Functional Activity of Matrix Metalloproteinases 2 and 9 in Tears of Patients With Glaucoma.
To evaluate the differential expression of tear matrix metalloproteinases (MMP) 2 and 9 in of patients with various forms of glaucoma. Tear samples were collected with a Schirmer's strip from 148 eyes of 113 patients (medically naïve patients with primary open-angle [POAG] or angle closure glaucoma [PACG] and those with pseudoexfoliation syndrome [PXF] or glaucoma [PXG]). These were compared to patients undergoing cataract surgery (controls) for this cross-sectional study. Functional activities of tear MMP-9 and MMP-2 were analyzed by gelatin zymography. Tenon's capsules (n = 15) were harvested from the inferior quadrant in those undergoing cataract surgery and protein expression of MMP-9 was analyzed by immunohistochemistry (IHC). Hydrogen peroxide (H2O2) stress-induced effects on in vitro activities of MMP-9 in human trabecular meshwork (HTM) cells were analyzed. The MMP-9 activity in tears was increased significantly in POAG, (n = 27), PACG (n = 24), and PXF (n = 40) eyes compared to controls (n = 35), and was increased significantly in eyes with glaucoma compared to moderate/severe glaucoma (P < 0.001). The MMP-9 expression was significantly lower in PXG (n = 22) eyes. Immunohistochemistry of Tenon's capsule revealed increased expression of MMP-9 in primary glaucoma eyes. Increased MMP-9 activity was seen in in vitro by gelatin zymography and was confirmed by Western and immunofluorescent assay on HTM upon 800 and 1000 μM H2O2-induced stress for 2 to 3 hours with approximately 80% cell death. Increased tear MMP-9 activity in early glaucoma and pseudoexfoliation syndrome suggesting activation of extracellular matrix (ECM) degradation can be used as a tear-based predictive biomarker. Decreased expression in advanced stages suggests exhaustion of the degradation response.


SERS Sensors: Recent Developments and a Generalized Classification Scheme Based on the Signal Origin.
Owing to its extreme sensitivity and easy execution, surface-enhanced Raman spectroscopy (SERS) now finds application for a wide variety of problems requiring sensitive and targeted analyte detection. This widespread application has prompted a proliferation of different SERS-based sensors, suggesting the need for a framework to classify existing methods and guide the development of new techniques. After a brief discussion of the general SERS modalities, we classify SERS-based sensors according the origin of the signal. Three major categories emerge from this analysis: surface-affinity strategy, SERS-tag strategy, and probe-mediated strategy. For each case, we describe the mechanism of action, give selected examples, and point out general misconceptions to aid the construction of new devices. We hope this review serves as a useful tutorial guide and helps readers to better classify and design practical and effective SERS-based sensors.


Rapid induction of clinical response with a short-term high-dose starting schedule of budesonide nebulizing suspension in young children with recurrent wheezing episodes.
There are no data currently available on the correct schedule for the initiation of treatment with nebulized suspension of budesonide in children with recurrent wheezing episodes. We compared the efficacy and safety of starting with a high dose followed by a stepwise decrease to a continuous low dose. In a double-blind design, 42 children aged 6 months to 3 years were randomly allocated to receive either a high starting dose of 1 mg budesonide twice daily followed by a stepwise decrease of 25% every second day for 1 week (group A) or a low dose of 0.25 mg twice daily for 1 week (group B). Efficacy was assessed with daily symptom scores and the systemic effect of the corticosteroids with the adrenocorticotropic hormone test. The two groups were comparable for all parameters evaluated. During the first week of treatment, there was a significant decrease in asthmatic symptomatology only in group A: a 59% decrease for wheezing (p = 0.0001), 39% for diurnal cough (p = 0.036), and 39% for nocturnal cough (p = 0.04). Mean time to clinical response was 3.0 days in group A and 5.7 days in group B (p = 0.02). This early improvement was sustained for the rest of the follow-up period. The high dose starting schedule was not associated with any change in serum cortisol level. The administration of nebulized suspension of budesonide at a high starting dose schedule followed by a rapid (1 week) stepwise decrease yields a significant early improvement in asthma symptoms and causes no change in serum cortisol levels.


Heat capacity and entropy changes of calmodulin induced by calcium binding.
Microcalorimetric titrations were carried out in order to analyze the structural changes of calmodulin caused by Ca2+-binding. Measurements were made both in the absence and in the presence of Mg2+, at 5, 15, and 25 degrees C. Titrations of calmodulin with Ca2+ in the absence of Mg2+ showed that the Ca2+-binding reaction is endothermic and thus is driven solely by the large entropy change. Following the method of Sturtevant (1977), the magnitudes of the hydrophobic and intramolecular vibrational contributions to the heat capacity and entropy changes of calmodulin on Ca2+-binding were estimated. In both the absence and the presence of Mg2+, Ca2+-binding to every site of calmodulin gives rise to an increase in hydrophobic entropy and thus to an "assembling" of nonpolar groups, which are scattered on the surface of the molecule in Ca2+-free calmodulin. Ca2+-binding to calmodulin also gives rise to an increase in vibrational entropy, indicating a "softening" of the overall structure.


Combinations and Temporal Associations Among Precursor Symptoms Before a First Episode of Psychosis.
Symptoms that precede a first episode of psychosis (FEP) can ideally be targeted by early intervention services with the aim of preventing or delaying psychosis onset. However, these precursor symptoms emerge in combinations and sequences that do not rest fully within traditional diagnostic categories. To advance our understanding of illness trajectories preceding FEP, we aimed to investigate combinations and temporal associations among precursor symptoms. Participants were from PEPP-Montréal, a catchment-based early intervention program for FEP. Through semistructured interviews, collateral from relatives, and a review of health and social records, we retrospectively measured the presence or absence of 29 precursor symptoms, including 9 subthreshold psychotic and 20 nonpsychotic symptoms. Sequences of symptoms were derived from the timing of the first precursor symptom relative to the onset of FEP. The sample included 390 participants (68% men; age range: 14-35 years). Combinations of precursor symptoms most frequently featured depression, anxiety, and substance use. Of 256 possible pairs of initial and subsequent precursor symptoms, many had asymmetrical associations: eg, when the first symptom was suspiciousness, the incidence rate ratio (IRR) of subsequent anxiety was 3.40 (95% confidence interval [CI]: 1.79, 6.46), but when the first symptom was anxiety, the IRR of subsequent suspiciousness was 1.15 (95% CI: 0.77, 1.73). A detailed examination of precursor symptoms reveals diverse clinical profiles that cut across diagnostic categories and evolve longitudinally prior to FEP. Their identification may contribute to risk assessments and provide insights into the mechanisms of illness progression.


EthSEQ: ethnicity annotation from whole exome sequencing data.
Whole exome sequencing (WES) is widely utilized both in translational cancer genomics studies and in the setting of precision medicine. Stratification of individual's ethnicity is fundamental for the correct interpretation of personal genomic variation impact. We implemented EthSEQ to provide reliable and rapid ethnicity annotation from whole exome sequencing individual's data, validated it on 1000 Genome Project and TCGA data (2700 samples) demonstrating high precision, and finally assessed computational performances compared to other tools. EthSEQ can be integrated into any WES based processing pipeline and exploits multi-core capabilities. R package available at github.com/aromanel/EthSEQ and CRAN repository. firstname.lastname@example.com or email@example.com. Supplementary data are available at Bioinformatics online.


Mechanisms Underlying Chronic Binge Alcohol Exposure-Induced Uterine Artery Dysfunction in Pregnant Rat.
A cardinal feature of fetal alcohol syndrome is growth restriction. Maternal uterine artery adaptations to pregnancy correlate with birthweight and survival. We hypothesized that gestational binge alcohol exposure impairs maternal uterine vascular function, affecting endothelial nitric oxide (NO)-mediated vasodilation. Pregnant rats grouped as pair-fed control or binge alcohol exposed received a once-daily, orogastric gavage of isocaloric maltose-dextrin or alcohol, respectively. On gestational day 20, primary uterine arteries were isolated, cannulated, and connected to a pressure transducer, and functional studies were conducted by dual-chamber arteriography. Uterine arteries maintained at constant intramural pressure (90 mm Hg) were maximally constricted with thromboxane, and a dose-response for acetylcholine (Ach) was recorded. The alcohol group exhibited significantly impaired endothelium-dependent, Ach-induced uterine artery relaxation (↓∼30%). Subsequently, a dose-response was recorded following inhibition of endothelium-derived hyperpolarizing factor (apamin and TRAM-34) and prostacyclin (indomethacin). Ach-induced relaxation in the pair-fed control decreased by ~46%, and interestingly, relaxation in alcohol group further decreased by an additional ~48%, demonstrating that gestational binge alcohol impairs the NO system in the primary uterine artery. An endothelium-independent sodium nitroprusside effect was not observed. Immunoblotting indicated that alcohol decreased the level of endothelial excitatory P-Ser1177 endothelial NO synthase (eNOS) (p < 0.05) and total eNOS expression (p < 0.05) compared to both the normal and pair-fed controls. P-Ser1177 eNOS level was also confirmed by immunofluorescence imaging. This is the first study to demonstrate maternal binge alcohol consumption during pregnancy disrupts uterine artery vascular function via impairment of the eNOS vasodilatory system.


Anisotropic diffusion of membrane proteins at experimental timescales.
Single-particle tracking (SPT) experiments of lipids and membrane proteins provide a wealth of information about the properties of biomembranes. Careful analysis of SPT trajectories can reveal deviations from ideal Brownian behavior. Among others, this includes confinement effects and anomalous diffusion, which are manifestations of both the nanoscale structure of the underlying membrane and the structure of the diffuser. With the rapid increase in temporal and spatial resolution of experimental methods, a new aspect of the motion of the particle, namely, anisotropic diffusion, might become relevant. This aspect that so far received only little attention is the anisotropy of the diffusive motion and may soon provide an additional proxy to the structure and topology of biomembranes. Unfortunately, the theoretical framework for detecting and interpreting anisotropy effects is currently scattered and incomplete. Here, we provide a computational method to evaluate the degree of anisotropy directly from molecular dynamics simulations and also point out a way to compare the obtained results with those available from SPT experiments. In order to probe the effects of anisotropic diffusion, we performed coarse-grained molecular dynamics simulations of peripheral and integral membrane proteins in flat and curved bilayers. In agreement with the theoretical basis, our computational results indicate that anisotropy can persist up to the rotational relaxation time [τ=(2Dr)-1], after which isotropic diffusion is observed. Moreover, the underlying topology of the membrane bilayer can couple with the geometry of the particle, thus extending the spatiotemporal domain over which this type of motion can be detected.


Local Anesthetic Systemic Toxicity During Transversus Abdominis Plane Block With Liposomal Bupivacaine.
Local anesthetic systemic toxicity (LAST) is a rare but potentially fatal outcome associated with local anesthetic administration. Liposomal bupivacaine (LB; EXPAREL®) is a widely used local anesthetic with extended-release and liposomal formulation that carries an improved cardiac and central nervous system safety profile. However, there is limited data regarding LAST associated with liposomal bupivacaine. Here is described a case of local anesthetic systemic toxicity in a 68-year-old male who presented with obstructing sigmoid adenocarcinoma and underwent open sigmoidectomy with end descending colostomy. The operation was complicated by LAST following transversus abdominis plane block injection with liposomal bupivacaine resulting in cardiac arrest. Return of spontaneous circulation was achieved following advanced cardiac life support and infusion of 20% I.V. fat emulsion. Given the widespread use of local anesthetics, providers must be aware of the pathophysiology, diagnosis, and immediate treatment of LAST.


Merkel Cells Release Glutamate Following Mechanical Stimulation: Implication of Glutamate in the Merkel Cell-Neurite Complex.
Merkel cells (MCs) have been proposed to form a part of the MC-neurite complex with sensory neurons through synaptic contact. However, the detailed mechanisms for intercellular communication between MCs and neurons have yet to be clarified. The present study examined the increases in intracellular free Ca2+ concentration ([Ca2+]i) induced by direct mechanical stimulation of MCs. We also measured [Ca2+]i in the trigeminal ganglion neurons (TGs) following direct mechanical stimulation to the MCs in an MC-TGs coculture. The MCs were isolated from hamster buccal mucosa, while TGs were isolated from neonatal Wistar rats. Both cell populations showed depolarization-induced [Ca2+]i. Direct mechanical stimulation to MCs increased [Ca2+]i, showing stimulation strength dependence. In the MC-TGs coculture, the application of direct mechanical stimulation to MCs resulted in increased [Ca2+]i in the TGs. These changes were significantly suppressed by antagonists of glutamate-permeable anion channels (4,4'-diisothiocyanato-2,2'-stilbenedisulfonic acid; DIDS), and non-competitive antagonist of the N-methyl-D-aspartate (NMDA) receptors (MK801). Apyrase, an ATP-degrading enzyme, and suramin, a non-selective P2 purinergic receptor antagonist, did not exert inhibitory effects on these [Ca2+]i increases in the TGs following MC stimulation. These results indicated that MCs are capable of releasing glutamate, but not ATP, in response to cellular deformation by direct mechanical stimulation. The released glutamate activates the NMDA receptors on TGs. We suggest that MCs act as mechanoelectrical transducers and establish synaptic transmission with neurons, through the MC-neurite complex, to mediate mechanosensory transduction.


Comparison of methods for evaluating exercise-induced changes in thromboxane B2 and beta-thromboglobulin.
This study compared the effects of exercise or TXB2 and beta-TG when evaluated by four methods: 1) not adjusted; 2) adjusted for plasma volume changes (PV); 3) standardized per 10(5) platelets (PC); 4) or both PC and PV (PC-PV). Blood was collected from 16 men (41.3 +/- 8.1 yr) at rest after 30 min of exercise (IPE) and after 30 min recovery. Resting TXB2 and beta-TG concentrations were 62.0 +/- 6.2 pg.mL-1 and 129.8 +/- 12.5 ng.mL-1, respectively. When expressed on a per 10(5) platelet basis, resting PCTXB2 was 23.8 +/- 2.8 pg.mL-1.10(5-1) platelets and PC beta-TG was 50.77 +/- 6.0 ng.mL-1. 10(5-1) platelets. At IPE, TXB2 decreased 20.5% and beta-TG increased 13.6%. Thirty minutes after exercise TXB2 was 4.2% lower than resting values, whereas beta-TG was 26% higher. TXB2, beta-TG, PVTXB2, and PV beta-TG were not significantly altered by exercise. The only significant changes in TXB2 occurred at IPE when values were adjusted for changes in platelet count. At IPE, PCTXB2, and PC-PVTXB2 decreased 32.8% and 33.6%, respectively (P < 0.05). Similarly, beta-TG were not altered significantly by exercise except when the samples taken after 30 min of recovery were adjusted for changes in platelet count. At 30 min post-exercise PC beta-TG and PC-PV beta-TG were 21.2% and 28.4% greater (P < 0.05) than the resting beta-TG values. These data suggest that methods used to adjust concentrations of platelet derived substances for changes owing to exercise may influence conclusions about the effect of exercise on platelet function. Thus, it is imperative that researchers consider the purpose for which they are collecting TXB2 and beta-TG, as well as other constituents derived from blood cells, before they determine which methods of analysis to use.


Impact of major different variants of papillary thyroid microcarcinoma on the clinicopathological characteristics: the study of 1041 cases.
The incidence of papillary thyroid microcarcinoma (PTMC) has been increasing globally in the past few decades. PTMC does not have a distinctive morphology that results in differences in biological behavior. The aim of this study was to classify PTMCs according to the morphological features and explore the relationship with clinicopathological characteristics. Additionally, we sought to evaluate whether different variants of PTMC can be an independent predictor for lymph mode metastasis when considering other risk factors. Between December 2014 and December 2015, 1041 PTMC cases undergoing surgical resection at Tianjin Medical University Cancer Institute and Hospital were reviewed retrospectively. Statistical analysis was performed to investigate the independent factors for lymph node metastasis in PTMC. Conventional variant PTMC (CPTMC), follicular variant PTMC (FPTMC), and encapsulated variant PTMC (EnPTMC) were major variants in PTMC, collectively accounting for 96.7% of the entire PTMC cohort.There were significant differences in clinicopathological characteristics among the three major variants. The frequency of aggressive parameters was significantly different among the three variants, including tumor size, minimal extrathyroidal extension (minimal ETE), and lymph node metastasis (all P < 0.05), being highest in CPTMC, lowest in EnPTMC, and intermediate in FPTMC. FPTMC (OR = 0.642, P = 0.003) and EnPTMC (OR = 0.540, P = 0.041) were independent protective factors for lymph node metastasis (LNM). In contrast, male gender (OR = 1.836, P = 0.000), age less than 45 years (OR = 1.457, P = 0.009), tumor size greater than 0.5 cm (OR = 1.453, P = 0.007), calcification (OR = 1.465, P = 0.016), minimal ETE (OR = 1.801, P = 0.001), and multifocality (OR = 1.721, P = 0.000) were independent risk factors for LNM. The present study demonstrates the distinct biological behaviors of the three major PTMC variants and establishes an aggressive order of CPTMC ≫ FPTMC > EnPTMC. It is necessary to take into consideration variant-related risks and other independent predictors for the determination of lymphadenectomy in patients with PTMC.


A third major locus for autosomal dominant hypercholesterolemia maps to 1p34.1-p32.
Autosomal dominant hypercholesterolemia (ADH), one of the most frequent hereditary disorders, is characterized by an isolated elevation of LDL particles that leads to premature mortality from cardiovascular complications. It is generally assumed that mutations in the LDLR and APOB genes account for ADH. We identified one large French pedigree (HC2) and 12 additional white families with ADH in which we excluded linkage to the LDLR and APOB, implicating a new locus we named "FH3." A LOD score of 3.13 at a recombination fraction of 0 was obtained at markers D1S2892 and D1S2722. We localized the FH3 locus to a 9-cM interval at 1p34.1-p32. We tested four regional markers in another set of 12 ADH families. Positive LOD scores were obtained in three pedigrees, whereas linkage was excluded in the others. Heterogeneity tests indicated linkage to FH3 in approximately 27% of these non-LDLR/non-APOB ADH families and implied a fourth locus. Radiation hybrid mapping located four candidate genes at 1p34.1-p32, outside the critical region, showing no identity with FH3. Our results show that ADH is genetically more heterogeneous than conventionally accepted.


Mitosis inhibitors in anticancer therapy: When blocking the exit becomes a solution.
Current microtubule-targeting agents (MTAs) remain amongst the most important antimitotic drugs used against a broad range of malignancies. By perturbing spindle assembly, MTAs activate the spindle assembly checkpoint (SAC), which induces mitotic arrest and subsequent apoptosis. However, besides toxic side effects and resistance, mitotic slippage and failure in triggering apoptosis in various cancer cells are limiting factors of MTAs efficacy. Alternative strategies to target mitosis without affecting microtubules have, thus, led to the identification of small molecules, such as those that target spindle Kinesins, Aurora and Polo-like kinases. Unfortunately, these so-called second-generation of antimitotics, encompassing mitotic blockers and mitotic drivers, have failed in clinical trials. Our recent understanding regarding the mechanisms of cell death during a mitotic arrest pointed out apoptosis as the main variable, providing an opportunity to control the cell fates and influence the effectiveness of antimitotics. Here, we provide an overview on the second-generation of antimitotics, and discuss possible strategies that exploit SAC activity, mitotic slippage/exit and apoptosis induction, in order to improve the efficacy of anticancer strategies that target mitosis.


Transition metal-catalyzed site- and regio-divergent C-H bond functionalization.
Recent advances in transition metal-catalyzed C-H bond functionalization have profoundly impacted synthetic strategy. Since organic substrates typically contain several chemically distinct C-H bonds, controlling the regioselectivity of C-H bond functionalization is imperative to harness its full potential. Moreover, the ability to alter reaction pathways to selectively functionalize different C-H bonds in a substrate represents a greater opportunity and challenge. The choice of catalysts, ligands, solvents, and even more subtle variations of the reaction conditions have been shown to allow the formation of regioisomeric C-H functionalization products starting from the same precursors. This review describes recent advances in transition metal-catalyzed divergent C-H bond functionalization that highlight its potential in organic synthesis.


Therapeutic Switching of Rafoxanide: a New Approach To Fighting Drug-Resistant Bacteria and Fungi.
Control and management of life-threatening bacterial and fungal infections are a global health challenge. Despite advances in antimicrobial therapies, treatment failures for resistant bacterial and fungal infections continue to increase. We aimed to repurpose the anthelmintic drug rafoxanide for use with existing therapeutic drugs to increase the possibility of better managing infection and decrease treatment failures. For this purpose, we evaluated the antibacterial and antifungal potential of rafoxanide. Notably, 70% (70/100) of bacterial isolates showed multidrug resistance (MDR) patterns, with higher prevalence among human isolates (73.5% [50/68]) than animal ones (62.5% [20/32]). Moreover, 22 fungal isolates (88%) were MDR and were more prevalent among animal (88.9%) than human (87.5%) sources. We observed alarming MDR patterns among bacterial isolates, i.e., Klebsiella pneumoniae (75% [30/40; 8 animal and 22 human]) and Escherichia coli (66% [40/60; 12 animal and 28 human]), and fungal isolates, i.e., Candida albicans (86.7% [13/15; 4 animal and 9 human]) and Aspergillus fumigatus (90% [9/10; 4 animal and 5 human]), that were resistant to at least one agent in three or more different antimicrobial classes. Rafoxanide had antibacterial and antifungal activities, with minimal inhibitory concentration (MICs) ranging from 2 to 128 μg/mL. Rafoxanide at sub-MICs downregulated the mRNA expression of resistance genes, including E. coli and K. pneumoniae blaCTX-M-1, blaTEM-1, blaSHV, MOX, and DHA, C. albicans ERG11, and A. fumigatus cyp51A. We noted the improvement in the activity of β-lactam and antifungal drugs upon combination with rafoxanide. This was apparent in the reduction in the MICs of cefotaxime and fluconazole when these drugs were combined with sub-MIC levels of rafoxanide. There was obvious synergism between rafoxanide and cefotaxime against all E. coli and K. pneumoniae isolates (fractional inhibitory concentration index [FICI] values ≤ 0.5). Accordingly, there was a shift in the patterns of resistance of 16.7% of E. coli and 22.5% of K. pneumoniae isolates to cefotaxime and those of 63.2% of C. albicans and A. fumigatus isolates to fluconazole when the isolates were treated with sub-MICs of rafoxanide. These results were confirmed by in silico and mouse protection assays. Based on the in silico study, one possible explanation for how rafoxanide reduced bacterial resistance is through its inhibitory effects on bacterial and fungal histidine kinase enzymes. In short, rafoxanide exhibited promising results in overcoming bacterial and fungal drug resistance. IMPORTANCE The drug repurposing strategy is an alternative approach to reducing drug development timelines with low cost, especially during outbreaks of disease caused by drug-resistant pathogens. Rafoxanide can disrupt the abilities of bacterial and fungal cells to adapt to stress conditions. The coadministration of antibiotics with rafoxanide can prevent the failure of treatment of both resistant bacteria and fungi, as the resistant pathogens could be made sensitive upon treatment with rafoxanide. From our findings, we anticipate that pharmaceutical companies will be able to utilize new combinations against resistant pathogens.


Combined sialic acid and histone deacetylase (HDAC) inhibitor treatment up-regulates the neuroblastoma antigen GD2.
Neuroblastoma cells highly express the disialoganglioside GD2, a tumor-associated carbohydrate antigen, which is only sparsely expressed on healthy tissue. GD2 is a primary target for the development of immunotherapy for neuroblastoma. Immunotherapy with monoclonal anti-GD2 antibodies has proven safety and efficacy in clinical trials and is included in the standard treatment for children with high-risk neuroblastoma. Strategies to modulate GD2 expression in neuroblastoma could further improve anti-GD2-targeted immunotherapy. Here, we report that the cellular sialylation pathway, as well as epigenetic reprogramming, strongly modulates GD2 expression in human and mouse neuroblastoma cell lines. Recognition of GD2 by the 14G2a antibody is sialic acid-dependent and was blocked with the fluorinated sialic acid mimetic Ac53FaxNeu5Ac. Interestingly, sialic acid supplementation using a cell-permeable sialic acid analogue (Ac5Neu5Ac) boosted GD2 expression without or with minor alterations in overall cell surface sialylation. Furthermore, sialic acid supplementation with Ac5Neu5Ac combined with various histone deacetylase (HDAC) inhibitors, including vorinostat, enhanced GD2 expression in neuroblastoma cells beyond their individual effects. Mechanistic studies revealed that Ac5Neu5Ac supplementation increased intracellular CMP-Neu5Ac concentrations, thereby providing higher substrate levels for sialyltransferases. Furthermore, HDAC inhibitor treatment increased mRNA expression of the sialyltransferases GM3 synthase (ST3GAL5) and GD3 synthase (ST8SIA1), both of which are involved in GD2 biosynthesis. Our findings reveal that sialic acid analogues and HDAC inhibitors enhance GD2 expression and could potentially be employed to boost anti-GD2 targeted immunotherapy in neuroblastoma patients.


Manganese-calcium clusters supported by calixarenes.
The structure of the oxygen-evolving complex of photosystem II, which contains a cubane-like metal-oxo cluster incorporating four manganese(III,IV) cations, along with a calcium cation, has focussed attention on synthetic analogues of this cluster. Despite this activity, there are relatively few structurally characterised coordination clusters with this combination of metal cations. The calixarenes are synthetically versatile and well established cluster-supporting ligands, which to date have not been reported to support a calcium/manganese cluster. Here we report that p-t-butylthiacalix[4]arene supports CaMn2 and Ca2Mn2 clusters, whereas reactions of p-t-butylcalix[4]arene, p-t-butylsulfinylcalix[4]arene, and p-t-butylsulfonylcalix[4]arene, under the same conditions, produced only homometallic manganese complexes.


Sight and blindness in the same person: Gating in the visual system.
We present the case of a patient having dissociative identity disorder (DID) who-after 15 years of misdiagnosed cortical blindness--step-by-step regained sight during psychotherapeutic treatment. At first only a few personality states regained vision whereas others remained blind. This could be confirmed by electrophysiological measurement, in which visual evoked potentials (VEPs) were absent in the blind personality states but were normal and stable in the seeing states. A switch between these states could happen within seconds. We assume a top-down modulation of activity in the primary visual pathway as a neural basis of such psychogenic blindness, possibly at the level of the thalamus. VEPs therefore do not allow separating psychogenic blindness from organic disruption of the visual pathway. In summary, psychogenic blindness seems to suppress visual information at an early neural stage.


Recent Advances in Negative Capacitance FinFETs for Low-Power Applications: A Review.
In the contemporary era of Internet-of-Things (IoT), there is an extensive search for competent devices which can operate at ultralow voltage supply. Due to the restriction of power dissipation, a reduced sub-threshold swing-based device appears to be the perfect solution for efficient computation. To counteract this issue, negative capacitance fin field-effect transistors (NC-FinFETs) came up as the next-generation platform to withstand the aggressive scaling of transistors. The ease of fabrication, process-integration, higher current driving capability, and ability to tailor the short-channel effects (SCEs) are some of the potential advantages offered by NC-FinFETs that attracted the attention of researchers worldwide. The following review emphasizes how this new state-of-art technology supports the persistence of Moore's law and addresses the ultimate limitation of Boltzmann tyranny by offering a sub-threshold slope (SS) below 60 mV/decade. The article primarily focuses on two parts: 1) the theoretical background of negative capacitance (NC) effect and FinFET devices and 2) the recent progress done in the field of NC-FinFETs. It also highlights the crucial areas that need to be upgraded, to mitigate the challenges faced by this technology and the future prospects of such devices.


Cardiac Arrest Survival in Pediatric and General Emergency Departments.
Pediatric out-of-hospital cardiac arrest (OHCA) has a low rate of survival to hospital discharge. Understanding whether pediatric emergency departments (EDs) have higher survival than general EDs may help identify ways to improve care for all patients with OHCA. We sought to determine if OHCA survival differs between pediatric and general EDs. We used the 2009-2014 Nationwide Emergency Department Sample to study children under 18 with cardiac arrest. We compared pediatric EDs (those with >75% pediatric visits) to general EDs on the outcome of survival to hospital discharge or transfer. We determined unadjusted and adjusted survival, accounting for age, region, and injury severity. Analyses were stratified by nontraumatic versus traumatic cause. The incidences of nontraumatic and traumatic OHCA were 7.91 (95% confidence interval [CI]: 7.52-8.30) and 2.67 (95% CI: 2.49-2.85) per 100 000 person years. In nontraumatic OHCA, unadjusted survival was higher in pediatric EDs than general EDs (33.8% vs 18.9%, P < .001). The adjusted odds ratio of survival in pediatric versus general EDs was 2.2 (95% CI: 1.7-2.8). Children with traumatic OHCA had similar survival in pediatric and general EDs (31.7% vs 26.1%, P = .14; adjusted odds ratio = 1.3 [95% CI: 0.8-2.1]). In a nationally representative sample, survival from nontraumatic OHCA was higher in pediatric EDs than general EDs. Survival did not differ in traumatic OHCA. Identifying the features of pediatric ED OHCA care leading to higher survival could be translated into improved survival for children nationally.


Spatiotemporal analysis of purkinje cell degeneration relative to parasagittal expression domains in a model of neonatal viral infection.
Infection of newborn Lewis rats with Borna disease virus (neonatal Borna disease [NBD]) results in cerebellar damage without the cellular inflammation associated with infections in later life. Purkinje cell (PC) damage has been reported for several models of early-life viral infection, including NBD; however, the time course and distribution of PC pathology have not been investigated rigorously. This study examined the spatiotemporal relationship between PC death and zonal organization in NBD cerebella. Real-time PCR at postnatal day 28 (PND28) revealed decreased cerebellar levels of mRNAs encoding the glycolytic enzymes aldolase C (AldoC, also known as zebrin II) and phosphofructokinase C and the excitatory amino acid transporter 4 (EAAT4). Zebrin II and EAAT4 immunofluorescence analysis in PND21, PND28, PND42, and PND84 NBD rat cerebella revealed a complex pattern of PC degeneration. Early cell loss (PND28) was characterized by preferential apoptotic loss of zebrin II/EAAT4-negative PC subsets in the anterior vermis. Consistent with early preferential loss of zebrin II/EAAT4-negative PCs in the vermis, the densities of microglia and the Bergmann glial expression of metallothionein I/II and the hyaluronan receptor CD44 were higher in zebrin II/EAAT4-negative zones. In contrast, early loss in lateral cerebellar lobules did not reflect a similar discrimination between PC phenotypes. Patterns of vermal PC loss became more heterogeneous at PND42, with the loss of both zebrin II/EAAT4-negative and zebrin II/EAAT4-positive neurons. At PND84, zebrin II/EAAT4 patterning was abolished in the anterior cerebellum, with preferential PC survival in lobule X. Our investigation reveals regional discrimination between patterns of PC subset loss, defined by zebrin II/EAAT4 expression domains, following neonatal viral infection. These findings suggest a differential vulnerability of PC subsets during the early stages of virus-induced neurodegeneration.


Iterative Rectifying Methods for Nonrepetitive Continuous-Time Learning Control Systems.
To implement iterative learning control (ILC), one of the most fundamental hypotheses is the strict repetitiveness (i.e., iteration-independence) of the controlled systems, especially of their plant models. This hypothesis, however, results in difficulties of developing theoretic analysis methods and promoting practical applications for ILC, especially in the presence of continuous-time systems, which is the motivation of the current paper to cope with robust tracking problems of continuous-time ILC systems subject to nonrepetitive (i.e., iteration-dependent) uncertainties. Based on integrating an iterative rectifying mechanism, continuous-time ILC can effectively address the ill effects of the multiple nonrepetitive uncertainties that arise from the system models, initial states, load and measurement disturbances, and desired references. Furthermore, a robust convergence analysis method is presented for continuous-time ILC by combining a contraction mapping-based method and a system equivalence transformation method. It is disclosed that regardless of continuous-time ILC systems with zero or nonzero system relative degrees, the robust tracking tasks in the presence of nonrepetitive uncertainties can be accomplished, together with the boundedness of all the system trajectories being ensured. Two examples are included to verify the validity of our robust tracking results for nonrepetitive continuous-time ILC systems.


A method to determine the presence of averaged event-related fields using randomization tests.
We present a simple and effective method to test whether an event consistently activates a set of brain electric sources across repeated measurements of event-related scalp field data. These repeated measurements can be single trials, single subject ERPs, or ERPs from different studies. The method considers all sensors simultaneously, but can be applied separately to each time frame or frequency band of the data. This allows limiting the analysis to time periods and frequency bands where there is positive evidence of a consistent relation between the event and some brain electric sources. The test may therefore avoid false conclusions about the data resulting from an inadequate selection of the analysis window and bandpass filter, and permit the exploration of alternate hypotheses when group/condition differences are observed in evoked field data. The test will be called topographic consistency test (TCT). The statistical inference is based on simple randomization techniques. Apart form the methodological introduction, the paper contains a series of simulations testing the statistical power of the method as function of number of sensors and observations, a sample analysis of EEG potentials related to self-initiated finger movements, and Matlab source code to facilitate the implementation. Furthermore a series of measures to control for multiple testing are introduced and applied to the sample data.


The cytologic diagnosis of hydatid disease.
Eleven cases of hydatid disease diagnosed by cytologic methods are reported. sputum specimens and aspirated peritoneal material were embedded in paraffin and then processed by conventional procedures. Smears were obtained from pleural and ascitic fluids. Sputum cytology gave 33% positive results. The total rate of positive cytology in all echinococosis cases, including sputum, peritoneal material and pleural fluids, was 40%. Silver methenamine and Best carmine procedures were of special interest for the identification of laminated layer fragments.


Inhibitory effect of CUL1 on atherosclerosis through the p53 pathway.
Atherosclerosis (AS) is a serious chronic condition associated with cardiovascular and cerebrovascular diseases. Research on AS is currently lacking, and there is a need to increase research to improve the diagnosis, treatment, and prognosis of AS. Therefore, the aim of the present study was to explore the molecular mechanism and identify potential biomarkers in AS. First, we downloaded the GSE28829 dataset and screened differentially expressed genes (DEGs). Then, DEGs were analyzed by functional enrichment analysis and protein-protein interaction (PPI) networks to determine hub genes. The key gene for AS was identified following the expression analysis of AS, clinical correlation with immune factors, the gene set enrichment analysis (GSEA) enrichment pathway, and receiver-operating characteristic curve analysis. In functional experiments, correlations between cullin 1 (CUL1), cytokines, and downstream targets in AS were investigated. We identified 595 upregulated and 391 downregulated DEGs enriched in neutrophil degranulation, the B-cell receptor signaling pathway, cell-matrix adhesion, and fatty acid degradation. Through PPI, we identified 7 hub genes for the expression analysis, immunoassay, and GSEA. Finally, CUL1 was identified as the inhibitory gene in AS associated with immune factors, and was found to have a strong prognostic prediction ability. The results indicated that CUL1 upregulated interleukin (IL)-6, IL-1β, and tumor necrosis factor-α concentrations, and weakened the cell proliferation of AS. It was also found that CUL1 exerted its inhibitory function in AS by the p53 pathway. The findings of the present study indicate that CUL1 is a suppressing gene in AS, and has the potential to be a therapeutic and prognostic biomarker for AS.


Acute Wilson's disease and thalassemia minor: a case report.
The case of a 17-year-old Italian girl with thalassemia minor and fatal acute Wilson's disease together with severe hemolysis is reported. The fortuitous(?) occurrence of both genetically determined conditions in the same patient was the source of delayed diagnosis of the metabolic disorder.


The use of amlodipine, but not of P-glycoprotein inhibiting calcium channel blockers is associated with clopidogrel poor-response.
Clopidogrel is a prodrug that has to be converted in vivo to its active metabolite by cytochrome (CYP)P450 iso-enzymes. As calcium channel blockers (CCBs) are inhibitors of CYP3A4, concomitant use of these drugs might play a role in the wide inter-individual variability in the response to clopidogrel. However, some CCBs also have strong inhibitory effects on the drug transporter P-glycoprotein (Pgp), which mediates clopidogrel's intestinal absorption. It was the aim of this study to evaluate the effect of co-administration of Pgp-inhibiting and non-Pgp-inhibiting CCBs on on-clopidogrel platelet reactivity in patients on dual antiplatelet therapy undergoing elective percutaneous coronary intervention (PCI). In a total of 623 consecutive patients undergoing elective PCI treated with clopidogrel and aspirin, platelet reactivity to 5 and 20 muM adenosine diphospate (ADP) and clopidogrel poor-response (defined as > 70% platelet aggregation to 20 muM ADP) were evaluated by light transmittance aggregometry. A total of 222 patients (35.6%) were on CCB treatment, of which 98 used Pgp-inhibiting CCBs (verapamil, nifedipine, diltiazem, barnidipine) and 124 patients used the non-Pgp-inhibiting CCB amlodipine. Adjusted mean ADP-induced on-clopidogrel platelet reactivity was significantly higher in both users of Pgp-inhibiting CCBs and amlodipine as compared to CCB non-users (all p<0.05). However, only the use of amlodipine was significantly associated with a 2.3-fold increased risk of clopidogrel poor-response. This study demonstrates that concomitant use of Pgp-inhibiting CCBs and amlodipine increases on-clopidogrel platelet reactivity. Only amlodipine was associated with clopidogrel poor-response. The drug-drug interaction between clopidogrel and amlodipine might be more clinically relevant as compared to P-glycoprotein-inhibiting CCBs.


Coexistence of deficiencies of uroporphyrinogen III synthase and decarboxylase in a patient with congenital erythropoietic porphyria and in his family.
A hitherto undescribed dual deficiency of uroporphyrinogen III synthase and uroporphyrinogen decarboxylase was observed in the erythrocytes in a 14 year-old patient who had presented with congenital erythropoietic porphyria since early childhood. Whereas congenital erythropoietic porphyria was metabolically and clinically overt, a hereditary deficiency of uroporphyrinogen decarboxylase was confirmed by family study. The uroporphyrinogen III synthase activity of the propositus was decreased to 26% of the control while his asymptomatic family members had activities between 53-65% of the control. Additionally, the uroporphyrinogen decarboxylase activity was 55-66% of the control in the patient and his family. Family investigations have shown that the two disorders do not consistently segregate together. Although urinary porphyrin excretions of relatives were in the physiological range, the proportion of coproporphyrin isomer I showed a relative increase, which can serve as a biochemical indicator for heterozygous uroporphyrinogen III synthase gene carriers.


Early trajectory features and the course of psychotherapy.
Objective: We investigated the relationship between psychotherapy outcome and trajectory variables measured during the first five treatment sessions. Method: In a psychotherapy training clinic, 158 patients completed a measure of symptomatic distress at each visit during treatment lasting a median 30 sessions. We investigated outcome predictions based on symptomatic distress at intake, change in distress between the first and fifth sessions, and intersession shifts. Results: Elevated symptomatic distress at intake predicted reliable change, while limited initial distress predicted clinically significant improvement; both predictions were independent of those by other trajectory variables. Large early change and large intersession shifts yielded similar predictions of reliable change and clinically significant improvement. Greater change between the first and fifth sessions predicted less change in subsequent sessions. By contrast, neither initial distress nor early intersession shifts predicted change in distress after the fifth session. Conclusion: Considered together, symptomatic distress at intake and change in distress over the first five sessions improve the prediction of psychotherapy outcome over base rate. Intersession shifts contribute little to the prediction of outcome after accounting for overall change in symptomatic distress during the first five sessions.


TrkC: a new predictive marker in breast cancer?
Available data involve neurotrophins and their receptors in carcinomas. Quantitative evaluation of these molecules in these tumors might be useful as prognostic marker and eventual treatments. Our study on 40 mammary tumors tries to correlate expression of these molecules and prognosis. Immunohistochemistry for NGF, BDNF, NT3, TrkA, TrkB, TrkC, and p75 was used. Patient's age, histopathology, Bloom-Richardson grading, estrogen and progesterone receptors, Ki-67 index, HER-2, p53 were evaluated. Statistics found inverse relationship between grading and TrkC expression. We found significantly higher TrkC expression in Grade I than in Grade III tumors. Rise in TrkC expression could indicate good prognosis.


A New Horizon in Vitamin K Research.
Vitamin K is a cofactor for γ-glutamyl carboxylase, which catalyzes the posttranslational conversion of specific glutamyl residues to γ-carboxyglutamyl residues in a variety of vitamin K-dependent proteins (VKDPs) involved in blood coagulation, bone and cartilage metabolism, signal transduction, and cell proliferation. Despite the great advances in the genetic, structural, and functional studies of VKDPs as well as the enzymes identified as part of the vitamin K cycle which enable it to be repeatedly recycled within the cells, little is known of the identity and roles of key regulators of vitamin K metabolism in mammals and humans. This review focuses on new insights into the molecular mechanisms underlying the intestinal absorption and in vivo tissue conversion of vitamin K1 to menaquinone-4 (MK-4) with special emphasis on two major advances in the studies of intestinal vitamin K transporters in enterocytes and a tissue MK-4 biosynthetic enzyme UbiA prenyltransferase domain-containing protein 1 (UBIAD1), which participates in the in vivo conversion of a fraction of dietary vitamin K1 to MK-4 in mammals and humans, although it remains uncertain whether UBIAD1 functions as a key regulator of intracellular cholesterol metabolism, bladder and prostate tumor cell progression, vascular integrity, and protection from oxidative stress.


Incidental rectal mucosa obtained via transrectal ultrasound-guided prostatic core biopsies.
A retrospective review was conducted of material from 782 transrectal ultrasound-guided prostatic core biopsies to determine whether incidental pieces of rectal mucosa obtained in this manner could harbor clinically significant rectal pathology or incur artifacts that cause diagnostic difficulty. Material from 114 biopsies (14.6%) showed rectal mucosa, and material from 19 (16.7%) showed rectal pathology, including a hyper-plastic polyp in 1, changes consistent with ulcerative proctitis in 8, focal active proctitis in 7, and granulomas in 3. The original pathologist overlooked the hyperplastic polyp. In 1 specimen, rectal lymphocytes and plasma cells that were displaced over prostatic tissue closely mimicked prostatic adenocarcinoma (Gleason score 5). Conversely, in another specimen, prostatic adenocarcinoma (Gleason score 5) that was displaced near rectal mucosa closely mimicked a rectal lymphoid aggregate. Incidental rectal mucosa obtained via transrectal ultrasound-guided prostatic core biopsies occasionally harbors clinically significant rectal pathology and rarely incurs artifacts that cause diagnostic difficulty.


Preemptive erythropoietin plus high ribavirin doses to increase rapid virological responses in HIV patients treated for chronic hepatitis C.
Chronic hepatitis C affects one-third of HIV(+) patients worldwide. High ribavirin (RBV) exposure is crucial to maximize the response to hepatitis C therapy in this population, although it may increase the risk for hemolytic anemia. PERICO is a prospective multicenter trial in which HIV/HCV-coinfected patients are randomized to receive peginterferon (pegIFN) alfa-2a 180 microg/week plus either weight-based RBV (1000-1200 mg/day) or RBV 2000 mg/day, the latest along with erythropoietin alfa (EPO) 30,000 IU/week from the first day until week 4. A total of 149 patients were assessed in a planned interim analysis at week 4. In both arms, 22% of patients achieved negative HCV-RNA (rapid virological response, RVR). Multivariate analysis [OR (IC 95%), p] showed that factors associated with RVR were HCV genotypes 2/3 vs. 1/4 [20 (5-100), <0.01] and baseline HCV-RNA [0.16 (0.07-0.37) per log IU/ml, <0.01]. The occurrence of severe anemia (hemoglobin <10 g/dl) did not differ when comparing RBV vs. high RBV + EPO (7% vs. 3%; p = 0.4). Moreover, RBV plasma trough levels were comparable at week 4 (1.9 vs. 2.4 microg/ml; p = 0.2). Use of high RBV doses with preemptive EPO during the first 4 weeks of hepatitis C therapy is safe, but fails to enhance significantly RBV plasma exposure and RVR rates. Extensive intraerythrocyte accumulation of RBV following boosted production of red blood cells by EPO could explain these findings.


Cathodic Aromatic C,C Cross-Coupling Reaction via Single Electron Transfer Pathway.
We have successfully developed a novel cathodic cross-coupling reaction of aryl halides with arenes. Utilization of the cathodic single electron transfer (SET) mechanism for activation of aryl halides enables the cross-coupling reaction to proceed without the need for any transition metal catalysts or single electron donors in a mild condition. The SET from a cathode to an aryl halide initiates a radical chain by giving an anion radical of the aryl halide. The following propagation cycle also consists entirely of anion radical intermediates.


Activation of p38 mitogen-activated protein kinase in spinal cord contributes to chronic constriction injury-induced neuropathic pain.
The present study aimed to investigate the role of spinal p38 mitogen-activated protein kinase (p38 MAPK) activation in chronic constriction injury (CCI) of the sciatic nerve induced neuropathic pain. CCI model was produced by loosely ligating the left sciatic nerve proximal to the sciatica's trifurcation with 4-0 silk thread in male Sprague-Dawley rat. SB203580, a specific inhibitor of the p38 MAPK, was intrathecally administered on day 5 post-CCI. Thermal and mechanical nociceptive thresholds were assessed with the paw withdrawal lantency (PWL) to radiant heat and the paw withdrawal threshold (PWT) to von Frey filaments respectively. The protein levels of the phosphorylated p38 MAPK (p-p38 MAPK) and phosphorylated cAMP response element binding protein (pCREB) were assessed by Western blot analysis. The results showed that CCI significantly increased the expressions of cytosolic and nuclear p-p38 MAPK in the spinal cord. Intrathecal administration of SB203580 dose-dependently reversed the established mechanical allodynia and thermal hyperalgesia induced by CCI. Correlated with behavior results, SB203580 dose-dependently inhibited the CCI-induced increase of the expressions of cytosolic and nuclear p-p38 MAPK and nuclear pCREB in the spinal cord. Taken together, these findings suggest that the activation of p38 MAPK pathway contributes to the development of neuropathic pain induced by CCI, and that the function of p-p38 MAPK may partly be accomplished via the CREB-dependent gene expression.


Anatomic Assessment of K-Wire Trajectory for Transverse Percutaneous Fixation of Small Finger Metacarpal Fractures: A Cadaveric Study.
The purpose of this cadaveric study is to evaluate the trajectory of percutaneous transverse Kirschner wire (K-wire) placement for fifth metacarpal fractures relative to the sagittal profile of the fifth metacarpal in order to develop a targeting strategy for the treatment of fifth metacarpal fractures. Using 12 unmatched fresh human upper limbs, we evaluated the trajectory of percutaneous transverse K-wire placement relative to the sagittal profile of the fifth metacarpal in order to develop a targeting strategy for treatment of fifth metacarpal fractures. The midpoint of the small and ring finger metacarpals in the sagittal plane was identified at 3 points. At each point, a K-wire was inserted from the small finger metacarpal into the midpoint of the ring finger metacarpal ("center-center" position). The angle of the transverse K-wire relative to the table needed to achieve a center-center position averaged 20.8°, 18.9°, and 16.7° for the proximal diaphysis, middiaphysis, and the collateral recess, respectively. Approximately 80% of transversely placed K-wires obtained purchase in the long finger metacarpal. These results can serve as a guide to help surgeons in the accurate placement of percutaneous K-wires for small finger metacarpal fractures and may aid in surgeon training.


Na/Ca exchanger overexpression induces endoplasmic reticulum-related apoptosis and caspase-12 activation in insulin-releasing BRIN-BD11 cells.
Ca(2+) may trigger programmed cell death (apoptosis) and regulate death-specific enzymes. Therefore, the development of strategies to control Ca(2+) homeostasis may represent a potential approach to prevent or enhance cell apoptosis. To test this hypothesis, the plasma membrane Na/Ca exchanger (NCX1.7 isoform) was stably overexpressed in insulin-secreting tumoral cells. NCX1.7 overexpression increased apoptosis induced by endoplasmic reticulum (ER) Ca(2+)-ATPase inhibitors, but not by agents increasing intracellular calcium concentration ([Ca(2+)](i)), through the opening of plasma membrane Ca(2+)-channels. NCX1.7 overexpression reduced the rise in [Ca(2+)](i) induced by all agents, depleted ER Ca(2+) stores, sensitized the cells to Ca(2+)-independent proapoptotic signaling pathways, and reduced cell proliferation by approximately 40%. ER Ca(2+) stores depletion was accompanied by the activation of the ER-specific caspase (caspase-12), and the activation was enhanced by ER Ca(2+)-ATPase inhibitors. Hence, Na/Ca exchanger overexpression, by depleting ER Ca(2+) stores, triggers the activation of caspase-12 and increases apoptotic cell death. By increasing apoptosis and decreasing cell proliferation, overexpression of Na/Ca exchanger may represent a new potential approach in cancer gene therapy. On the other hand, our results open the way to the development of new strategies to control cellular Ca(2+) homeostasis that could, on the contrary, prevent the process of apoptosis that mediates, in part, beta-cell autoimmune destruction in type 1 diabetes.


DIVCLUS: an automatic method in the GEANFAMMER package that finds homologous domains in single- and multi-domain proteins.
Large-scale determination of relationships between the proteins produced by genome sequences is now common. All protein sequences are matched and those that have high match scores are clustered into families. In cases where the proteins are built of several domains or duplication modules, this can lead to misleading results. Consider the very simple example of three proteins: 1, formed by duplication modules A and B; 2, formed by duplication modules B' and C; and 3, formed by duplication modules C' and D. Duplication modules B and B' are homologous, as are C and C'. Matching the sequences of 1, 2 and 3 followed by simple single-linkage clustering would put all three in the same family, even though proteins 1 and 3 are not related. This is because the different parts of 2 match 1 and 3. This paper describes a procedure, DIVCLUS, that divides such complex clusters of partially related sequences into simple clusters that contain only related duplication modules. In the example just given, it would produce two groups of sequences: the first with domains B of sequence 1 and B of sequence 2, and the second with domain C of sequence 2 and C of sequence 3. DIVCLUS is part of a package called GEANFAMMER, for GEnome ANalysis and protein FAMily MakER. The package automates the detection of families of duplication modules from a protein sequence database. DIVCLUS has been applied to the division of single-linkage clusters generated from the protein sequences of six completely sequenced bacterial genomes. Out of 12 013 genes in these six genomes, 4563 single- and multi-domain sequences formed 1071 complex clusters. Application of the DIVCLUS program resolved these clusters into 2113 clusters corresponding to single duplication modules. The perl5 program and its documentation are available at the following address: http://www.mrc-lmb.cam.ac.uk/genomes/ and by anonymous ftp at ftp.mrc-lmb.cam.ac.uk in the directory /pub/genomes/Software/. email@example.com; jong@mrc-lmb. cam.ac.uk


Influence of the status of the renin-angiotensin system on the effect of cilazapril on neointima formation after vascular injury in rats.
Angiotensin converting enzyme (ACE) inhibition has been shown to prevent neointima formation after vascular injury in rats. However, clinical results evaluating restenosis after angioplasty have been negative. The goal of the present study was to evaluate the influence of the renin-angiotensin system (RAS) status on the effect of ACE inhibition on neointima formation. Arterial injury was produced by ballooning the left carotid artery of rats, and neointima formation was evaluated by morphometry 2 weeks after ballooning. The effects of cilazapril were assessed in four experimental groups: normotensive rats, spontaneously hypertensive rats, hypertensive rats with a renal artery stenosis induced by clipping (two-kidney, one-clip rats), and hypertensive rats with uninephrectomy, high salt intake, and administration of deoxycorticosterone (DOCA). In parallel groups of rats, measurement of plasma renin activity was made in order to characterize (at least at the plasma level) the status of the RAS. As expected, renal artery stenosis markedly increased plasma renin activity, and DOCA decreased it to undetectable levels. Cilazapril had a marked preventive effect on neointima formation in normotensive rats, spontaneously hypertensive rats, and two-kidney, one-clip rats but was ineffective in DOCA rats. We conclude that the status of the RAS has a major influence on the effect of cilazapril on neointima formation after vascular injury.


[Occupational exposure in orthopedic procedures under fluoroscopic control].
In interventional radiology the highest radiation doses are usually recorded for both the medical staff and the patients. Interventional procedures with X-rays are implemented in a number of medical specializations. This paper concerns the exposure of interventional teams performing orthopedic procedures under X-rays control. Doses for interventional teams were measured in the 3 Łódź hospitals. Thermoluminescent dosemeters were applied to measure the following dose equivalents: Hp(3) for eye lens, Hp(0.07) for palm skin, Hp(10) at the level of the neck without a protective shield (i.e., collar) and Hp(10) for the whole body on the front surface of the trunk (measured under the protective apron at the level of the chest). Doses for the operator who performs surgery, assisting physicians and scrub nurse were measured during 95 procedures. The highest doses were received by the operator the dose for eyes per 1 procedure did not exceed 0.1 mSv, the highest dose for hands was 1.6 mSv and the highest recorded effective dose was 0.02 mSv. On the basis of the results of measurements and their comparison with the values reported in the literature it may be concluded that the obtained results fall within the published reference range (for non-vascular procedures). This proves the compatibility of practice in the monitored Łódź hospitals with routine methodology applied in other interventional departments. The measurement results confirm that the usage of thermoluminescent dosimetry is fully adequate for the evaluation of exposure in interventional radiology and that the usage of at least 2 dosemeters for that staff is necessary. Med Pr 2017;68(2):221-227.


Proposed mechanism for the formation of cold dense structures in plasmas.
The formation of cold dense structures in a hydrogen isotope plasma, caused by the penetration of impurities from a localized source, is modelled numerically. It is shown that a bubble structure with very high densities and low temperatures of all plasma components, i.e. electrons, main and impurity ions, arises if the density of impurity neutrals in the source exceeds a critical level. The major cause of this condensation phenomenon are Coulomb collisions between main and impurity ion species which, on the one hand, cool down the main ions and, on the other hand, confine impurity ions near the source. This mechanism is efficient even if the impurity radiation, considered usually as the most important cause of condensation instabilities, is completely switched off in the calculations. On the contrary, without ion-ion Coulomb collisions the radiation losses result only in a transient cooling of electrons with a later recovery of their temperature to a level comparable with the initial one.


[Mechanical emergency stimulation in asystole and extreme bradycardia].
Repetitive precordial thumping is the simplest method of temporary cardiac pacing. In 90 patients out of 100 with witnessed cardiac arrest because of asystole or marked bradycardia the critical situation could be effectively bridged over by this until a sufficient spontaneous rhythm returned or electrostimulation was ready to function. 69 patients were conscious during the stimulation. During the mechanical pacing only in 2 patients there occurred ventricular flutter or fibrillation, which was stopped by electric defibrillation. The existence of the myocardial contractility is the presupposition for effective mechanical and electrical pacing.


Comparing RNA secondary structures using a relaxed base-pair score.
The use of free energy-based algorithms to compute RNA secondary structures produces, in general, large numbers of foldings. Recent research has addressed the problem of grouping structures into a small number of clusters and computing a representative folding for each cluster. At the heart of this problem is the need to compute a quantity that measures the difference between pairs of foldings. We introduce a new concept, the relaxed base-pair (RBP) score, designed to give a more biologically realistic measure of the difference between structures than the base-pair (BP) metric, which simply counts the number of base pairs in one structure but not the other. The degree of relaxation is determined by a single relaxation parameter, t. When t = 0, (no relaxation) our method is the same as the BP metric. At the other extreme, a very large value of t will give a distance of 0 for identical structures and 1 for structures that differ. Scores can be recomputed with different values of t, at virtually no extra computation cost, to yield satisfactory results. Our results indicate that relaxed measures give more stable and more meaningful clusters than the BP metric. We also use the RBP score to compute representative foldings for each cluster.


Predictive value of F-18 FDG PET/CT quantization parameters in diffuse large B cell lymphoma: a meta-analysis with 702 participants.
F-18 fluorodeoxyglucose (FDG) positron emission tomography/computerized tomography (PET/CT) is considered to be the most beneficial imaging method for staging patients with lymphoma. Whether maximum standardized uptake value (SUVmax), metabolic tumor volume (MTV) and total lesion glycolysis (TLG) acquired from F-18 FDG PET/CT are predictors of prognosis of diffuse large B cell lymphoma (DLBCL) is controversial, with some studies concluding that it is and others concluding the opposite. Therefore, a systematic review was performed to explore the relationship of F-18 FDG PET/CT quantization parameters with the prognosis of DLBCL. Seven trials with a total of 703 DLBCL patients were included for analysis. Hazard ratios (HRs) for progression-free survival (PFS) and overall survival (OS), and odds ratios (ORs) for 3-year PFS and OS were pooled using the STATA package. Combined results suggested a strong link between the high SUVmax, MTV and TLG values and the poor 3-year PFS with ORs of 2.59, 3.69 and 2.29, respectively. Similarly, high MTV and TLG values unfavorably influenced the 3-year OS with ORs of 5.40 and 2.19, respectively. The pooled results also showed that high SUVmax and MTV were negative predictors of PFS with HRs of 1.61 (p = 0.038) and 2.18 (p = 0.000), respectively. The TLG value was not predictive of PFS. And for OS, only high MTV was a strong predictor of poor prognosis in DLBCL with HR 2.99 (p = 0.000). Our results suggested that SUVmax and MTV may be significant prognostic markers for PFS and MTV may be the only predictor for OS in DLBCL.


Atomistic dipole moments and polarizabilities of Na(N) clusters, N = 2-20.
The atomic-level response of Na(N) clusters, N = 2-20, to a small static external electric field is studied using a method that decomposes the total cluster dipole moment and polarizability into contributions from nonoverlapping atomic volumes. The atomic dipole moments and polarizabilities are, in turn, partitioned into the so-called dipole and charge-transfer components. The former characterizes a dielectric type of a response, whereas the latter represents a metallic type of a response. Analysis of the atomic polarizabilities points to their strong dependence on the site, or location, of the atoms within the structure of the clusters. Surface atoms have larger polarizabilities than the interior ones. Overall, the fraction of the charge-transfer component of the averaged atomic polarizabilities is an increasing function of the cluster size. The charge-transfer component is also responsible for the structure/shape driven variations in the atomic polarizabilities. The anisotropy of the total polarizabilities correlates with the shape anisotropy of the clusters.


Multistep Solid-State Organic Synthesis of Carbamate-Linked Covalent Organic Frameworks.
Herein, we demonstrate the first example of a multistep solid-state organic synthesis, in which a new imine-linked two-dimensional covalent organic framework (COF-170, 1) was transformed through three consecutive postsynthetic modifications into porous, crystalline cyclic carbamate and thiocarbamate-linked frameworks. These linkages are previously unreported and inaccessible through de novo synthesis. While not altering the overall connectivity of the framework, these chemical transformations induce significant conformational and structural changes at each step, highlighting the key importance of noncovalent interactions and conformational flexibility to COF crystallinity and porosity. These transformations were assessed using 15N multiCP-MAS NMR spectroscopy, providing the first quantitation of yields in COF postsynthetic modification reactions, as well as of amine defect sites in imine-linked COFs. This multistep COF linkage postsynthetic modification represents a significant step toward bringing the precision of organic solution-phase synthesis to extended solid-state compounds.


Role of catecholamines in release of gonadotrophic hormones in the rabbit.
Role of catecholamines (CA) in transmission of neurohormone releasing gonadotrophin (GnHR) from the hypothalamus to hypophysis was estimated on the basis of the hypothalamic catecholaminergic system blockade with 6-hydroxydopamine (6-OHDA) and intraventricular infusions of CA in the rabbit. 6-OHDA administered intraventricularly in doses 200-500 mug caused temporary blockade of ovulation. Intraventricular infusion of noradrenaline (NA) induced ovulation in 50% of animals tested, whereas dopamine (DA) and adrenaline (A), induced ovulation only in very few cases. It seems, that catecholaminergic system participates in transmission of neurohormones to the hypophysis, and NA plays the most important role in this process.


Effect of Zirconia Dental Implant Surfaces on Bone Integration: A Systematic Review and Meta-Analysis.
Background. The information available about osseointegration and the bone to implant interaction of zirconia implants with various surface modifications is still far from sufficient. Objective. The purpose of this systematic review and meta-analysis was to evaluate and compare zirconia dental implants with different surface topographies, with a focus on bone to implant contact and removal torque. Methods. The systematic review of the extracted publications was performed to compare the bone to implant contact (BIC) with removal torque (RT) values of titanium dental implants and machined and surfaced modified zirconia implants. Results. A total of fifteen articles on BIC and RT values were included in the quantitative analysis. No significant difference in the BIC values was observed between titanium and machined zirconia implants (p = 0.373; 95% CI: -0.166 to 0.443). However, a significantly better BIC values were observed for acid etched zirconia implants compared with those of titanium implants (p = 0.032; 95% CI: 0.068 to 1.461). Unmodified zirconia implants showed favorable BIC values compared to modified-surface zirconia implants (p = 0.021; 95% CI: -0.973 to -0.080). Conclusion. Acid etched zirconia implants may serve as a possible substitute for successful osseointegration.


Priorities in health care: reply to Lewis and Charny.
This paper is a reply to proposals to base priority health-care decisions on public opinion surveys. Whilst it is recognised that current practice is less than satisfactory, it is argued here that basing health-care priorities on societal attitudes in this way is not a solution and does not provide a satisfactory basis for bringing democracy to the health service.


Synthesis, antioxidant, and cytotoxic activities of N-azole substituted thiomorpholine derivatives.
A new class of N-azole substituted thiomorpholine derivatives were prepared and their antioxidant and cytotoxic activities were studied. The methyl substituted oxazolyl thiomorpholine dioxide 9b exhibited radical scavenging activity greater than the standard ascorbic acid. On the other hand, the thiazolyl thiomorpholine 10c having a chloro substituent on the aromatic ring was identified as a remarkable lead molecule for cytotoxic activity against A549 and HeLa cells, with IC50 values of 10.1 and 30.0 µM, respectively.


Multiple imputation for non-response when estimating HIV prevalence using survey data.
Missing data are a common feature in many areas of research especially those involving survey data in biological, health and social sciences research. Most of the analyses of the survey data are done taking a complete-case approach, that is taking a list-wise deletion of all cases with missing values assuming that missing values are missing completely at random (MCAR). Methods that are based on substituting the missing values with single values such as the last value carried forward, the mean and regression predictions (single imputations) are also used. These methods often result in potential bias in estimates, in loss of statistical information and in loss of distributional relationships between variables. In addition, the strong MCAR assumption is not tenable in most practical instances. Since missing data are a major problem in HIV research, the current research seeks to illustrate and highlight the strength of multiple imputation procedure, as a method of handling missing data, which comes from its ability to draw multiple values for the missing observations from plausible predictive distributions for them. This is particularly important in HIV research in sub-Saharan Africa where accurate collection of (complete) data is still a challenge. Furthermore the multiple imputation accounts for the uncertainty introduced by the very process of imputing values for the missing observations. In particular national and subgroup estimates of HIV prevalence in Zimbabwe were computed using multiply imputed data sets from the 2010-11 Zimbabwe Demographic and Health Surveys (2010-11 ZDHS) data. A survey logistic regression model for HIV prevalence and demographic and socio-economic variables was used as the substantive analysis model. The results for both the complete-case analysis and the multiple imputation analysis are presented and discussed. Across different subgroups of the population, the crude estimates of HIV prevalence are generally not identical but their variations are consistent between the two approaches (complete-case analysis and multiple imputation analysis). The estimates of standard errors under the multiple imputation are predominantly smaller, hence leading to narrower confidence intervals, than under the complete case analysis. Under the logistic regression adjusted odds ratios vary greatly between the two approaches. The model based confidence intervals for the adjusted odds ratios are wider under the multiple imputation which is indicative of the inclusion of a combined measure of the within and between imputation variability. There is considerable variation between estimates obtained between the two approaches. The use of multiple imputations allows the uncertainty brought about by the imputation process to be measured. This consequently yields more reliable estimates of the parameters of interest and reduce the chances of declaring significant effects unnecessarily (type I error). In addition, the utilization of the powerful and flexible statistical computing packages in R enhances the computations.


Early sepsis in Australia and New Zealand: A point-prevalence study of haemodynamic resuscitation practices.
Optimal resuscitation of sepsis-induced hypotension is uncertain, particularly the role of restrictive fluid strategies, leading to variability in usual practice. The objective of this study is to understand resuscitation practices in patients presenting to ED with early sepsis. Design, participants and setting: Prospective, observational, multicentre, single-day, point-prevalence study enrolling adult patients present in 51 Australian and New Zealand ICUs at 10.00 hours, 8 June 2021. Site-level data on sepsis policies and patient-level demographic data, presence of sepsis and fluid and vasopressor administration in the first 24 h post-ED presentation. A total of 722 patients were enrolled. ED was the ICU admission source for 222 of 722 patients (31.2%) and 78 of 222 patients (35%) met the criteria for sepsis within 24 h of ED presentation. Median age of the sepsis cohort was 61 (48-72) years, 58% were male and respiratory infection was the commonest cause (53.8%). The sepsis cohort had a higher severity of illness than the non-sepsis cohort (144/222 patients) and chronic immunocompromise was more common. Of 78 sepsis patients, 55 (71%) received ≥1 fluid boluses with 500 and 1000 mL boluses equally common (both 49%). In the first 24 h, 2335 (1409-3125) mL (25.3 [13.2-42.9] mL/kg) was administered. Vasopressors were administered in 53 of 78 patients (68%) and for 25 patients (47%) administration was peripheral. ICU patients presenting to the ED with sepsis receive less fluids than current international recommendations and peripheral vasopressor administration is common. This finding supports the conduct of clinical trials evaluating optimal fluid dose and vasopressor timing for early sepsis-induced hypotension.


Experiential Learning Methods for Biostatistics Students: A Model for Embedding Student Interns in Academic Health Centers.
This manuscript describes an experiential learning program for future collaborative biostatisticians (CBs) developed within an academic medical center. The program is a collaborative effort between the Biostatistics, Epidemiology, and Research Design (BERD) Methods Core and the Master of Biostatistics (MB) program, both housed in the Department of Biostatistics and Bioinformatics at Duke University School of Medicine and supported in partnership with the Duke Clinical and Translational Science Institute. To date, the BERD Core Training and Internship Program (BCTIP) has formally trained over 80 students to work on collaborative teams that are integrated throughout the Duke School of Medicine. This manuscript focuses on the setting for the training program, the experiential learning model on which it is based, the structure of the program, and lessons learned to date.


The endemic treponematoses.
The agents of human treponematoses include four closely related members of the genus Treponema: three subspecies of Treponema pallidum plus Treponema carateum. T. pallidum subsp. pallidum causes venereal syphilis, while T. pallidum subsp. pertenue, T. pallidum subsp. endemicum, and T. carateum are the agents of the endemic treponematoses yaws, bejel (or endemic syphilis), and pinta, respectively. All human treponematoses share remarkable similarities in pathogenesis and clinical manifestations, consistent with the high genetic and antigenic relatedness of their etiological agents. Distinctive features have been identified in terms of age of acquisition, most common mode of transmission, and capacity for invasion of the central nervous system and fetus, although the accuracy of these purported differences is debated among investigators and no biological basis for these differences has been identified to date. In 2012, the World Health Organization (WHO) officially set a goal for yaws eradication by 2020. This challenging but potentially feasible endeavor is favored by the adoption of oral azithromycin for mass treatment and the currently focused distribution of yaws and endemic treponematoses and has revived global interest in these fascinating diseases and their causative agents.


[The influence of pain aliments and vertebral mobility in patients after whole body cryotherapy].
The motive efficiency depends from many factors, in this currently also form the state of health. The change in grounds of vertebral column causing the limitations of mobility and pain can significance worsen the functional state, motive efficiency and quality of patient's life. The leadership of active mode of life complying to recommendations the preventive style of life as well as the using with interventions of physical medicine influences on improvement significance the state of health. It the examination groups were 42 patients (21 women and 21 men) in age from 45-65 years with low back pain about different etiology. All studied were thirty interventions of whole body cryotherapy and then practices kinesitherapy exercises by 30 minutes subjected on gymnastic room. To evaluation of stepping pain ailments was the scale VAS executed, mobility of lumbar spine with help of Schober test as well as was executed. All results were statistical analysis subjected. After whole body cryotherapy the pain ailments in scale VAS were statistical significant in men group. The decrease of pain ailments influenced on improvement column mobility vertebral spine with use a Schober test (p<0,05). The statistical analysis were not significant in women group of pain ailments and the opinion of mobility of vertebral column spine (p > 0,05). Whole body cryotherapy reduced significance pain ailments and mobility of lumbar spine in men group. In women group the got results were not statistical significant.


The effects of feeding suet-enriched chow on site-specific differences in the composition of triacylglycerol fatty acids in adipose tissue and its interactions in vitro with lymphoid cells.
The effects of diet on the composition and properties of adipose tissue in relation to lymph nodes were studied in adult guinea-pigs. The proportions of monoenoic triacylglycerol fatty acids were constant in all sites in adipose tissue of similarly fed guinea-pigs, but were substantially greater in samples from guinea-pigs fed on suet-enriched chow. Triacylglycerols in adipose tissue from near nodes contained significantly fewer saturated fatty acids, and significantly more 18:2n-6 and 18:3n-3 than those in samples from sites remote from nodes within the same depot. Depots that interact most strongly with lymphoid cells in vitro had the largest and most consistent within-depot differences. The gradients of triacylglycerol fatty acid composition with distance from lymph nodes in two small intermuscular depots were similar in guinea-pigs fed on plain or suet-enriched chow. These findings are consistent with the hypothesis that adipose tissue around lymph nodes is specialized for local interactions with the lymphoid cells therein, and help to explain the variability of serial or duplicate measurements of adipose tissue composition. When cultured alone, lipopolysaccharide-stimulated lymph node lymphoid cells from suet-fed guinea-pigs incorporated as much labelled thymidine as the controls. Adipose tissue explants from suet-fed guinea-pigs inhibited lymphocyte proliferation much less than those of the controls, although the site-specific differences were similar. The pattern of site-specific differences in glycerol released from explants incubated alone was generally similar for both dietary groups, but except in the popliteal depot, the increases following co-culturing with lymphoid cells were smaller for samples from suet-fed guinea-pigs. These experiments show that minor changes in the fatty acid composition of the diet can substantially alter the interactions between adipose tissue and lymphoid cells.


Novel Ti-O-Ti bonding species constructed in a metal-oxide cluster: reaction products of bis(oxalato)oxotitanate(IV) with the dimeric, 1,2-dititanium(IV)-substituted Keggin polyoxotungstate.
The preparation and structural characterization of a novel Ti-O-Ti bonding complex constructed in a dilacunary alpha-Keggin polyoxometalate (POM), [[{Ti(ox)(H2O)}4(mu-O)3](alpha-PW10O37)](7-) (H2ox = oxalic acid) (1a), are described. The water-soluble, crystalline complex with a formula of K6H[1a].0.5KCl.10H2O (1p) was prepared as the bulk sample in 28.0% (0.51 g scale) yield in a 1:4 molar-ratio reaction of the dititanium(IV)-substituted, dimeric form of an alpha-Keggin POM, K10[(alpha-1,2-PW10Ti2O39)2].18H2O, with the titanium(IV) source K2[TiO(ox)2].2H2O in HCl-acidic solution (pH 0.08). Prior to formation of 1p, the KCl-free crystalline compound (1c) obtained was characterized with X-ray crystallography. The compound 1p was unequivocally characterized with complete elemental analysis, thermogravimetric and differential thermal analyses (TG/DTA), FTIR, and solution (31P, 183W, and 13C) NMR spectroscopy. The molecular structure of 1a was determined. The POM 1a in the solid state was composed of the four octahedral Ti groups (four guests), i.e., the two Ti-O-Ti groups linked with the mu-O atom, incorporated to the two adjacent, octahedral vacant sites (two hosts) in the dilacunary Keggin POM. The formation of 1a, as well as the recently found POM [{Ti(ox)(H2O)}2(mu-O)](alpha-PW11O39)](5-) (2a), was strongly dependent on the reaction with [TiO(ox)2](2-), i.e., the anionic titanium(IV) complex as the titanium(IV) source. The POM 1a is contrasted to most titanium(IV)-substituted POMs consisting of a combination of a monolacunary site (one host) and an octahedral Ti group (one guest) and also contrasted to 2a as a combination of a monolacunary site (one host) and two octahedral Ti groups or a Ti-O-Ti group (two guests).


Critical period for cross-modal plasticity in blind humans: a functional MRI study.
The primary visual cortex (V1) in congenitally blind humans has been shown to be involved in tactile discrimination tasks, indicating that there is a shift in function of this area of cortex, but the age dependency of the reorganization is not fully known. To investigate the reorganized network, we measured the change of regional cerebral blood flow using 3.0 Tesla functional MRI during passive tactile tasks performed by 15 blind and 8 sighted subjects. There was increased activity in the postcentral gyrus to posterior parietal cortex and decreased activity in the secondary somatosensory area in blind compared with sighted subjects during a tactile discrimination task. This suggests that there is a greater demand for shape discrimination processing in blind subjects. Blind subjects, irrespective of the age at onset of blindness, exhibited higher activity in the visual association cortex than did sighted subjects. V1 was activated in blind subjects who lost their sight before 16 years of age, whereas it was suppressed in blind subjects who lost their sight after 16 years of age during a tactile discrimination task. This suggests that the first 16 years of life represent a critical period for a functional shift of V1 from processing visual stimuli to processing tactile stimuli. Because of the age-dependency, V1 is unlikely to be the "entry node" of the cortex for the redirection of tactile signals into visual cortices after blinding. Instead, the visual association cortex may mediate the circuitry by which V1 is activated during tactile stimulation.


Analytical Techniques and Pharmacokinetics of Gastrodia elata Blume and Its Constituents.
Gastrodia elata Blume (G. elata), commonly called Tianma in Chinese, is an important and notable traditional Chinese medicine (TCM), which has been used in China as an anticonvulsant, analgesic, sedative, anti-asthma, anti-immune drug since ancient times. The aim of this review is to provide an overview of the abundant efforts of scientists in developing analytical techniques and performing pharmacokinetic studies of G. elata and its constituents, including sample pretreatment methods, analytical techniques, absorption, distribution, metabolism, excretion (ADME) and influence factors to its pharmacokinetics. Based on the reported pharmacokinetic property data of G. elata and its constituents, it is hoped that more studies will focus on the development of rapid and sensitive analytical techniques, discovering new therapeutic uses and understanding the specific in vivo mechanisms of action of G. elata and its constituents from the pharmacokinetic viewpoint in the near future. The present review discusses analytical techniques and pharmacokinetics of G. elata and its constituents reported from 1985 onwards.


Serine protease EspP from enterohemorrhagic Escherichia coli is sufficient to induce shiga toxin macropinocytosis in intestinal epithelium.
Life-threatening intestinal and systemic effects of the Shiga toxins produced by enterohemorrhagic Escherichia coli (EHEC) require toxin uptake and transcytosis across intestinal epithelial cells. We have recently demonstrated that EHEC infection of intestinal epithelial cells stimulates toxin macropinocytosis, an actin-dependent endocytic pathway. Host actin rearrangement necessary for EHEC attachment to enterocytes is mediated by the type 3 secretion system which functions as a molecular syringe to translocate bacterial effector proteins directly into host cells. Actin-dependent EHEC attachment also requires the outer membrane protein intimin, a major EHEC adhesin. Here, we investigate the role of type 3 secretion in actin turnover occurring during toxin macropinocytosis. Toxin macropinocytosis is independent of EHEC type 3 secretion and intimin attachment. EHEC soluble factors are sufficient to stimulate macropinocytosis and deliver toxin into enterocytes in vitro and in vivo; intact bacteria are not required. Intimin-negative enteroaggregative Escherichia coli (EAEC) O104:H4 robustly stimulate Shiga toxin macropinocytosis into intestinal epithelial cells. The apical macropinosomes formed in intestinal epithelial cells move through the cells and release their cargo at these cells' basolateral sides. Further analysis of EHEC secreted proteins shows that a serine protease EspP alone is able to stimulate host actin remodeling and toxin macropinocytosis. The observation that soluble factors, possibly serine proteases including EspP, from each of two genetically distinct toxin-producing strains, can stimulate Shiga toxin macropinocytosis and transcellular transcytosis alters current ideas concerning mechanisms whereby Shiga toxin interacts with human enterocytes. Mechanisms important for this macropinocytic pathway could suggest new potential therapeutic targets for Shiga toxin-induced disease.


Genetic Characterization of the Soybean Nested Association Mapping Population.
A set of nested association mapping (NAM) families was developed by crossing 40 diverse soybean [ (L.) Merr.] genotypes to the common cultivar. The 41 parents were deeply sequenced for SNP discovery. Based on the polymorphism of the single-nucleotide polymorphisms (SNPs) and other selection criteria, a set of SNPs was selected to be included in the SoyNAM6K BeadChip for genotyping the parents and 5600 RILs from the 40 families. Analysis of the SNP profiles of the RILs showed a low average recombination rate. We constructed genetic linkage maps for each family and a composite linkage map based on recombinant inbred lines (RILs) across the families and identified and annotated 525,772 high confidence SNPs that were used to impute the SNP alleles in the RILs. The segregation distortion in most families significantly favored the alleles from the female parent, and there was no significant difference of residual heterozygosity in the euchromatic vs. heterochromatic regions. The genotypic datasets for the RILs and parents are publicly available and are anticipated to be useful to map quantitative trait loci (QTL) controlling important traits in soybean.


[The pathogens and their antibiotic sensitivities in hospital-acquired pneumonia cases].
Dynamic precautions in the early diagnosis and treatment of hospital acquired pneumonias are necessary because of their high mortality. In these patients, invasive diagnostic approaches may be needed since clinical and radiological findings and other non-invasive approaches frequently fail to establish the diagnosis. Thirty eight patients were prospectively included in the study. Average age of patients was 45.5 +/- 16.4 years; 31 were males (81.6%) and 7 (18.4%) were females. Pneumonia was detected in 9 (23.7%) cases during the first five days and in 29 (76.3%) cases after the fifth day of admission to the hospital. Bronchoscopic interventions diagnostic purpose were carried out in 25 (65.8%) patients. The culture results were negative in 7 (18.4%) cases. While more than one pathogen was determined on the cultures of 16 (42.1%) patients only one pathogen was isolated in the cultures of 15 (39.4%) cases. The frequently isolated pathogen on cultures was Staphylococci (45.4%). Other pathogens were Enterobacter spp., Pseudomonas spp., Escherichia coli, Serratia and Streptococcus pneumoniae according to their frequency on cultures. High resistance rates to the third generation cephalosporins were determined. Eleven of 17 deaths in 38 pneumonia cases were attributable to pneumonia. As a conclusion, isolation of pathogen and antibiotic resistance should be determined in the cases with hospital acquired pneumonia. Invasive diagnostic interventions were not avoided when necessary. Although pro-BAL and PSB methods were expensive, their use in selected cases may prevent unnecessary antibiotic use and contribute to a decrease in mortality rate.


Recurrent stupor due to lysinuric protein intolerance.
Recurrent stupor in children is an uncommon clinical problem with a wide differential diagnosis; inherited metabolic disorders account for a vast majority. We report a 9-year-old girl with recurrent episodes of stupor. Initial episode was treated as viral encephalitis and the second episode was managed as non-convulsive status epilepticus. Hyperammonemia was detected in the last episode. Metabolic work-up after dietary protein challenge revealed classical biochemical features of lysinuric protein intolerance. She was managed with protein-restricted diet, which resulted in marked neurological improvement. LPI is a rare inherited metabolic disorder due to membrane transport defect of cationic amino acids.


Mutations of the thyroid peroxidase gene in Chinese siblings with congenital goitrous hypothyroidism.
To investigate thyroid peroxidase gene (TPO) mutations in a Chinese siblings with congenital goitrous hypothyroidism (CGH). The proband, his sister, and their parents were enrolled. All subjects underwent clinical examination and laboratory tests. Mutation screening of the TPO gene was performed by sequencing fragments amplified from extracted genomic DNA. The siblings were diagnosed as CGH with neurodevelopmental deficits. Two compound heterozygous inactivating mutations were found in the two patients: a frameshift mutation between positions 2268 and 2269 (c.2268-2269 insT) and a missense mutation at c.2089 G>A (p.G667S) of the TPO gene. Their parents, with normal thyroid hormone levels, were heterozygous for mutations c.2268-2269 insT and c.2089 G>A, respectively. The polymorphisms of c.1207 G>T, c.1283 G>C, and c.2088 C>T were detected in the family. CGH of the Chinese siblings was due to the TPO gene mutations (c.2268-2269 insT and c.2089 G>A).


Colchicine in the treatment of scleroderma.
We treated 10 patients with well established scleroderma with colchicine in the highest tolerated dose for one year. We could determine no clinical or laboratory improvement in any patient. Progression of disease was suggested by the development of new skin ulcers and telangiectasia, by an increase in musculoskeletal complaints, and by a significantly diminished ability to make a fist. In addition, there was deterioration of pulmonary function as demonstrated by a statistically significant fall in FEV1, and diffusing capacity. Colchicine was not of value in the treatment of scleroderma in this group of patients.


Permanent gentamicin vestibulotoxicity.
To determine the natural history of permanent gentamicin vestibulotoxicity. Retrospective; comparison of retrospective and prospective studies. Tertiary neurotology clinic. Clinical research and technology center. Thirty-three subjects with permanent gentamicin-induced vestibulotoxicity. Medical records review, neurotologic examination, and vestibular and auditory function tests. Vestibular and auditory function test results at least 1 year after discontinuation of gentamicin, clinical examination results, serum gentamicin levels, and serum creatinine levels. Thirty-three subjects had vestibular function test results consistent with permanent gentamicin ototoxicity. All complained of dysequilibrium, 32 described oscillopsia, and 23 had tinnitus. All 33 subjects had complained of symptoms consistent with ototoxicity within 1 to 3 weeks of initiation of gentamicin therapy; however, gentamicin vestibulotoxicity was not recognized before hospital discharge in 32 of 33 subjects. Serum peak and trough gentamicin levels did not correlate with the development of vestibulotoxicity, nor did observance of recommended "safe" dosage ranges. Of 17 subjects whose serum creatinine levels were recorded, 6 experienced abnormal elevations in serum creatinine in conjunction with gentamicin use. Gentamicin can cause permanent vestibular and auditory ototoxicity. There is no safe dose of gentamicin. Serum gentamicin levels are of no value in predicting the onset, occurrence, or severity of vestibulotoxicity or cochleotoxicity. Termination of gentamicin on appearance of signs or symptoms of ototoxicity may reduce the incidence of permanent vestibular ototoxicity. When possible, other antibiotics should be administered.


Phosphorodiamidate-directed metalation of N-heterocycles using Mg- and Zn-TMP bases.
The strong directing ability of the N,N,N',N'-tetramethyldiaminophosphorodiamidate group has been used to achieve selective metalations on various heterocycles such as pyridines, quinolines and quinoxalines with TMP-derived bases like TMPMgCl x LiCl, TMP(2)Mg x 2 LiCl, and TMP(2)Zn x 2 MgCl(2) x 2 LiCl. This protocol was applied in the synthesis of etoricoxib, talnetant and a P-selectin inhibitor.


Structure-affinity properties of a high-affinity ligand of FKBP12 studied by molecular simulations of a binding intermediate.
With a view to explaining the structure-affinity properties of the ligands of the protein FKBP12, we characterized a binding intermediate state between this protein and a high-affinity ligand. Indeed, the nature and extent of the intermolecular contacts developed in such a species may play a role on its stability and, hence, on the overall association rate. To find the binding intermediate, a molecular simulation protocol was used to unbind the ligand by gradually decreasing the biasing forces introduced. The intermediate was subsequently refined with 17 independent stochastic boundary molecular dynamics simulations that provide a consistent picture of the intermediate state. In this state, the core region of the ligand remains stable, notably because of the two anchoring oxygen atoms that correspond to recurrent motifs found in all FKBP12 ligand core structures. Besides, the non-core regions participate in numerous transient intermolecular and intramolecular contacts. The dynamic aspect of most of the contacts seems important both for the ligand to retain at least a part of its configurational entropy and for avoiding a trapped state along the binding pathway. Since the transient and anchoring contacts contribute to increasing the stability of the intermediate, as a corollary, the dissociation rate constant [Formula: see text] of this intermediate should be decreased, resulting in an increase of the affinity constant [Formula: see text]. The present results support our previous conclusions and provide a coherent rationale for explaining the prevalence in high-affinity ligands of (i) the two oxygen atoms found in carbonyl or sulfonyl groups of dissimilar core structures and of (ii) symmetric or pseudo-symmetric mobile groups of atoms found as non-core moieties. Another interesting aspect of the intermediate is the distortion of the flexible 80 s loop of the protein, mainly in its tip region, that promotes the accessibility to the bound state.