Porphyromonas gingivalis FimA and Mfa1 fimbriae: Current insights on localization, function, biogenesis, and genotype.
In general, the periodontal pathogen Porphyromonas gingivalis expresses distinct FimA and Mfa1 fimbriae. Each of these consists of five FimA-E and five Mfa1-5 proteins encoded by the fim and mfa gene clusters, respectively. The main shaft portion comprises FimA and Mfa1, whereas FimB and Mfa2 are localized on the basal portion and function as anchors and elongation terminators. FimC-E and Mfa3-5 participate in the assembly of an accessory protein complex on the tips of each fimbria. Hence, they serve as ligands for the receptors on host cells and other oral bacterial species. The crystal structures of FimA and Mfa1 fimbrial proteins were recently elucidated and new insights into the localization, function, and biogenesis of these proteins have been reported. Several studies indicated a correlation between P. gingivalis pathogenicity and the fimA genotype but not the mfa1 genotype. We recently revealed polymorphisms of all genes in the fim and mfa gene clusters. Intriguingly, mfa5 occurred in numerous different forms and underwent duplication. Detailed structural and functional knowledge of the fimbrial proteins in the context of the entire filament could facilitate the development of innovative therapeutic strategies for structure-based drug design.


A case of drug-induced parkinsonism and tardive akathisia with e1143g polymerase γ mutation-innocent bystander or a culprit?
Polymerase γ (POLG) is a protein that plays a pivotal role in the replication of the mitochondrial genome. POLG-related disorders constitute a sequence of overlying phenotypes that can present from early infancy to late adulthood. Parkinsonism is the most common movement disorder associated with POLG mutation. We also summarize all reported cases of POLG-related Parkinsonism, along with a literature review. We present the case of an 80-year-old male presented with complaints of episodic confusion, tremors, and restlessness. He has been on risperidone for psychosis. A normal DaT scan ruled out Parkinson's disease, and molecular analysis for POLG was positive (E1143G). He was diagnosed with drug-induced Parkinsonism and tardive akathisia with an incidental POLG mutation. A literature search revealed 55 cases of "POLG-related Parkinsonism" that met our criteria. These mutations can clinically affect multiple organ systems. Parkinsonism is the most common movement disorder reported in association with POLG mutations. We conducted a molecular analysis in our patient due to his Parkinsonism and recurrent episodes of encephalopathy. E1143G mutation found in our case was coincidental and reported a non-pathogenic or benign variant in literature. E1143G is a non-pathogenic variant and multiple studies have shown that its co-occurrence with other POLG mutations can aggravate disease occurrence and severity. Literature findings and the experience from our own case indicate that the pathogenicity of E1143G is debatable, and future studies involving this particular variant may help understand its pathological consequences.


Intracranial suppuration complicating tropical pyomyositis. Report of two cases.
Tropical pyomyositis complicated by intracranial abscesses is described in two Nigerian male patients, aged five and 50 years. In both cases, the lesions were verified at autopsy. The occurrence of brain abscesses in pyomyositis is briefly reviewed. The complication may not be as rare as previously thought and this will appear if port-mortem examination is performed on most, possibly all, fatal cases of pyomyositis.


Evaluation of the Sensitivity and Specificity of the New Clinical Diagnostic and Classification Criteria for Kashin-Beck Disease, an Endemic Osteoarthritis, in China.
This study aimed to evaluate the sensitivity and specificity of the new clinical diagnostic and classification criteria for Kashin-Beck disease (KBD) using six clinical markers: flexion of the distal part of fingers, deformed fingers, enlarged finger joints, shortened fingers, squat down, and dwarfism. One-third of the total population in Linyou County was sampled by stratified random sampling. The survey included baseline characteristics and clinical diagnoses, and the sensitivity and specificity of the new criteria was evaluated. We identified 3,459 KBD patients, of which 69 had early stage KBD, 1,952 had stage I, 1,132 had stage II, and 306 had stage III. A screening test classified enlarged finger joints as stage I KBD, with a sensitivity and specificity of 0.978 and 0.045, respectively. Shortened fingers were classified as stage II KBD, with a sensitivity and specificity of 0.969 and 0.844, respectively, and dwarfism was classified as stage III KBD with a sensitivity and specificity of 0.951 and 0.992, respectively. Serial screening test revealed that the new clinical classification of KBD classified stages I, II, and III KBD with sensitivities of 0.949, 0.945, and 0.925 and specificities of 0.967, 0.970, and 0.993, respectively. The screening tests revealed that enlarged finger joints, shortened fingers, and dwarfism were appropriate markers for the clinical diagnosis and classification of KBD with high sensitivity and specificity.


Microfluidic pore model study of precipitates induced by the pore-scale mixing of an iron sulfate solution with simulated groundwater.
Precipitates induced by the pore-scale mixing of iron sulfate solutions with simulated groundwater were investigated using a microfluidic pore model to assess the environmental impacts of the infiltration of acid mine drainage into a shallow aquifer. This model was employed to visualize the formation of precipitates in a porous network and to evaluate their physicochemical influences on pore flow. Four types of groundwater (Na-HCO3, Na-SO4, Na-Cl, and Ca-Cl) were evaluated, and precipitation rates were calculated by processing images of precipitates in the pores captured via microscopy. The results showed that all groundwater types formed a yellow-brownish precipitate at the interface of the iron solution and simulated groundwater flow. Microscopic X-ray analyses demonstrated that precipitate morphology varied with groundwater type. Faster precipitation was observed in the following order by groundwater type: Na-HCO3 > Na-Cl > Na-SO4 > Ca-Cl, which was attributed to the different stability constants of the major anions in each simulated groundwater with Fe ions. Chemical equilibrium models suggested that precipitates were Fe minerals, with FeOOH as the predominant form consistent with the results of X-ray photoelectron spectrometry. The presence of FeOOH implies that precipitates may serve as an effective sorption barrier against some nutrients and heavy metals for the underlying groundwater. However, dye-flow experiments suggested that the precipitates may clog aquifer pores, thereby altering hydrogeological properties in the aquifer.


Inhibition of an identified snail postsynaptic neuron evoked by stimulation of a peptidergic interneuron initiating bursting pacemaker activity in another neuron.
Beating neuron V8, whose activity is abolished upon interneuronal stimulation, has been found and identified in the CNS of the snail, Helix pomatia. It was found earlier that stimulation of the same interneuron initiates bursting pacemaker activity in another neuron. Under voltage clamp conditions interneuronal stimulation evoked a slow outward current in the V8 neuron with a latency of about 2 s. Intracellular injection of tetraethylammonium or Cs+ ions into the interneuron greatly increased the slow outward current amplitude. External application of CdCl2 (1 mM), LaCl3 (5 mM) or replacement of Ca2+ by Mg2+ ions reversibly abolished transmission between the interneuron and the V8 neuron. The neuronal membrane conductance increased by about three times on the maximum of the slow outward current development. Computer fitting showed that a decay of the slow outward current can be approximated by the sum of two exponentially decaying components with time constants of about 1 s and 16 s. Extrapolated reversal potential for a fast (1 s) component of the current was about -70 mV. A slow (16 s) component asymptotically decreased upon hyperpolarization and it was observed at even more negative potential than Ek. Theophylline (1 mM) and 3-isobutyl-1-methylxanthine (0.1 mM) reversibly increased the slow outward current amplitude, whereas imidazole (5 mM) and tolbutamide (5 mM) caused its reversible decrease. It is concluded that inhibitory monosynaptic transmission exists between the interneuron and the beating V8 neuron. Interneuronal simulation evokes an increase in the membrane potassium conductance and probably a decrease in stationary potential-dependent sodium conductance of the postsynaptic V8 neuron. The cellular cyclase system seems to be involved in the postsynaptic response.


The chemical composition of the aerial parts of Stachys spreitzenhoferi (Lamiaceae) growing in Kythira Island (Greece), and their antioxidant, antimicrobial, and antiproliferative properties.
The Stachys L. genus has been used in traditional medicine to treat skin inflammations, stomach disorders, and stress. The aim of this study was to investigate the chemical profile and biological activity of the methanolic extract of Stachys spreitzenhoferi Heldr. (Lamiaceae) aerial parts, collected on the island of Kythira, South Greece. The analysis by liquid chromatography coupled with electrospray ionization and high-resolution mass spectrometry [LC-(-)ESI/HRMSn] of the methanol extract revealed the occurrence of thirty-six compounds - flavonoids, phenylethanoid glycosides, iridoids, quinic acid derivatives, aliphatic alcohol glycosides, and oligosaccharides - highlighting the substantial presence, as main peaks, of the iridoid melittoside (2) along with flavonoid compounds such as 4'-O-methylisoscutellarein mono-acetyl-diglycoside/chrysoeriol mono-acetyl-diglycoside (24), trimethoxy- (35) and tetramethoxyflavones (36). This extract was tested for its antimicrobial properties against Gram-positive and negative pathogenic strains. The extract was not active against Gram-negative bacteria tested, but it possessed a good dose-dependent antimicrobial activity towards S. aureus (MIC: 1.0 mg/mL) and L. monocytogenes (MIC: 1.0 mg/mL) Gram-(+) strains. Furthermore, this extract has been tested for its possible antioxidant activity in vitro. In particular, it has been shown that these molecules cause a decrease in DPPH, ABTS, and H2O2 radicals. The extract of S. spreitzenhoferi exhibited anti-DPPH activity (IC50: 0.17 mg/mL), anti-H2O2 activity (IC50: 0.125 mg/mL), and promising antiradical effect with an IC50 value of 0.18 mg/mL for anti-ABTS activity. S. spreitzenhoferi extract caused a decrease in ROS (at the concentration of 200 μg/mL) and an increase in the activity of the antioxidant enzymes SOD, CAT, and GPX in OZ-stimulated PMNs. Furthermore, it exhibited antiproliferative activity against acute myeloid leukemia (U937 cell), causing 50% of cell death at the 0.75 mg/mL.


The effect of posterior vitreous detachment on aflibercept response in diabetic macular oedema.
To investigate the effect of posterior vitreous detachment (PVD) on aflibercept response in treatment-naïve diabetic macular oedema (DME). A retrospective cohort study. One hundred and fifty eyes of 150 treatment-naïve patients with DME were enrolled. The patients were divided into three groups according to their PVD status: group 1 (no PVD during injections), group 2 (PVD during injections) and group 3 (PVD already present initially). Three consecutive aflibercept injections at intervals of 1 month were applied to all patients. The efficacy of the aflibercept treatment on DME was assessed by comparing changes in central retinal thickness (CRT) and best-corrected visual acuity (BCVA) values after three loading dose injections. After three consecutive injections, the mean reduction of CRT in groups 1, 2 and 3 were -199.6±106.9, -224.9±124.1 and -210.7±126.3, respectively. The decrease in CRT was significant in all groups (p<0.001 in all groups, paired-samples t-test); however, mean changes in CRT were similar among groups (all p>0.05, one-way analysis of variance (ANOVA)). The mean improvement in BCVA in groups 1, 2 and 3 was 6.6±0.9, 6.5±0.8 and 6.1±0.4 ETDRS letters, respectively. The improvement of BCVA was significant in all groups (p<0.001 in all groups, paired-samples t-test) and mean changes were similar between groups (p>0.05, one-way ANOVA). There was no significant relationship between the presence or development of PVD and the mean decrease of CRT and improvement of BCVA (for CRT r=-0.052, p=0.531 and for BCVA r=-0.078, p=0.342). In the present study, it was observed that the efficacy of aflibercept treatment in patients with DME did not depend on PVD status.


Genomic loss of imprinting in first-trimester human placenta.
The purpose of this study was to investigate imprinting patterns in first-trimester human placentas. Using samples of 17 first-trimester and 14 term placentas from uncomplicated pregnancies, we assessed loss of imprinting (LOI) at the RNA level in a panel of 14 genes that are known to be imprinted in the placenta with the use of a quantitative allele-specific reverse transcriptase polymerase chain reaction analysis of those genes that contained readout single nucleotide polymorphisms in their transcripts. There is significant LOI (ie, biallelic expression) in all 14 genes in first-trimester placentas. LOI was more variable and generally at lower levels at term. Although there is little difference in gene expression, the level of LOI is higher in the first-trimester placentas, compared with term placentas. Genomic imprinting appears to be a dynamic maturational process across gestation in human placenta. In contrast with prevailing theories, epigenetic imprints may continue to evolve past 12 weeks of gestation.


Fate-mapping of the epithelial seam during palatal fusion rules out epithelial-mesenchymal transformation.
During palatogenesis, fusion of the palatine shelves is a crucial event, the failure of which results in the birth defect, cleft palate. The fate of the midline epithelial seam (MES), which develops transiently upon contact of the two palatine shelves, is still strongly debated. Three major mechanisms underlying the regression of the MES upon palatal fusion have been proposed: (1) apoptosis has been evidenced by morphological and molecular criteria; (2) epithelial-mesenchymal transformation has been suggested based on ultrastructural and lipophilic dye cell labeling observations; and (3) migration of MES cells toward the oral and nasal areas has been proposed following lipophilic dye cell labeling. To verify whether epithelial-mesenchymal transformation of MES cells takes place during murine palatal fusion, we used the Cre/lox system to genetically mark Sonic hedgehog- and Keratin-14-expressing palatal epithelial cells and to identify their fate in vivo. Our analyses provide conclusive evidence that rules out the occurrence of epithelial-mesenchymal transformation of MES cells.


Comparative Evaluation of the Effect of EDTA, EDTAC, NaOCl and MTAD on Microhardness of Human Dentin - An In-vitro Study.
The aim of this study was to evaluate the microhardness of human dentin by using four irrigating solutions. Materials and Methodos: A total of 40 extracted mandibular premolars were selected and sectioned horizontally in the middle third of the root. Forty specimens of 4 mm thickness were embedded in acrylic resin and polished. Four test groups, each group containing ten specimens were immersed in respective irrigating solution and subjected to vicker's microhardness test at T0, T2 and T5min. The data obtained were analyzed using the one way ANOVA followed by Tukey HSD method with ap=0.05 as the level for statistical significance. Suggested that there was no statistically significant difference in mean values between four experimental irrigating solutions. Mixture of Tetracycline isomer i.e. Doxycycline, Citric acid and a Detergent (Tween 80) MTAD not altered the microhardness of root canal dentin significantly and seems to be an appropriate irrigating solution, because of its harmless effect on the microhardness of the root canal dentin.


Solubilization of the picrotoxinin binding receptor from mammalian brain.
The binding sites for alpha-dihydropicrotoxinin (DHP), which is a ligand for the picrotoxin-sensitive component at the benzodiazepine-gamma-aminobutyric acid-receptor-ionophore complex, has been solubilized from rat brain, using 1% Lubrol. A new assay, which involves precipitation of the [3H]DHP-soluble protein complex by gamma-globulin and polyethylene glycol (PEG), followed by centrifugation, is described. The solubilized material bound DHP to two sites with apparent affinities of 0.038 and 1.85 microM. The binding of DHP to the solubilized receptors was inhibited by convulsant and depressant drugs with potencies similar to those required for membrane receptors. The ability of barbiturates to inhibit DHP binding to both solubilized and membrane receptors strongly suggests that barbiturates may interact with the picrotoxin binding component. These data suggest that ligand recognition properties of the picrotoxinin binding are not altered by solubilization. The binding was abolished by urea and partially destroyed by heating the soluble extract at 65 degrees C for 30 min. This new method of measuring the binding of ligands to the solubilized receptors by PEG centrifugation might be used successfully in other solubilization studies.


HBsAg as an important predictor of HBeAg seroconversion following antiviral treatment for HBeAg-positive chronic hepatitis B patients.
Serum quantitative hepatitis B surface antigen (HBsAg) levels may be an important predictor of hepatitis B e antigen (HBeAg) seroconversion (SC) in HBeAg-positive chronic hepatitis B (CHB) patients during antiviral treatment. The pattern of HBsAg variation in CHB patients either with or without SC following tenofovir disoproxil fumarate (TDF) treatment is not clearly understood. Twenty patients with full experimental data were enrolled, and liver biochemistry, serum HBV DNA, and circulating CD4+CD25+ regulatory T cell (Treg) levels were determined at baseline and every 12 weeks after the initiation of TDF treatment (for a total of 96 weeks). In addition, the relationship between HBsAg or HBeAg and alanine aminotransferase (ALT), HBV DNA and Treg levels in SC and non-SC patients was analyzed. In all, 9 patients had undergone HBeAg seroconversion by week 72 of TDF treatment, and biochemical and virological indexes and Treg percentages declined to normal levels. Furthermore, the positive correlation between HBsAg and ALT, HBV DNA and Treg levels was significant for SC patients, but not for non-SC patients. However, for HBeAg, significant positive correlations were or not observed for both SC and non-SC patients. The quantitation of HBsAg is a more useful indicator than HBeAg for distinguishing SC and non-SC patients during TDF treatment. Moreover, HBsAg may be related to immune regulatory property of CHB patients during antiviral treatment.


Percutaneous image-guided screws meditated osteosynthesis of impeding and pathological/insufficiency fractures of the femoral neck in non-surgical cancer patients.
To present percutaneous image-guided screw-mediated osteosynthesis (PIGSMO) for fixation of impending fractures (ImF) and non-displaced/mildly displaced pathological/insufficient fractures (PF/InF) of the femoral neck in non-surgical cancer patients. This is a double-centre single-arm observational study. Retrospective review of electronic records identified all oncologic patients who had undergone femoral neck PIGSMO. Inclusion criteria were: non-displaced or mildly displaced PF/InF, and ImF (Mirels' score ≥8); life expectancy ≥1 month; unsuitability for surgical treatment due to sub-optimal clinical fitness, refusal of consent, or unacceptable delay to systemic therapy. Eleven patients were treated (mean age 63.7±13.5 years) due to ImF (63.6%, mean Mirels' score 10.1), PF (27.3%) or post-radiation InF (9.1%) under CT/fluoroscopy- (36.4%) or CBCT- (63.6%) guidance. Thirty-two screws were implanted and cement injection was added in 36.4% cases. Technical success was 90.9%. No procedure related complications were noted. At 1-month clinical follow-up (pain/walking impairment), 63.6% and 27.3% patients reported significant and mild improvement, respectively. Imaging follow-up (available in 63.6% cases) showed no signs of secondary fractures, neither of screws loosening at mean 2.8 months. Five patients (45.5%) died after PIGSMO (mean time interval 3.6 months). PIGSMO is technically feasible and safe in cancer patients with limited life expectancy; it offers good short-term results. Further prospective studies are required to corroborate mid- and to prove long-term efficacy of the technique.


Home enteral tube feeding in patients with inherited metabolic disorders: safety issues.
Many children with inherited metabolic disorders (IMD), at risk of hypoglycaemia and metabolic decompensation, are dependent on long-term home overnight enteral tube feeding but its safety issues have not been evaluated. To identify common safety issues and carer pressures for patients with IMD on home enteral tube feeds (HETF). Thirty-four patients (53% male; median age 4.1, range: 1.2-15.8 years), with IMD on home continuous overnight tube feeds were recruited. They were all following specialized feeding regimens. A questionnaire, administered by face-to-face interview with carers identified family members involved in feeding, training they received; child safety issues; equipment reliability and carer night time disturbance. The principal problems were: carer sleep disturbance (100%); tube entanglement (71%); untrained secondary carers (71%); faulty pumps (50%); tube blockages (45%); faulty equipment (32%); and child tampering with pumps and feeding equipment (29%). Significant risks for children on HETF with IMD were identified, potentially leading to metabolic decompensation and hospitalization. The safety of feeding equipment, lack of training of extended family members and practical support for carers requires urgent attention.


Diffusion properties of the fornix assessed by deterministic tractography shows age, sex, volume, cognitive, hemispheric, and twin relationships in young adults from the Human Connectome Project.
The fornix is the primary efferent pathway of the hippocampus and plays a central role in memory circuitry. Diffusion tensor imaging has shown changes in the fornix with typical development and aging. Here, the fornix was investigated in 903 healthy young adult participants aged 22-36 years old from the high-spatial resolution 1.25 mm isotropic Human Connectome Project (HCP) diffusion dataset. Manual deterministic tractography was used to assess relationships between fornix diffusion parameters and age, sex, laterality, hippocampus volume, memory scores, and genetic effects in a subgroup of mono- and dizygotic twins. Fornix diffusion metrics were weakly correlated with age over the given age span. While significant hemispheric and sex differences were observed (greater fractional anisotropy (FA) and volume in the right hemisphere; greater FA and volume in females), there was great overlap between the groups. Hippocampus volume measurements showed greater volume in the right hemisphere, were found to be larger in males, and were weakly correlated with fornix FA and volume. Interestingly, all fornix diffusion measurements correlated strongly with fornix volume, suggesting the presence of partial volume effects despite the high-spatial resolution of the data. Both fornix diffusion parameters and hippocampal volumes were able to explain some variance (0.6-5.5%) in the memory tests evaluated. The fornix diffusion parameters were influenced by both genetic and shared environmental factors, displaying greater variability in dizygotic than in monozygotic twins. These findings provide a comprehensive depiction of the fornix in healthy, young individuals, upon which future typical development/aging and pathological studies could anchor.


Novel quinazoline-based EGFR kinase inhibitors: A review focussing on SAR and molecular docking studies (2015-2019).
The over expression of EGFR has been recognized as the driver mechanism in the occurrence and progression of carcinomas such as lung cancer, breast cancer, pancreatic cancer, etcetera. EGFR receptor was thus established as an important target for the management of solid tumors. The occurrence of resistance caused as a result of mutations in EGFR has presented a formidable challenge in the discovery of novel inhibitors of EGFR. This has resulted in the development of three generations of EGFR TKIs. Newer mutations like C797S cause failure of Osimertinib and other EGFR TKIs belonging to the third-generation caused by the development of resistance. In this review, we have summarized the work done in the last five years to overcome the limitations of currently marketed drugs, giving structural activity relationships of quinazoline-based lead compounds synthesized and tested recently. We have also highlighted the shortcomings of the currently used approaches and have provided guidance for circumventing these limitations. Our review would help medicinal chemists streamline and guide their efforts towards developing novel quinazoline-based EGFR inhibitors.


The influence of the systematic birth preparation program on childbirth satisfaction.
The primary purpose of this study was to assess the influence of a systematic multidisciplinary birth preparation program on satisfaction with childbirth experience. A secondary aim was to detect factors that affect the childbirth satisfaction. In this prospective study, 77 pregnant women who completed the 4-month birth preparation program (Group 1) and 75 women in the control group (Group 2) were asked to fill out two questionnaires with face-to-face interviews within 48 h after labor. One of the questionnaires was the translated form of Salmon's Item List German (SIL-Ger), and SIL-Ger scores ≥70 was accepted as a satisfactory experience. There was no statistically significant difference between the groups in terms of sociocultural and intrapartum characteristics, and obstetric outcome parameters. The women who received antenatal education experienced significantly less pain (p = 0.01), had a better communication with midwife or obstetrician during delivery (p = 0.001), and participated more actively in decision-making before, during, and after childbirth (p < 0.001 for all). SIL score was significantly higher in Group 1 (105.7 ± 2.2 vs 80 ± 2.5, p < 0.01), and significantly more women had a SIL score ≥70 (96.1 vs 73.3%, p < 0.01). In multivariate logistic regression model, attending the birth preparation program and the level of pain perceived during labor were found to have a significant effect on the birth satisfaction. Systematic birth preparation program improves satisfaction with childbirth experience by enabling women to communicate better with healthcare providers and to participate in decision-making during labor, as well as by decreasing the perception of labor pain.


CNT-molecule-CNT (1D-0D-1D) van der Waals integration ferroelectric memory with 1-nm2 junction area.
The device's integration of molecular electronics is limited regarding the large-scale fabrication of gap electrodes on a molecular scale. The van der Waals integration (vdWI) of a vertically aligned molecular layer (0D) with 2D or 3D electrodes indicates the possibility of device's integration; however, the active junction area of 0D-2D and 0D-3D vdWIs remains at a microscale size. Here, we introduce the robust fabrication of a vertical 1D-0D-1D vdWI device with the ultra-small junction area of 1 nm2 achieved by cross-stacking top carbon nanotubes (CNTs) on molecularly assembled bottom CNTs. 1D-0D-1D vdWI memories are demonstrated through ferroelectric switching of azobenzene molecules owing to the cis-trans transformation combined with the permanent dipole moment of the end-tail -CF3 group. In this work, our 1D-0D-1D vdWI memory exhibits a retention performance above 2000 s, over 300 cycles with an on/off ratio of approximately 105 and record current density (3.4 × 108 A/cm2), which is 100 times higher than previous study through the smallest junction area achieved in a vdWI. The simple stacking of aligned CNTs (4 × 4) allows integration of memory arrays (16 junctions) with high device operational yield (100%), offering integration guidelines for future molecular electronics.


The effects of Triton WR-1339, protamine sulfate and heparin on the plasma removal of emulsion models of chylomicrons and remnants in rats.
Protein-free lipid emulsions with compositions modelling chylomicrons (chylomicron-like emulsion) or chylomicron remnants (remnant-like emulsion) were injected intra-arterially into nonanesthetized rats. Compared with control untreated rats, treatment with Triton WR-1339, protamine sulfate or heparin strongly modified the plasma removal of triacylglycerols and cholesteryl ester moieties of chylomicron-like emulsions, but had little effect on removal rates of triacylglycerols or cholesteryl esters of remnant-like emulsions. The effects on chylomicron-like removal were similar to those on natural lymph chylomicrons. The relative lack of effects on remnant-like emulsion removal provides additional evidence that remnant-like emulsions are a metabolic model for natural chylomicron remnants.


Contrastive Learning for Prediction of Alzheimer's Disease Using Brain 18F-FDG PET.
Brain 18F-FDG PET images are commonly-known materials for effectively predicting Alzheimer's disease (AD). How-ever, the data volume of PET is usually insufficient, which is unfavorable to train an accurate AD prediction networks. Fur-thermore, the PET image is noisy with low signal-to-noise ratio, and simultaneously the feature (metabolic abnormality) used for predicting AD in PET image is not always obvious. Such charac-teristics of 18F-FDG PET images hinder the existing deep learning networks to learn the feature of lesion (i.e., glucose metabolism abnormality) effectively, which leads to unsatisfactory classifica-tion performance and poor robustness. In this paper, a contrastive-based learning method is proposed to address the challenges of PET image inherently possessed. Firstly, the slices of 3D PET image are amplified by cropping the image of anchors (i.e., an augmented version of the same image) to generate extended train-ing data. Meanwhile, contrastive loss is adopted to enlarge inter-class feature distances and reduce intra-class feature differences using subject fuzzy labels as supervised information. Secondly, we construct a double convolutional hybrid attention module to enhance the network to learn different perceptual domains where two convolutional layers with different convolutional kernels (7 × 7 and 5 × 5) are constructed. Moreover, we recommend a diagnosis mechanism by analyzing the consistency of predicted result for PET slices alone with clinical neuropsychological assessment to achieve a better AD diagnosis. The experimental results show that the proposed method outperforms the state-of-the-arts for brain 18F-FDG PET images while remaining satisfactory computational performance, and hence demonstrate the advantage of the method in effectively predicting AD.


Intravitreal ranibizumab for pigment epithelium detachment with subfoveal occult choroidal neovascularization: a prospective 24-month case series.
To assess the effects of intravitreal ranibizumab injection in patients affected by pigment epithelial detachment associated with occult subfoveal choroidal neovascularization. Prospective, interventional case series. Forty eyes of 40 patients were considered for the purpose of the study. Consecutive patients were recruited for a 24-month study. All patients underwent a complete ophthalmic examination, including best-corrected visual acuity on Early Treatment Diabetic Retinopathy Study (ETDRS) charts. After a 3-monthly loading phase, further intravitreal ranibizumab injections were administered on the basis of detection of any type of fluid on optical coherence tomography. Changes in mean best-corrected visual acuity at 12 and 24 months and the proportion of eyes losing fewer than 15 letters (corresponding to 3 ETDRS lines) from baseline visual acuity. Changes in central macular thickness on optical coherence tomography and variation in mean area of the entire lesion. Forty patients were included. Mean best-corrected visual acuity decreased from 20/66 (58 ETDRS letters) to 20/83 (53 letters) at 12 months and 20/112 (489 ETDRS letters) at 24 months (P = .003). Eighty percent and 67.5% of eyes lost fewer than 3 lines at 12 and 24 months, respectively. Mean central macular thickness passed from 545 μm to 428 μm at 12 months and 426 μm at 24 months. Mean lesion area changed from 6826 μm(2) to 6312 μm(2) at 12 months and 6010 μm(2) at 24 months. The treatment of pigment epithelial detachment associated with occult subfoveal choroidal neovascularization with intravitreal ranibizumab injection after a 3-monthly loading phase and pro re nata strategy can lead to partial results over a 24-month follow-up. Further investigations are warranted to establish the best therapeutic approach to this disease.


Cell-Free Synthesis of Plant Receptor Kinases.
The wheat germ cell-free protein synthesis system has been used as a eukaryotic protein production system since it was first reported in 1964. Although initially the productivity of this system was not very high, it has now become one of the most versatile protein production systems, thanks to the enhancements made by several groups. In this chapter, we report a protein production method for plant receptor kinases using the wheat cell-free system. We describe a method for the preparation of a cell-free extract from wheat germ, the split-primer PCR method for preparation of transcription templates, and the bilayer cell-free protein synthesis method.


Antiretroviral therapy and HIV-associated cardiovascular disease: a prospective cardiac biomarker and CMR tissue characterization study.
Biochemical markers are fundamental in cardiac evaluation, and various novel assays have recently been discovered. We prospectively evaluated the hearts of newly diagnosed people living with human immunodeficiency virus (PLWH) using cardiac biomarkers, compared them with human immunodeficiency virus (HIV)-uninfected controls, and correlated our prospective findings with cardiovascular magnetic resonance imaging (CMR). Newly diagnosed, antiretroviral therapy (ART)-naïve PLWH were recruited along with HIV-uninfected, age-matched, and sex-matched controls. All participants underwent measurement of high-sensitivity cardiac troponin T (hs-cTnT), N-terminal pro-B-type natriuretic peptide (NT-proBNP), soluble ST2 (sST2), and galectin-3, as well as a CMR study with multiparametric mapping. The HIV group started ART and was re-evaluated 9 months later. The cardiac biomarkers and their correlation with CMR parameters were evaluated in and between groups. Compared with controls (n = 22), hs-cTnT (4.0 vs. 5.1 ng/L; P = 0.004), NT-proBNP (23.2 vs. 40.8 ng/L; P = 0.02), and galectin-3 (6.8 vs. 9.0 ng/mL; P = 0.002) were all significantly higher in the ART-naïve group (n = 73). After 9 months of ART, hs-cTnT (5.1 vs. 4.3 ng/L; P = 0.02) and NT-proBNP (40.8 vs. 28.5 ng/L; P = 0.03) both decreased significantly and a trend of decrease was seen in sST2 (16.5 vs. 14.8 ng/L; P = 0.08). Galectin-3 did not demonstrate decrease over time (9.0 vs. 8.8 ng/mL; P = 0.6). The cardiac biomarkers that showed the best correlation with CMR measurements native T1, T2, and extracellular volume were NT-proBNP (rs ≥ 0.4, P < 0.001) and galectin-3 (rs ≥ 0.3, P < 0.01). Our cardiac biomarker data support the presence of subclinical myocardial injury, remodelling, and fibrosis at HIV diagnosis, and ART had a positive influence on these blood markers. It remains unclear if the underlying pathological processes were fully addressed by ART. The ability of cardiac biomarkers to detect and track tissue abnormalities diagnosed with CMR showed promise. With additional research, this could lead to improvements in screening and monitoring myocardial abnormalities, even in CMR-limited settings.


GINA 2020: Potential Impacts, Opportunities, and Challenges for Primary Care.
In 2019, it was reported that changes to asthma management reported in the Global Initiative for Asthma (GINA) "…might be considered the most fundamental changes in asthma management in 30 years." These changes refer to the recommendation that the treatment of asthma in adolescents and adults would no longer include short-acting ß2-agonist (SABA) only, but that people with asthma should receive either symptom-driven inhaled corticosteroids (ICS)-containing treatment (mild asthma) or daily ICS-containing treatment. The fundamental reason for this shift was driven by concerns about the risks and consequences associated with SABA-only treatment, the need to improve the day-to-day management of asthma symptoms to prevent exacerbations and emergent evidence. These recommendations have subsequently been reinforced and characterized in GINA 2020, and it is reasonable to say that they are significant, not only in terms of an asthma management framework but also as a management approach in practice. This opinion article specifically focuses on opportunities and challenges associated with the implementation of GINA 2020 in primary care practice that need to be recognized and addressed if the shift in asthma treatment paradigm is to be successfully implemented into day-to-day practice.


Functional characterization of the CDF transporter SMc02724 (SmYiiP) in Sinorhizobium meliloti: Roles in manganese homeostasis and nodulation.
In bacteria, membrane transporters of the cation diffusion facilitator (CDF) family participate in Zn(2+), Fe(2+), Mn(2+), Co(2+) and Ni(2+) homeostasis. The functional role during infection processes for several members has been shown to be linked to the specificity of transport. Sinorhizobium meliloti has two homologous CDF genes with unknown transport specificity. Here we evaluate the role played by the CDF SMc02724 (SmYiiP). The deletion mutant strain of SmYiiP (ΔsmyiiP) showed reduced in vitro growth fitness only in the presence of Mn(2+). Incubation of ΔsmyiiP and WT cells with sub-lethal Mn(2+) concentrations resulted in a 2-fold increase of the metal only in the mutant strain. Normal levels of resistance to Mn(2+) were attained by complementation with the gene SMc02724 under regulation of its endogenous promoter. In vitro, liposomes with incorporated heterologously expressed pure protein accumulated several transition metals. However, only the transport rate of Mn(2+) was increased by imposing a transmembrane H(+) gradient. Nodulation assays in alfalfa plants showed that the strain ΔsmyiiP induced a lower number of nodules than in plants infected with the WT strain. Our results indicate that Mn(2+) homeostasis in S. meliloti is required for full infection capacity, or nodule function, and that the specificity of transport in vivo of SmYiiP is narrowed down to Mn(2+) by a mechanism involving the proton motive force.


Implication of expression of Nanog in prostate cancer cells and their stem cells.
Recent studies suggested that the prostate cancer may arise from prostate cancer stem cells that share some same characteristics with normal stem cells. The purpose of this study was to detect the differences of Nanog expression between PC3 prostate cancer cell line and its tumor stem cells, and the relationship was preliminarily examined between Nanog and prostate cancer and its tumor stem cells. By using magnetic active cell sorting (MACS), we isolated a population of CD44(+)/CD133(+) prostate cancer cells that display stem cell characteristics from PC3 cell line. Immunohistochemistry revealed positive expressions of CD44, CD133 and α(2)β(1)-integin in the isolated cells. CCK-8 analysis showed that isolated cells had a strong proliferative ability. The formation of the cell spheres in serum-free medium and holoclones in serum-supplied medium showed that the cells were capable of self-renewing, indicating that the isolated cells were a population of cancer stem-like cells derived from PC3 cell line. Western blotting exhibited that the isolated cells had higher experession of Nanog, an embryonic stem marker, as compared with PC3 cells. Our study showed that Nanog might be helpful in sustaining the self-renewal and the undifferentiation of prostate cancer stem cells, and may serve as a marker for prostate cancer stem cells for isolation and identification.


A molecular phylogeny of Acronychia, Euodia, Melicope and relatives (Rutaceae) reveals polyphyletic genera and key innovations for species richness.
We present the first detailed phylogenetic study of the genus Melicope, the largest genus of the Citrus family (Rutaceae). The phylogenetic analysis sampled about 50% of the 235 accepted species of Melicope as well as representatives of 26 related genera, most notably Acronychia and Euodia. The results based on five plastid and nuclear markers have revealed that Acronychia, Euodia and Melicope are each not monophyletic in their current circumscriptions and that several small genera mainly from Australia and New Caledonia need to be merged with one of the three genera to ensure monophyly at the generic level. The phylogenetic position of the drupaceous Acronychia in relation to Melicope, which has capsular or follicular fruits, remains unclear and Acronychia might be a separate genus or a part of Melicope. The seed coats of Melicope, Acronychia and related genera show adaptations to bird-dispersal, which might be regarded as key innovations for species radiations. Euodia and its relatives, which lack these adaptations, include only about 20 species while the Melicope-Acronychia group consists of about 340 species. The drupaceous genera Comptonella, Dutaillyea, Picrella and Sarcomelicope are nested within Melicope and need to be merged with Melicope. The expanded genus is a prime example of the artificial classification system of Engler, who defined Rutaceous subfamilies mainly based on gynoecial and fruit characters.


CT, MRI, and color Doppler ultrasound correlation of pancreatoblastoma: a case report.
Pancreatic neoplasms in childhood are rare but should be considered in the differential diagnosis of masses in the region of the lesser sac. A case of pancreatoblastoma in a 3 1/2-year-old boy is presented. The patient was studied with US, CT, and MR imaging.


The association between lithium use and neurocognitive performance in patients with bipolar disorder.
Lithium remains the gold standard for the treatment of bipolar disorder (BD); however, its use has declined over the years mainly due to the side effects and the subjective experience of cognitive numbness reported by patients. In the present study, we aim to methodically test the effects of lithium on neurocognitive functioning in the largest single cohort (n = 262) of BD patients reported to date by harnessing the power of a multi-site, ongoing clinical trial of lithium monotherapy. At the cross-sectional level, multivariate analysis of covariance (MANCOVA) was conducted to examine potential group differences across neurocognitive tests [California Verbal Learning Test (CVLT trials 1-5,CVLT delayed recall), Wechsler Digit Symbol, Trail-making Test parts A and B (TMT-A; TMT-B), and a global cognition index]. At the longitudinal level, on a subset of patients (n = 88) who achieved mood stabilization with lithium monotherapy, we explored the effect of lithium treatment across time on neurocognitive functioning. There were no differences at baseline between BD patients that were taking lithium compared with those that were not. At follow-up a significant neurocognitive improvement in the global cognitive index score [F = 31.69; p < 0.001], CVLT trials 1-5 [F = 29.81; p < 0.001], CVLT delayed recall [F = 15.27; p < 0.001], and TMT-B [F = 6.64, p = 0.012] was detected. The cross-sectional and longitudinal (on a subset of 88 patients) investigations suggest that lithium may be beneficial to neurocognitive functioning in patients with BD and that at the very least it does not seem to significantly impair cognition when used therapeutically.


Detection of Tumor Recurrence via Circulating Tumor DNA Profiling in Patients with Localized Lung Cancer: Clinical Considerations and Challenges.
Approximately 30% of patients with non-small-cell lung cancer (NSCLC) present with localized/non-metastatic disease and are eligible for surgical resection or other "treatment with curative intent". Due to the high prevalence of recurrence after treatment, adjuvant therapy is standard care for most patients. The effect of adjuvant chemotherapy is, however, modest, and new tools are needed to identify candidates for adjuvant treatments (chemotherapy, immunotherapy, or targeted therapies), especially since expanded lung cancer screening programs will increase the rate of patients detected with localized NSCLC. Circulating tumor DNA (ctDNA) has shown strong potential to detect minimal residual disease (MRD) and to guide adjuvant therapies. In this manuscript, we review the technical aspects and performances of the main ctDNA sequencing platforms (TRACERx, CAPP-seq) investigated in this purpose, and discuss the potential of this approach to guide or spare adjuvant therapies after definitive treatment of NSCLC.


Perioperative Outcomes After Preoperative Epidural Analgesia in Patients with Hip Fracture Undergoing Surgical Repair: A Systematic Review.
To examine the effectiveness and safety of epidural analgesia in the presurgical period in patients with hip fracture undergoing surgical repair. Systematic review. The study protocol was registered with the PROSPERO systematic reviews registry with the (identifier CRD42019140396). Electronic databases were searched for randomized controlled trials comparing preoperative epidural analgesia with other forms of pain management in patients with a hip fracture. The primary outcomes included perioperative cardiac events and death. Pain, noncardiac complications, and adverse effects were also examined as secondary outcomes. The heterogeneity of the included studies was assessed with the I2 statistic, and a random-effects meta-analysis was conducted once sufficient homogeneity was demonstrated. Four studies, which included a total of 221 patients, met the inclusion criteria. Preoperative epidural analgesia resulted in fewer cardiac events, which was a reported outcome in two included studies (relative risk 0.30; 95% confidence interval 0.14-0.63; I2=0%). Preoperative epidural analgesia was also associated with a decreased perioperative mortality rate in a meta-analysis of two studies (relative risk 0.13; 95% confidence interval 0.02-0.98; I2 = 0%). Pain was not pooled because of variability in assessment methods, but preoperative epidural analgesia was associated with reduced pain in all four studies. Preoperative epidural analgesia for hip fracture may reduce perioperative cardiac events and deaths, but the number of included studies in this systematic review was low. More research should be done to determine the benefit of early epidural analgesia for patients with hip fracture.


Membrane mobility agent alters the consequences of lectin-cell interaction in a malignant cell membrane.
The membrane mobility agent 2-(2-methoxyethoxy)-ethyl 8-cis-2-n-octyl-cyclopropyl)-octanoate promotes cap formation from wheat germ agglutinin-receptor combinations at the expense of agglutination in membranes of malignant mastocytoma cells.


Current topic: the placental corticotrophin-releasing hormone-adrenocorticotrophin axis.
In this review the factors regulating production of corticotrophin-releasing hormone (CRH) in intrauterine tissues are discussed and interactions of placental CRH with placental pro-opiomelanocortin (POMC)/adrenocorticotrophin (ACTH) and prostaglandins (PG) are examined. Discrepant results concerning localization of immunoreactive (IR-) CRH and its binding protein (CRH-BP) and their mRNAs in intrauterine tissues are described, and these issues require further resolution. It is suggested that the CRH-ACTH-PG axis in the placenta and choriodecidua may be important in relation to paracrine/autocrine regulation of uteroplacental blood flow, and in term and preterm labour. During the initial stages of preterm labour in the setting of infection, intrauterine cytokines and other factors may stimulate CRH output, implying a role for the immune-neuroendocrine axes in this process. With loss of chronic trophoblasts in advanced infection leading to preterm labour, a major intrauterine site of CRH production may be lost and the influence of this pathway becomes minimal. At this time increased intrauterine prostaglandin synthesis, together with loss of prostaglandin dehydrogenase activity in the fetal membranes, may become the primary route leading to myometrial activity and delivery.


Adjuvant effects of bacterium-like particles in the intranasal vaccination of chickens against Newcastle disease.
Given that the respiratory mucosa is an important site for the initial replication of Newcastle disease virus (NDV), developing intranasal vaccines for chickens is an effective strategy to protect against this disease. The low immunogenicity of inactivated NDV administered by the mucosal route motivated us to identify a safe and potent adjuvant. Previous studies have shown that bacterium-like particles (BLPs), which serve as mucosal adjuvants, induce effective local and systemic immune responses through TLR2 signaling in both mammals and humans. Here, we report that BLPs could activate the innate immune system of chickens in a manner that was dependent on the combination of chicken TLR2 type 1 (chTLR2t1) and chicken TLR1 type 1 (chTLR1t1). The chicken macrophage-like HD11 cell line was stimulated with BLPs, resulting in the production of nitric oxide and the expression of the proinflammatory cytokines IFN-γ, IL-1β and IL-6. Chickens intranasally immunized with inactivated NDV vaccines mixed with BLP adjuvants exhibited significantly increased levels of local SIgA in their tracheal lavage fluid and as well as hemagglutination-inhibiting antibodies in serum samples. The strong systemic and local immune responses induced by BLP-adjuvanted vaccines provided 100 % protection against intranasal challenge with a lethal dose of virulent NDV without showing any signs of disease. These results indicate that BLPs should be considered for use as a potential mucosal adjuvant for inactivated NDV vaccines and other vaccines for poultry.


Generalized joint hypermobility and risk of pelvic girdle pain in pregnancy: does body mass index matter?
Women with generalized joint hypermobility may be at increased risk of pregnancy-related pelvic girdle pain, but evidence is inconclusive. In this prospective cohort study of 283 pregnant women in Norway, we aimed to study the association of generalized joint mobility with pelvic girdle pain, and to evaluate if pre-pregnancy body mass index was a modifier of the association. Generalized joint hypermobility was defined as a score of ≥5/9 positive tests on the Beighton score measured in early pregnancy. Primary outcome was evening pain intensity in gestation week 30, measured by a 100 mm visual analogue scale. We applied linear regression analyses to estimate age-adjusted unstandardized beta coefficients. Evening pain intensity was similar among women with Beighton score ≥ 5/9 and women with Beighton score < 5/9 (age-adjusted mean difference 2.8 mm; 95% CI: -9.2 to 14.9 mm). Women with Beighton score ≥ 5/9 and pre-pregnancy body mass index ≥ 25 kg/m2, reported higher evening pain than women with Beighton score < 5/9 and pre-pregnancy body mass index <25 kg/m2 (age-adjusted mean difference 28.7 mm; 95% CI: 14.3-43.1 mm). Overall, evening pain intensity was similar among pregnant women with and without generalized joint hypermobility. However, women with a combination of generalized joint hypermobility and body mass index ≥25 kg/m2 reported higher evening pain compared to women with normal joint mobility and body mass index <25 kg/m2, suggesting that body mass index may modify the association. The estimates could be imprecise due to the small study sample, and our findings should be interpreted with caution.


A Novel Combined DNA Vaccine Encoding Toxoplasma gondii SAG1 and ROP18 Provokes Protective Immunity Against a Lethal Challenge in Mice.
Antigens expressed by Toxoplasma gondii (T. gondii) during its life cycle trigger various immune responses in the host. Recently, toxoplasma vaccine research focused on T. gondii surface antigen 1 (SAG1) and Rhoptry Protein 18 (ROP18) to establish a safe and efficacious DNA vaccine. We constructed two eukaryotic expression plasmids: p3 × FLAG-Myc-CMV™-24-SAG1 and p3 × FLAG-Myc-CMV™-24-ROP18. BALB/c mice were randomly divided into six groups and immunized with these DNA vaccines either separately or in combination. The combination vaccine was administered at either the full dose or at half-strength dose. Control mice were immunized with empty vector or with phosphate-buffered saline. The frequency of CD4+ cells in the spleen was consistent among all groups, whereas that of CD8+ T cells was the highest in the group immunized with the combination vaccine at half-strength dose (p < 0.05). Importantly, the mRNA expression levels of interleukin-12 (IL-12) and interferon-gamma (INF-γ) were closely correlated (r = 0.6, p < 0.0001) and both were upregulated in the group that was immunized with the combination vaccine at half-strength dose (p < 0.0001). The survival time of the mice subjected to a lethal dose of toxoplasma was significantly extended by prior immunization with DNA vaccines expressing either SAG1 or ROP18 or a combination of both (p < 0.05). The group that was immunized with the combination vaccine at half-strength dose demonstrated the best efficacy (p < 0.05). These results showed that the combination DNA vaccine provided better immune protection than the single gene vaccines, and that optimizing the dosing of the vaccine can improve the immune response.


Molecular insights into the germline for prostate cancer initiation, progression, and aggressiveness.
Germline and tumor genetic testing of DNA repair genes in men with advanced prostate is increasingly recommended by U.S. and international guidelines as part of standard of care. Damaging mutations in homologous DNA repair pathways genes including BRCA2, BRCA1, PALB2, and ATM, and mismatch DNA repair genes including MSH2 and MSH6 have emerging clinical utility for risk assessment and treatment decision-making. This article summarizes a presentation at the 2019 Philadelphia Consensus Conference focused on the latest data at the intersection of germline and tumor genetic testing for prostate cancer patients.


Relationships between cognition and structural neuroimaging findings in adults with human immunodeficiency virus type-1.
Infection of the central nervous system with human immunodeficiency virus type-1 (HIV) is associated with cognitive impairments that range from mild cognitive and motor difficulties to dementia. Structural neuroimaging abnormalities are also common in HIV-infected patients both with and without cognitive disturbances. The most common abnormalities include high signal intensities in the white matter and atrophy. Research over the past 12 years has helped define the relationship between these neuroimaging abnormalities and the manifestation of cognitive disturbance in HIV. In the present paper, we provide a synopsis of these studies and report the current state of the literature. Our review revealed that atrophy of the caudate nucleus is most consistently associated with cognitive impairment in HIV. The current literature does not support a strong relationship between cortical atrophy or white matter abnormalities and cognitive dysfunction in this population, though methodological issues may have influenced the results. Suggestions for study design and new research directions are provided.


Roles of ribosomal proteins in the functioning of translational machinery of eukaryotes.
Ribosomal proteins (rps) are constituents of ribosomal subunits together with the rRNAs, and the work of the ribosomes is based on highly cooperative interactions involving these biopolymers. Eukaryotic ribosomal subunits contain proteins that have bacterial counterparts as well as proteins specific only to eukaryotes and archaea and proteins unique to eukaryotes. Roles of eukaryotic rps in functioning of the translation machinery are studied in less detail than those of bacterial ones. However, application of various structural, biochemical and genetic approaches made it possible to obtain data suggesting implication of many particular eukaryotic rps in binding of participants of translation process (ribosomal ligands). Here we present a review on these data as well as their analysis aimed to reveal conserved and eukaryote-specific functions of eukaryotic rps in the work of translational machinery.


Ferrocene conjugated oligonucleotide for electrochemical detection of DNA base mismatch.
We describe the synthesis, binding, and electrochemical properties of ferrocene-conjugated oligonucleotides (Fc-oligos). The key step for the preparation of Fc-oligos contains the coupling of vinylferrocene to 5-iododeoxyuridine via Heck reaction. The Fc-conjugated deoxyuridine phosphoramidite was used in the Fc-oligonucleotide synthesis. We show that thiol-modified Fc-oligos deposited onto gold electrodes possess potential ability in electrochemical detection of DNA base mismatch.


Formation of RACK- and grid-type metallosupramolecular architectures and generation of molecular motion by reversible uncoiling of helical ligand strands.
The interaction of appropriate metal ions (Pb(II), Zn(II)) with helical ligand strands, obtained by hydrazone polycondensation, generates polymetallic supramolecular architectures of rack and grid types, by uncoiling of the ligand. The interconversion between the helical free ligand and the linearly extended ligand in the complexes produces reversible ion-induced, nanomechanical molecular motions of large amplitude. It has been integrated in an acid-base neutralisation fuelled process, which links the extension/contraction of the ligand strands to alternating changes in pH.


Postmortem changes in uric acid and ascorbic acid in human cerebral cortex tissues excised after cardiac death.
There has been no report on the determination of uric acid (UA) in human brain and heart tissues. UA and ascorbic acid (AA) in human cerebral cortex and heart tissues excised after cardiac death have been studied by reversed-phase high-performance liquid chromatography (HPLC) with electrochemical detection (ECD). It has been found that the levels of AA and UA in the human cerebral cortex tissues tend to decrease and increase, respectively, after cardiac death as a function of time between death and forensic operation. In addition, it has been found that there is no special relationship between UA levels in human heart tissues and time after cardiac death, also that the UA levels in the heart are high as compared with those in human cerebral cortex tissues. We have emphasized that the HPLC-ECD method is useful in determining UA and AA in mammalian tissues by one-time chromatography to gain a better understanding of the relationship between disease and serum urate level.


A spectroscopic approach to measuring meibum lipid composition and conformation in donors with Sjӧgren's syndrome.
Patients with Sjӧgren's syndrome (SS) have dry eye associated with meibomian gland dysfunction (MGD). The meibum from donors with dry eye due to MGD but without SS (MMGD) presents with lower levels of cholesteryl ester, less straight chains, and more ordered hydrocarbon chains compared with meibum from donors without MGD (Mn). The aim of the current study was to compare the composition and hydrocarbon chain conformation of meibum from donors with Sjögren's syndrome (Mss) to Mn and MMGD. Meibum was expressed from patients with SS using an ILUX instrument (Alcon Inc., Fort Worth TX). All of the nine meibum donors with SS were female. Meibum composition was characterized using 1H-NMR and meibum hydrocarbon chain conformation was measured using fourier transform infrared spectroscopy. Meibum from every donor with SS measured contained a significantly (P < 0.01) higher cholesteryl ester/wax ester ratio and more straight chains compared with donors without SS or dry eye. None of the nine phase transitional parameters were significantly different, P > 0.05, for Mss compared with Mn. Nor was the CH3/CH2 band height ratio used to estimate the number of hydrocarbon CH3 and CH2 moieties different, P = 0.22, for Mss compared with Mn. In conclusion, the compositional differences between Mss compared with Mn did not result in differences in any of the nine meibum lipid phase transitional parameters measured. The compositional differences observed between Mss and Mn could be markers for or contribute to SS as the differences could lead to tear film lipid packing differences other than conformational differences.


A case of left-sided absence and right-sided fenestration of the external jugular vein and a review of the literature.
The external jugular vein is increasingly being used as the recipient vein in head and neck tissue transfers, and for cannulation to conduct diagnostic procedures or intravenous therapies. The variations in the patterns of its course, and knowledge of these variations, are therefore important. We report on a bilateral external jugular vein anomaly found during the neck dissection of an approximately 75-year-old female cadaver, a case which has hitherto not been reported. The vein was absent on the left and fenestrated on the right. An embryological evaluation and the clinical implications of the anomaly are described. Clinicians and surgeons performing vascular or reconstructive surgery should be made aware of this variation of the external jugular vein to prevent inadvertent injury.


Red ginseng delays age-related hearing and vestibular dysfunction in C57BL/6 mice.
Since Korean red ginseng (KRG) has been proven to protect against gentamicin-induced vestibular and hearing dysfunction, the effects of KRG on age-related inner ear disorder in C57BL/6 mice were investigated. While age-related hearing loss was detected at the age of 6months (32kHz) and 9months (16kHz) in the control group, it was significantly delayed (p<0.05) in the 150mg/kg KRG-treated group. Vestibular dysfunction was observed in the tail-hanging and swimming tests, with significantly different severity scores and swimming times detected between the control and 150mg/kg KRG-treated group at the age of 12months (p<0.05). Mice treated with 500mg/kg KRG exhibited irritability and aggravated inner ear dysfunction. Histological observation supported the findings of hearing and vestibular function defects. In conclusion, C57BL/6 mice showed early-onset hearing loss and progressive vestibular dysfunction with aging, which were delayed by treatment with 150mg/kg KRG. However, 500mg/kg KRG treatment may induce aggressive behavior.


Delayed onset and decreased incidence of diabetes in BB rats fed free radical scavengers.
We tested the hypothesis that free radicals play a role in the selective destruction of pancreatic beta-cells in BB/Wor rats. Diabetes-prone BB rats of both sexes and 40 days of age were divided into three groups. The control group was fed ad libitum Purina rat chow powder, while the experimental group was fed ad libitum the rat chow powder blended with a mixture of four known free radical scavengers: allopurinol, mercaptopropionylglycine, dimethylthiourea and Vitamin E. A third group was pair-fed 10 g chow powder/rat/day, since in earlier experiments we observed that rats on the experimental diet consumed only about 10 g/rat/day. All rats were studied up to age 120 days. Body weight and food intake were measured daily. Urine was tested for glucose beginning at age 60 days. When glucosuria appeared, blood glucose and urinary ketones were measured. Body weight gain in the experimental and pair-fed groups was similar, but lower than the control group. Life table analysis of the data showed a decreased and a delayed onset of diabetes in the rats fed free radical scavengers. Thus, the results of this study demonstrated that calorie restriction and the related impaired growth did not affect the incidence of diabetes in the BB rat. In addition, the results suggested a role for free radicals in the spontaneous destruction of pancreatic beta-cells in the BB rat.


Spray-dried didanosine-loaded polymeric particles for enhanced oral bioavailability.
Didanosine (ddI) is a water-soluble antiretroviral used in the treatment of HIV that undergoes fast gastric degradation to an inactive hypoxanthine. Therefore, its oral bioavailability is relatively low (20-40%). In this work, we investigated for the first time a scalable open-loop spray-drying method with co-current flow for the encapsulation of ddI (model drug) within particles of the biocompatible polyester poly(epsilon-caprolactone). The average diameter of the particles was 36-118 μm and the morphology spherical. The encapsulation efficiency ranged from 60% to 100% with yields of up to 65%. ATR/FT-IR analysis indicated that most of the drug was encapsulated within the particles. In vitro release assays showed that the particles released the drug within 120 min. Finally, oral administration to rats led to a statistically significant 2.5-fold increase of the bioavailability with respect to a ddI aqueous solution, highlighting the potential of this technology to encapsulate efficiently other hydrophilic antiretrovirals and, by doing so, to overcome different biopharmaceutical drawbacks associated with the oral administration.


Assessing the habitat suitability of agricultural landscapes for characteristic breeding bird guilds using landscape metrics.
Many of the processes behind the decline of farmland birds can be related to modifications in landscape structure (composition and configuration), which can partly be expressed quantitatively with measurable or computable indices, i.e. landscape metrics. This paper aims to identify statistical relationships between the occurrence of birds and the landscape structure. We present a method that combines two comprehensive procedures: the "landscape-centred approach" and "guild classification". Our study is based on more than 20,000 individual bird observations based on a 4-year bird monitoring approach in a typical agricultural area in the north-eastern German lowlands. Five characteristic bird guilds, each with three characteristic species, are defined for the typical habitat types of that area: farmland, grassland, hedgerow, forest and settlement. The suitability of each sample plot for each guild is indicated by the level of persistence (LOP) of occurrence of three respective species. Thus, the sample plots can be classified as "preferred" or "less preferred" depending on the lower and upper quartiles of the LOP values. The landscape structure is characterized by 16 different landscape metrics expressing various aspects of landscape composition and configuration. For each guild, the three landscape metrics with the strongest rank correlation with the LOP values and that are not mutually dependent were identified. For four of the bird guilds, the classification success was better than 80%, compared with only 66% for the grassland bird guild. A subset of six landscape metrics proved to be the most meaningful and sufficiently classified the sample areas with respect to bird guild suitability. In addition, derived logistic functions allowed the production of guild-specific habitat suitability maps for the whole landscape. The analytical results show that the proposed approach is appropriate to assess the habitat suitability of agricultural landscapes for characteristic bird guilds.


Is varus-valgus constraint a reliable option in complex primary total knee arthroplasty? A systematic review.
Knee instability is considered one of the most frequent cause of failure after primary total knee arthroplasty (TKA). In order to address intraoperative instability, varus-valgus constrained knee implants (VVC) are increasingly utilized in primary TKA. Despite an increased risk of mechanical failure, short to mid-term results seem to be encouraging, but long-term results are still lacking. A systematic review of prospective and retrospective studies that reported clinical outcomes of patients with VVC systems in primary TKAs between 1990 and 2020 was performed. In all, 28 articles met our inclusion criteria. A total of 2798 VVC implants were used in primary TKA. The all-cause revision-free survivorship was 95.2% at a mean follow-up of 7 years. Infection and aseptic loosening were the most common reasons for reoperation with an incidence of 1.8% and 1.7%, respectively. Overall complication rate was 9.6%, the most common complications were knee stiffness and infection with an incidence of 2.8% and 2.5%, respectively. VVC implants in primary TKA are associated with improved functional outcomes and good mid-term survivorship, comparable to lower level of constraint implants. Non-modular stemless seem to be reliable implants at mid-term follow-up. However, given the lack data coming from long-term studies, VVC implants should be used cautiously in primary TKA.


Minor neurological dysfunction is more closely related to learning difficulties than to behavioral problems.
In a group of 570 nine-year-old children (315 boys and 255 girls) without an overtly handicapping neurological condition, relationships were studied between the presence of minor neurological dysfunction (MND) on the one hand and cognitive and behavioral problems on the other hand. The aim of the study was to investigate whether MND was more closely related to learning problems than to behavioral difficulties. The group was a subpopulation of the birth cohort of the Groningen Perinatal Project in the Netherlands, in which perinatal developmental relationships are studied. The age-specific and standardized neurological examination technique of Touwen (1979) was used, resulting in a description of the neurological condition in terms of the number of deviant clusters. The absence of deviant clusters indicated a normal neurological condition (n = 418); children with one or two deviant clusters were classified as MND-1 (n = 86) and those with more than two deviant clusters as MND-2 (n = 66). The presence and severity of MND was significantly related to poor performance on standardized reading, spelling, and arithmetic tests. MND was also related to ratings of distractible behavior on parent and teacher questionnaires, but to a lesser extent than the cognitive problems. Ratings of so-called "troublesome" and "timid" behavior were not related to MND. Our conclusion is that learning problems are more closely related to MND than are behavioral difficulties. This has implications for prevention and intervention: In the former the focus should be more on biological hazards, in the latter on environment and rearing attitudes.


Combination of Talazoparib and Calcitriol Enhanced Anticancer Effect in Triple-Negative Breast Cancer Cell Lines.
Monotherapy for triple-negative breast cancer (TNBC) is often ineffective. This study aimed to investigate the effect of calcitriol and talazoparib combination on cell proliferation, migration, apoptosis and cell cycle in TNBC cell lines. Monotherapies and their combination were studied for (i.) antiproliferative effect (using real-time cell analyzer assay), (ii.) cell migration (CIM-Plate assay), and (iii.) apoptosis and cell cycle analysis (flow cytometry) in MDA-MB-468 and BT-20 cell lines. The optimal antiproliferative concentration of talazoparib and calcitriol in BT-20 was 91.6 and 10 µM, respectively, and in MDA-MB-468, it was 1 mM and 10 µM. Combined treatment significantly increased inhibition of cell migration in both cell lines. The combined treatment in BT-20 significantly increased late apoptosis (89.05 vs. control 0.63%) and S and G2/M populations (31.95 and 24.29% vs. control (18.62 and 12.09%)). Combined treatment in MDA-MB-468 significantly increased the S population (45.72%) and decreased G0/G1 (45.86%) vs. the control (26.79 and 59.78%, respectively). In MDA-MB-468, combined treatment significantly increased necrosis, early and late apoptosis (7.13, 33.53 and 47.1% vs. control (1.5, 3.1 and 2.83%, respectively)). Talazoparib and calcitriol combination significantly affected cell proliferation and migration, induction of apoptosis and necrosis in TNBC cell lines. This combination could be useful as a formulation to treat TNBC.


On leaving: Coloniality and physician migration in Puerto Rico.
Puerto Rico (PR) has a growing physician migration problem. As of 2009, the medical workforce was composed of 14,500 physicians and by 2020 the number had been reduced to 9,000. If this migration pattern continues, the Island will not be able to meet the recommended physicians per capita ratio proposed by the World Health Organization (WHO). Existing research has focused on the personal motivations for movement to, or permanence in, a particular setting, and social variables that encourage physicians to migrate (e.g., economic conditions). Few studies have addressed the role of coloniality in fostering physician migration. In this article we examine the role of coloniality and its impact on PR's physician migration problem. The data presented in this paper stem from an NIH-funded study (1R01MD014188) that aimed to document the factors associated with physician migration from PR to the US mainland and its impact on the Island's healthcare system. The research team used qualitative interviews, surveys, and ethnographic observations. This paper focuses on the data from the qualitative interviews with 26 physicians who had migrated to the USA and ethnographic observations, which were collected and analyzed between September 2020 and December 2022. The results evidence that participants understand physician migration as a consequence of three factors: 1) the historical and multidimensional deterioration of PR, 2) the idea that the current healthcare system is rigged by politicians and insurance companies, and 3) the specific challenges faced by physicians in training on the Island. We discuss the role of coloniality in fostering these factors and how it serves as the backdrop for the problem faced by the Island.


Ecophysiological adaptations to variable salinity environments in the crab Hemigrapsus crenulatus from the Southeastern Pacific coast: Sodium regulation, respiration and excretion.
The estuarine crab Hemigrapsus crenulatus is a key benthic species of estuarine and intertidal ecosystems of the South Pacific, habitats that experience wide fluctuations in salinity. The physiological strategies that allow this crab to thrive under variable salinities, and how they change during the benthic stages of their life cycle, were evaluated under laboratory conditions. Oxygen consumption, ammonia excretion and the regulatory capacity of Na+ through the normal range of environmental salinities (i.e. 5, 10, 15, 20, 25, 30) were evaluated in three size classes, ranging from juveniles to adults. In all sizes, the oxygen consumption, ammonia excretion and regulatory capacity of Na+ decreased as salinity increased, with the highest values at 5 and the lowest values at 30 salinity. Bigger crabs showed a higher capacity to regulate Na+, as well as higher respiration and excretion rates compared to smaller crabs, suggesting that they are better equipped to exploit areas of the estuary with low salinity. Regardless of its size, H. crenulatus is a strong hyper regulator in diluted media (i.e. 5-20) while a conformer at salinities higher than 20. The regulatory capacity of Na+ was positively related with oxygen consumption and ammonia excretion rates. These relationships between sodium regulation, respiration and excretion are interpreted as adaptive physiological mechanisms that allow H. crenulatus to maintain the osmotic and bioenergetic balance over a wide range of environmental salinities.


[No addiction in pain patients? Questions of a reading physician].
In some recent German textbooks of pain management there is maintained that psychological dependence to opioids in pain patients is not a real problem. The evidence for this statement is reviewed. Reading of the cited literature and additional MEDLINE search. Not a single convincing clinical study could be found in the references to proof the claimed absence of addiction in pain patients. The central topics of some papers cited had not been taken into account, other papers have been cited distorting their meaning or overemphasizing their true importance. Despite the assertion of the contrary, the maintenance of no addiction in pain patients, especially in noncancer pain, seems to be not yet proven.


GPR35 is a novel lysophosphatidic acid receptor.
GPR35 is a rhodopsin-like G protein-coupled receptor identified in 1998. It has been reported that kynurenic acid, a tryptophan metabolite, may act as an endogenous ligand for GPR35. However, the concentrations of kynurenic acid required to elicit the cellular responses are usually high, raising the possibility that another endogenous ligand may exist. In this study, we searched for another endogenous ligand for GPR35. Finally, we found that the magnitude of the Ca(2+) response induced by 2-acyl lysophosphatidic acid in the GPR35-expressing HEK293 cells was markedly greater than that in the vector-transfected control cells. Such a difference was not apparent in the case of 1-acyl lysophosphatidic acid. 2-Acyl lysophosphatidic acid also caused the sustained activation of RhoA and the phosphorylation of extracellular signal-regulated kinase, and triggered the internalization of the GPR35 molecule. These results strongly suggest that 2-acyl lysophosphatidic acid is an endogenous ligand for GPR35.


Antithrombin III (AT) and recombinant tissue plasminogen activator (R-TPA) used singly and in combination versus supportive care for treatment of endotoxin-induced disseminated intravascular coagulation (DIC) in the neonatal pig.
Disseminated intravascular coagulation (DIC) is a pathological disturbance of the complex balance between coagulation and anticoagulation that is precipitated by vascular injury, acidosis, endotoxin release and/or sepsis and characterized by severe bleeding and excessive clotting. The innately low levels of coagulation factors found in newborn infants place them at extremely high risk for DIC. Anecdotal reports suggest that either anticoagulant or fibrinolytic therapy may alleviate some of the manifestations of DIC. To test the hypothesis that replacement of both anticoagulants and fibrinolytics may improve survival and outcome better than either single agent or supportive care alone, we utilized a neonatal piglet model of endotoxin-induced DIC. DIC was induced in twenty-seven neonatal pigs (7 to 14 days of age) by intravenous administration of E. coli endotoxin (800 microg/kg over 30 min). The piglets were divided into 4 groups on the basis of treatment protocol [A: supportive care alone; B: Antithrombin III (AT, 50 microg/kg bolus, 25 microg/kg per hr continuous infusion) and supportive care; C: Recombinant Tissue Plasminogen Activator (R-TPA, 25 microg/kg per hr continuous infusion) and supportive care; D: AT, R-TPA and supportive care] and monitored for 3 primary outcome parameters (survival time, macroscopic and microscopic organ involvement) and 4 secondary outcome parameters (hematocrit; platelet count; fibrinogen level; and antithrombin III level). Compared with supportive care alone, combination therapy with AT and R-TPA resulted in a significant improvement of survival time, hematocrit, AT level, macroscopic and microscopic organ involvement, p < 0.05. Compared with supportive care alone, R-TPA alone significantly reduced macroscopic organ involvement and AT alone increased AT levels. The findings suggest that combining AT, R-TPA and supportive care may prove more advantageous in treating the clinical manifestations of DIC in this neonatal pig model than either single modality or supportive care alone.


Patterns of molecular variation. I. Interspecific comparisons of electromorphs in the Drosophila mulleri complex.
The average mobility of electromorphs at an enzyme locus in a single population was defined as the weighted average mobility of the electromorphs in that population, where the electromorph frequencies are the weights. A derivative distance measure was defined whose taxonomic utility was determined in the Drosophila mulleri species complex. Most of the variation in this metric was at the interspecific level, primarily among clusters of sibling species. The electromorphs of some loci were equally and regularly spaced, while those of other loci were less regular in their spacing. Overall, these minor perturbations from regular spacing did not noticeably detract from the taxonomic utility of average mobility, and cluster analysis yielded the same taxonomic relationships as more conventional nonmolecular treatments. On the other hand, electromorph spacing may be related to functional constraints on the enzyme molecules. Some possible implications of the results for the modes of selection during evolution of the different enzymes are discussed.


Comparative analysis of binding affinities to epidermal growth factor receptor of monoclonal antibodies nimotuzumab and cetuximab using different experimental animal models.
Although pharmaco/toxicological studies have always been conducted in pharmacologically relevant species in which the test material is pharmacologically active, the very specificity of many biopharmaceuticals could present challenges in the identification of a relevant species for pharmaco/toxicological studies. Alternative approaches may improve the predictive value of preclinical assessments of species-specific biopharmaceuticals. This could lead to improved decision-making, reduce the number of experimental animals by eliminating non-relevant studies, and decrease the time and cost involved in the drug development process. As an alternative to utilizing traditional animal models, this study investigated the activity of human EGF and the anti-EGF receptor monoclonal antibodies nimotuzumab and cetuximab using the placenta microsomal fraction of different experimental animals. Ligand-receptor binding curves were obtained from the different experimental animal models, and binding constants were calculated based on the Scatchard plots. The constants for human and monkey EGF receptor expressed on the placental extract showed a K(a)<10(-8)M, while rabbits, mice and rats showed a K(a)>10(-8)M. The K(a) values obtained from animal placentas show that Macaca fascicularis and Cercopitecus aethiops monkeys are relevant species for studying the pharmaco/toxicological properties of nimotuzumab and cetuximab.


Prostatic Artery Embolization Versus Transurethral Resection of the Prostate for Benign Prostatic Hyperplasia: A Cost-Effectiveness Analysis.
To compare the cost effectiveness of prostatic artery embolization (PAE) with that of transurethral resection of the prostate (TURP) for the treatment of medically refractory benign prostatic hyperplasia (BPH). A cost-effectiveness analysis with Markov modeling was performed, comparing the clinical course after PAE with that after TURP for 3 years. Probabilities were obtained from the available literature, and costs were based on Medicare reimbursements and published cost analyses. Outcomes were measured using quality-adjusted life-year (QALY). Statistical analyses included base case calculation, probabilistic sensitivity analysis, and deterministic sensitivity analysis to assess the robustness of the conclusion under different clinical scenarios. Base case calculation showed comparable outcomes (PAE, 2.845 QALY; TURP, 2.854 QALY), with a cost difference of $3,104 (PAE, $2,934; TURP, $6,038). The incremental cost-effectiveness ratio was $360,249/QALY. PAE was dominant in 23.2% and more cost effective in 48.4% of the probabilistic sensitivity analysis simulations. PAE was better if its recurrence risk was <20.4% per year and even when the TURP recurrence risk was assumed to be 0%. TURP would be more cost effective when its procedural cost was <$3,367 or the PAE procedural cost >$4,409. PAE remained cost effective when varying the risks and costs of the minor and major short-term or long-term adverse events of both procedures. TURP would be the better strategy if the utility of BPH recurrence was <0.85 QALY. PAE is a cost-effective strategy to treat medically refractory BPH, resulting in comparable health benefits at a lower cost than that of TURP even when accounting for extreme alterations in adverse events, costs, and recurrence rates.


Relation of single unit properties to the oculomotor function of the nucleus of the basal optic root (accessory optic system) in chickens.
Single unit recordings in the nucleus of the basal optic root (nBOR) of the accessory optic system in chickens suggest that it has a role in vertical stabilizing eye movements. Cells have unusually large receptive fields and never respond to small stationary stimuli. They respond best to large richly patterned stimuli moving slowly (2-4 degrees/s) in vertical directions. Cells responsive to upward movement tend to be located in the dorsal portion of nBOR, which projects to motor areas producing upward eye movement, whereas cells responsive to downward movement tend to be located in the ventral portion of nBOR, which projects to motor areas producing downward eye movement; this suggests that these synapses onto oculomotor neurons are excitatory. In many nBOR units, the preferred and null directions are not opposite to each other. These directional asymmetries seem to be correlated with other properties of the units in a manner that supports the idea that the accessory optic system is arranged according to a vestibular coordinate system. This finding complements the abundant anatomical and physiological evidence linking the accessory optic system to the vestibular system.


Identification of lymphocytes in percutaneous liver biopsy cores. Different T:B cell ratio in HB sAg-positive and -negative hepatitis.
The lymphocytes infiltrating the liver were isolated and characterized as T or B cells in three groups of patients: 20 patients with hepatitis B surface antigen (HB sAg)-positive acute and chronic hepatitis, 8 patients with HBsAg-negative chronic hepatitis with prior evidence for hepatitis B virus (HBV) infection, and 5 patients with HBsAg-negative chronic hepatitis without prior evidence for HBV infection. The predominant cell infiltrating the liver was shown to be a T cell in all categories; however, the ratio of T:B cells was significantly lower (1.96) in the patients without evidence for HBV infection than in the patients who were HBsAg-positive at (7.86), or before (8.85) the time of study. The significantly (P less than 0.001) higher number of B cells in the patients with chronic hepatitis of unknown etiology suggests that a different immunopathogenetic mechanism is operative in this group. A peripheral T lymphocytopenia was observed in patients with both antecedent and existent HBs-antigenemia, but not in the patients without evidence for HBV infection.


An improved method for the concentration of cerebrospinal fluid cells by suction tip and sedimentation chamber.
To concentrate cells from cerebrospinal fluid, a new method was developed. The technique is a modification of the cell sedimentation method of Sayk. The fluid is absorbed after passing through a Nuclepore filter, pore size 0.4 micron. With the aid of simple utensils it is possible to obtain a cell preparation of high quality using only 0.3 ml of cerebrospinal fluid. The cytomorphology is as good as that achieved by Sayk's sedimentation technique. The cell yield was determined using an accurate and precise method to estimate the number of cells present in the cerebrospinal fluid. The average cell yield is 90%, and all staining methods are applicable. The system is easily modified, and can be suited to many other fluids and purposes.


Cell-free protein synthesis of perdeuterated proteins for NMR studies.
Cell-free protein synthesis protocols for uniformly deuterated proteins typically yield low, non-uniform deuteration levels. This paper introduces an E. coli cell-extract, D-S30, which enables efficient production of proteins with high deuteration levels for all non-labile hydrogen atom positions. Potential applications of the new protocol may include production of proteins with selective isotope-labeling of selected amino acid residues on a perdeuterated background for studies of enzyme active sites or for ligand screening in drug discovery projects, as well as the synthesis of perdeuterated polypeptides for NMR spectroscopy with large supra-molecular structures. As an illustration, it is demonstrated that the 800-kDa chaperonine GroEL synthesized with the D-S30 cell-free system had a uniform deuteration level of about 95% and assembled into its biologically active oligomeric form.


Dynamics of Na,K-ATPase sites in lateral cochlear wall tissues of the rat.
The objective of this study was to determine whether varying levels of either the synthetic glucocorticoid, dexamethasone, or the mineralocorticoid, aldosterone, modulate the quantity of Na,K-ATPase sites in stria vascularis and spiral ligament tissues. Surgically adrenalectomized male rats were administered different dosages of dexamethasone or aldosterone for 7 days and subsequently were sacrificed. Na,K-ATPase alpha-subunits of stria vascularis and spiral ligament homogenates were determined quantitatively by enzyme-linked immunosorbent assay, using a monoclonal antibody shown to react with lateral wall Na,K-ATPase alpha-subunit. Elevated serum levels of dexamethasone were correlated with significantly increased Na,K-ATPase alpha-subunit levels in both the stria vascularis and spiral ligament (P < 0.05). Elevated serum levels of aldosterone were correlated with increased Na,K-ATPase alpha-subunits in the stria vascularis, but not in the spiral ligament (P < 0.1). These data further indicate a positive correlation between increased serum levels of adrenal steroids and induction of Na,K-ATPase synthesis by such steroids, particularly by dexamethasone.


Network percolation provides early warnings of abrupt changes in coupled oscillatory systems: An explanatory analysis.
Functional networks are powerful tools to study statistical interdependency structures in spatially extended or multivariable systems. They have been used to get insights into the dynamics of complex systems in various areas of science. In particular, percolation properties of correlation networks have been employed to identify early warning signals of critical transitions. In this work, we further investigate the corresponding potential of percolation measures for the anticipation of different types of sudden shifts in the state of coupled irregularly oscillating systems. As a paradigmatic model system, we study the dynamics of a ring of diffusively coupled noisy FitzHugh-Nagumo oscillators and show that, when the oscillators are nearly completely synchronized, the percolation-based precursors successfully provide very early warnings of the rapid switches between the two states of the system. We clarify the mechanisms behind the percolation transition by separating global trends given by the mean-field behavior from the synchronization of individual stochastic fluctuations. We then apply the same methodology to real-world data of sea surface temperature anomalies during different phases of the El Niño-Southern Oscillation. This leads to a better understanding of the factors that make percolation precursors effective as early warning indicators of incipient El Niño and La Niña events.


Is vaccination against IgE possible?
A substantial reduction in the levels of both total and antigen specific IgE will most likely result in improved symptom scores in atopic individuals. Based on this assumption we initiated a project to study the possibility of reducing levels of circulating and mast cell bound IgE, by inducing a strong autoimmune antibody response against IgE in the host. Bacterially produced fusion proteins containing constant domains two (CH2) and three (CH3) of rat IgE directly linked to the glutathione-S-transferase (GST) protein from Schistosoma japonicum or to the maltose binding protein of Esherichia coli were used as the active components of the allergy vaccine. Injection of either of these fusion proteins together with adjuvant led to the induction of a strong autoimmune anti-IgE response in several IgE low or medium responder strains of rats. Vaccination of ovalbumin sensitised Wistar rats with the GST-C2C3 fusion protein resulted in a profound decrease in serum IgE levels and later in a nearly complete block in histamine release from mast cells and basophils upon challenge with either a cross-linking polyclonal anti-IgE antiserum or a specific allergen. This shows that it is possible to reduce IgE levels in an animal to such an extent that it gives a clear clinical effect. Recent studies with an extended panel of rat strains including four IgE high responder strains, indicate that induction of the autoimmune response is dependent on the plasma concentration of IgE before vaccination. A high concentration of IgE has a negative effect on the induction of autoimmunity, most likely by inducing a B-cell tolerance in the host. Vaccinated subjects with very high IgE concentrations thereby responds poorly to the vaccine. Current studies are aimed at overcoming this potential limitation of the vaccination procedure.


Relationships between the nuclear membrane, nuclear pore complexes, and organelles in the type II pneumocyte.
Functional relationships among organelles of the type II cell are suggested based upon the proximity of organelles to specialized areas of the plasma- and nuclear membranes. In a two-dimensional morphometric analysis of the profiles of organelles in type II cells of the ferret and rat (and beagle dog), lamellar bodies were more likely to be located near the nuclear membrane than at the alveolar space (where exocytosis occurs). The size of lamellar body profiles was not correlated with distance from the nuclear membrane; however, large profiles were nearer the apical membrane, and smaller ones nearer the basement membrane. Profiles of highly branched mitochondria were 10 times more frequently associated with nuclear pore complexes than with the inter-pore nuclear membrane. Forty percent of all mitochondrial profiles were within 0.25 microns of the nucleus, 5% were within 0.02 microns and half of these appeared to touch the filaments of the nuclear pore complexes. The size of mitochondrial profiles was not correlated with distribution. In the ferret and rat, 8.6% and 2.5% respectively, of the nuclear pore complexes were associated with mitochondria. Sebaceous cells, from control mice, demonstrated a spatial distribution of granules which was size dependent but unrelated to the nuclear membrane.


Group A rotavirus prevalence and genotypes among adult outpatients with diarrhea in Beijing, China, 2011-2018.
Group A rotavirus (RVA) is one of the most common causes of severe diarrhea in children worldwide. However, RVA is also an important pathogen causing adult diarrhea, with higher infection rates in older patients. To provide evidence for rotavirus epidemic control and to inform vaccine development, we analyzed the molecular epidemiology of RVA among adult outpatients with diarrhea in Beijing from 2011 to 2018. Stool specimens were collected monthly from 14 districts. RVA was detected using enzyme-linked immunosorbent assay and real-time reverse-transcription polymerase chain reaction (RT-PCR). Genotyping of rotavirus was performed using multiplex semi-nested RT-PCR. Phylogenetic analysis was performed using maximum likelihood methods implemented in MEGA software (version 6.06). Logistic regression and chi-square tests were used to assess differences among age groups, districts, years, and genotype distributions. The prevalence of rotavirus was 10.16% (1310 of 12,893) among adult outpatients with diarrhea from 2011 to 2018 in Beijing. The highest prevalence (13.74%, 600 of 4367) was observed among those aged 41 to 65 years. November, December, and January had the highest positive detection rates. In 2011, G3P[8] and G9P[8] were the dominant genotypes. Starting from 2012, G9P[8] became the dominant genotype. Most G9 strains belonged to the G9-VI clade. Most P[8] strains belonged to the P[8]-III clade. RVA is a major cause of adult diarrhea in Beijing. Continuous molecular surveillance is needed, and transmission of rotavirus between children and adults should be investigated further.


Assessment of indoor allergen exposure.
Of the four modes of treating human allergic disease, avoidance or separation of the allergic patient from the allergen source is most effective and least expensive. The clinical immunology laboratory has established efficient and inexpensive "reservoir" dust sampling and processing procedures to obtain a surface dust specimen that reflects the allergen burden of the environment. Following extraction, allergens are quantified by reproducible, validated immunoenzymetric assays for the quantification of "indicator" aeroallergen levels in home, school, and work environments. In this paper, the strategies and methods for collecting and processing dust samples are discussed, and assays are reviewed for quantifying indoor aeroallergen exposure from dust mites (Der p 1 and 2, Der f 1 and 2), animals (cat: Fel d 1; dog: Can f 1; mouse: Mus m 1; rat: Rat n 1), and insects (cockroach: Bla g 1 and 2). Accurate quantification of the levels of allergen in indoor environments facilitates avoidance therapy by identifying environmental risk factors for asthma and allergy exacerbation and allowing the allergic patient to monitor the effectiveness of environmental remediation actions.


Intrauterine tranfusion: 101 consecutive cases treated at Queen Charlotte's Maternity Hospital.
Intrauterine transfusion was performed or attempted in 101 cases during a period of four and a half years. After exclusion of six cases, for reasons detailed in the text, 95 cases are considered in detail. Intrauterine transfusion was successfully performed in 93 of these, with a mean of 2.8 transfusions each. Forty-four babies (46.3%) survived and all but one are so far developing normally.