to solid food.


Laboratory detection methods for the human coronaviruses.
Coronaviruses are a group of envelop viruses which lead to diseases in birds and mammals as well as human. Seven coronaviruses have been discovered in humans that can cause mild to lethal respiratory tract infections. HCoV-229E, HCoV-OC43, HCoV-NL63, and HCoV-HKU1 are the low-risk members of this family and the reason for some common colds. Besides, SARS-CoV, MERS-CoV, and newly identified SARS-CoV-2, which is also known as 2019-nCoV, are the more dangerous viruses. Due to the rapid spread of this novel coronavirus and its related disease, COVID-19, a reliable, simple, fast, and low-cost detection method is necessary for patient diagnosis and tracking worldwide. Human coronaviruses detection methods were classified and presented in this article. The laboratory detection techniques include RT-PCR, RT-LAMP, electrochemical and optical biosensors for RNA detection, and whole virus or viral proteins detection assays.


Multiple myeloma and B cell lymphoma. Investigation of IL-6, IL-6 receptor antagonist (IL-6RA), and GP130 antagonist (GP130A) using various parameters in an in vitro model.
Interleukin-6 (IL-6) affects the survival and proliferation of myeloma cells via autocrine and/or paracrine mechanisms. In this study, we investigated the effects of IL-6, IL-6 receptor antagonist (IL-6RA), and gp130 antagonist (gp130A) on the membrane expressions of IL-6R and gp130, on the viability, on the proliferation, on the DNA synthesis, and on the cell cycle phases in several multiple myeloma (MM) cell lines and B cell lymphoma cell lines. Our results showed that (1) all five MM cell lines (OPM-2, RPMI-8226, U-266, KMS-12-BM, MOLP-8) expressed surface IL-6R and gp130, the B cell lymphomas (WSU-1, DOHH-2, U-698) expressed only gp130; (2) exogenous IL-6 markedly up-regulated the expression of membrane IL-6R (up to 186%) and down-regulated the gp130 receptor (down to 4%) in MM cell lines, the membrane expression of gp130 in B cell lymphomas was not altered; (3) IL-6 markedly increased the spontaneous proliferation (up to 151%) in all MM cell lines, that of B cell lymphomas was not affected; (4) IL-6 increased the DNA synthesis in the S cell cycle phase of MM cells and arrested the stage G2/M, IL-6 was ineffective in any cell cycle phase of B cell lymphoma; (5) IL-6RA inhibited the membrane IL-6R partially, the proliferation was decreased only slightly; and (6) although gp130A inhibited the membrane gp130 completely, the proliferation was decreased 81-78% in MM and B cell lymphoma cell lines. This means that gp130 is not absolutely necessary for the cellular signalling cascade via JAK/STAT and RAS/MAPK pathways involved in proliferation and viability. Our results give an indication in the therapy of MM: IL-6 antibody (IL-6A) alone or in combination with IL-6RA. The latter could be more effective. This kind of therapy is not recommended for B cell lymphoma, as these cells have no IL-6R.


Morphologic and mitogenic responses of rabbit synovial fibroblasts to transforming growth factor beta require transforming growth factor alpha or epidermal growth factor.
We tested the hypothesis that normal synovial fibroblasts might proliferate in response to transforming growth factors (TGFs), peptides that are extracted with acid-ethanol from bovine kidney or salivary gland and that cause anchorage-independent growth of normal cells. A 72-hour exposure of confluent monolayers of rabbit synovial fibroblasts in 10% fetal calf serum to partially purified TGF-beta in the presence of TGF-alpha gave a 2- to 5-fold increase in incorporation of 3H-thymidine, protein content, and cell number. Similar results were obtained with high pressure liquid chromatography-purified TGF-beta in the presence of epidermal growth factor (EGF) (a type of TGF-alpha). By itself, purified TGF-beta was not mitogenic; it depended absolutely on EGF. However, only TGF-beta along with EGF, and not EGF alone, induced a marked morphologic change: a piling up of cells into foci resembling those commonly seen in primary cultures of rheumatoid synovial cells. Mitogenic responses induced by the TGF-beta-EGF combination were prevented by all-trans-retinoic acid but not by indomethacin or dexamethasone. The data indicate that TGF-beta, a peptide extracted from normal cells, can act in concert with EGF to cause proliferation and piling up of synovial cells and raise the possibility that these factors may play a role in rheumatoid arthritis and other proliferative but nonmalignant diseases as well.


Thrombolytic therapy in the setting of arterial line-induced ischemia.
To evaluate the use of percutaneous transcatheter thrombolysis in the treatment of thrombosis due to radial artery cannulation. Seven patients (4 women; age range 41-62 years) with symptomatic cannulation-induced thrombosis and failure to improve after systemic anticoagulation underwent 8 catheter-directed thrombolytic infusions at our institution over a 3-year period. Access was either antegrade through the brachial artery or retrograde through the femoral artery. Thrombolytic infusions with urokinase began 2 to 12 days (average 6) after removal of the radial artery catheter. The thrombolytic infusion was successful in 5 of 7 patients based on angiographic flow restoration with <20% residual thrombus and significant clinical improvement in the ischemia. When systemic anticoagulation has failed, percutaneous catheter-directed thrombolytic infusion appears to be effective in the treatment of most patients with severe ischemic hand symptoms due to thrombosis after radial artery cannulation.


Psychometric Properties of a Spanish-Language Version of a Short-Form FAMCARE: Applications to Caregivers of Patients With Alzheimer's Disease and Related Dementias.
Although family satisfaction is recognized as a critical indicator of quality care for persons with serious illness, Spanish-language measures are limited. The study aims were to develop a Spanish translation of the short-form Family Satisfaction With End-of-Life Care (FAMCARE), investigate its psychometric properties in Hispanic caregivers to patients with Alzheimer's disease and related dementias (ADRD; N = 317; 209 interviewed in Spanish), and add parameters to an existing item bank. Based on factor analyses, the measure was found to be essentially unidimensional. Reliabilities from a graded item response theory model were high; the average estimate was 0.93 for the total and Spanish-language subsample. Discrimination parameters were high, and the model fit adequate. This is the first study to examine the performance of the short-form FAMCARE measure among Hispanics and caregivers to patients with ADRD. The short-form measure can be recommended for Hispanics and caregivers to patients with ADRD.


Low Body Mass Index Is Associated with Higher Odds of COPD and Lower Lung Function in Low- and Middle-Income Countries.
The relationship of body mass index (BMI) with lung function and COPD has been previously described in several high-income settings. However, few studies have examined this relationship in resource-limited settings where being underweight is more common. We evaluated the association between BMI and lung function outcomes across 14 diverse low- and middle-income countries. We included data from 12,396 participants aged 35-95 years and used multivariable regressions to assess the relationship between BMI with either COPD and lung function while adjusting for known risk factors. An inflection point was observed at a BMI of 19.8 kg/m2. Participants with BMI < 19.8 kg/m2 had a 2.28 greater odds (95% CI 1.83-2.86) of having COPD and had a 0.21 (0.13-0.30) lower FEV1 and 0.34 (0.27-0.41) lower FEV1/FVC z-score compared to those with BMI ≥ 19.8 kg/m2. The association with lung function remained even after excluding participants with COPD. Individuals with lower BMI were more likely to have COPD and had lower lung function compared to those in higher BMI. The association with lung function remained positive even after excluding participants with COPD, suggesting that being underweight may also play a role in having worse lung function.


A STUDY OF RABBIT GAMMA-GLOBULIN ALLOTYPY BY MEANS OF HETEROIMMUNIZATIONS.
Representatives of two species, the chicken and the goat, were immunized by injection with Freund's adjuvant, of specific precipitates prepared from the antisera of rabbits with various allotypic specificities. The precipitation reactions of the resulting antisera with normal rabbit sera were studied in liquid and in gellified media. In these reactions the isotypic specificity and often one or the other of the allotypic specificities of rabbit gamma-globulin were involved. It was always easier to obtain antibodies against the specificities controlled by the alleles of locus b (Ab4, Ab5, Ab6) than of locus a (Aa1, Aa2, Aa3). The precipitation reaction of antibodies against a specificity of the a series could always be inhibited by an excess (nearly always moderate) of a serum lacking this specificity. A similar inhibition of the antibodies against the specificities of the b series was often impossible, especially if the antiserum had been collected after a second injection of immunizing material. The reactions of 58 Ab4(+) sera with a goat anti-Ab4 serum and of 28 Ab5(+) sera with a chicken anti-AbS serum were studied in gel tubes (simple diffusion). The constant observation of two precipitation zones due respectively to two types of molecules, Ab4(+) and Ab4(-) (or Ab5(+) and Ab5(-)), indicated the existence of an appreciable number of Ab4(-) or Ab5(-) molecules, even in supposed homozygotes. In the latter rabbits Ab4(-) or Ab5(-) molecules are b(-) molecules; i.e., molecules lacking the specificities of the b series. Measurements of the distances between the two zones in the reaction of 72 sera revealed significant differences between sera of individuals of the same phenotype, as well as systematic differences not only between sera of homozygotes and of heterozygotes, but also between heterozygotes of different genotypes. These measurements were used to evaluate the ratio of concentrations of the two types of molecules detected by each of the two antisera. The occurrence of b(-) molecules in every one of the fairly large number of sera studied indicates a new aspect of the heterogeneity of gamma-globulin. Although detected by means of allotypy, this heterogeneity is not dependent upon factors which vary with individuals. These results are discussed in connection with what is known of the structure of gamma-globulin.


[Effect of extra-intracranial microvascular anastomoses on the evolution of stenoses in the internal carotid artery system].
Clinical and angiographic examination of 20 patients with stenoses in the system of the internal carotid artery before and after operations for the formation of extra-intracranial microvascular anastomoses (EICMA), and during EICMA functioning demonstrated the evolution of the stenoses. It was characterized both by their increase (even up to the occurrence of thrombosis) and by their regression. No evolution of the stenoses occurred when EICMA failed to function. The character of evolution of the stenosis was determined by its localization. The trend of the evolution was dependent on the etiology of the stenosis. The degree of EICMA functioning differed and was determined by the character of stenosis evolution. With the development of stenosis into thrombosis, the neurological symptom complex was characterized both by marked augmentation followed by regression as well as by the absence of any changes or positive dynamics of the neurological changes.


Cannabinoid receptor 2-mediated attenuation of CXCR4-tropic HIV infection in primary CD4+ T cells.
Agents that activate cannabinoid receptor pathways have been tested as treatments for cachexia, nausea or neuropathic pain in HIV-1/AIDS patients. The cannabinoid receptors (CB(1)R and CB(2)R) and the HIV-1 co-receptors, CCR5 and CXCR4, all signal via Gαi-coupled pathways. We hypothesized that drugs targeting cannabinoid receptors modulate chemokine co-receptor function and regulate HIV-1 infectivity. We found that agonism of CB(2)R, but not CB(1)R, reduced infection in primary CD4+ T cells following cell-free and cell-to-cell transmission of CXCR4-tropic virus. As this change in viral permissiveness was most pronounced in unstimulated T cells, we investigated the effect of CB(2)R agonism on to CXCR4-induced signaling following binding of chemokine or virus to the co-receptor. We found that CB(2)R agonism decreased CXCR4-activation mediated G-protein activity and MAPK phosphorylation. Furthermore, CB(2)R agonism altered the cytoskeletal architecture of resting CD4+ T cells by decreasing F-actin levels. Our findings suggest that CB(2)R activation in CD4+ T cells can inhibit actin reorganization and impair productive infection following cell-free or cell-associated viral acquisition of CXCR4-tropic HIV-1 in resting cells. Therefore, the clinical use of CB(2)R agonists in the treatment of AIDS symptoms may also exert beneficial adjunctive antiviral effects against CXCR4-tropic viruses in late stages of HIV-1 infection.


Activation of Resolution Pathways to Prevent and Fight Chronic Inflammation: Lessons From Asthma and Inflammatory Bowel Disease.
Formerly considered as a passive process, the resolution of acute inflammation is now recognized as an active host response, with a cascade of coordinated cellular and molecular events that promotes termination of the inflammatory response and initiates tissue repair and healing. In a state of immune fitness, the resolution of inflammation is contained in time and space enabling the restoration of tissue homeostasis. There is increasing evidence that poor and/or inappropriate resolution of inflammation participates in the pathogenesis of chronic inflammatory diseases, extending in time the actions of pro-inflammatory mechanisms, and responsible in the long run for excessive tissue damage and pathology. In this review, we will focus on how resolution can be the target for therapy in "Th1/Th17 cell-driven" immune diseases and "Th2 cell-driven" immune diseases, with inflammatory bowel diseases (IBD) and asthma, as relevant examples. We describe the main cells and mediators stimulating the resolution of inflammation and discuss how pharmacological and dietary interventions but also life style factors, physical and psychological conditions, might influence the resolution phase. A better understanding of the impact of endogenous and exogenous factors on the resolution of inflammation might open a whole area in the development of personalized therapies in non-resolving chronic inflammatory diseases.


Spotlight on agalsidase beta in Fabry disease.
Agalsidase beta (Fabrazyme) is a recombinant human alpha-galactosidase A enzyme approved for intravenous use in the treatment of Fabry disease. Fabry disease is a progressive, multisystemic, potentially life-threatening disorder caused by a deficiency of alpha-galactosidase A. This deficiency results in accumulation of glycosphingolipids, particularly globotriaosylceramide (GL-3), in the lysosomes of various tissues. This accumulation is the underlying driver of disease progression. Agalsidase beta provides an exogenous source of alpha-galactosidase A. Intravenous agalsidase beta is effective and well tolerated in patients with Fabry disease. In a phase III trial, agalsidase beta was shown to clear GL-3 from various target cells and, in a subsequent extension of this trial, prevent GL-3 reaccumulation. In a post-approval trial, agalsidase beta was shown to provide significant clinical benefit by reducing the risk of a major clinical event. Thus, agalsidase beta represents an important advance in the treatment of Fabry disease, and agalsidase beta therapy should be strongly considered in patients with Fabry disease who are suitable candidates.


The Occurrence of Malignancy in Trypanosoma brucei brucei by Rapid Passage in Mice.
Pleomorphic Trypanosoma brucei are best known for their tightly controlled cell growth and developmental program, which ensures their transmissibility and host fitness between the mammalian host and insect vector. However, after long-term adaptation in the laboratory or by natural evolution, monomorphic parasites can be derived. The origin of these monomorphic forms is currently unclear. Here, we produced a series of monomorphic trypanosome stocks by artificially syringe-passage in mice, creating snapshots of the transition from pleomorphism to monomorphism. We then compared these artificial monomorphic trypanosomes, alongside several naturally monomorphic T. evansi and T. equiperdum strains, with the pleomorphic T. brucei. In addition to failing to generate stumpy forms in animal bloodstream, we found that monomorphic trypanosomes from laboratory and nature exhibited distinct differentiation patterns, which are reflected by their distinct differentiation potential and transcriptional changes. Lab-adapted monomorphic trypanosomes could still be induced to differentiate, and showed only minor transcriptional differences to that of the pleomorphic slender forms but some accumulated differences were observed as the passages progress. All naturally monomorphic strains completely fail to differentiate, corresponding to their impaired differentiation regulation. We propose that the natural phenomenon of trypanosomal monomorphism is actually a malignant manifestation of protozoal cells. From a disease epidemiological and evolutionary perspective, our results provide evidence for a new way of thinking about the origin of these naturally monomorphic strains, the malignant evolution of trypanosomes may raise some concerns. Additionally, these monomorphic trypanosomes may reflect the quantitative and qualitative changes in the malignant evolution of T. brucei, suggesting that single-celled protozoa may also provide the most primitive model of cellular malignancy, which could be a primitive and inherent biological phenomenon of eukaryotic organisms from protozoans to mammals.


Learning Spectral Fractional Anisotropy and Mean Diffusivity Features as Neuroimaging Biomarkers for Tracking White Matter Integrity Changes in Myotonic Dystrophy Type 1 Patients using Deep Convolutional Neural Networks.
Myotonic dystrophy type 1 (DM1) is a genetic neuromuscular progressive multisystem disease that results in a broad spectrum of clinical central nervous system (CNS) involvement, including problems with memory, attention, executive functioning, and social cognition. Fractional anisotropy and mean diffusivity along-tract data calculated using diffusion tensor imaging techniques play a vital role in assessing white matter microstructural changes associated with neurodegeneration caused by DM1. In this work, a novel spectrogram-based deep learning method is proposed to characterize white matter network alterations in DM1 with the goal of building a deep learning model as neuroimaging biomarkers of DM1. The proposed method is evaluated on fractional anisotropies and mean diffusivities along-tract data calculated for 25 major white matter tracts of 46 DM1 patients and 96 unaffected controls. The evaluation data consists of a total of 7100 spectrogram images. The model achieved 91% accuracy in identifying DM1, a significant improvement compared to previous methods.Clinical relevance- Clinical care of DM1 is particularly challenging due to DM1 multisystem involvement and the disease variability. Patients with DM1 often experience neurological and psychological symptoms, such as excessive sleepiness and apathy, that greatly impact their quality of life. Some of DM1 CNS symptoms may be responsive to treatment. The goal of this research is to gain a deeper understanding of the impact of DM1 on the CNS and to develop a deep learning model that can serve as a biomarker for the disease, with the potential to be used in future clinical trials as an outcome measure.


Cognition, but not sensation, mediates age-related changes in the ability to monitor the environment.
The objective of the study was to determine which age-related changes in sensation and cognition are associated with age-related changes in the ability to monitor the environment. To that end, a proxy measure of the ability to monitor the environment (useful field of view, UFOV) and measures of sensation and cognition were collected from young adult (N = 61) and older adult subjects (N = 79). Although UFOV performance was expected to be mediated primarily by cognition rather than by sensation, it was somewhat unexpected to find no reliable associations between UFOV and sensory functioning beyond those of age and cognition.


Numerical method to optimize the polar-azimuthal orientation of infrared superconducting-nanowire single-photon detectors.
A finite-element method for calculating the illumination-dependence of absorption in three-dimensional nanostructures is presented based on the radio frequency module of the Comsol Multiphysics software package (Comsol AB). This method is capable of numerically determining the optical response and near-field distribution of subwavelength periodic structures as a function of illumination orientations specified by polar angle, φ, and azimuthal angle, γ. The method was applied to determine the illumination-angle-dependent absorptance in cavity-based superconducting-nanowire single-photon detector (SNSPD) designs. Niobium-nitride stripes based on dimensions of conventional SNSPDs and integrated with ~ quarter-wavelength hydrogen-silsesquioxane-filled nano-optical cavity and covered by a thin gold film acting as a reflector were illuminated from below by p-polarized light in this study. The numerical results were compared to results from complementary transfer-matrix-method calculations on composite layers made of analogous film-stacks. This comparison helped to uncover the optical phenomena contributing to the appearance of extrema in the optical response. This paper presents an approach to optimizing the absorptance of different sensing and detecting devices via simultaneous numerical optimization of the polar and azimuthal illumination angles.


Self-Perception of Changes in Routines in Adults and Older Adults Associated to Social Distancing Due to COVID-19-A Study in São Paulo, Brazil.
COVID-19 is an acute respiratory illness with higher mortality in older adults. This condition is spread person-to-person through close contact, and among policies employed to decrease transmission are the improvement of hygiene habits and physical distancing. Although social distancing has been recognized as the best way to prevent the transmission, there are concerns that it may promote increased depression symptoms risk and anxiety, mainly in older adults. This cross-sectional study aimed to verify self-concept of social distancing in adults compared to older adults. All participants, over 18 years and residents of São Paulo state (Brazil), were invited to join this research study by a message application and answered an interdisciplinary questionnaire during the period from May 23 to June 23, 2020. The questions were divided into the following aspects: sociodemographic data, financial conditions, routine-related perception, perception of health, physical and emotional state, and eating habits. The younger adult group was composed of 139 participants, with a mean age of 43.15 years (±10.92), and the older adult group was composed of 437 participants with a mean age of 67.59 years (±6.13) of both sex. Changes in routine during the period of social distance were reported by 95% of adults and 96.8% of older adults, but adults indicated more significant alterations in routine. Although there was no difference between groups for several aspects, adults revealed greater alterations in sleep quality, evacuation frequency, and more difficulty to perform daily activities at home. Further studies are necessary to follow up the impacts of social distancing among adults and older adults in different socioeconomic contexts to better understand the long-term alterations and the necessity of interventions.


Anisotropy effect of three-membered rings in (1)H NMR spectra: quantification by TSNMRS and assignment of the stereochemistry.
The spatial magnetic properties (through space NMR shieldings, TSNMRSs) of cyclopropane; of the heteroanalogous oxirane, thiirane, and aziridine; and of various substituted mono-, dis-, and tris-cyclic analogues have been computed by the GIAO perturbation method employing the nucleus independent chemical shift (NICS) concept and visualized as iso-chemical-shielding surfaces (ICSSs) of various size and direction. The TSNMRS values, thus obtained, can be employed to visualize the anisotropy (ring current) effect of the cyclopropane ring moiety. This approach has been employed to qualify and quantify substituent influences and contributions of appropriate ring heteroatoms O, NH, and S on the anisotropy (ring current) effect of three-membered ring moieties, and to assign the stereochemistry of mono-, bis-, and tris-cyclic structures containing cyclopropane as a structural element. Characteristic examples are included.


Community-based football in men with prostate cancer: 1-year follow-up on a pragmatic, multicentre randomised controlled trial.
Physical exercise has been shown to be effective in relation to fatigue, aerobic fitness, and lower body strength in men with prostate cancer. However, research into the clinically relevant effects of interventions conducted in heterogeneous patient populations and in real-life clinical practice settings is warranted. We conducted a pragmatic, multicentre, parallel randomised controlled trial in 5 Danish urological departments. Recruitment began in May 2015, the first participant was randomised in June 2015, and the last participant was included in February 2017. In total, 214 men with prostate cancer were randomly assigned to either 6 months of free-of-charge football training twice weekly at a local club (football group [FG]) (n = 109) or usual care (usual care group [UG]) (n = 105), including brief information on physical activity recommendations at randomisation. Participants were on average 68.4 (SD 6.2) years old, 157 (73%) were retired, 87 (41%) were on castration-based treatment, 19 (9%) had received chemotherapy, and 41 (19%) had skeletal metastases at baseline. In this 1-year follow-up study, we evaluated the effects of community-based football training on the following outcomes: primary outcome, quality of life; secondary outcomes: continuation of football after 6 months, hip and lumbar spine bone mineral density (BMD), mental health score, fat and lean body mass, and safety outcomes, i.e., fractures, falls, and hospital admissions. Intention to treat (ITT) and per protocol (PP) analyses were conducted. No statistically significant between-group difference was observed in change in prostate-cancer-specific quality of life (ITT: 1.9 points [95% CI -1.9 to 5.8], p = 0.325; PP: 3.6 points [95% CI -0.9 to 8.2], p = 0.119). A statistically significant between-group difference was observed in change in total hip BMD, in favour of FG (0.007 g/cm2 [95% CI 0.004 to 0.013], p = 0.037). No differences were observed in change in lumbar spine BMD or lean body mass. Among patients allocated to football, 59% chose to continue playing football after the end of the 6-month intervention period. At 1-year follow-up in the PP population, FG participants had more improvement on the Mental Component Summary (2.9 [95% CI 0.0 to 5.7], p = 0.048 points higher) than UG participants, as well as a greater loss of fat mass (-0.9 kg [95% CI -1.7 to -0.1], p = 0.029). There were no differences between groups in relation to fractures or falls. Hospital admissions were more frequent in UG compared to FG (33 versus 20; the odds ratio based on PP analysis was 0.34 for FG compared to UG). There were 3 deaths in FG and 4 in UG. Main limitations of the study were the physically active control group and assessment of physical activity by means of self-report. In this trial, participants allocated to football appeared to have improved hip BMD and fewer hospital admissions. Men who played football more than once a week for 1 year lost fat mass and reported improved mental health. Community-based football proved to be acceptable, even when club membership was not subsidised. ClinicalTrials.gov NCT02430792.


Quercetin-induced inhibition and synergistic activity with cisplatin - a chemotherapeutic strategy for nasopharyngeal carcinoma cells.
Nasopharyngeal carcinoma (NPC) is a unique tumour of epithelial origin with a distinct geographical distribution, genetic predisposition and environmental as well as dietary influence as aetiological factors. Standard NPC treatment regimes, such as radiotherapy and concurrent chemotherapy with cytotoxic drugs, can produce undesirable complications often associated with significant toxicity. Here, we report the effects of a widely distributed flavonoid, quercetin, on cell proliferation, apoptosis and cell cycle arrest. The effects of combining quercetin and cisplatin on human NPC cells were explored. Cell proliferation was monitored by the dynamic, impedance-based cell analyzer (xCELLigence system) and the MTS assay. Ki67 proliferation antigen and fatty acid synthase (FASN) level was examined by Western blotting. Flow cytometry was also carried out to study the effects of quercetin on cell cycle and apoptosis status. At 100 μM, quercetin inhibited cell proliferation and decreased expression of FASN and Ki67 antigen. Cell cycle analysis revealed a substantial increase in the proportion of cells in the G2/M phase. We also demonstrated the enhanced cytotoxic effects of quercetin treatment in concomitant with the chemotherapeutic drug, cisplatin, in cultured NPC cells. The combination index (CI) value of quercetin-cisplatin combination was < 1, indicating synergism. Our study showed that quercetin exhibited synergistic effects with cisplatin against NPC cells. Dose-reduction index (DRI) values > 1 implied the possibility of reducing the cisplatin dosage required to treat NPC, with the addition of quercetin. In turn, this could reduce the risk of cisplatin-associated toxicity. The potential of combining quercetin with cisplatin as a chemotherapeutic strategy for treatment of NPC should be explored further.


Prevalence and risk factors for aspirin and clopidogrel resistance in cerebrovascular stenting.
The prevalence of antiplatelet drug resistance among patients who undergo cerebrovascular stent placement is unknown. We aimed to assess the feasibility of monitoring antiplatelet drug effects in a single-center cohort undergoing cerebrovascular stent placement. We prospectively collected medical, laboratory, and radiographic data on patients who underwent cerebrovascular stent placement. We used the rapid platelet function assay-aspirin (RPFA-ASA) to calculate aspirin reaction units (ARU) and the P2Y12 assay to calculate P2Y12 reaction units and percentage platelet inhibition. Aspirin resistance was defined as ARU > 550, whereas clopidogrel resistance was defined as percentage platelet inhibition < 40%. Among 76 patients, stent indications were the following: wide-neck aneurysm (57, 75.0%), symptomatic intracranial stenosis (12, 15.7%), carotid stenosis (5, 6.6%), and vertebral stenosis (2, 2.6%). For aspirin, the median dosage per week was 1300 mg and median ARU was 410. Among 71 patients on aspirin, 3 patients (4.2%) were resistant; there was a significant inverse correlation between aspirin dose and ARU (r = -0.31, P = .01). Among 55 patients on clopidogrel, the median dosage per week was 525 mg with a mean platelet inhibition of 43.2%. Twenty-eight patients (51.9%) were clopidogrel-resistant. In a multivariable linear regression model, age older than 55 years (b = -16.3, P = .020) and diabetes (b = -26.8, P = .015) were inversely related to percentage platelet inhibition. Using point-of-care tests, we found that aspirin resistance is relatively uncommon, whereas clopidogrel resistance occurred in half of patients undergoing cerebrovascular stent placement. Further studies should focus on ideal doses, timing, and duration of antiplatelet therapy for cerebrovascular stent placement.


Relation between Breast Cancer and High Glycemic Index or Glycemic Load: A Meta-analysis of Prospective Cohort Studies.
Breast cancer is the commonest form of cancer in women worldwide. It has been suggested that chronic hyperinsulinemia associated with insulin resistance plays a role in breast cancer etiology. To test the hyperinsulinemia hypothesis, a dietary pattern associated with a high glycemic index and glycemic load, both proxies for chronic hyperinsulinemia, should be associated with an increased risk of breast cancer. A meta-analysis restricted to prospective cohort studies was undertaken using a random effects model with tests for statistical significance, publication bias and heterogeneity. The metric for analysis was the risk of breast cancer in the highest relative to the lowest glycemic index and glycemic load dietary pattern. A dietary pattern with a high glycemic index was associated with a summary relative risk (SRR) of 1.05 (95% CI: 1.00, 1.11), and a high glycemic load with a SRR of 1.06 (95% CI: 1.00, 1.13). Adjustments for body mass index [BMI], physical activity and other lifestyle factors did not influence the SRR, nor did menopausal status and estrogen receptor status of the tumor. In conclusion, the current evidence supports a modest association between a dietary pattern with high glycemic index or glycemic load and the risk of breast cancer.


Gangliosides block Aggregatibacter Actinomycetemcomitans leukotoxin (LtxA)-mediated hemolysis.
Aggregatibacter actinomycetemcomitans is an oral pathogen and etiologic agent of localized aggressive periodontitis. The bacterium is also a cardiovascular pathogen causing infective endocarditis. A. actinomycetemcomitans produces leukotoxin (LtxA), an important virulence factor that targets white blood cells (WBCs) and plays a role in immune evasion during disease. The functional receptor for LtxA on WBCs is leukocyte function antigen-1 (LFA-1), a β-2 integrin that is modified with N-linked carbohydrates. Interaction between toxin and receptor leads to cell death. We recently discovered that LtxA can also lyse red blood cells (RBCs) and hemolysis may be important for pathogenesis of A. actinomycetemcomitans. In this study, we further investigated how LtxA might recognize and lyse RBCs. We found that, in contrast to a related toxin, E. coli α-hemolysin, LtxA does not recognize glycophorin on RBCs. However, gangliosides were able to completely block LtxA-mediated hemolysis. Furthermore, LtxA did not show a preference for any individual ganglioside. LtxA also bound to ganglioside-rich C6 rat glioma cells, but did not kill them. Interaction between LtxA and C6 cells could be blocked by gangliosides with no apparent specificity. Gangliosides were only partially effective at preventing LtxA-mediated cytotoxicity of WBCs, and the effect was only observed when a high ratio of ganglioside:LtxA was used over a short incubation period. Based on the results presented here, we suggest that because of the similarity between N-linked sugars on LFA-1 and the structures of gangliosides, LtxA may have acquired the ability to lyse RBCs.


Influence of nerve on the formation and survival of acetylcholine receptor and cholinesterase patches on embryonic Xenopus muscle cells in culture.
Embryonic Xenopus muscle cells grown in culture develop discrete patches of high acetylcholine receptor (AChR) density. By following identified muscle cells after staining with fluorescent alpha-bungarotoxin, we have found that many of these AChR patches survive in a fixed position for several days. For AChR patches on the lower surface of the cell (the surface apposed to the culture dish), more than 60% of those which were followed beginning on day 2 survived for a further 4 days. The survival rate was greater when patches were followed from day 3 or later and was almost as high for AChR patches on the upper surface. New AChR patches also formed on all of the muscle cells. When muscle cells were cultured together with spinal cord cells, nerve-muscle contacts developed with a characteristic localization of AChRs along the path of contact. AChR patches did not form elsewhere on these contacted cells. Nerve-contacted muscle cells examined 2 to 3 days after adding spinal cord cells to established (2- to 5-day-old) muscle cultures also exhibited a marked reduction of AChR patches away from the site of contact. This reduction was not due to the nerve having contacted pre-existing AChR patches. Rather, the findings indicate that contact by an appropriate nerve inhibits the formation of AChR patches elsewhere on the contracted muscle cells and reduces the survival of pre-existing AChR patches. Nerve contact also inhibited the formation of cholinesterase (ChE) patches remote from the site of contact and appeared to cause some reduction in the survival of pre-existing ChE patches. Spontaneous twitching was not observed in these experiments, thereby indicating that the remote effects of nerve contact were not mediated by muscle action potentials or contraction. Such remote influences of the nerve may play a role in determining the pattern of innervation on individual muscle cells.


The frequency of reduction loss after arthroscopic fixation of acute acromioclavicular dislocations using a double-button device, and its effect on clinical and radiological results.
The aim of this study was to investigate the effect of reduction loss of more than 3 mm on clinical and radiological results after at least 2 years of follow-up after arthroscopic fixation of acute acromioclavicular joint dislocations using a double-button device. Thirty-six patients who had acute (< 3 weeks old), type III or V acromioclavicular (AC) joint dislocations underwent arthroscopic fixation of the AC joint using a double-button device. Clinical and radiological evaluations were performed at preoperative, postoperative first day, 3 months and last follow-up. When the coracoclavicular (CC) distances of patients at the last follow-up were compared to the early postop CC distances, those with a difference of 3 mm or less were grouped as group A and those with a difference of more than 3 mm were grouped as group B. There was no statistically significant difference between the groups in terms of age, gender, follow-up time, time from injury to surgery, return to work, and distribution of Rockwood classification. Pre-operative CC distance was reduced from 18.7 ± 3.5 to 8.5 ± 0.6 in the early postoperative period. Anatomic reduction was achieved in all patients compared with the unaffected side (CC distance 8.6 ± 0.7). However, the CC distance increased to 9.9 ± 1.5 at the third-month follow-up and increased to 11 ± 2.7 at the last follow-up. There were no significant Constant score differences between the groups in the preoperative and last follow-up periods (p > 0.05). At the last follow-up, the mean Acromioclavicular Joint Instability (ACJI) score of group A was 84.4 ± 8, whereas it was 68.3 ± 8.3 for group B, and the difference was statistically significant (p < 0.01). Furthermore, the subjective evaluation and aesthetic subjective satisfaction values of group B were lower than group A (p < 0.01). Reduction loss of more than 3 mm was observed in 25% of patients after arthroscopic fixation of acute acromioclavicular dislocations using a double-button device. Although this loss did not create a statistically significant difference in Constant scores, AC joint-specific tests such as ACJI, subjective evaluation, and aesthetic subjective satisfaction values were significantly impaired.


Factors affecting reproductive performance of dairy cows.
We conducted two studies to determine how herd management practices and traits of individual cows affect performance of the herd and of the cow within a herd. Management practices, reproductive performance of the herd, and relationships between management and reproductive performance were characterized on 83 dairy farms with 7596 cows. Data included 21 management variables (e.g., facilities, herd health program, estrous detection program) and 8 performance variables obtained from Dairy Herd Improvement or unofficial records (e.g., size of herd, production, days open). Although varying among herds, annual average herd incidences of reproductive disorders and reproductive performance were similar to those reported. Managerial practices influenced incidences of retained placenta and uterine infection, days open of cows not bred and of all cows, services per conception, and percentages of herd open more than 100 days and culled for low production. Veterinarian was the most consistent variable influencing herd reproductive performance. Data also were collected from production and lifetime records of 2532 cows in 19 herds. Reproductive performance was affected by season of calving, production, maturity, and reproductive disorders. Several cows with extremely poor reproductive records were maintained.


Liver Transplantation Outcomes in Recipients with High Model for End-Stage Liver Disease (MELD) Scores: The Relevance of MELD Scores.
BACKGROUND The aim of this study was to assess risk factors for postoperative mortality after liver transplantation among patients with Model for End-Stage Liver Disease (MELD) scores ≥35, with special focus on the MELD scores. MATERIAL AND METHODS Data from 68 primary liver transplantations in patients with MELD scores ≥35 among 1376 liver transplantations performed in the Department of General, Transplant, and Liver Surgery (Medical University of Warsaw) between January 2002 and October 2014 were analyzed retrospectively. Postoperative (90-day) mortality was set as the primary outcome measure. RESULTS Postoperative mortality was 29.4% (20 of 68). The overall survival rates after 1, 5, and 10 years were 61.9%, 59.7%, and 59.7%, respectively. According to univariate analyses, MELD (p=0.014), conventional technique of liver transplantation (p=0.049), intraoperative fresh frozen plasma (p=0.040), and red blood cells (p=0.026) transfusions were risk factors for postoperative mortality. MELD score was the only independent risk factor for postoperative mortality (p=0.023) in multivariate analysis. According to receiver operating characteristics analysis, the optimal cut-off for MELD score in prediction of postoperative mortality was ≥43 (Area Under Curve=0.703, 95% Confidence Interval 0.575-0.831). Postoperative mortality was 21.4% and 42.3% among patients with MELD score <43 and ≥43, respectively (p=0.066). CONCLUSIONS MELD score is an important predictor of early mortality after liver transplantation, even among recipients with high MELD scores. In particular, patients with MELD score ≥43 should be considered as very high-risk candidates for liver transplantation.


Marcanine G, a new cytotoxic 1-azaanthraquinone from the stem bark of Goniothalamus marcanii Craib.
The ethanolic extract from the stem bark of Goniothalamus marcanii Craib was shown in preliminary brine shrimp lethality data having good cytotoxic activity. Further bioassay guided isolation was done by means of solvent partition, chromatography and precipitation to provide four isolated compounds: a novel compound 1 with the core structure of 1-azaanthraquinone moiety referred as marcanine G; as well as compounds 2-4 with known aristolactam structures namely, piperolactam C, cepharanone B and taliscanine. These compounds were characterised by spectroscopic techniques. The assessment of cytotoxicity was established on an SRB assay using doxorubicin as a positive control. Marcanine G (1) was considered the most active compound indicating the IC50 values of 14.87 and 15.18 μM against human lung cancer cells (A549) and human breast cancer cells (MCF7), respectively. However, 2 showed mild activity with the IC50 values of 83.72 and 82.32 μM against A549 and MCF7 cells, respectively.


Factors associated with suicide mortality among reproductive age women: a case-control study.
Examine demographic, psychosocial, pregnancy-related, and healthcare utilisation factors associated with suicide mortality among reproductive age women. Data from nine health care systems in the Mental Health Research Network were included. A case-control study design was used in which 290 reproductive age women who died by suicide (cases) from 2000 to 2015 were matched with 2,900 reproductive age women from the same healthcare system who did not die by suicide (controls). Conditional logistic regression was used to analyse associations between patient characteristics and suicide. Women of reproductive age who died by suicide were more likely to have mental health (aOR = 7.08, 95% CI: 5.17, 9.71) or substance use disorders (aOR = 3.16, 95% CI: 2.19, 4.56) and to have visited the emergency department in the year prior to index date (aOR = 3.47, 95% CI: 2.50, 4.80). Non-Hispanic White women (aOR = 0.70, 95% CI: 0.51, 0.97) and perinatal (pregnant or postpartum) women were less likely to have died by suicide (aOR = 0.27, 95% CI: 0.13, 0.58). Reproductive age women with mental health and/or substance use disorders, prior emergency department encounters, or who are of racial or ethnic minority status were at increased risk of suicide mortality and may benefit from routine screening and monitoring. Future research should further examine the relationship between pregnancy-related factors and suicide mortality.


Ethical Shades of Gray: International Frequency of Scientific Misconduct and Questionable Research Practices in Health Professions Education.
To maintain scientific integrity and engender public confidence, research must be conducted responsibly. Whereas deliberate scientific misconduct such as data fabrication is clearly unethical, other behaviors-often referred to as questionable research practices (QRPs)-exploit the ethical shades of gray that color acceptable practice. This study aimed to measure the frequency of self-reported misconduct and QRPs in a diverse, international sample of health professions education (HPE) researchers. In 2017, the authors conducted an anonymous, cross-sectional survey study. The web-based survey contained 43 items that asked respondents to rate how often they had engaged in a variety of irresponsible research behaviors. The items were adapted from previously published surveys. In total, 590 HPE researchers took the survey. The mean age was 46 years (SD = 11.6), and the majority of participants were from the United States (26.4%), Europe (23.2%), and Canada (15.3%). The three most frequently reported irresponsible research behaviors were adding authors who did not qualify for authorship (60.6%), citing articles that were not read (49.5%), and selectively citing papers to please editors or reviewers (49.4%). Additionally, respondents reported misrepresenting a participant's words (6.7%), plagiarizing (5.5%), inappropriately modifying results (5.3%), deleting data without disclosure (3.4%), and fabricating data (2.4%). Overall, 533 (90.3%) respondents reported at least one irresponsible behavior. Notwithstanding the methodological limitations of survey research, these findings indicate that a substantial proportion of HPE researchers report a range of misconduct and QRPs. Consequently, reforms may be needed to improve the conduct of HPE research.


Molecular subclassification determined by human papillomavirus and epidermal growth factor receptor status is associated with the prognosis of oropharyngeal squamous cell carcinoma.
Human papillomavirus (HPV) infection is an indicator of good response to chemoradiotherapy in oropharyngeal squamous cell carcinoma (OPSCC), and epidermal growth factor receptor (EGFR) is a molecular-therapeutic target in head and neck squamous cell carcinoma. Here we investigated the prevalence and prognostic significance of HPV infection and EGFR alteration in OPSCC. We analyzed the presence of high-risk HPV using in situ hybridization, protein expressions of p16 and EGFR using immunohistochemistry, and the EGFR gene copy number gain using chromogenic in situ hybridization (CISH) in 105 cases of OPSCC. The biopsy specimens before chemoradiotherapy were used for these analyses. HPV infection and p16 protein overexpression were detected in 53.3% and 52.4% of the OPSCCs, and each factor was associated with better overall survival (P = .0026 and P = .0026) and nonkeratinizing histology (P = .0002 and P = .0004), respectively. EGFR gene copy number gain (high polysomy or amplification) was detected in 12.4% of the OPSCCs and was correlated with EGFR protein overexpression (P = .0667) and worse overall survival (P < .0001). HPV infection and EGFR gene copy number gain (EGFR CISH positive) were mutually exclusive. The HPV-negative/EGFR CISH-positive OPSCCs had significantly worse overall survival than did the HPV-positive/EGFR CISH-negative OPSCCs and HPV-negative/EGFR CISH-negative OPSCCs (P < .0001 and P < .0001, respectively). The EGFR CISH-negative OPSCCs had favorable prognosis irrespective of HPV infection. Our results suggest that EGFR gene copy number gain-positive tumors represent an HPV-negative, aggressive subgroup of OPSCCs. The molecular subclassification of OPSCCs based on HPV infection and EGFR status may serve as important information for appropriate therapeutic strategy.


Methamphetamine decreases CD4 T cell frequency and alters pro-inflammatory cytokine production in a model of drug abuse.
The reason co-morbid methamphetamine use and HIV infection lead to more rapid progression to AIDS is unclear. We used a model of methamphetamine self-administration to measure the effect of methamphetamine on the systemic immune system to better understand the co-morbidity of methamphetamine and HIV. Catheters were implanted into the jugular veins of male, Sprague Dawley rats so they could self-administer methamphetamine (n=18) or be given saline (control; n=16) for 14 days. One day after the last operant session, blood and spleens were collected. We measured serum levels of pro-inflammatory cytokines, intracellular IFN-γ and TNF-α, and frequencies of CD4(+), CD8(+), CD200(+) and CD11b/c(+) lymphocytes in the spleen. Rats that self-administered methamphetamine had a lower frequency of CD4(+) T cells, but more of these cells produced IFN-γ. Methamphetamine did not alter the frequency of TNF-α-producing CD4(+) T cells. Methamphetamine using rats had a higher frequency of CD8(+) T cells, but fewer of them produced TNF-α. CD11b/c and CD200 expression were unchanged. Serum cytokine levels of IFN-γ, TNF-α and IL-6 in methamphetamine rats were unchanged. Methamphetamine lifetime dose inversely correlated with serum TNF-α levels. Our data suggest that methamphetamine abuse may exacerbate HIV disease progression by activating CD4 T cells, making them more susceptible to HIV infection, and contributing to their premature demise. Methamphetamine may also increase susceptibility to HIV infection, explaining why men who have sex with men (MSM) and frequently use methamphetamine are at the highest risk of HIV infection.


Enemy-free space and habitat-specific host specialization in a butterfly.
The majority of herbivorous insects have relatively specialized food habits. This suggests that specialization has some advantage(s) over generalization. Traditionally, feeding specialization has been thought to be linked to digestive or other food-related physiological advantages, but recent theory suggests that generalist natural enemies of herbivorous insects can also provide a major selective pressure for restricted host plant range. The European swallowtail butterfly Papilio machaon utilizes various plants in the Apiaceae family as hosts, but is an ecological specialist being monophagous on Angelica archangelica in southern Sweden. This perennial monocarp grows in three seaside habitat types: (1) on the barren rocky shore in the absence of any surrounding vegetation, (2) on the rocky shore with some surrounding vegetation, and (3) on species-rich meadows. The rocky shore habitat harbors few invertebrate generalist predators, whereas a number of invertebrate predators abound in the meadowland habitat. Here, we test the importance of enemy-free space for feeding specialization in Papilio machaon by assessing survival of larvae placed by hand on A. archangelica in each of the three habitat types, and by assessing the habitat-specificity of adult female egg-laying behavior by recording the distribution of eggs laid by free-flying adult females among the three habitat types. Larval survival was substantially higher in the rocky shore habitat than in the meadowland and significantly higher on host plants without surrounding vegetation on the rocky shore. Eggs laid by free-flying females were found in all three habitat types, but were significantly more frequent in the rocky shore habitat, suggesting that females prefer to lay eggs in the habitat type where offspring survival is highest. These results show that larval survivorship on the same host plant species can be strongly habitat-specific, and suggest that enemy-free space is an underlying factor that drives feeding specialization in Papilio machaon.


Validity of the activPAL3 activity monitor in people moderately affected by Multiple Sclerosis.
Walking is the primary form of physical activity performed by people with Multiple Sclerosis (MS), therefore it is important to ensure the validity of tools employed to measure walking activity. The aim of this study was to assess the criterion validity of the activPAL3 activity monitor during overground walking in people with MS. Validity of the activPAL3 accelerometer was compared to video observation in 20 people moderately affected by MS. Participants walked 20-30m twice along a straight quiet corridor at a comfortable speed. Inter-rater reliability of video observations was excellent (all intraclass correlations >0.99). The mean difference (activPAL3- mean of raters) was -4.70±9.09, -4.55s±10.76 and 1.11s±1.11 for steps taken, walking duration and upright duration respectively. These differences represented 8.7%, 10.0% and 1.8% of the mean for each measure respectively. The activPAL3 tended to underestimate steps taken and walking duration in those who walked at cadences of ≤38 steps/min by 60% and 47%, respectively. The activPAL3 is valid for measuring walking activity in people moderately affected by MS. It is accurate for upright duration regardless of cadence. In participants with slow walking cadences, outcomes of steps taken and walking duration should be interpreted with caution.


Lysergic acid diethylamide (LSD) promotes social behavior through mTORC1 in the excitatory neurotransmission.
Clinical studies have reported that the psychedelic lysergic acid diethylamide (LSD) enhances empathy and social behavior (SB) in humans, but its mechanism of action remains elusive. Using a multidisciplinary approach including in vivo electrophysiology, optogenetics, behavioral paradigms, and molecular biology, the effects of LSD on SB and glutamatergic neurotransmission in the medial prefrontal cortex (mPFC) were studied in male mice. Acute LSD (30 μg/kg) injection failed to increase SB. However, repeated LSD (30 μg/kg, once a day, for 7 days) administration promotes SB, without eliciting antidepressant/anxiolytic-like effects. Optogenetic inhibition of mPFC excitatory neurons dramatically inhibits social interaction and nullifies the prosocial effect of LSD. LSD potentiates the α-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA) and 5-HT2A, but not N-methyl-D-aspartate (NMDA) and 5-HT1A, synaptic responses in the mPFC and increases the phosphorylation of the serine-threonine protein kinases Akt and mTOR. In conditional knockout mice lacking Raptor (one of the structural components of the mTORC1 complex) in excitatory glutamatergic neurons (Raptorf/f:Camk2alpha-Cre), the prosocial effects of LSD and the potentiation of 5-HT2A/AMPA synaptic responses were nullified, demonstrating that LSD requires the integrity of mTORC1 in excitatory neurons to promote SB. Conversely, in knockout mice lacking Raptor in GABAergic neurons of the mPFC (Raptorf/f:Gad2-Cre), LSD promotes SB. These results indicate that LSD selectively enhances SB by potentiating mPFC excitatory transmission through 5-HT2A/AMPA receptors and mTOR signaling. The activation of 5-HT2A/AMPA/mTORC1 in the mPFC by psychedelic drugs should be explored for the treatment of mental diseases with SB impairments such as autism spectrum disorder and social anxiety disorder.


Risk Factors for Workplace Bullying: A Systematic Review.
Objective: The goal of this study was to systematically review risk factors for workplace bullying. Methods: The search was carried out in two databases. Studies with estimates of risk factors for workplace bullying were included in the review. We assessed the quality of the selected studies using an adapted version of the Downs and Black checklist. Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) and Meta-analyses of Observational Studies in Epidemiology (MOOSE) guidelines were used for reporting papers. Results: Fifty-one papers were included, and 70.6% were from European countries. Women were reported to be at higher risk of being bullied in most studies (odds ratio (OR) from 1.17 to 2.77). Authoritarian and laissez-faire leadership styles were positively associated with bullying. Several psychosocial factors, such as stress (OR from 1.37 to 4.96), and occupational risks related to work organization, such as flexible work methods, role conflict, role ambiguity, monotonous or rotating tasks, high demands, pressure of work, and unclarity of duties were strongly associated with bullying. Discussion: The findings highlight the central role of organizational factors in bullying. Policies to prevent bullying must address the culture of organizations, facing the challenge of developing a new management and leadership framework.


A low-affinity vasopressin V2-receptor gene in a kindred with X-linked nephrogenic diabetes insipidus.
In this study, a mutation in vasopressin Type 2 receptor (V2R) in a patient with hereditary nephrogenic diabetes insipidus (NDI) has been identified and characterized. The sequencing of the V2R gene from the patient revealed that there was a missense mutation (TAT to TGT) resulting in the substitution of 205Tyr for Cys in the putative third extracellular domain. The expression analysis in COS cells showed that the binding affinity of the mutant receptor (KD = 19.8 nM) for arginine vasopressin was much lower than that of the wild-type receptor (KD = 1.8 nM) so that intracellular cAMP production stimulated by arginine vasopressin was impaired in cells with the mutant V2R. From these results, it was concluded that the single amino-acid substitution of V2R is responsible for this familial disease.


The prevalence of hypertension and its associated risk factors in a newly developed country.
The aim of the present study was to determine the prevalence of hypertension and cardiovascular disease (CVD) risk factors and its association with the sociodemographic, behavioral, and lifestyle characteristics of the adult qatari population. This is a cross-sectional study, which was carried out in primary health care clinics (PHCs). The survey was conducted from January through to July 2003 among qatari nationals 25-65-years of age. Of the 1500 patients who were approached to participate in study, 1208 (80.5%) gave their consent. Face-to-face interviews were based on a questionnaire that included variables on age, sex, socioeconomic status (SES), parity, income level, cigarette smoking, physical activity, lifestyle, body mass index (BMI), and blood pressure. Hypertension was defined according to the world health organization criteria as systolic blood pressure (SBP) >140 mm Hg or diastolic blood pressure (DBP) >90 mm Hg, or both. Overall, the prevalence of hypertension (BP >/=140 or >/=90 mm Hg, or both or known hypertensive) was 32.1% (32.6% in men and 31.7% in women). The age-standardized prevalence of hypertension (adjusted to the adult population of Qatar) was 31.1% (95% confidence interval 26.7-35.5%) in men and 30.2% in women (95% confidence interval 25.8-34.6%). The CVD risk factor of obesity was more prominent among women 528 (78.3%) than among men 334 (68.9%) (p<0.001). Physical inactivity was again predominant among women and found to be highly significant. A stepwise logistic regression analysis revealed that diabetes (p<0.0001); hormonal problems (p<0.0001), consumption of animal fats (p<0.0001), risk of heart disease (p<0.0001), and BMI (p=0.0307) were all associated with hypertension. The present study was directed at determining the prevalence and risk factors of high blood pressure in the adult population of Qatar. The characterization of these factors will contribute to defining more effective and specific strategies to screen for and control hypertension and CVD.


[Methods of research on group therapy].
In the field of what is known as group talk therapy it is rather difficult, but absolutely necessary to do more than merely discuss various concepts and make occasional observations. The possible methods described include the "pre" and "post" systems of principles, practices, and procedures; the reference group method; the method of investigating process variables; and the analog experimental method. Examples are given for each of the above methods. It is evident from a number of theoretical designs that major achievements will finally come from a suitable combination of different methods of research.


CCK-8, CCK-4 and gastrin-induced contractions in guinea pig ileum: evidence for differential release of acetylcholine and substance P by CCK-A and CCK-B receptors.
The cholecystokinin (CCK) receptor involved in contraction of guinea pig ileal longitudinal muscle to cholecystokinin is poorly understood; some studies have suggested that contraction was mediated via a CCK-A receptor whereas other studies have implicated CCK-B receptors in ileal contraction to CCK. To clarify this, we compared the effects of CCK-8 sulfate, CCK-4 and gastrin in radioligand binding studies and longitudinal ileal contractility in vitro. Contraction to all three peptides was abolished by tetrodotoxin (3 x 10(-7)M), confirming the neuronal nature of the CCK receptors mediating contraction to all three peptides. Maximal CCK-8S contractions were inhibited by 80% in the presence of atropine (10(-6)M), and entirely by the combination of atropine and a substance P receptor antagonist (3 x 10(-5)M). CCK-4 and gastrin-induced contractions were unaffected by substance P receptor blockade, but were abolished by atropine. Two selective CCK-A and CCK-B receptor antagonists, L-364,718 and L-365,260, respectively, were used to probe further the receptors involved in ileal contraction to this peptide family. Radioligand binding studies in mouse brain, rat pancreas and guinea pig stomach confirmed the selectivity of these antagonists. The CCK-A selective antagonist, L-364,718, potently inhibited ileal contractions to CCK-8S (-log KB = 9.35) with 10-fold lower affinity at receptors mediating contraction to CCK-4 (-log KB = 8.25). In contrast, the CCK-B receptor antagonist, L-365,260, did not affect contraction to CCK-8S (-log KB less than 7) but potently inhibited contraction to CCK-4 (-log KB = 9.24).(ABSTRACT TRUNCATED AT 250 WORDS)


Effect of reducing the phytate content and of partially hydrolyzing the protein in soy formula on zinc and copper absorption and status in infant rhesus monkeys and rat pups.
Although soy formulas have been designed to meet the nutrient requirements of human infants, they also contain phytate, which may negatively affect trace element absorption. We evaluated the effect of removing phytate on zinc and copper absorption and status in infant rhesus monkeys and suckling rat pups and evaluated differences between intact and partially hydrolyzed soy protein. In monkeys, regular and low-phytate soy formulas were fed exclusively for 4 mo and whole-body absorption and retention of 65Zn, 67Cu, 59Fe, 54Mn, and 47Ca were determined at different time points with a whole-body counter. Subsequently, zinc and copper absorption from several human infant formulas and the effect of phytate concentration were evaluated in suckling rat pups by using 65Zn and 64Cu. Finally, infant rhesus monkeys were fed low-phytate formulas with intact or hydrolyzed soy protein for 4 mo and plasma zinc and copper were measured monthly. In the first monkey study, zinc absorption at 1 mo was higher from low-phytate soy formula (36%) than from regular soy formula (22%), whereas there was no significant difference between groups in the absorption of other minerals. Plasma copper was significantly lower in monkeys fed low-phytate soy formula from 2 to 4 mo. In rat pups, zinc absorption was significantly higher from low-phytate soy formula (78%) than from regular soy formula (51%) and hydrolysis of the protein had no significant effect. Phytate content or protein hydrolysis did not significantly affect copper absorption. In the second monkey study, plasma copper concentrations were highest in monkeys fed the low-phytate, hydrolyzed-protein soy formula. Reducing the phytate content and partially hydrolyzing the protein in soy formula had a beneficial effect on zinc and copper absorption and status in infant rhesus monkeys.


A ubiquitin-proteasome inhibitor bortezomib suppresses the expression of CYP11B2, a key enzyme of aldosterone synthesis.
CYP11B2 is a key enzyme involved in the synthesis of the mineralocorticoid aldosterone. CYP11B2 expression in the adrenal glands is controlled by the renin-angiotensin system (RAS), and plays an important role in the maintenance of electrolyte metabolism in higher organisms. Abnormal overexpression of CYP11B2 results in the disruption of mineral balance and can lead to hypertension. Though the molecular mechanism of the regulation of CYP11B2 expression has remained elusive, we hypothesize that compounds that prevent CYP11B2 expression could represent a novel class of antihypertensive drugs. In this study, we established a high-throughput screening system to identify such compounds, and subsequently carried out chemical screening. We found that the ubiquitin-proteasome inhibitor bortezomib could suppress CYP11B2 expression and secretion of aldosterone induced by angiotensin II (Ang II) in adrenocortical H295R cells. Moreover, bortezomib down-regulated the Cyp11b2 mRNA expression facilitated in the adrenal gland of Tsukuba hypertensive mice, resulting in subsequent lowering of their blood pressures. Furthermore, we observed the characteristic alteration of H3K27ac in the adrenal CYP11B2 gene promoter induced by Ang II stimulation, which was found to be disrupted by bortezomib. Taken together, these results suggest the possibility of developing novel antihypertensive drugs that prevent CYP11B2 expression.


The dynamic DNA methylation cycle from egg to sperm in the honey bee Apis mellifera.
In honey bees (Apis mellifera), the epigenetic mark of DNA methylation is central to the developmental regulation of caste differentiation, but may also be involved in additional biological functions. In this study, we examine the whole genome methylation profiles of three stages of the haploid honey bee genome: unfertilised eggs, the adult drones that develop from these eggs and the sperm produced by these drones. These methylomes reveal distinct patterns of methylation. Eggs and sperm show 381 genes with significantly different CpG methylation patterns, with the vast majority being more methylated in eggs. Adult drones show greatly reduced levels of methylation across the genome when compared with both gamete samples. This suggests a dynamic cycle of methylation loss and gain through the development of the drone and during spermatogenesis. Although fluxes in methylation during embryogenesis may account for some of the differentially methylated sites, the distinct methylation patterns at some genes suggest parent-specific epigenetic marking in the gametes. Extensive germ line methylation of some genes possibly explains the lower-than-expected frequency of CpG sites in these genes. We discuss the potential developmental and evolutionary implications of methylation in eggs and sperm in this eusocial insect species.


The development and pilot evaluation of a nutrition education intervention programme for pregnant teenage women (food for life).
A healthy diet in pregnancy is important for both maternal and infant health but this may be difficult to achieve particularly for groups such as teenage pregnant women, many of whom are from disadvantaged backgrounds. To our knowledge this is the first report of a practical nutrition education programme for this group in the UK. An intervention was designed incorporating seven informal food preparation sessions, which allowed opportunities for discussion of nutritional, and other topics (e.g. food safety and well-being in pregnancy). Midwives in a community centre setting led the sessions. The acceptability of the package to participants and midwives was recorded and pre- and post-intervention data collected on sociodemographic details, dietary intake (using an eating-habits questionnaire and a 24-h dietary recall) and cooking skills. The midwives found the package easy to follow and use. The 16 (of the 120 invited) women who attended found the courses helpful but objective evaluation of dietary intake was not possible because of poor compliance. The nutrition education programme was favourably received by midwives and the women who participated. However recruitment was problematic and alternative methods of delivering and evaluating such a package should be investigated.


[Cytotoxicity of sera against brain and spinal cord antigens in relation to lymphocytes of varying origin].
Rabbit sera against antigens prepared from the brain and the spinal cord antigens were investigated in a cytotoxic test with mouse and guinea pig lymphocytes. None of the sera exerted a cytotoxic effect on the bone marrow lymphocytes. The sera against mouse brain and spinal cord and guinea pig brain and myelin isolated from it exerted the greatest cytotoxic activity; the cytotoxicity was maximum against the thymocytes, less pronounced against the lymph node lymphocytes, and least--against the spleen cells. The cytotoxicity of the sera against the bovine spinal cord homogenate, myelin and the basic protein isolated from it was the minimal and equal with lymphocytes from any of the three mentioned sources. The serum against the encephalitogenic polypeptide 2c was practically devoid of the cytotoxic activity. The encephalitogenic activity of the 2c fraction was greater than that of the myelin and basic protein from the bovine spinal cord. Experiment of antibrain serum absorption suggested that the brain cortex contained a cross-reacting antigen. The subcutaneous injection of a relatively high dose (224 x 10(6)) of thymocytes in the complete Freund's adjuvant failed to induce the development of allergic encephalomyelitis in guinea pigs.


Genetic study on the mandibular molar size of rats.
Tooth size is determined by genetic and environmental factors like other quantitative characters such as body weight and body height. However, the degree of the relative contribution of both factors to the determination of tooth size has not been well clarified. In order to study the genetic and environmental factors affecting tooth size, we carried out a diallel cross mating by the cohabitation of pairs of males and females among 10 strains of rats. The bucco-lingual widths of the first, second, and third molars of the right mandible were measured in each offspring of F1 population. The body weight was also measured as a parameter that might indicate systemic growth factor in connection with tooth development. The quantitative genetic analysis was performed based on Wearden's model (Heredity 19:669-680, 1964). As a result, the size of the first and the second molars was more significantly controlled by genetic effect than maternal effect, while maternal effect could not be ignored for the size of the third molar in addition to genetic effect. The genetic effect on body weight became greater with age, while the maternal effect showed its maximum influence upon the body weight around the weaning. It is concluded that the size of the molar teeth beginning to develop in the uterus and to be calcified just after birth was mainly controlled by genetic factor, and that the size of the molar teeth beginning to develop approximately after birth was mainly controlled by maternal effect affecting body weight at the same period.


Overexpression of protein phosphatase 4 correlates with poor prognosis in patients with stage II pancreatic ductal adenocarcinoma.
Protein phosphatase 4 (PP4) has been reported to be overexpressed in breast and lung cancers. PP4 plays an important role in the regulation of centrosome maturation, DNA repair, NF-κB, and c-jun-NH(2)-kinase (JNK) signaling pathways. However, the expression and functions of PP4 in pancreatic cancer have not been studied. We examined the expression of PP4 catalytic subunit (PP4C) protein in 133 patients with stage II pancreatic ductal adenocarcinoma (PDAC) and their paired benign pancreatic samples (N = 113) by immunohistochemistry. To confirm the immunohistochemical results, we measured PP4C protein and mRNA levels by Western blotting and real-time reverse transcriptase PCR. Using univariate and multivariate analysis, we correlated PP4C expression with survival and other clinicopathologic features. PP4C was overexpressed in 75 of 133 (56.4%) stage II PDAC samples, which was significantly higher than the paired benign pancreatic tissue (15%, 17 of 113). PP4C mRNA expression levels were also higher in PDAC samples than the paired benign pancreatic tissue. Overexpression of PP4C in PDAC samples was associated with higher frequencies of distant metastasis (P = 0.02) and poor disease-free and overall survivals in patients with stage II PDAC (P = 0.006 and 0.02) independent of tumor size, margin status, and lymph node status (stage). Our study showed that PP4C is overexpressed in PDAC. Overexpression of PP4C in PDAC samples is associated with poor prognosis in patients with stage II PDAC. Therefore, targeting PP4 signaling pathway may represent a new approach for the treatment of PDAC. Our study showed that PP4C is an independent prognostic factor in patients with stage II PDAC.


Evidence for a mutualistic relationship between the cyanobacteria Nostoc and fungi Aspergilli in different environments.
Symbiotic partnerships are widespread in nature and in industrial applications yet there are limited examples of laboratory communities. Therefore, using common photobionts and mycobionts similar to those in natural lichens, we create an artificial lichen-like symbiosis. While Aspergillus nidulans and Aspergillus niger could not obtain nutrients from the green algae, Chlorella, and Scenedesmus, the cyanobacteria Nostoc sp. PCC 6720 was able to support fungal growth and also elevated the accumulation of total biomass. The Nostoc-Aspergillus co-cultures grew on light and CO2 in an inorganic BG11 liquid medium without any external organic carbon and fungal mycelia were observed to peripherally contact with the Nostoc cells in liquid and on solid media at lower cell densities. Overall biomass levels were reduced after implementing physical barriers to indicate that physical contact between cyanobacteria and heterotrophic microbes may promote symbiotic growth. The synthetic Nostoc-Aspergillus nidulans co-cultures also exhibited robust growth and stability when cultivated in wastewater over days to weeks in a semi-continuous manner when compared with axenic cultivation of either species. These Nostoc-Aspergillus consortia reveal species-dependent and mutually beneficial design principles that can yield stable lichen-like co-cultures and provide insights into microbial communities that can facilitate sustainability studies and broader applications in the future. KEY POINTS: • Artificial lichen-like symbiosis was built with wild-type cyanobacteria and fungi. • Physical barriers decreased biomass production from artificial lichen co-cultures. • Artificial lichen adapted to grow and survive in wastewater for 5 weeks.


The C terminus of mouse monocyte chemoattractant protein 1 (MCP1) mediates MCP1 dimerization while blocking its chemotactic potency.
The extracellular protease plasmin cleaves mouse MCP1 (monocyte chemoattractant protein 1) at lysine 104, releasing a 50-amino acid C-terminal domain. The cleavage event increases the chemotactic activity of MCP1 and, by doing so, promotes the progression of excitotoxic injury in the central nervous system in pathological settings. The mechanism through which the cleavage event enhances MCP1-mediated chemoattraction is unknown; to investigate it, we use wild-type and mutant forms of recombinant MCP1. Full-length MCP1 (FL-MCP1) is secreted by cells as a dimer or multimer. We show that a mutant truncated at the C terminus, K104Stop-MCP1, does not dimerize, revealing that the C terminus mediates the interaction. MCP1 interacts with the monocyte/microglia receptor CCR2. The interaction is critical to the function of MCP1 because CCR2(-/-) microglia do not undergo chemotaxis in response to MCP1 stimulation. We show that stimulation of microglia with FL-MCP1 or K104Stop-MCP1 triggers CCR2 internalization, whereas a mutant form unable to be cleaved at lysine 104 (K104A-MCP1) is relatively ineffective in this assay, suggesting that the C-terminal region interferes with the MCP1-CCR2 interaction. Moreover, FL-MCP1 and K104Stop-MCP1 stimulation leads to activation of Rac1, a small GTPase involved in cell migration. Conversely, MCP1-stimulated microglial migration is blocked by the Rac1 inhibitor, NSC23766, demonstrating the requirement for Rac1 effector pathways in this response. Taken together, we propose a model for MCP1 localization, activation, and function based on the initial presence and then removal of its C terminus, coupled with a requisite downstream signaling pathway from CCR2 stimulation to Rac1 activation.


[The effect of biogenic monoamine antagonists on the development of preimplantation mouse embryos cultured in vitro].
The effect of serotonin and adrenaline antagonists was tested on the early embryos of mice of three lines. All the substances tested produced an arrest or inhibition of cleavage division and the appearance of anomalies. Serotonin introduced in the incubation medium was effective against some serotoninolytics. We were unable to test the protective effect of adrenaline, as in the concentrations used it has its own effect on the development. From the data obtained, a conclusion is made of the existence in early mouse embryos of the structures sensitive to serotonin and adrenaline antagonists. The assumptions is made from the previously obtained data on the presence of biogenic monoamines in early mouse embryos, of functional activity of prospective mediators of the nervous system at the earliest stages of embryonic development of mammals.


A green and black tea extract benefits urological health in men with lower urinary tract symptoms.
The objective of this study was to examine the effects of a green and black tea extract blend [AssuriTEA Men's Health (AMH)] in men with lower urinary tract symptoms (LUTS). In this randomized, double-blind, placebo-controlled study, 46 men aged 30-70 with an American Urologic Association symptom score (AUAss) of at least 8 and up to 24 were randomized to 500 mg AMH, 1000 mg AMH, or placebo daily for 12 weeks. Measurements were taken at baseline (BL), week 6 and week 12 for AUAss, simple uroflowmetry, postvoid residual volume (PVR), C-reactive protein (CRP), Short-Form 36 Health Survey (SF-36), and International Index of Erectile Function (IIEF). A total of 40 subjects completed the study. AUAss decreased 34.5% from BL to week 12 in the 1000 mg AMH group (p = 0.008). At week 12, CRP increased in the 500 mg AMH (p = 0.003) and placebo (p = 0.012) groups from their BL levels but not in the 1000 mg group. Average urine flow (Qmean) increased in the 500 mg (p = 0.033) and 1000 mg AMH (p = 0.002) groups versus placebo. PVR decreased in the 1000 mg AMH group (p = 0.034) from BL at week 6. Treatment group effects were observed for the physical functioning and sexual desire domains of the SF-36 and IIEF (p = 0.051 and p = 0.005 respectively). AMH was well tolerated. Oral administration of AMH improved LUTS and quality of life in as little as 6 weeks.