Pathways of glucose catabolism in procyclic Trypanosoma congolense.
Studies of respiration on glucose in procyclic Trypanosoma congolense in the presence of rotenone, antimycin, cyanide, salicylhydroxamic acid and malonate have indicated the presence of NADH dehydrogenase, cytochrome b-c1, cytochrome aa3, trypanosome alternate oxidase and NADH fumarate reductase/succinate dehydrogenase pathway that contributes electrons to coenzyme Q of the respiratory chain. The rotenone sensitive NADH dehydrogenase, the trypanosome alternate oxidase, and cytochrome aa3 accounted for 24.5 +/- 6.5, 36.2 +/- 4.2 and 54.1 +/- 5.5% respectively of the total respiration. Activities of lactate dehydrogenase, NAD(+)-linked malic enzyme and pyruvate kinase were less than 6 nanomoles/min/mg protein suggesting that they play a minor role in energy metabolism of the parasite. Phosphoenolpyruvate carboxykinase, pyruvate dehydrogenase, succinate dehydrogenase, NADP(+)-linked malic enzyme, NADH fumarate reductase, malate dehydrogenase, and alpha-ketoglutarate dehydrogenase and glycerol kinase on the other hand had specific activities greater than 60 nanomoles/min/mg protein. These enzyme activities could account for the production of pyruvate, acetate, succinate and glycerol. The results further show that the amount of glycerol produced was 35-48% of the combined total of pyruvate, acetate and succinate produced. It is apparent that some of the glycerol 3-phosphate produced in glycolysis in the presence of salicylhydroxamic acid is dephosphorylated to form glycerol while the rest is oxidised via cytochrome aa3 to form acetate, succinate and pyruvate.


No evidence for long-term facilitation after episodic hypoxia in spontaneously breathing, anesthetized rats.
Repeated electrical or hypoxic stimulation of peripheral chemoreceptors has been shown to cause a persistent poststimulus increase in respiratory motoneuron activity, termed long-term facilitation (LTF). LTF after episodic hypoxia has been demonstrated most consistently in anesthetized, vagotomized, paralyzed, artificially ventilated rats. Evidence for LTF in spontaneously breathing animals and humans after episodic hypoxia is equivocal and may have been influenced by the awake state of the subjects in these studies. The present study was designed to test the hypothesis that LTF is evoked in respiratory-related tongue muscle and inspiratory pump muscle activities after episodic hypoxia in 10 spontaneously breathing, anesthetized, vagotomized rats. The animals were exposed to three (5-min) episodes of isocapnic hypoxia, separated by 5 min of hyperoxia (50% inspired oxygen). Genioglossus, hyoglossus, and inspiratory intercostal EMG activities, along with respiratory-related tongue movements and esophageal pressure, were recorded before, during, and for 60 min after the end of episodic isocapnic hypoxia. We found no evidence for LTF in tongue muscle (genioglossus, hyoglossus) or inspiratory pump muscle (inspiratory intercostal) activities after episodic hypoxia. Rather, the primary poststimulus effect of episodic hypoxia was diminished respiratory frequency, which contributed to a reduction in ventilatory drive.


Expression and characterisation of the N-terminal fragment of the HsdS subunit of M.EcoR124I.
The type IC modification methyltransferase M.EcoR124I is a trimeric enzyme of 162 kDa consisting of two copies of the modification subunit, HsdM, and a single DNA specificity subunit, HsdS. Studies to date have been largely restricted to the HsdM subunit or the intact methyltransferase, since the HsdS subunit is insoluble when expressed independently of HsdM. Using PCR, we have cloned and expressed 13 fragments of the gene for the HsdS subunit, including the sequences encoding each of the variable and conserved domains and various combinations of these. Only two of these fragments were found to be soluble, a 8.6 kDa fragment (S11) comprising the central conserved domain and a 25 kDa N-terminal fragment (S3) containing the N-terminal variable domain and the central conserved domain. Analysis of the larger of these fragments by gel retardation shows that the protein binds DNA in the presence of HsdM at a subunit stoichiometry of 1:1. Gel filtration and CD spectroscopy indicate that the protein is monomeric and predominantly alpha-helical.


Cell-printed hierarchical scaffolds consisting of micro-sized polycaprolactone (PCL) and electrospun PCL nanofibers/cell-laden alginate struts for tissue regeneration.
Hierarchical scaffolds consisting of micro-sized struts with inter-layered nanofibers between the struts are mechanically stable and biologically superior to conventionally fabricated rapid-prototyped scaffolds and electrospun nanofibers. However, although the hierarchical scaffolds overcome various disadvantages of conventional scaffolds, there are still some limitations, such as low cell migration in the thickness direction and non-homogeneous cell proliferation. To overcome these deficiencies, a new hierarchical scaffold supplemented with osteoblast-like cell (MG63)-laden alginate struts is proposed. To control cell proliferation in the thickness direction of the scaffold, the density of interlayered nanofibers was manipulated using various electrospinning deposition times (2, 5, 10, and 20 s). Using the appropriate interlayered fiber density (electrospin deposition time = 10 s) and cell-laden alginate struts, we can obtain significantly homogeneous cell distribution in the hierarchical scaffold.


Intramolecular Catalysis of the Hydrolysis of N-Dimethylaminomaleamic Acid.
N-Dimethylaminomaleamic acid decomposes in aqueous solution by intramolecular reaction of the ciscarboxy group with the substituted carbamyl group. The reaction follows pseudo first-order kinetics and shows a dependence on pH. This compound retards plant growth probably because 1,1-dimethylhydrazine and 1,1-dimethylhydrazinium hydrogen maleate are products of its decomposition. Under similar conditions N-dimethylaminosuccinamic acid was stable.


Perceptual adaptation affects attractiveness of female bodies.
We investigated whether short durations (5 minutes) of exposure to distorted bodies can change subsequent perceptions of attractiveness and normality. Participants rated 110 female bodies, varying in width, on either their attractiveness or normality before and after exposure to either extremely narrow (-50% of original width in Experiments 1 and 2) or extremely wide bodies (+50% of original width in Experiment 1, and +70% of original width in Experiment 2). In both experiments, the most attractive and most normal looking bodies became significantly and substantially narrower after exposure to narrow bodies. The most normal looking body changed significantly after exposure to wide bodies, but the most attractive body did not. These results show that perceptions of body attractiveness can be influenced by experience, but that there is an asymmetry between the effects of exposure to narrow and wide bodies. We consider a possible mechanism for this unexpected asymmetry, as well as possible implications for the effects of media exposure on body-image. The most attractive body shape was consistently narrower than the most normal looking body shape. Substantial changes in what looked normal were accompanied by congruent changes in what looked attractive, suggesting that a normal or average body shape may function as a reference point against which body attractiveness is judged.


The inhibitory effect of quercetin on asymmetric dimethylarginine-induced apoptosis is mediated by the endoplasmic reticulum stress pathway in glomerular endothelial cells.
Asymmetric dimethylarginine (ADMA) is considered an independent mortality and cardiovascular risk factor in chronic kidney disease (CKD) patients, and contributes to the development of renal fibrosis. Quercetin (QC), a natural component of foods, protects against renal injury. Here, we explored the possible mechanisms that are responsible for ADMA-induced renal fibrosis and the protective effect of QC. We found that ADMA treatment activated the endoplasmic reticulum (ER) stress sensor proteins phosphorylated protein kinase RNA-activated-like ER kinase (PERK) and inositol requiring-1α (IRE1), which correspondingly induced C/EBP homologous protein (CHOP) expression and phosphorylated c-Jun N-terminal kinase (JNK) phosphorylation in glomerular endothelial cells (GEnCs). Following this, ADMA promoted ER stress-induced apoptosis and resulted in transforming growth factor β (TGF-β) expression in GEnCs. SP600125, an inhibitor of JNK, and CHOP siRNA protected against ADMA-induced cell apoptosis and TGF-β expression. QC prevented ADMA-induced PERK and IRE1 apoptotic ER stress pathway activation. Also, ADMA-induced GEnCs apoptosis and TGF-β expression was reduced by QC. Overexpression of CHOP blocked QC-mediated protection from apoptosis in ER stressed cells. Overall, these observations indicate that ADMA may induce GEnCs apoptosis and TGF-β expression by targeting the PERK-CHOP and IRE1-JNK pathway. In addition, drugs such as QC targeting ER stress may hold great promise for the development of novel therapies against ADMA-induced renal fibrosis.


Interactions between planar grafted neurofilament side-arms.
The side-arms of neurofilaments (NFs) have been proposed to be highly disordered, leading to an entropically and electrostatically based repulsion that modulates interfilament spacing. To characterize the behavior of two interacting polymer brushes in a system of this type, we performed molecular dynamics simulations of neurofilament brushes using a four bead reduced amino acid set coarse-grained model. In these simulations, we examined components of the neurofilament brush, NF-L, NF-M, and phosphorylated NF-H (NF-HP), individually. Each protein type was grafted to planar surfaces and simulations were performed for a range of separations of two apposed grafted surfaces. The calculated force-separation curves show the force increases as the reciprocal separation as predicted for polyelectrolyte brushes at high salt. All three systems can be overlapped on a single force-separation curve, which is not expected given the variation in amino acid sequence and charges on the polymers. Examination of structural properties shows scaling behavior in the average brush height, end-to-end distance, and the density interpenetration. Some of this scaling can be understood in terms of treating the NF proteins as effective polyelectrolytes, but some cannot suggesting a distinct polyampholyte behavior. Correlations are found between oppositely charged residues in opposite brushes. However, these correlations are weak in comparison to the strong correlations within each brush. In comparison with recent experimental data that observes condensed and expanded gel states, our results suggest that the condensed state structure involves significant interdigitation of the side-arms.


New Anti-Cancer Strategies in Testicular Germ Cell Tumors.
The most common solid malignancy of young men aged 20 to 34 years is testicular germ cell tumor. In addition, the incidence of these tumors has significantly increased throughout the last years. Testicular germ cell tumors are classified into seminoma and nonseminoma germ cell tumors, which take in yolk sac tumor, embryonal cell carcinoma, choriocarcinoma, and teratoma. There are noteworthy differences about therapy and prognosis of seminomas and nonseminoma germ cell tumors, even though both share characteristics of the primordial germ cells. The study is focused on different molecular mechanisms strongly involved in testicular germ cell line tumors underlying new strategies to treat this human neoplasia. Bibliographic data from peer-reviewed research, patent and clinical trial literature, and around eighty papers and patents have been included in this review. Our study reveals that several biomarkers are usefully utilized to discriminate among different histotypes. Moreover, we found new patents regarding testicular germ cell tumor treatments such as the expression of claudin 6, monoclonal antibody (Brentuximab Vedotin), immune checkpoint blockade (ICB) with the FDA-approved drugs pembrolizumab and nivolumab or the oncolytic virus Pelareorep, the combination of selective inhibitors of Aurora kinase. Finally, the pathogenesis of testicular germ cell tumor needs to be deeply understood so that it will improve data on stem cells, tumorigenesis and disease tumor management by more selective treatment.


A multicenter family practitioners' research on Chronic Obstructive Pulmonary Disease screening using the COPD Assessment Test.
Spirometry is known to be a gold standard for the diagnosis of chronic obstructive pulmonary disease (COPD). COPD Assessment Test (CAT) is an eight-item questionnaire currently in use to evaluate patients with COPD. In the present study, we aimed to evaluate if CAT is an adequate tool for screening COPD. In total, 600 persons aging ⩾40 years old were randomly selected from three different family practice units located in the city center. CAT was asked to the participants and a spirometry was used to assess pulmonary obstruction. Pulmonary obstruction was defined as forced expiratory volume in first second/forced vital capacity (FEV1/FVC)<70% and then COPD diagnosis was confirmed with the reversibility test. The relationship between CAT results and pulmonary function test values was evaluated. In this sampling, the prevalence of COPD was 4.2%. Reliability of the CAT in the study group was acceptable (Cronbach's α: 0.84). The CAT scores was significantly higher in patients with COPD (P<0.001). There was a significant negative correlation between CAT score and FEV1, FVC and FEV1/FVC ratio (r=-0.31, P<0.001; r=-0.26, P<0.001; r=0.18, P=0.001). Among smokers, phlegm was the predominating symptom (P=0.01). Sensitivity of CAT was 66.67% and its specificity was 75.15% to determine COPD. CAT is a reliable questionnaire and there is an apparent relationship between the total CAT scores and COPD. However, CAT's ability to screen COPD is limited since it may miss the symptom-free cases.


Neonatal Resuscitation Program and Pediatric Advanced Life Support.
The need for delivery resuscitation of the newborn cannot be predicted in most cases; therefore it is judicious to train all providers who may be involved in the delivery of newborns to follow guidelines developed to improve outcome, especially in the presence of transitional asphyxia. The Neonatal Resuscitation Program emphasizes basic steps of warming, drying, suctioning, and adequately ventilating the newborn. It also addresses current theories regarding resuscitation of the low birthweight newborns, infants with meconium aspiration, and medication use. The NRP applies to all acute-care hospitals that provide delivery services and those at which a respiratory therapist is likely to be present in the high-risk delivery or unanticipated delivery-room resuscitation. Outcomes have not been well documented and more clinical research is needed to identify which therapeutic strategies promote the best survival in this population. A topic that should be included in the NRP of the future is exogenous surfactant delivery. Respiratory distress syndrome has been a significant cause of death and morbidity in prematurely born neonates. Exogenous surfactant therapy has had a dramatic effect on the death rate of premature infants and on the incidence of respiratory distress syndrome. Current methods of surfactant administration demand that personnel proficient in management of the low birthweight newborn be present. As hospitals with all levels of nurseries continue to receive the prematurely delivered newborn and better methods to administer surfactant are discovered, the NRP could add information and a skills laboratory on surfactant administration. A trained cadre of health professionals who are proficient in the specific resuscitation skills required in pediatric patients can make a difference. The infant and child have different anatomy, physiology, and disease etiology that need to be emphasized and understood by the pediatric caregiver. The Pediatric Advanced Life Support course focuses on early recognition and treatment of respiratory failure and shock. The possibility of improving outcome with properly trained prehospital providers is a topic of interest. Intubation in the field can be successful and may contribute to a reduction in mortality, although complications of field intubation need to be reduced.


Developmentally regulated actions of alcohol on hippocampal glutamatergic transmission.
Ethanol exposure during fetal development is a leading cause of learning disabilities. Studies suggest that it alters learning and memory by permanently damaging the hippocampus. It is generally assumed that this is mediated, in part, via alterations in glutamatergic transmission. Although NMDA receptors are presumed to be the most sensitive targets of ethanol in immature neurons, this issue has not been explored in the developing hippocampus. We performed whole-cell patch-clamp recordings in hippocampal slices from neonatal rats. Unexpectedly, we found that acute ethanol (10-50 mM) exposure depresses inward currents elicited by local application of exogenous AMPA, but not NMDA, in CA3 pyramidal neurons. These findings revealed a direct effect of ethanol on postsynaptic AMPA receptors. Ethanol significantly decreased the amplitude of both AMPA and NMDA receptor-mediated EPSCs evoked by electrical stimulation. This effect was associated with an increase in the paired-pulse ratio and a decrease in the frequency of miniature EPSCs driven by depolarization of axonal terminals. These findings demonstrate that ethanol also acts at the presynaptic level. Omega-conotoxin-GVIA occluded the effect of ethanol on NMDA EPSCs, indicating that ethanol decreases glutamate release via inhibition of N-type voltage-gated Ca2+ channels. In more mature rats, ethanol did not affect the probability of glutamate release or postsynaptic AMPA receptor-mediated currents, but it did inhibit NMDA-mediated currents. We conclude that the mechanism by which ethanol inhibits glutamatergic transmission is age dependent and challenge the view that postsynaptic NMDA receptors are the primary targets of ethanol early in development.


Differential carbohydrate binding and cell surface glycosylation of human cancer cell lines.
Currently there is only a modest level knowledge of the glycosylation status of immortalised cell lines that are commonly used in cancer biology as well as their binding affinities to different glycan structures. Through use of glycan and lectin microarray technology, this study has endeavoured to define the different bindings of cell surface carbohydrate structures to glycan-binding lectins. The screening of breast cancer MDA-MB435 cells, cervical cancer HeLa cells and colon cancer Caco-2, HCT116 and HCT116-FM6 cells was conducted to determine their differential bindings to a variety of glycan and lectin structures printed on the array slides. An inverse relationship between the number of glycan structures recognised and the variety of cell surface glycosylation was observed. Of the cell lines tested, it was found that four bound to sialylated structures in initial screening. Secondary screening in the presence of a neuraminidase inhibitor (4-deoxy-4-guanidino-Neu5Ac2en) significantly reduced sialic acid binding. The array technology has proven to be useful in determining the glycosylation signatures of various cell-lines as well as their glycan binding preferences. The findings of this study provide the groundwork for further investigation into the numerous glycan-lectin interactions that are exhibited by immortalised cell lines.


Outcome after conservative management or intervention for unruptured brain arteriovenous malformations.
Whether conservative management is superior to interventional treatment for unruptured brain arteriovenous malformations (bAVMs) is uncertain because of the shortage of long-term comparative data. To compare the long-term outcomes of conservative management vs intervention for unruptured bAVM. Population-based inception cohort study of 204 residents of Scotland aged 16 years or older who were first diagnosed as having an unruptured bAVM during 1999-2003 or 2006-2010 and followed up prospectively for 12 years. Conservative management (no intervention) vs intervention (any endovascular embolization, neurosurgical excision, or stereotactic radiosurgery alone or in combination). Cox regression analyses, with multivariable adjustment for prognostic factors and baseline imbalances if hazards were proportional, to compare rates of the primary outcome (death or sustained morbidity of any cause by Oxford Handicap Scale [OHS] score ≥2 for ≥2 successive years [0 = no symptoms and 6 = death]) and the secondary outcome (nonfatal symptomatic stroke or death due to bAVM, associated arterial aneurysm, or intervention). Of 204 patients, 103 underwent intervention. Those who underwent intervention were younger, more likely to have presented with seizure, and less likely to have large bAVMs than patients managed conservatively. During a median follow-up of 6.9 years (94% completeness), the rate of progression to the primary outcome was lower with conservative management during the first 4 years of follow-up (36 vs 39 events; 9.5 vs 9.8 per 100 person-years; adjusted hazard ratio, 0.59; 95% CI, 0.35-0.99), but rates were similar thereafter. The rate of the secondary outcome was lower with conservative management during 12 years of follow-up (14 vs 38 events; 1.6 vs 3.3 per 100 person-years; adjusted hazard ratio, 0.37; 95% CI, 0.19-0.72). Among patients aged 16 years or older diagnosed as having unruptured bAVM, use of conservative management compared with intervention was associated with better clinical outcomes for up to 12 years. Longer follow-up is required to understand whether this association persists.


Validity and reliability testing: Urdu translated modified Response to Symptoms Questionnaire.
To determine validity and reliability of the Urdu translated, modified "Response to symptoms questionnaire" (RSQ) among acute coronary syndrome (ACS) patients in Karachi. A qualitative, tool validation study. Two tertiary care hospitals in Karachi, the Aga Khan University Hospital, Karachi and the Karachi Institute of Heart Diseases, from December 2010 to April 2011. After making certain modifications, the original tool in English was translated into Urdu. Next, five cardiology experts evaluated the tool for its content and face validity. Test retest and inter rater reliabilities were computed for the RSQ using 5% of the total sample size of the parent study. Sufficient conceptual and semantic equivalence was found between the Urdu and English versions of the modified RSQ. Content validity index was calculated to be 1 for both relevance and linguistic clarity. Test retest and inter rater reliabilities were calculated to be 95.9% and K = 0.97, respectively. The Urdu translated modified RSQ has sufficiently acceptable content validity, test retest and inter rater reliability; hence, it should be used by the researchers for the evaluation of factors associated with pre-hospital delay among Urdu speaking ACS patient populations.


Task-specific enhancement of hippocampus-dependent learning in mice deficient in monoacylglycerol lipase, the major hydrolyzing enzyme of the endocannabinoid 2-arachidonoylglycerol.
Growing evidence indicates that the endocannabinoid system is important for the acquisition and/or extinction of learning and memory. However, it is unclear which endocannabinoid(s) play(s) a crucial role in these cognitive functions, especially memory extinction. To elucidate the physiological role of 2-arachidonoylglycerol (2-AG), a major endocannabinoid, in behavioral and cognitive functions, we conducted a comprehensive behavioral test battery in knockout (KO) mice deficient in monoacylglycerol lipase (MGL), the major hydrolyzing enzyme of 2-AG. We found age-dependent increases in spontaneous physical activity (SPA) in MGL KO mice. Next, we tested the MGL KO mice using 5 hippocampus-dependent learning paradigms (i.e., Morris water maze (MWM), contextual fear conditioning, novel object recognition test, trace eyeblink conditioning, and water-finding test). In the MWM, MGL KO mice showed normal acquisition of reference memory, but exhibited significantly faster extinction of the learned behavior. Moreover, they showed faster memory acquisition on the reversal-learning task of the MWM. In contrast, in the contextual fear conditioning, MGL KO mice tended to show slower memory extinction. In the novel object recognition and water-finding tests, MGL KO mice exhibited enhanced memory acquisition. Trace eyeblink conditioning was not altered in MGL KO mice throughout the acquisition and extinction phases. These results indicate that 2-AG signaling is important for hippocampus-dependent learning and memory, but its contribution is highly task-dependent.


Problematic social networking site usage and substance use by young adolescents.
Substance use and abuse by young adolescents has become a serious issue for public health services, and several socio-environmental factors can influence how vulnerable a young adolescent may be to their appeal. The present study was devised to examine whether substance use in early adolescence is associated with problematic social networking site usage (PSNSU). In the academic year 2013-2014, secondary schools in Padua (north-eastern Italy) were involved in a survey called "Pinocchio". A sample of 1325 pupils attending years 6 to 8 (i.e. aged from 11 to 13 years) completed self-administered questionnaires, in which PSNSU was measured by applying the DSM-IV criteria of dependence to identify any social network addiction disorder and its fallout on daily life. Multivariate analysis (ordered logistic regression) was performed to assess an adjusted association between young adolescents' substance use and PSNSU. The percentage of pupils classified as problematic social networking site users rose with age (from 14.6% in year 6 to 24.3% in year 7, and 37.2% in year 8), and it was higher in girls (27.1%) than in boys (23.6%). In a fully-adjusted model, PSNSU conferred a higher likelihood of being substance users (OR 2.93 95% CI 1.77-4.85). This study identified an association between PSNSU and the likelihood of substance use (smoking, alcohol and energy drink consumption), providing further evidence of the need to pay more attention to PSNSU in early adolescence.


Spin entanglement in supramolecular structures.
Molecular spin clusters are mesoscopic systems whose structural and physical features can be tailored at the synthetic level. Besides, their quantum behavior is directly accessible in the laboratory and their magnetic properties can be rationalized in terms of microscopic spin models. Thus they represent an ideal playground within solid state systems to test concepts in quantum mechanics. One intriguing challenge is to control entanglement between molecular spins. Here we show how this goal can be pursued by discussing specific examples and referring to recent achievements.


A Scoping Review of Digital Gaming Research Involving Older Adults Aged 85 and Older.
Interest in the use of digital game technologies by older adults is growing across disciplines from health and gerontology to computer science and game studies. The objective of this scoping review was to examine research evidence involving the oldest old (persons 85 years of age or greater) and digital game technology. PubMed, CINHAL, and Scopus were searched, and 46 articles were included in this review. Results highlighted that 60 percent of articles were published in gerontological journals, whereas only 8.7 percent were published in computer science journals. No studies focused directly on the oldest old population. Few studies included sample sizes greater than 100 participants. Seven primary and 34 secondary themes were identified, of which Hardware Technology and Assessment were the most common. Existing evidence demonstrates the paucity of studies engaging older adults 85 years of age and above regarding the use of digital gaming and highlights a new understudied cohort for further research focus. Recommendations for future research include intentional recruitment and proportionate representation of participants ≥85 years of age, large sample sizes, and explicit mention of specific numbers of participants ≥85 years of age, which are necessary to advance knowledge in this area. Integrating a rigorous and robust mixed-methods approach including theoretical perspectives would lend itself to further in-depth understanding and knowledge generation in this field.


Nucleus accumbens medium spiny neuron subtypes mediate depression-related outcomes to social defeat stress.
The nucleus accumbens is a critical mediator of depression-related outcomes to social defeat stress. Previous studies demonstrate distinct neuroplasticity adaptations in the two medium spiny neuron (MSN) subtypes, those enriched in dopamine receptor D1 versus dopamine receptor D2, in reward and reinforcement leading to opposing roles for these MSNs in these behaviors. However, the distinct roles of nucleus accumbens MSN subtypes, in depression, remain poorly understood. Using whole-cell patch clamp electrophysiology, we examined excitatory input to MSN subtypes and intrinsic excitability measures in D1-green fluorescent protein and D2-green fluorescent protein bacterial artificial chromosome transgenic mice that underwent chronic social defeat stress (CSDS). Optogenetic and pharmacogenetic approaches were used to bidirectionally alter firing of D1-MSNs or D2-MSNs after CSDS or before a subthreshold social defeat stress in D1-Cre or D2-Cre bacterial artificial chromosome transgenic mice. We demonstrate that the frequency of excitatory synaptic input is decreased in D1-MSNs and increased in D2-MSNs in mice displaying depression-like behaviors after CSDS. Enhancing activity in D1-MSNs results in resilient behavioral outcomes, while inhibition of these MSNs induces depression-like outcomes after CSDS. Bidirectional modulation of D2-MSNs does not alter behavioral responses to CSDS; however, repeated activation of D2-MSNs in stress naïve mice induces social avoidance following subthreshold social defeat stress. Our studies uncover novel functions of MSN subtypes in depression-like outcomes. Notably, bidirectional alteration of D1-MSN activity promotes opposite behavioral outcomes to chronic social stress. Therefore, targeting D1-MSN activity may provide novel treatment strategies for depression or other affective disorders.


A Moment Matching Approach for Generating Synthetic Data.
Synthetic data are becoming increasingly important mechanisms for sharing data among collaborators and with the public. Multiple methods for the generation of synthetic data have been proposed, but many have short comings with respect to maintaining the statistical properties of the original data. We propose a new method for fully synthetic data generation that leverages linear and integer mathematical programming models in order to match the moments of the original data in the synthetic data. This method has no inherent disclosure risk and does not require parametric or distributional assumptions. We demonstrate this methodology using the Framingham Heart Study. Existing synthetic data methods that use chained equations were compared with our approach. We fit Cox proportional hazards, logistic regression, and nonparametric models to synthetic data and compared with models fitted to the original data. True coverage, the proportion of synthetic data parameter confidence intervals that include the original data's parameter estimate, was 100% for parametric models when up to four moments were matched, and consistently outperformed the chained equations approach. The area under the curve and accuracy of the nonparametric models trained on synthetic data marginally differed when tested on the full original data. Models were also trained on synthetic data and a partition of original data and were tested on a held-out portion of original data. Fourth-order moment matched synthetic data outperformed others with respect to fitted parametric models but did not always outperform other methods with fitted nonparametric models. No single synthetic data method consistently outperformed others when assessing the performance of nonparametric models. The performance of fourth-order moment matched synthetic data in fitting parametric models suggests its use in these cases. Our empirical results also suggest that the performance of synthetic data generation techniques, including the moment matching approach, is less stable for use with nonparametric models. The benefits of the moment matching approach should be weighed against additional computational costs. In summary, our results demonstrate that the introduced moment matching approach may be considered as an alternative to existing synthetic data generation methods.


Methylmercury augments Nrf2 activity by downregulation of the Src family kinase Fyn.
Methylmercury (MeHg) is a potent developmental neurotoxicant that induces an oxidative stress response in the brain. It has been demonstrated that MeHg exposure increases nuclear factor erythroid 2-related factor 2 (Nrf2) activity. Nrf2 is a transcription factor that translocates to the nucleus in response to oxidative stress, and upregulates phase II detoxifying enzymes. Although, Nrf2 activity is augmented subsequent to MeHg exposure, it has yet to be established whether Nrf2 moves into the nucleus as a result. Furthermore, the potential effect MeHg might have on the non-receptor tyrosine kinase, Fyn, has not been addressed. Fyn phosphorylates Nrf2 in the nucleus, resulting in its inactivation, and consequent downregulation of the oxidative stress response. Here, we observe Nrf2 translocates to the nucleus subsequent to MeHg-induced oxidative stress. This response is concomitant with reduced Fyn expression and nuclear localization. Moreover, we detected an increase in phosphorylated Akt and glycogen synthase kinase 3 beta (GSK-3β) at activating and inhibitory sites, respectively. Akt phosphorylates and inhibits GSK-3β, which subsequently prevents Fyn phosphorylation to signal nuclear import. Our results demonstrate MeHg downregulates Fyn to maintain Nrf2 activity, and further illuminate a potential mechanism by which MeHg elicits neurotoxicity.


Cephalic phase of gastroduodenal alkaline secretion.
Alkaline secretion measured under basal conditions in the intact stomach of conscious dogs averaged 47 mumol/30 min and was about twice lower than that recorded in the proximal (approximately 7 cm long) portion of the duodenum. Vagal excitation elicited by sham feeding and insulin resulted in a marked stimulation of alkaline secretion both from the stomach and the duodenum. Atropine significantly reduced gastric and duodenal alkaline secretion under basal state. It abolished gastric and diminished duodenal alkaline response to sham feeding and insulin hypoglycemia, while propranolol was without significant influence. Indomethacin reduced by approximately 75% basal duodenal alkaline secretion but did not prevent the increment in alkaline response to vagal stimulation. We postulate the existence of the cephalic phase of gastroduodenal alkaline secretion, which seems to be cholinergically dependent in the stomach and partly of noncholinergic and nonadrenergic character but prostaglandin dependent in the duodenum.


Blinded sample size recalculation for clinical trials with normal data and baseline adjusted analysis.
Baseline adjusted analyses are commonly encountered in practice, and regulatory guidelines endorse this practice. Sample size calculations for this kind of analyses require knowledge of the magnitude of nuisance parameters that are usually not given when the results of clinical trials are reported in the literature. It is therefore quite natural to start with a preliminary calculated sample size based on the sparse information available in the planning phase and to re-estimate the value of the nuisance parameters (and with it the sample size) when a portion of the planned number of patients have completed the study. We investigate the characteristics of this internal pilot study design when an analysis of covariance with normally distributed outcome and one random covariate is applied. For this purpose we first assess the accuracy of four approximate sample size formulae within the fixed sample size design. Then the performance of the recalculation procedure with respect to its actual Type I error rate and power characteristics is examined. The results of simulation studies show that this approach has favorable properties with respect to the Type I error rate and power. Together with its simplicity, these features should make it attractive for practical application.


The cannabinoid receptor 1 and its role in influencing peripheral metabolism.
Evidence from in vitro and in vivo studies has demonstrated the deleterious pathological effects of a dysregulated endocannabinoid system. Increased stimulation of the cannabinoid receptor 1 (CB1 ) and subsequent downstream cellular signalling are both causative in the deleterious pathological effects observed in a number of diseases. When the CB1 cell signalling cascade is blocked, this results in whole body weight-loss, leading to a reduction in obesity and associated co-morbidities. In the central nervous system; however, CB1 antagonism results in adverse psychological side effects. Blockade of CB1 via peripheral acting compounds that do not cross the blood-brain barrier have been determined to have beneficial effects in metabolic tissues such as the liver and skeletal muscle. These results support the notion that peripheral blockade of CB1 using pharmacological antagonists is a viable target for the treatment of the current epidemic of obesity and its associated co-morbidities.


Late-Onset Kidney Biopsy-Associated Retroperitoneal Hemorrhage in Lupus Nephritis: A Case Report of Pseudoaneurysm and Microaneurysm Formations.
Late-onset retroperitoneal hemorrhage from renal intraparenchymal pseudoaneurysm (RIP) following a kidney biopsy is an extremely rare complication but should not be ignored, especially in high-risk populations. Here, we introduce a 32-year-old Caucasian female who presented with sudden-onset left-sided flank pain. She had recently been diagnosed with systemic lupus erythematosus (SLE) and had undergone a computed tomography (CT)-guided core needle biopsy of the left kidney 9 days earlier. The results were consistent with lupus nephritis class III or IV. Initial vitals were within normal limits. She appeared pale and her left flank was tender to palpation without discoloration or abdominal distention. Laboratory investigations showed a hemoglobin level of 7.1 g/dL. The CT scan of the abdomen and pelvis revealed a large hyperdense left perinephric collection consistent with perinephric hematoma with a moderate amount of retroperitoneal stranding most prominent on the left side extending across the midline to the right side. Contrast extravasation was suspected in the lower pole of the left kidney consistent with active bleeding site. Emergent renal angiography revealed a 2 × 1 cm intraparenchymal pseudoaneurysm in the lower pole of the left kidney along with a few small microaneurysms. Coil embolization of the pseudoaneurysm was successfully performed without any complications. In conclusion, SLE or lupus nephritis in this patient may be the predisposing factors for microaneurysm and RIP formations. RIP is an unusual complication after percutaneous kidney biopsy that carries a significant mortality rate if ruptured, causing retroperitoneal hemorrhage. Clinicians should be vigilant when encountering high-risk patients with persistent hematuria, flank pain, or abdominal pain within four weeks after a kidney biopsy.


Manufacturer change and risk of system-related complications after implantable cardioverter defibrillator replacement: physicians' survey and data from the Detect Long-term Complications After Implantable Cardioverter Defibrillator Replacement Registry.
Some barriers seem to exist in changing implantable cardioverter defibrillator (ICD) manufacturer at the time of device replacement. We sought to understand the obstacles to changing ICD manufacturer within the cohort of patients enrolled in the Detect Long-term Complications After ICD Replacement Registry. We analyzed 784 consecutive ICD/cardiac resynchronization therapy defibrillator (CRT-D) device replacements within a 1.5-year time-frame in 36 Italian centers to evaluate potential factors associated with changing manufacturers and system-related complications. Manufacturer change occurred in 191 patients (24%): 72/211 single-chamber ICDs (34%), 52/210 dual-chamber ICDs (25%) and 67/363 CRT-D (18%, P < 0.0001 vs. single-chamber). Replacement-only procedures were associated with a lower rate of manufacturer change than upgrading procedures (23 vs. 32%, P = 0.02). In the single-chamber/dual-chamber cohort, the only variables associated with manufacturer change were the number of available manufacturers (OR = 1.9; P < 0.0001) and an upgrade procedure (OR = 1.7; P = 0.035), whereas the center volume was associated with maintenance of the same manufacturer (OR = 0.5; P = 0.0172). In the CRT-D group, the number of available manufacturers [OR = 2.9; P < 0.0001, service life below the median value (OR = 2.5; P = 0.0026)], and physiological design (OR = 8.4; P = 0.0048) were associated with manufacturer change. At 6-month follow-up, 17 patients (2.2%) experienced a system complication that was lead-related in all cases; upgrade procedure was the only predictor (hazard ratio = 6.7) of complications. At the time of ICD replacement, a manufacturer change occurred in 24% of patients and it was less likely in CRT-D devices, which are equipped with more specific technology and less frequently require the addition of features. System-related complications are strongly associated to upgrade procedures rather than to manufacturer change.


Widespread phytoplankton blooms triggered by 2019-2020 Australian wildfires.
Droughts and climate-change-driven warming are leading to more frequent and intense wildfires1-3, arguably contributing to the severe 2019-2020 Australian wildfires4. The environmental and ecological impacts of the fires include loss of habitats and the emission of substantial amounts of atmospheric aerosols5-7. Aerosol emissions from wildfires can lead to the atmospheric transport of macronutrients and bio-essential trace metals such as nitrogen and iron, respectively8-10. It has been suggested that the oceanic deposition of wildfire aerosols can relieve nutrient limitations and, consequently, enhance marine productivity11,12, but direct observations are lacking. Here we use satellite and autonomous biogeochemical Argo float data to evaluate the effect of 2019-2020 Australian wildfire aerosol deposition on phytoplankton productivity. We find anomalously widespread phytoplankton blooms from December 2019 to March 2020 in the Southern Ocean downwind of Australia. Aerosol samples originating from the Australian wildfires contained a high iron content and atmospheric trajectories show that these aerosols were likely to be transported to the bloom regions, suggesting that the blooms resulted from the fertilization of the iron-limited waters of the Southern Ocean. Climate models project more frequent and severe wildfires in many regions1-3. A greater appreciation of the links between wildfires, pyrogenic aerosols13, nutrient cycling and marine photosynthesis could improve our understanding of the contemporary and glacial-interglacial cycling of atmospheric CO2 and the global climate system.


Photoacoustic-Based Multimodal Nanoprobes: from Constructing to Biological Applications.
Multimodal nanoprobes have attracted intensive attentions since they can integrate various imaging modalities to obtain complementary merits of single modality. Meanwhile, recent interest in laser-induced photoacoustic imaging is rapidly growing due to its unique advantages in visualizing tissue structure and function with high spatial resolution and satisfactory imaging depth. In this review, we summarize multimodal nanoprobes involving photoacoustic imaging. In particular, we focus on the method to construct multimodal nanoprobes. We have divided the synthetic methods into two types. First, we call it "one for all" concept, which involves intrinsic properties of the element in a single particle. Second, "all in one" concept, which means integrating different functional blocks in one particle. Then, we simply introduce the applications of the multifunctional nanoprobes for in vivo imaging and imaging-guided tumor therapy. At last, we discuss the advantages and disadvantages of the present methods to construct the multimodal nanoprobes and share our viewpoints in this area.


The prevalence of self-monitoring of blood glucose and costs of glucometer strips in a nationwide cohort.
This study used nationwide data to determine the prevalence of self-monitoring of blood glucose (SMBG) among all non-institutionalized persons living in Norway and to estimate the prevalence of SMBG among diabetes patients, the frequency and cost of SMBG, and the use of different glucometers. This retrospective, descriptive study is based on data of sales of glucometer strips to non-institutionalized persons in Norway in 2008. The data included gender, age group, month of purchase, sales place, type of strips, number of packages dispensed, and cost of strips. Additionally, statistics on sales of insulin and oral antidiabetes medications were obtained from the Norwegian Prescription Database. A total of 96,999 persons purchased strips, a prevalence of 2%. Approximately 70% of diabetes patients practiced SMBG. An average patient used 1.7 strips per day, and younger patients purchased more strips than older patients. Fewer than 50% of patients performed glucose measurements daily. One percent of patients used more than 10 strips daily and was accountable for 8% of total costs. Most patients used only one type of strips, but the number of strips purchased increased with the number of different strips. The average annual cost of strips was 446 euro per person. Two percent of all non-institutionalized inhabitants and an estimated 70% of patients using diabetes medication purchased SMBG strips. A small percentage of the patients incurred a substantial proportion of the costs. This, along with the fact that over half of the patients monitor less than once per day, calls for tighter follow-up of diabetes patients.


[Immune genetic markers of anthracosilicosis].
Studies of ABO, Rh, MN, rho, Lewis blood groups in coal miners suffering from anthracosilicosis and in reference healthy individuals proved that MM phenotypes are predisposed to anthracosilicosis and MN phenotypes could be resistant to the disease. These results could be valuable for prevention of this occupational disease among coal industry workers.


Does Dexamethasone Helps in Meningococcal Sepsis?
Prompt recognition and aggressive early treatment are the only effective measures against invasive meningococcal disease (IMD). Anti-inflammatory adjunctive treatment remains controversial and difficult to assess in patients with IMD. The purpose of this study was to evaluate the effect of dexamethasone (DXM) as adjunctive treatment in different clinical forms of IMD, and attempt to answer if DXM should be routinely used in the treatment of IMD. In this non-interventional clinical study (NIS), 39 patients with meningococcal septicaemia with or without of meningitis were included, and compared regarding the impact of dexamethasone (DXM), as an adjunctive treatment, on the outcome of IMD. SPSS statistics is used for statistical processing of data. Thirty (76.9%) patients with IMD had sepsis and meningitis, and 9 (23.1%) of them had sepsis alone. Dexamethasone was used in 24 (61.5%) cases, in both clinical groups. The overall mortality rate was 10.3%. Pneumonia was diagnosed in 6 patients (15.4%), arthritis in 3 of them (7.7%), and subdural effusion in one patient (2.6%). The data showed a significant statistical difference on the length of hospitalization, and WBC normalization in groups of patients treated with DXM. The use of DXM as adjunctive therapy in invasive meningococcal disease has a degree of proven benefits and no harmful effects. In fighting this very dangerous and complex infection, even a limited benefit is sufficient to recommend the use of DXM as adjunctive treatment in invasive meningococcal disease.


Caudal epidurals: the whoosh test.
Caudal epidural injection is a simple procedure that carries a low risk of complications. The whoosh test (injection of air into the caudal epidural space with simultaneous auscultation over the thoracolumbar spine) has been recommended as an aid to correct needle placement. A 1-year prospective study, using fluoroscopic imaging to identify needle position, was conducted to compare the sensitivity and specificity of the whoosh test with that of clinical impression alone in assessing correct needle placement in the caudal space. Of 131 patients studied, correct needle placement was achieved in 121 on the first attempt (92%). Clinical impression alone had a sensitivity of 94% and a specificity of 20%. The whoosh test had a sensitivity of 80% and a specificity of 60%. The whoosh test is superior to clinical judgment in detecting incorrect caudal needle placement.


Long-term problems in patients with tracheoesophageal puncture.
Two hundred forty-seven patients, followed up a mean of 18 months and up to 108 months, were retrospectively studied for problems related to or affecting tracheoesophageal puncture for voice restoration. Active cases, as well as those inactive and decreased, were included. There were 154 patients with primary and 93 with secondary tracheoesophageal puncture. Problems were encountered at least once in 137 cases (55%). The 20 types of problems identified occurred in frequencies ranging from 0.4% to 21%. Most difficulties developed within the first 6 months and in general were easily managed. However, problems such as recalcitrant leakage, difficulty with changing the prosthesis, and dislodgment of the prosthesis led to permanent tracheoesophageal puncture tract closure in 18 patients (7%).


Comparison of US County-Level Public Health Performance Rankings With County Cluster and National Rankings: Assessment Based on Prevalence Rates of Smoking and Obesity and Motor Vehicle Crash Death Rates.
Health departments can be grouped together based on sociodemographic characteristics of the population served. Comparisons within these groups can then help with monitoring and improving the health of their populations. To compare county-level percentile rankings on outcomes of smoking, motor vehicle crash deaths, and obesity within sociodemographic peer clusters vs nationwide rankings. This cross-sectional, population-based study of demographic and health data from the 2014 Behavioral Risk Factor Surveillance System and the 2016 Robert Wood Johnson Foundation County Health Rankings data set was conducted at 3139 of 3143 US counties and county-equivalents. Four locations were excluded due to incomplete data. Data analysis was conducted between January and August 2017. Random forest algorithms were used to identify sociodemographic characteristics most associated with the outcomes of interest. These characteristics were race and ethnicity, educational attainment, age, marital status, employment status, sex, and health insurance status. k-means clustering was used to cluster counties based on these sociodemographic characteristics and the percentage of the county classified as rural. County-level smoking prevalence, motor vehicle crash death rate, and obesity prevalence. County percentile rankings on the outcomes of interest were compared in the national context and the within-cluster context. A total of 318 856 967 individuals (mean [SD] individuals per county, 101 579.2 [326 315]; 161 911 910 women [50.8%]) were represented by the 3139 counties used in this analysis. Eight distinct sociodemographic clusters throughout the United States were found. Cluster-specific percentile rankings for both smoking prevalence and motor vehicle crash death rates improved more than 70 percentile points for several counties in the rural, American Indian cluster compared with the nationwide percentiles. Conversely, the young, urban, middle to high socioeconomic status cluster included counties with cluster-specific percentile rankings that declined by 60 percentile points or more compared with the nationwide rankings for all 3 outcomes of interest. Comparing county health outcomes on a nationwide or statewide basis fails to adequately account for sociodemographic context. Clustering counties by sociodemographic factors related to the outcome of interest allows a better understanding of other factors that may be shaping the prevalence of health outcomes. These groupings may also aid learning exchange.


Association of dermal melanocytosis with lysosomal storage disease: clinical features and hypotheses regarding pathogenesis.
The potential association of dermal melanocytosis with lysosomal storage disease in infancy is an uncommonly known and poorly understood entity. We describe 2 infants with extensive dermal melanocytosis in association with GM1 gangliosidosis type 1 and Hurler syndrome, respectively. A literature analysis revealed 37 additional cases. Clinically, dermal melanocytosis associated with lysosomal storage disease is characterized by extensive, blue cutaneous pigmentation with dorsal and ventral distribution, indistinct borders, and persistent and/or "progressive" behavior. GM1 gangliosidosis type 1 and Hurler syndrome are the most common underlying disorders associated with these cutaneous features. In the appropriate clinical setting, an unusual presentation of dermal melanocytosis in an infant may be a cutaneous sign of an underlying lysosomal storage disease. The pathogenetic mechanisms behind this association remain to be elucidated.


Dissociating Value Representation and Inhibition of Inappropriate Affective Response during Reversal Learning in the Ventromedial Prefrontal Cortex.
Decision-making studies have implicated the ventromedial prefrontal cortex (vmPFC) in tracking the value of rewards and punishments. At the same time, fear-learning studies have pointed to a role of the same area in updating previously learned cue-outcome associations. To disentangle these accounts, we used a reward reversal-learning paradigm in a functional magnetic resonance imaging study in 18 human participants. Participants first learned that one of two colored squares (color A) was associated with monetary reward, whereas the other (color B) was not, and then had to learn that these contingencies reversed. Consistent with value representation, activity of a dorsal region of vmPFC was positively correlated with reward magnitude. Conversely, a more ventral region of vmPFC responded more to color A than to color B after contingency reversal, compatible with a role of inhibiting the previously learned response that was no longer appropriate. Moreover, the response strength was correlated with subjects' behavioral learning strength. Our findings provide direct evidence for the spatial dissociation of value representation and affective response inhibition in the vmPFC.


Four-year Outcomes After Cessation of Tenofovir in Immune-tolerant Chronic Hepatitis B Patients.
To study the long-term outcome after cessation of antiviral therapy in immune-tolerant patients. Experience in the treatment of immune-tolerant chronic hepatitis B is scanty. Some immune-tolerant patients may receive temporary antiviral therapy, such as for prevention of vertical transmission at pregnancy or prophylaxis for chemotherapy. This was a follow-up study of a phase 2 trial at 2 centers. Immune-tolerant patients received tenofovir disoproxil fumarate and/or emtricitabine for 4 years and were followed for another 4 years after treatment cessation. Virological relapse was defined as hepatitis B virus (HBV) DNA>2000 IU/mL; clinical relapse was defined as HBV DNA>2000 IU/mL; and alanine aminotransferase (ALT)>2 times the upper limit of normal. In total, 20 patients stopped treatment and were followed up for 206±14 weeks. All patients developed virological relapse at posttreatment week 4 (HBV DNA, 7.07±1.45 log IU/mL). A total of 10 (50%) patients developed clinical relapse at 15±11 weeks (highest ALT, 1149 U/L). In total, 11 (55%) patients were restarted on antiviral therapy; 4 achieved complete HBV DNA suppression and 1 achieved hepatitis B e antigen (HBeAg) seroconversion. Among the 9 patients not restarted on therapy, 2 patients had HBeAg seroconversion with normal ALT and HBV DNA of 7.12 and 1.62 IU/mL, respectively. The remaining 7 untreated patients continued to have positive HBeAg, high HBV DNA, and normal ALT. Rapid virological relapse is universal and clinical relapse is common after stopping antiviral therapy in patients with immune-tolerant chronic hepatitis B. HBeAg seroconversion is rare regardless of treatment reinitiation.


Urban-rural disparities in injury mortality in China, 2006.
Urban-rural disparity is an important issue for injury control in China. Details of the urban-rural disparities in fatal injuries have not been analyzed. To target key injury causes that most contribute to the urban-rural disparity, we decomposed total urban-rural differences in 2006 injury mortality by gender, age, and cause. Mortality data came from the Chinese Vital Registration data, covering a sample of about 10% of the total population. The chi-square test was used to test the significance of urban-rural disparities. For all ages combined, the injury death rate for males was 60.1/100,000 in rural areas compared with 40.9 in urban areas; for females, the respective rates were 31.5 and 23.6/100,000. The greatest disparity was at age <1 year for both sexes, where the rate from unintentional suffocation in rural areas was more than twice the urban rate. The higher mortality from drowning among males of all ages and among females ages 1-24 and 35+ contributed substantially to the age-specific urban-rural disparities. For both sexes, transportation incidents and suicide were the most important contributors to higher rates among rural residents ages 15+. Unintentional suffocation, drowning, transportation incidents, and suicide not only are the major causes of injury death, but also play a key role in explaining the urban-rural disparities in fatal injuries. Further research is needed to identify factors leading to higher rural death rates and to explore economical and feasible interventions for reducing injuries and narrowing the urban-rural gap in injury mortality.


Targeting CTLA-4: a possible solution for microsatellite-stable colorectal cancer.
Checkpoint blockade immunotherapy is a therapeutic revolution in cancer treatment. However, only 5% of patients with metastatic colorectal cancer benefit from these therapies, and these tumors genetically harbored microsatellite instability status. In contrast, tumors with stable microsatellites are considered resistant to immunotherapy, and standard treatment with chemotherapies is standard of care, with few chances of curative intent. In a recent clinical trial, we demonstrated that the combination of two chemotherapies with two immunotherapies promotes the recruitment and activation of the adaptive immune system at the tumor level, resulting in clinical benefit in a significant number of patients. In parallel, a biological study revealed biomarkers of response, including CTLA-4 expression and induction of a tumor-specific immune response.


Use of Intralipid for kinetic analysis of HDL apoC-III: evidence for a homogeneous kinetic pool of apoC-III in plasma.
Apolipoprotein C-III (apoC-III) is an important regulator of lipoprotein metabolism. Radioisotope and stable isotope kinetic studies show differing results in relation to the kinetics of apoC-III in HDL. Kinetic analysis of HDL apoC-III may be difficult because of its low concentration, as well as the presence of other apoproteins at higher concentration, in the HDL fraction. We used Intralipid(R) (IL), known to preferentially extract apoC proteins from plasma, as a means of extracting apoC-III from HDL before apoprotein separation by isoelectric focusing gel electrophoresis for the measurement of tracer enrichment. Protein purity was assessed by an isoleucine-to-leucine (Ile/Leu) ratio, as apoC-III contains no isoleucine. We compared apoC-III kinetics in 14 men using a bolus infusion of deuterated leucine. The Ile/Leu ratio for IL-extracted HDL (IL-HDL) apoC-III (3.0 +/- 0.7%) was not different from that of VLDL apoC-III (2.6 +/- 0.6%) but was significantly lower than that of untreated HDL apoC-III (9.0 +/- 2.9%) (P < 0.001). The isotopic enrichment curves and fractional catabolic rates (FCRs) for IL-HDL apoC-III were not different from those of VLDL apoC-III. In contrast, HDL apoC-III had significantly lower isotopic enrichments and FCRs than IL-HDL apoC-III (P < 0.001). In conclusion, this simple IL method can be used to isolate apoC-III from HDL with minimal interference from other HDL apoproteins, and it demonstrates that the kinetics of apoC-III in VLDL and HDL are similar, supporting the concept of a single kinetically homogeneous pool of apoC-III in plasma.


Ancient forest plants possess cytotoxic properties causing liver cancer HepG2 cell apoptosis.
Here, we collected 154 plant species in China ancient forests looking for novel efficient bioactive compounds for cancer treatments. We found 600 bioactive phyto-chemicals that induce apoptosis of liver cancer cell in vitro. First, we screen the plant extract's in vitro cytotoxicity inhibition of cancer cell growth using in vitro HepG2 cell lines and MTT cytotoxicity. The results from these initial MTT in vitro cytotoxicity tests show that the most efficient plants towards hepatoma cytoxicity is Cephalotaxus sinensis, mint bush (Elsholtzia stauntonii) and winged spindle tree (Euonymus alatus). We then used in cell-counting kit-8 (CCK-8) to further understand in vivo tumor growth using nude mice and GC-MS and LC-QTOF-MS to analyze the composition of compounds in the extracts. Extracted chemically active molecules analyzed by network pharmacology showed inhibition on the growth of liver cancer cells by acting on multiple gene targets, which is different from the currently used traditional drugs acting on only one target of liver cancer cells. Extracts from Cephalotaxus sinensis, mint bush (Elsholtzia stauntonii) and winged spindle tree (Euonymus alatus) induce apoptosis in hepatoma cancer cell line HepG2 with a killing rate of more than 83% and a tumor size decrease by 62-67% and a killing rate of only 6% of normal hepatocyte LO2. This study highlight efficient candidate species for cancer treatment providing a basis for future development of novel plant-based drugs to help meeting several of the UN SDGs and planetary health.


Directed Metabolic Pathway Evolution Enables Functional Pterin-Dependent Aromatic-Amino-Acid Hydroxylation in Escherichia coli.
Tetrahydrobiopterin-dependent hydroxylation of aromatic amino acids is the first step in the biosynthesis of many neuroactive compounds in humans. A fundamental challenge in building these pathways in Escherichia coli is the provision of the non-native hydroxylase cofactor, tetrahydrobiopterin. To solve this, we designed a genetic selection that relies on the tyrosine synthesis activity of phenylalanine hydroxylase. Using adaptive laboratory evolution, we demonstrate the use of this selection to discover: (1) a minimum set of heterologous enzymes and a host folE (T198I) mutation for achieving this type of hydroxylation chemistry in whole cells, (2) functional complementation of tetrahydrobiopterin by indigenous cofactors, and (3) a tryptophan hydroxylase mutation for improving protein abundance. Thus, the goal of having functional aromatic-amino-acid hydroxylation in E. coli was achieved through directed metabolic pathway evolution.


Assessing the Ability of Salmonella enterica to Translocate Type III Effectors Into Plant Cells.
Salmonella enterica serovar Typhimurium, a human enteric pathogen, has the ability to multiply and survive endophytically in plants. Genes encoding the type III secretion system (T3SS) or its effectors (T3Es) may contribute to its colonization. Two reporter plasmids for T3E translocation into plant cells that are based on hypersensitive response domains of avirulence proteins from the Pantoea agglomerans-beet and Xanthomonas euvesicatoria-pepper pathosystems were employed in this study to investigate the role of T3Es in the interaction of Salmonella ser. Typhimurium 14028 with plants. The T3Es of Salmonella ser. Typhimurium, SipB and SifA, which are translocated into animal cells, could not be delivered by Salmonella ser. Typhimurium into cells of beet roots or pepper leaves. In contrast, these effectors were translocated into plant cells by the phytopathogenic bacteria P. agglomerans pv. betae, Erwinia amylovora, and X. euvesicatoria. Similarly, HsvG, a T3E of P. agglomerans pv. gypsophilae, and XopAU of X. euvesicatoria could be translocated into beet roots and pepper leaves, respectively, by the plant pathogens but not by Salmonella ser. Typhimurium. Mutations in Salmonella ser. Typhimurium T3SS genes invA, ssaV, sipB, or sifA, did not affect its endophytic colonization of lettuce leaves, supporting the notion that S. enterica cannot translocate T3Es into plant cells.


Multi-gas sensing based on photoacoustic spectroscopy using tunable laser diodes.
Multi-hydrogenated compounds detection based on photoacoustic (PA) spectroscopy is reported. Three near-infrared semiconductor lasers are used with a resonant PA cell operated in its first longitudinal mode to monitor methane, water vapour and hydrogen chloride in the parts per million range. The design of our cell results from simulations performed in order to optimise its performances. Influence of the buffer gas on the PA signal has also been analysed, both theoretically and experimentally. A reduction of the PA signal of almost one order of magnitude has been observed between N(2) and He, which demonstrates the importance of the buffer gas in PA spectroscopy. Finally, detection limits of 0.5 ppm of CH(4) and 3 ppm of HCl has been achieved experimentally in nitrogen and an H(2)O sensitivity of 0.2 ppm has been estimated.


Concentration of total microcystins associates with nitrate and nitrite, and may disrupt the nitrogen cycle, in warm-monomictic lakes of the southcentral United States.
Cyanobacterial blooms and the toxins they produce pose a growing threat worldwide. Mitigation of such events has primarily focused on phosphorus management and has largely neglected the role of nitrogen. Previous bloom research and proposed management strategies have primarily focused on temperate, dimictic lakes, and less on warm-monomictic systems like those at subtropical latitudes. The in-lake conditions, concentration of total microcystins, and microbial functioning of twenty warm-monomictic lakes in the southcentral United States were explored in the spring and summer of 2021. Our data revealed widespread microcystins in lakes across this region, some of which exceeded regulatory limits. Microcystins were higher in the spring compared to the summer, indicating that warm-monomictic lakes, even across a large range of precipitation, do not follow the trends of temperate dimictic lakes. Microcystins were found in surface waters and bottom waters well below the photic zone, reflecting the persistence of these toxins in the environment. Principal components analyses showed a strong association between microcystins, nitrate + nitrite, and Planktothrix relative abundance and transcriptional activity. Many systems exhibited stronger denitrification in the spring, perhaps contributing to the decreased toxin concentrations in the summer. Counter to most sampled lakes, one lake with the highest concentration of total microcystins indicated nitrogen cycle disruption, including inhibited denitrification. These findings are relevant to mitigating cyanobacterial blooms and toxin production in warm-monomictic systems, and suggests a need to consider nitrogen, and not solely phosphorus, in nutrient management discussions.


The proteasome inhibitor bortezomib sensitizes cells to killing by death receptor ligand TRAIL via BH3-only proteins Bik and Bim.
Previously, we showed that the proteasome inhibitor bortezomib/Velcade (formerly PS-341) synergizes with the protein tumor necrosis factor alpha-related apoptosis-inducing ligand (TRAIL), a ligand for certain death receptors, to induce apoptosis in cell lines derived from prostate and colon cancers. Because apoptosis is often triggered by BH3-only proteins of the Bcl-2 family, we have explored the hypothesis that bortezomib contributes to the apoptosis by up-regulating their levels. Indeed, bortezomib induced increases of Bik and/or Bim in multiple cell lines but not notably of two other BH3-only proteins (Puma and Bid) nor other family members (Bax, Bak, Bcl-2, and Bcl-xL). The increase in Bik levels seems to reflect inhibition by bortezomib of its proteasome-mediated degradation. Importantly, both Bik and Bim seem central to the proapoptotic function of bortezomib, because mouse embryo fibroblasts in which the genes for both Bik and Bim had been disrupted were refractory to its cytotoxic action. Similarly, the synergy between bortezomib and TRAIL in killing human prostate cancer cells was impaired in cells in which both Bik and Bim were down-regulated by RNA interference. Further evidence that bortezomib acts through the mitochondrial pathway regulated by the Bcl-2 family is that deficiency for APAF-1, which acts downstream of Bcl-2, also blocked its apoptotic effect. These results implicate BH3-only proteins, in particular both Bik and Bim, as important mediators of the antitumor action of bortezomib and establish their role in its enhancement of TRAIL-induced apoptosis.


Optimizing postoperative pain management.
The quality of postoperative pain management can be improved. Although many safe and effective therapies exist, their utilization varies considerably between and within institutions. Major challenges include the appropriate prescribing of analgesic therapies and the timely response to suboptimal pain control. Patients' satisfaction with their analgesic care may depend less on how well their pain is controlled and more on the attentiveness of their caregivers.


Hemosuccus pancreaticus complicating chronic pancreatitis: an obscure cause of upper gastrointestinal bleeding.
Hemosuccus pancreaticus, a rare form of upper gastrointestinal bleeding, may complicate chronic pancreatitis and pose a significant diagnostic and therapeutic dilemma. To present our experience with this potentially life-threatening complication of chronic pancreatitis. We reviewed our experience with management (both operative as well as angiographic embolization) of patients with hemosuccus pancreaticus complicating histologically documented chronic pancreatitis between 1976 and 1997. Diagnosis of hemosuccus pancreaticus was based on clinical presentation, preoperative endoscopic and radiographic imaging, operative findings, and pathologic evaluation. During the period, we managed eight patients with hemosuccus pancreaticus (1.5% of all patients with chronic pancreatitis treated surgically). Gastrointestinal bleeding presented as hematemesis in three and hematochezia in three, but all had recent melena and were anemic; three of these patients were hemodynamically unstable. Abdominal pain was present in six. When performed, angiography (n=6) was diagnostic of a pseudoaneurysm; computed tomography (n=7) showed a pseudoaneurysm in two and a pseudocyst in five. Endoscopy (n=8) revealed blood issuing from the ampullary papilla in two patients. Operative management (n=6) involved distal pancreatectomy, pancreatoduodenectomy, or total pancreatectomy in two patients each. Angiographic embolization was successful in one patient, but the other died from uncontrollable hemorrhage. Hemosuccus pancreaticus is rare, but should be considered in patients with chronic pancreatitis and gastrointestinal bleeding. In the absence of pancreatitis-related indications for surgery, angiographic embolization can be definitive treatment. If there are pancreatitis-related indications for operation, angiographic embolization may allow an elective operative procedure based on structural changes of the pancreas. If embolization fails, pancreatic resection is usually required, often on an emergent basis.


Acute stress in residents during emergency care: a study of personal and situational factors.
Providing care for simulated emergency patients may induce considerable acute stress in physicians. However, the acute stress provoked in a real-life emergency room (ER) is not well known. Our aim was to assess acute stress responses in residents during real emergency care and investigate the related personal and situational factors. A cross-sectional observational study was carried out at an emergency department of a tertiary teaching hospital. All second-year internal medicine residents were invited to voluntarily participate in this study. Acute stress markers were assessed at baseline (T1), before residents started their ER shift, and immediately after an emergency situation (T2), using heart rate, systolic, and diastolic blood pressure, salivary α-amylase activity, salivary interleukin-1 β, and the State-Trait Anxiety Inventory (STAI-s and STAI-t). Twenty-four residents were assessed during 40 emergency situations. All stress markers presented a statistically significant increase between T1 and T2. IL-1 β presented the highest percent increase (141.0%, p < .001), followed by AA (99.0%, p = .002), HR (81.0%, p < .001), DBP (8.0%, p < .001), and SBP (3.0%, p < .001). In the multivariable analysis, time of residency had a negative correlation with HR during the emergency (adjusted R-square = .168; F = 8.69; p = .006), SBP response (adjusted R-square = .210; F = 6.19; p = .005) and DBP response (adjusted R-square = .293; F = 9.09; p = .001). Trait anxiety (STAI-t) was positively correlated with STAI-s (adjusted R-square = .326; F = 19.9; p < .001), and number of procedures performed during emergency care had a positive association with HR response (adjusted R-square = .241; F = 5.02; p = .005). In the present study, emergency care provoked substantial acute stress in residents. Resident experience, trait anxiety, and number of emergency procedures were independently associated with acute stress response.


Resetting of exaggerated tubuloglomerular feedback activity in acutely volume-expanded young SHR.
One purpose of the present study was to evaluate the ability of 7-wk-old spontaneously hypertensive rats (SHR) to reset tubuloglomerular feedback (TGF) activity in response to acute volume expansion (VE). Second, we evaluated the contribution of ANG II, via its action on AT1 receptors, to TGF control of glomerular function during VE. TGF was assessed by micropuncture methods and proximal tubular stop-flow pressure (SFP) determinations in SHR, Wistar-Kyoto rats (WKY), and Sprague-Dawley rats (SD). During euvolemia SHR exhibited enhanced TGF activity. In the same animals acute VE was achieved by infusion of saline (5 ml. h-1. 100 g body wt-1). VE led to resetting of TGF in all three strains. Maximal SFP responses, elicited by a 30-40 nl/min loop of Henle perfusion rate, decreased from 19 to 12 mmHg in SHR and, on average, from 11 to 5 mmHg in WKY and SD (P < 0.001). Tubular flow rate producing a half-maximal response (turning point) shifted to higher flow rates during VE, from 12 to 14 nl/min in SHR and from 15 to 19 nl/min in WKY. Administration of the AT1 receptor blocker candesartan (0.05 mg/kg iv) during sustained VE decreased TGF-mediated reductions in SFP in SHR and slightly increased the turning point in WKY. Nevertheless, other parameters of TGF activity were unaffected by AT1 receptor blockade. In conclusion, young SHR possess the ability to reset TGF activity in response to VE to a degree similar to compensatory adjustments in WKY. However, TGF remains enhanced in SHR during VE. ANG II and its action on AT1 receptors are in part responsible for the exaggerated SFP responses in young SHR during VE.


Maternal diabetes modulates renal morphogenesis in offspring.
Maternal diabetes leads to an adverse in utero environment, but whether maternal diabetes impairs nephrogenesis is unknown. Diabetes was induced with streptozotocin in pregnant Hoxb7-green fluorescence protein mice at embryonic day 13, and the offspring were examined at several time points after birth. Compared with offspring of nondiabetic controls, offspring of diabetic mice had lower body weight, body size, kidney weight, and nephron number. The observed renal dysmorphogenesis may be the result of increased apoptosis, because immunohistochemical analysis revealed significantly more apoptotic podocytes as well as increased active caspase-3 immunostaining in the renal tubules compared with control mice. Regarding potential mediators of these differences, offspring of diabetic mice had increased expression of intrarenal angiotensinogen and renin mRNA, upregulation of NF-kappaB isoforms p50 and p65, and activation of the NF-kappaB pathway. In conclusion, maternal diabetes impairs nephrogenesis, possibly via enhanced intrarenal activation of the renin-angiotensin system and NF-kappaB signaling.


Improved Multi-Head Self-Attention Classification Network for Multi-View Fetal Echocardiography Recognition.
The accurate acquisition of multiview fetal cardiac ultrasound images is very important for the diagnosis of fetal congenital heart disease (FCHD). However, these manual clinical procedures have drawbacks, e.g., varying technical capabilities and inefficiency. Therefore, exploring automatic recognition method for multiview images of fetal heart ultrasound scans is highly desirable to improve prenatal diagnosis efficiency and accuracy. In this work, we propose an improved multi-head self-attention mechanism called IMSA combined with residual networks to stably solve the problem of multiview identification and anatomical structure localization. In details, IMSA can capture short- and long-range dependencies from different subspaces and merge them to extract more precise features, thus making use of the correlation between fetal heart structures to make view recognition more focused on anatomical structures rather than disturbing regions, such as artifacts and speckle noises. We validate our proposed method on fetal cardiac ultrasound imaging datasets from a single center and 38 multicenter studies and the results outperform other state-of-the-art networks by 3%-15% of F1 scores in fetal heart six standard view recognition.Clinical Relevance- This technology has great potential in assisting cardiologists to complete the automatic acquisition of multi-section fetal echocardiography images.


Ultrasound Imaging of Muscle Contraction of the Tibialis Anterior in Patients with Facioscapulohumeral Dystrophy.
A need exists for biomarkers to diagnose, quantify and longitudinally follow facioscapulohumeral muscular dystrophy (FSHD) and many other neuromuscular disorders. Furthermore, the pathophysiological mechanisms leading to muscle weakness in most neuromuscular disorders are not completely understood. Dynamic ultrasound imaging (B-mode image sequences) in combination with speckle tracking is an easy, applicable and patient-friendly imaging tool to visualize and quantify muscle deformation. This dynamic information provides insight in the pathophysiological mechanisms and may help to distinguish the various stages of diseased muscle in FSHD. In this proof-of-principle study, we applied a speckle tracking technique to 2-D ultrasound image sequences to quantify the deformation of the tibialis anterior muscle in patients with FSHD and in healthy controls. The resulting deformation patterns were compared with muscle ultrasound echo intensity analysis (a measure of fat infiltration and dystrophy) and clinical outcome measures. Of the four FSHD patients, two patients had severe peroneal weakness and two patients had mild peroneal weakness on clinical examination. We found a markedly varied muscle deformation pattern between these groups: patients with severe peroneal weakness showed a different motion pattern of the tibialis anterior, with overall less displacement of the central tendon region, while healthy patients showed a non-uniform displacement pattern, with the central aponeurosis showing the largest displacement. Hence, dynamic muscle ultrasound of the tibialis anterior muscle in patients with FSHD revealed a distinctively different tissue deformation pattern among persons with and without tibialis anterior weakness. These findings could clarify the understanding of the pathophysiology of muscle weakness in FSHD patients. In addition, the change in muscle deformation shows good correlation with clinical measures and quantitative muscle ultrasound measurements. In conclusion, dynamic ultrasound in combination with speckle tracking allows the study of the effects of muscle pathology in relation to strength, force transmission and movement generation. Although further research is required, this technique can develop into a biomarker to quantify muscle disease severity.


Wrist accelerometry for monitoring dementia agitation behaviour in clinical settings: A scoping review.
Agitated behaviour among elderly people with dementia is a challenge in clinical management. Wrist accelerometry could be a versatile tool for making objective, quantitative, and long-term assessments. The objective of this review was to summarise the clinical application of wrist accelerometry to agitation assessments and ways of analysing the data. Two authors independently searched the electronic databases CINAHL, PubMed, PsycInfo, EMBASE, and Web of Science. Nine (n = 9) articles were eligible for a review. Our review found a significant association between the activity levels (frequency and entropy) measured by accelerometers and the benchmark instrument of agitated behaviour. However, the performance of wrist accelerometry in identifying the occurrence of agitation episodes was unsatisfactory. Elderly people with dementia have also been monitored in existing studies by investigating the at-risk time for their agitation episodes (daytime and evening). Consideration may be given in future studies on wrist accelerometry to unifying the parameters of interest and the cut-off and measurement periods, and to using a sampling window to standardise the protocol for assessing agitated behaviour through wrist accelerometry.


The small heat-shock protein αB-crystallin is essential for the nuclear localization of Smad4: impact on pulmonary fibrosis.
Idiopathic pulmonary fibrosis (IPF) is a devastating disease characterized by the proliferation of myofibroblasts and the accumulation of extracellular matrix (ECM) in the lungs. TGF-β1 is the major profibrotic cytokine involved in IPF and is responsible for myofibroblast proliferation and differentiation and ECM synthesis. αB-crystallin is constitutively expressed in the lungs and is inducible by stress, acts as a chaperone and is known to play a role in cell cytoskeleton architecture homeostasis. The role of αB-crystallin in fibrogenesis remains unknown. The principal signalling pathway involved in this process is the Smad-dependent pathway. We demonstrate here that αB-crystallin is strongly expressed in fibrotic lung tissue from IPF patients and in vivo rodent models of pulmonary fibrosis. We also show that αB-crystallin-deficient mice are protected from bleomycin-induced fibrosis. Similar protection from fibrosis was observed in αB-crystallin KO mice after transient adenoviral-mediated over-expression of IL-1β or TGF-β1. We show in vitro in primary epithelial cells and fibroblasts that αB-crystallin increases the nuclear localization of Smad4, thereby enhancing the TGF-β1-Smad pathway and the consequent activation of TGF-β1 downstream genes. αB-crystallin over-expression disrupts Smad4 mono-ubiquitination by interacting with its E3-ubiquitin ligase, TIF1γ, thus limiting its nuclear export. Conversely, in the absence of αB-crystallin, TIF1γ can freely interact with Smad4. Consequently, Smad4 mono-ubiquitination and nuclear export are favoured and thus TGF-β1-Smad4 pro-fibrotic activity is inhibited. This study demonstrates that αB-crystallin may be a key target for the development of specific drugs in the treatment of IPF or other fibrotic diseases.


Introduction of an antifouling photoelectrode: an effective strategy for a high-performance photoelectrochemical cytosensor.
As nonspecific adsorption or biofouling has obvious side effects on the selectivity, it is a great challenge for cytosensors to detect target cells in practical biological samples. In this study, we first propose the design and synthesis of an antifouling photoelectrode. The antifouling photoelectrode not only has the desired photocurrent response, but also possesses an unexpected antifouling capability of resisting nonspecific adsorption of biomolecules. Herein, the PEDOT-HPG/SnS/ZnO-NT antifouling photoelectrode was formed and a robust photoelectrochemical cytosensor with enhanced sensitivity and selectivity has been demonstrated.


Rivastigmine transdermal patch and capsule in Alzheimer's disease: influence of disease stage on response to therapy.
The cholinesterase inhibitor rivastigmine is approved for the symptomatic treatment of mild to moderate Alzheimer's disease (AD). This exploratory, hypothesis-forming analysis assessed response to rivastigmine according to severity of dementia at baseline. This was a retrospective analysis of a large randomized, placebo-controlled trial (ENA713D2320). AD patients treated with 9.5 mg/24 h rivastigmine patch, 17.4 mg/24 h rivastigmine patch, rivastigmine capsule (12 mg/day), or placebo were stratified according to baseline Mini-Mental State Examination (MMSE) scores: ≥7 to ≤12 (severe disease), ≥13 to ≤15 (moderately severe), ≥16 to ≤18 (moderate), or ≥19 to ≤25 (mild to moderate). Changes from baseline at Week 24 on Alzheimer's Disease Assessment Scale-cognitive subscale (ADAS-cog), Alzheimer's Disease Cooperative Study-Clinical Global Impression of Change (ADCS-CGIC), and Alzheimer's Disease Cooperative Study-Activities of Daily Living (ADCS-ADL) were assessed. Based on baseline MMSE scores, 141, 228, 333, and 348 patients had severe, moderately severe, moderate, and mild to moderate dementia. Worsening of ADAS-cog, ADCS-CGIC, and ADCS-ADL scores in patients receiving placebo were greater in patients with more severe dementia. Significant improvements versus placebo were seen with rivastigmine patch and/or capsule on ADAS-cog, ADCS-CGIC, and ADCS-ADL scores in patients with severe, moderately severe, and moderate AD (all p < 0.05). However, no significant improvements were seen in rivastigmine-treated patients with mild to moderate AD. Rivastigmine benefits AD patients across dementia stages. Similar to previous cholinesterase inhibitor studies, greatest treatment effects with rivastigmine patch and capsule were seen in patients with more advanced dementia, most likely driven by greater placebo decline in this population.


TYMSTR, a putative chemokine receptor selectively expressed in activated T cells, exhibits HIV-1 coreceptor function.
Chemokines bind to specific receptors and mediate leukocyte migration to sites of inflammation. Recently, some chemokine receptors, notably CXCR4 and CCR5, have been shown to be essential fusion factors on target cells for infection by human immunodeficiency virus (HIV); the chemokines bound by these receptors have also been shown to act as potent inhibitors of HIV infection. Here, we describe the isolation of a novel, putative chemokine receptor. We have isolated the cDNA for a putative human chemokine receptor, which we have termed TYMSTR (T-lymphocyte-expressed seven-transmembrane domain receptor). The TYMSTR gene is localized to human chromosome 3 and encodes a protein that has a high level of identity with chemokine receptors. TYMSTR mRNA was selectively expressed in interleukin-2-stimulated T lymphocytes but not in freshly isolated lymphocytes and leukocytes or related cell lines. The natural ligand for TYMSTR was not identified among 32 human chemokines and other potential ligands. Cells co-expressing TYMSTR and human CD4 fused with cells expressing envelope glycoproteins of macrophage (M)-tropic HIV-1 as well as T-cell line (T)-tropic HIV-1 isolates. Addition of infectious, T-tropic HIV-1 particles to TYMSTR/CD4-expressing cells resulted in viral entry and proviral DNA formation. Our findings demonstrate that TYMSTR, in combination with CD4, mediates HIV-1 fusion and entry. The high-level expression of TYMSTR in CD4(+) T lymphocytes and the selectivity of this receptor for T-tropic and M-tropic HIV-1 strains indicates that TYMSTR might function as HIV coreceptor at both early and late stages of infection.


A study in male and female 5-HT transporter knockout rats: an animal model for anxiety and depression disorders.
Human studies have shown that a reduction of 5-HT transporter (SERT) increases the vulnerability for anxiety and depression. Moreover, women are more vulnerable to develop depression and anxiety disorders than men. For that reason we hypothesized that homozygous 5-HT transporter knockout rat (SERT(-/-)) models, especially female, are valuable and reliable animal models for humans with an increased vulnerability for anxiety- and depression-related disorders. As rats are extensively used in neuroscience research, we used the unique 5-HT transporter knockout rat, that was recently generated using N-ethyl-N-nitrosurea (ENU) -driven mutagenesis, to test this hypothesis. Behavioral testing revealed that male and female SERT(-/-) rats spent less time in the center of the open field and spent less time on the open arm of the elevated plus maze compared with wild-type 5-HT transporter knockout rats (SERT(+/+)). In the novelty suppressed feeding test, only male SERT(-/-) rats showed a higher latency before starting to eat in a bright novel arena compared with SERT(+/+) controls. Both male and female SERT(-/-) rats showed a higher escape latency from their home cage than SERT(+/+) littermates. Moreover, SERT(-/-) rats were less mobile in the forced swim test, and sucrose consumption was reduced in SERT(-/-) rats relative to SERT(+/+) rats. Both effects were sex-independent. Neurochemically, basal extracellular 5-HT levels were elevated to a similar extent in male and female SERT(-/-) rats, which was not influenced by the selective 5-HT reuptake inhibitor citalopram. 5-HT immunostaining revealed no difference between SERT(+/+) and SERT(-/-) rats in the dorsal raphe nuclei, in both males and females. These findings demonstrate that SERT(-/-) rats show anxiety and depression-related behavior, independent of sex. Genetic inactivation of the SERT has apparently such a great impact on behavior, that hardly any differences are found between male and female rats. This knockout rat model may provide a valuable model to study anxiety- and depression-related disorders in male and female rats.


Effect of Introduced Pseudomonas fluorescens Strains on Soil Nematode and Protozoan Populations in the Rhizosphere of Wheat and Pea.
Previous studies have shown that inoculation of pea seeds with Pseudomonas fluorescens strains F113lacZY or F113G22 increased mineralization of organic nitrogen in the rhizosphere. In contrast, inoculation of the same strains onto wheat seeds reduced mineralization of N from organic residues incorporated into soil. In the present study, we report on a likely explanation of this phenomenon, which appears to be governed by the effect of plant-microbe interactions on bacterial-feeding nematodes and protozoa. In soil microcosm tests, inoculation of pea seeds with Pseudomonas fluorescens strains F113lacZY or F113G22 resulted in an increase in the number of nematodes and protozoa in the rhizosphere as compared to noninoculated controls. This trend was repeated using a model sand system into which the bacteriophagous nematode Caenorhabditis elegans was introduced. It was subsequently found that non-inoculated germinating pea seeds exerted a nematicidal effect on C. elegans, which was remedied by inoculation with either strain F113lacZY or F113G22. This suggests that nematicidal compounds released by the germinating pea seeds were metabolized by the microbial inoculants before they affected nematode populations in the spermosphere or rhizosphere of pea. In contrast, inoculation of wheat plants resulted in significantly lower nematode populations in the rhizosphere, whereas protozoan numbers were unaffected. No nematicidal effects of inoculated or noninoculated wheat seeds could be found, suggesting that microfaunal populations were affected at a later stage during plant growth. Because of their key roles in accelerating the turnover of microbially immobilized N and organic matter, plants that support a larger microfaunal population are likely to benefit from a higher availability of inorganic nitrogen. Therefore, an understanding of plant-microbe interactions and their effects on soil microfaunal populations is essential in order to assess the effects of microbial inocula on plant mineral nutrition.


The incidence and importance of bacterial contaminants of cadaveric renal perfusion fluid.
Infections represent a significant risk in the postoperative transplant recipient. The perfusion fluid used to perfuse and preserve the kidneys prior to transplantation represents a potential medium in which organisms can grow. The aim of this study was to determine the incidence and clinical relevance of bacterial contamination of perfusion fluid. A total of 4 centres participated in the study and 269 perfusion fluid samples were taken for microbiological analysis. Organisms were isolated from 38 out of 218 (17.4%) perfusion fluid samples taken prior to allograft implantation and 23 out of 51 (45%) samples taken at procurement. Low virulence organisms predominated although Staphylococcus aureus, Pseudomonas aeruginosa and Escherichia coli were also isolated. Although infective complications were not seen in the allograft recipients, given the frequency with which contamination occurs and the variation in unit antibiotic protocols, we recommend the routine culturing of perfusion fluid to ensure that any potentially significant organisms are identified and treated appropriately.


Treatment of multiple myeloma and macroglobulinemai Waldenström: A long-term follow-up study.
Fifteen cases of multiple myeloma and 6 cases of macroglobulin emia Waldenstrom were followed up from 1957 to 1974. As for administration of drugs a low continuous dose regimen was mainly employed instead of a high intermittent dose regimen. 50% survival time from the onset of the disease was 18 months for multiple myeloma and 25 months for macroglobulinemia Waldenstrom. 3 cases of multiple myeloma are still living 44 months after the onset of symptoms. Cyclophosphamide and melphalan seem to have contributed much to the prolonged survival of these patients as well as improved supportive care.


Why don't neurosurgery patients return for neuropsychological follow-up? Predictors for voluntary appointment keeping and reasons for cancellation.
Neuropsychological follow-up appointments are important for patients who have had intracranial surgery because cognitive deficits are common in this population and prognosis is not always optimistic. Unfortunately some patients cancel or do not show up. The current study attempted to identify predictors of non-attendance in this population. A total of 428 patients recruited over 2 years with a scheduled neuropsychological follow-up appointment after intracranial surgery in the St. Elisabeth Hospital, Tilburg, The Netherlands were included. Demographic, clinical, and other miscellaneous variables were extracted from medical records. Of this total population, 42% were non-attenders. The predictors of non-attendance were as follows: patients who had subdural hematomas and/or malignant tumors (compared to those who had other diagnoses prior to intracranial surgery); those who had been transferred to another hospital (compared to those sent home); those who had been referred for further medical treatment before the appointment; a shorter time interval between discharge and follow-up appointment; and finally, if the patient's home was further away from the hospital. Patients who undergo intracranial surgery are a very heterogeneous group with different needs. Neuropsychological follow-up after surgery may be important for some patients (the better-functioning and/or those with cognitive complaints) but perhaps not for others (those with more severe prognoses and/or no complaints). We provide suggestions which should increase attendance in those who could benefit from follow-up neuropsychological assessment.


Virtual reality for animal navigation with camera-based optical flow tracking.
Virtual reality combined with a spherical treadmill is used across species for studying neural circuits underlying navigation and learning. We developed an optical flow-based method for tracking treadmill ball motion in real time using a single high-resolution camera. Tracking accuracy and timing were determined using calibration data. Ball tracking was performed at 500 Hz and integrated with an open source game engine for virtual reality projection. The projection was updated at 120 Hz with a latency with respect to ball motion of 30 ± 8 ms. The system was tested for behavior with fruit flies. The application and source code are available at https://github.com/ivan-vishniakou/neural-circuits-vr. Optical flow-based tracking of treadmill motion is typically achieved using optical mice. The camera-based optical flow tracking system developed here is based on off-the-shelf components and offers control over the image acquisition and processing parameters. This results in flexibility with respect to tracking conditions - such as ball surface texture, lighting conditions, or ball size - as well as camera alignment and calibration. A fast system for rotational ball motion tracking suitable for virtual reality behavior with fruit flies was developed and characterized.