C/EBPβ in bone marrow is essential for diet induced inflammation, cholesterol balance, and atherosclerosis.
Atherosclerosis is both a chronic inflammatory disease and a lipid metabolism disorder. C/EBPβ is well documented for its role in the development of hematopoietic cells and integration of lipid metabolism. However, C/EBPβ's role in atherosclerotic progression has not been examined. We assessed the impact of hematopoietic CEBPβ deletion in ApoE(-/-) mice on hyperlipidemia, inflammatory responses and lesion formation in the aorta. ApoE(-/-) mice were reconstituted with bone marrow cells derived from either WT or C/EBPβ(-/-) mice and placed on low fat or high fat/high cholesterol diet for 11 weeks. Hematopoietic C/EBPβ deletion in ApoE(-/-) mice reduced blood and hepatic lipids and gene expression of hepatic stearoyl CoA desaturase 1 and fatty acid synthase while expression of ATP binding cassette transporter G1, cholesterol 7-alpha-hydroxylase, and liver X receptor alpha genes were significantly increased. ApoE(-/-) mice reconstituted with C/EBPβ(-/-) bone marrow cells also significantly reduced blood cytokine levels and reduced lesion area in aortic sinuses compared with ApoE(-/-) mice reconstituted with WT bone marrow cells. Silencing of C/EBPβ in RAW264.7 macrophage cells prevented oxLDL-mediated foam cell formation and inflammatory cytokine secretion in conditioned medium. C/EBPβ in hematopoietic cells is crucial to regulate diet-induced inflammation, hyperlipidemia and atherosclerosis development.


Expression of fatty acid synthase in nonalcoholic fatty liver disease.
Nonalcoholic fatty liver disease (NAFLD) is characterized by hepatic lipid accumulation which starts with simple hepatic steatosis and may progress toward inflammation (nonalcoholic steatohepatitis [NASH]). Fatty acid synthase (FASN) catalyzes the last step in fatty acid biosynthesis, and thus, it is believed to be a major determinant of the maximal hepatic capacity to generate fatty acids by de novo lipogenesis. The aim of this study was to analyze the correlation between hepatic steatosis and inflammation with FASN expression. In vitro incubation of primary human hepatocytes with fatty acids dose-dependently induced cellular lipid-accumulation and FASN expression, while stimulation with TNF did not affect FASN levels. Further, hepatic FASN expression was significantly increased in vivo in a murine model of hepatic steatosis without significant inflammation but not in a murine NASH model as compared to control mice. Also, FASN expression was not increased in mice subjected to bile duct ligation, an experimental model characterized by severe hepatocellular damage and inflammation. Furthermore, FASN expression was analyzed in 102 human control or NAFLD livers applying tissue micro array technology and immunohistochemistry, and correlated significantly with the degree of hepatic steatosis, but not with inflammation or ballooning of hepatocytes. Quantification of FASN mRNA expression in human liver samples confirmed significantly higher FASN levels in hepatic steatosis but not in NASH, and expression of SREBP1, which is the main transcriptional regulator of FASN, paralleled FASN expression levels in human and experimental NAFLD. In conclusion, the transcriptional induction of FASN expression in hepatic steatosis is impaired in NASH, while hepatic inflammation in the absence of steatosis does not affect FASN expression, suggesting that FASN may serve as a new diagnostic marker or therapeutic target for the progression of NAFLD.


Cytotoxicity of murine B lymphocytes induced by human VH4-34 (VH4.21) gene-encoded monoclonal antibodies.
We have previously described specific binding and cytotoxicity of human B lymphocytes by VH4-34 gene-derived anti-i cold agglutinin (CA) mAbs. Here we demonstrate that the carbohydrate ligand recognized by human VH4-34 anti-i CA mAbs is also expressed on murine B lymphocytes. Similar to human B cells, binding of murine B lymphocytes by VH4-34-derived anti-i CA mAbs leads to rapid cytotoxicity of target cells as tested both in vitro and in vivo. Moreover, the mechanism leading to murine B cell death is also similar to human B cells, since morphologically identical membrane pores were detected within 15 min of mAb exposure by scanning electron microscopy. The conservation of the carbohydrate ligand across species provides an ideal system to study the function of human VH4-34 gene derived Abs in immune regulation.


A mobile phone-based care model for outpatient cardiac rehabilitation: the care assessment platform (CAP).
Cardiac rehabilitation programs offer effective means to prevent recurrence of a cardiac event, but poor uptake of current programs have been reported globally. Home based models are considered as a feasible alternative to avoid various barriers related to care centre based programs. This paper sets out the study design for a clinical trial seeking to test the hypothesis that these programs can be better and more efficiently supported with novel Information and Communication Technologies (ICT). We have integrated mobile phones and web services into a comprehensive home- based care model for outpatient cardiac rehabilitation. Mobile phones with a built-in accelerometer sensor are used to measure physical exercise and WellnessDiary software is used to collect information on patients' physiological risk factors and other health information. Video and teleconferencing are used for mentoring sessions aiming at behavioural modifications through goal setting. The mentors use web-portal to facilitate personal goal setting and to assess the progress of each patient in the program. Educational multimedia content are stored or transferred via messaging systems to the patients phone to be viewed on demand. We have designed a randomised controlled trial to compare the health outcomes and cost efficiency of the proposed model with a traditional community based rehabilitation program. The main outcome measure is adherence to physical exercise guidelines. The study will provide evidence on using mobile phones and web services for mentoring and self management in a home-based care model targeting sustainable behavioural modifications in cardiac rehabilitation patients. The trial has been registered in the Australian New Zealand Clinical Trials Registry (ANZCTR) with number ACTRN12609000251224.


Need for monitoring nonspecific bronchial hyperresponsiveness before and after isocyanate inhalation challenge.
Specific and nonspecific bronchial responsiveness may decline or disappear after cessation of exposure in the workplace in patients with occupational asthma, leading to false-negative specific inhalation challenge (SIC) results. Twenty-two patients with suspected diisocyanate-induced asthma were studied. SIC with diisocyanates (toluene diisocyanate [TDI] or hexamethylene diisocyanate [HDI]) was carried out in a 7-m(3) dynamic chamber up to a maximum concentration of 19 parts per billion for 120 min. Methacholine inhalation challenges were performed before and 24 h after SIC with TDI or HDI. Patients who did not show an asthmatic reaction after SIC but had a greater than twofold reduction in provocative concentration of methacholine causing a 20% fall in FEV(1) (PC(20)) after the first isocyanate challenge underwent a second isocyanate SIC 2 days later. The first SIC with isocyanates elicited an asthmatic reaction in 13 patients (59%). In five patients who did not show an asthmatic reaction after the first SIC, PC(20) exhibited more than a twofold reduction. In three of the five patients, a second SIC with isocyanates elicited an immediate positive asthmatic reaction. Therefore, 3 of 16 patients (19%) were ultimately shown to have bronchial responsiveness to isocyanate; occupational asthma was demonstrated due to post-SIC monitoring of bronchial hyperresponsiveness to methacholine. PC(20) should be systematically assessed before and after SIC with isocyanates in the absence of significant changes in FEV(1) during SIC to avoid false-negative results.


New directing groups for metal-catalyzed asymmetric carbon-carbon bond-forming processes: stereoconvergent alkyl-alkyl Suzuki cross-couplings of unactivated electrophiles.
The ability of two common protected forms of amines (carbamates and sulfonamides) to serve as directing groups in Ni-catalyzed Suzuki reactions has been exploited in the development of catalytic asymmetric methods for cross-coupling unactivated alkyl electrophiles. Racemic secondary bromides and chlorides undergo C-C bond formation in a stereoconvergent process in good ee at room temperature in the presence of a commercially available Ni complex and chiral ligand. Structure-enantioselectivity studies designed to elucidate the site of binding to Ni (the oxygen of the carbamate and of the sulfonamide) led to the discovery that sulfones also serve as useful directing groups for asymmetric Suzuki cross-couplings of racemic alkyl halides. To our knowledge, this investigation provides the first examples of the use of sulfonamides or sulfones as effective directing groups in metal-catalyzed asymmetric C-C bond-forming reactions. A mechanistic study established that transmetalation occurs with retention of stereochemistry and that the resulting Ni-C bond does not undergo homolysis in subsequent stages of the catalytic cycle.


The flavour of grape colour: anthocyanin content tunes aroma precursor composition by altering the berry microenvironment.
Anthocyaninless (white) instead of black/red (coloured) fruits develop in grapevine cultivars without functional VviMYBA1 and VviMYBA2 genes, and this conditions the colour of wines that can be produced. To evaluate whether this genetic variation has additional consequences on fruit ripening and composition, we performed comparisons of microenvironment, transcriptomics, and metabolomics of developing grapes between near-isogenic white- and black-berried somatic variants of Garnacha and Tempranillo cultivars. Berry temperature was as much as 3.5 ºC lower in white- compared to black-berried Tempranillo. An RNA-seq study combined with targeted and untargeted metabolomics revealed that ripening fruits of white-berried variants were characterized by the up-regulation of photosynthesis-related and other light-responsive genes and by their higher accumulation of specific terpene aroma precursors, fatty acid-derived aldehyde volatiles, and phenylpropanoid precursor amino acids. MYBA1-MYBA2 function proved essential for flavonol trihydroxylation in black-berried somatic variants, which were also characterized by enhanced expression of pathogen defence genes in the berry skin and increased accumulation of C6-derived alcohol and ester volatiles and γ-aminobutyric acid. Collectively, our results indicate that anthocyanin depletion has side-effects on grape composition by altering the internal microenvironment of the berry and the partitioning of the phenylpropanoid pathway. Our findings show how fruit colour can condition other fruit features, such as flavour potential and stress homeostasis.


Modulating Bioglass Concentration in 3D Printed Poly(propylene fumarate) Scaffolds for Post-Printing Functionalization with Bioactive Functional Groups.
Poly(propylene fumarate) (PPF) has shown potential for the treatment of bone defects as it can be 3D printed into scaffolds to suit patient-specific needs with strength comparable to that of bone. However, the lack of specific cell attachment and osteogenic signaling moieties have limited their utility as it is necessary to provide these signals to aid in bone tissue formation. To address this issue and provide a platform for functionalization, Bioglass (∼1-2 μm) microparticles have been incorporated into PPF to create a 3D printable resin with concentrations ranging from 0 to 10 wt %. The zero-shear viscosity of PPF-Bioglass resins increased proportionally from 0 to 2.5 wt % Bioglass, with values of 0.22 and 0.34 Pa·s, respectively. At higher Bioglass concentrations, 5 and 10 wt %, the resin viscosity increased to 0.44 and 1.31 Pa·s, exhibiting a 2- and 6-fold increase from the 0 wt % Bioglass resin. Despite this increase in viscosity, all resins remained printable with no print failures. In addition, the surface available Bioglass can tether catechol containing molecules for postprinting functionalization. Analysis of PPF-Bioglass functionalization using a catechol dye analyte shows functionalization increases with Bioglass concentration, up to 157 nmol/cm2, and demonstrates it is possible to modulate functionalization. This presents a versatile and highly translationally relevant strategy to functionalize 3D printed scaffolds post printing with a diverse array of functional species.


Coupling of voltage-dependent gating and Ba++ block in the high-conductance, Ca++-activated K+ channel.
Voltage-dependent Ca++-activated K+ channels from rat skeletal muscle were reconstituted into planar lipid bilayers, and the kinetics of block of single channels by Ba++ were studied. The Ba++ association rate varies linearly with the probability of the channel being open, while the dissociation rate follows a rectangular hyperbolic relationship with open-state probability. Ba ions can be occluded within the channel by closing the channel with a strongly hyperpolarizing voltage applied during a Ba++-blocked interval. Occluded Ba ions cannot dissociate from the blocking site until after the channel opens. The ability of the closed channel to occlude Ba++ is used as an assay to study the channel's gating equilibrium in the blocked state. The blocked channel opens and closes in a voltage-dependent process similar to that of the unblocked channel. The presence of a Ba ion destabilizes the closed state of the blocked channel, however, by 1.5 kcal/mol. The results confirm that Ba ions block this channel by binding in the K+-conduction pathway. They further show that the blocking site is inaccessible to Ba++ from both the cytoplasmic and external solutions when the channel is closed.


Systematic review and meta-analysis of linear and undulating periodized resistance training programs on muscular strength.
Periodization is known to improve training adaptations but the most effective periodization approach for muscular strength development for a wide variety of populations is yet to be determined. This systematic review and meta-analysis examined all studies directly comparing linear and undulating periodized resistance training programs to determine and compare their effects on muscular strength. A systematic search of the MEDLINE, SCOPUS, and SPORTDiscus databases revealed 17 studies satisfying the inclusion criteria. There were a total of 510 participants in the included studies. Sixteen studies reported significant increases in strength for both periodization approaches. Five studies reported significant differences in improvements between groups. The meta-analyses determined that there were no differences in the effectiveness of linear vs. undulating periodization on upper-body or lower-body strength. The short-term nature of studies and the previous training history of participants were identified as potential confounding factors in the interpretation of findings. The results suggest that novelty or training variety are important for stimulating further strength development. Few studies have examined the effect of periodization approaches in adolescent or athletic populations.


APF, HB-EGF, and EGF biomarkers in patients with ulcerative vs. non-ulcerative interstitial cystitis.
Interstitial cystitis (IC) is a chronic bladder disorder, with symptoms including pelvic and or perineal pain, urinary frequency, and urgency. The etiology of IC is unknown, but sensitive and specific biomarkers have been described, including antiproliferative factor (APF), heparin-binding epidermal growth factor-like growth factor (HB-EGF), and epidermal growth factor (EGF). However, the relative sensitivity of these biomarkers in ulcerative vs. nonulcerative IC is unknown, and these markers have yet to be validated in another laboratory. We therefore measured these markers in urine from patients with or without Hunner's ulcer, as well as normal controls, patients with bladder cancer, and patients with bacterial cystitis, at the First Hospital of China Medical University. Urine specimens were collected from two groups of Chinese IC patients (38 IC patients with Hunner's ulcers, 26 IC patients without Hunner's ulcers), 30 normal controls, 10 bacterial cystitis patients and 10 bladder cancer patients. APF activity was determined by measuring 3H-thymidine incorporation in vitro, and HB-EGF and EGF levels were determined by ELISA. APF activity (inhibition of thymidine incorporation) was significantly greater in all IC patient urine specimens than in normal control specimens or in specimens from patients with bacterial cystitis or bladder cancer (p < 0.0001 for each comparison). Urine HB-EGF levels were also significantly lower and EGF levels significantly higher in both groups of IC patients than in the three control groups (p < 0.0001 for each comparison). Although APF and HB-EGF levels were similar in ulcerative and nonulcerative IC patients, EGF levels were significantly higher in IC patients with vs. without ulcers (p < 0.004). These findings indicate that APF, HB-EGF and EGF are good biomarkers for both ulcerative and nonulcerative IC and validate their measurement as biomarkers for IC in Chinese patients.


Testing for altruism and social pressure in charitable giving.
Every year, 90% of Americans give money to charities. Is such generosity necessarily welfare enhancing for the giver? We present a theoretical framework that distinguishes two types of motivation: individuals like to give, for example, due to altruism or warm glow, and individuals would rather not give but dislike saying no, for example, due to social pressure. We design a door-to-door fund-raiser in which some households are informed about the exact time of solicitation with a flyer on their doorknobs. Thus, they can seek or avoid the fund-raiser. We find that the flyer reduces the share of households opening the door by 9% to 25% and, if the flyer allows checking a Do Not Disturb box, reduces giving by 28% to 42%. The latter decrease is concentrated among donations smaller than $10. These findings suggest that social pressure is an important determinant of door-to-door giving. Combining data from this and a complementary field experiment, we structurally estimate the model. The estimated social pressure cost of saying no to a solicitor is $3.80 for an in-state charity and $1.40 for an out-of-state charity. Our welfare calculations suggest that our door-to-door fund-raising campaigns on average lower the utility of the potential donors.


Combinatorial effects of double cardiomyopathy mutant alleles in rodent myocytes: a predictive cellular model of myofilament dysregulation in disease.
Inherited cardiomyopathy (CM) represents a diverse group of cardiac muscle diseases that present with a broad spectrum of symptoms ranging from benign to highly malignant. Contributing to this genetic complexity and clinical heterogeneity is the emergence of a cohort of patients that are double or compound heterozygotes who have inherited two different CM mutant alleles in the same or different sarcomeric gene. These patients typically have early disease onset with worse clinical outcomes. Little experimental attention has been directed towards elucidating the physiologic basis of double CM mutations at the cellular-molecular level. Here, dual gene transfer to isolated adult rat cardiac myocytes was used to determine the primary effects of co-expressing two different CM-linked mutant proteins on intact cardiac myocyte contractile physiology. Dual expression of two CM mutants, that alone moderately increase myofilament activation, tropomyosin mutant A63V and cardiac troponin mutant R146G, were shown to additively slow myocyte relaxation beyond either mutant studied in isolation. These results were qualitatively similar to a combination of moderate and strong activating CM mutant alleles alphaTmA63V and cTnI R193H, which approached a functional threshold. Interestingly, a combination of a CM myofilament deactivating mutant, troponin C G159D, together with an activating mutant, cTnIR193H, produced a hybrid phenotype that blunted the strong activating phenotype of cTnIR193H alone. This is evidence of neutralizing effects of activating/deactivating mutant alleles in combination. Taken together, this combinatorial mutant allele functional analysis lends molecular insight into disease severity and forms the foundation for a predictive model to deconstruct the myriad of possible CM double mutations in presenting patients.


Inhibition of cytotoxicity of lymphocytes by concanavalin A in vitro.
Human lymphocytes treated with the plant protein concanavalin A are stimulated to transform into blasts, without developing cytotoxicity for chicken erythrocytes. Prior treatment of lymphocytes with concanavalin A potentiated phytohemagglutinin-induced blast transformation and DNA synthesis but completely inhibited phytohemagglutinin-induced cytotoxicity. Inhibiton was not due to suppression of the mixed lymphocyte-erythrocyte aggregation normally caused by phytohemagglutinin. Inhibition of cytotoxicity was reversible when concanavalin A was removed from the lymphocytes by treatment with methyl-alpha-D-manno-pyranoside after 1 hour but not after 20 hours. The results indicate that blast transformation and cytotoxicity are separate expressions of lymphocyte stimulation.


Differential distribution of voltage-gated calcium channel alpha-2 delta (alpha2delta) subunit mRNA-containing cells in the rat central nervous system and the dorsal root ganglia.
Voltage-gated calcium channels (VGCCs) play an essential role in controlling neurotransmitter release, neuronal excitability, and gene expression in the nervous system. The distribution of cells that contain mRNAs encoding the auxiliary alpha2delta-1, alpha2delta-2, and alpha2delta-3 subunits of the VGCCs in the central nervous system (CNS) and the dorsal root ganglia (DRG) was examined in rats by using in situ hybridization. Specific labeling of alpha2delta-1, alpha2delta-2, and alpha2delta-3 mRNAs appeared to be largely confined to neurons and was widely, although differentially, distributed in the brain, the spinal cord, and the DRG. Importantly, alpha2delta-2 mRNA was found to be expressed in interneurons in the cortex, the hippocampus, the striatum, and in regions that contain dense cholinergic neurons. Our results suggest that different alpha2delta subunits may exert distinctive functions in the CNS. The alpha2delta-1 subunit mRNA is localized in brain regions known to be involved in cortical processing, learning and memory, defensive behavior, neuroendocrine secretion, autonomic activation, primary sensory transmission, and general arousal. The alpha2delta-2 subunit mRNA is present in brain regions known to modulate the overall activities of the cortex, the hippocampus, and the thalamus. The alpha2delta-2 subunit is also found in brain regions known to be involved in olfaction, somatic motor control, fluid homeostasis, ingestive and defensive behaviors, neuroendocrine functions, and circadian rhythm. In addition to being localized in brain regions that express alpha2delta-1 and alpha2delta-2 subunit mRNAs, alpha2delta-3 subunit mRNA is highly expressed in regions involved in auditory information processing and somatic movement.


metamedian: An R package for meta-analyzing studies reporting medians.
When performing an aggregate data meta-analysis of a continuous outcome, researchers often come across primary studies that report the sample median of the outcome. However, standard meta-analytic methods typically cannot be directly applied in this setting. In recent years, there has been substantial development in statistical methods to incorporate primary studies reporting sample medians in meta-analysis, yet there are currently no comprehensive software tools implementing these methods. In this paper, we present the metamedian R package, a freely available and open-source software tool for meta-analyzing primary studies that report sample medians. We summarize the main features of the software and illustrate its application through real data examples involving risk factors for a severe course of COVID-19.


[Axé, corporal practices, and AIDS in Africanist religions in Recife, Brazil].
This article analyzes the responses of Afro-Brazilian religions to the Aids epidemic in Recife, taking into consideration the religious symbolic structure. Drawing on participant observation and in-depth interviews conducted with Afro-Brazilian religious leaders and public health officials, it highlights the importance of "axé" _ the native category used to interpret corporal events _ in order to understand the history of Aids in this religious community. Axé is mystical energy, corporal vitality. It is manipulated in religious rituals and is symbolically associated with blood, perspiration and semen. In these times of HIV, the body scarification rituals and exchanges of fluids during sexual exchanges are the ways by which axé circulates among adept and are the core elements for its promotion, though they have also become a means for the transmission of the HIV virus. These elements were the basis of the dialogue between religious institutions and the public health system. This process generated changes in religious practices for regulation of social reproduction and the sexual life of the adepts.


Merging Clinical Cases, Client Communication, and Physiology to Enhance Student Engagement, Learning, and Skills.
Understanding disease processes, making diagnoses, and guiding clinical therapy are predicated on an understanding of normal physiologic function. However, we have observed that many first-year students fail to appreciate the important role that a clear understanding of normal function plays in becoming well-prepared, practicing veterinarians. Students also struggle with application of basic knowledge to the diagnosis and treatment of disease, as evidenced by poor performance on exam questions requiring application. The purpose of this project was to help students link the physiologic concepts in the classroom with clinical application, as well as to improve their ability to explain those concepts to a client. We found that, as a result of this assignment, students developed a deeper understanding of physiologic processes and their clinical relevance and, subsequently, felt more confident conveying this knowledge to simulated clients. Implementation of this case project has been very well received by the students. Students improved their grasp of the material, and they indicated that the project contributed positively to their motivation to study and learn physiology.


Sympathetic Skin Responses from the Neck Area in Patients with Unilateral Migraine.
In this study, in patients with unilateral migraine headache and in normal controls, it was aimed to assess the sympathetic function during attack, post attack, and interval periods and to compare these findings by recording sympathetic skin responses from the neck area, which was not studied before. A total of 37 unilateral patients with migraine (30 women, seven men) who fulfilled the criteria of International Headache Society (2004) were recruited from our outpatient clinic. The control group consisted of 21 healthy individuals (16 women, five men) who are employees or students of our Medical Faculty. Mean latency and maximum amplitude values of sympathetic skin responses obtained from neck areas of the patients during attack, post attack, and interval periods were calculated. We compared the mean latency and the maximum amplitude values of the symptomatic side with the data of the asymptomatic side and with the data of the control group. We also compared the responses of the patients with right-sided headache with the responses of the patients with left-sided headache. All statistical analyses were performed using SPSS. On the neck area, we observed sympathetic hypo-function in the attack and interval periods and a relative hyper-function in the post attack period bilaterally, regardless of the symptomatic side. These findings suggest that there is ongoing bilateral sympathetic hypo-function in the neck area and there occurs a temporary increase in the function of sympathetic sudomotor activity in the recovery period of headaches.


[Supervision of medical scientific research with mentally competent subjects is impossible without reviewing].
The object of reviewing medical scientific research in humans is to offer guarantees against, especially, violations of physical integrity and privacy. Since the scientific interest is a matter of first importance, the research has to be checked for its reasonableness and for the test subject's consent. Omission of these checks in case the risk or stress to be expected is slight is contrary to these criteria. Reversion of the sequence of informed consent and randomization is contrary to the requirement of consent. Free, informed consent may be assumed if in a trial with a control group the test subject is informed beforehand about the study design and the risks, but is not told if he is to receive a standard therapy, a placebo or a test medication.


Level of cardiovascular fitness and it's relationship with physical activity and body mass index in young adults of Islamabad.
To determine the level of cardiovascular fitness in young adults, and to find the relationship of maximum oxygen uptake with body mass index and physical activity. The cross-sectional study was conducted at two private universities in Islamabad, Pakistan, from January to June 2019, and comprised subjects of both genders aged 18-25 years with normal cardiopulmonary system. Physical activity was measured usint the short form of International Physical Activity Questionnaire, while maximum oxygen uptake was measured using the Bruce protocol. Data was analysed using SPSS 24. Of the 321 subjects, 59(18.4%) were males and 262(81.6%) were females. The overall mean age was 21.15±1.73 years. Overall, 152 (47.3%) subjects had normal body mass index, 86(26.8%) had high level of physical activity, and 85(26.5%) had moderate level. With respect to to maximum oxygen uptake, 28(47.5%) of the males were in the superior category, while 72(27.5%) women were in the good category. All the vitals along with dyspnoea and fatigue showed significant difference (p≤0.05) except oxygen saturation (p=0.062). Maximum oxygen uptake showed negative weak (p=0.40) correlation with body mass index and weak, positive and significant correlation (p=0.007) with physical activity. Majority of the young adults were found to be active as indicated by the levels of their maximum oxygen uptake and physical activity. Significant correlation was found between cardiovascular fitness and physical activity.


Are wait lists inevitable in subacute ambulatory and community health services? A qualitative analysis.
Objectives Wait lists are common in ambulatory and community-based services. The aim of the present study was to explore managers' perceptions of factors that contribute to wait times. Methods A qualitative study was conducted using semi-structured interviews with managers and team leaders of ambulatory and community health services within a large health network. Interviews were transcribed and coded, and the codes were then grouped into themes and subthemes. Results Representatives from 26 services participated in the project. Four major themes were identified. Three themes related to reasons and factors contributing to increased wait time for services (inefficient intake and scheduling processes; service disruptions due to human resource issues; and high service demand). A fourth theme related to staff attitudes towards wait times and acceptance and acknowledgement of wait lists. Conclusions Service providers perceive high demand to be a key driver of wait times, but a range of other factors also contributes and may represent opportunities for improving access to care. These other factors include improving process efficiencies, greater consistency of service delivery through more efficient management of human resources and shifting to more consumer-centred approaches in measuring wait times in order to drive improvements in patient flow. What is known about the topic? Wait times are common in out-patient and ambulatory services. These services experience high demand, which is likely to continue to grow as health service delivery shifts from hospital to community settings. What does this paper add? Although demand is an important driver of wait times, there are other modifiable factors that also contribute, including process inefficiencies and service disruption related to human resource issues. An underlying staff attitude of acceptance of wait times appears to be an additional barrier to improving access. What are the implications for practitioners? The findings of the present study suggest that there are opportunities for improving access to ambulatory and community health services through more efficient use of existing resources. However, a more consumer-focused approach regarding acceptability of wait times is needed to help drive change.


A randomized clinical trial to test the efficacy of a family-focused, culturally informed therapy for schizophrenia.
Research strongly suggests that family interventions can benefit patients with schizophrenia, yet current interventions often fail to consider the cultural context and spiritual practices that may make them more effective and relevant to ethnic minority populations. We have developed a family focused, culturally informed treatment for schizophrenia (CIT-S) patients and their caregivers to address this gap. Sixty-nine families were randomized to either 15 sessions of CIT-S or to a 3-session psychoeducation (PSY-ED) control condition. Forty-six families (66.7%) completed the study. The primary aim was to test whether CIT-S would outperform PSY-ED in reducing posttreatment symptom severity (controlling for baseline symptoms) on the Brief Psychiatric Rating Scale. Secondary analyses were conducted to test whether treatment efficacy would be moderated by ethnicity and whether patient-therapist ethnic match would relate to efficacy and patient satisfaction with treatment. Patients included 40 Hispanic/Latinos, 14 Whites, 11 Blacks, and 4 patients who identified as "other." In line with expectations, results from an ANCOVA indicated that patients assigned to the CIT-S condition had significantly less severe psychiatric symptoms at treatment termination than did patients assigned to the PSY-ED condition. Patient ethnicity and patient-therapist ethnic match (vs. mismatch) did not relate to treatment efficacy or satisfaction with the intervention. Results suggest that schizophrenia may respond to culturally informed psychosocial interventions. The treatment appears to work equally well for Whites and minorities alike. Follow-up research with a matched length control condition is needed. Further investigation is also needed to pinpoint specific mechanisms of change.


Using item response theory to model multiple phenotypes and their joint heritability in family data.
Many important complex diseases are composed of a series of phenotypes, which makes the disease diagnosis and its genetic dissection difficult. The standard procedures to determine heritability in such complex diseases are either applied for single phenotype analyses or to compare findings across phenotypes or multidimensional reduction procedures, such as principal components analysis using all phenotypes. However each method has its own problems and the challenges are even more complex for extended family data and categorical phenotypes. In this paper, we propose a methodology to determine a scale for complex outcomes involving multiple categorical phenotypes in extended pedigrees using item response theory (IRT) models that take all categorical phenotypes into account, allowing informative comparison among individuals. An advantage of the IRT framework is that a straightforward joint heritability parameter can be estimated for categorical phenotypes. Furthermore, our methodology allows many possible extensions such as the inclusion of covariates and multiple variance components. We use Markov Chain Monte Carlo algorithm for the parameter estimation and validate our method through simulated data. As an application we consider the metabolic syndrome as the multiple phenotype disease using data from the Baependi Heart Study consisting of 1,696 individuals in 95 families. We adjust IRT models without covariates and include age and age squared as covariates. The results showed that adjusting for covariates yields a higher joint heritability (ĥ2=0.53) than without co variates (ĥ2=0.21) indicating that the covariates absorbed some of the error variance.


Comparing Measurement Properties of the English EQ-5D-Y 3-Level Version With the 5-Level Version in South Africa.
This study aimed to compare the performance of the EQ-5D-Y 3-level version (Y-3L) and the expanded 5-level version, the EQ-5D-Y 5-level version (Y-5L). Children or adolescents with an acute or chronic health condition and from the general population completed the Y-5L and Y-3L self-report questionnaires. The performance of the Y-5L and Y-3L was determined by comparing feasibility, redistribution of dimension responses, discriminatory power, validity, and test-retest reliability. A total of 550 children/adolescents completed baseline measures and 173 completed repeat measures. The ceiling effect decreased by 15% from the Y-3L to Y-5L. Informativity of dimensions improved by 0.094 on the Y-5L. There was a range of 4% to 9% inconsistent responses moving from the Y-3L to Y-5L. Convergent validity of the visual analog scale and Y-3L and Y-5L dimensions was similar and weak to moderate (rs range 0.18-0.38) but similar and strong on paired Y-3L and Y-5L dimensions: Kendall tau B (range 0.69-0.80) and gamma (range 0.92-0.98). The Y-5L and Y-3L showed moderate to substantial agreement for test-retest reliability across dimensions and visual analog scale scores in stable chronic health conditions and fair agreement for the general population. The Y-5L is a valid, reliable extension of the Y-3L for children or adolescents across health conditions and healthy children/adolescents. The expanded levels reduced the ceiling effect. The relative informativity of report across dimensions increased on the Y-5L compared with the Y-3L with retention of the evenness of reporting. The convergent validity and test-retest reliability of the Y-5L was comparable with the Y-3L.


Harsh environments promote alloparental care across human societies.
Alloparental care is central to human life history, which integrates exceptionally short interbirth intervals and large birth size with an extended period of juvenile dependency and increased longevity. Formal models, previous comparative research, and palaeoanthropological evidence suggest that humans evolved higher levels of cooperative childcare in response to increasingly harsh environments. Although this hypothesis remains difficult to test directly, the relative importance of alloparental care varies across human societies, providing an opportunity to assess how local social and ecological factors influence the expression of this behaviour. We therefore, investigated associations between alloparental infant care and socioecology across 141 non-industrialized societies. We predicted increased alloparental care in harsher environments, due to the fitness benefits of cooperation in response to shared ecological challenges. We also predicted that starvation would decrease alloparental care, due to prohibitive energetic costs. Using Bayesian phylogenetic multilevel models, we tested these predictions while accounting for potential confounds as well as for population history. Consistent with our hypotheses, we found increased alloparental infant care in regions characterized by both reduced climate predictability and relatively lower average temperatures and precipitation. We also observed reduced alloparental care under conditions of high starvation. These results provide evidence of plasticity in human alloparenting in response to ecological contexts, comparable to previously observed patterns across avian and mammalian cooperative breeders. This suggests convergent social evolutionary processes may underlie both inter- and intraspecific variation in alloparental care.


Glucosensing in an immortalized adrenomedullary chromaffin cell line: role of ATP-sensitive K+ channels.
Using an immortalized adrenal chromaffin cell line (MAH cells), we investigated the cellular mechanisms underlying sensitivity to glucose-free solution (aglycemia) using ratiometric Ca2+ imaging and whole-cell recording. Though few cells (< 15%) responded to aglycemia with an increase in intracellular-free Ca2+ concentration ([Ca2+]i), in most cells (approximately 75%), aglycemia caused > 50% suppression of the Delta[Ca2+]i induced by the depolarizing stimulus, high (10 mM) K+. Moreover, in normal K+, the average aglycemia-induced rise in Cai2+ as well as the proportion of aglycemia-responsive cells increased in the presence of the K(ATP) channel blocker, glibenclamide. During membrane potential (Vm) measurements, aglycemia induced either hyperpolarization (6/20), depolarization (4/20) or no change in Vm. RT-PCR and Western blotting confirmed the presence of K(ATP) channel subunits Kir6.2 and SUR1 in MAH cells. These findings suggest a dual inhibitory and excitatory action of aglycemia in MAH cells, where activation of K(ATP) channels effectively inhibits or blunts the Delta[Ca2+]i due to the excitatory effect. Thus, this cell line appears as an attractive model for studying the molecular mechanisms of glucosensing.


[Hereditary neuropathy: variety of disease-causing genes and progress of molecular genetic diagnosis].
Inherited neuropathies are clinically and genetically heterogeneous. At least 30 genes have been associated with Charcot-Marie-Tooth disease (CMT) and related inherited neuropathies. Genetic studies have revealed that abnormalities in the following factors are the cause of inherited neuropathies: myelin components, transcription factors controlling myelination, myelin maintenance system, differentiation factors related to the peripheral nerve, neurofilaments, protein transfer system, mitochondrial proteins, DNA repair, RNA/protein synthesis, ion channels, and aminoacyl-tRNA synthetase. On the other hand, a precise molecular diagnosis is often needed to confirm a clinical diagnosis, offer genetic counseling to the patient and family, and provide prognostic information to the patient. Unfortunately, along with the increase in the number of genes that must be screened for mutations, the labor and reagent costs of molecular genetic testing have increased significantly. On the basis of the recent progress of DNA analysis methods, the use of resequencing microarray seems to be an economical and highly sensitive method to detect mutations. In this study, we attempted to screen for CMT patients mutations using these methods.


Percutaneous right portal vein embolization with polyvinyl alcohol particles in gastric cancer metastasis: report of a case.
Polyvinyl alcohol (PVA) particles are used for the embolization of various vascular tumors. They are also used before hepatic resection to embolize the ipsilateral portal vein, causing hypertrophy of the remaining liver. We report our first experience with portal vein embolization (PVE) with PVA particles to treat gastric cancer metastasis to the liver. PVE with PVA is a safe interventional radiologic procedure, which does not cause problems during surgery and can improve the outcome of hepatic resection.


[Characteristics of total blood protein measurement during infusion therapy].
The authors have modified R. Flores' method of protein measurements in solutions with bromophenol blue and suggest their modification for measurements of the blood serum total proteins. Experimental and clinical data have confirmed the significant effect of elevated blood levels of glucose and dextrans on the values of blood serum total proteins measured by the common biuret and refractometric methods. The bromophenol blue method is free from these disadvantages. The technique is recommended, first of all, for the infusion therapy departments. The method is highly specific, unsophisticated, and rapid, and is recommended for wide practical use in both inpatient and outpatient clinics. It can be used to measure proteins in other biologic fluids and solutions, provided its levels are not lower than 100 micrograms/ml.


[Effect of (2"R)-4'-O-tetrahydropyranyladriamycin, a new antitumor antibiotic, on the cardiac function of hamsters].
Cardiovascular effects of (2''R)-4'-O-tetrahydropylanyladriamycin X HCl (THP) and doxorubicin (adriamycin, ADM) were studied in hamsters. In experiments to observe acute effects, THP was administered intravenously at a dose of 12.5, 25.0 or 50.0 mg/kg, and ADM at 1.56, 3.13 or 6.25 mg/kg was given to different subjects. The THP caused slight ECG alterations at a dose of 12.5 mg/kg. At a dose of 25.0 mg/kg or 50.0 mg/kg, THP caused moderate to remarkable alterations in ECG like a widening of PR and PRc interval, A-V block, ST segment depression and T wave flattening. The ADM caused moderate to remarkable alterations in ECG at a dose of 3.13 mg/kg or 6.25 mg/kg, including arrhythmia, bradycardia, A-V block, ST segment changes and T wave flattening. These changes caused by THP and ADM recovered within 5 approximately 10 minutes after injection. Alterations in the ultrastructure of the myocardium caused by THP at a dose of 50.0 mg/kg included some cells with slight changes like swelling of mitochondria, focal intracellular edema, and enlargement of myofibrils. The ADM, at a dose of 3.13 mg/kg, induced severer swelling of mitochondria than THP, dilatation of sarcoplasmic reticulum, intracellular edema, and disorganization of myofilaments. At a dose of 6.25 mg/kg of ADM, these changes became more pronounced. In experiments to observe subacute effects, hamsters were treated with THP or ADM by daily intraperitoneal injections for 15 consecutive days, and then allowed to be recovered for 15 days. Dose levels of THP or ADM were 0.125, 0.25, 0.5 and 1.0 mg/kg. General toxicity, ECG, hematological and blood biochemical analysis, and electron microscopic examination were studied. In the ECG study, THP-treated hamsters showed a reversible elevation of R wave amplitude at a daily dose of 0.5 mg/kg. Widening of PR and PRc interval, elevation of R and S wave amplitude, and reduction of T wave amplitude were observed at a daily dose of 1.0 mg/kg of THP. Hamsters treated with ADM showed increase of heart rate, reduction of T wave amplitude, and shortening of PR and PRc interval at a daily dose of 0.5 mg/kg. Severe changes were observed at a daily dose of 1.0 mg/kg of ADM including an increase of heart rate, elevation of R wave amplitude, reduction of S and T wave amplitude, and shortening of QT interval. The electron microscopic examination revealed that THP-treated hamsters showed separation of intercalated discs, formation of myelin structure, and dilatation of T-tubules at a daily dose of 1.0 mg/kg. Similar changes were caused by ADM at a daily dose of 0.25 to 1.0 mg/kg.(ABSTRACT TRUNCATED AT 400 WORDS)


The transcriptional coactivator SAYP is a trithorax group signature subunit of the PBAP chromatin remodeling complex.
SWI/SNF ATP-dependent chromatin remodeling complexes (remodelers) perform critical functions in eukaryotic gene expression control. BAP and PBAP are the fly representatives of the two evolutionarily conserved major subclasses of SWI/SNF remodelers. Both complexes share seven core subunits, including the Brahma ATPase, but differ in a few signature subunits; POLYBROMO and BAP170 specify PBAP, whereas OSA defines BAP. Here, we show that the transcriptional coactivator and PHD finger protein SAYP is a novel PBAP subunit. Biochemical analysis established that SAYP is tightly associated with PBAP but absent from BAP. SAYP, POLYBROMO, and BAP170 display an intimately overlapping distribution on larval salivary gland polytene chromosomes. Genome-wide expression analysis revealed that SAYP is critical for PBAP-dependent transcription. SAYP is required for normal development and interacts genetically with core- and PBAP-selective subunits. Genetic analysis suggested that, like BAP, PBAP also counteracts Polycomb silencing. SAYP appears to be a key architectural component required for the integrity and association of the PBAP-specific module. We conclude that SAYP is a signature subunit that plays a major role in the functional specificity of the PBAP holoenzyme.


Analysis of the molecular composition of Ro ribonucleoprotein complexes. Identification of novel Y RNA-binding proteins.
Human Ro ribonucleoproteins (RNPs) are composed of one of the four small Y RNAs and at least two proteins, Ro60 and La; association of additional proteins including the Ro52 protein and calreticulin has been suggested, but clear-cut evidence is still lacking. Partial purification of Ro RNPs from HeLa S100 extracts allowed characterization of several subpopulations of Ro RNPs with estimated molecular masses of between 150 and 550 kDa. The majority of these complexes contained Ro60 and La, whereas only a small proportion of Ro52 appeared to be associated with Ro RNPs. To identify novel Y RNA-associated proteins in vitro, binding of cytoplasmic proteins to biotinylated Y RNAs was investigated. In these reconstitution experiments, several proteins with estimated molecular masses of 80, 68, 65, 62, 60 and 53 kDa, the latter two being immunologically distinct from Ro60 and Ro52, respectively, appeared to bind specifically to Y RNAs. Furthermore, autoantibodies to these proteins were found in sera from patients with systemic lupus erythematosus. The proteins bound preferentially to Y1 and Y3 RNA but, with the exception of the 53-kDa protein, only weakly to Y4 RNA and not at all to Y5 RNA. Coprecipitation of the 80, 68, 65, and 53-kDa proteins by antibodies to Ro60 and La was observed, suggesting that at least a proportion of the novel proteins may reside on the same particles as La and/or Ro60. Finally, the binding sites for these proteins on Y1 RNA were clearly distinct from the Ro60-binding site involving a portion of the large central loop 2, which was found to be indispensable for binding of the 80, 68, 65 and 53-kDa proteins, as well as the stem 3-loop 3 and stem 2-loop 1 regions. Interestingly, truncation of the La-binding site resulted in decreased binding of the novel proteins (but not of Ro60), indicating La to be required for efficient association. Taken together, these results suggest the existence of further subpopulations of Ro RNPs or Y RNPs, consistent with the heterogeneous characteristics observed for these particles in the biochemical fractionation experiments.


A mechanism for altered flexibility in human skeletal muscle.
1. We investigated the effect of a long-term stretching regimen on the tissue properties and stretch tolerance of human skeletal muscle. 2. Resistance to stretch was measured as torque (in N m) offered by the hamstring muscle group during passive knee extension while electromyographic (EMG) activity, knee joint angle and velocity were continuously monitored during a standardized stretch manoeuvre. Seven healthy subjects were tested before and after a 3 week training period using two separate protocols. Protocol 1 consisted of a slow stretch at 0.087 rad s-1 to a predetermined angle followed by a 90 s holding phase. Subjects were brought to the same angle before and after the training period. Protocol 2 was a similar stretch, but continued to the point of pain. 3. During protocol 1 the torque rose during the stretch and then declined during the holding phase. EMG activity was small and did not change significantly during the protocol. No significant differences in stiffness, energy and peak torque about the knee joint were seen as a result of the training. During protocol 2 the angle to which the knee could be extended was significantly increased as a result of the training. This was accompanied by a comparable increase in peak torque and energy. EMG activity was small and not affected by training. 4. It is concluded that reflex EMG activity does not limit the range of movement during slow stretches and that the increased range of motion achieved from training is a consequence of increased stretch tolerance on the part of the subject rather than a change in the mechanical or viscoelastic properties of the muscle.


When team members retaliate: The effect of LMX differentiation on team CWB.
Leader-member exchange (LMX) differentiation, or a leader's differentiated treatments among team members, is a prevalent phenomenon in any type of organization due to constraints of personal, social, and organizational resources. Despite numerous empirical examinations of such phenomenon, the impact of LMX differentiation on group outcomes is inconsistent and inconclusive. Based on team diversity framework, we approach LMX differentiation as disparity and draw on social comparison theory and social exchange theory to examine the positive effect of LMX differentiation on team counterproductive work behavior (CWB) through a supervisory justice climate. In addition, we investigate leader competence as an essential boundary condition that influences the impact of LMX differentiation on supervisory justice climate and subsequent team CWB. Using a sample of 94 teams in South Korea, we found that LMX differentiation negatively impacts supervisory justice climate, subsequently promoting team CWB. Furthermore, when team members perceive their leader as incompetent, the positive impact of LMX differentiation on team CWB is strengthened. Theoretical and practical implications and directions for future research are discussed.


Extracellular calmodulin-binding proteins in body fluids of animals.
The extracellular calmodulin-binding proteins (CaMBPs) were investigated in body fluids of animals by using the biotinylated calmodulin gel overlay method. Four major CaMBPs with molecular masses of 24, 31.5, 44/45 and 94 kDa were detected in serum, two of 24 and 63 kDa in bovine milk and three of 14, 24 and 52 kDa in human saliva. It suggested that extracellular CaMBPs exist commonly in body fluids of animals, and this result may provide a new clue for understanding the role of extracellular calmodulin.


Isolation of a 5.3-S calmodulin-deficient 3':5'-cyclic nucleotide phosphodiesterase from cardiac muscle.
1. In the presence of Ca2+, a 5.3-S 3':5'-cyclic nucleotide phosphodiesterase (EC 220.127.116.11) from bovine ventricle was isolated and purified by (NH4)2SO4 precipitation and DEAE-cellulose and Affi-Gel Blue chromatography. The enzyme activity was enriched 800-fold by these procedures. 2. Sucrose-density gradient centrifugation, gel filtration and non-denaturing polyacrylamide-gel electrophoresis resolved a single enzyme species with an Mr of 89 000. 3. Sodium dodecyl sulphate/polyacrylamide-gel electrophoresis of the purified enzyme demonstrated a prominent protein band at Mr 59000 and a minor band of Mr 28000. Calmodulin was not detected. 4. The hydrolysis of micromolar concentrations of 3':5'-cyclic guanosine monophosphate (cyclic GMP) but not 3':5'-cyclic adenosine monophosphate (cyclic AMP) was stimulated by calmodulin. 5. Anomalous biphasic kinetics plots were observed for both the catalysis of cyclic AMP and cyclic GMP hydrolysis. Kinetic plots became linear in the presence of calmodulin. 6. After several months of storage at -20 degrees C, the 5.3-S enzyme was transformed into a 6.2-S cyclic GMP-specific enzyme and a 4.4-S non-specific form.


Enzymatic characterization of the individual mammalian primase subunits reveals a biphasic mechanism for initiation of DNA replication.
The enzymatic mechanism of primase was investigated using Escherichia coli and baculoviral overexpressed mouse primase subunits, p49 and p58. Neither of the singly purified primase subunits displayed primase activity alone, but the p49 subunit was able to extend a riboprimer, indicating that this subunit contains an RNA polymerase activity. The p58 subunit cooperated with the p49 subunit in binding the initiating purine to form the initial dinucleotide. After initiation, the p49 subunit alone was sufficient to extend the growing primer, but both the rate of p49 primer extension and its stability were influenced by the p58 subunit. The Km(ATP) in primer synthesis on poly(dT) of the p49-p58 heterodimeric primase complex was 10-fold higher than the Km(ATP) of the single p49 subunit in a ribo(A) primer extension assay. In addition, labeled ATP cross-linked to both of the individually purified subunits but with a striking difference in affinities; cross-linking was 11-fold more efficient to the p49 subunit. The interaction of the two primase subunits with polymerase alpha was also investigated. Immunoprecipitation experiments indicate that only the p58 subunit directly contacts the p180 subunit of DNA polymerase alpha. Competition experiments in the coupled primase-polymerase assay with a catalytically inactive mutant of DNA polymerase alpha and the Klenow fragment suggest that the DNA polymerase alpha-primase complex does not dissociate from the primer during the transition from RNA to DNA synthesis.


Gene expression and alternative splicing contribute to adaptive divergence of ecotypes.
Regulation of gene expression is a critical link between genotype and phenotype explaining substantial heritable variation within species. However, we are only beginning to understand the ways that specific gene regulatory mechanisms contribute to adaptive divergence of populations. In plants, the post-transcriptional regulatory mechanism of alternative splicing (AS) plays an important role in both development and abiotic stress response, making it a compelling potential target of natural selection. AS allows organisms to generate multiple different transcripts/proteins from a single gene and thus may provide a source of evolutionary novelty. Here, we examine whether variation in alternative splicing and gene expression levels might contribute to adaptation and incipient speciation of dune-adapted prairie sunflowers in Great Sand Dunes National Park, Colorado, USA. We conducted a common garden experiment to assess transcriptomic variation among ecotypes and analyzed differential expression, differential splicing, and gene coexpression. We show that individual genes are strongly differentiated for both transcript level and alternative isoform proportions, even when grown in a common environment, and that gene coexpression networks are disrupted between ecotypes. Furthermore, we examined how genome-wide patterns of sequence divergence correspond to divergence in transcript levels and isoform proportions and find evidence for both cis and trans-regulation. Together, our results emphasize that alternative splicing has been an underappreciated mechanism providing source material for natural selection at short evolutionary time scales.


Dynamic ubiquitination determines transcriptional activity of the plant immune coactivator NPR1.
Activation of systemic acquired resistance in plants is associated with transcriptome reprogramming induced by the unstable coactivator NPR1. Immune-induced ubiquitination and proteasomal degradation of NPR1 are thought to facilitate continuous delivery of active NPR1 to target promoters, thereby maximising gene expression. Because of this potentially costly sacrificial process, we investigated if ubiquitination of NPR1 plays transcriptional roles prior to its proteasomal turnover. Here we show ubiquitination of NPR1 is a progressive event in which initial modification by a Cullin-RING E3 ligase promotes its chromatin association and expression of target genes. Only when polyubiquitination of NPR1 is enhanced by the E4 ligase, UBE4, it is targeted for proteasomal degradation. Conversely, ubiquitin ligase activities are opposed by UBP6/7, two proteasome-associated deubiquitinases that enhance NPR1 longevity. Thus, immune-induced transcriptome reprogramming requires sequential actions of E3 and E4 ligases balanced by opposing deubiquitinases that fine-tune activity of NPR1 without strict requirement for its sacrificial turnover.


Performance and amino acid utilization of early lactation dairy cows fed regular or reduced-fat dried distillers grains with solubles.
The objective of this study was to evaluate lactation response and AA utilization of early lactation cows fed 2 types of dried distillers grains with solubles (DG): regular (DDGS) or reduced-fat (RFDGS). Thirty-six Holstein cows 19.7+/-2.6 d in milk at the start of the experiment were used in a randomized complete block design for 14 wk including a 2-wk covariate period. Treatments consisted of the following diets: 1) control (CON) diet containing 0% DG; 2) diet containing 22% DDGS; and 3) diet containing 20% RFDGS. Distillers grains replaced soybean meal, expeller soybean meal, and soyhulls from the CON diet. Diets were formulated to be similar in crude protein, ether extract, neutral detergent fiber, and net energy for lactation concentrations. Dry matter intake (24.7 kg/d) and milk yield (39.3 kg/d) were similar for all diets. Milk fat and lactose percentages were unaffected by diets; however, protein percentage was greater for cows fed the DG diets compared with the CON diet. Consequently, milk protein yield was also greater for the DG diets compared with CON. Milk urea nitrogen decreased for cows fed DG diets and averaged 11.8, 10.9, and 10.1 mg/dL, respectively, for CON, DDGS, and RFDGS. Feed efficiency tended to be greater and N efficiency was greater for cows fed DG compared with CON. Body weight (711 kg), body weight change (+0.49 kg/d), and body condition score (3.36) were similar for all diets, but cows fed CON tended to gain more body condition (+0.14) than cows fed DG diets. Amino acid utilization was evaluated at the peak of milk production corresponding to wk 9 of lactation. Arterial Lys concentration was lower with DG diets (70.4, 58.6, and 55.8 microM/L). Cows fed DG had greater arterial Met concentration (21.3 microM) compared with CON (14.9 microM). Arterio-venous difference of Lys was similar across diets, whereas that of Met was greater for the DG diets compared with the CON diet (10.3 vs. 13.0 microM/L). Extraction efficiency of Lys by the mammary gland was greater for DG diets than for CON (76.1 vs. 65.4%). Mammary uptake of Lys (2.56 g/kg of milk) was similar for all diets, and the uptake of Met tended to increase in cows fed DG diets. Plasma glucose, triglyceride, and total cholesterol were unaffected by treatment; however, cows fed DG diets had lower beta-hydroxybutyrate and tended to have lower nonesterified fatty acid concentrations than cows fed the CON diet. Despite the apparent deficiency of Lys, milk protein percentage was increased in cows fed DG diets.


Effects of Exogenous Application of Protocatechuic Acid and Vanillic Acid to Chlorophylls, Phenolics and Antioxidant Enzymes of Rice (Oryza sativa L.) in Submergence.
In this study, effects from application of protocatechuic acid (PA) and vanillic acid (VA) and their mixture on the submergence tolerance of rice were examined. The treatment of 0.01 mM PA and VA did not show significant increase of rice growth as compared to the controls. However, at higher concentrations (0.1-1.0 mM), rice shoot was elevated in submergence by 20.8-22.4%. The survival percentage of rice seedlings at any dose of PA, VA and their mixture was significantly higher than the controls. In general, the mixture of PA and VA was more active to promote shoot elongation and survival in submergence than sole treatment of either PA or VA. The amount of chlorophyll b by PA was significantly increased, while no change in chlorophyll a content was observed. VA remarkably reduced malondialdehyde quantity at three days of submergence, while no significant difference among treatment was observed in PA, the mixture, and respective controls. The two phenolic acids promoted contents of phenolics and flavonoids in rice leaves and roots, however the quantities of endogenous PA and VA in rice were not markedly differed after PA and VA treated on roots of rice seedlings. The ascorbate peroxidase and superoxide dismutase activities were enhanced, while the expression of genes encoding antioxidant enzymes was favored. VA increased the expression level of ascorbate peroxidase genes in higher levels than PA and their mixture, while no significant difference was observed in the other genes including superoxide dismutase, catalase, glutathione reductase, and peroxidase. Findings of this study showed that PA and VA increased the submergence tolerance of rice by promoting the photosynthetic and anti-oxidative processes in rice seedlings. The treatment of PA and VA mixture on seedling roots was potent to promote the submergence tolerance in rice.


The outcome of surgically treated femur fractures associated with long-term bisphosphonate use.
Bisphosphonates (BPs) evolved as the mainstay for the treatment of osteoporosis, reducing the incidence of fractures. Recently several publications described the occurrence of low-energy subtrochanteric and femoral shaft fractures associated with long-term BP use. The aim of this study was to describe the outcome of surgically treated femur fractures associated with prolonged BP use. Fifteen patients suffering from 17 atypical femoral fragility fractures associated with long-term (>3 years) BP use were located. Data included fracture type, time of BP use, last bone mineral density DEXA scores for the femoral neck and spine, type of surgery, and the need for revision. Fourteen female patients and one male patient were identified. The median age was 73 years (range, 51-80 years). The mean BP use was 7.8 years (range, 4-13 years). Fourteen patients had low-energy traumatic femoral shaft (proximal and distal) or low subtrochanteric fractures. The mean lumbar spine (for 13 patients) bone mineral density T-score was -3.0, whereas mean femoral neck T-score was -1.8 with only three patients in the osteoporotic range.Fracture healing after the first procedure for patients treated with nails was 54%, with 46% of patients requiring revision surgery. These included nail dynamization, exchange nailing, and one revision to a blade plate. All of these eventually healed. BP-related fractures are a recently described phenomenon. Despite initial osteoporosis, the DEXA scan may appear outside the osteoporotic range for the femoral neck in these patients. In addition, a much higher failure rate with intramedullary nailing requiring revision surgery may occur with these patients.


Imaging readouts as biomarkers or surrogate parameters for the assessment of therapeutic interventions.
Surrogate markers and biomarkers based on imaging readouts providing predictive information on clinical outcome are of increasing importance in the preclinical and clinical evaluation of novel therapies. They are primarily used in studies designed to establish evidence that the therapeutic principle is valid in a representative patient population or in an individual. A critical step in the development of (imaging) surrogates is validation: correlation with established clinical endpoints must be demonstrated. Biomarkers must not fulfill such stringent validation criteria; however, they should provide insight into mechanistic aspects of the therapeutic intervention (proof-of-mechanism) or document therapy efficacy with prognostic quality with regard to the long-term clinical outcome (proof of concept). Currently used imaging biomarkers provide structural, physiological and metabolic information. Novel imaging approaches annotate structure with molecular signatures that are tightly linked to the pathophysiology or to the therapeutic principle. These cellular and molecular imaging methods yield information on drug biodistribution, receptor expression and occupancy, and/or intra- and intercellular signaling. The design of novel target-specific imaging probes is closely related to the development of the therapeutic agents and should be considered early in the discovery phase. Significant technical and regulatory hurdles have to be overcome to foster the use of imaging biomarkers for clinical drug evaluation.


Ultrastructural identification of protein bodies, cellular markers of human catecholamine neurons, in a temporal lobe ganglioglioma.
A temporal lobe ganglioglioma, surgically removed from an 8-year-old body, and a human brainstem at the level of locus coeruleus (LC) were processed for light microscopy (LM), with formalin fixation and paraffin embedding, and for electron microscopy (EM) with glutaraldehyde fixation, potassium permanganate postfixation, phosphotungstic acid-hematoxylin block-staining, and epoxy resin embedding. The paraffin sections were stained with toluidine blue O/rhodamine B and observed under epi-fluorescence. The thin sections for EM were viewed directly without further staining. The neuronal neoplastic cells of ganglioglioma and the neurons of LC are known to produce catecholamines. Both also contain spherical protein bodies (pb), cellular markers that identify catecholamine neurons in humans. The ultrastructural characteristics of the pb in LC were compared with those of the pb in neoplastic ganglion cells. These bodies had an identical ultrastructure, in both tissues, consisting of electron-lucent core surrounded by an electron-dense thin rim. The rhodamine B-stained sections also emphasized the identical morphology of the pb in ganglioglioma and LC. Based on the EM comparison, these brightly fluorescing spherical bodies are ideal markers for identifying in LM, the clusters of large neoplastic cells, representing neurons, which are the most important clue to the correct diagnosis of gangliogliomas.


Effect of pregnancy, postnatal growth, and gender on renal sulfate transport.
Serum sulfate concentrations are increased in infants, young children, and pregnant women, compared with adult values. The objective of this investigation was to examine the influences of age, gender, and pregnancy on renal sulfate transport using guinea pigs as an animal model. Membrane vesicles were isolated from the kidney cortex of male animals at four different ages, from male and female adult animals, and from pregnant and nonpregnant female animals. There were no significant differences in marker enzymes for the brush-border membrane (BBM) or basolateral membrane (BLM) among all groups examined. Uptake was determined by a rapid filtration method and membrane fluidity by measuring the steady-state fluorescence anisotropy of 1,6-diphenyl-1,3,5-hexatriene. The Vmax values for Na+ /sulfate co-transport in BBM were significantly increased with decreasing age, whereas the Km for this process was unchanged. The Vmax and Km for Na + /sulfate co-transport in BBM of pregnant animals were significantly higher than the values in the nonpregnant group. Bicarbonate-driven anion exchange of sulfate in BLM was not different among the different age groups. The Vmax for the bicarbonate/sulfate exchange process in BLM was not different between pregnant and nonpregnant groups; however, the Km for this process in BLM of pregnant animals was significantly greater than the value in nonpregnant animals. There were no gender-related differences in sulfate transport in BBM or BLM isolated from adult male and female animals. Renal BBM fluidity was increased with decreasing age and in pregnant animals, suggesting that altered membrane fluidity may represent one possible mechanism to explain the increased sodium/sulfate uptake in young and pregnant animals. The higher Vmax for Na+/sulfate co-transport in young and pregnant animals suggests that there is an increased density of co-transporter protein or an increase in the rate of movement of the carrier protein (i.e., turnover) once loaded with sodium and sulfate. This increased conservation of inorganic sulfate in young and pregnant guinea pigs may be related to the increased demand for sulfated substrates, such as sulfated glycosaminoglycans, during growth and development.


Bite Reconstruction in the Aesthetic Zone Using One-Piece Bicortical Screw Implants.
The aim of this article was to present the clinical application of a new, smooth surfaced one-piece bicortical screw implant with immediate loading protocol. An 18-year-old, healthy male patient with a history of total dislocation and replantation of teeth 11 and 21 in early childhood was admitted to the clinic. Teeth 11 and 21 were extracted, and two long one-piece implants were inserted at extraction sockets in one surgical session under local anesthesia. Temporary composite crowns were placed in the patient on the same day. After 3 months, the single-phase two-layer impression was made and the composite crowns were replaced with metal-ceramic crowns. After 12 months, satisfactory aesthetic and functional results were obtained.


Transesophageal echocardiography (TEE) vs. transthoracic echocardiography (TTE) in assessing cardio-vascular sources of emboli in patients with acute ischemic stroke.
Transesophageal echocardiography (TEE) is a valuable tool in the evaluation of ischemic stroke patients. However, the real clinical impact of additional TEE data remains to be defined. Our purpose was to evaluate the impact of TEE on the management of patients at low risk for cardiogenic embolism. We studied 68 patients (57+/-10 years old) with ischemic stroke at low-risk for cardiogenic embolism. Each patient underwent bilateral carotid ultrasound Doppler study, trans-thoracic echocardiography (TTE) and TEE in order to find out left atrial spontaneous echo contrast or intracavitary thrombi, communication or aneurysm of the inter-atrial septum, ventricular septal defect, patent foramen ovale, and the presence of intra-aortic atherosclerotic plaques or thrombi. In 28 out of 68 patients TEE found an abnormal lesion that has not been detected by TTE: there were 23 diffuse (>5mm) atherosclerotic atheromas in the aortic arch, 5 patent foramen ovale (PFO) lesions, 3 left atrial thrombi, 1 ventricular septal defect (VSD), and 1 atrial septal defect (ASD). 6 patients had more than 1 finding. These findings changed the management, and all 28 patients started to be treated with Coumadine instead of Aspirin. Patients with PFO were sent to close the shunt with a patch. In half of the patients TEE (but not TTE) found a significant lesion that changed our policy of management. None of these lesions were detected by TTE. It seems that TEE is mandatory in the evaluation of patients with acute ischemic stroke.


[Tuberculosis of the biliary system: presentation of a case and review of the literature].
We present the case of a female patient with tuberculosis of the gallbladder, the adipose tissue and lymph nodes surrounding the common bile duct; the patient presented with obstructive jaundice. We discuss the diagnostic criteria, approach to the biliary tract and treatment of this type of patients.


The Relation Between Patients' Beliefs About Pain Medication, Medication Adherence, and Treatment Outcome in Chronic Pain Patients: A Prospective Study.
Nonadherence to prescribed pain medication is common in chronic noncancer pain patients. Beliefs about pain medication have been reported to be associated with nonadherence behavior in cross-sectional studies. The aim of this study was to prospectively investigate the relationship between patients' beliefs about pain medication and their medication adherence and treatment outcome. Chronic noncancer pain patients completed a baseline questionnaire including the 47-item Pain Medication Attitudes Questionnaire, consisting of 7 subscales with regards to beliefs on prescribed medication. After 11 weeks, medication underuse and overuse were assessed by self-report. In addition, patient satisfaction with regards to the effect of prescribed pain medication and satisfaction as regards prescribed medication and care, and the burden of side effects were assessed. A total of 133 participants completed the baseline questionnaire, and 99 patients completed the follow-up questionnaire. Concerns over side effects at baseline were positively associated with underuse and the presence of side effects after 11 weeks. The perceived need was positively associated with overuse, whereas concerns over addiction were negatively associated with overuse. Concerns over tolerance were negatively associated with patient satisfaction with medication effects after 11 weeks. Concerns over tolerance and mistrust in the prescribing doctor were negatively associated with satisfaction about medication and care. Attitudes and concerns toward pain medication are associated with adherence patterns and outcome parameters. To improve medication adherence and therapy outcome, patient beliefs about pain medication should be taken into account by providing tailored education, adequate follow-up, or alternate therapy.


Globalisation in the time of COVID-19: repositioning Africa to meet the immediate and remote challenges.
The COVID-19 pandemic has ushered in a new climate of uncertainty which is fuelling protectionism and playing into nationalist narratives. Globalisation is under significant threat as governments scramble to reduce their vulnerability to the virus by limiting global trade and flows of people. With the imposition of border closures and strict migration measures, there have been major disruptions in Africa's global supply chains with adverse impacts on employment and poverty. The African economies overly reliant on single export-orientated industries, such as oil and gas, are expected to be severely hit. This situation is further aggravated by tumbling oil prices and a lowered global demand for African non-oil products. The agricultural sector, which should buffer these shocks, is also being affected by the enforcement of lockdowns which threaten people's livelihoods and food security. Lockdowns may not be the answer in Africa and the issue of public health pandemic response will need to be addressed by enacting context-specific policies which should be implemented in a humane way. In addressing the socioeconomic impact of COVID-19 on African nations, we argue that governments should prioritize social protection programmes to provide people with resources to maintain economic productivity while limiting job losses. International funders are committing assistance to Africa for this purpose, but generally as loans (adding to debt burdens) rather than as grants. G20 agreement so suspend debt payments for a year will help, but is insufficient to fiscal need. Maintaining cross-border trade and cooperation to continue generating public revenues is desirable. New strategies for diversifying African economies and limiting their dependence on external funding by promoting trade with a more regionalised (continental) focus as promoted by the African Continental Free Trade Agreement, while not without limitations, should be explored. While it is premature to judge the final economic and death toll of COVID-19, African leaders' response to the pandemic, and the support they receive from wealthier nations, will determine its eventual outcomes.


Treatment of canine hemangiosarcoma: 2000 and beyond.
Canine hemangiosarcoma (HSA) is an aggressive and malignant neoplasia with a grave prognosis. Surgery and chemotherapy have limited success in prolonging survival times and increasing quality of life in dogs with HSA. Advances in medical oncology are resulting in increased survival rates and a better quality of life for veterinary cancer patients. An understanding of mechanisms of metastasis has led to the development of new treatments designed to delay or inhibit tumor spread. Promising new treatment options include novel delivery systems (inhalation or intracavitary chemotherapy); use of immunomodulators such as liposome-encapsulated muramyl tripeptide-phosphatidylethanolamine; antimetastatic agents such as inhibitors of angiogenesis (interferons, thalidomide), matrix metalloproteinase inhibitors, and minocycline; dietary modifications; and gene therapy. Inhibitors of angiogenesis seem to be safe and, unlike conventional chemotherapy, do not induce drug resistance. Although many of the newer approaches are still under development and review, the use of multimodality therapy incorporating innovative treatment modalities may offer the best therapeutic option for dogs affected with HSA.


Microstructural white-matter abnormalities and their relationship with cognitive dysfunction in obsessive-compulsive disorder.
In recent years, diffusion tensor imaging (DTI) studies have detected subtle microstructural abnormalities of white matter (WM) in obsessive-compulsive disorder (OCD). However, findings have been inconsistent, and it is unclear whether WM abnormalities are related to cognitive processes. The aim of this study was to explore the relationship of WM alterations with cognitive variables in OCD in order to investigate the structural correlates of behaviorally relevant features of the disorder. We compared DTI-derived fractional anisotropy (FA), mean diffusivity (MD), and radial diffusivity (RD) measures between OCD patients (n = 16) and healthy controls (n = 18) using a whole-brain tract-based spatial statistics (TBSS) approach. We also explored the correlations of WM alterations with clinical and cognitive variables. Patients with OCD demonstrated increases in MD in the bilateral posterior corona radiata; left anterior corona radiata; bilateral superior longitudinal fasciculus; genu, body, and splenium of the corpus callosum; and left posterior limb of the internal capsule. An increase in RD values was also found in some of the same tracts (right posterior corona radiata, right superior longitudinal fasciculus, left anterior corona radiata, and corpus callosum). Furthermore, increased MD value in the internal capsule was correlated with the percentage of errors made during a target detection task, which was greater in the OCD group overall. These findings indicate that OCD patients show greater diffusivity in several white-matter regions. The correlation between cognitive performance and diffusivity in the internal capsule suggests that microstructural WM alternations may have functional consequences for the disorder.


Persistently low transplantation rate of ABO blood type O and highly sensitised patients despite alternative transplantation programs.
ABO blood type O and highly sensitised patients have the smallest chance to receive kidney transplantation. Do alternative donation programs increase this chance? In the period studied: 2323 patients were enlisted on the Rotterdam waiting list for a renal transplantation: 435 patients still waiting (WL), 464 delisted without transplantation (DWT). 1424 received deceased donor (DD, 535) or living donor (LD, 889, including 204 alternative) transplantation. Alternative LD programs in our centre are: paired kidney-exchange, altruistic with domino-paired donation and ABO-incompatible donation (ABOi). Compared to populations not transplanted, blood type O recipients are significantly underrepresented in DD and all LD transplantation populations, except the ABOi program. Highly sensitised patients are overrepresented in DD, but underrepresented in all LD transplantation populations. The high transplantation rate of highly sensitised patients was the result of Eurotransplant Acceptable mismatch program (AM). The LD ABOi and DD AM programs are the only alternative donation programs favourable for patients with low chances. While the contribution of direct LD transplantations will increase in time, the relative success rate of low-chance patients will decrease. Beside increasing LD ABOi transplantation, a new DD allocation model favouring both highly immunised and blood type O patients is essential.


Screening and Follow-Up Care for Perinatal Mood and Anxiety Disorders at a Rural Obstetric Clinic.
To improve screening for perinatal mood and anxiety disorders (PMAD) and follow-up care while balancing team workload. Four rapid plan-do-study-act cycles were implemented over 8 weeks. At baseline, only 2% of patients with PMAD were identified at a rural obstetric clinic, and none (n = 0 of 50) received screening with a validated tool. Of the 12 patients who had a current or prior history of PMAD, 92% (n = 11) were not screened for self-harm, and 67% (n = 8) received no referral. The clinic had no standardized care for PMAD. Patients (n = 253) screened at initial pregnancy intake, early in the third trimester, and at the 6-week postpartum visit. Following the screening, brief intervention, and referral to treatment model, patients were screened using the Edinburgh Postnatal Depression Scale, and brief intervention and referral to treatment were used with a point-of-care checklist. Data were collected three times weekly for run chart analysis, and team surveys measured workload. At the end of 8 weeks, effective screening for PMAD and follow-up care were achieved for 98% of patients and included screening, education, shared decision-making for management, referral, and clinic and phone follow-up to support mental health care uptake. Standardizing screening and follow-up care can increase identification of PMAD and increase uptake of mental health care. For sustainability, a decision aid can streamline patient-provider communication and reduce visit length.


Evolution of diadromy in fish: insights from a tropical genus (Kuhlia species).
Diadromous species undergo regular migration between fresh and marine waters. This behavior is found in many species, including fish, mollusks, and crustaceans, some of which are commercially valuable species. Several attempts to trace the evolution of this behavior have been made in Salmonidae and Galaxiidae, but ambiguous phylogenies and multiple character state changes prevented unequivocal conclusions. The Kuhliidae family consists of 12 fish species that inhabit tropical islands in the Indo-Pacific region. The species have marine, partially catadromous, or fully catadromous life histories (i.e., they migrate from rivers to the sea to reproduce). The evolution of migratory behavior was traced on a well-resolved phylogeny. Catadromous Kuhlia species were basal, and partially catadromous and marine species formed derived monophyletic groups. This is, to our knowledge, the first time that a clear origin and polarity for the diadromous character has been demonstrated. We propose that the relative lack of resources in tropical, inshore, marine habitats and the ephemeral and isolated nature of freshwater environments of tropical islands, combined with phenotypic plasticity of migratory traits, play key roles in driving the evolution of diadromy in the Kuhliidae and probably in other groups. This work is an important starting point to understand the role of diadromy in speciation and adaptation in unstable habitats.


Motor imagery for gait rehabilitation in post-stroke hemiparesis.
Reports have described the contribution of motor imagery (MI) practice for improving upper-extremity functions in patients with hemiparesis following stroke. The purpose of this case report is to describe the use of MI practice to attempt to improve walking in an individual with hemiparesis. A 69-year-old man with left hemiparesis received MI gait practice for 6 weeks. Intervention focused on task-oriented gait and on impairments of the affected lower limb. Preintervention, midterm, postintervention, and follow-up measurements of temporal-distance stride parameters and sagittal kinematics of the knee joint were taken. At 6 weeks postintervention, the patient had a 23% increase in gait speed and a 13% reduction in double-support time. An increase in range of motion of the knees also was observed. No changes in gait symmetry were noted. The outcomes suggest that MI may be useful for the enhancement of walking ability in patients following stroke. Because improvement was mainly in temporal-distance gait variables and knee movement, imagery practice probably should focus on its specific impairments during gait in order to affect the performance of the paretic lower extremity.


The lasting effect of intergenerational wealth transfers: Human capital, family formation, and wealth.
Recent evidence indicates that inheritances and other intergenerational wealth transfers have only a limited effect on wealth inequality and the intergenerational transmission of financial well-being. In this study, we explore the role that human capital and family formation play in mediating the relationship between receiving a transfer and building wealth. We examine how educational attainment and family formation determine whether or not households are able to convert inheritances into greater assets, facilitating improved wealth accumulation. Using data from the Panel Study for Income Dynamics (PSID), we examine how these factors moderate wealth accumulation trajectories following a bequest or inter vivos gift. Results reveal that educational attainment and marriage each facilitate wealth accumulation following a transfer. Our evidence suggests that cumulative advantage processes produce divergent wealth accumulation trajectories but these are situated in important turning points in the life course.


A Pyridyl-Furan Series Developed from the Open Global Health Library Block Red Blood Cell Invasion and Protein Trafficking in Plasmodium falciparum through Potential Inhibition of the Parasite's PI4KIIIB Enzyme.
With the resistance increasing to current antimalarial medicines, there is an urgent need to discover new drug targets and to develop new medicines against these targets. We therefore screened the Open Global Health Library of Merck KGaA, Darmstadt, Germany, of 250 compounds against the asexual blood stage of the deadliest malarial parasite Plasmodium falciparum, from which eight inhibitors with low micromolar potency were found. Due to its combined potencies against parasite growth and inhibition of red blood cell invasion, the pyridyl-furan compound OGHL250 was prioritized for further optimization. The potency of the series lead compound (WEHI-518) was improved 250-fold to low nanomolar levels against parasite blood-stage growth. Parasites selected for resistance to a related compound, MMV396797, were also resistant to WEHI-518 as well as KDU731, an inhibitor of the phosphatidylinositol kinase PfPI4KIIIB, suggesting that this kinase is the target of the pyridyl-furan series. Inhibition of PfPI4KIIIB blocks multiple stages of the parasite's life cycle and other potent inhibitors are currently under preclinical development. MMV396797-resistant parasites possess an E1316D mutation in PfPKI4IIIB that clusters with known resistance mutations of other inhibitors of the kinase. Building upon earlier studies that showed that PfPI4KIIIB inhibitors block the development of the invasive merozoite parasite stage, we show that members of the pyridyl-furan series also block invasion and/or the conversion of merozoites into ring-stage intracellular parasites through inhibition of protein secretion and export into red blood cells.


Minimally Invasive Endoscopic-Assisted Lateral Lumbar Interbody Fusion: Technical Report and Preliminary Results.
Lateral lumbar interbody fusion (LLIF) is a highly useful lumbar fusion surgical technique for degenerative spinal disease. However, many complications have already been reported. The purpose of this study is to report the concept, surgical technique, and clinical results of the first 70 consecutive cases treated with a safer and minimally invasive endoscopic-assisted LLIF (ELLIF). This retrospective study included 70 cases involving 106 segments in which ELLIF was used to treat degenerative spinal disease. We examined the clinical progress, complications and analyzed radiographic images. Regarding the fusion rate, 49 cases involving 72 segments whose follow-up period was more than 7 months were evaluated. The mean of preoperative Numerical Rating Scale (NRS) was 7.0 and postoperative NRS was 1.4. Postoperative NRS had a significant correlation with the number of fusion segments (p = 0.028). The mean of preoperative disc space height, foraminal height, sagittal rotation angle, whole lumbar lordosis and sagittal translation distance were 3.3 mm, 14.3 mm, 2.4°, 9.7°, and 3.2 mm, respectively. Postoperative values were 9.4 mm, 17.9 mm, -4.9°, 36.3°, and 0.7 mm. The fusion rate was 79.2%. Complications included, transient psoas muscle weakness 1, sensory disturbance in the thigh 2, retroperitoneal injury 1, postoperative ileus possibly involving a retroperitoneal injury 1, and cage migration 4. Using the ELLIF in the degenerative spinal disease, we obtained good radiological reduction and good clinical results. Our study confirms that ELLIF is safer and provides better results for degenerative spinal disease. However, the issue of cage migration remains to be resolved.


Topiramate Monotherapy for Civilian Posttraumatic Stress Disorder: A Controlled Pilot Study.
Objective: To assess the efficacy, safety, and tolerability of topiramate for the treatment of posttraumatic stress disorder (PTSD) in civilians. Methods: This 12-week double-blind, randomized, placebo-controlled study enrolled 72 outpatients (aged 19-64 years) with a DSM-IV-TR diagnosis of non-combat-related PTSD and a score ≥ 50 on the Clinician-Administered PTSD Scale (CAPS). The primary efficacy endpoint, percent change in total CAPS score, and secondary efficacy measures were assessed by analysis of covariance. Safety assessments included monitoring of vital signs, physical examinations, clinical laboratory parameters, electrocardiograms, and adverse events (AEs). The study was conducted from October 2001 to March 2004. Results: The intent-to-treat (ITT) population (N = 68; mean age = 35 years; 87% women; 74% White) showed greater percent reduction in total CAPS scores with topiramate versus placebo (39.5% vs 29.5%), but the difference was not statistically significant (P = .31). Similarly, higher reductions with topiramate versus placebo were seen in the CAPS subscale scores for symptoms of reexperiencing (43.6% vs 34.8%), avoidance/numbing (38.3% vs 30.6%), and hyperarousal (36.6% vs 21.4%). However, these differences were not statistically significant. Six patients in the topiramate arm had a final CAPS score < 20, whereas only 2 in the placebo arm achieved the result (P = .075). The median final topiramate daily dose was 100 mg/d (range, 25-400 mg/d), and mean ± SD treatment duration was 55 ± 32 days, showing the tolerability of the medication. In topiramate-treated patients, treatment-emergent AEs included paresthesia, headache, fatigue, and insomnia; treatment-limiting AEs included influenza-like symptoms, agitation, cognitive problems not otherwise specified, and somnolence. However, a higher rate of AE-related discontinuation was seen in the placebo group than in the treatment group (26% vs 18%). Conclusions: In this 12-week civilian PTSD study, topiramate improved the primary and secondary outcome measures at a higher rate than did placebo, but the difference did not reach statistical significance. Further adequately powered studies may be warranted. Trial Registration: Clinical Trials.gov identifier: NCT00208130. Prim Care Companion CNS Disord 2023;25(5):23m03555. Author affiliations are listed at the end of this article.


Star-shaped poly(oligoethylene glycol) copolymer-based gels: Thermo-responsive behaviour and bioapplicability for risedronate intranasal delivery.
The aim of this work was to obtain an intranasal delivery system with improved mechanical and mucoadhesive properties that could provide prolonged retention time for the delivery of risedronate (RS). For this, novel in situ forming gels comprising thermo-responsive star-shaped polymers, utilizing either polyethylene glycol methyl ether (PEGMA-ME 188, Mn 188) or polyethylene glycol ethyl ether (PEGMA-EE 246, Mn 246), with polyethylene glycol methyl ether (PEGMA-ME 475, Mn 475), were synthesized and characterized. RS was trapped in the selected gel-forming solutions at a concentration of 0.2% w/v. The pH, rheological properties, in vitro drug release, ex vivo permeation as well as mucoadhesion were also examined. MTT assays were conducted to verify nasal tolerability of the developed formulations. Initial in vivo studies were carried out to evaluate anti-osteoporotic activity in a glucocorticoid induced osteoporosis model in rats. The results showed successful development of thermo-sensitive formulations with favorable mechanical properties at 37 °C, which formed non-irritant, mucoadhesive porous networks, facilitating nasal RS delivery. Moreover, sustained release of RS, augmented permeability and marked anti-osteoporotic efficacy as compared to intranasal (IN) and intravenous (IV) RS solutions were realized. The combined results show that the in situ gels should have promising application as nasal drug delivery systems.