Increased instability of intermediate alleles in families with sporadic Huntington disease compared to similar sized intermediate alleles in the general population.
We have directly compared intergenerational stability of intermediate alleles (IAs) derived from new mutation families (IANM) for Huntington disease (HD) with IAs in the general population (IAGP) which occur in approximately 1 in 50 persons. Analysis of meiotic events in blood and sperm reveals that IANM are significantly more unstable than IAGP despite similar size. However, for both IANM and IAGP CAG changes were small and risks for inheriting an expansion into the HD affected range were low. Sequence analysis reveals that the CAG tract is generally interrupted by a penultimate CAA in IAGP, IANM and alleles in the affected range. In one new mutation family, however, two A-->G mutations result in a pure CAG tract which is associated with very marked instability. These mutations alter the predicted DNA hairpin structure with a predicted increase in the likelihood of large expansion, supporting the model that hairpin loop formation plays an important role in trinucleotide instability.


Unravelling the mechanism of amyloid-β peptide oligomerization and fibrillation at chiral interfaces.
In this communication, the mechanism of how surface chirality affects amyloid-β (Aβ) fibrillation was firstly unravelled at the molecular level: a chiral surface serves to control the 2D-diffusion and surface residence time of Aβ molecules via the chiral recognition with Aβ to allow precursor Aβ to laterally diffuse and collide with each other for oligomerization and fibrillation. Surface chirality that shortens the surface residence time of Aβ, for example, R-cysteine modification with carboxylic, secondary amine and thiol groups surrounding the chiral center, can retard Aβ oligomerization and fibrillation. This work is essential to a deeper fundamental understanding of the effects of surface chirality on amyloidosis processes as well as the development of chiral materials to inhibit Aβ fibrillation.


Exponential-phase expression of spvA of the Salmonella typhimurium virulence plasmid: induction in intracellular salts medium and intracellularly in mice and cultured mammalian cells.
The spv genes of Salmonella typhimurium and other non-typhoidal Salmonella serovars are essential for efficient systemic infection beyond the intestines in orally inoculated mice as a model for enteric fever. These virulence genes are not significantly expressed by salmonellae during exponential growth in L broth but are induced when the bacteria enter the stationary phase of growth. Using RNase protection analysis to directly measure spvA mRNA from the virulence plasmid of S. typhimurium, we found that spvA was maximally induced in an SpvR- and RpoS-dependent manner during exponential growth in intracellular Salts Medium, which mimics the intracellular environment of mammalian cells. A cloned spvA-lacZ operon fusion in S. typhimurium was induced intracellularly in periotoneal cells of mice, correlating in vivo intracellular gene expression with intracellular function of the spv genes in infected mice. spvA was also induced intracellularly in vitro within both Henle-407 intestinal epithelial cells and J774.A1 macrophage-like cells when the bacteria were replicating with exponential kinetics. Prevention of invasion of salmonellae with cytochalasin D inhibited spvA induction within tissue culture cells, indicating that salmonellae must be internalized for spvA to be induced. The spvA-lacZ fusion was not induced by salmonellae in extracellular fluid of the peritoneal cavity or in serum. Since induction of the spv genes occurs intracellularly during exponential growth of salmonellae, cessation of growth may not be the most relevant inducing signal for spv gene expression.


Decreased susceptibility to calpains of v-FosFBR but not of v-FosFBJ or v-JunASV17 retroviral proteins compared with their cellular counterparts.
The c-Fos and c-Jun transcription factors are rapidly turned over in vivo. One of the multiple pathways responsible for their breakdown is probably initiated by calpains, which are cytoplasmic calcium-dependent cysteine proteases. The c-fos gene has been transduced by two murine oncogenic retroviruses called Finkel-Biskis-Jenkins murine sarcoma virus (FBJ-MSV) and Finkel-Biskis-Reilly murine sarcoma virus (FBR-MSV); c-jun has been transduced by the chicken avian sarcoma virus 17 (ASV17) retrovirus. Using an in vitro degradation assay, we show that the mutated v-FosFBR, but not v-FosFBJ or v-JunASV17, is resistant to calpains. This property raises the interesting possibility that decreased sensitivity to calpains might contribute to the tumorigenic potential of FBR-MSV by allowing greater accumulation of the protein that it encodes in infected cells. It has also been demonstrated that resistance to cleavage by calpains does not result from mutations that have accumulated in the Fos moiety of the viral protein but rather from the addition of atypical peptide motifs at its both ends. This observation raises the interesting possibility that homologous regions in viral and cellular Fos either display slightly different conformations or are differentially accessible to interacting proteins.


Transport and retention of engineered silver nanoparticles in carbonate-rich sediments in the presence and absence of soil organic matter.
The transport and retention behavior of polymer- (PVP-AgNP) and surfactant-stabilized (AgPURE) silver nanoparticles in carbonate-dominated saturated and unconsolidated porous media was studied at the laboratory scale. Initial column experiments were conducted to investigate the influence of chemical heterogeneity (CH) and nano-scale surface roughness (NR) arising from mixtures of clean, positively charged calcium carbonate sand (CCS), and negatively charged quartz sands. Additional column experiments were performed to elucidate the impact of CH and NR arising from the presence and absence of soil organic matter (SOM) on a natural carbonate-dominated aquifer material. The role of the nanoparticle capping agent was examined under all conditions tested in the column experiments. Nanoparticle transport was well described using a numerical model that facilitated blocking on one or two retention sites. Results demonstrate that an increase in CCS content in the artificially mixed porous medium leads to delayed breakthrough of the AgNPs, although AgPURE was much less affected by the CCS content than PVP-AgNPs. Interestingly, only a small portion of the solid surface area contributed to AgNP retention, even on positively charged CCS, due to the presence of NR which weakened the adhesive interaction. The presence of SOM enhanced the retention of AgPURE on the natural carbonate-dominated aquifer material, which can be a result of hydrophobic or hydrophilic interactions or due to cation bridging. Surprisingly, SOM had no significant impact on PVP-AgNP retention, which suggests that a reduction in electrostatic repulsion due to the presence of SOM outweighs the relative importance of other binding mechanisms. Our findings are important for future studies related to AgNP transport in shallow unconsolidated calcareous and siliceous sands.


Incidence of Duchenne muscular dystrophy in New South Wales and Australian Capital Territory.
A survey was conducted during 1977 to 1978 to determine the incidence of Duchenne muscular dystrophy in New South Wales and the Australian Capital Territory. Ninety-nine cases born between 1960 and 1971 were ascertained representing an incidence rate of 18.6 per 100 000 liveborn males. Over 60% of the ascertained males were isolated cases, and only 7% of affected males had an affected males relative in a previous generation in the female line. Categorisation of ascertained cases as either "theoretically preventable" or not shows that genetic counselling alone in the present circumstances is insufficient to reduce the incidence rate significantly.


ATP signalling in epilepsy.
This paper focuses on a role for ATP neurotransmission and gliotransmission in the pathophysiology of epileptic seizures. ATP along with gap junctions propagates the glial calcium wave, which is an extraneuronal signalling pathway in the central nervous system. Recently astrocyte intercellular calcium waves have been shown to underlie seizures, and conventional antiepileptic drugs have been shown to attenuate these calcium waves. Blocking ATP-mediated gliotransmission, therefore, represents a potential target for antiepileptic drugs. Furthermore, while knowledge of an antiepileptic role for adenosine is not new, a recent study showed that adenosine accumulates from the hydrolysis of accumulated ATP released by astrocytes and is believed to inhibit distant synapses by acting on adenosine receptors. Such a mechanism is consistent with a surround-inhibitory mechanism whose failure would predispose to seizures. Other potential roles for ATP signalling in the initiation and spread of epileptiform discharges may involve synaptic plasticity and coordination of synaptic networks. We conclude by making speculations about future developments.


The effect of shoe toe box shape and volume on forefoot interdigital and plantar pressures in healthy females.
Ill-fitting footwear can be detrimental to foot health with the forefoot being an area for most discomfort. Studies on footwear have primarily examined sports or orthopaedic prescription shoes and little is known about the effects that everyday flat shoes have on the forefoot. The aim of this study was to investigate the effect of toe box shape in a popular slip-on pump on dorsal and plantar pressures with particular interest around the forefoot in a healthy female population. A convenience sample of 27 female participants with no known foot pathologies was recruited. After assessment of foot size, plantar foot pressure and interdigital pressures were recorded for each of the 3 different toe box styles; round, square and pointed. Participants walked at a self-selected speed over a 10 m walkway whilst wearing each of the 3 styles of shoe and also whilst barefoot. Processed and analysed data extracted included peak pressure, time to peak pressure, contact time and pressure time integral. ANOVA and Freidman analysis was used to test for statistical significance. Shoes with a round toe showed least pressure around the medial aspect of the toes whilst the pointed shoe had least pressure on the lateral toes. Contact times for the plantar regions were not altered in any shoe condition yet contact around the medial aspect of the toes was highest in the pointed shoe. This study highlights that the shape of the toe box in footwear can significantly influence the amount of pressure applied to the forefoot. Furthermore, the contours of the shoe also have an impact on the contact time and pressure time integral around the forefoot and also the peak plantar pressure in the toe region. The changes observed could be significant in the development of pathology in certain footwear toe box shapes. Consideration should be given to footwear design around the toe box to improve fit and reduce pressure. Further work is required to investigate the effect of toe box shape and volume on a pathological population with pressure related lesions.


A Direct in Vivo Comparison of the Melanocortin Monovalent Agonist Ac-His-DPhe-Arg-Trp-NH2 versus the Bivalent Agonist Ac-His-DPhe-Arg-Trp-PEDG20-His-DPhe-Arg-Trp-NH2: A Bivalent Advantage.
Bivalent ligands targeting putative melanocortin receptor dimers have been developed and characterized in vitro; however, studies of their functional in vivo effects have been limited. The current report compares the effects of homobivalent ligand CJL-1-87, Ac-His-DPhe-Arg-Trp-PEDG20-His-DPhe-Arg-Trp-NH2, to monovalent ligand CJL-1-14, Ac-His-DPhe-Arg-Trp-NH2, on energy homeostasis in mice after central intracerebroventricular (ICV) administration into the lateral ventricle of the brain. Bivalent ligand CJL-1-87 had noteworthy advantages as an antiobesity probe over CJL-1-14 in a fasting-refeeding in vivo paradigm. Treatment with CJL-1-87 significantly decreased food intake compared to CJL-1-14 or saline (50% less intake 2-8 h after treatment). Furthermore, CJL-1-87 treatment decreased the respiratory exchange ratio (RER) without changing the energy expenditure indicating that fats were being burned as the primary fuel source. Additionally, CJL-1-87 treatment significantly lowered body fat mass percentage 6 h after administration (p < 0.05) without changing the lean mass percentage. The bivalent ligand significantly decreased insulin, C-peptide, leptin, GIP, and resistin plasma levels compared to levels after CJL-1-14 or saline treatments. Alternatively, ghrelin plasma levels were significantly increased. Serum stability of CJL-1-87 and CJL-1-14 (T1/2 = 6.0 and 16.8 h, respectively) was sufficient to permit physiological effects. The differences in binding affinity of CJL-1-14 compared to CJL-1-87 are speculated as a possible mechanism for the bivalent ligand's unique effects. We also provide in vitro evidence for the formation of a MC3R-MC4R heterodimer complex, for the first time to our knowledge, that may be an unexploited neuronal molecular target. Regardless of the exact mechanism, the advantageous ability of CJL-1-87 compared to CJL-1-14 to increase in vitro binding affinity, increase the duration of action in spite of decreased serum stability, decrease in vivo food intake, decrease mice's body fat percent, and differentially affect mouse hormone levels demonstrates the distinct characteristics achieved from the current melanocortin agonist bivalent design strategy.


Use of environmental scanning electron microscopy to evaluate dental stain removal.
The purpose of the study was to assess the usefulness of environmental scanning electron microscopy (ESEM) to evaluate stain removal from extracted teeth. The ESEM differs from conventional SEM in that no sample preparation is needed, eliminating artifactual changes. Furthermore, the same sample can be viewed on multiple occasions, allowing "before" and "after" pictures of the same tooth. As a model stain removal device, we tested the Sonicare sonic toothbrush, which has previously been shown to remove dental stain in vivo. Twelve freshly extracted teeth with extrinsic coffee, tea or tobacco stain were obtained for the study. Nine of these had heavy stain (stain covering more than one-third buccal or lingual surface) and were used without further modification. Three teeth were treated in vitro with chlorhexidine and a mixture of coffee and tea to enhance staining. All teeth were examined by ESEM at three times: prior to brushing, after 15-30 seconds of brushing, and after 60-80 seconds of brushing. Light microscopy and 35 mm photography was also done to correlate the ultrastructural changes with those visible at low magnification. Water, mouthwash and 30% slurry of toothpaste were used as fluid vehicles during brushing, but little difference in stain removal was noted among these three fluids. Approximately half the stain was removed within 15-30 seconds, and most visible stain was removed in 60-80 seconds of brushing. Pits and crevices of tooth enamel that were smaller than the bristle diameter, and thus would be inaccessible to abrasive cleaning by direct bristle contact, were generally found to be stain-free. These findings confirm previous reports of the stain removal effectiveness of the Sonicare, and demonstrate the usefulness of ESEM for stain removal studies.


Effects of dietary true digestible calcium to phosphorus ratio on growth performance and efficiency of calcium and phosphorus use in growing pigs fed corn and soybean meal-based diets.
Objectives of this study were to determine effects of dietary true fecal digestible Ca to true digestible P ratio on growth performance and efficiency of Ca and P use in growing pigs fed corn (Triticum aestivum)-soybean (Glycine max) meal (SBM)-based diets. Experiment 1 was carried out to measure true fecal digestibility of Ca and P as well as the fecal endogenous outputs of these nutrients associated with a corn and SBM-based diet in 12 Yorkshire growing pigs with an average initial BW of 23.2 ± 0.6 kg by the substitution method. True fecal digestibility values (%; n = 6) of Ca (53.6 ± 12.7) and P (43.8 ± 16.7) as well as the fecal endogenous outputs (g/kg DMI; n = 12) of Ca (0.91 ± 0.20) and P (1.31 ± 0.15) associated with the diets were determined. Experiment 2 was conducted with 36 Yorkshire barrows of an average initial BW of 24.2 ± 0.6 kg and the pigs were fed 6 diets according to a completely randomized block design. The 6 diets were corn and SBM based with diet 1 containing 0.2% true digestible Ca and 0.3% true digestible P and were formulated to contain 6 total Ca to total P ratios based on analyzed dietary Ca and P contents (diet 1, 0.6:1; diet 2, 0.7:1; diet 3, 0.8:1; diet 4, 1.3:1; diet 5, 1.0:1; and diet 6, 1.3:1) by supplementing gradient levels of limestone with a constant dietary P content for meeting the recommended requirement. Changes in the dietary Ca to P ratio had no effects (P > 0.05) on ADG. No differences (P > 0.05) in ADFI were observed between the other diets except the lower ADFI (P < 0.05) in diet 3 compared with diet 2. However, G:F was higher (P < 0.05) in diet 2 compared with diets 5 and 6. Changes in the dietary Ca to P ratio had consistent effects on true fecal P digestibility and retention with much lower values (P < 0.05) observed in diet 5 in comparison with the other diets. In summary, true fecal digestible Ca to P ratios of 0.9:1 to 1.0:1 were associated with optimal responses in both G:F as well as true fecal P digestibility and retention in growing pigs fed corn and SBM-based diets.


Direct immunofluorescence in recurrent aphthous ulcers and Behçet's disease.
Direct immunofluorescence (IF) investigations in oral aphthae were performed in 17 patients with recurrent aphthous ulcers (RAU) and in 9 patients with Behçet's disease (BD). In addition, cutaneous hyperreactivity lesions from 2 BD patients were examined. The results were compared to direct IF findings in 28 patients with nonaphthous oral diseases. There was no difference between direct IF on oral aphthae in RAU compared to BD. Deposition of C3 in vessel walls of the subepithelial connective tissue was observed in 13 of 17 patients with RAU and in all patients with BD. IgM deposits in vessel walls were found in 5 RAU and 3 BD patients. Deposition of C1q or C4 was not present. By contrast, IgG and/or IgM, C1q, C3 and C4 were detected in dermal vessel walls of skin hyperreactivity lesions from 2 BD patients. This finding is considered to be a true immune complex vasculitis. Compared to nonaphthous oral lesions, oral aphthae of BD and RAU were characterized by C3 deposition in the subepithelial vessel walls.


Synthesis, characterization and biological activity of new cyclometallated platinum(iv) iodido complexes.
The synthesis of six novel cyclometallated platinum(iv) iodido complexes is accomplished by intermolecular oxidative addition of methyl iodide (compounds 2a-2c) or iodine (compounds 3a-3c) upon cyclometallated platinum(ii) compounds [PtX{(CH3)2N(CH2)3NCH(4-ClC6H3)}] (1a-1c: X = Cl, CH3 or I). The X-ray molecular structures of platinum(ii) compound 1c and platinum(iv) compounds 3b and 3a' (an isomer of 3a) are reported. The cytotoxic activity against a panel of human adenocarcinoma cell lines (A-549 lung, MDA-MB-231 and MCF-7 breast, and HCT-116 colon), DNA interaction, topoisomerase I, IIα, and cathepsin B inhibition, and cell cycle arrest, apoptosis and ROS generation of the investigated complexes are presented. Remarkable antiproliferative activity was observed for most of the synthesized cycloplatinated compounds (series 1-3) in all the selected carcinoma cell lines. The best inhibition was provided for the octahedral platinum(iv) compounds 2a-2c exhibiting a methyl and an iodido axial ligand. Preliminary biological results point to a different mechanism of action for the investigated compounds. Cyclometallated platinum(ii) compounds 1a-1c modify the DNA migration as cisplatin. In contrast, cyclometallated platinum(iv) compounds 2a-2c and 3a-3c did not modify the DNA tertiary structure neither in the absence nor in the presence of ascorbic acid, which made them incapable of reducing platinum(iv) compounds 2b and 2c in a buffered aqueous medium (pH 7.40) according to 1H NMR experiments. Remarkable topoisomerase IIα inhibitory activity is reported for platinum(iv) complexes 2b and 3a and in addition, for the last one, a moderate cathepsin B inhibition is reported. Cell cycle arrest (decrease in G0/G1 and G2 phases and arrest in the S phase), induction of apoptosis and ROS generation are related to the antiproliferative activity of some representative octahedral cyclometallated platinum(iv) compounds (2b and 2c).


Comprehensive Reactive Receiver Modeling for Diffusive Molecular Communication Systems: Reversible Binding, Molecule Degradation, and Finite Number of Receptors.
This paper studies the problem of receiver modeling in molecular communication systems. We consider the diffusive molecular communication channel between a transmitter nano-machine and a receiver nano-machine in a fluid environment. The information molecules released by the transmitter nano-machine into the environment can degrade in the channel via a first-order degradation reaction and those that reach the receiver nano-machine can participate in a reversible bimolecular reaction with receiver receptor proteins. Thereby, we distinguish between two scenarios. In the first scenario, we assume that the entire surface of the receiver is covered by receptor molecules. We derive a closed-form analytical expression for the expected received signal at the receiver, i.e., the expected number of activated receptors on the surface of the receiver. Then, in the second scenario, we consider the case where the number of receptor molecules is finite and the uniformly distributed receptor molecules cover the receiver surface only partially. We show that the expected received signal for this scenario can be accurately approximated by the expected received signal for the first scenario after appropriately modifying the forward reaction rate constant. The accuracy of the derived analytical results is verified by Brownian motion particle-based simulations of the considered environment, where we also show the impact of the effect of receptor occupancy on the derived analytical results.


Migration timing and its determinants for nocturnal migratory birds during autumn migration.
1. Migration is a common strategy used by birds that breed in seasonal environments, and multiple environmental and biological factors determine the timing of migration. How these factors operate in combination during autumn migration, which is considered to be under weaker time constraints relative to spring migration, is not clear. 2. Here, we examine the patterns and determinants of migration timing for nocturnal migrants during autumn migration in the north-eastern USA using nocturnal reflectivity data from 12 weather surveillance radar stations and modelled diurnal probability of occurrence for 142 species of nocturnal migrants. We first model the capacity of seasonal atmospheric conditions (wind and precipitation) and ecological productivity (vegetation greenness) to predict autumn migration intensity. We then test predictions, formulated under optimal migration theory, on how migration timing should be related to assemblage-level estimates of body size and total migration distance within the context of dietary guild (insectivore and omnivore) and level of dietary plasticity during autumn migration. 3. Our results indicate seasonal declines in ecological productivity delineate the beginning and end of peak migration, whose intensity is best predicted by the velocity of winds at migration altitudes. Insectivorous migrants departed earlier in the season and, consistent with our predictions, large-bodied and long-distance insectivorous migrants departed the earliest. Contrary to our predictions, large-bodied and some long-distance omnivorous migrants departed later in the season, patterns that were replicated in part by insectivorous migrants that displayed dietary plasticity during autumn migration. 4. Our findings indicate migration timing in the region is dictated by optimality strategies, modified based on the breadth and flexibility of migrant's foraging diets, with declining ecological productivity defining possible resource thresholds during which migration occurs when winds at migration altitudes are mild. These observations provide the basis to assess how avian migration strategies may be affected by adjustments in seasonal patterns of atmospheric circulation and ecological productivity that may occur under global climate change.


Production and characterization of specific antibodies for evaluation of glycated insulin in plasma and biological tissues.
Previous studies have shown that glycation of insulin occurs in pancreatic beta-cells under conditions of hyperglycaemia and that the site of glycation is the N-terminal Phe(1) of the insulin B-chain. To enable evaluation of glycated insulin in diabetes, specific antibodies were raised in rabbits and guinea-pigs by using two synthetic peptides (A: Phe-Val-Asn-Gln-His-Leu-Cys-Tyr, and B: Phe-Val-Asn-Gln-His-Leu-Tyr-Lys) modified by N-terminal glycation and corresponding closely to the N-terminal sequence of the glycated human insulin B-chain. For immunization, the glycated peptides were conjugated either to keyhole limpet haemocyanin or ovalbumin using glutaraldehyde, m-maleimidobenzoyl-N-hydroxysuccinimide ester or 1-ethyl-3-(3-dimethylamino propyl) carbodiimide hydrochloride. Antibody titration curves, obtained using I(125)-tyrosylated tracer prepared from glycated peptide A, revealed high-titre antisera in five groups of animals immunized for 8-28 weeks. The highest titres were observed in rabbits and guinea-pigs immunized with peptide B coupled to ovalbumin using glutaraldehyde. Under radioimmunoassay conditions, these antisera exhibited effective dose (median) (ED(50)) values for glycated insulin of 0.3-15 ng/ml and 0.9-2.5 ng/ml respectively, with negligible cross-reactivity against insulin or other islet peptides. The degree of cross-reaction with glycated proinsulin was approximately 50%. Glycated insulin in plasma of control and hydrocortisone-treated diabetic rats measured using rabbit 3 antiserum (1:10 000 dilution; sensitivity <19 pg/ml) was 0. 08+/-0.01 and 1.5+/-0.6 ng/ml (P<0.01), corresponding to 4 and 16% of total circulating insulin concentration respectively. Immunocytochemistry studies of the pancreas of streptozotocin-treated diabetic rats using a 1:1000 dilution of guinea-pig 2 antiserum revealed clusters of fluorescent positively stained cells in islets. These studies document the successful production of polyclonal antisera specific for glycated insulin and their usefulness in radioimmunoassays and immunocytochemistry. The demonstration of glycated insulin in plasma and islets of animal models of diabetes supports the view that glycation of insulin is involved in the pathogenesis of this disease.


A theoretical investigation into the effects of temperature on spatiotemporal dynamics of EET in the FMO complex.
Methodologies are presented in which population dynamics are evolved in the exciton basis and spatiotemporal movement of excitations is subsequently obtained by projection to the site basis. Fluctuations of system eigenstates are explicitly included through vibrations of the chromophores, which are parametrized by ab initio calculations. Two limiting cases of dynamics are considered, namely, the incoherent regime, where state populations correspond to ensembles of classical Landau-Zener (LZ) trajectories, and the coherent regime, where the density matrix is propagated by the quantum Liouville equation (QLE). For QLE simulations, population dynamics show that bacteriochlorophyll a1 and a2 effectively act as a single unit at 77 K but as independent chromophores at 300 K. Population beatings for the lower energy exciton states are considerably slower at physiological temperatures, thus assisting transfer to the sink. Results from LZ trajectories indicate that, within the classical picture, higher temperatures result in a lower probability of the exciton reaching the sink. A broadening of the excitonic spectrum at high temperature alters the pathways of the excitons in the LZ formalism and also increases the possibility of trapping. This study supports the view that a coherent mechanism may assist EET at physiological temperatures since the trapping of excitations in intermediate energy sites is prevented. Furthermore, delocalized vibrations (i.e., superpositions of independent oscillators) are found to assist energy transfer at short times.


Efficacy and Safety of Sofosbuvir/Velpatasvir Plus Ribavirin in Patients with Hepatitis C Virus-Related Decompensated Cirrhosis.
A fixed-dose combination of sofosbuvir/velpatasvir (SOF/VEL) plus weight-based ribavirin (RBV) for 12 weeks is recommended for the treatment of patients with hepatitis C virus (HCV)-associated decompensated cirrhosis. However, large global studies, while confirming the effectiveness of SOF/VEL in a broad range of patients, often exclude these patients. This Phase 2, single-arm, open-label study in adult patients with HCV-associated decompensated cirrhosis in France and the USA aimed to provide further data on the safety and efficacy of SOF/VEL plus RBV for 12 weeks in this population. Patients were treated with a fixed-dose combination of SOF 400 mg/VEL 100 mg plus weight-based RBV once daily for 12 weeks. The inclusion criteria were chronic HCV infection (≥6 months), quantifiable HCV RNA at screening, Child-Turcotte-Pugh class B or C cirrhosis, and liver imaging within 6 months of Day 1 to exclude hepatocellular carcinoma. Among 32 patients who initiated treatment, 78.1% achieved sustained virologic response 12 weeks after the end of treatment (SVR12). Failure to achieve SVR12 was due to non-virologic reasons (investigator discretion, n = 1; death, n = 6). All 25 patients in the per-protocol population achieved SVR12 and all but one achieved sustained virologic response 24 weeks after the end of treatment. Adverse events (AEs) were as expected for a patient population with advanced liver disease. All Grade 3-4 and serious AEs and deaths were deemed unrelated to treatment. In patients with HCV-associated decompensated cirrhosis, SOF/VEL plus RBV achieved high SVR12 rates and was generally well tolerated.


Clinical Translation of Aptamers for COVID-19.
The COVID-19 etiologic agent, SARS-CoV-2, continues to be one of the leading causes of death on a global scale. Although efficient methods for diagnosis and treatment of COVID-19 have been developed, new methods of battling SARS-CoV-2 variants and long COVID are still urgently needed. A number of aptamers have demonstrated tremendous potential to be developed into diagnostic and therapeutic agents for COVID-19. The translation of the aptamers for clinical uses, however, has been extremely slow. Overcoming the difficulties faced by aptamers would advance this technology toward clinical use for COVID-19 and other serious disorders.


Improved detection of urothelial carcinoma in situ with hexaminolevulinate fluorescence cystoscopy.
In this European multicenter study we compared hexaminolevulinate (HAL) fluorescence cystoscopy and standard white light cystoscopy for the detection of carcinoma in situ (CIS) in patients suspected of having high risk bladder cancer. This study was a prospective controlled, within-patient comparison of standard and HAL fluorescence cystoscopy. Eligible patients received an intravesical instillation of 50 ml HAL 8 mM solution. Cystoscopy was performed using a D light system, which provided white and blue light at 375 to 440 nm. The bladder wall was inspected and mapped, first under white light, followed by blue light. All tumors and suspicious areas identified under white light and by red fluorescence were resected or biopsied. Histological findings were assessed by an independent central pathologist blinded to the identity of the biopsies. Of 211 evaluable patients 83 (39%) had CIS, of whom 18 (22%) were detected by HAL cystoscopy only, 62 (75%) were detected by standard and HAL cystoscopy, 2 (2%) were detected by standard cystoscopy only and 1 (1%) was detected by nonguided biopsy. Therefore, HAL cystoscopy identified 28% more patients with CIS than standard cystoscopy. The side effects of HAL instillation were negligible and no unexpected events were reported. HAL fluorescence cystoscopy improves the detection of bladder CIS significantly, which has consequences for clinical management and may improve the patient prognosis. The procedure is easily implemented as an adjunct to standard cystoscopy and it adds no significant risk of complications.


anti-Selective direct catalytic asymmetric Mannich-type reaction of hydroxyketone providing beta-amino alcohols.
An anti-selective direct catalytic asymmetric Mannich-type reaction is described. The Et(2)Zn/(S,S)-linked-BINOL 1 = 4/1 complex promoted a Mannich-type reaction of 2-hydroxy-2'-methoxyacetophenone (2) and N-diphenylphosphinoyl imines 3. Using as little as 0.25-1 mol % of the catalyst, we obtained Mannich adducts 4 in excellent yield (up to 99%), diastereoselectivity (anti/syn = up to >98/2), and enantiomeric excess (up to >99.5%). The anti-selectivity in the present system is complementary to that observed using previously reported methods, providing a novel efficient method to synthesize anti-beta-amino alcohols in a highly enantioselective manner. Facile deprotection of the N-diphenylphosphinoyl group and commercial availability of both Et(2)Zn solution and (S,S)-linked-BINOL 1 also make the present catalysis practical.


Conformity of carotid stents with vascular anatomy: evaluation in carotid models.
Conformity between self-expanding Wallstents and vascular anatomy is limited. Because of a lack of longitudinal flexibility, straightening effects on vascular curves occur and may result in stent-induced kinking. Our purpose was to evaluate the conformity of self-expanding stents with the course and endoluminal surface of silicone models of the normal human carotid artery. Five different types of self-expanding carotid stents were implanted into simplified pulsatile perfused silicone models of the carotid bifurcation. The models embody elastic properties of the vessel wall similar to those of normal human arteries. All stents had the same nominal diameter and length and bridged the external carotid artery origin as well as a consecutive curve at the initial segment of the internal carotid artery. Conventional radiographs of the model were compared before and after stent placement to record changes of shape and course of the silicone artery. Dehiscences between stent filaments and arterial wall were measured on digital subtraction angiograms of the model. Implantation of braided Wallstents or the Expander with continuous filaments induced considerable straightening effects on the bifurcation angle, as well as on the curves of the internal carotid artery. Segmented designs of modular nitinol stents complied better with vascular tortuosity and showed improved adaption between stent and the endoluminal surface of the model. Model experiments show that segmented nitinol stents improve the conformity between the prosthesis and vascular anatomy, and confirm new carotid stent concepts as an alternative to the Wallstent.


Acute and chronic effects of Δ9-tetrahydrocannabinol (THC) on cerebral blood flow: A systematic review.
Acute and chronic exposure to cannabis and its main psychoactive component, Δ9-tetrahydrocannabinol (THC), is associated with changes in brain function and cerebral blood flow (CBF). We therefore sought to systematically review the literature on the effects of THC on CBF following PRISMA guidelines. Studies assessing the acute and chronic effects of THC on CBF, perfusion and volume were searched in the PubMed database between January 1972 and June 2019. We included thirty-four studies, which altogether investigated 1259 humans and 28 animals. Acute and chronic THC exposure have contrasting and regionally specific effects on CBF. While acute THC causes an overall increase in CBF in the anterior cingulate cortex, frontal cortex and insula, in a dose-dependent manner, chronic cannabis use results in an overall reduction in CBF, especially in the prefrontal cortex, which may be reversed upon prolonged abstinence from the drug. Future studies should focus on standardised methodology and longitudinal assessment to strengthen our understanding of the region-specific effects of THC on CBF and its clinical and functional significance.


Pattern Recognition Receptors and Control of Innate Immunity: Role of Nucleic Acids.
The innate immune system protects against infectious microbes by the recognition of pathogen- associated molecular patterns, which serve to detect pathogens on the host cell surface or in endosomes by pattern recognition receptors such as Toll-like receptors, nucleotide-binding oligomerization domain-containing protein-1-like receptors, mannose-receptor, or retinoic acid-inducible gene-1- like receptors that initiate proper host defense mechanisms. In addition to pathogen-associated molecular patterns, a series of endogenous danger-associated molecular patterns, such as nucleic acids, are recognized by pattern recognition receptors, which serve as body´s own alarm signals under sterile conditions, such as ischemic injuries, trauma, tumors, tissue transplants, or autoimmune diseases. Thus, exogenous as well as endogenous nucleic acids can function as "alarmins" to alert the body about danger or disease by triggering inflammation, dendritic cell maturation, and stimulate the immune response resulting in the release of cytokines, which in turn can augment the local inflammatory environment. Moreover, danger-associated molecular patterns such as nucleic acids can act as cofactor in the activation of pattern recognition receptors in situations of cellular stress or upon infection leading to a massive amplification of the inflammatory response. As a consequence, acute and also chronic inflammatory diseases such as rheumatoid arthritis, cancer, or atherosclerosis may depend on such perpetuated proinflammatory responses involving activities of nucleic acids. As antagonists, RNase1 or DNase administration or nucleic acid complexing agents may result in a significant blockade of the outcome of particular pathological situations and in considerable tissue protection.


Simple non-invasive prenatal detection of Hb Bart's disease by analysis of fetal erythrocytes in maternal blood.
To investigate a simple non-invasive technique for early detection of Hemoglobin (Hb) Bart's disease. Maternal blood smears from 8 known Hb Bart's pregnancies and 40 at-risk pregnancies were investigated. Maternal peripheral blood smears were stained with fluorescence-labeled monoclonal antibodies against alpha- and embryonic zeta-globin chains. Fetal nonnucleated red blood cells, stained with anti-zeta but not with anti-alpha globin antibodies were found in 15 out of 16 affected pregnancies but were not detected in 23 out of 24 unaffected pregnancies. Results showed that non-invasive immunofluorescence staining of maternal blood is a feasible approach for screening Hb Bart's disease before ultrasound manifestation in affected pregnancies.


Supplementation of Probiotic Butyricicoccus pullicaecorum Mediates Anticancer Effect on Bladder Urothelial Cells by Regulating Butyrate-Responsive Molecular Signatures.
In bladder cancer, urothelial carcinoma is the most common histologic subtype, accounting for more than 90% of cases. Pathogenic effects due to the dysbiosis of gut microbiota are localized not only in the colon, but also in regulating bladder cancer distally. Butyrate, a short-chain fatty acid produced by gut microbial metabolism, is mainly studied in colon diseases. Therefore, the resolution of the anti-cancer effects of butyrate-producing microbes on bladder urothelial cells and knowledge of the butyrate-responsive molecules must have clinical significance. Here, we demonstrate a correlation between urothelial cancer of the bladder and Butyricicoccus pullicaecorum. This butyrate-producing microbe or their metabolite, butyrate, mediated anti-cancer effects on bladder urothelial cells by regulating cell cycle, cell growth, apoptosis, and gene expression. For example, a tumor suppressor against urothelial cancer of the bladder, bladder cancer-associated protein, was induced in butyrate-treated HT1376 cells, a human urinary bladder cancer cell line. In conclusion, urothelial cancer of the bladder is a significant health problem. To improve the health of bladder urothelial cells, supplementation of B. pullicaecorum may be necessary and can further regulate butyrate-responsive molecular signatures.


Midline Submental Orotracheal Intubation in Maxillofacial Injuries: A Substitute to Tracheostomy Where Postoperative Mechanical Ventilation is not Required.
Maxillofacial fractures present unique airway problems to the anaesthesiologist. Nasotracheal intubation is contraindicated due to associated Lefort I, II or III fractures. The requirement for intraoperative maxillomandibular fixation (MMF) to re-establish dental occlusion in such cases precludes orotracheal intubation. Tracheostomy has a high complication rate and in many patients, an alternative to the oral airway is not required beyond the perioperative period. Hernandez1 in 1986 first described "The submental route for endotracheal intubation". Later some workers faced difficult tube passage, bleeding, and sublingual gland involvement with this approach. They modified this to strict midline submental intubation and there were no operative or postoperative complications in their cases.67&8. Therefore we used mid line approach for submental orotracheal intubation in this study to demonstrate its feasibility and reliability and that it can be used as an excellent substitute to short term tracheostomy. PATIENTS #ENTITYSTARTX00026; We used midline submental intubation in 25 cases selected out of 310 consecutively treated patients with maxillofacial trauma over a 3 year period. After induction orotracheal intubation was done with spiral re-inforced tube. A 1.5-2.0 cm skin incision was made in the submental region in the midline 2.0 cm behind the symphysis and endotracheal tube was taken out through this incision in all the cases. At the end of the surgery the procedure was reversed, the submental wound was stitched; all the patients could be extubated & none of them required post-operative mechanical ventilation. There were no significant operative or postoperative complications. Postoperative submental scarring was acceptable([6]). We conclude that midline submental intubation is a simple and useful technique with low morbidity. It can be chosen in selected cases of maxillofacial trauma and is an excellent substitute to tracheostomy where postoperative mechanical ventilation is not required.


Effect of Mindfulness on Empathy and Self-Compassion: An Adapted MBCT Program on Filipino College Students.
Attending college is meaningful for many young adults. This period is marked by physical, emotional, and psychological changes that can have both positive and negative effects on college students. The last two decades have seen an alarming increase in the number of college students who suffer from mental health conditions, such as depression, suicide, anxiety, and alcohol abuse. It is recommended that actions to support the students' wellbeing must be creative and evidence-based. Research suggests that a mindfulness-based intervention may be an effective strategy to address mental health conditions among college students. This study was done to examine the efficacy of an adapted mindfulness-based cognitive therapy (MBCT) program that was implemented in a classroom setting in the Philippines and to explore how mindfulness practice can affect empathy and self-compassion on senior Filipino college students aged 19-22 years old. Two classes were used to compare the effects of mindfulness intervention. One class underwent the adapted MBCT program while the other class underwent the same kind of class without mindfulness interventions. Self-report measures of the Five Facet Mindfulness Questionnaire, Perspective Taking subscale and Empathic Concern subscale of Interpersonal Reactivity Index, and Self-compassion scale-short form were administered before undergoing the adapted MBCT and after the program. After going through the adapted MBCT, college students' mindfulness significantly improved. Empathy and self-compassion also significantly improved after undergoing the program. This corroborates previous studies done on mindfulness and its efficacy with adolescents and suggests how practicing mindfulness can improve empathy and self-compassion with Filipino college students. It provides a promising groundwork for the emerging interest and research in Asia, particularly in the Philippines, on how the practice of mindfulness can help with the mental health of college students.


Role of non-macrophage cell-derived HMGB1 in oxaliplatin-induced peripheral neuropathy and its prevention by the thrombin/thrombomodulin system in rodents: negative impact of anticoagulants.
Macrophage-derived high mobility group box 1 (HMGB1), a damage-associated molecular pattern (DAMP) protein, plays a key role in the development of chemotherapy-induced peripheral neuropathy (CIPN) caused by paclitaxel in rodents. Endothelial thrombomodulin (TM) promotes thrombin-induced degradation of HMGB1, and TMα, a recombinant human soluble TM, abolishes peripheral HMGB1-induced allodynia in mice. We thus examined whether HMGB1, particularly derived from macrophages, contributes to oxaliplatin-induced neuropathy in mice and analyzed the anti-neuropathic activity of the TM/thrombin system. CIPN models were created by the administration of oxaliplatin in mice and rats, and the nociceptive threshold was assessed by von Frey test or paw pressure test. Macrophage-like RAW264.7 cells were stimulated with oxaliplatin in vitro. Proteins were detected and/or quantified by Western blotting, immunostaining, or enzyme-linked immunosorbent assay. Intraperitoneal administration of an anti-HMGB1-neutralizing antibody (AB) at 1 mg/kg prevented the oxaliplatin-induced allodynia in mice and rats. Antagonists of Toll-like receptor (TLR) 4, receptor for advanced glycation end products (RAGE) and CXCR4 among the HMGB1-targeted pro-nociceptive receptors, also mimicked the anti-neuropathic activity of AB in mice. Macrophage accumulation in the sciatic nerve was observed in mice treated with paclitaxel, but not oxaliplatin, and neither macrophage depletion nor inhibitors of macrophage activation affected oxaliplatin-induced allodynia. Oxaliplatin was 10- to 100-fold less potent than paclitaxel in releasing HMGB1 from macrophage-like RAW264.7 cells. Like AB, TMα at 10 mg/kg prevented the oxaliplatin-induced allodynia in mice as well as rats, an effect abolished by argatroban at 10 mg/kg, a thrombin inhibitor. The anti-neuropathic activity of TMα in oxaliplatin-treated mice was suppressed by oral anticoagulants such as warfarin at 1 mg/kg, dabigatran at 75 mg/kg, and rivaroxaban at 10 mg/kg, but not antiplatelet agents such as aspirin at 50 mg/kg and clopidogrel at 10 mg/kg. Repeated administration of the anticoagulants gradually developed neuropathic allodynia and elevated plasma HMGB1 levels in mice treated with a subeffective dose of oxaliplatin. Our data thus suggests a causative role of HMGB1 derived from non-macrophage cells in oxaliplatin-induced peripheral neuropathy and a thrombin-dependent anti-neuropathic activity of exogenous TMα and, most probably, endogenous TM.


Treatable traits models of care.
Treatable traits is a personalized approach to the management of respiratory disease. The approach involves a multidimensional assessment to understand the traits present in individual patients. Traits are phenotypic and endotypic characteristics that can be identified, are clinically relevant and can be successfully treated by therapy to improve clinical outcomes. Identification of traits is followed by individualized and targeted treatment to those traits. First proposed for the management of asthma and chronic obstructive pulmonary disease (COPD) the approach is recommended in many other areas of respiratory and now immunology medicine. Models of care for treatable traits have been proposed in different diseases and health care setting. In asthma and COPD traits are identified in three domains including pulmonary, extrapulmonary and behavioural/lifestyle/risk-factors. In bronchiectasis and interstitial lung disease, a fourth domain of aetiological traits has been proposed. As the core of treatable traits is personalized and individualized medicine; there are several key aspects to treatable traits models of care that should be considered in the delivery of care. These include person centredness, consideration of patients' values, needs and preferences, health literacy and engagement. We review the models of care that have been proposed and provide guidance on the engagement of patients in this approach to care.


The Nairobi Hospice--tribute to the late professor Edward George Kasili.
Professor Edward George Kasili is regarded as the father of the hospice movement in Kenya. In 1990, he together with Ruth Wooldridge and Jane Moore founded the Nairobi Hospice which takes care of patients with life limiting illnesses from all walks of life. This was of course not easy since even now the mentality of people towards dying is still the same, where people fear talking about death. The hospice movement tries to help somebody die well, die with dignity. The Nairobi Hospice had the privilege of caring for Professor Kasili in his last days before he passed on due to complications of cancer. This article is a tribute to this great man for his forward thinking and for starting the hospice movement.


Success stories of implementation of antimicrobial stewardship: a narrative review.
It has been increasingly recognized that antimicrobial stewardship (AMS) has to be a key component of any efforts that aim to mitigate the current global antimicrobial resistance (AMR) crisis. It has also become evident that AMR is a problem that cannot be tackled by single institutions or physicians, but needs concerted actions at regional, national and supra-national levels. However, it is easy to become discouraged, given the problems that are often encountered when implementing AMS. The aim of this review is to highlight some of the success stories of AMS strategies, and to describe the actions that have been taken, the outcomes that have been obtained, and the obstacles that have been met. Although the best approach to effective AMS remains elusive and may vary significantly among settings, these diverse examples from a range of healthcare contexts demonstrate that effective AMS is possible. Such examples will inform others and encourage them to formally evaluate and share their results with the global stewardship community.


Resolution of cervical dysplasia is associated with T-cell proliferative responses to human papillomavirus type 16 E2.
The 'high-risk' human papillomaviruses (HPVs) cause persistent infections of the anogenital region that may resolve spontaneously following activation of a protective immune response. The aim of this study was to determine whether cell-mediated immunity (CMI) to the early protein E2 was associated with disease regression and to establish whether E2 CMI and antibodies to L1 virus-like particles (VLPs) were associated markers of immunity to HPV. Lymphoproliferative responses to histidine-tagged E2 and antibody responses to VLPs were measured in patients with persistent cervical dysplasia, those whose disease had recently resolved and normal controls. Resolvers had significantly higher E2-specific lymphoproliferative responses when compared with normal controls or persisters, whereas there was no significant difference between the persisters and the normal controls. The T cells stimulated by E2 secreted high levels of gamma interferon (IFN-gamma), consistent with a type 1 helper (Th1) phenotype. VLP IgG responses were associated with current or previous HPV infection, but not with disease regression or a lymphoproliferative response to E2. Major histocompatibility complex class I-restricted T cells secreted IFN-gamma following stimulation with E1, and E2 peptides were detected more frequently in the persister group. The data showed that lymphoproliferative responses to E2 with a cytokine profile indicative of Th1 are associated with disease resolution, supporting the development of a therapeutic vaccine that activates this type of response for the treatment of individuals with pre-existing disease.


Pyrroloquinoline quinone ameliorates renal fibrosis in diabetic nephropathy by inhibiting the pyroptosis pathway in C57BL/6 mice and human kidney 2 cells.
Diabetic nephropathy (DN), which is characterized by renal fibrosis, is a major complication of diabetes, a disease that afflicted more than 460 million people worldwide in 2019. Pyroptosis is an essential signaling pathway in DN-related injuries, such as renal fibrosis. Pyrroloquinoline quinone (PQQ) is a naturally occurring bioactive compound that protects human kidney 2 (HK-2) cells from oxidative stress-induced damage caused by high glucose concentrations. However, the nature and underlying mechanism of the effect of PQQ on DN-related renal fibrosis remains unclear. In this study, we evaluated whether PQQ has potential protective effects against renal fibrosis due to DN by establishing type 1 diabetes in mice via streptozotocin treatment and then inhibiting their pyroptosis signaling pathway. We found that compared to control mice, the area of renal fibrosis and injury were significantly increased in diabetic mice, and this was accompanied by increased levels of expression of collagen Ⅰ and transforming growth factor-β1; increased concentrations of the inflammatory cytokines, interleukin (IL)-1β, IL-6, and tumor necrosis factor-α; and activation of the pyroptosis pathway components nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing 3 (NLRP3), caspase-1, IL-1β, and IL-18. All of these changes were reversed by PQQ treatment. Analogously, we treated cultured HK-2 cells with a high concentration of glucose (35 mmol/L), which caused these cells to exhibit significantly increased concentrations of reactive oxygen species (ROS), phosphorylated (p)-nuclear factor kappa B (NF-κB), p-IkappaB, NLRP3, caspase-1, IL-1β, and IL-18, and the loss of mitochondrial transmembrane potential. However, PQQ treatment significantly blunted these effects. In conclusion, in this study we demonstrated that PQQ attenuates renal fibrosis by alleviating mitochondrial dysfunction, reducing ROS production, and inhibiting the activation of the NF-κB/pyroptosis pathway under conditions of DN and hyperglycemia.


Design and responses of Butterworth and critically damped digital filters.
For many years the Butterworth lowpass filter has been used to smooth many kinds of biomechanical data, despite the fact that it is underdamped and therefore overshoots and/or undershoots data during rapid transitions. A comparison of the conventional Butterworth filter with a critically damped filter shows that the critically damped filter not only removes the undershooting and overshooting, but has a superior rise time during rapid transitions. While analog filters always create phase distortion, both the critically damped and Butterworth filters can be modified to become zero-lag filters when the data are processed in both the forward and reverse directions. In such cases little improvement is realized by applying multiple passes. The Butterworth filter has superior 'roll-off' (attenuation of noise above the cutoff frequency) than the critically damped filter, but by increasing the number of passes of the critically damped filter the same 'roll-off' can be achieved. In summary, the critically damped filter was shown to have superior performance in the time domain than the Butterworth filter, but for data that need to be double differentiated (e.g. displacement data) the Butterworth filter may still be the better choice.


The (pro)renin receptor and soluble (pro)renin receptor in choriocarcinoma.
This study aimed to determine if the (pro)renin receptor (ATP6AP2) changes the cellular profile of choriocarcinomas from cytotrophoblast cells to terminally syncytialised cells and ascertain whether this impacts the invasive potential of choriocarcinoma cells. Additionally, we aimed to confirm that FURIN and/or site 1 protease (MBTPS1) cleave soluble ATP6AP2 (sATP6AP2) in BeWo choriocarcinoma cells and determine whether sATP6AP2 levels reflect the cellular profile of choriocarcinomas. BeWo choriocarcinoma cells were treated with ATP6AP2 siRNA, FURIN siRNA, DEC-RVKR-CMK (to inhibit FURIN activity), or PF 429242 (to inhibit MBTPS1 activity). Cells were also treated with forskolin, to induce syncytialisation, or vehicle and incubated for 48 h before collection of cells and supernatants. Syncytialisation was assessed by measuring hCG secretion (by ELISA) and E-cadherin protein levels (by immunoblot and immunocytochemistry). Cellular invasion was measured using the xCELLigence real-time cell analysis system and secreted sATP6AP2 levels measured by ELISA. Forskolin successfully induced syncytialisation and significantly increased both BeWo choriocarcinoma cell invasion (P < 0.0001) and sATP6AP2 levels (P = 0.02). Treatment with ATP6AP2 siRNA significantly inhibited syncytialisation (decreased hCG secretion (P = 0.005), the percent of nuclei in syncytia (P = 0.05)), forskolin-induced invasion (P = 0.046), and sATP6AP2 levels (P < 0.0001). FURIN siRNA and DEC-RVKR-CMK significantly decreased sATP6AP2 levels (both P < 0.0001). In conclusion, ATP6AP2 is important for syncytialisation of choriocarcinoma cells and thereby limits choriocarcinoma cell invasion. We postulate that sATP6AP2 could be used as a biomarker measuring the invasive potential of choriocarcinomas. Additionally, we confirmed that FURIN, not MBTPS1, cleaves sATP6AP2 in BeWo cells, but other proteases (inhibited by DEC-RVKR-CMK) may also be involved.


IL-10 Prevents apoptosis of brain endothelium during bacteremia.
IL-10-deficient mice infected with the relapsing fever bacterium Borrelia turicatae rapidly succumb to a brain hemorrhage if they are unable to clear peak bacteremia. In this study, we investigated the protective role of IL-10 during relapsing-remitting bacteremia and explored the molecular events involved in the protection of brain endothelium by IL-10. Brain endothelial injury was measured with cytotoxicity and diverse apoptotic assays, whereas the signaling pathway analysis was done by quantitative PCR array. The results showed that severe endothelial cell injury leading to hemorrhage in the brain and other organs occurred in IL-10-deficient mice during relapsing-remitting infection. Human brain microvascular endothelial cells (HBMEC) produced abundant proinflammatory mediators upon exposure to whole bacteria or purified bacterial lipoprotein but did not produce any detectable IL-10. Whole bacteria and purified outer membrane lipoprotein rapidly killed HBMEC by apoptosis in a time- and concentration-dependent manner. Exogenous IL-10 protected HBMEC from apoptosis. HBMEC apoptosis during exposure to a low number of bacteria was associated with downregulation of TNF and TNFAIP3 and upregulation of BAX. In contrast, HBMEC apoptosis during exposure to high concentrations of purified outer membrane lipoprotein was associated with marked upregulation of FAS, FAS ligand, and the adaptor molecules RIPK1 and CFLAR. Exogenous IL-10 reversed all the apoptotic signaling changes induced by whole bacteria or its purified lipoprotein. The results indicate that prominent brain endothelial cell apoptosis occurs during relapsing-remitting bacteremia in the absence of IL-10 and point to a prominent role for bacterial lipoprotein-mediated activation of FAS and caspase-3 in this process.


Plasma and erythrocyte relationship of catecholamines in hemodialysis patients.
The function of the autonomic nervous system is based on reciprocal interaction between the sympathetic and parasympathetic parts, most frequently in the form of antagonistic action on target organs. The main mediators of the sympathetic nervous system in the effectors part are catecholamines (CA), which are involved in various physiological processes. Moreover, CA also has a profound effect on the kidneys, being factors that impact on renal haemodynamics, and have been reported to be altered in pathological disorders, e.g. extracellular volume expression, hypertension and cardiovascular complications. The increased sympathetic nerve activity, at least in part, can explain the raised in plasma CA observed in chronic kidney diseases. Furthermore, plasma CA levels in ureamic patients cannot be considered a reliable index of sympathetic activity, due to existence of many factors which may affect their values. In addition, CA released into the circulation, as one of many substances, may penetrate across the cellular membranes of erytrocytes (RBC). Taking these observations together, the aim of the presented study was to investigate for the first time the plasma and erythrocyte relationship of catecholamines in haemodialysis. The studies were performed among 37 haemodialysed patients who were inhabitants of the Lublin commune. Plasma and intracellular concentration of CA were measured prior to and following haemodialysis by high performance liquid chromatography with electrochemical detection. The results suggest that RBC are able to accumulate CA at the stage of terminal renal failure; in addition, the levels of adrenaline and dopamine in RBC depend on the accumulation of urea in plasma. It was also found that the dynamic changes in concentration of RBC adrenaline are an independent predictor of mortality in haemodialysis patients.


[Clinical inertia and treatment adherence in the management of chronic valvular heart diseases].
Valvular heart diseases are of increasing importance among the general adult population. When compared with other heart diseases, there are few trials in the field of valvular heart disease and randomized clinical trials are particularly scarce. Two sets of guidelines exist: one in the USA and the other in Europe. However, they are not always consistent due to the lack of randomized data and it appears that, frequently, there is a gap between the existing guidelines and their effective application.


Low temperature sensing in tulip (Tulipa gesneriana L.) is mediated through an increased response to auxin.
Tulip (Tulipa gesneriana L.) is a bulbous plant species that requires a period of low temperature for proper growth and flowering. The mechanism of sensing the low temperature period is unknown. The study presented in this paper shows that the essential developmental change in tulip bulbs during cold treatment is an increase in sensitivity to the phytohormone auxin. This is demonstrated using a model system consisting of isolated internodes grown on tissue culture medium containing different combinations of the phytohormones auxin and gibberellin. Using mathematical modelling, equations taken from the field of enzyme kinetics were fitted through the data. By doing so it became apparent that longer periods of low temperature resulted in an increased maximum response at a lower auxin concentration. Besides the cold treatment, gibberellin also enhances the response to auxin in the internodes in this in vitro system. A working model describing the relationship between the cold requirement, gibberellin action and auxin sensitivity is put forward. Possible analogies with other cold-requiring processes such as vernalization and stratification, and the interaction of auxin and gibberellin in the stalk elongation process in other plant species are discussed.


[The surgical treatment of aneurysms of the ascending aorta with associated aortic insufficiency. Total replacement with reimplantation of the coronary arteries].
We reviewed our experience with replacement of the ascending aorta and aortic valve with a composite graft and reimplantation of coronary arteries to the tube graft during 8 years interval from April, 1982, to April 1990. 24 patients underwent repair, the mean age was 49.83 years. Annuloaortic ectasia was the most common indication (58.33%), followed by aortic dissection (acute or chronic). Emergency operation was carried out in nine patients with aortic dissection (37.5%) and elective in 15. The mean duration of cardiopulmonary bypass was 118 +/- 4 minutes and of aortic clamping 83.85 +/- 2 minutes. Hospital mortality was 4.17%, reoperation for hemorrhage was 12.5% and perioperative morbidity for other causes was 34.7%. There were one late death. 20 patients were follow-up with a total of 638 patients-months (two patients excluded with insufficient follow-up and one late death). At last follow-up 14 patients were in functional class I. Eight year actuarial survival for the 24 patients was 91%. We believe that replacement of the ascending aorta and aortic valve with a composite graft and coronary arteries reimplantation to the tube graft is more than one satisfactory alternative to supracoronary graft replacement and aortic valve replacement. It offers the advantage of excluding all abnormal aortic tissue, eliminating the risk for later development of complications in the non excluded disease aorta. It supposes the method of choice for patients with anuloaortic-ectasia, aneurysms of the sinuses of Valsalva with aortic insufficiency, and aortic dissection with proximal affectation of coronary arteries and aortic valve.


Association of Skin Psoriasis and Somatic Comorbidity With the Development of Psychiatric Illness in a Nationwide Swedish Study.
Psoriasis is a complex systemic disease with skin involvement, somatic comorbidity, and psychiatric illness (PI). Although this view of psoriasis is widely accepted, potential synergies within this triad of symptoms have not been adequately investigated. To investigate the independent association of skin psoriasis and somatic comorbidity with the development of PI and to assess whether skin psoriasis and somatic comorbidity act synergistically to produce a risk of PI that is greater than the additive associations. Participants were enrolled between January 2005 and December 2010, in this retrospective matched case-control study using secondary (ie, administrative), population-based registry data from Swedish patients in routine clinical care. The dates of analysis were March 2017 to December 2019. Participants were patients with skin psoriasis and control participants without psoriasis matched on age, sex, and municipality, who were all free of preexisting PI. Presence of skin psoriasis and somatic comorbidity (captured through the Charlson Comorbidity Index and the Elixhauser Comorbidity Index). Risk of PI onset (composite of depression, anxiety, and suicidality) is shown using Kaplan-Meier curves stratified by the presence of skin psoriasis and somatic comorbidity. Adjusted associations of skin psoriasis and somatic comorbidity with the development of PI were analyzed using Cox proportional hazards regression models, including interactions to assess synergistic associations. The 3 components of PI were also assessed individually. A total of 93 239 patients with skin psoriasis (mean [SD] age, 54 [17] years; 47 475 men [51%]) and 1 387 495 control participants (mean [SD] age, 54 [16] years; 702 332 men [51%]) were included in the study. As expected, patients with skin psoriasis were more likely to have somatic comorbidity and PI than control participants. Compared with those without skin psoriasis or somatic comorbidity, patients with psoriasis without somatic comorbidity had a 1.32 times higher risk of PI onset (hazard ratio [HR], 1.32; 95% CI, 1.27-1.36; P < .001), whereas patients with psoriasis with somatic comorbidity had a 2.56 times higher risk of PI onset (HR, 2.56; 95% CI, 2.46-2.66; P < .001). No synergistic associations of skin psoriasis and somatic comorbidity with the development of PI were found (HR, 0.93; 95% CI, 0.81-1.04; P = .21). This study found that somatic comorbidity appeared to alter PI onset even more than skin psoriasis. The observed association of skin psoriasis and somatic comorbidity with the development of PI reinforces the need for proactive, holistic treatment of patients with psoriasis.


Cost of establishing a general dentistry department in a small or medium-sized hospital.
To contribute to future dental healthcare policies, this study compiled data on hospital expenses and follow-ups conducted after a hospital dentistry department was established. In addition, the management status and reports on the utility and challenges of establishing a dentistry department were analyzed. The dentistry department was established through fund raising and inaugurated in May 2009. The depreciation period was set at 7 years, and income and expenditure during the 7 years 8 months after opening were compiled. In total, 17.22 million yen was needed for the dentistry department. The average income from dental care was 21.59 million yen per year, and expenditure amounted to 21.54 million yen per year. The findings indicated that a general dentist able to systemically manage patients was essential in a chronic-care hospital. Moreover, the present findings indicate that if general dentistry consultations were performed without excessive investments, after adjusting for personnel expenses, such an initiative would neither yield considerable income nor produce a substantial deficit. Finally, it is imperative to develop staff who are familiar with the costs and management of hospital dentistry and to increase medical fees for consultations with elderly patients.


Increasing screening intentions for oral and pharyngeal cancer.
Oral and pharyngeal cancer is a serious health threat that goes unnoticed by most people. Increasing screenings for oral and pharyngeal cancer is essential to achieving early detection when the disease is most treatable. We tested the effectiveness of a media campaign designed to increase intentions to seek an oral and pharyngeal cancer screening. We further examined whether concern and knowledge of oral and pharyngeal cancer mediated screening intentions. Participants in the intervention condition received messages on posters, handheld fans, pamphlets, and magnets displayed on the sides of cars or trucks. Participants in the intervention and comparison conditions (N = 1,790) were surveyed prior to and after the intervention. Intervention participants reported greater intentions to seek free oral and pharyngeal cancer screenings. Concern about oral and pharyngeal cancer partially mediated the effect whereas knowledge did not. Our media campaign successfully increased screening intentions by heightening concerns.


Theoretically-Based Emotion Regulation Strategies Using a Mobile App and Wearable Sensor Among Homeless Adolescent Mothers: Acceptability and Feasibility Study.
Many adolescent mothers are parenting young children under highly stressful conditions as they are managing first-time parenthood, poverty, lack of housing, school and work, and challenging peer and familial relationships. Mobile health (mHealth) technology has the potential to intervene at various points in the emotion regulation process of adolescent mothers to provide them support for more adaptive emotional and behavioral regulation in the course of their daily life. The goal of this study was to examine the acceptability, feasibility, use patterns, and mechanisms by which a mobile technology used as an adjunct to in-person, provider-delivered sessions fostered adolescent mothers' adaptive emotion regulation strategies under real-life conditions. Participants (N=49) were enrolled in the intervention condition of a larger pilot study of homeless adolescent mothers living in group-based shelters. The mHealth technology. Calm Mom, consisted of a mobile app and a wrist-worn sensorband for the ambulatory measurement and alerting of increased electrodermal activity (EDA), a physiological measurement of stress. We examined logs of mobile app activity and conducted semistructured qualitative interviews with a subsample (N=10) of participants. Qualitative data analysis was guided by the theoretical frames of the intervention and a technology acceptance model and included an analysis of emerging themes and concepts. Overall, participants indicated that one or more of the elements of Calm Mom supported their ability to effectively regulate their emotions in the course of their daily life in ways that were consonant with the intervention's theoretical model. For many adolescent mothers, the app became an integral tool for managing stress. Due to technical challenges, fewer participants received sensorband alerts; however, those who received alerts reported high levels of acceptability as the technology helped them to identify their emotions and supported them in engaging in more adaptive behaviors during real-life stressful situations with their children, peers, and family members. Calm Mom is a promising technology for providing theoretically driven behavioral intervention strategies during real-life stressful moments among a highly vulnerable population. Future research efforts will involve addressing technology challenges and refining tailoring algorithms for implementation in larger-scale studies.


A blue 4',7-diaminoflavylium cation showing an extended pH range stability.
The introduction of two amine substituents in 4' and 7 positions, leads to the formation of a blue flavylium cation, 7-(N,N'-diethylamino)-2-(9-julolidine)-1-benzopyrilium, which is extremely stable across a wide acidic pH range. The kinetic and thermodynamic constants of the multistate system have been calculated by studying the relaxation kinetics after equilibrium perturbation by addition of base (direct pH jumps) or acid (reverse pH jumps). Except for the cis-chalcone, which is an elusive species, the relative energy levels of the other species could be calculated and a global energy level diagram constructed. The diagram explains that the stability of the diamino compound is due to the high energy level of the hemiketal species, which is difficult to access in acidic medium.


Genetic and immunological host factors associated with susceptibility to HIV-1 infection.
The probability of HIV transmission depends on the interplay of many different factors related to infectiousness of the HIV-infected partner, susceptibility of the HIV-uninfected partner, and biological characteristics of HIV strains. Here, we review recent studies of host immunological and genetic factors which may affect susceptibility to HIV-1 infection. These factors are summarized in Table 1. We propose how to explore biological correlates of susceptibility to HIV-1 infection in epidemiological studies, discuss the strengths and limitations of this research, and address the implications for public health.


Activation and migration of CD8 T cells in the intestinal mucosa.
The activation of CD8 T cells was studied using transgenic mice expressing an MHC class I-restricted OVA-specific TCR (OT-I). Immunization of OT-I mice resulted in activation of lymph node T cells and up-regulation of expression of B220, CD11a, and CD44 and caused a preferential loss of mucosal-type T cells from the periphery. Immunization induced dramatic changes in intestinal lymphocytes, including the appearance of a CD11a high population that resembled peripheral T cells and induced a primary cytolytic response. Activation of adoptively transferred OT-I cells induced migration of CD8 T cells into the lamina propria and intraepithelial lymphocyte compartments. The results suggest that Ag presentation in the intestinal epithelium occurs, and that CD8 cells activated in the periphery can readily enter the mucosa during an ongoing immune response.


[Role of free radicals and reactive nitrogen species in the late complications of diabetes mellitus in rats].
In this time-course study the levels of different reactive species, especially those of nitric oxide and peroxynitrite were determined in streptozotocin-induced diabetic rat tissues at different time-points after the onset of the disease, before the development of histopathological damages. Significantly higher steady state free radical concentrations were found in the liver 3 weeks after the onset of diabetes, compared to age matched control groups. Increased nitric oxide levels in diabetic vasculature and kidney, and its rapid reaction with reactive oxygen species, resulted in high peroxynitrite generation. This suggested the onset of processes characteristic to premature aging of the endothelium. According to the histopathological results, there were no signs of late complications in the tissues up to seven weeks after induction of diabetes. In conclusion, the authors' experimental evidences support the idea of a complex role for nitric oxide, reactive oxygen species and peroxynitrite in the development of early diabetic tissue injury before the evolution of late complications. This study showed for the first time a time-course dependence for changes in nitric oxide production in diabetic tissues compared to age-matched controls at an early stage of the disease. These results suggest that oxidative stress in increased at a very early stage of diabetes and, in particular, that high levels of nitric oxide and peroxynitrite could play a decisive role in the development of late complications in the diabetic vasculature and kidney.


Application of least square method for muscular strength estimation in hand motion recognition using surface EMG.
In this study, we describe the application of least square method for muscular strength estimation in hand motion recognition based on surface electromyogram (SEMG). Although the muscular strength can consider the various evaluation methods, a grasp force is applied as an index to evaluate the muscular strength. Today, SEMG, which is measured from skin surface, is widely used as a control signal for many devices. Because, SEMG is one of the most important biological signal in which the human motion intention is directly reflected. And various devices using SEMG are reported by lots of researchers. Those devices which use SEMG as a control signal, we call them SEMG system. In SEMG system, to achieve high accuracy recognition is an important requirement. Conventionally SEMG system mainly focused on how to achieve this objective. Although it is also important to estimate muscular strength of motions, most of them cannot detect power of muscle. The ability to estimate muscular strength is a very important factor to control the SEMG systems. Thus, our objective of this study is to develop the estimation method for muscular strength by application of least square method, and reflecting the result of measured power to the controlled object. Since it was known that SEMG is formed by physiological variations in the state of muscle fiber membranes, it is thought that it can be related with grasp force. We applied to the least-squares method to construct a relationship between SEMG and grasp force. In order to construct an effective evaluation model, four SEMG measurement locations in consideration of individual difference were decided by the Monte Carlo method.


Interplay between columnar and smectic stability in suspensions of polydisperse colloidal platelets.
The phase behavior of a model suspension of colloidal polydisperse platelets is studied using density-functional theory. Platelets are modelled as parallel rectangular prisms of square section l(2) and height h, with length and height distributions given by different polydispersities δ(l) and δ(h). The model is intended to qualitatively represent experimental colloidal platelet suspensions at high densities with a high degree of orientational ordering. We obtain the phase behavior of the model, including nematic, smectic and columnar phases and its dependence on the two polydispersities δ(l) and δ(h). When δ(l) > δ(h) we observe that the smectic phase stabilises first with respect to the columnar. If δ(h) > δ(l) we observe the opposite behavior. Other more complicated cases occur, e.g. the smectic stabilises from the nematic first but then exists a first-order transition to the columnar phase. Our model assumes plate-rod symmetry, but the regions of stability of smectic and columnar phases are non-symmetric in the δ(l) - δ(h) plane due to the different dimensionality of ordering in the two phases. Microsegregation effects, i.e. different spatial distribution for different sizes within the periodic cell, take place in both phases and, in each case, is more apparent in the variable associated with ordering.


[The formation of electrical excitability in cultured frog embryonic myocytes].
Quantitative changes of sodium and potassium ionic currents were studied in the frog embryonic skeletal myocytes cultures under conditions preventing cell fusion and division. During 7 days of culturing the mean values of the sodium and potassium currents densities were 7- and 1.8-fold increased, resp. In currents clamp records the APs were observed only after 5 days of culturing, and in cells with proportion of sodium and potassium conductances above the critical level.


Toxoplasma Actin Is Required for Efficient Host Cell Invasion.
Apicomplexan parasites actively invade host cells using a mechanism predicted to be powered by a parasite actin-dependent myosin motor. In the model apicomplexan Toxoplasma gondii, inducible knockout of the actin gene, ACT1, was recently demonstrated to limit but not completely abolish invasion. This observation has led to the provocative suggestion that T. gondii possesses alternative, ACT1-independent invasion pathways. Here, we dissected the residual invasive ability of Δact1 parasites. Surprisingly, we were able to detect residual ACT1 protein in inducible Δact1 parasites as long as 5 days after ACT1 deletion. We further found that the longer Δact1 parasites were propagated after ACT1 deletion, the more severe an invasion defect was observed. Both findings are consistent with the quantity of residual ACT1 retained in Δact1 parasites being responsible for their invasive ability. Furthermore, invasion by the Δact1 parasites was also sensitive to the actin polymerization inhibitor cytochalasin D. Finally, there was no clear defect in attachment to host cells or moving junction formation by Δact1 parasites. However, Δact1 parasites often exhibited delayed entry into host cells, suggesting a defect specific to the penetration stage of invasion. Overall, our results support a model where residual ACT1 protein retained in inducible Δact1 parasites facilitates their limited invasive ability and confirm that parasite actin is essential for efficient penetration into host cells during invasion. The prevailing model for apicomplexan invasion has recently been suggested to require major revision, based on studies where core components of the invasion machinery were genetically disrupted using a Cre-Lox-based inducible knockout system. For the myosin component of the motor thought to power invasion, an alternative parasite myosin was recently demonstrated to functionally compensate for loss of the primary myosin involved in invasion. Here, we highlight a second mechanism that can account for the surprising ability of parasites to invade after genetic disruption of core invasion machinery. Specifically, residual actin protein present in inducible knockout parasites appears able to support their limited invasion of host cells. Our results have important implications for the interpretation of the apicomplexan invasion model and also highlight significant considerations when analyzing the phenotypes of inducible knockout parasites generated using Cre-Lox technology.


Dynamics of lipid peroxidation and antioxidion of alpha-tocopherol in membranes.
The dynamics of initiation and inhibition of lipid peroxidation by alpha-tocopherol (alpha-Toc) in membranes were investigated under biological conditions using phosphatidylcholine liposomes. First, I examined how superoxide generated in the bulk water phase is able to induce lipid peroxidation in the inner hydrophobic region of the membrane. Second, I studied the localization of the antioxidant OH group of alpha-Toc in membranes and its lipid radical-trapping dynamics. Third, I investigated how alpha-Toc that is oxidized during radical trapping in membranes is recycled by ascorbic acid (AsA) in the bulk water phase. Finally, I studied the deactivation by alpha-Toc of singlet oxygen ((1)O(2)), which was generated by photoirradiation at the membrane surface, in the hydrophobic membrane inner region, and in bulk water, and measured the (1)O(2) deactivating rate constant of alpha-Toc in membranes considering: the concentration and mobility of alpha-Toc molecule in membranes, especially those of its active OH moiety located at the membrane domains, such as the membrane surface polar zone, inner hydrogen belt, and hydrophobic core, and the dielectric constant reflecting the reactivity of the OH moiety and (1)O(2) in the membrane domains where they interact.


The human primary hepatocyte transcriptome reveals novel insights into atorvastatin and rosuvastatin action.
With particular emphasis on interactions between cholesterol homeostasis and drug metabolism we investigate the transcriptome of human primary hepatocytes treated by two commonly prescribed cholesterol lowering drugs atorvastatin and rosuvastatin and by rifampicin that serves as an outgroup as well as a model substance for induction of nuclear pregnane X receptor. Hepatocytes from human donors have been treated with rosuvastatin, atorvastatin, and rifampicin for 12, 24, and 48 h. Expression profiling with cholesterol and drug metabolism enriched low density Steroltalk cDNA and whole genome Affymetrix HG-U133 Plus 2.0 arrays has been applied. Differential expression (DE) of genes and gene set enrichment analysis of KEGG pathways were performed. Lists of differentially expressed genes and gene sets were cross-compared. Selected genes were confirmed by quantitative real-time PCR. Statins lead to: (a) upregulation of cholesterol-related genes indicating an increased LDL uptake and storage of esterified cholesterol, elevated bile acid/drug export and lower capacity to form HDL; (b) perturbation of genes in glucose and fatty acid homeostasis, influencing acetyl-CoA pools, promoting gluconeogenesis and glucose export; (c) elevated expression of ADIPOR2 suggesting increased sensitivity to adiponectin; (d) perturbations in genes of lipoprotein particle formation, differently for each statin; (e) perturbed expression of many metabolic genes that are directly controlled by nuclear receptors constitutive androstan and/or pregnane X. These data provide a novel global insight into hepatic effects of statins, offering biochemical explanations for higher blood glucose in statin-treated patients, and for drug-induced secondary fatty liver disease.


Exposure to the COVID-19 pandemic environment and generosity.
We report data from an online experiment which allows us to study how generosity changed over a 6-day period during the initial explosive growth of the COVID-19 pandemic in Andalusia, Spain, while the country was under a strict lockdown. Participants (n = 969) could donate a fraction of a €100 prize to an unknown charity. Our data are particularly rich in the age distribution and we complement them with daily public information about COVID-19-related deaths, infections and hospital admissions. We find correlational evidence that donations decreased in the period under study, particularly among older individuals. Our analysis of the mechanisms behind the detected decrease in generosity suggests that expectations about others' behaviour, perceived mortality risk and (alarming) information play a key-but independent-role for behavioural adaptation. These results indicate that social behaviour is quickly adjusted in response to the pandemic environment, possibly reflecting some form of selective prosociality.


Electrochemical Immunosensing Platform for the Determination of the 20S Proteasome Using an Aminophenylboronic/Poly-indole-6-carboxylic Acid-Modified Electrode.
The first electrochemical immunosensor for the determination of the 20S proteasome (P20S) was developed, entailing the immobilization of an antibody on an aminophenylboronic/poly-indole-6-carboxylic acid-modified electrode. The proposed electrochemical bioplatform is a simple and feasible analytical tool applicable for the determination of P20S in human plasma, considering its high clinical and biological relevance. Cyclic voltammetry, electrochemical impedance spectroscopy, and square wave voltammetry (SWV) were used to determine the optimal step-by-step process to obtain the electrochemical immunosensor. The interaction of P20S with the recognition layer of the immobilized antibody on the nanostructured surface took place by incubating the electrode in a P20S solution at 20 °C for 2 h. Using SWV as an electro-analytical technique, this immunosensor can quantify P20S. The current was linear with the P20S concentration within two dynamic concentration ranges from 20.0 to 80.0 and 80.0 to 200.0 ng·mL-1 (r2 = 0.992 and 0.98, respectively) with a limit of detection and quantification of 6 and 18 ng·mL-1, respectively. Moreover, the immunosensor showed considerable repeatability and reproducibility, when the determination was done in human serum, which confirms that it is a promising alternative for direct detection of P20S in biological fluids with minimal interference.


IL-21 induces both rapid maturation of human CD34+ cell precursors towards NK cells and acquisition of surface killer Ig-like receptors.
The NK cell maturation from CD34(+) Lin(-) hematopoietic cell precursors is a complex process that requires the direct contact with stromal cells and/or the synergistic effect of different cytokines. In this study we show that IL-21 is capable of inducing an accelerated NK cell maturation when added to cultures of CD34(+) Lin(-) cells isolated from human cord blood supplemented with IL-15, Flt3-L and SCF. After 25 days of culture, 50% of CD56(+) cells expressed various NK cell markers including the NKp46 and NKp30 triggering receptors, the CD94/NKG2A inhibitory receptor and CD16. At day 35, substantial fractions of NK cells expressed KIR, CD8 and CD2, i.e. surface markers expressed by mature NK cells, that are virtually undetectable in developing NK cells cultured in the absence of IL-21. Remarkably, similar to mature NK cells all these markers were included in the CD56(dim) cell fraction, while the CD56(bright) population was only composed of CD94/NKG2A(-) and CD94/NKG2A(+) cells. Thus, IL-21 allows the induction of a full NK cell maturation in vitro and offers an important tool for dissecting the molecular mechanisms involved in different steps of NK cell maturation and in the acquisition of a mature KIR repertoire.


Interaction specificity and coexpression of rice NPR1 homologs 1 and 3 (NH1 and NH3), TGA transcription factors and Negative Regulator of Resistance (NRR) proteins.
The nonexpressor of pathogenesis-related genes 1, NPR1 (also known as NIM1 and SAI1), is a key regulator of SA-mediated systemic acquired resistance (SAR) in Arabidopsis. In rice, the NPR1 homolog 1 (NH1) interacts with TGA transcriptional regulators and the Negative Regulator of Resistance (NRR) protein to modulate the SAR response. Though five NPR1 homologs (NHs) have been identified in rice, only NH1 and NH3 enhance immunity when overexpressed. To understand why NH1 and NH3, but not NH2, NH4, or NH5, contribute to the rice immune response, we screened TGA transcription factors and NRR-like proteins for interactions specific to NH1 and NH3. We also examined their co-expression patterns using publicly available microarray data. We tested five NHs, four NRR homologs (RHs), and 13 rice TGA proteins for pair-wise protein interactions using yeast two-hybrid (Y2H) and split YFP assays. A survey of 331 inter-family interactions revealed a broad, complex protein interaction network. To investigate preferred interaction partners when all three families of proteins were present, we performed a bridged split YFP assay employing YFPN-fused TGA, YFPC-fused RH, and NH proteins without YFP fusions. We found 64 tertiary interactions mediated by NH family members among the 120 sets we examined. In the yeast two-hybrid assay, each NH protein was capable of interacting with most TGA and RH proteins. In the split YFP assay, NH1 was the most prevalent interactor of TGA and RH proteins, NH3 ranked the second, and NH4 ranked the third. Based on their interaction with TGA proteins, NH proteins can be divided into two subfamilies: NH1, NH2, and NH3 in one family and NH4 and NH5 in the other.In addition to evidence of overlap in interaction partners, co-expression analyses of microarray data suggest a correlation between NH1 and NH3 expression patterns, supporting their common role in rice immunity. However, NH3 is very tightly co-expressed with RH1 and RH2, while NH1 is strongly, inversely co-expressed with RH proteins, representing a difference between NH1 and NH3 expression patterns. Our genome-wide surveys reveal that each rice NH protein can partner with many rice TGA and RH proteins and that each NH protein prefers specific interaction partners. NH1 and NH3 are capable of interacting strongly with most rice TGA and RH proteins, whereas NH2, NH4, and NH5 have weaker, limited interaction with TGA and RH proteins in rice cells. We have identified rTGA2.1, rTGA2.2, rTGA2.3, rLG2, TGAL2 and TGAL4 proteins as the preferred partners of NH1 and NH3, but not NH2, NH4, or NH5. These TGA proteins may play an important role in NH1- and NH3-mediated immune responses. In contrast, NH4 and NH5 preferentially interact with TGAL5, TGAL7, TGAL8 and TGAL9, which are predicted to be involved in plant development.


Heterogeneity of lymphocyte subpopulations in severe combined immunodeficiency. Evidence against a stem cell defect.
Surface markers typical of T and B lymphocytes were present on varying proportions of peripheral blood lymphocytes from three infants with severe combined immunodeficiency disease. Despite this, functions mediated by T and B cells were either absent or very minimal in all three, including cell-mediated responses in vivo; the in vitro proliferative response to mitogens, allogeneic cells, or antigens; effector cell function in lymphocyte-antibody lymphocytolytic interaction assays; and in vitro synthesis of IgG, IgA, and IgM. In contrast, mononuclear cells from one of the infants were tested and found capable of lysing both human and chicken antibody-coated erythrocyte targets normally. Co-cultivation experiments with unrelated normal control lymphocytes failed to demonstrate suppressor cell activity for immunoglobulin synthesis in these infants. Augmentations of immunoglobulin production from 310 to 560% over that expected on the basis of individual culture data were noted in co-cultures of one of the infants' cells with two different unrelated normal control cells. These findings suggest that that infant may have had a T helper cell defect or that his T cells were unable to produce soluble factors necessary for B cell differentiation. The finding of cells with differentiation markers characteristic of T and B lymphocytes in each of these patients, though in variable quantities, is further evidence for heterogeneity among patients with the clinical syndrome of severe combined immunodeficiency and argues against the concept that their immunodeficiency was due to a stem cell defect.


Analysis of head acceleration events in collegiate-level American football: A combination of qualitative video analysis and in-vivo head kinematic measurement.
The contact nature of American football has made head acceleration exposure a concern. We aimed to quantify the head kinematics associated with direct helmet contact and inertial head loading events in collegiate-level American football. A cohort of collegiate-level players were equipped with instrumented mouthguards synchronised with time-stamped multiple camera-view video footage of matches and practice. Video-verified contact events were identified as direct helmet contact or inertial head loading events and categorised as blocking, blocked, tackling, tackled or ground contact. Linear mixed-effects models were utilised to compare peak head kinematics between contact event categories. The timestamp-based cross-verification of the video analysis and instrumented mouthguard approach resulted in 200 and 328 direct helmet contact and inertial head loading cases, respectively. Median linear acceleration, angular acceleration and angular velocity for inertial head loading cases was greater than direct helmet contact events by 8% (p = 0.007), 55% (p < 0.001) and 4% (p = 0.007), respectively. Median head kinematics for all contact event categories appeared similar with no pairwise comparison resulting in statistical significance (p > 0.05). The study highlights the potential of combining qualitative video analysis with in-vivo head kinematics measurements. The findings suggest that a number of direct helmet contact events sustained in American football are of lower magnitude to what is sustained during regular play (i.e. from inertial head loading). Additionally, the findings illustrate the importance of including all contact events, including direct helmet contact and inertial head loading cases, when assessing head acceleration exposure and player load during a season of American football.