Effect of composition and different fractions of hay dust suspension on inflammation in lungs of heaves-affected horses: MMP-9 and MMP-2 as indicators of tissue destruction.
Airway matrix metalloproteinases (MMPs) increase following inhalation of organic dust. The relative contribution of dust components to this elevation is unknown. To identify components of organic dust responsible for elevated MMP levels in equine airways. Bronchoalveolar lavage (BALF) from 7 heaves-susceptible horses, collected 6 h following inhalation challenges with saline, 2 different hay dust suspensions (HDS-1 and -2) and soluble and particulate fractions of HDS-1, were analysed for MMP-2 and -9 using SDS-page gelatin zymography. HDS-1 challenge increased BALF proMMP-9 and total MMP-9. HDS-1 fractions, or the particulate fraction with added lipopolysaccharide, increased BALF proMMP-9 and total MMP-9 in combination, but not when inhaled separately. HDS-2 inhalation elevated BALF complex forms, proMMP-9, active MMP-9, total MMP-9 and total MMP-2. The results suggest synergistic action of soluble and particulate organic dust components. The fact that HDS-1 and HDS-2 had different glucan concentrations supports a role for moulds in the activation of MMP-9. Activation and release of MMPs in response to inhaled moulds are involved in the aetiopathogenesis of heaves. Endotoxin contributes to the synergistic action of the dust components, but the overall MMP response to organic dust inhalation in heaves-susceptible horses largely reflects the mould content of the dust. In the future, inhibition of MMP production and release may offer therapeutic means for treatment and prevention of heaves and recommendations for acceptable dust levels can be given.


Development of preservative-free nanoparticles-based emulsions: Effects of NP surface properties and sterilization process.
Model emulsions were developed with or without commercial titanium dioxide nanoparticles (NP) carrying various surface treatments in order to get close physicochemical properties whatever the NP surface polarity (hydrophilic and hydrophobic). Rheology and texturometry highlighted that the macroscopic properties of the three formulated emulsions were similar. However, characterizations by optical microscopy, static light scattering and zetametry showed that their microstructures reflected the diversity of the incorporated NP surface properties. In order to use these model emulsions as tools for biological evaluations of the NP in use, they had to show the lowest initial microbiological charge and, specifically for the NP-free emulsion, the lowest bactericidal effect. Hence, formulae were developed preservative-free and a thermal sterilization step was conducted. Efficiency of the sterilization and its impact on the emulsion integrity were monitored. Results highlighted the effect of the NP surface properties: only the control emulsion and the emulsion containing hydrophilic NP fulfilled both requirements. To ensure the usability of these model emulsions as tools to evaluate the 'NP effect' on representative bacteria of the skin microflora (S. aureus and P. fluorescens), impact on the bacterial growth was measured on voluntary inoculated formulae.


Rate of tryptophan hydroxylation in vivo in brain nuclei of genetically hypertensive rats of the Lyon strain.
The rate of tryptophan hydroxylation in vivo is unaltered in brain areas of 5, 9 and 21 week-old Lyon genetically Hypertensive (LH) rats as compared to both Lyon Normotensive (LN) and Low Blood Pressure (LL) rats, except for a decrease in the C1 area of the medulla oblongata in 9 week-old animals.


Model-based mid-infrared spectroscopy for on-line monitoring of volatile fatty acids in the anaerobic digester.
The performance of anaerobic digestion is significantly governed by the concentration of volatile fatty acids (VFAs). Though the titration and near-infrared spectroscopy have been used to measure the VFAs in the digester, there is still lack of the establishment of on-line monitoring of VFAs in practical application. An effective quantification method based on mid-infrared (MIR) spectroscopy was developed, and used to measure the concentrations of VFAs in the anaerobic bioreactor nondestructively in parallel. The wavelet denoising (WD) spectra were used as the spectral preprocessing option. Compared with other pretreatment methods, the established calibration model built by WD spectra showed satisfactory results. Further, the model was verified using high performance liquid chromatography (HPLC), and predictions were made using real reactor effluent samples. Based on this theoretical work, a set of equipment for the in-situ online monitoring of VFAs was designed, which has high feasibility and effectively solves the problems with the current VFAs online monitoring process. These results provide a new solution for on-line monitoring of the anaerobic digestion, and have great potential for practical application.


Methods of outbreak investigation in the "era of bacteriology" 1880-1920.
The advent of bacteriological methods in the later 19th century has been seen, on the examples of America and Germany, to have been followed by a new laboratory-based, contact-tracing method of investigating outbreaks of epidemic disease. In Britain, however, this new approach never took firm root, and practising epidemiologists continued to follow an observational and deductive tradition in field investigations, rejecting any primary dependence on bacteriological methods. Alongside this persistent observational practice, there emerged a new statistical approach, based in Pearsonian biometrics, which allied itself with experimental laboratory techniques to develop a more systematic, theoretical trajectory for explaining disease outbreaks in the years after World War I.


Characterization of a prophenoloxidase from hemocytes of the shrimp Litopenaeus vannamei that is down-regulated by white spot syndrome virus.
Previously, a prophenoloxidase (proPO) gene (named proPO-a here) from hemocytes of Litopenaeus vannamei was isolated. Here, a proPO-b gene was also identified and characterized from hemocytes of L. vannamei. The cDNA sequences of proPO-a and proPO-b were compared, and it was found that both proPOs had a microsatellite DNA site near the 3' end of the open reading frame (ORF). However, the microsatellite DNA of proPO-b contained a compound imperfect simple sequence repeats (SSR) ((CT)(38)(CA)(8)(AA)(CA)(3)(TA)(CA)(14)), which was different from the perfect one ((CT)(20)) of proPO-a, and the cDNA sequences of proPO-a and proPO-b prior to the microsatellite DNA were almost identical, but differed after the microsatellite DNA. ProPO-b (3232 bp) was longer than proPO-a (2471 bp). The 3' UTR sequence after SSR of proPO-a was not detected in shrimp randomly collected from five different geographically separate populations by reverse-transcription polymerase chain reaction (RT-PCR). On the contrary, the 3' UTR sequence of proPO-b was detected in all five groups of shrimps. Northern blot analysis showed that a transcript at approximately 3.2kb, but not 2.5kb, was detected mainly in hemocytes, and also present in midgut, gill, heart, stomach, posterior midgut cecum, and cuticular epidermis, but no signal was detected in hepatopancreas and musculature. RT-PCR and quantitative real-time RT-PCR analysis showed similar results of the proPO-b expression profile in these shrimp tissues. We also observed that proPO-b expression was down-regulated in shrimp challenged with white spot syndrome virus (WSSV). Our results suggest that proPO-b is a main transcript form of proPO gene in L. vannamei, and it may play a role in defence against WSSV virus.


Pulmonary arterial hypertension: use of delayed contrast-enhanced cardiovascular magnetic resonance in risk assessment.
Pulmonary arterial hypertension is a severe and progressive disease. Its early diagnosis is the greatest clinical challenge. To evaluate the presence and extension of the delayed myocardial contrast-enhanced cardiovascular magnetic resonance, as well as to verify if the percentage of the myocardial fibrosis mass is a severity predictor. Cross-sectional study with 30 patients with pulmonary arterial hypertension of groups I and IV, subjected to clinical, functional and hemodynamic evaluation, and to cardiac magnetic resonance. The mean age of patients was 52 years old, with female predominance (77%). Among the patients, 53% had right ventricular failure at diagnosis, and 90% were in functional class II/III. The mean of the 6-minute walk test was 395 m. In hemodynamic study with right catheterism, the mean average pulmonary arterial pressure was 53.3 mmHg, of the cardiac index of 2.1 L/min.m(2), and median right atrial pressure was 13.5 mmHg. Delayed myocardial contrast-enhanced cardiovascular magnetic resonance was found in 28 patients. The mean fibrosis mass was 9.9 g and the median percentage of fibrosis mass was 6.17%. The presence of functional class IV, right ventricular failure at diagnosis, 6-minute walk test < 300 meters and right atrial pressure > 15 mmHg, with cardiac index < 2.0 L/min.m(2), there was a relevant association with the increased percentage of myocardial fibrosis. The percentage of the myocardial fibrosis mass indicates a non-invasive marker with promising perspectives in identifying patients with high risk factors for pulmonary hypertension.


Delayed greening, leaf expansion, and damage to sympatric Shorea species in a lowland rain forest.
The function of delayed greening in the seedlings of canopy tree species in a lowland tropical rain forest was examined in terms of its potential defensive value against herbivory. To explore the ecological and evolutionary backgrounds for delayed greening, we chose eight sympatric congeneric (Shorea) dipterocarp species that were either normal-greening or delayed-greening species. Expansion and toughening of leaves took approximately 30 days for all species, and did not differ between the normal- and delayed-greening species. The main factors that affected leaf damage during expansion were insect herbivory and fungal infection. Levels of leaf damage were significantly lower for delayed-greening species than for normal-greening species, but proportions of heavily damaged leaves and leaf abscission during expansion did not differ. In addition, no significant difference was found in damage levels on leaves (aged 1-2 months) of naturally occurring seedlings between normal- and delayed-greening species. Therefore, delayed greening may effectively reduce the level of leaf damage in young expanding leaves, but may not necessarily reduce leaf abscission and damage to mature leaves. The existence of delayed greening could not be simply explained by the phylogenetic and ecological backgrounds of the trees. Consequently, delayed greening may have a function in reducing damage during expansion, but more information (such as knowledge of the secondary metabolites involved in this phenomenon) is needed to explain fully why these species exhibit delayed greening.


Association between per- and polyfluoroalkyl substances and semen quality.
Some studies have suggested that perfluoroalkyl and polyfluoroalkyl substance (PFAS) exposure may be associated with semen quality in the general population, but with inconsistent results. To identify a more precise relationship between them, a meta-analysis was performed. We searched Embase, the PubMed, The Cochrane Library, Ovid databases, and Web of Science databases (before March 2022) for appropriate studies on the correlations of PFAS exposure with semen parameters. We extracted β value and 95% confidence intervals (CIs) to conduct meta-analysis. Subgroup analyses was performed by sample size, geographic location, and sample type. A total of seven articles involving 2190 participants were included in this study. The concentrations of perfluorooctanoic acid (PFOA) (β value = - 1.38; 95% CI: - 2.44, - 0.32) and perfluorononanoic acid (PFNA) (β value = - 1.31, 95% CI: - 2.35, - 0.26) were negatively associated with sperm progressive motility. Subgroup analysis revealed that PFNA exposure was related to sperm morphology in studies with the sample size exceeding 200 people (β value = - 0.14; 95% CI: - 0.26, - 0.01). Our study supports that exposure to some PFASs (e.g., PFNA, PFOA) may be associated with semen quality, such as lower sperm progressive motility. Therefore, it is of great significance for the prevention of male infertility by control the use of PFASs.


[Retroperitoneal tumors in children: clinical diagnostic problems (author's transl)].
Starting from two recent observations of retroperitoneal tumors in the pediatric age group (a nephroblastoma and a ganglioneuroma), the authors illustrate the main characteristics of these malignancies and emphasize the difficulties often encountered in differential diagnosis.


Morphometry of the pyramidal tract in the spinal cord of normal rats.
A morphometric study of the pyramidal tract at the spinal cord level in the normal albino rat is reported. The area, perimeter, axonal and glial cell count, and axonal concentration were determined at the seventh cervical, second thoracic, and third lumbar spinal cord levels. The results show consistent diminishing values of the area, axonal and glial cell count, and axonal concentration with the tract descent in the spinal cord.


Dual-jet electrospun PDLGA/PCU nonwovens and their mechanical and hydrolytic degradation properties.
A dual-jet electrospinning was used to mix a different hydrophilicity poly(carbonate urethanes) (PCUs) nanofibers with a biodegradable poly(D,L-lactide-co-glycolide) (PDLGA) copolyester microfibers. As a result, PDLGA/PCU partially degradable nonwovens consisting of an interlaced of both components fibers were obtained. In order to examine the hydrolytic degradation process of polyester fraction, as well as changes that occurred in the mechanical properties of the whole nonwovens, gel permeation chromatography, proton nuclear magnetic resonance spectroscopy, differential scanning calorimetry and scanning electron microscopy as well as static tensile test were performed. Obtained results showed that for the introduction of more hydrophobic PCU nanofibers (ChronoSil), the process of copolyester chain scission slowed down and the erosion mechanism proceeded in bulk. Unexpectedly, even greater deceleration of PDLGA fibers degradation was observed in case of more hydrophilic PCU (HydroThane), and erosion mechanism changed to surface. Enhancement the affinity of the whole nonwoven to the water, manifested by strong water uptake, facilitated the diffusion processes of both: water and acid degradation by-products, which limited autocatalysis reactions of the hydrolysis of ester bonds. On the other hand, strength tests showed the synergy in the mechanical characteristics of both components. Presented method allows influencing the mechanism and rate of polyester degradation without changing its chemical composition and physical properties, affecting only the physical interactions between the nonwoven and the degradation environment, and thus, on diffusion processes. Obtained partially degradable materials possessed also time prolonged functional properties, compared to the copolyester-only nonwoven itself, thus could be considered as promising for biomedical applications e.g. in drug release systems, implants or surgical meshes for supporting soft tissues.


Gene cloning, expression and functional characterization of a proliferation-inducing ligand (APRIL) from hedgehog (Erinaceus europaeus).
Here we describe the identification of the hedgehog Erinaceus europaeus homologue of a proliferation-inducing ligand (APRIL) of the TNF family (designated heAPRIL). Hedgehog APRIL contains two cysteine residues (Cys(196) and Cys(211)), a furin protease cleavage site and a conserved putative N-glycosylation site (Asn(124)). Real-time quantitative PCR (qPCR) analysis revealed that heAPRIL could be detected in various tissues. MTT assays and flow cytometric analysis revealed that Nus-hesAPRIL and hesAPRIL could promote the survival/proliferation of splenic B cells. Laser scanning confocal microscopy analysis showed GFP-hesAPRIL could successfully bind to the APRIL receptors of lymphocytes.


[Adhesion of glass ionomer to composite resins. Several practical data].
The traction resistance of the bond between vitreous ionomer and composite after acid treatment of the former is analysed using various types of composite.


Influence of extreme hypercapnia on respiratory motor nerve activity in cats.
Sedative drugs have been found to depress the respiratory activity of upper airway muscles more than that of the diaphragm. To determine whether CO2 at narcotic levels has a similar action, we recorded phrenic and hypoglossal nerve activities in decerebrate, vagotomized, paralyzed cats. T5 or T6 external intercostal nerve activity was also recorded in some animals. End-tidal CO2 concentration was raised progressively to over 30% or until depression of nerve activity was apparent. Respiratory frequency was reduced by severe hypercapnia in most cats. Hypoglossal nerve activity was consistently decreased more than that of the phrenic nerve. In most cases intercostal nerve activity was also more susceptible than phrenic nerve activity to hypercapnic depression. The results indicate that CO2 at narcotic levels interferes both with the central pattern generator for breathing movements and with the expression of the pattern in specific motor nerves.


Proteoglycan isolated from Phellinus linteus activates murine B lymphocytes via protein kinase C and protein tyrosine kinase.
Medicinal mushrooms are increasingly used to treat a wide variety of disease processes. Aqueous extract from the fruiting body or mycelia of Phellinus linteus has been reported to produce antitumor and immunomodulatory activities in vivo and in vitro. However, the therapeutic mechanism has not been known. In the present study, we investigated whether proteoglycan (PL) isolated from P. linteus has an effect on the immunomodulatory activities of the murine splenic lymphocytes (MSLs). Treatment with PL caused a four-fold augmentation in [3H]thymidine incorporation compared to untreated control group in MSLs. Flow cytometric analysis indicated that the affected cell population was mainly CD19(+) cells, but not CD3(+) cells. These data suggested that the main target of PL was the B cells, but not T cells. PL also enhanced the expression of co-stimulatory molecules, CD80 and CD86, in murine B cells in a time-dependent manner. Accordingly, we investigated if intracellular [Ca(2+)] and reactive oxygen intermediates (ROI) were the principal downstream components that contributed to PL-induced activation, with respect to both increases of proliferation and induction of co-stimulatory molecules. However, PL has no influence on the [Ca(2+)] concentration and the ROI formation in murine B cells, whereas the genistein, protein tyrosine kinase (PTK) inhibitor or staurosporine, protein kinase C (PKC), blocked the proliferation and the induction of co-stimulatory molecules, CD80 and CD86, in B cells stimulated with PL. Taken together, these data suggest that PL is a biological response modifier that stimulates proliferation and expression of co-stimulatory molecules in B cells, probably by regulating PTK and PKC signaling pathways.


Serine proteases identified from a Costelytra zealandica (White) (Coleoptera: Scarabaeidae) midgut EST library and their expression through insect development.
Costelytra zealandica larvae are pests of New Zealand pastures causing damage by feeding on the roots of grasses and clovers. The major larval protein digestive enzymes are serine proteases (SPs), which are targets for disruption in pest control. An expressed sequence tag (EST) library from healthy, third instar larval midgut tissue was constructed and analysed to determine the composition and regulation of proteases in the C. zealandica larval midgut. Gene mining identified three trypsin-like and 11 chymotrypsin-like SPs spread among four major subgroups. Representative SPs were examined by quantitative PCR and enzyme activity assayed across developmental stages. The serine protease genes examined were expressed throughout feeding stages and downregulated in nonfeeding stages. The study will improve targeting of protease inhibitors and bacterial disruptors of SP synthesis.


Effect of the GnRH stimulation test on Canine Prostatic-Specific Esterase (CPSE).
The gonadotropin-releasing hormone (GnRH) stimulation test is used to investigate testicular production of testosterone (T) when performing a breeding soundness examination. In male dogs with fertility problems, the prostate should also be investigated as prostatic conditions may frequently lower semen quality. Serum concentrations of canine prostatic-specific esterase (CPSE) increase in dogs with benign prostatic hyperplasia (BPH). When performing a breeding soundness examination in a male dog, GnRH administration is frequently done at the beginning of the process and then both T and CPSE are assayed on the same serum sample collected 1 h following the GnRH injection. The aim of this study was to assess whether or not the administration of GnRH may alter CPSE concentrations in dogs with a healthy prostate. Twenty-eight client-owned intact adult male dogs were included in the study. Following a 7-day sexual rest all male dogs underwent a clinical examination and an ultrasonographic examination of the prostatic gland. Prostatic size and parenchyma of every tested dog were evaluated by ultrasonography to assess prostatic conditions. Two different GnRH stimulation protocols were used, A = gonadorelin 50μg/dog SC (n = 15) and B = buserelin 0.12 μg/kg IV (n = 13). T and CPSE concentrations were measured before and 1 h after GnRH administration by a laser-induced fluorescence analysis. Buserelin and gonadorelin were equally effective in causing a significant increase in serum T concentrations in the post GnRH sample. When considering the 28 dogs together, CPSE concentrations did not change following the stimulation test with either GnRH compound; however, in 4/28 cases, the post GnRH value was markedly increased to values compatible with a diagnosis of BPH. There was no difference in the action of buserelin or gonadorelin in causing an increase in serum T concentrations. CPSE secretion was increased in approximately 15% of dogs treated with either buserelin or gonadorelin. Therefore, whenever performing diagnostic testing in intact male dogs, CPSE should not be assayed on a post-GnRH serum sample.


Evidence that human autoimmune thrombocytopenia mediated by both immunoglobulin isotypes IgM and IgG is an independent disease entity.
Autoimmune thrombocytopenic purpura (AITP) is a bleeding disorder caused by clonally restricted self-reactive antibodies with specificity for platelet glycoproteins. Anti-platelet autoantibodies in AITP mainly belong to the IgG class. The occurrence of anti-platelet autoantibodies of the IgM isotype has been reported, and AITP is partially mediated by antibodies of both isotypes, IgM and IgG. Using a technique of quantitative immunoblotting of immunoglobulins on self-tissues, followed by multiparametric statistical analysis of the data, we here demonstrate that patients with IgM- and IgG-mediated AITP are readily discriminated from patients with IgM-mediated AITP as well as from patients with IgG-mediated AITP at the basis of self-reactive antibody repertoires of isotypes IgM and IgG toward non-platelet antigens of human origin. Our data suggest that, in view of the important physiological functions of self-reactive antibody repertoires, human AITP mediated by both immunoglobulin isotypes IgG and IgM may be an independent disease entity. The role of autoantibody isotype for the pathophysiology of AITP might currently be underestimated, and diagnostic and therapeutic procedures in AITP might profit from considering autoantibody isotype more carefully.


Memory for reward location is enhanced even though acetylcholine efflux within the amygdala is impaired in rats with damage to the diencephalon produced by thiamine deficiency.
A rodent model of diencephalic amnesia produced by thiamine deficiency (pyrithiamine-induced thiamine deficiency [PTD]) was implemented to assess both changes in behavior and acetylcholine (ACh) efflux in the amygdala across four training sessions of a delayed alternation task. Two versions of the delayed alternation task were used. In one version, when a correct alternation was made a unique reward was paired with each spatial location ([left arm-chocolate milk] or [right arm-rat chow]). This paradigm is called the differential outcomes procedure (DOP). In the second version of the task, correct delayed alternation resulted in the same rewards but randomized across location (Nondifferential Outcomes Procedure [NOP]). The PTD rats were impaired on the first session of delayed alternation testing. However, both control and PTD rats using the DOP performed significantly better on delayed alternation than rats trained with the NOP.This effect was driven primarily by the PTD rats in the DOP condition outperforming all other groups on sessions 2-4. Although ACh efflux in the amygdala increased during delayed alternation testing in all groups, the NOP-trained rats had a greater rise in training-related ACh release in the post-training period. This suggests that increased amygdalar cholinergic activation is more critical for processing spatial information than episodic reward information. These data correspond with the idea that cholinergic activation of the amygdala promotes processing in other neural systems.


Incidence and consequence of close margins in patients with ductal carcinoma-in situ treated with mastectomy: is further therapy warranted?
The impact of close margins in patients with ductal carcinoma-in situ (DCIS) treated with mastectomy is unclear; however, this finding may lead to a recommendation for postmastectomy radiotherapy (PMRT). We sought to determine the incidence and consequences of close margins in patients with DCIS treated with mastectomy. The records of 810 patients with DCIS treated with mastectomy from 1996 through 2009 were reviewed. Clinical and pathologic factors were analyzed with respect to final margin status. Median follow-up was 6.3 years. Overall, 94 patients (11.7 %) had close margins (positive, n = 5; negative but ≤1 mm, n = 54; 1.1-2.9 mm, n = 35). Independent risk factors for close margins included multicentricity, pathologic lesion size ≥1.5 cm, and necrosis, but not age, use of skin-sparing mastectomy, or immediate reconstruction (p > 0.05). Seven patients received PMRT, and none had a locoregional recurrence (LRR). Among the remaining 803 patients, the 10-year LRR rate was 1 % (5.0 % for margins ≤1 mm, 3.6 % for margins 1.1-2.9 mm, and 0.7 % for margins ≥3 mm [p < 0.001]). The 10-year rate of contralateral breast cancer was 6.4 %. On multivariate analysis, close margins was the only independent predictor of LRR (p = 0.005). Close margins occur in a minority of patients undergoing mastectomy for DCIS and is the only independent risk factor for LRR. As the LRR rate in patients with close margins is low and less than the rate of contralateral breast cancer, PMRT is not warranted except for patients with multiple close/positive margins that cannot be surgically excised.


Generalized versus performance-focused social phobia: patterns of comorbidity among youth.
The goal of this study was to examine patterns of comorbidity associated with generalized and performance-focused social phobia (SOP) among a community-based sample of youth while also examining whether gender and age influenced the associations found. Participants from the Methods for the Epidemiology of Child and Adolescent Mental Disorders (MECA) Study, aged 9-17 (N=1295), were used. Diagnoses were based on structured clinical interviews with youth and their caregivers. Results indicated that both subtypes of SOP were associated with a range of other anxiety disorders. Contrary to the expectations, performance-focused SOP was somewhat more strongly associated with depressive disorders than was generalized SOP; this difference was driven primarily by higher rates of dysthymia (DYS) among those with performance-focused SOP than generalized SOP. Although attention-deficit hyperactivity disorder (ADHD) was associated with generalized SOP, more severe externalizing behavior (oppositional defiant disorder (ODD) and conduct disorder (CD)) was only associated with performance-focused SOP. Limited gender and age differences in these relationships were found.


Studies on the interactions of AFPs and bZIP transcription factor ABI5.
AFP1 interacts with ABI5 and negatively regulates the abscisic acid (ABA) signaling by accelerating ABI5's degradation during the seed germination phase in Arabidopsis, but the underlying mechanism remains unclear. Moreover, the molecular basis of the interaction between AFP1 homologs and ABI5 has yet to be elucidated. In this study, the patterns of their interactions with ABI5 were investigated in detail. We found that AFP2/3/4 can bind two regions of ABI5, one is ABI51aa to 135aa and another is ABI5202aa to 213aa. However, AFP1 only interacts with the second region of ABI5, i.e. ABI5202aa to 213aa. Prior research has shown that ABI51aa to 135aa is related to the transcriptional activity of ABI5. Thus, our results suggest that AFPs may also modulate ABI5, by directly binding to its transcriptional activation domain, thereby influencing its transcriptional activity. Further, interactions between AFPs and ABI5 were not affected if the Ser42th in the ABI5-SnRK2 motif were mutated respectively to Glu or Ala. Nevertheless, interactions between AFPs and ABI5 were eliminated if the Thr47th and Thr206th of ABI5 were mutated respectively to Glu or Ala. Since the two residues of Thr47th and Thr206th were located in the phosphorylation motifs of CKII, AFPs might regulate the activities of ABI5 transcription factor through a CKII-dependent pathway.


Epigenetics: a promising paradigm for better understanding and managing pain.
Epigenetic regulation of gene expression is a rapidly growing area of research. Considering the longevity and plasticity of neurons, the studies on epigenetic pathways in the nervous system should be of special interest for both epigeneticists and neuroscientists. Activation or inactivation of different epigenetic pathways becomes more pronounced when the cells experience rapid changes in their environment, and such changes can be easily caused by injury and inflammation, resulting in pain perception or distortion of pain perception (eg, hyperalgesia). Therefore, in this regard, the field of pain is at an advantage to study the epigenetic pathways. More importantly, understanding pain from an epigenetics point of view would provide a new paradigm for developing drugs or strategies for pain management. In this review, we introduce basic concepts of epigenetics, including chromatin dynamics, histone modifications, DNA methylation, and RNA-induced gene silencing. In addition, we provide evidence from published studies suggesting wide implication of different epigenetic pathways within pain pathways. This article provides a brief overview of epigenetic pathways for gene regulation and highlights their involvement in pain. Our goal is to expose the readers to these concepts so that pain-related phenotypes can be investigated from the epigenetic point of view.


Composition-driven surface domain structuring mediated by sphingolipids and membrane-active proteins. Above the nano- but under the micro-scale: mesoscopic biochemical/structural cross-talk in biomembranes.
Biomembranes contain a wide variety of lipids and proteins within an essentially two-dimensional structure. The coexistence of such a large number of molecular species causes local tensions that frequently relax into a phase or compositional immiscibility along the lateral and transverse planes of the interface. As a consequence, a substantial microheterogeneity of the surface topography develops and that depends not only on the lipid-protein composition, but also on the lateral and transverse tensions generated as a consequence of molecular interactions. The presence of proteins, and immiscibility among lipids, constitute major perturbing factors for the membrane sculpturing both in terms of its surface topography and dynamics. In this work, we will summarize some recent evidences for the involvement of membrane-associated, both extrinsic and amphitropic, proteins as well as membrane-active phosphohydrolytic enzymes and sphingolipids in driving lateral segregation of phase domains thus determining long-range surface topography.


A novel microfluidic system to separate sperm using spermatozoa inherent motion and inertial effect.
Sperm separation is an essential part of in vitro fertilization (IVF) process. In conventional procedures, the semen sample is purified from immotile and round cells using centrifugation, which may damage sperm DNA. This study aimed to design a novel microchip to separate the progressively motile spermatozoa using a passive method instead of centrifugation. This microchip is a novel, non-invasive, and two-stage device for auto-selecting the sperm used for IVF. The first stage was designed based on lateral differentiation and rapid divergence to separate the pathways of sperm and round cells. The second stage separates high-quality sperm based on their inherent motion. Before experimenting with fresh human semen samples, preliminary tests were performed using standard particles. The results showed that at the optimized flow rate for separation (1.7 ml/h), the concentration of progressively motile spermatozoa at outlet was significantly increased compared with the initial sample.


How much is that EHR in the window? How to build a cost model that gives the full picture.
Every project decision affects your costs. It comes down to which costs are associated with each element of the project.


Receptor binding characteristics of [3H]NAD-299, a new selective 5-HT1A receptor antagonist.
In vitro receptor binding properties of the novel tritiated 5-hydroxytryptamine1A (5-HT1A) receptor antagonist (R)-3-N,N-dicyclobutylamino-8-fluoro-[6-3H]-3,4-dihydro-2H-1-benzo pyran-5-carboxamide ([3H]NAD-299, generic name robalzotan) were evaluated and compared with those of the agonist 8-hydroxy-2-[2,3-3H]di-n-(propylamino)tetralin ([3H]8-OH-DPAT). [3H]NAD-299 binding displayed a Kd value of 0.17 nM and a Bmax value of 26.7 pmol/g wet weight of rat hippocampus. Same binding affinity (Kd = 0.16 nM) was found to cloned human 5-HT1A receptors. Addition of the nonhydrolyzable GTP analog guanylylimidodiphosphate had no effect on the binding characteristics of [3H]NAD-299, while it significantly decreased both the affinity and density of receptors labeled with [3H]8-OH-DPAT. The rank order of potency of various compounds to inhibit [3H]NAD-299 binding is consistent with the labeling of 5-HT1A receptors. This newly developed high-affinity and selective antagonist radioligand provides a valuable tool for studies of 5-HT1A receptors both in vitro and in vivo.


Are women with thicker cortices in the femoral shaft at higher risk of subtrochanteric/diaphyseal fractures? The study of osteoporotic fractures.
Femoral shaft cortical thickening has been mentioned in reports of atypical subtrochanteric and diaphyseal (S/D) femur fractures, but it is unclear whether thickening precedes fracture or results from a preceding stress fracture and what role bisphosphonates might play in cortical thickening. Our objective was to examine the relationship of cortical thickness to S/D fracture risk as well as establish normal reference values for femoral cortical thickness in a large population-based cohort of older women. Using pelvic radiographs obtained in 1986-1988, we measured femoral shaft cortical thickness 3 cm below the lesser trochanter in women in the Study of Osteoporotic Fractures. We measured this in a random sample and in those with S/D fractures and femoral neck and intertrochanteric fractures. Low-energy S/D fractures were identified from review of radiographic reports obtained between 1986 and 2010. Radiographs to evaluate atypia were not available. Analysis used case-cohort, proportional hazards models. Cortical thickness as a risk factor for low-energy S/D femur fractures as well as femoral neck and intertrochanteric fractures in the Study of Osteoporotic Fractures, adjusting for age and bone mineral density in proportional hazards models. After age adjustment, women with thinner medial cortices were at a higher risk of S/D femur fracture, with a relative hazard of 3.94 (95% confidence interval = 1.23-12.6) in the lowest vs. highest quartile. Similar hazard ratios were seen for femoral neck and intertrochanteric fractures. Medial or total cortical thickness was more strongly related to fracture risk than lateral cortical thickness. In primarily bisphosphonate-naive women, we found no evidence that thick femoral cortices placed women at higher risk for low-energy S/D femur fractures; in fact, the opposite was true. Women with thin cortices were also at a higher risk for femoral neck and intertrochanteric fractures. Whether cortical thickness among bisphosphonate users plays a role in atypical S/D fractures remains to be determined.


Inflammatory Cytokine Profile in Individuals with Inherited Chromosomally Integrated Human Herpesvirus 6.
Acute graft-versus-host-disease (aGVHD) is a major complication following hematopoietic cell transplantations (HCTs). We have shown that HCT recipients in whom either the donor or patient had inherited chromosomally integrated human herpesvirus 6 (iciHHV-6) have a higher incidence of developing more severe aGVHD. Previous studies established that increased proinflammatory cytokines are associated with increased risk for aGVHD and nonrelapse mortality post-HCT. We hypothesized that HCT recipients with donor or recipient iciHHV-6 (iciHHV-6pos HCT cases) will have higher cytokine levels compared with HCT recipients without iciHHV-6 (iciHHV-6neg HCT controls). We identified 64 iciHHV-6pos HCT cases with plasma from days 7, 14, and/or 21 post-HCT and before aGVHD onset in patients who developed aGVHD. We identified 64 iciHHV-6neg HCT controls matched for aGVHD risk factors. We also identified 28 donors with iciHHV-6 and 56 matched donors without iciHHV-6. We measured plasma cytokine concentrations for IL-6, suppression of tumorigenicity 2, T cell immunoglobulin and mucin-domain containing 3, TNFα, soluble TNF receptor 1 (TNFRp55), and C-reactive protein (CRP). We used Mann-Whitney tests and repeated-measures models to compare cytokine levels. iciHHV-6pos HCT cases had higher CRP levels on day 7 and day 21 and higher TNFRp55 levels on day 14 and day 21 compared with iciHHV-6neg HCT controls. These findings were recapitulated in a repeated-measures model. The differences were most evident among patients who subsequently developed aGVHD grades 2 to 4. Additionally, iciHHV-6pos HCT cases had earlier-onset aGVHD (median, 20 versus 27 days post-HCT; P = .02). There were no differences in cytokine levels among healthy donors with or without iciHHV-6. This study demonstrates that HCT recipients with iciHHV-6 have higher proinflammatory cytokines that may be associated with increased risk for aGVHD.


Deep generative modeling for protein design.
Deep learning approaches have produced substantial breakthroughs in fields such as image classification and natural language processing and are making rapid inroads in the area of protein design. Many generative models of proteins have been developed that encompass all known protein sequences, model specific protein families, or extrapolate the dynamics of individual proteins. Those generative models can learn protein representations that are often more informative of protein structure and function than hand-engineered features. Furthermore, they can be used to quickly propose millions of novel proteins that resemble the native counterparts in terms of expression level, stability, or other attributes. The protein design process can further be guided by discriminative oracles to select candidates with the highest probability of having the desired properties. In this review, we discuss five classes of generative models that have been most successful at modeling proteins and provide a framework for model guided protein design.


Breast Cancer Subtype Variation by Race and Ethnicity in a Diverse Population in British Columbia.
Breast cancer subtypes occur differentially across different racial and ethnic groups. However, their distribution within a multicultural population is unknown. Patients with invasive breast cancer newly diagnosed in 2006 and referred to the British Columbia Cancer Agency were identified from the Breast Cancer Outcomes Unit database. Race/ethnicity data were abstracted from a patient-completed health assessment questionnaire completed at the initial consultation, and grouped as white, East Asian, Aboriginal, South Asian, Southeast Asian, and other. Breast cancer subtypes were created using available data on estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER2) status. A total of 1829 women had complete data. Of these women, 73% were white, 8% were East Asian, 4% Aboriginal, 3% South Asian, 3% Southeast Asian, and 3% other. The median age at diagnosis was 60 years; the youngest were Southeast Asian (51 years) and the oldest were white (60 years; P < .001). The overall ER positivity rate was 81%, highest in East Asian women (89%) and lowest in South Asian women (73%). The HER2 positivity rate was 16% for all patients and was highest in the South Asian (28%), Southeast Asian (28%), and Aboriginal (24%) women and lowest in the white women (14%; P < .001). Triple-negative (ER-, PR-, and HER2-negative) breast cancer was uncommon in East Asian women (5%) but more common in South Asian women (19%; P < .001). The 5-year breast cancer-specific survival was 90% (95% confidence interval, 89%-92%), with no significant difference among the racial/ethnic groups (P = .136). Breast cancer subtypes varied by race/ethnicity in our cross-sectional cohort of a multicultural population, suggesting that race/ethnicity plays a significant role in the biology of invasive breast cancer.


Monoclonal antibodies that recognize antigens in human osteosarcoma cells and normal fetal osteoblasts.
Four monoclonal antibodies (HOB-1-4) that react against human osteosarcoma cells (Saos-2) and human fetal osteoblasts in situ were developed by immunizing mice with Saos-2 cells. HOB-1 (IgG1, k) and HOB-2 (IgM, k) stain cytoplasmic antigens in Saos-2 cells, human authentic osteoblasts and occasional cells in liver, spleen and, in the instance of HOB-1, kidney and adrenal gland. On Western blotting of Saos-2 cell lysates, HOB-1 recognizes a single protein species of M(r) 59,000, while HOB-2 reacts with a different species of M(r) 57,000. HOB-3 (IgG1, k) stains a cell membrane associated antigen in Saos-2 cells and osteoblasts in situ. The reaction pattern of this antibody is virtually identical to that seen in alkaline phosphatase (ALP)-positive cells in all organs examined immunohistochemically, except for intestinal epithelium. Both immunoblotting and immunoprecipitation analyses confirm that the antigen detected by HOB-3 is ALP. HOB-4 (IgG1, k) reacts weakly against Saos-2 cells cultured under standard conditions, but strongly following the exposure of the cells to the secretory inhibitor monensin. On frozen section screening, this antibody reacts preferentially with the collagenous extracellular matrix not only of the periosteum but of other tissues and organs as well. The precise identity of the HOB-4 antigen remains to be established. We suggest that these four monoclonal reagents will be useful adjuncts in characterizing the osteoblastic phenotype.


Characterization of free and tightly bound lipoprotein in intima by thin layer isoelectric focusing.
Thin layer isoelectric focusing followed by second dimension quantitative immunoelectrophoresis has been used to characterize the free and tightly bound lipoprotein (lp) fractions in human aortic intima. In 3.3% acrylamide gels plasma low density lipoprotein (LDL) focuses rapidly, but very low density lipoprotein (VLDL) fails to enter the gell and remains at the origin (point of application). Samples of normal intima and lesions were placed directly on the gel and focused for 10--12 h; in normal intima and gelatinous lesions which had not accumulated cholesterol 60--70% of the free LP focused with plasma LDL, but with increasing accumulation of cholesterol the proportion focusing with LDL decreased and the component remaining at the origin increased. In 9 of the 39 samples examined one or more components with pI acid to LDL were present, and in one sample the whole peak focused at a more acid pI. In 4 of 5 lipid-rich centres of gelatinous plaques all the free LP remained at the origin, and the bound LP released by incubation with plasmin remained at the origin in all samples in 3.3% gels although it would focus in 2.6% gels. With antiserum to apo-C this large LP did not behave like VLDL, and it appears to be an aggregated LDL. An increased proportion of aggregated LDL was associated with (a) accumulation of cholesterol, (b) topographical location towards the centre of plaques, and (c) partial destruction of endogenous LP during incubation of fresh samples of tissue.


Protons are one of the limiting factors in determining sensitivity of nano surface-assisted (+)-mode LDI MS analyses.
A proton source employing a nanostructured gold surface for use in (+)-mode laser desorption ionization mass spectrometry (LDI-MS) was evaluated. Analysis of perdeuterated polyaromatic hydrocarbon compound dissolved in regular toluene, perdeuterated toluene, and deuterated methanol all showed that protonated ions were generated irregardless of solvent system. Therefore, it was concluded that residual water on the surface of the LDI plate was the major source of protons. The fact that residual water remaining after vacuum drying was the source of protons suggests that protons may be the limiting reagent in the LDI process and that overall ionization efficiency can be improved by incorporating an additional proton source. When extra proton sources, such as thiolate compounds and/or citric acid, were added to a nanostructured gold surface, the protonated signal abundance increased. These data show that protons are one of the limiting components in (+)-mode LDI MS analyses employing nanostructured gold surfaces. Therefore, it has been suggested that additional efforts are required to identify compounds that can act as proton donors without generating peaks that interfere with mass spectral interpretation.


High levels of microplastics in commercial salt and industrial salterns in Sri Lanka.
This study provides the first analysis and quantification of MPs in salt products in Sri Lanka. Commercial table salt brands, rock salt, lab-grade NaCl and raw salt from three salterns were analysed using microscopy and Fourier transform infrared spectroscopy. All salt samples were contaminated with MPs: in commercial salts products it ranged from 11 to 193 items/kg, rock salts had 64 items/kg and lab grade NaCl had 253 ± 8.9 items/kg. The MP levels in salterns varied significantly: Hambantota 3345.7 ± 311.4 items/kg, Puttalam 272.3 ± 10.6 items/kg, and Elephant Pass 36.3 ± 4.5 items/kg. Predominantly, MPs were presented as fibres, followed by fragments. Of the 23 polymer types identified; low-density polyethylene (LDPE; 17%), resin dispersion (15%) and high-density polyethylene (HDPE; 12%) were notable. This study provides the first comparison of MPs in raw salt and commercial salt. This information is important to trace the pollutant sources and then to take steps to eliminate MPs in food products consumed.


[Evaluation of survival in a sample of patients treated with radical mastectomy for mammary carcinoma].
Survival and its modalities were studied in 40 patients subjected to Halsted's operation for breast cancer between 1960 and 1972. The following parameters were considered: age at time of surgery, onset of tumour before or after menopause, age of commencement of menopause, married or single, number of pregnancies, breast feeding, regularity of menstrual cycle, prior breast disease, time elapsed between first symptoms and mastectomy, side affected, quadrant affected, involvement of ipsilateral lymph nodes, histological type, complementary X-ray treatment, ovariectomy, and medical hormone therapy.


Draft Genome Sequence of Paenibacillus sonchi IIRRBNF1, a Nitrogen-Fixing and Plant Growth-Promoting Bacterium Isolated from Rice Rhizosphere.
Paenibacillus sonchi IIRRBNF1 is a rice-rhizospheric, endospore-forming, Gram-positive, plant growth-promoting rhizobacterium. Here, we report the draft genome sequence of Paenibacillus sonchi IIRRBNF1, which consists of an∼7.3-Mb (7,323,556-bp) genome with 6,271 coding sequences (CDSs), 13 rRNAs, and 67 tRNAs. The genome reveals the presence of a nitrogen-fixing gene cluster and genes associated with multiple plant growth-promoting traits.


Contamination of beef carcasses by psychrotrophic Pseudomonas and Enterobacteriaceae at different stages along the processing line.
The extent of the contamination of beef carcasses with psychrotrophic Pseudomonas spp. and Enterobacteriaceae during slaughter, chilling and cutting was estimated by introducing a new analytical procedure; the contamination index. Comparisons were made between the initial viable counts and the contamination index. The contamination index was calculated as the sum of the bacterial counts obtained during aerobic cold storage of excised meat samples. The presence and composition of spoilage bacteria in the slaughter environment and on the carcasses was also determined at one plant. Rapid chilling was identified as a critical processing step by the contamination index. In addition to this, the dehiding and the chilling in cold storage rooms were implicated as critical operations, with respect to aerosol contamination and surface cross-contamination. Comparison of the composition of spoilage bacteria in the slaughter environment and the bacteria proliferating on the carcass surface samples taken at the corresponding steps showed similar distributions of the identified Pseudomonas spp. In five surveys at two plants, the contamination of beef carcasses along the processing line was estimated. Statistically significant variations between different processing steps were more pronounced for the contamination index than for the conventional counts. It was concluded that the contamination index could be used for identifying critical processing steps, with respect to the extent of contamination of carcasses by psychrotrophic spoilage bacteria.


Protective effect of p-coumaric acid against doxorubicin induced toxicity in H9c2 cardiomyoblast cell lines.
Doxorubicin (Dox) has been used for more than four decades to treat cancer, particularly solid tumours and haematological malignancies. However, the administration of this drug is a matter of concern in the clinical community, since Dox therapy is commonly associated with dose-dependent cardiotoxicity. Attempts at alleviating drug generated cardiac damage using naturally occurring compounds with radical scavenging property are a promising area of research. p-Coumaric acid (pCA) is one such compound which has significant antiradical scavenging effect. This study aims to investigate the effect of pre and co-administration of pCA on mitigating or preventing Dox induced cardiotoxicity in vitro using H9c2 cardiomyoblast cell lines. Addition of pCA and Dox were performed for both treatment and control sets on H9c2 cells. Sulphorhodamine B assay was used to study the cytotoxic effect of pCA and Dox. The effect of the drug on cell morphology, cell viability and nuclear damage was studied using AO/EB and DAPI staining. ROS production was studied using DCFH-DA staining. Mitochondrial membrane potential and intracellular calcium levels were assessed by rhodamine 123 and Fura 2AM staining. pCA showed strong ABTS cation radical scavenging activity and FRAP activity in a dose dependent manner. The results showed that Dox has significant cytotoxic effect in a dose dependent manner while pCA, even at higher concentrations did not display any significant cytotoxicity on H9c2 cells. Both pre treatment and co- administration of pCA reduced the drug induced toxic effects on cell morphology and enhanced the number of viable cells in comparison to the Dox treated cells as evident from the AO/EB and DAPI staining images. The Dox induced ROS production was found to be significantly reduced in pCA pre-treated and co-administered cells. Dox induced changes in mitochondrial membrane potential and intracellular calcium levels were remarkably improved following pre and co-treatment of H9c2 cells with pCA. These results clearly suggest that pre-treatment and co-administration of pCA is a promising therapeutic intervention in managing Dox mediated cardiotoxicity.


Lesions rostral and caudal to the trigeminal motor nucleus and their effects on the jaw-opening reflex of the cat.
Brain stem lesions placed immediately rostral to the trigeminal motor nucleus of the cat abolish the visible jaw-opening reflex produced by electrical stimulation of the periodontal ligaments. These lesions also greatly diminish the amplitude of the electromyographic response recorded from the anterior belly of the digastric muscle. This attenuation could not be modified by injections of systemic strychnine (a glycine antagonist). Lesions placed immediately caudal to the trigeminal motor nucleus also abolished the visible reflex and greatly reduced the amplitude of the digastric response, but in this case, the attenuated response was slightly augmented by strychnine. These data suggest that glycinergic inhibitory neurons form part of the rostral pathway that regulate jaw-opening motor neurons.


Randomized Controlled Trial Assessing the Feasibility of Shortened Fasts in Intubated ICU Patients Undergoing Tracheotomy.
American Society of Anesthesiology guidelines recommend preoperative fasts of 6 hours after light snacks and 8 hours after large meals. These guidelines were designed for healthy patients undergoing elective procedures but are often applied to intubated intensive care unit (ICU) patients. ICU patients undergoing routine procedures may be subjected to unnecessary prolonged fasts. This study tests whether shorter fasts allow for better nutrition delivery and patient outcomes without increasing the risk. Randomized blinded controlled trial. Tertiary academic medical center. ICU patients undergoing bedside tracheotomy. Intubated ICU patients who were receiving enteral feeding and for whom bedside tracheotomy was indicated were enrolled prospectively and randomly allocated to 2 parallel preoperative fasting regimens: a 6-hour fast (control) and a 45-minute fast (intervention). Patients were assessed for aspiration, caloric delivery, metabolic markers, and infectious and noninfectious complications. Twenty-four patients were enrolled and randomized. There were no complications related to the procedure. There were no cases of intraoperative aspiration identified. There was a single postoperative pneumonia in the control group. Median (interquartile range) length of fast and caloric delivery were significantly different between the control group and the shortened fast group: 22 hours (18, 34) vs 14 hours (5, 25; P < .001) and 429 kcal (57, 1125) vs 1050 kcal (825, 1410; P = .01), respectively. Shortening preoperative fasts in intubated ICU patients allowed for better caloric delivery in the preoperative period.


Flow cytometry of keratinocytes.
A prerequisite for using flow cytometry (FCM) is the availability of isolated single cells. Procedures for separation and isolation of keratinocytes from animals and man are available, and the resulting single cell suspensions have been subjected to FCM measurements. The major advantage of the method is the accuracy and speed with which a variety of cellular constituents can be quantified. FCM of keratinocytes has, hitherto, been mainly confined to measurements of nuclear DNA for estimation of cell-cycle distributions and for ploidy studies. In mouse epidermis, cell-cycle distributions were estimated from sequentially obtained DNA histograms and evaluated with other cell kinetic measurements, resulting in new information about epidermal cell-cycle progression, not achievable by any of the methods alone. The best way, therefore, to increase our knowledge of keratinocyte proliferation, is the combined use of DNA FCM and other cell kinetic methods. DNA FCM has also been applied to healthy and diseased human epidermis, and may add valuable information to the classification of skin disease in selected cases. It is believed that further progress in the characterization of keratinocyte growth and development will depend on parameters other than DNA alone.


A copper-based catalytic system for carboxylation of terminal alkynes: synthesis of alkyl 2-alkynoates.
An efficient coupling of terminal alkynes and CO(2) in the presence of alkyl halides can be achieved under ambient conditions using a copper/phosphine catalyst system, providing facile access to a variety of functionalised alkyl 2-alkynoates.


Potential for adverse drug-alcohol interactions among retirement community residents.
Medications and alcohol are both used commonly by older people. Thus, the potential for adverse drug-alcohol interactions is very high in this population, but data on actual concurrent use of alcohol and medicines likely to interact with alcohol are lacking. The objectives of this study were to determine the frequency of alcohol and medication use and the potential for specific adverse drug-alcohol interactions in residents of retirement communities. Cross-sectional study using a mailed survey. Three retirement communities in suburban Milwaukee, Wisconsin. All 454 independently living residents of the communities were surveyed. Of these, 311 residents (68%) returned completed questionnaires. Mean age of respondents was 83 +/- 6 years, 100% were white, 77% were female. The questionnaire included alcohol use questions adapted from the Khavari questionnaire and the CAGE questionnaire to screen for alcohol abuse. Respondents were asked to list all prescription and nonprescription medications. Thirty-eight percent of the population reported using both alcohol and a high risk medication. Six percent had seven or more drinks per week and took a high risk medication. High risk drugs commonly used by drinkers were antihypertensives in 50%, aspirin in 27%, nonsteroidal anti-inflammatory drugs in 20%, medication for congestive heart failure in 18%, antacids or H2 blockers in 16%, sedatives in 11%, narcotics in 5%, and warfarin in 5%. Concurrent use of alcohol and medications is common in residents of these retirement communities. Many of the drugs taken by regular drinkers have potential for adverse drug-alcohol interactions even at moderate levels of alcohol use. This represents a strong possibility of adverse outcomes and a need for increased awareness on the part of both the public and physicians of the potential for interactions between drugs and alcohol.


Environmental exposure to bisphenol analogues and unexplained recurrent miscarriage: A case-control study.
The use of bisphenol A (BPA) has been substantially limited since 2010 due to its toxicity to human health. A group of bisphenol analogues that are structurally similar to BPA have been developed as the alternatives and used widely. The reproductive toxicity of these emerging chemicals has caused substantial concerns in recent years. Whether bisphenol analogues affect miscarriage, especially unexplained recurrent miscarriage (URM), remains to be explored. We conducted a hospital-based, case-control study with 1180 URM cases and 571 controls in China from 2014 to 2016. Concentrations of six bisphenol analogues (BPA, BPAF, BPAP, BPB, BPP and BPS) were measured in the urine samples collected at median intervals of 7.6 months after last miscarriage (interquartile ranges: 4.8, 14.7 months). Multiple logistic regression, Bayesian kernel machine regression (BKMR) and quantile g-computation (q-gcomp) were used to assess the relationship of bisphenol analogues with URM risk. We observed significantly higher levels of all urinary bisphenols in the cases than the controls. After controlling for potential confounders, bisphenol analogues were significantly associated with increased odds of URM in varying degrees. A dose-response pattern was observed for the associations of BPAF, BPAP and BPB quartiles with URM. The mixed exposure of six bisphenol analogues was positively associated with the risk of URM (adjusted odds ratio (aOR) = 1.25; 1.11-1.42), which was mainly driven by BPAP (60.1%), BPAF (25.1%) and BPA (14.8%). After age stratification, the risks tended to be higher in women aged 30 years or older, compared to women <30 years. Our large case-control study indicates that environmental exposure to bisphenol analogues is associated with an increased risk of URM. Older women may be more vulnerable to the insult.


Reconstructing the genotype-to-fitness map for the bacterial chemotaxis network and its emergent behavioural phenotypes.
The signal-transduction network responsible for chemotaxis in Escherichia coli has been characterised in extraordinary detail. Yet, relatively little is known about eco-evolutionary aspects of chemotaxis, such as how the network has been shaped by selection and to what extent natural populations may fine-tune their chemotactic behaviour to the ecological conditions. To address these questions, we here develop an evolutionary-systems-biology model of the chemotaxis network of E. coli, which we apply to estimate the resource accumulation rate (here used as a proxy for fitness) of wildtype and a large number of potential mutant genotypes. Mutant genotypes differ from the wildtype in the concentrations of one or more constituent proteins of the chemotaxis signalling network or in one or more of its kinetic parameters. To guarantee model consistency across the genotype space, we explicitly incorporated biochemical constraints that underly observed phenotypic trade-offs. The model was validated by reconstructing the phenotypic properties of several known mutant genotypes. We also characterised differences in the fitness distribution between genotypes, and reconstructed adaptive walks in genotype space for populations exposed to different environmental conditions. We found that the local fitness landscape is rugged, due to non-additive interactions between mutations. When selection has a consistent direction, just a few adaptive mutations are required to reach a local peak, and different local peaks can be reached by adaptive walks starting from the same initial genotype. However, when the direction of selection is fluctuating, evolutionary paths are much longer and genotype space is explored further. Longer adaptive walks were also observed when evolution was started from a low-fitness genotype such as a CheZ knockout mutant. In line with empirical observations, the initial ΔcheZ mutant did not respond to a step-down stimulus, but a dynamic response similar to the wildtype was recovered following the fixation of compensatory mutations.


Epsilon Aminocaproic Acid Pretreatment Provides Neuroprotection Following Surgically Induced Brain Injury in a Rat Model.
Neurosurgical procedures can damage viable brain tissue unintentionally by a wide range of mechanisms. This surgically induced brain injury (SBI) can be a result of direct incision, electrocauterization, or tissue retraction. Plasmin, a serine protease that dissolves fibrin blood clots, has been shown to enhance cerebral edema and hemorrhage accumulation in the brain through disruption of the blood brain barrier. Epsilon aminocaproic acid (EAA), a recognized antifibrinolytic lysine analogue, can reduce the levels of active plasmin and, in doing so, potentially can preserve the neurovascular unit of the brain. We investigated the role of EAA as a pretreatment neuroprotective modality in a SBI rat model, hypothesizing that EAA therapy would protect brain tissue integrity, translating into preserved neurobehavioral function. Male Sprague-Dawley rats were randomly assigned to one of four groups: sham (n = 7), SBI (n = 7), SBI with low-dose EAA, 150 mg/kg (n = 7), and SBI with high-dose EAA, 450 mg/kg (n = 7). SBI was induced by partial right frontal lobe resection through a frontal craniotomy. Postoperative assessment at 24 h included neurobehavioral testing and measurement of brain water content. Results at 24 h showed both low- and high-dose EAA reduced brain water content and improved neurobehavioral function compared with the SBI groups. This suggests that EAA may be a useful pretherapeutic modality for SBI. Further studies are needed to clarify optimal therapeutic dosing and to identify mechanisms of neuroprotection in rat SBI models.


Utilization of evergreen and decidous oaks by the Californian oak moth Phryganidia californica.
The deciduous oak Quercus lobata was a better quality food source than the evergreen oak Q. agrifolia for the California oak moth Phryganidia californica: it supported higher growth rates, produced higher fecundity and survivorship, and higher efficiency of conversion of ingested food (ECI), efficiency of conversion of digested food (ECD) and efficiency of nitrogen utilization (NUE). Total polyphenols and astringency were similar in both oak speices, whereas water content was lower and leaf specific weight higher in Q agrifolia than in Q lobata. Nitrogen was higher in new leaves of Q agrifolia but higher in mature leaves of Q. lobata. Contrary to most other studies, total polyphenols and astringency were higher in new than in mature leaves. These results confirmed my hypothesis that the differences in leaf quality associated with the evergreen or deciduous life form would translate into food of differing quality for herbivores.


Measurement of immature platelet fraction is useful in the differential diagnosis of MYH9 disorders.
Although the presence of large and giant platelets is important in screening for MYH9 disorders, platelet morphology evaluation is dependent on operator subjectivity. Immature platelet fraction (IPF%) is widely used in clinical practice because of its rapidity and reproducibility; however, IPF% has been rarely analyzed in MYH9 disorders. Therefore, our study aimed to clarify the usefulness of IPF% in the differential diagnosis of MYH9 disorders. We assessed 24 patients with MYH9 disorders, 10 with chronic immune thrombocytopenia (cITP), 14 with myelodysplastic syndromes (MDS) with thrombocytopenia (<100 × 109 /L), and 20 healthy volunteers. Platelet-related data, including IPF% and platelet morphology (diameter, surface area, and staining), were retrospectively analyzed. Median IPF% in MYH9 disorders, 48.7%, was significantly higher than in all other groups (cITP: 13.4%, MDS: 9.4%, controls: 2.6%). IPF% in MYH9 disorders was significantly negatively correlated with platelet count and significantly positively correlated with the diameter and surface area of platelets, but a correlation was not found between IPF% and platelet staining. The area under the curve of IPF% for the differential diagnosis of MYH9 disorders was 0.987 (95% CI: 0.969-1.000), with a sensitivity of 95.8% and specificity of 93.2% when the cutoff value of IPF% was 24.3%. Our study strongly suggests that IPF% is useful in the differential diagnosis between MYH9 disorders and other types of thrombocytopenia.


On the Statistical Testing Methods for Single Laboratory Validation of Qualitative Microbiological Assays with an Unpaired Design.
There exists several statistical methods for detecting a difference of detection rates between alternative and reference qualitative microbiological assays in a single laboratory validation study with an unpaired design. We compared performance of eight methods including Fisher's exact test, unequal variance two-sample t-test, Wilcoxon rank-sum test, z-test, and methods based on Wilson confidence intervals, complementary log-log regression, Firth's logistic regression, and ordinary logistic regression. We first compared the minimum detectable difference in the proportion of detections between the alternative and reference methods among these statistical methods for a varied number of test portions. We then compared power and size of test of these methods using simulated data. Firth's logistic regression and the unequal variance two-sample t-test had the lowest minimum detectable difference and highest power. None of these statistical methods had an estimated size of test always within a 95% confidence interval of the nominal value 0.05 with small numbers of test portions (n = 12, 20, 30). Fisher's exact test, the Wilcoxon rank-sum test, and the z-test were conservative even with a moderately large number of test portions (n = 40), while Firth's logistic regression and the unequal variance two-sample t-test had a size of test closer to 0.05 than other methods. Firth's logistic regression and the unequal variance two-sample t-test are better choices than other competing methods. We recommend the unequal variance two-sample t-test over Firth's logistic regression because the unequal variance two-sample t-test is better known and easier to use. We provide an example using real data.


Short-Term Effects of Strengthening Exercises of the Lower Limb Rehabilitation Protocol on Pain, Stiffness, Physical Function, and Body Mass Index among Knee Osteoarthritis Participants Who Were Overweight or Obese: A Clinical Trial.
Osteoarthritis (OA) of the knee is defined as a progressive disease of the synovial joints and is characterized by wear and tear of the cartilage and underlying bone. This study aimed to determine the short-term effects of the lower limb rehabilitation protocol (LLRP) on pain, stiffness, physical function, and body mass index (BMI) among knee OA participants who were overweight or obese. Methodology. A single-blinded randomized controlled trial of one-month duration was conducted at Rehmatul-Lil-Alameen Postgraduate Institute, Lahore, Pakistan. Fifty overweight or obese participants with knee OA were randomly divided into two groups by a computer-generated number. Participants in the rehabilitation protocol group (RPG) were provided with leaflets explaining the strengthening exercises of the LLRP and instruction of daily care (IDC), while the participants in the control group (CG) were provided with leaflets explaining the IDC only for a duration of four weeks. The primary outcome measures were the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) scores for pain, stiffness, and physical function. The secondary outcome measures were BMI, exercise adherence, and patients' satisfaction assessed by using the numeric rating scale ranging from 0 to 10. The paired-sample t-test was used to analyze the differences within groups from baseline to posttest evaluations. The analysis of variance 2 × 2 factor was used to analyze the differences in BMI, knee pain, stiffness, and physical function between the groups. Participants in the RPG and CG reported a statistically significant reduction in knee pain and stiffness (p ≤ 0.05) within the group. The reduction in the scores of knee pain was higher in participants in the RPG than that in participants in the CG (p=0.001). Additionally, participants in the RPG reported greater satisfaction (p=0.001) and higher self-reported exercise adherence (p=0.010) and coordinator-reported exercise adherence (p=0.046) than the participants in the CG. Short-term effects of the LLRP appear to reduce knee pain and stiffness only, but not physical function and BMI.


Comparison of metaproterenol, isoetharine and salbutamol in the relief of methacholine-induced bronchospasm in dogs.
We evaluated cardiovascular effects and effectiveness of isoetharine, metaproterenol and salbutamol, when administered intratracheally to relieve methacholine-induced bronchospasm in dogs anaesthetized with 50 per cent nitrous oxide, oxygen, halothane and mechanically ventilated. Methacholine 2 micrograms X kg-1 X hour-1 was administrated first followed by halothane (1 MAC) for 30 minutes (control), then metaproterenol, isoetharine or salbutamol. Metaproterenol (15 mg) significantly decreased transpulmonary pressure to 20.1 +/- 0.5 (SE) from 22.5 +/- 1.15 cmH2O (p less than 0.025) after three min and to 15 +/- 0.5 cmH2O (p less than 0.005) after 90 min. Isoetharine (2.5 mg) decreased transpulmonary pressure after five min to 22.1 +/- 1 from 24.5 +/- 1.5 cmH2O (p less than 0.05), and to 21.75 +/- 0.55 mmH2O after 90 min. Salbutamol 25 micrograms X kg-1 decreased transpulmonary pressure to 20.7 +/- 0.75 from 24.25 +/- 1.28 after three min and to 16 +/- .5 after 90 min. The peak effects on airway pressure occurred at 15 min for metaproterenol, 25 min for salbutamol and 20 min for isoetharine. Pulmonary vascular resistance was not significantly changed during halothane anaesthesia alone but decreased significantly after metaproterenol and isoetharine infusion. Heart rate increased ten per cent after metaproterenol, three per cent after isoetharine, and five per cent after salbutamol. No arrhythmias occurred in any group. Cardiac output increased significantly to 3.25 +/- 0.2 from 1.5 +/- 0.17 L X min-1 (p less than 0.025) after metaproterenol to 3.2 +/- .025 from 1.45 +/- .009 after salbutamol and was unchanged after isoetharine. Metaproterenol and salbutamol in the presence of 1 MAC halothane anaesthesia relieved methacholine-induced bronchospasm more rapidly than did isoetharine. The onset of effect was 3 +/- 0.05 min for metaproterenol and salbutamol and 5 +/- 0.01 min for isoetharine. The effect lasted 210 +/- 10.5 min for metaproterenol, 170 +/- 12.5 min for salbutamol and 90 +/- 4.75 min for isoetharine.


[New developments in laser therapy].
Based on the theory of stimulated emission of radiation that was proposed by Albert Einstein in 1916, the first lasers were developed in the 1960s. The first clinical use of laser technology in a German university took place in 1978 in the Department of Dermatology of the Ludwig-Maximilian-University in Munich under the guidance of the former director, Prof. Dr. med. Dr. h.c. mult. Otto Braun-Falco. In the following years, laser technology developed rapidly. Today laser technology is a widely used interdisciplinary therapeutic procedure that has deep clinical and scientific roots in dermatology. There are many conditions in both classic and aesthetic dermatology that are routinely - and sometimes exclusively - treated with lasers. Here we review recent developments in laser medicine. There seems to be a trend to combination procedures. To enhance efficacy, different laser systems are together or lasers are combined with specific topical medications.


Residual Bacteriome after Chemomechanical Preparation of Root Canals in Primary and Secondary Infections.
Secondary infections may be linked to the presence of residual microorganisms within dental root canals. The purpose of this study was to investigate the bacterial composition of primary and secondary root canal infections before and after chemomechanical treatment. Samples were collected before chemomechanical preparation (S1) and before obturation (S2) from 19 subjects (10 primary and 9 secondary infections). DNA was extracted, and the V3/V4 region of the 16S ribosomal RNA gene was amplified using the 347 F/803R primers and paired-end sequenced using the MiSeq (Illumina, San Diego, CA) instrument. Sequencing analysis yielded partial 16S ribosomal RNA gene sequences that were taxonomically classified into 10 phyla and 143 genera. The most prevalent phyla in the S1 and S2 samples were Firmicutes and Bacteroides; however, when comparing between sample groups, Proteobacteria seem to have been enriched in secondary infections. The dominant genera in the primary S1 samples were Bacillus, Streptococcus, and Prevotella, whereas Bacillus, Streptococcus, and Selenomonas dominated the secondary infection S1 samples. Bacillus and Marinilactibacillus were the most dominant genera in the primary and secondary S2 samples. The mean number of operational taxonomic units per sample was 32,656 (±12,124 SD) and 37,113 (±16,994 SD) in the S1 and S2 samples, respectively. Alpha and beta diversities presented the same pattern within samples from both groups. Great interindividual variations in the bacterial composition of the root canal biofilms were observed. There was no difference in the bacterial composition before and after treatment, although some genera survived and seem to be part of a residual microbiome. Our findings revealed a high diversity of the bacterial communities present in root canal infections after chemomechanical treatment, although the majority of the taxa detected were in low abundance.


Complete Genome Sequence of Hydrogen Peroxide-Producing Limosilactobacillus fermentum DM072 Isolated from the Human Oral Cavity.
The complete genome of hydrogen peroxide (H2O2)-producing Limosilactobacillus fermentum strain DM072, isolated from the oral cavity of healthy volunteers in South Korea, was sequenced by long-read sequencing and was subsequently corroborated by short-read sequencing. The genome comprises one circular chromosome and one plasmid and lacks antimicrobial resistance genes.


Alcoholics' beliefs about abuse: causes, effects and treatment.
The beliefs that problem drinkers hold about the causes of abuse, effects of alcohol, and methods of treatment were investigated. The drinkers' beliefs about their own abuse as well as their beliefs about other abusers were both studied. It was found that own abuse was mostly explained by internal factors (drinking to reduce distress or to achieve an elated mood) while external factors (e.g. unemployment) were more often referred to as explanations of the abuse by other persons. Alcohol abuse as an illness was a frequently endorsed concept, while explanations in terms of social or macro level factors were rarely accepted. Females tended to accept more explanations of abuse than males. Many of the problem drinkers tended to see very few explanations for their own abuse and to be quite skeptical about treatment. Factor analysis resulted in three factors: acceptance of reasons for abuse, perceived dependence and experienced control of drinking. Cluster analysis indicated that about 25% of the problem drinkers had a high level of acceptance, perceived dependence and control. These problem drinkers may be a prime target group for treatment.


Treatment of dysfunctional uterine bleeding with danazol.
Twenty-six women completed a 6-month course of treatment with danazol (at various dosages between 200 and 800 mg daily) for menorrhagia due to dysfunctional uterine bleeding. Objective measurements of menstrual blood loss (MBL) were undertaken in 9 women, while the remainder merely recorded a detailed prospective but subjective menstrual calendar. A very substantial decrease in blood loss was recorded by all women, and the majority of women on 400-800 mg daily developed amenorrhoea by 3 months. Six women experienced episodes of prolonged or frequent bleeding or spotting throughout the 6 months. Three women used 200 mg daily throughout with a mean measured MBL falling from 95.3 ml to less than 14 ml per month after 2 months therapy. This may become a valuable therapy for menorrhagia for women requiring temporary medical management. However, possibilities for long-term therapy may be limited in some women by side-effects and metabolic effects.


A general mechanism for transcriptional synergy by eukaryotic activators.
One of the important regulatory concepts to emerge from studies of eukaryotic gene expression is that RNA polymerase II promoters and their upstream activators are composed of functional modules whose synergistic action regulates the transcriptional activity of a nearby gene. Biochemical analysis of synergy by ZEBRA, a non-acidic activator of the Epstein-Barr virus (EBV) lytic cycle, showed that the synergistic transcriptional effect of promoter sites and activation modules correlates with assembly of the TFIID:TFIIA (DA) complex in DNase I footprinting and gel shift assays. The activator-dependent DA complex differs from a basal DA complex by its ability to bind TFIIB stably in an interaction regulated by TATA-binding protein-associated factors (TAFs). TFIIB enhances the degree of synergism by increasing complex stability. Similar findings were made with the acidic activator GAL4-VP16. Our data suggest a unifying mechanism for gene activation and synergy by acidic and non-acidic activators, and indicate that synergy is manifested at the earliest stage of preinitiation complex assembly.


A novel RealTime HIV-1 Qualitative assay for the detection of HIV-1 nucleic acids in dried blood spots and plasma.
Abbott RealTime HIV-1 Qualitative is an in vitro real-time PCR assay for detecting HIV-1 nucleic acids in human plasma and dried blood spots (DBS). The assay was designed to be used in diagnosis of HIV-1 infections in pediatric and adult patients, with an emphasis on the applicability in resource-limited settings. Use of DBS facilitates specimen collection from remote areas and transportation to testing laboratories. Small sample input requirement facilitates testing of specimens with limited collection volume. The Abbott RealTime HIV-1 Qualitative assay is capable of detecting HIV-1 group M subtypes A-H, group O and group N samples. HIV-1 virus concentrations detected with 95% probability were 80 copies/mL of plasma using the plasma protocol, and 2469 copies/mL of whole blood using the DBS protocol. The assay detected HIV-1 infection in 13 seroconversion panels an average 10.5 days earlier than an HIV-1 antibody test and 4.9 days earlier than a p24 antigen test. For specimens collected from 6 weeks to 18 months old infants born to HIV-1 positive mothers, assay results using both the DBS and plasma protocols agreed well with the Roche Amplicor HIV-1 DNA Test version 1.5 (95.5% agreement for DBS and 97.8% agreement for plasma).


Effects of payments for ecosystem services programs in China on rural household labor allocation and land use: Identifying complex pathways.
Payments for Ecosystem Services (PES) is increasingly used in developing countries to secure the sustainable provision of vital ecosystem services. The largest PES programs in the world are embedded in China's new forest policies, which aim to expand forest cover for soil and water conservation and improve livelihoods of rural people. The objective of this study is to identify the complex pathways of impacts of two PES programs - the Conversion of Cropland to Forest Program (CCFP) and the Ecological Welfare Forest Program (EWFP) - on household livelihood decisions, and to quantify the direct and indirect impacts along the identified pathways. We fulfill this objective by developing an integrated conceptual framework and applying a Partial Least Squares-Structural Equation Model (PLS-SEM), based on household survey data from Anhui, China. Labor allocation (for on-farm work, local paid work, local business, and out-migration) and land use decisions (i.e., rent in, maintain, rent out, or abandon cropland) for participating households are key to understand PES program effects on livelihoods. Results show that the PES programs have only small direct effects but significant indirect effects via the mediating factor of capital assets. Moreover, group heterogeneity analysis shows that lower-income households do not benefit any more than the better-off households from the PES, while households with medium wealth increase dependence on agriculture. In addition, household demographics, individual attributes, and geographic settings differ in their impacts on labor allocation and land use decisions. We conclude that CCFP and EWFP programs would be more efficient in conserving the environment while improving the economic welfare of lower-income households if capital assets were taken into account in the design of compensation schemes.


The use of ultraviolet spectra and chromatographic retention data as an aid to metabolite identification.
Phenoxypropanolamines and their putative 4-hydroxy metabolites have been chromatographed using reversed-phase HPLC and UV spectra recorded using a linear diode array detector. Analysis of the chromatographic data using the functional group contribution approach allowed peaks to be tentatively characterized as hydroxy metabolites. This characterization was further enhanced by the examination of the UV spectra which showed distinct changes following hydroxylation. Used in combination, the two techniques allow discrimination between the possible isomeric hydroxy metabolites. The chromatographic and spectral changes observed with the relatively complex phenoxypropanolamines can be successfully modelled using simple compounds to further facilitate identification.


Modeling population size independent tissue epigenomes by ChIL-seq with single thin sections.
Recent advances in genome-wide technologies have enabled analyses using small cell numbers of even single cells. However, obtaining tissue epigenomes with cell-type resolution from large organs and tissues still remains challenging, especially when the available material is limited. Here, we present a ChIL-based approach for analyzing the diverse cellular dynamics at the tissue level using high-depth epigenomic data. "ChIL for tissues" allows the analysis of a single tissue section and can reproducibly generate epigenomic profiles from several tissue types, based on the distribution of target epigenomic states, tissue morphology, and number of cells. The proposed method enabled the independent evaluation of changes in cell populations and gene activation in cells from regenerating skeletal muscle tissues, using a statistical model of RNA polymerase II distribution on gene loci. Thus, the integrative analyses performed using ChIL can elucidate in vivo cell-type dynamics of tissues.


A nested phosphorus and proton coil array for brain magnetic resonance imaging and spectroscopy.
A dual-nuclei radiofrequency coil array was constructed for phosphorus and proton magnetic resonance imaging and spectroscopy of the human brain at 7T. An eight-channel transceive degenerate birdcage phosphorus module was implemented to provide whole-brain coverage and significant sensitivity improvement over a standard dual-tuned loop coil. A nested eight-channel proton module provided adequate sensitivity for anatomical localization without substantially sacrificing performance on the phosphorus module. The developed array enabled phosphorus spectroscopy, a saturation transfer technique to calculate the global creatine kinase forward reaction rate, and single-metabolite whole-brain imaging with 1.4cm nominal isotropic resolution in 15min (2.3cm actual resolution), while additionally enabling 1mm isotropic proton imaging. This study demonstrates that a multi-channel array can be utilized for phosphorus and proton applications with improved coverage and/or sensitivity over traditional single-channel coils. The efficient multi-channel coil array, time-efficient pulse sequences, and the enhanced signal strength available at ultra-high fields can be combined to allow volumetric assessment of the brain and could provide new insights into the underlying energy metabolism impairment in several neurodegenerative conditions, such as Alzheimer's and Parkinson's diseases, as well as mental disorders such as schizophrenia.


Systems Pharmacology-based strategy to screen new adjuvant for hepatitis B vaccine from Traditional Chinese Medicine Ophiocordyceps sinensis.
Adjuvants are common component for many vaccines but there are still few licensed for human use due to low efficiency or side effects. The present work adopted Systems Pharmacology analysis as a new strategy to screen adjuvants from traditional Chinese medicine. Ophiocordyceps sinensis has been used for many years in China and other Asian countries with many biological properties, but the pharmacological mechanism has not been fully elucidated. First in this study, 190 putative targets for 17 active compounds in Ophiocordyceps sinensis were retrieved and a systems pharmacology-based approach was applied to provide new insights into the pharmacological actions of the drug. Pathway enrichment analysis found that the targets participated in several immunological processes. Based on this, we selected cordycepin as a target compound to serve as an adjuvant of the hepatitis B vaccine because the existing vaccine often fails to induce an effective immune response in many subjects. Animal and cellular experiments finally validated that the new vaccine simultaneously improves the humoral and cellular immunity of BALB/c mice without side effects. All this results demonstrate that cordycepin could work as adjuvant to hepatitis b vaccine and systems-pharmacology analysis could be used as a new method to select adjuvants.


Development of bright red-shifted miRFP704nano using structural analysis of miRFPnano proteins.
We recently converted the GAF domain of NpR3784 cyanobacteriochrome into near-infrared (NIR) fluorescent proteins (FPs). Unlike cyanobacterichrome, which incorporates phycocyanobilin tetrapyrrole, engineered NIR FPs bind biliverdin abundant in mammalian cells, thus being the smallest scaffold for it. Here, we determined the crystal structure of the brightest blue-shifted protein of the series, miRFP670nano3, at 1.8 Å resolution, characterized its chromophore environment and explained the molecular basis of its spectral properties. Using the determined structure, we have rationally designed a red-shifted NIR FP, termed miRFP704nano, with excitation at 680 nm and emission at 704 nm. miRFP704nano exhibits a small size of 17 kDa, enhanced molecular brightness, photostability and pH-stability. miRFP704nano performs well in various protein fusions in live mammalian cells and should become a versatile genetically-encoded NIR probe for multiplexed imaging across spatial scales in different modalities.


Ethics committees in critical care.
Ethics committees have emerged as a way for institutions to evaluate and resolve some of the difficult decisions facing critical care medicine. This article examines their origin, scope, function, and potential problems.


The effects of vaccination with tissue culture-derived viral vaccines on detection of antibodies to equine arteritis virus by enzyme-linked immunosorbent assay (ELISA).
An enzyme-linked immunosorbent assay (ELISA) was developed for the detection of serum antibodies to equine arteritis virus (EAV). Results from this assay produced a good correlation with results from virus neutralisation tests in horses which had not been regularly vaccinated with commercially available mammalian tissue culture-derived viral vaccines. Vaccination of some horses with tissue culture-derived vaccines induced the formation of antibodies to bovine serum. These antibodies reacted with the bovine protein contaminants in the EAV ELISA antigen, producing false-positive results. Non-viral protein contaminants were found to be closely associated with EAV in that they co-purified with the virus during gradient centrifugation.


Contralateral ovarian location between the future ovulatory follicle and extant corpus luteum increases the length of the luteal phase and number of follicular waves in heifers.
Location of the future ovulatory follicle and CL in the same ovary (ipsilateral) or opposite ovaries (contralateral) and number of major follicular waves (two or three) per interovulatory interval (IOI) was studied in 14 heifers. Follicle diameter and a blood sample for progesterone (P4) assay were obtained each day throughout an IOI. Heifers were partitioned into three groups: ipsilateral follicle/CL relationship and two follicular waves (Ipsi-2W, N = 5), contralateral relationship and two follicular waves (Contra-2W, N = 5), and contralateral relationship and three waves (Contra-3W, N = 4). Only one heifer had an ipsilateral relationship and three waves and was not included in the analyses. An unexpected observation was slower growth of the dominant follicle of Wave 1 in the Ipsi-2W group than in the Contra-2W and Contra-3W groups. Increased P4 production in the Contra-3W group compared with the Ipsi-2W and Contra-2W groups was indicated by significantly greater P4 concentration averaged over Days 0 to 20 (Day 0 = ovulation), longer interval from ovulation to the beginning of a decrease in P4 and to the beginning of postluteolysis (P4 <1 ng/mL), and longer IOI. The interval from the beginning of postluteolysis to ovulation was not different among groups, indicating that the prolonged IOI reflected the prolonged luteal phase. An effect of the follicle/CL relationship on length of the IOI was not detected in mares. Results supported the hypothesis that the prolonged luteal phase of the contralateral follicle/CL relationship favors the development of three follicular waves/IOI in heifers.


Clinical and microbiological effects of mechanical instrumentation and local antimicrobials during periodontal supportive therapy in aggressive periodontitis patients: smoker versus non-smoker patients.
To compare the clinical and microbiological effects of ultrasonic mechanical instrumentation (UMI) associated to home-care use of amine fluoride/stannous fluoride (AmF/SnF(2) )-containing mouthrinse and toothpaste in smoker and non-smoker patients affected by generalized aggressive periodontitis (G-AgP) during a recall session of supportive periodontal therapy (SPT). Thirteen smokers and 25 non-smokers G-AgP patients enrolled in an SPT programme received a single session of UMI associated with home-care use of AmF/SnF(2) -containing mouthrinse and toothpaste. Clinical and microbiological parameters were assessed pre-treatment, at 6 and 12 weeks post-treatment. In both groups, UMI plus AmF/SnF(2) -implemented oral hygiene use determined a significant decrease of total bacterial counts, with non-smokers exhibiting a lower count compared with smokers at 12 weeks. No significant differences were observed between smokers and non-smokers in the counts of total pathogens and red complex species at each observation interval. Clinically, a significant reduction of supragingival plaque, gingival inflammation and probing pocket depth was similarly observed in both groups. A combined mechanical/chemical plaque control approach based on UMI and the use of AmF/SnF(2) agents resulted in the reduction of supragingival plaque deposits, gingival inflammation and subgingival periodontal pathogens in G-AgP patients during SPT, with no substantial difference between smokers and non-smokers.


Incidence of Anterior Knee Pain and Functional Difficulty in a Prospectively Recruited Cohort Following Total Knee Arthroplasty With Selective Patellar Resurfacing.
The influence of patellar resurfacing on anterior knee pain, stair climbing, and functional activity outcomes following total knee arthroplasty (TKA) are incompletely understood. This study examined the influence of patellar resurfacing on patient-reported outcome measures (PROMs) relating to anterior knee pain and function. The Knee Injury and Osteoarthritis Outcome Score of Joint Replacement (KOOS, JR.) patient PROMs were collected preoperatively and at 12 months follow-up for 950 TKAs performed over 5 years. Indications for patellar resurfacing included Grade IV patello-femoral (PFJ) changes or mechanical PFJ findings during patellar trialing. Patellar resurfacing was performed on 393 (41%) of the 950 TKAs performed. Multivariable binomial logistic regressions were performed, using KOOS, JR. questions assessing pain during stair climbing, standing upright, and function during rising from sitting as surrogates for anterior knee pain. Independent regression models were performed for each of the targeted KOOS, JR. questions, with adjustment for age at surgery, sex, and baseline pain and function. No association was observed between 12-month postoperative anterior knee pain or function with patellar resurfacing (P = .17, .97). Patients who had moderate or greater preoperative pain on stairs had an increased likelihood of postoperative pain and functional difficulty (odds ratio 2.3, P = .013), while males were 42% less likely to report postoperative anterior knee pain (odds ratio 0.58, P = .002). Selective patellar resurfacing based on PFJ degeneration and mechanical PFJ symptoms produces similar improvement in PROMs for resurfaced and unresurfaced knees.


Targeted α-therapy using astatine (211At)-labeled PSMA1, 5, and 6: a preclinical evaluation as a novel compound.
Targeted α-therapy (TAT) for prostate-specific membrane antigen (PSMA) is a promising treatment for metastatic castration-resistant prostate cancer (CRPC). Astatine is an α-emitter (half-life=7.2 h) that can be produced by a 30-MeV cyclotron. This study evaluated the treatment effect of 211At-labeled PSMA compounds in mouse xenograft models. Tumor xenograft models were established by subcutaneous transplantation of human prostate cancer cells (LNCaP) in NOD/SCID mouse. [211At]PSMA1, [211At]PSMA5, or [211At]PSMA6 was administered to LNCaP xenograft mice to evaluate biodistribution at 3 and 24 h. The treatment effect was evaluated by administering [211At]PSMA1 (0.40 ± 0.07 MBq), [211At]PSMA5 (0.39 ± 0.03 MBq), or saline. Histopathological evaluation was performed for the at-risk organs at 3 and 6 weeks after administration. [211At]PSMA5 resulted in higher tumor retention compared to [211At]PSMA1 and [211At]PSMA6 (30.6 ± 17.8, 12.4 ± 4.8, and 19.1 ± 4.5 %ID/g at 3 h versus 40.7 ± 2.6, 8.7 ± 3.5, and 18.1 ± 2.2%ID/g at 24 h, respectively), whereas kidney excretion was superior in [211At]PSMA1 compared to [211At]PSMA5 and [211At]PSMA6. An excellent treatment effect on tumor growth was observed after [211At]PSMA5 administration. [211At]PSMA1 also showed a substantial treatment effect; however, the tumor size was relatively larger compared to that with [211At]PSMA5. In the histopathological evaluation, regenerated tubules were detected in the kidneys at 3 and 6 weeks after the administration of [211At]PSMA5. TAT using [211At]PSMA5 resulted in excellent tumor growth suppression with minimal side effects in the normal organs. [211At]PSMA5 should be considered a new possible TAT for metastatic CRPC, and translational prospective trials are warranted.