In vitro activity of aminoglycosides against clinical isolates of Acinetobacter baumannii complex and other nonfermentative Gram-negative bacilli causing healthcare-associated bloodstream infections in Taiwan.
Aminoglycosides possess in vitro activity against aerobic and facultative Gram-negative bacilli. However, nationwide surveillance on susceptibility data of Acinetobacter baumannii complex and Pseudomonas aeruginosa to aminoglycosides was limited, and aminoglycoside resistance has emerged in the past decade. We study the in vitro susceptibility of A. baumannii complex and other nonfermentative Gram-negative bacilli (NFGNB) to aminoglycosides. A total of 378 NFGNB blood isolates causing healthcare-associated bloodstream infections during 2008 and 2013 at four medical centers in Taiwan were tested for their susceptibilities to four aminoglycosides using the agar dilution method (gentamicin, amikacin, tobramycin, and isepamicin) and disc diffusion method (isepamicin). A. baumannii was highly resistant to all four aminoglycosides (range of susceptibility, 0-4%), whereas >80% of Acinetobacter nosocomialis and Acinetobacter pittii blood isolates were susceptible to amikacin (susceptibility: 96% and 91%, respectively), tobramycin (susceptibility: 92% and 80%, respectively), and isepamicin (susceptibility: 96% and 80%, respectively). All aminoglycosides except gentamicin possessed good in vitro activity (>94%) against P. aeruginosa. Amikacin has the best in vitro activity against P. aeruginosa (susceptibility, 98%), followed by A. nosocomialis (96%), and A. pittii (91%), whereas tobramycin and isepamicin were less potent against A. pittii (both 80%). Aminoglycoside resistances were prevalent in Stenotrophomonas maltophilia and Burkholderia cepacia complex blood isolates in Taiwan. Genospecies among the A. baumannii complex had heterogeneous susceptibility profiles to aminoglycosides. Aminoglycosides, except gentamicin, remained good in vitro antimicrobial activity against P. aeruginosa. Further in vivo clinical data and continuous resistance monitoring are warranted for clinical practice guidance.


A comparison between two emergency cricothyroidotomy instruments.
As part of a training programme for younger doctors, two commercially available, emergency cricothyroidotomy sets were evaluated. Prior to the surgical procedure, half of the doctors in each group received an audiovisual lesson. In a simulated but realistic emergency situation, involving autopsy material, cricothyroidotomy was performed utilizing either the "Nutrake" or the "Gentofte" system. The audiovisual lesson increased the speed and the success rate of the doctors performing this procedure as well as strengthening their confidence. Both systems allowed positive pressure ventilation. The participating doctors achieved a higher success rate at a faster speed with the "Nutrake" set.


Adolescent social isolation changes social recognition in adult mice.
Rearing in social isolation has profound effects on several aspects of behavior in adult rodents. However, little is known about effects of social stress on social behavior in these animals. In the present study, we examined social recognition in mice of both sexes that were individually housed from 30 days of age until testing at approximately 80 days of age, individually housed from day 30 until day 60, followed by group housing from day 60 until testing at around 80 days of age and in control mice that were group housed throughout experiment. A standard social recognition test was performed with ovariectomized female conspecifics introduced into the home cage of tested mice for 1 min, eight consecutive times with 9 min breaks between tests, and in the ninth test, new, unfamiliar females were introduced. The time spent investigating stimulus mice during each of the nine tests was recorded. Group housed male and female mice showed strong pattern of social learning, whereas mice reared in isolation from day 30 until testing did not show evidence of social recognition. Interestingly, mice reared in isolation from 30 until 60 days of age and then group housed again, also showed reduced ability for social learning in comparison to the controls housed in groups through the entire period. These results therefore show that social isolation has a profound effect on social behavior in mice, and that even isolation for a limited period can produce lasting behavioral deficits.


Microdetermination of the prostaglandin B1 in human plasma by gas chromatography/selected ion monitoring using [18O]prostaglandin B1 as an internal standard.
We devised a simple and effective purification for the microdetermination of prostaglandin B1 (PGB1), a metabolite of PGE1. [18O]PGB1 was synthesized by the repeated base-catalyzed hydrolysis of methyl ester derivatives in H(2)18O, to obtain an internal standard for the gas chromatography/selected ion monitoring (GC/SIM) of PGB1. The methyl ester-methoxime-dimethylisopropylsilyl ether derivative was prepared, then GC/SIM was carried out by monitoring the ion at m/z 448.3 for PGB1 and that at m/z 452.3 for internal standard. A good linear response over the range of 10 pg to 100 ng was demonstrated. We detected PGB1 to a level of about 40 pg/mL in human plasma. This method can be used to determine PGB1 in biological samples.


Endogenous, tissue-resident stem/progenitor cells in gonads and bone marrow express FSHR and respond to FSH via FSHR-3.
Follicle stimulating hormone (FSH) is secreted by the anterior pituitary and acts on the germ cells indirectly through Granulosa cells in ovaries and Sertoli cells in the testes. Extragonadal action of FSH has been reported but is still debated. Adult tissues harbor two populations of stem cells including a reserve population of primitive, small-sized, pluripotent very small embryonic-like stem cells (VSELs) and slightly bigger, tissue-specific progenitors which include ovarian stem cells (OSCs) in ovaries, spermatogonial stem cells (SSCs) in testes, endometrial stem cells (EnSCs) in uterus and hematopoietic stem cells (HSCs) in the bone marrow. Data has accumulated in animal models showing FSHR expression on both VSELs and progenitors in ovaries, testes, uterus and bone marrow and eventually gets lost as the cells differentiate further. FSH exerts a direct action on the stem/progenitor cells via alternatively spliced FSHR-3 rather than the canonical FSHR-1. FSH stimulates VSELs to undergo asymmetrical cell divisions to self-renew and give rise to the progenitors that in turn undergo symmetrical cell divisions and clonal expansions followed by differentiation into specific cell types. Excessive self-renewal of VSELs results in cancer and this explains ubiquitous expression of embryonic markers including nuclear OCT-4 along with FSHR in cancerous tissues. Focus of this review is to compile published data to support this concept. FSHR expression in stem/progenitor cells was confirmed by immuno-fluorescence, Western blotting, in situ hybridization and by quantitative RT-PCR. Two different commercially available antibodies (Abcam, Santacruz) were used to confirm specificity of FSHR expression along with omission of primary antibody and pre-incubation of antibody with immunizing peptide as negative controls. Western blotting allowed detection of alternatively spliced FSHR isoforms. Oligoprobes and primers specific for Fshr-1 and Fshr-3 were used to study these alternately-sliced isoforms by in situ hybridization and their differential expression upon FSH treatment by qRT-PCR. To conclude, stem/progenitor cells in adult tissues express FSHR and directly respond to FSH via FSHR-3. These findings change the field of FSH-FSHR biology, call for paradigm shift, explain FSHR expression on cancer cells in multiple organs and provide straightforward explanations for various existing conundrums including extragonadal expression of FSHR.


Prediction of supraclavicular lymph node metastasis in breast carcinoma.
Supraclavicular lymph node metastasis in breast cancer patients has a poor prognosis, and aggressive local treatment has usually resulted in severe morbidity. The purpose of this study was to select high-risk neck metastasis patients for prophylactic radiotherapy. Between 1990 and 1998, 2658 consecutive invasive breast cancer patients underwent surgery and adjuvant therapy in the hospital. The median age was 47 years (range 22-92). The median follow-up period was 39 months. The following factors were analyzed: age, tumor size, tumor location, histologic type, histologic grade, estrogen and progesterone receptor status, DNA flow cytometry study results, number of positive axillary lymph nodes, use of chemotherapy, radiotherapy, and/or hormonal therapy, and level of involved axillary nodes. Of the 2658 patients, 113 (4.3%) developed supraclavicular lymph node metastasis during this period. Young age (< or =40 years), tumor size >3 cm, high histologic grade, angiolymphatic invasion, negative estrogen receptor status, synthetic phase fraction >4%, >4 positive nodes, and level II or III involved nodes were all significant for predicting neck metastasis in the univariate analysis. Three predictive factors were significant after multivariate analysis: high histologic grade, >4 positive nodes, and axillary level II or III involved nodes. In patients with axillary level I involved nodes and < or =4 positive nodes, the incidence was 4.4%. If axillary level III was involved, the rate of supraclavicular lymph node metastasis was 15.1%. The incidence of supraclavicular lymph node metastasis was higher in the groups with >4 positive nodes and in those with axillary level II or III involved nodes. Selective use of comprehensive radiotherapy for these high-risk patients will achieve good locoregional control.


A new release of MOPAC incorporating the INDO/S semiempirical model with CI excited states.
The semiempirical INDO/S Hamiltonian is incorporated into a new release of MOPAC2016. The MOPAC2016 software package has long been at the forefront of semiempirical quantum chemical methods (SEQMs) for small molecules, proteins, and solids and until this release has included only NDDO-type SEQMs. The new code enables the calculation of excited states using the INDO/S Hamiltonian combined with a configuration interaction (CI) approach using single excitations (CIS), single and double excitations (CISD), or multiple reference determinants (MRCI) where reference determinants are generated using a complete active space (CAS) approach. The capacity to perform excited-state calculations beyond the CIS level makes INDO/CI one of the few low-cost computational methods capable of accurately modeling states with substantial double-excitation character. Solvent corrections to the ground-state and excited-state energies can be computed using the COSMO implicit solvent model, incorporating state-specific corrections to the excited states based on the solvent refractive index. This code produces physically reasonable electronic structures, absorption spectra, and solvatochromic shifts at low computational costs for systems up to hundreds of atoms, and for both organic molecules and metal clusters.


[Interstitial gamma-therapy of patients with cancer of the mucous membrane of the mouth].
In 1984-1986, interstitial gamma-therapy was used in 57 patients for cancer of the oral cavity (28), tongue (16) and lower lip (13). Complete regression was observed in 44 (77.3%) patients. No severe side effects were registered. The treatment proved less effective in cases of large tumor located in proximal areas of the oral cavity.


Effect of liposome-entrapped methotrexate on Ehrlich ascites tumor cells and uptake in primary liver cell tumor.
A therapeutically useful concentration (0,5 mg/ml) of Methotrexate was prepared in negatively charged artificial liposomes (lecithine: cholesterol: dicetylphosphate=5:5:1 molar ratios). Entrapment yield after separation on Sepharose 6B is nearly 100%. In contrast to free Methotrexate the liposome entrapped folic acid antagonist is eliminated only slowly by the kidneys and after intravenous application it is unevenly distributed in the body of mice, the highest concentrations being found in liver and spleen. Daily injections (7.5 mg/kg/day) of entrapped Methotrexate for 5 days into the tail vein of Ehrlich ascites tumor bearing mice reduced both tumor cell count and the production of ascites fluid about fourfold as compared to mice receiving the same dose of Methotrexate in the free form. Six hours after intravenous application of liposome entrapped Methotrexate the tissue concentration in normal liver is ca. 20-fold higher than when the same amount is applied in the free state. On the other hand, only a little uptake of liposome entrapped Methotrexate was detected in the tissue of nitrosamine-induced primary liver tumor when compared to normal liver tissue of rats.


Control of cleavage site selection during mRNA 3' end formation by a yeast hnRNP.
Endonucleolytic cleavage of pre-mRNAs is the first step during eukaryotic mRNA 3' end formation. It has been proposed that cleavage factors CF IA, CF IB and CF II are required for pre-mRNA 3' end cleavage in yeast. CF IB is composed of a single polypeptide, Nab4p/Hrp1p, which is related to the A/B group of metazoan heterogeneous nuclear ribonucleoproteins (hnRNPs) that function as antagonistic regulators of 5' splice site selection. Here, we provide evidence that Nab4p/Hrp1p is not required for pre-mRNA 3' end endonucleolytic cleavage. We show that CF IA and CF II devoid of Nab4p/Hrp1p are sufficient to cleave a variety of RNA substrates but that cleavage occurs at multiple sites. Addition of Nab4p/Hrp1p prevents these alternative cleavages in a concentration-dependent manner, suggesting an essential and conserved role for some hnRNPs in pre-mRNA cleavage site selection.


[An intracutaneous test with gonococcal allergen in married couples with impaired reproductive function].
Comparative study of the diagnostic value of intracutaneous test with Gonococcus allergen, bacterioscopic and bacteriologic methods used in examinations of 200 married couples suffering from infertility have demonstrated a high sensitivity of the intracutaneous test. This test is recommended for wide use in screenings for gonorrhea in examinations of patients with impaired reproductive function.


Zika virus infection induces expression of NRF2 and antioxidant systems in trophoblast cells.
The nuclear factor erythroid 2-related factor 2 (NRF2) is a transcription factor that plays a critical role in the xenobiotic and stress responses. During viral infection, NRF2 can modulate the host metabolism and innate immunity; however, the most common activity of NRF2 in viral diseases is controlling reactive oxygen species (ROS). The Zika virus (ZIKV) is involved in a vertical infection in pregnancy, with reported fetal health consequences. However, the possibility that ZIKV regulates NRF2 expression in placental trophoblasts has not been investigated. In this report, we evaluated the upregulation of NRF2 and antioxidant enzymes in a trophoblast-like cell. These findings could help us understand the antioxidant mechanism underlying the ZIKV infection in the placenta during pregnancy.


Recent advances in ZnO nanostructure as a gas-sensing element for an acetone sensor: a short review.
Air pollution is a severe concern globally as it disturbs the health conditions of living beings and the environment because of the discharge of acetone molecules. Metal oxide semiconductor (MOS) nanomaterials are crucial for developing efficient sensors because of their outstanding chemical and physical properties, empowering the inclusive developments in gas sensor productivity. This review presents the ZnO nanostructure state of the art and notable growth, and their structural, morphological, electronic, optical, and acetone-sensing properties. The key parameters, such as response, gas detection limit, sensitivity, reproducibility, response and recovery time, selectivity, and stability of the acetone sensor, have been discussed. Furthermore, gas-sensing mechanism models based on MOS for acetone sensing are reported and discussed. Finally, future possibilities and challenges for MOS (ZnO)-based gas sensors for acetone detection have also been explored.


[Functional reserves of the sympathetic apparatus of the heart in hypoxia].
The functional reserves of sympathetic systems, i.e. their capacity to stabilize the level of norepinephrine in the enlarged myocardium of rats exposed to chronic hypoxia, increased with their adaptation to hypoxia and decreased with their deadaptation. The level of these reserves depended at large on the rate of norepinephrine biosynthesis. The degree of myocardial hypertrophy of this genesis increased with prolongation of the hypoxic exposure. It is concluded that optimal supply of norepinephrine to the enlarged heart due to increased functional reserves of sympathetic systems is possible only up to a certain level of an increase in the myocardial mass.


Solution structure ensemble of human obesity-associated protein FTO reveals druggable surface pockets at the interface between the N- and C-terminal domain.
The fat mass and obesity-associated FTO protein catalyzes demethylation of the N6-methyladenosine, an epigenetic mark that controls several metabolic pathways by modulating the transcription, translation, and cellular localization of RNA molecules. Since the discovery that its overexpression links to the development of obesity and cancer, FTO was the target of screening campaigns and structure-based drug design efforts. Although several FTO inhibitors were generated, these often lack potency or selectivity. Herein, we investigate the structure and dynamics of human FTO in solution. We show that the structure of the catalytic N-terminal domain is unstable in the absence of the C-terminal domain, which explains why the isolated N-terminal domain is incompetent for catalysis and suggests that the domain interaction represents a target for the development of specific inhibitors. Then, by using NMR relaxation measurements, we show that the interface between the FTO structural domains, the active site, and several peripheral loops undergo conformational dynamics on both the picosecond-nanosecond and microsecond-millisecond timescales. Consistent with this, we found that the backbone amide residual dipolar couplings measured for FTO in phage pf1 are inconsistent with the static crystal structure of the enzyme. Finally, we generated a conformational ensemble for apo FTO that satisfies the solution NMR data by combining the experimental residual dipolar couplings with accelerated molecular dynamics simulations. Altogether, the structural ensemble reported in this work provides an atomic-resolution model of apo FTO and reveals transient surface pockets at the domain interface that represent potential targets for the design of allosteric inhibitors.


What is the impact of the explicit knowledge of sequence regularities on both deterministic and probabilistic serial reaction time task performance?
The aim of this study was to explore the role of prior explicit sequence knowledge by comparing its influence on serial reaction time (SRT) performance with either a deterministic or a probabilistic sequence. The results confirm that, with a deterministic sequence, preliminary explicit learning improves SRT performance. On the other hand, with a probabilistic sequence, the results show no advantage for SRT performance in explicit-learning conditions. In addition, by using the process dissociation procedure (Jacoby, 1991), we show that performance on a subsequent generation task was more sustained by controlled processes for participants in the explicit-learning conditions than for those in the incidental condition. On the whole, these results, showing that the influence of explicit knowledge can be suppressed in certain specific conditions, are consistent with the intervention of both implicit and explicit mechanisms in SRT tasks, and the results also show that their relative influence can be modulated by the particular demands of the task.


Molecular characterisation of the Ancylostoma-secreted protein family from the adult stage of Ancylostoma caninum.
The Ancylostoma-secreted proteins are a family of nematode-specific cysteine-rich secreted proteins belonging to the pathogenesis-related protein superfamily. Previously we reported that third stage infective larvae of Ancylostoma caninum produce two different Ancylostoma-secreted proteins, a single and double-domain Ancylostoma-secreted protein, designated as Ancylostoma-secreted protein-1 and Ancylostoma-secreted protein-2, respectively. Here we report that adult A. caninum hookworms produce and release four additional Ancylostoma-secreted proteins (Ancylostoma-secreted protein-3-6). Using antiserum against adult excretory/secretory products, Ancylostoma-secreted protein cDNAs were isolated from cDNA expression libraries. Immunolocalisation experiments using specific antisera indicated that the single-domain Ac-Ancylostoma-secreted protein-3 is located in the adult pharyngeal and oesophageal glands. Ac-Ancylostoma-secreted protein-4, Ancylostoma-secreted protein-5 and Ancylostoma-secreted protein-6 are composed of two pathogenesis-related protein domains linked in tandem as a heterodimorphic repeat. Ac-Ancylostoma-secreted protein-4 is localised to the cuticular surface of the adult hookworm, whereas Ac-Ancylostoma-secreted protein-5 was found in the intestinal brush border membrane, and Ancylostoma-secreted protein-6 in the cephalic and excretory glands. All of the adult Ancylostoma-secreted proteins were identified in excretory/secretory products of adult hookworms by Western blotting and are presumably released by the parasite. None of the adult Ancylostoma-secreted proteins were detected by immunoblotting in L3 extracts, although mRNAs of Ac-Ancylostoma-secreted protein-3 and Ac-Ancylostoma-secreted protein-4 were present in the larval stage. The functions of the adult Ancylostoma-secreted proteins are unknown, although the secretion of multiple family members by the adult suggests an important role in the establishment or maintenance of the parasitic relationship.


Human insulin receptor monoclonal antibody undergoes high affinity binding to human brain capillaries in vitro and rapid transcytosis through the blood-brain barrier in vivo in the primate.
The ability of monoclonal antibodies against the human insulin receptor to undergo transcytosis through the blood-brain barrier (BBB) was examined in the present studies. Two murine monoclonal antibodies (MAb83-7 and MAb83-14) which bind different epitopes within the alpha-subunit of the human insulin receptor were examined using isolated human brain capillaries, frozen sections of primate brain, and in vivo pharmacokinetic studies in anesthetized Rhesus monkeys. Both antibodies strongly illuminated capillary endothelium in immunocytochemical analysis of frozen sections of brain from Rhesus monkey but not squirrel monkey. Both monoclonal antibodies, in the iodinated forms, bound to human brain microvessels, although the binding and endocytosis of MAb83-14 was approximately 10-fold greater than MAb83-7. The active binding of MAb83-14 to the human insulin receptor was paralleled by a very high rate of transport of this antibody through the BBB in vivo in two anesthetized Rhesus monkeys. The BBB permeability-surface area (PS) product in neocortical gray matter was 5.4 +/- 0.6 microL/min/g, which is severalfold greater than previous estimates of the PS product for receptor-specific monoclonal antibody transport through the BBB. The brain delivery of MAb83-14 to the Rhesus monkey brain was high and 3.8 +/- 0.4% of the injected dose was delivered to 100 g of brain at 3 hours after a single intravenous injection. In contrast, there was no brain uptake of the mouse IgG2a isotype control antibody. These studies demonstrate an unexpected high degree of transcytosis of a monoclonal antibody through the primate BBB in vivo.


Upscaling biological complexity to boost neuronal and oligodendroglia maturation and improve in vitro developmental neurotoxicity (DNT) evaluation.
Human induced pluripotent stem cell (iPSC)-derived neuronal and glial cell models are suitable to assess the effects of environmental chemicals on the developing brain. Such test systems can recapitulate several key neurodevelopmental features, such as neural stem cell formation and differentiation towards different neuronal subtypes and astrocytes, neurite outgrowth, synapse formation and neuronal network formation and function, which are crucial for brain development. While monolayer, two-dimensional (2D) cultures of human iPSC-neuronal or glial derivatives are generally suited for high-throughput testing, they also show some limitations. In particular, differentiation towards myelinating oligodendrocytes can only be achieved after extended periods in differentiation. In recent years, the implementation of three-dimensional (3D) neuronal and glial models obtained from human iPSCs has been shown to compensate for such limitations, enabling robust differentiation towards both neuronal and glial cell populations, myelination and formation of more mature neuronal network activity. Here we compared the differentiation capacity of human iPSC-derived neural stem cells cultured either as 2D monolayer or as 3D neurospheres, and assessed chlorpyrifos (CPF) effects. Data indicate that 3D neurospheres differentiate towards neurons and oligodendroglia more rapidly than 2D cultures; however, the 2D model is more suitable to assess neuronal functionality by analysis of spontaneous electrical activity using multielectrode array. Moreover, 2D and 3D test systems are diversely susceptible to CPF treatment. In conclusion, the selection of the most suitable in vitro test system (either 2D or 3D) should take into account the context of use and intended research goals ('fit for purpose' principle).


Rectal administration of antiepileptic drugs in children.
This article reviews the current literature describing the use of rectally administered antiepileptic drugs. Individual antiepileptic drugs are discussed in regard to efficacy, toxicity, and rate of absorption. Absorption occurs through passive diffusion; therefore, solutions are absorbed most quickly. Paraldehyde, diazepam, secobarbital, and valproic acid are used when rapid effect for termination of prolonged or serial seizures is desired. Valproic acid, lorazepam, carbamazepine, and phenytoin all can be used for maintenance therapy.


Temporary Threshold Shifts in Naïve and Experienced Belugas: Can Dampening of the Effects of Fatiguing Sounds Be Learned?
In belugas (Delphinapterus leucas), substantial (10-15 dB) differences in temporary threshold shifts (TTSs) were observed between the first and subsequent experimental sessions in the same subjects. In the first session (naïve subject state), the TTSs produced by exposure to fatiguing noises were larger than the TTSs produced in subsequent sessions (experienced subject state). After one to two sessions, the TTSs stabilized. The baseline hearing thresholds did not differ between the naïve and experienced states. One possible explanation for this effect is that the animals learned to dampen their hearing during exposure to fatiguing noises and thus mitigate the impact of those noises.


Multivariable optimization of a multiproduct continuous chemicals facility.
Multivariable optimization (MVO) is a powerful nonlinear steady-state flowsheet simulation technique used widely in the chemical process industry to optimize plant performance by increasing plant capacity and/or reducing energy usage. The user supplies the objective function(s), constraints, and variable limits based on operating heuristics, prior experience, and observed process behavior. In this paper we describe how we used MVO in conjunction with improved automation and statistical process monitoring to increase capacity and reduce energy consumption. This project was conducted over a two-year period in a methylamines facility that produces three products: monomethylamine (MMA), dimethylamine (DMA), and trimethylamine (TMA). It led to a 14% improvement in MMA capacity and a 7% improvement in TMA capacity. Energy consumption per pound of main product decreased by approximately 20% for the six-month period (April-September, 2000) when compared to a three-year average (1997-2000). It was accompanied by a 60% improvement in process capability and 31% improvement in product quality. We attribute this improvement to several factors: smoother transition between optimum set points determined by the process optimizer, improved disturbance rejection due to controller installation and tuning, and reduced operator intervention because of controlled operation at or near the optimum set points.


[Evaluation of the quality of recovery and the postoperative health status after elective surgery].
Postoperative recovery is a complex process with physiologic, functional, and psychologic dimensions. Postoperative quality of recovery is considered as a crucial outcome following surgery and anesthesia. The objective of this study was to assess and compare the quality of postoperative recovery and health status before and after surgery, in patients undergoing elective surgery. This observational, prospective study was conducted on patients proposed for elective surgery. Evaluation of postoperative recovery was performed using the Postoperative Quality of Recovery Scale and health status was assessed by applying the EuroQol assessing problems in five dimensions: mobility, personal care, usual activities, pain/discomfort, and anxiety/depression, and the World Health Organization Disability Assessment Schedule 2.0. Poor quality of recovery was defined as recovery in fewer than two domains at postoperative Day 1 in the Postoperative Quality of Recovery Scale. Before surgery (D0), patients with poor quality of recovery had median Visual Analog Scale scores in EuroQol similar to those of patients without poor quality of recovery, but they had more problems in the mobility, usual activities, pain/discomfort, and anxiety/depression dimensions. At 3 months after surgery, patients with poor quality of recovery had median Visual Analog Scale scores in EuroQol similar to those of patients without poor quality of recovery, but they maintained more problems in the pain/discomfort dimension. Patients with poor quality of recovery scored significantly higher on the World Health Organization Disability Assessment Schedule 2.0 scale at baseline, although the results were similar at 3 months. Patients with poor quality of recovery had the worst health status at D0. Evaluation at 3 months indicated similar rates of problems in EuroQol (except for pain/discomfort) and World Health Organization Disability Assessment Schedule 2.0 scores were similar.


Sarcocystis spp. infections in naturally infected cats and dogs: levels of sporocyst production and the influence of host, environmental and seasonal factors on the prevalence of infection.
Data obtained from faecal samples from 508 domestic cats and 481 dogs, previously found to have a high frequency of Sarcocystis spp. infections, were further examined to provide information on levels of sporocyst shedding and to ascertain the influence of host, environmental and seasonal factors on the prevalence of naturally acquired infections. Statistical analysis of prevalence data revealed that, of the factors examined, only host age in cats exerted any apparent influence. Infections were significantly more common in cats up to, rather than over, 6 months of age. Although they tended to be greater in dogs than cats, faecal sporocyst counts in both hosts were generally low, with the majority of infected samples containing 200 sporocysts per gram or less. In infected dogs the highest count recorded was 144,600/g with a mean of 2270/g, and in infected cats it was 11,100/g with a mean of 597/g.


Endothelial cell integrins and COX-2: mediators and therapeutic targets of tumor angiogenesis.
Vascular integrins are essential regulators and mediators of physiological and pathological angiogenesis, including tumor angiogenesis. Integrins provide the physical interaction with the extracellular matrix (ECM) necessary for cell adhesion, migration and positioning, and induce signaling events essential for cell survival, proliferation and differentiation. Integrins preferentially expressed on neovascular endothelial cells, such as alphaVbeta3 and alpha5beta1, are considered as relevant targets for anti-angiogenic therapies. Anti-integrin antibodies and small molecular integrin inhibitors suppress angiogenesis and tumor progression in many animal models, and are currently tested in clinical trials as anti-angiogenic agents. Cyclooxygense-2 (COX-2), a key enzyme in the synthesis of prostaglandins and thromboxans, is highly up-regulated in tumor cells, stromal cells and angiogenic endothelial cells during tumor progression. Recent experiments have demonstrated that COX-2 promotes tumor angiogenesis. Chronic intake of nonsteroidal anti-inflammatory drugs and COX-2 inhibitors significantly reduces the risk of cancer development, and this effect may be due, at least in part, to the inhibition of tumor angiogenesis. Endothelial cell COX-2 promotes integrin alphaVbeta3-mediated endothelial cell adhesion, spreading, migration and angiogenesis through the prostaglandin-cAMP-PKA-dependent activation of the small GTPase Rac. In this article, we review the role of integrins and COX-2 in angiogenesis, their cross talk, and discuss implications relevant to their targeting to suppress tumor angiogenesis.


Biochemical and cellular properties of insulin receptor signalling.
The mechanism of insulin action is a central theme in biology and medicine. In addition to the rather rare condition of insulin deficiency caused by autoimmune destruction of pancreatic β-cells, genetic and acquired abnormalities of insulin action underlie the far more common conditions of type 2 diabetes, obesity and insulin resistance. The latter predisposes to diseases ranging from hypertension to Alzheimer disease and cancer. Hence, understanding the biochemical and cellular properties of insulin receptor signalling is arguably a priority in biomedical research. In the past decade, major progress has led to the delineation of mechanisms of glucose transport, lipid synthesis, storage and mobilization. In addition to direct effects of insulin on signalling kinases and metabolic enzymes, the discovery of mechanisms of insulin-regulated gene transcription has led to a reassessment of the general principles of insulin action. These advances will accelerate the discovery of new treatment modalities for diabetes.


A Systematic Study into the Factors that Affect the Predictive Accuracy of Multilevel VAR(1) Models.
The use of multilevel VAR(1) models to unravel within-individual process dynamics is gaining momentum in psychological research. These models accommodate the structure of intensive longitudinal datasets in which repeated measurements are nested within individuals. They estimate within-individual auto- and cross-regressive relationships while incorporating and using information about the distributions of these effects across individuals. An important quality feature of the obtained estimates pertains to how well they generalize to unseen data. Bulteel and colleagues (Psychol Methods 23(4):740-756, 2018a) showed that this feature can be assessed through a cross-validation approach, yielding a predictive accuracy measure. In this article, we follow up on their results, by performing three simulation studies that allow to systematically study five factors that likely affect the predictive accuracy of multilevel VAR(1) models: (i) the number of measurement occasions per person, (ii) the number of persons, (iii) the number of variables, (iv) the contemporaneous collinearity between the variables, and (v) the distributional shape of the individual differences in the VAR(1) parameters (i.e., normal versus multimodal distributions). Simulation results show that pooling information across individuals and using multilevel techniques prevent overfitting. Also, we show that when variables are expected to show strong contemporaneous correlations, performing multilevel VAR(1) in a reduced variable space can be useful. Furthermore, results reveal that multilevel VAR(1) models with random effects have a better predictive performance than person-specific VAR(1) models when the sample includes groups of individuals that share similar dynamics.


Sleep-disordered breathing in neurodegenerative diseases.
Sleep disorders are common in neurodegenerative diseases such as Parkinson's disease (PD), multiple system atrophy (MSA), amyotrophic lateral sclerosis (ALS), hereditary ataxias, and Alzheimer's disease (AD). Type, frequency, and severity of sleep disturbances vary depending on each of these diseases. Cell loss of the brainstem nuclei that modulates respiration, and dysfunction of bulbar and diaphragmatic muscles increase the risk for sleep-disordered breathing (SDB) in MSA and ALS. The most relevant SDB in MSA is stridor, whereas in ALS nocturnal hypoventilation due to diaphragmatic weakness is the most common sleep breathing abnormality. Stridor and nocturnal hypoventilation are associated with reduced survival in MSA and ALS. In contrast, sleep apnea seems not to be more prevalent in PD than in the general population. In some PD patients, however, coincidental obstructive sleep apnea (OSA) can be the cause of excessive daytime sleepiness (EDS). SDB can also occur in some hereditary ataxias, such as stridor in spinocerebellar ataxia type 3 (Machado-Joseph disease). The presence of concomitant OSA in patients with AD can have deleterious effects on nocturnal sleep, may result in EDS, and might aggravate the cognitive deficits inherent to the disease. However, whether OSA is more frequent in patients with AD than in the general population is uncertain. Recognition of SDB in neurodegenerative disease is important because they are associated with significant morbidity and potential effective treatments are available.


Functional analysis of cyclic diguanylate-modulating proteins in Vibrio fischeri.
As bacterial symbionts transition from a motile free-living state to a sessile biofilm state, they must coordinate behavior changes suitable to each lifestyle. Cyclic diguanylate (c-di-GMP) is an intracellular signaling molecule that can regulate this transition, and it is synthesized by diguanylate cyclase (DGC) enzymes and degraded by phosphodiesterase (PDE) enzymes. Generally, c-di-GMP inhibits motility and promotes biofilm formation. While c-di-GMP and the enzymes that contribute to its metabolism have been well-studied in pathogens, considerably less focus has been placed on c-di-GMP regulation in beneficial symbionts. Vibrio fischeri is the sole beneficial symbiont of the Hawaiian bobtail squid (Euprymna scolopes) light organ, and the bacterium requires both motility and biofilm formation to efficiently colonize. C-di-GMP regulates swimming motility and cellulose exopolysaccharide production in V. fischeri. The genome encodes 50 DGCs and PDEs, and while a few of these proteins have been characterized, the majority have not undergone comprehensive characterization. In this study, we use protein overexpression to systematically characterize the functional potential of all 50 V. fischeri proteins. All 28 predicted DGCs and 14 predicted PDEs displayed at least one phenotype consistent with their predicted function, and a majority of each displayed multiple phenotypes. Finally, active site mutant analysis of proteins with the potential for both DGC and PDE activities revealed potential activities for these proteins. This work presents a systems-level functional analysis of a family of signaling proteins in a tractable animal symbiont and will inform future efforts to characterize the roles of individual proteins during lifestyle transitions.


F-18 Fluorodeoxyglucose positron emission tomography/computed tomography in tuberculosis of the hip: A case report and brief review of literature.
Tuberculosis (TB) has become a global health concern. Cross-sectional imaging modalities like ultrasound, computed tomography and magnetic resonance imaging play an important role in the diagnosis, assessment of disease extent and response to treatment. PET/CT, a unique molecular imaging technique, allows the most accurate correlation of anatomic and metabolic information. We report a case wherein PET/CT played a significant role and contributed valuable information in the evaluation of a patient with TB of the hip. A brief review of the existing literature on the role of PET/CT in musculoskeletal TB is also discussed.


In vitro differences between the lymphocytes of normal subjects and atopics.
Recently evidence has accumulated that atopic disease is associated with a deficiency of thymus-derived (T) cells. This deficiency appears to be primary, rather than secondary to treatment or manifestations of the disease. Results of in vitro studies indicate that the deficiency is most pronounced in certain subpopulations of T cells, and therefore a disturbance of the balance between subsets of T cells, notably suppressor and helper T cells, may develop in vivo in atopics. Some results suggest that there is indeed a relative deficiency of suppressor T cells in atopic diseases, and thus the hyperproduction of IgE which is associated with these diseases may be explained. The cause of the T cell deficiency in atopy may be a basal cellular abnormality, manifested e.g. as increased sensitivity to inactivation by physiological substances, notably to agents which increase intracellular levels of cyclic AMP. It is equally possible that the T cell deficiency is a direct consequence of subnormal production of certain thymic hormones.


Intralesional injection of Pingyangmycin plus corticosteroids may be an effective treatment for cheilitis granulomatosa.
Cheilitis granulomatosa is a rare disease characterised by the recurrent labial swelling of one or both lips with the possibility of the condition to remain on a permanent basis. This kind of granulomatous, inflammatory disease may appear independently or be linked to a paralysis such as the facial and lingua plicata which then characteristic of the Melkersson-Rosenthal syndrome. Classically, a non-necrotizing granulomatous inflammation is seen at histologic examination. Many treatments have been tried. But as its high recurrence rate, the results were often disappointing. The major active component of Pingyangmycin is bleomycin A5. Currently, bleomycin has been proved to be a widely accepted treatment protocol for hemangiomas and lymphangioma. It has been proved that bleomycin could infiltrate prominently into lymphocytes and other inflammatory cells. Stromal connective tissue would proliferate markedly as the result of sclerotherapy. Oedema, lymphangiectasia and perivascular lymphocytic infiltration are always seen in cheilitis granulomatosa histologically. Therefore, we hypothesize that intralesional injection of Pingyangmycin plus corticosteroids may be a safe and effective treatment for cheilitis granulomatosa. The efficacy of this treatment modality is worthy of further investigation.


Hemopoietins for eosinophils. Glycoprotein hormones that regulate the development of inflammation in eosinophilia-associated disease.
Granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin-3 (IL-3), and interleukin-5 (IL-5) are T lymphocyte-derived glycoproteins that stimulate the development of eosinophils from bone marrow precursors. These eosinophil hemopoietins also regulate functions of mature eosinophils, enhancing their ability to infiltrate tissues and to secrete biologically active substances at the tissue site. In these ways, the eosinophil hemopoietins have a major impact on the development of eosinophilia-associated disease.


Funneling before and after anti-incontinence surgery--a prognostic indicator? Part 2: tension-free vaginal tape.
Does the presence of pre- and postoperative funneling of the proximal urethra affect the outcome of tension-free vaginal tape (TVT)? This prospective study of 191 women with proven primary genuine stress urinary incontinence treated by TVT insertion and assessed by pre- and postoperative introital ultrasound aims to answer that question. At 36-month follow-up, the cure rate was 89.5% (Kaplan-Meier estimator). At 6-month follow-up, 6.3% of the women had urge symptoms and 0.5% de novo urge incontinence. The cure rate in preoperative funneling was 77.5% as opposed to 96.6% in the group without funneling (P<0.0001). The funneling rate significantly decreased from 37.2% preoperatively to 17.3% immediately postoperatively (P<0.0001). The continence rate was 57.5% in persistent postoperative funneling vs 96.2% in the group without postoperative funneling (P<0.0001). The complication rate was not higher in the group with persistent postoperative funneling. The TVT procedure significantly reduces bladder neck funneling. Both preoperative and persistent postoperative funneling are associated with an increased probability of therapeutic failure or recurrence.


Amino acid variability, tradeoffs and optimality in human diet.
Studies at the molecular level demonstrate that dietary amino acid intake produces substantial effects on health and disease by modulating metabolism. However, how these effects may manifest in human food consumption and dietary patterns is unknown. Here, we develop a series of algorithms to map, characterize and model the landscape of amino acid content in human food, dietary patterns, and individual consumption including relations to health status, covering over 2,000 foods, ten dietary patterns, and over 30,000 dietary profiles. We find that the type of amino acids contained in foods and human consumption is highly dynamic with variability far exceeding that of fat and carbohydrate. Some amino acids positively associate with conditions such as obesity while others contained in the same food negatively link to disease. Using linear programming and machine learning, we show that these health trade-offs can be accounted for to satisfy biochemical constraints in food and human eating patterns to construct a Pareto front in dietary practice, a means of achieving optimality in the face of trade-offs that are commonly considered in economic and evolutionary theories. Thus this study may enable the design of human protein quality intake guidelines based on a quantitative framework.


GC-MS and GC-IRD analysis of 2-, 3- and 4-methylmethamphetamine and 2-, 3- and 4-methylamphetamine.
4-Methylmethamphetamine has been detected in samples submitted for analysis in several states throughout Australia. Six ring substituted methyl isomers of methamphetamine and amphetamine were synthesised and analysed. As the regioisomeric 2-, 3- and 4-methylmethamphetamine and 2-, 3- and 4-methylamphetamine have virtually identical mass spectra, the use of MS is an ineffective technique to discriminate between these closely related compounds. We set out to determine whether the regioisomers could be differentiated by a combination of GC-MS, acetyl derivatisation and GC-IRD. We demonstrate that the three isomers of methylmethamphetamine and methylamphetamine can be separated by GC, and a combination of acetyl derivatisation and vapour phase IR can identify the specific ring substituted compound.


Microsurgical anastomosis of vas deferens by a telescopic technique.
A telescopic microsurgical anastomosis of the vas deferens was performed on 24 rats. Sequential histological examination demonstrated mucosal healing in 7 days. Healing was complete in 21 days, and tubal patency was confirmed by histology in 100% of the cases.


Hydatid cysts: does every picture tell a story?
Ultrasound imaging is an effective aid to the detection of abdominal cystic echinococcosis. Recently, the images produced have provided more information than just a diagnosis. Different cyst morphologies have been classified into types, and such classifications can be considered to represent a natural history of cyst development. Understanding how cyst structures alter with time is crucial to evaluating the effectiveness of different treatment regimes and to understanding natural degeneration of cysts that can occur. However, the developmental events seen in ultrasound need to be matched more closely to immunological and cellular events if we are to have a fuller appreciation of disease progression and regression. At a community level, the relative proportions of cyst types could also provide epidemiological information on disease transmission.


Emergence of novel subtypes of Cryptosporidium parvum in calves in Poland.
The aim of the study was to identify the Cryptosporidium parvum subtypes circulating in Polish cattle and their distribution in relation to the age and health status of tested animals. In total, 779 fecal samples were obtained from young cattle originating from 237 farms. C. parvum strains were identified at the 18 small-subunit ribosomal RNA (SSU rRNA), COWP, and LIB13 loci and were subsequently analyzed by sequencing at the 60-kDa glycoprotein (GP60) locus for subtype determination. The presence of 71 C. parvum strains belonging to IIa, IId, or IIl subtype families was shown. The strains from the IIa allele family prevailed with IIaA17G1R1, IIaA17G2R1, and IIaA15G2R1 subtypes occurring frequently. Two novel subtypes IIaA10G1R1 and IIlA19R3 were detected for the first time in a bovine host. The highest C. parvum prevalence (22.5 %, confidence interval (CI) = 2.5 %) was observed among the youngest animals up to 2 weeks of age, followed by the prevalence among those aged 2 to 4 weeks (6.6 %, CI = 2.6 %) and then among older cattle (4.9 %, CI = 2.1). The occurrence of diarrhea in animals was associated with the presence of the IIaA16G1R1b subtype, while infections caused by IIaA15G2R1 strains were more likely to be asymptomatic. The geographical distribution of subtypes revealed that strains from the IIa subtype family were detected all over the country frequently compared to the IId and IIl subtypes, the sporadic appearances of which confirmed their endemic occurrence. Subtype analysis revealed the presence of zoonotic strains indicating cattle as a reservoir for human cryptosporidiosis.


[PROGRESS OF MESENCHYMAL STEM CELL-DERIVED Exosomes IN TISSUE REPAIR].
To comprehensively analyze the recent advancements in the field of mesenchymal stem cells (MSCs) derived exosomes (MSCs-exosomes) in tissue repair. The literature about MSCs-exosomes in tissue repair was reviewed and analyzed. Exosomes are biologically active microvesicles released from MSCs which are loaded with functional proteins, RNA, and microRNA. Exosomes can inhibit apoptosis, stimulate proliferation, alter cell phenotype in tissue repair of several diseases through cell-to-cell communication. MSCs-exosomes is a novel source for the treatment of tissue repair. Further research of MSCs-exosomes biofunction, paracellular transport, and treatment mechanism will help the transform to clinical application.


Assessment of left ventricular hypertrophy in a trained athlete: differential diagnosis of physiologic athlete's heart from pathologic hypertrophy.
Physiologic LV remodeling in young trained athletes as a consequence of chronic training can occasionally mimic certain pathologic conditions associated with sudden death, such as HCM. A small but important subset of elite male athletes may show a borderline increased LV wall thickness of 13 to 15 mm, which defines a gray zone of overlap between the extreme expressions of athlete's heart and a mild HCM phenotype. Such diagnostic ambiguity can be resolved by using the paradigm of noninvasive parameters including testing with echocardiography (and, more recently, with CMR): left atrial and LV chamber dimensions and shape, brief periods of deconditioning to alter LV mass, measurement of oxygen consumption and diastolic filling, and recognition of familial occurrence of HCM or a pathogenic HCM-causing sarcomere mutation. Such distinctions between physiologic/benign athlete's heart and HCM, the most common cause of sudden death in the young in the United States, can be crucial. The recognition of HCM leads to disqualification from intense competitive sports to reduce sudden death risk and, when appropriate, permits initiation of therapeutic interventions.


Accelerating eye movement research via accurate and affordable smartphone eye tracking.
Eye tracking has been widely used for decades in vision research, language and usability. However, most prior research has focused on large desktop displays using specialized eye trackers that are expensive and cannot scale. Little is known about eye movement behavior on phones, despite their pervasiveness and large amount of time spent. We leverage machine learning to demonstrate accurate smartphone-based eye tracking without any additional hardware. We show that the accuracy of our method is comparable to state-of-the-art mobile eye trackers that are 100x more expensive. Using data from over 100 opted-in users, we replicate key findings from previous eye movement research on oculomotor tasks and saliency analyses during natural image viewing. In addition, we demonstrate the utility of smartphone-based gaze for detecting reading comprehension difficulty. Our results show the potential for scaling eye movement research by orders-of-magnitude to thousands of participants (with explicit consent), enabling advances in vision research, accessibility and healthcare.


Impact of the COVID-19 pandemic on the mental and physical health and overall wellbeing of university students in Portugal.
Throughout the pandemic of COVID-19 caused by SARS-CoV-2, university students were considered a vulnerable risk group for mental health impairment and wellbeing deterioration. This study aimed at evaluating the pandemic's impact on the physical and mental health and wellbeing among students of a Portuguese university. This cross-sectional study included 913 participants and ran from June to October 2020. Data collected included sociodemographics, three mental health self-report questionnaires (Depression Anxiety Stress Scale, Eating Disorder Examination Questionnaire and Brief COPE) and lifestyle practices (eating and sleeping patterns, media, and entertainment habits) during the first months of the pandemic, which included a 72-day full national lockdown. Descriptive and correlational statistical analysis were conducted. Students' food habits changed during the pandemic, namely on the consumption of snacks and fast food and, overall, less balanced meals became more prevalent. Additionally, almost 70% of the students reported Body Mass Index changes, while 59% went through sleep pattern changes-these were more pronounced in women and younger students. Over half (67%) of the inquirees exhibited an increase in their stress, depression, and generalized anxiety symptoms. Also, the study demonstrates that students' lifestyles trended negatively during the pandemic and highlights how important regular psychological, health monitoring and emotional support is, amongst this somehow overlooked population throughout the pandemic. Universities should provide support to overcome challenges in future stressful situations. This study might have an impact on how universities and higher education systems approach their students in terms of mental and physical health monitoring and promotion in future situations, non-related with COVID. Moreover, it has a large sample of students well characterized in terms of mental and physical health, which might be of interest for future comparison with other worldwide group of students throughout stressful situations, such as tragic events, wars, pandemics.


Electroencephalographic sleep profiles in single-episode and recurrent unipolar forms of major depression: II. Comparison during remission.
Previous studies indicate that recurrent forms of depression are associated with greater biological disturbances as compared to single-episode cases. This study examines whether the observed differences in the sleep patterns during recurrent and single-episode depression persist into remission following nonpharmacologic treatment. Two groups of patients (27 single episode [SE] and 53 recurrent unipolar [RU]) with major depression underwent sleep studies before and after nonpharmacologic treatment. Groups were equated for age, severity, and proportion of men and women. Groups were compared using multivariate analyses of covariance and/or analyses of covariance to examine six sets of sleep measures. The differences observed between the SE and RU groups during the index episode persisted into early remission. The findings of greater disturbances of sleep continuity, rapid eye movement sleep and diminished slow wave sleep in the RU group supports the hypothesis that recurrent depression is associated with a more severe neurophysiological substrate than clinically comparable SE cases. Although these observations are consistent with an illness progression model, the possibility that recurrent affective illness is associated with a more virulent, stable phenotype cannot be ruled out. Resolution of this issue requires longitudinal and family studies.


An ionic liquid-in-water microemulsion as a potential carrier for topical delivery of poorly water soluble drug: Development, ex-vivo and in-vivo evaluation.
In this paper, we report an ionic liquid-in-water (IL/w) microemulsion (ME) formulation which is able to solubilize etodolac (ETO), a poorly water soluble drug for topical delivery using BMIMPF6 (1-butyl-3-methylimidazolium hexafluorophosphate) as IL, Tween 80 as surfactant and ethanol as co-surfactant. The prepared ME was characterized for physicochemical parameters, subjected to ex-vivo permeation studies as well as in-vivo pharmacodynamic evaluation. The ex-vivo drug permeation studies through rat skin was performed using Franz-diffusion cell and the IL/w based ME showed maximum mean cumulative percent permeation of 99.030±0.921% in comparison to oil-in-water (o/w) ME (61.548±1.875%) and oily solution (48.830±2.488%) of ETO. In-vivo anti-arthritic and anti-inflammatory activities of the prepared formulations were evaluated using different rodent models and the results revealed that ETO loaded IL/w based ME was found to be more effective in controlling inflammation than oily solution, o/w ME and marketed formulation of ETO. Histopathological studies also demonstrated that IL/w based ME caused no anatomical and pathological changes in the skin.


Noise figure of electromagnetic systems with parity and time-reversal symmetry.
The primary bottlenecks in designing and implementing PT-symmetric systems at microwave frequency ranges are noise and instability which can weakly break PT-symmetry resulting in system performance degradation. Practical implementation of such systems and devices require significant level of control and stability and it is crucial to analyze the noise performance of such systems in terms of noise figure and signal-to-noise ratio. We describe and develop a simulation model to calculate noise figure of PT-symmetric system and evaluate the performance degradation. We also discuss application design and circuit configurations that could reduce the noise figure resulting in better performance.


Erythroid stem cells in Rauscher virusinfected mice.
Using the technique for erythroid colony formation in vitro, bone marrow and spleen cells from NMRI mice were studied after Rauscher virus (RLV) infection. There was a substantial decrease in CFUE concentrations during the first days after infection, this being more pronounced in the marrow than in the spleen. In the marrow values gradually return to and are maintained at control levels, while in the spleen a 40-50-fold increase is seen between days 8 and 18. In normal and RLV infected animals the same dose dependence of CFUE growth for erythropoietin was seen. For normal and RLV infected cells in the absence of erythropoietin there were only very few background colonies. In exhypoxic plethoric mice the increase in CFUE concentration seen in normal mice in the spleen, is delayed by 2-3 days.


Activation of serotonin receptors in the medial basal hypothalamus stimulates growth hormone secretion in the unanesthetized rat.
In conscious rats, serotonin microinjected into the basal hypothalamus caused secretion of GH maximal within 10-25 min. The effects of serotonin on GH were blocked by the non-selective serotonin receptor blocker, metergoline 2.5 mg/kg, but not by the serotonin type 2 receptor blocker, ketanserine 0.2 mg/kg. Injections of serotonin in the preoptic/anterior hypothalamic area were without effect. It is concluded that activation of serotonin receptors, probably type I, on or near GH releasing factor neurons in the arcuate nucleus causes secretion of GH and that serotonin has no direct effect on or near somatostatin neurons in the preoptic anterior hypothalamic area.


Second primary squamous cell carcinoma arising in a skin flap: a case report and literature review on etiologic factors and treatment strategy.
The number of reports describing malignant tumors arising in flaps has been increasing recently. This report describes the case of a patient who had a second squamous cell carcinoma (SCC) in the center of the skin island of a forearm flap raised approximately 6 years previously. Histopathologic examination suggested human papillomavirus infection and "mucosalization" of the flap. In addition, the authors review previous cases, with special attention to etiologic factors and surgical strategies. There might be a causal relation between second primary SCC in the flap and the carcinogenic effect (chronic inflammation) induced by the environment surrounding the squamous epithelium of the flap. If environmental factors have greatly contributed to the occurrence of a tumor, there is a potential for tumor occurrence in the remaining flap. There also might be a potential for generating second primary SCC in these foci. The results suggested that total resection of the flap might be recommended in the surgical resection of second primary SCC arising in a flap.


Binding and Internalization in Receptor-Targeted Carriers: The Complex Role of CD44 in the Uptake of Hyaluronic Acid-Based Nanoparticles (siRNA Delivery).
CD44 is an endocytic hyaluronic acid (HA) receptor, and is overexpressed in many carcinomas. This has encouraged the use of HA to design CD44-targeting carriers. This paper is about dissecting the mechanistic role of CD44. Here, HA-decorated nanoparticles are used to deliver siRNA to both tumoral (AsPC-1, PANC-1, HT-29, HCT-116) and non-tumoral (fibroblasts, differently polarized THP-1 macrophages, HUVEC) human cell lines, evaluating the initial binding of the nanoparticles, their internalization rate, and the silencing efficiency (cyclophilin B (PPIB) gene). Tumoral cells internalize faster and experience higher silencing than non-tumoral cells. This is promising as it suggests that, in a tumor, HA nanocarriers may have limited off-target effects. More far-reaching is the inter-relation between the four parameters of the study: CD44 expression, HA binding on cell surfaces, internalization rate, and silencing efficiency. No correlation is found between binding (an early event) and any of the other parameters, whereas silencing correlates both with speed of the internalization process and CD44 expression. This study confirms on one hand that HA-based carriers can perform a targeted action, but on the other it suggests that this may not be due to a selective binding event, but rather to a later recognition leading to selective internalization.


Neural correlates of perceptual learning in a sensory-motor, but not a sensory, cortical area.
This study aimed to identify neural mechanisms that underlie perceptual learning in a visual-discrimination task. We trained two monkeys (Macaca mulatta) to determine the direction of visual motion while we recorded from their middle temporal area (MT), which in trained monkeys represents motion information that is used to solve the task, and lateral intraparietal area (LIP), which represents the transformation of motion information into a saccadic choice. During training, improved behavioral sensitivity to weak motion signals was accompanied by changes in motion-driven responses of neurons in LIP, but not in MT. The time course and magnitude of the changes in LIP correlated with the changes in behavioral sensitivity throughout training. Thus, for this task, perceptual learning does not appear to involve improvements in how sensory information is represented in the brain, but rather how the sensory representation is interpreted to form the decision that guides behavior.


Prosthetic contingencies for future tooth loss.
Possible contingencies for future natural tooth loss that can and should be built into prostheses have been reviewed. These contingencies provide a measure of security to the patient and the dentist when attempting to maintain teeth that appear to have a guarded prognosis. Without a contingency these questionable teeth would be extracted early in therapy from fear that a completely new prosthesis would have to be made if they were lost at a later time. In maintaining more natural teeth, the patient has better function, more occlusal and vertical support, longer maintenance of the alveolar ridge, more support for the prosthesis, and more remaining teeth to share all the forces. This approach of building in contingencies for future tooth loss allows the patient and the dentist the freedom to attempt therapy on abutments with a guarded prognosis with little financial risk.


[Pelvic mass in 70 years old patient: Difficult diagnosis of Erdheim-Chester disease].
Erdheim-Chester disease (EC) is a rare disease that is included in Group L in the 2016 revised classification of Langheransian histiocytoses. This disease may be clinically asymptomatic or manifest as a multi-systemic and life-threatening condition. All organs can be affected but typically there is bone involvement, retroperitoneal fibrosis, pituitary involvement, involvement of large vessels, lung, pleura or central nervous system. We are reporting a 70-year-old patient who, as of 2014, had a pelvic mass with retroperitoneal fibrosis and large vessel vasculitis without a definite diagnosis. Histological and molecular examination of the surgical specimen of the pelvic mass with the discovery of the BRAF V600E mutation provided new elements for the definitive diagnosis of Erdheim-Chester disease. We will describe the clinical, histological and molecular features to be known in EC disease.


The genus Artemia, the nanoplastics, the microplastics, and their toxic effects: a review.
Plastic pollution is a threat to the marine environment, the destination of mismanaged plastic. Due to reduced size, microplastics and nanoplastics (MNPs) can interact with a wide range of organisms. Non-selective filter feeder zooplanktonic microcrustaceans are potential targets for MNP accumulation. Zooplankton is a key group for the food web, linking primary producers to secondary consumers. The genus Artemia has been widely used to investigate the effects of plastic particles on the biota. The present work critically reviewed the ecotoxicological studies about plastic particles and Artemia, pointing out methodological aspects and effects caused by MNPs, highlighting their importance and limitations, and suggesting directions for future research. We analyzed twenty-one parameters into four categories: characteristics of plastic particles, general particularities of brine shrimp, methodologies of the cultures, and toxicological parameters. The principal gaps in the area are the lack of methodological standardization regarding the physicochemical parameters of the particles, the biology of the animals, and culture conditions. Even though few studies performed realistic exposure scenarios, results indicate MNPs as potential harmful contaminants to microcrustaceans. The main effects reported were particle ingestion and accumulation followed by reduced brine shrimp survival/mobility. The present review poses Artemia as suitable animals for investigations concerning the risks of MNP exposure at the individual level and to the ecosystems, although protocol standardization is still needed.


A possible role for bcl-2 in regulating T-cell memory--a 'balancing act' between cell death and survival.
One of the central questions in T-cell immunity is how the generation of T-cell memory may proceed simultaneously with the maintenance of T-cell homeostasis. In this article, we present a hypothesis that the regulation of bcl-2 gene expression within the activated CD45RO+ T-cell population may be one of the key factors in the balance between death and survival of these activated mature T cells, both in vitro and in vivo.


4-Chloro-1-methyl-indoline-2,3-dione.
The title mol-ecule, C(9)H(6)ClNO(2), is essentially planar; the maximum deviation of the indoline ring system is 0.027 (3) Å and the substituents do not deviate by more than 0.075 (2) Å from this plane. Inter-molecular C-H⋯O hydrogen bonds consolidate the crystal structure.


Development and use of an infant resuscitation performance tool (Infa-RePT) to improve team performance.
Measurement of neonatal team resuscitation performance is critical to identify opportunities for improvement and to target education. An effective tool to measure team performance during infant resuscitations is lacking. We developed an in-hospital infant resuscitation performance tool (Infa-RePT) using the modified Delphi method. We employed a QI framework and targeted interventions, including the use of role responsibility checklists, mock codes, and an educational video. We tracked Infa-RePT scores, mock code team attendance, and confidence surveys. Our specific aim was to improve Infa-RePT score from a baseline of 7.4 to <5 (lower is better) over a 26-month period. Twenty-five elements reached >80% consensus as essential components to include on the Infa-RePT. Independent observation showed 86% concordance on checklist items. Simulation (n = 26) and unit-based code (n = 10) Infa-RePT scores showed significant improvement after project start from 7.4 to 4.2 (p < 0.01) with special cause variation noted on control chart analysis. No significant difference was observed between simulations and in-unit codes. Staff confidence self-reports improved over the study period. Use of a novel scoring tool can help monitor team progress over time and identify areas for improvement. Focused interventions can improve resuscitation team performance. We developed and used a novel, comprehensive measurement tool for team infant resuscitation performance in both simulation and in-unit settings. Using QI methodology, team performance improved after the enhancement of a mock code simulation program. Review of team performance scores can highlight key areas to target interventions and monitor progress over time.


Dynamic G- and R-banding of human chromosomes for electron microscopy.
Synchronized human lymphocytes were exposed to 5-bromo-2'-deoxyuridine (BrdUrd) for incorporation in either G- or R-bands. The substituted bands were revealed by monoclonal anti-BrdUrd antibodies disclosed with either gold-labeled antibodies or with the protein A-gold complex. Sharp G- or R-banding, specific for electron microscopy (EM), was obtained. These banding patterns, referred to as GB-AAu (G-bands by BrdUrd using Antibodies and gold [Au]) and RB-AAu (R-bands by BrdUrd using Antibodies and gold [Au]), resemble dynamic band patterns (GBG and RBG) much more than they do morphologic band patterns (GTG and RHG). The G- and R-band patterns allow accurate chromosome identification and karyotyping. An actual karyotype of human GB-AAu-banded chromosomes at the 750 band level, photographed in the EM, is presented. The method produces excellent band separation and band contrast. Variations in band staining intensities were noted and correlated with BrdUrd enrichment. The C-band regions were positively stained after GB-AAu banding while they were negatively stained after RB-AAu banding. Telomeres appeared heterogeneous after GB-AAu banding suggesting that part of the telomeric bands might be late replicating.


Tracheostomaplasty: A surgical method for improving retention of an intraluminal stoma button for hands-free tracheoesophageal speech.
We describe a minimally invasive surgical technique, tracheostomaplasty, to overcome anatomical deformities of the stoma that preclude successful retention of a stoma button for hands free tracheoesophageal (TE) speech. We conducted a retrospective analysis of 21 patients who underwent tracheostomaplasty after laryngectomy to accommodate an intraluminal valve attachment for hands-free TE speech. Sixteen men and 5 women (median age, 65 years; median follow-up, 27.7 months) underwent tracheostomaplasty; 6 patients developed a mild cellulitis that required therapy and 5 patients required a minor revision surgery. At last follow-up, 15 (71%) patients successfully achieved hands-free TE speech using an intraluminal stoma button. Three patients only retained the intraluminal device to facilitate digital occlusion. Tracheostomaplasty failed in 3 patients because of granulation tissue formation or stomal stenosis. Tracheostomaplasty is a successful technique to improve intraluminal retention of a stoma button for hands-free TE speech in laryngectomy patients.


[Dependence of lipid peroxidation on the typologic characteristics of the nervous system and their relation to the resistance of the body to exertion].
Resistance of animals with different type of the nervous activity to physical exercises depends on the activity of lipid peroxidation processes in tissues. The highest degree of adaptation to prolonged running is observed in the inhibitory-type rats as against the excitable and even-tempered ones.


Identification and visualization of oxidized lipids in atherosclerotic plaques by microscopic imaging mass spectrometry-based metabolomics.
Dysregulated lipid metabolism has emerged as one of the major risk factors of atherosclerosis. Presently, there is a consensus that oxidized LDL (oxLDL) promotes development of atherosclerosis and downstream chronic inflammatory responses. Due to the dynamic metabolic disposition of lipoprotein, conventional approach to purify bioactive lipids for subsequent comprehensive analysis has proven to be inadequate for elucidation of the oxidized lipids species accountable for pathophysiology of atherosclerotic lesions. Herein, we aimed to utilize a novel mass microscopic imaging technology, coupled with mass spectrometry (MS) to characterize oxidized lipids in atherosclerotic lesions. We attempted to use MALDI-TOF-MS and iMScope to identify selected oxidized lipid targets and visualize their respective localizations in study models of atherosclerosis. Based on the MS analysis, detection of 7-K under positive ionization through product ion peak at m/z 383 [M + H-H2O] indicated the distinctive presence of targeted lipid within Cu2+-oxLDL and Cu2+-oxLDL loaded macrophage-like J774A.1 cells, along with other cholesterol oxidation products. Moreover, the application of two-dimensional iMScope has successfully visualized the localization of lipids in aortic atherosclerotic plaques of the Watanabe heritable hyperlipidemic (WHHL) rabbit. Distinctive lipid distribution profiles were observed in atherosclerotic lesions of different sizes, especially the localizations of lysoPCs in atherosclerotic plaques. Taken together, we believe that both MALDI-TOF-MS and iMScope metabolomics technology may offer a novel proposition for future pathophysiological studies of lipid metabolism in atherosclerosis.


The unstable shoulder: recurring subluxation.
A review of operative repairs for recurring dislocation of the shoulder revealed that a significant proportion of these operations was being done for what could best be described as recurring subluxation of the shoulder. The presenting complaint was of the shoulder 'going out of joint', but no significant trauma was recalled, dislocation was never shown on the radiograph and none required manual reduction. The only physical finding was apprehension on external rotation of the shoulder in abduction. Radiographs were frequently normal and arthrography and cineradiography were not helpful in confirming anterior displacement. The most useful preoperative information was obtained by manipulation of the shoulder under general anaesthesia just before the surgical repair. In all instances anterior instability could be demonstrated. Of 99 Magnuson-Stack repairs drawn from the records of the Vancouver General Hospital in a 3-year period, 34 proved to be examples of recurring subluxation. This high proportion of such patients contradicts the teaching in standard orthopaedic textbooks, but substantiates the warning of Rowe (1963) to beware of the patient whose shoulder 'dislocates' initially with little evidence of injury. It also substantiates Saha's concept (1971) of inherent shoulder instability as a contributor to the incidence of recurring dislocation.


Multi-criteria decision analysis for the evaluation and screening of sustainable aviation fuel production pathways.
The aviation sector, a significant greenhouse gas emitter, must lower its emissions to alleviate the climate change impact. Decarbonization can be achieved by converting low-carbon feedstock to sustainable aviation fuel (SAF). This study reviews SAF production pathways like hydroprocessed esters and fatty acids (HEFA), gasification and Fischer-Tropsch Process (GFT), Alcohol to Jet (ATJ), direct sugar to hydrocarbon (DSHC), and fast pyrolysis (FP). Each pathway's advantages, limitations, cost-effectiveness, and environmental impact are detailed, with reaction pathways, feedstock, and catalyst requirements. A multi-criteria decision framework (MCDS) was used to rank the most promising SAF production pathways. The results show the performance ranking order as HEFA > DSHC > FP > ATJ > GFT, assuming equal weight for all criteria.


Atlantooccipital Overlap and Other Craniocervical Junction Abnormalities in Dogs.
The term craniocervical junction abnormality (CJA) is an umbrella term for a variety of malformations that occur in the craniocervical region. These abnormalities include Chiari-like malformation, atlantooccipital overlapping, atlantoaxial instability, occipitoatlantoaxial malformations, atlantoaxial dural bands, and dens abnormalities. Syringomyelia can occur secondary to any of these malformations or a combination. Clinical signs in dogs with a CJA typically include neck pain and varying signs associated with a cervical myelopathy. MRI of CJAs with computed tomography imaging is necessary for a full evaluation. Some dogs with CJAs respond to medical management, but for most surgery is the treatment of choice.


Efficacy of intramuscular and intraperitoneal deferoxamine for aluminum chelation.
As intravenous administration of deferoxamine is difficult in home dialysis patients we set out to determine the efficacy of intramuscular (i.m.) and intraperitoneal (i.p.) deferoxamine for removal of aluminum. Patients with serum aluminum levels greater than 90 micrograms/liter were studied in a paired fashion with each patient serving as their own control. Serum and peritoneal fluid aluminum were determined using flameless atomic absorption. In hemodialysis patients 2 g of intravenous deferoxamine increased serum aluminum from 124.7 +/- 32.4 to 415 +/- 192.4 micrograms/liter. One g of deferoxamine given intravenously or intramuscularly resulted in 76.8 +/- 35.3% and 70.4 +/- 23.2%, respectively, of the 2 g i.v. response. The rate at which serum aluminum increased following i.v. deferoxamine infusion was biphasic, with an initial rapid phase lasting 139 minutes followed by a much slower phase. The volume of distribution of aluminum following deferoxamine administration was 12.6 +/- 1.61 and the half life (t1/2) for aluminum removal during hemodialysis was 9.0 +/- 2.0 hours. The increase in serum aluminum following deferoxamine was not due to chelation of erythrocyte aluminum as erythrocyte aluminum remained constant over 24 hours. In patients on continuous ambulatory peritoneal dialysis, 2 g intravenous deferoxamine resulted in the removal of 560 +/- 267 micrograms of aluminum over 24 hours while 2 g deferoxamine given intraperitoneally gave 91 +/- 13% of the intravenous response. Aluminum clearance over 48 hours was twice that for 24 hours for both i.v. and i.p. deferoxamine.(ABSTRACT TRUNCATED AT 250 WORDS)


Quantitative Modeling of Nucleosome Unwrapping from Both Ends.
In eukaryotic cells, DNA is packaged into chromatin where nucleosomes are the basic packaging unit. Important cellular processes including gene expression, DNA replication, and DNA repair require nucleosomal DNA to be unwrapped so that functional proteins can access their target sites, which otherwise are sterically occluded. A key question in this process is what the unwrapped conformations individual nucleosomes adopt within chromatin are. Here, we develop a concurrent nucleosome unwrapping model to address this question. We hypothesize that for a given end-to-end distance of the nucleosomal DNA, the nucleosomal DNA stochastically unwraps from the histone core from both ends independently and that this combination of unwrapping from both sides results in a significant increase in the average distance between the DNA extending from both sides of the nucleosomes. We test our model on recently published experiments using a DNA origami nanocaliper that quantifies nucleosome unwrapping and achieve good agreement between experiment and model prediction. We then investigate the DNA origami caliper distribution when attached to a hexasome (a nucleosome lacking an H2A/H2B dimer). A significant shift in the caliper angle distribution caused by the asymmetric structural features of the hexasome seen experimentally is consistent with the model. Our modeling approach may be more broadly useful to the interpretation of other studies of nucleosome dynamics, chromatin dynamics, and regulatory processes involving nucleosome unwrapping, as well as more generally to optimization of future DNA origami designs to probe mechanical properties of biomolecules.


Transcriptomic analysis of Malpighian tubules from the stingless bee Melipona scutellaris reveals thiamethoxam-induced damages.
The concern about pesticide exposure to neotropical bees has been increasing in the last few years, and knowledge gaps have been identified. Although stingless bees, (e.g.: Melipona scutellaris), are more diverse than honeybees and they stand out in the pollination of several valuable economical crops, toxicity assessments with stingless bees are still scarce. Nowadays new approaches in ecotoxicological studies, such as omic analysis, were pointed out as a strategy to reveal mechanisms of how bees deal with these stressors. To date, no molecular techniques have been applied for the evaluation of target and/or non-target organs in stingless bees, such as the Malpighian tubules (Mt). Therefore, in the present study, we evaluated the differentially expressed genes (DEGs) in the Mt of M. scutellaris after one and eight days of exposure to LC50/100 (0.000543 ng a.i./μL) of thiamethoxam (TMX). Through functional annotation analysis of four transcriptome libraries, the time course line approach revealed 237 DEGs (nine clusters) associated with carbon/energy metabolism and cellular processes (lysosomes, autophagy, and glycan degradation). The expression profiles of Mt were altered by TMX in processes, such as detoxification, excretion, tissue regeneration, oxidative stress, apoptosis, and DNA repair. Transcriptome analysis showed that cell metabolism in Mt was mainly affected after 8 days of exposure. Nine genes were selected from different clusters and validated by RT-qPCR. According to our findings, TMX promotes several types of damage in Mt cells at the molecular level. Therefore, interference of different cellular processes directly affects the health of M. scutellaris by compromising the function of Mt.


Short-term effects of chewing gum on satiety and afternoon snack intake in healthy weight and obese women.
Afternoon snacking contributes significantly to total energy intake. Strategies to enhance the satiety value of lunch and reduce afternoon snacking are of interest for body weight management. To assess whether between-meal gum chewing would enhance the satiety response to a fixed lunch meal; and assess the role of cholecystokinin (CCK) as a potential mediator of the response in non-obese healthy weight and obese women. Fifty unrestrained obese (n=25) and non-obese healthy weight (n=25) women participated in a two-arm cross-over study assessing multiple (15min per hour×3h) gum chewing (GUM) occurrences or no gum (Control) on subjective ratings of satiety, subsequent sweet and salty snack intake, CCK and general metabolic responses. GUM compared to Control resulted in significant suppression of hunger, desire to eat and prospective consumption (p<0.05). Total snack energy intake was reduced ~9.3% by GUM, but not significantly different from Control (p=0.08). However, overall carbohydrate intake was reduced by GUM (p=0.03). This was consistent with a reduction in snacks characterized as high carbohydrate, low fat (p=0.02). BMI specific effects indicated GUM reduced pretzel intake in obese women (p=0.05) and Oreo cookie intake in healthy weight women (p=0.03) 3h after lunch. Metabolic responses and CCK did not differ between experimental conditions. Chewing gum intermittently post-lunch enhances perceptions of satiety and may have important implications in reducing afternoon high carbohydrate-snack intake.


The expression of transthyretin mRNA in the developing rat brain.
Specific cDNA and oligonucleotide probes were used to study the appearance of transthyretin mRNA in developing rat brain using Northern gel analysis, cytoplasmic dot hybridization, and in situ hybridization. Transthyretin mRNA in embryonic rat brain was found to be confined to the epithelial layer of the choroid plexus primordia appearing first in the fourth ventricle, followed by appearance in the lateral ventricles, and subsequently in the third ventricle. Transthyretin mRNA was localized in these cells from early stages of neuroepithelium differentiation, showing that it is a sensitive marker for the differentiation of the choroid plexus within the fetal brain.


Voice evaluation of myomucosal shunt after total laryngectomy: comparison with esophageal speech.
The vocal quality attained with a tracheoesophageal myomucosal shunt (MMS) as described by Strome was evaluated in four patients and compared with three esophageal speakers and two normal subjects. The patients with MMSs acquired speech sooner. Fundamental frequency, pitch, timbre, and melody were analyzed with computerized electroglottography and sonography. Intelligibility was deemed better after the MMS primarily because phonation time approximated that of normal speech, and this study suggests that, following total laryngectomy, the vocal quality achieved using the MMS is preferrable to that of esophageal speech.


The negative effects of litter of parent plants of Cirsium vulgare on their offspring: autotoxicity or immobilization?
It is shown that litter of flowering plants of Cirsium vulgare inhibits the growth of seedlings of the same species at concentrations as low as 0.4% (g litter/g soil). The inhibition of growth cannot be fully compensated by adding nutrients, which indicates that autotoxicity may occur. It must be concluded, however, from an analysis of the results that immobilization of nutrients by microorganisms is the main cause of growth reduction if litter is added to the soil. In a field experiment under nutrient rich conditions plant litter did not affect germination, survival or growth of seedlings. The hypothesis is put forward that under conditions of nutrient deficiency and at high population densities, immobilization of nitrogen by Cirsium vulgare litter may influence plant growth. Results are discussed in relation to those of Stachon and Zimdahl (1980) and Wilson (1981) who claimed allelopathy for the related Cirsium arvense.


[On the technique of applying drugs in solid form to rats (author's transl)].
A method is described which allows the oral administration of drugs in solid form by hardgelatin mini-caps to rats with a simple device. The method is especially suitable for the administration of solid, radioactively labelled drugs.


Expression of zonula occludens-1 (ZO-1) and the transcription factor ZO-1-associated nucleic acid-binding protein (ZONAB)-MsY3 in glial cells and colocalization at oligodendrocyte and astrocyte gap junctions in mouse brain.
The PDZ domain-containing protein zonula occludens-1 (ZO-1) interacts with several members of the connexin (Cx) family of gap junction-forming proteins and has been localized to gap junctions, including those containing Cx47 in oligodendrocytes. We now provide evidence for ZO-1 expression in astrocytes in vivo and association with astrocytic connexins by confocal immunofluorescence demonstration of ZO-1 colocalization with astrocytic Cx30 and Cx43, and by ZO-1 coimmunoprecipitation with Cx30 and Cx43. Evidence for direct interaction of Cx30 with ZO-1 was obtained by pull-down assays that indicated binding of Cx30 to the second of the three PDZ domains in ZO-1. Further, we investigated mouse Y-box transcription factor MsY3, the canine ortholog of which has been termed ZO-1-associated nucleic acid-binding protein (ZONAB) and previously reported to interact with ZO-1. By immunofluorescence using specific antimouse ZONAB antibody, ZONAB was found to be associated with oligodendrocytes throughout mouse brain and spinal cord, and to be colocalized with oligodendrocytic Cx47 and Cx32 as well as with astrocytic Cx43. Our results extend the CNS cell types that express the multifunctional protein ZO-1, demonstrate an additional connexin (Cx30) that directly interacts with ZO-1, and show for the first time the association of a transcription factor (ZONAB) with ZO-1 localized to oligodendrocyte and astrocyte gap junctions. Given previous observations that ZONAB and ZO-1 in combination regulate gene expression, our results suggest roles of glial gap junction-mediated anchoring of signalling molecules in a wide variety of glial homeostatic processes.


Biological and chemical studies on overheated brewed coffee.
Vapour formed from overheated decaffeinated coffee was condensed and tested for mutagenicity using the Ames assay in Salmonella typhimurium strains TA98 and TA100. Vapour produced at 73 and 100 degrees C exhibited no mutagenicity. The basic fraction of vapour produced at 350 degrees C showed weak mutagenicity towards strains TA98 with metabolic activation. The chemical analysis of this fraction identified pyridines and pyrazines as the major constituents. None of the compounds identified in this fraction has been reported as mutagenic when tested in the Ames assay.


Activation of protein kinase A by atrial natriuretic peptide in neonatal rat cardiac fibroblasts: role in regulation of the local renin-angiotensin system.
There is an inverse relationship between renin and atrial natriuretic peptide (ANP) levels in the plasma. Since both the ANP and renin-angiotensin system (RAS) are upregulated in development and cardiac hypertrophy, we tested whether ANP differentially regulates RAS in cardiac cells. Cardiac fibroblasts isolated from neonatal rats were treated with ANP(1-28), a biologically active fragment of ANP. Renin and angiotensinogen (Ao) mRNA levels were measured by quantitative multiplex RT-PCR and protein levels determined by Western blot analysis. ANP(1-28) increased renin and Ao mRNA levels (737+/-131% and 178+/-51.3%) with EC50 values of 4.12+/-0.3 and 8.67+/-0.22 nmol/L, respectively. At the protein level, secretion of renin and Ao was significantly enhanced resulting in approximately 4-fold increase in ANG II level in the medium. The effect of ANP(1-28) on renin and Ao mRNA expression were reproduced by 8-bromo-cyclic GMP. Inhibition of protein kinase G (PKG) with KT5823 blunted ANP(1-28)-induced upregulation of renin, but not Ao mRNA, while inhibition of protein kinase A (PKA) with KT5720 attenuated the upregulation of both renin and Ao mRNA. These findings suggest that unlike in plasma, ANP positively regulates the RAS in cardiac fibroblasts, which may have a significant role in development of the fetal heart.


Striking similarity of the distribution patterns of the poly(ADP-ribose) polymerase and DNA polymerase beta among various mouse organs.
The expression level of poly(ADP-ribose) polymerase mRNA as well as the level of enzymatic activity were examined in various mouse organs by northern blot and activity gel analyses. High levels of the mRNA expression and enzymatic activity were observed in testis, thymus, spleen, and brain. On the other hand, low levels of the mRNA expression and enzymatic activity were observed in liver and kidney. These findings suggest that the expression of the poly(ADP-ribose) polymerase is mainly regulated by transcription. Striking similarity was observed between the patterns of organ distribution of enzymatic activities of poly(ADP-ribose) polymerase and DNA polymerase beta in various mouse organs.


The tryptophan hydroxylase activation inhibitor, AGN-2979, decreases regional 5-HT synthesis in the rat brain measured with alpha-[14C]methyl-L-tryptophan: an autoradiographic study.
Many experimental conditions are stressful for animals. It is well known that stress induces tryptophan hydroxylase (TPH) activation, resulting in increased serotonin (5-HT) synthesis. In our experimental procedure to measure 5-HT synthesis using alpha-[(14)C]methyl-L-tryptophan (alpha-MTrp) autoradiographic method, the hind limbs of animals are restrained using a loose-fitted plaster cast such that the forelimbs of the animal remain free. The objective of the present investigation was to evaluate the changes, if any, in 5-HT synthesis, after injecting these restrained rats with the TPH activation inhibitor AGN-2979. The effect on regional 5-HT synthesis was studied using the alpha-MTrp autoradiographic method. The hypothesis was that the TPH activation inhibitor would reduce 5-HT synthesis, if TPH activation was induced by this restraint. The rats received injection of AGN-2979 (10 mg/kg, i.p.) or distilled water vehicle (1 mL/kg, i.p.) 1 h prior to tracer administration. The free- and total tryptophan concentrations were not significantly different between the treatment and control groups. The results demonstrate that 5-HT synthesis in AGN-2979 treated rats is significantly decreased (-12 to -35%) in both the raphe nuclei and their terminal areas when compared to the control rats. These findings suggest that restrained conditions, such as those used in our experimental protocol, induce TPH activation resulting in an increased 5-HT synthesis throughout the brain. The reduction in 5-HT synthesis in the AGN-2979 group is not related to a change in the plasma tryptophan. Because there was no activation in the pineal body, the structure having a different isoform of TPH, we can propose that it is only the brain TPH that becomes activated with this specific restraint.


Monolithically Integrated μLEDs on Silicon Neural Probes for High-Resolution Optogenetic Studies in Behaving Animals.
We report a scalable method to monolithically integrate microscopic light emitting diodes (μLEDs) and recording sites onto silicon neural probes for optogenetic applications in neuroscience. Each μLED and recording site has dimensions similar to a pyramidal neuron soma, providing confined emission and electrophysiological recording of action potentials and local field activity. We fabricated and implanted the four-shank probes, each integrated with 12 μLEDs and 32 recording sites, into the CA1 pyramidal layer of anesthetized and freely moving mice. Spikes were robustly induced by 60 nW light power, and fast population oscillations were induced at the microwatt range. To demonstrate the spatiotemporal precision of parallel stimulation and recording, we achieved independent control of distinct cells ∼ 50 μm apart and of differential somato-dendritic compartments of single neurons. The scalability and spatiotemporal resolution of this monolithic optogenetic tool provides versatility and precision for cellular-level circuit analysis in deep structures of intact, freely moving animals.


Concealing Stuttering at School: "When You Can't Fix It…the Only Alternative Is to Hide It".
The purpose of this study was to explore experiences with concealing stuttering in children and young people who stutter based on recollections from adults. In addition, we explored how school-based speech therapists can be helpful or unhelpful to children who are concealing stuttering from the perspective of adults who stutter. Thirty adults who stutter, who previously or currently conceal stuttering, participated in semistructured interviews exploring their early experiences with hiding stuttering. Purposeful and random sampling was used to diversify experiences and opinions. Reflexive thematic analysis was used to develop themes and subthemes to describe participants' experiences. All participants in the study reported beginning to conceal stuttering at 18 years of age or younger, with more than two thirds sharing that they began in elementary school. Participants reported that exposure to implicit and explicit ableist messaging about stuttering and traumatic social experiences at school contributed to their inclination to hide disfluencies. Many participants described concealment as a strategy for protecting themselves from stigma. Several participants condemned fluency shaping, calling it harmful and likening it to teaching concealment. Participants believed that speech therapists could be helpful by promoting safe and supportive school environments and by being responsive to the social and emotional challenges that can accompany speaking differently and navigating stigma at school. Some children who stutter may attempt to protect themselves from stigma by concealing their disfluencies, but doing so can feel isolating and confusing. Speech therapists can play an important role in making the school environment safer and more supportive for children who stutter.