Human adaptation to immobilization: Novel insights of impacts on glucose disposal and fuel utilization.
Bed rest (BR) reduces whole-body insulin-stimulated glucose disposal (GD) and alters muscle fuel metabolism, but little is known about metabolic adaptation from acute to chronic BR nor the mechanisms involved, particularly when volunteers are maintained in energy balance. Healthy males (n = 10, 24.0 ± 1.3 years), maintained in energy balance, underwent 3-day BR (acute BR). A second cohort matched for sex and body mass index (n = 20, 34.2 ± 1.8 years) underwent 56-day BR (chronic BR). A hyperinsulinaemic euglycaemic clamp (60 mU/m2 /min) was performed to determine rates of whole-body insulin-stimulated GD before and after BR (normalized to lean body mass). Indirect calorimetry was performed before and during steady state of each clamp to calculate rates of whole-body fuel oxidation. Muscle biopsies were taken to determine muscle glycogen, metabolite and intramyocellular lipid (IMCL) contents, and the expression of 191 mRNA targets before and after BR. Two-way repeated measures analysis of variance was used to detect differences in endpoint measures. Acute BR reduced insulin-mediated GD (Pre 11.5 ± 0.7 vs. Post 9.3 ± 0.6 mg/kg/min, P < 0.001), which was unchanged in magnitude following chronic BR (Pre 10.2 ± 0.4 vs. Post 7.9 ± 0.3 mg/kg/min, P < 0.05). This reduction in GD was paralleled by the elimination of the 35% increase in insulin-stimulated muscle glycogen storage following both acute and chronic BR. Acute BR had no impact on insulin-stimulated carbohydrate (CHO; Pre 3.69 ± 0.39 vs. Post 4.34 ± 0.22 mg/kg/min) and lipid (Pre 1.13 ± 0.14 vs. Post 0.59 ± 0.11 mg/kg/min) oxidation, but chronic BR reduced CHO oxidation (Pre 3.34 ± 0.18 vs. Post 2.72 ± 0.13 mg/kg/min, P < 0.05) and blunted the magnitude of insulin-mediated inhibition of lipid oxidation (Pre 0.60 ± 0.07 vs. Post 0.85 ± 0.06 mg/kg/min, P < 0.05). Neither acute nor chronic BR increased muscle IMCL content. Plentiful mRNA abundance changes were detected following acute BR, which waned following chronic BR and reflected changes in fuel oxidation and muscle glycogen storage at this time point. Acute BR suppressed insulin-stimulated GD and storage, but the extent of this suppression increased no further in chronic BR. However, insulin-mediated inhibition of fat oxidation after chronic BR was less than acute BR and was accompanied by blunted CHO oxidation. The juxtaposition of these responses shows that the regulation of GD and storage can be dissociated from substrate oxidation. Additionally, the shift in substrate oxidation after chronic BR was not explained by IMCL accumulation but reflected by muscle mRNA and pyruvate dehydrogenase kinase 4 protein abundance changes, pointing to lack of muscle contraction per se as the primary signal for muscle adaptation.


Keloid morphea and nodular scleroderma: two distinct clinical variants of scleroderma?
For nearly a century, the terms "keloid morphea" and "nodular scleroderma" have been used interchangeably without defined clinical or histologic criteria. To define the conditions "keloid morphea" and "nodular scleroderma" by correlating the clinical and histologic features. We retrospectively identified six patients with keloidal lesions and nodules from 70 consecutive patients with scleroderma seen in the dermatology clinic. The clinical presentation and histopathological findings were reviewed. Six of 70 patients with scleroderma (45 systemic and 25 morphea) exhibited keloidal or nodular lesions. All these patients had systemic sclerosis. Clinically one patient (case 1) had nodules; five (cases 2-6) had keloids. The nodular lesions had histologic findings consistent with keloid, while the keloidal plaques were variably keloids or morphea histopathologically. There was no correlation between the clinical morphology and the histologic findings, except for cases 5 and 6. These patients were African-American, with a family history of keloids and typical keloids clinically and histologically that developed from sites with normal skin. Keloid morphea and nodular scleroderma are clinical terms that describe keloidal and nodular lesions in patients most commonly with scleroderma. The clinicopathologic association is variable. Based on a review of the English literature and our series of six patients, we identified two clinical variants; (1) keloidal or nodular lesions arising from sclerodermatous skin with histologic findings of keloid or scleroderma, (2) typical keloids clinically and histologically, arising in normal skin in patients with a family history of keloids. Awareness of these entities is important for proper diagnosis of the cutaneous lesions and for recognizing that the cutaneous findings may be a sign of systemic sclerosis


Biomonitoring air pollution with the desert lichen Ramalina maciformis.
To investigate the environmental impact of anthropogenic activity in the Negev Desert, we monitored the following in order to determine the physiological integrity of the epilithic fruticose lichen Ramalina maciformis: (1) production of ethylene; (2) potential quantum yield of photosystem II expressed as chlorophyll fluorescence ratio Fv/Fm; and (3) electrolyte leakage of cell membranes in terms of electric conductivity. The degree of pollution was assessed by measuring the amounts of B, Cd, Co, Cu, Fe, K, Mg, Mn, Na, Ni, P, Pb, Sr and Zn in the lichen thallus. Some of the lichen-carrying stones collected in the relatively unpolluted control site were relocated on the same hill, to test the possible impact of relocation. An additional amount of lichen-carrying stones was marked as controls. The greater part of the lichen-carrying stones was transferred to 10 biomonitoring sites in and around Ramat Hovav, an industrial area in the Negev Desert, Israel. Transplanted lichen in polluted sites contained higher amounts of mineral elements than lichens in the control site after an exposure period of 7 months. Statistical analysis revealed positive correlation of ethylene production and Ni content, negative correlation of Fv/Fm ratios and B, Cu, Mn, Na, Ni, Sr and Zn, and positive correlation of electric conductivity and B, Mn, Ni and Sr. Both elemental content and physiological alterations in transferred samples of R. maciformis point to a high degree of contamination in Ramat Hovav. The present study suggests that the lichen R. maciformis should be classified as a sensitive species endangered by anthropogenic activity in the desert. Furthermore, this lichen species is likely to be suitable as a bioindicator of pollution in other parts of the Negev Desert. Ethylene production and electric conductivity as parameters of lichen-vitality, indicated stress in 5 of the 12 biomonitoring sites (42%) and may therefore be considered as more sensitive than the Fv/Fm ratio, which indicated stress in 3 of the 12 sites (25%).


Temporal analysis of tissue displacement induced by a transient ultrasound radiation force.
One of the stress sources that can be used in dynamic elastography imaging methods is the acoustic radiation force. However, displacements of the medium induced by this stress field are generally not fully understood in terms of spatial distribution and temporal evolution. A model has been developed based on the elastodynamic Green's function describing the different acoustic waves generated by focused ultrasound. The function is composed of three terms: two far-field terms, which correspond to a purely longitudinal compression wave and a purely transverse shear wave, and a coupling near-field term which has a longitudinal component and a transverse component. For propagation distances in the shear wavelength range, the predominant term is the near field term. The displacement duration corresponds to the propagation duration of the shear wave between the farthest source point and the observation point. This time therefore depends on the source size and the local shear modulus of the tissue. Evolution of the displacement/time curve profile, which is directly linked to spatial and temporal source profiles, is computed at different radial distances, for different durations of force applications and different shear elastic coefficients. Experimental results performed with an optical interferometric method in a homogeneous tissue-mimicking phantom agreed with the theoretical profiles.


Poly(Lactic-co-Glycolic Acid): Applications and Future Prospects for Periodontal Tissue Regeneration.
Periodontal tissue regeneration is the ultimate goal of the treatment for periodontitis-affected teeth. The success of regenerative modalities relies heavily on the utilization of appropriate biomaterials with specific properties. Poly (lactic-co-glycolic acid) (PLGA), a synthetic aliphatic polyester, has been actively investigated for periodontal therapy due to its favorable mechanical properties, tunable degradation rates, and high biocompatibility. Despite the attractive characteristics, certain constraints associated with PLGA, in terms of its hydrophobicity and limited bioactivity, have led to the introduction of modification strategies that aimed to improve the biological performance of the polymer. Here, we summarize the features of the polymer and update views on progress of its applications as barrier membranes, bone grafts, and drug delivery carriers, which indicate that PLGA can be a good candidate material in the field of periodontal regenerative medicine.


Reinnervation of denervated muscle by transplantation of fetal spinal cord to transected sciatic nerve in the rat.
When motor neurons in the spinal cord are destroyed, regeneration of motor axons and muscle reinnervation cannot be expected. We attempted reinnervation of the denervated muscle, i.e. motor unit reconstruction, using transplantation of the fetal spinal cord to the peripheral nerve. The sciatic nerve of an adult rat was resected for 20 mm, and a cavity was prepared using an autologous femoral vein at the distal stump of the nerve. The fetal spinal cord was then transplanted into the venous cavity. After 3-6 months, no voluntary muscle contraction was observed due to the absence of communication with the central nervous system. However, reinnervation of the muscles via the sciatic nerve by the transplanted spinal neurons was demonstrated electrophysiologically and histochemically. This suggested that a motor unit can be reconstructed by fetal spinal cord transplantation even if the original motor neurons in the spinal cord are not available.


Hypoxia-induced autophagy: cell death or cell survival?
Hypoxia (approximately 3-0.1% oxygen) is capable of rapidly inducing, via the hypoxia-inducible factor (HIF-1), a cell survival response engaging autophagy. This process is mediated by the atypical BH3-only proteins the Bcl-2/E1B 19kDa-interacting protein 3 (BNIP3/BNIP3L (NIX)) that are induced by HIF-1. These mitochondrial associated BNIP proteins also mediate mitophagy, a metabolic adaptation for survival that is able to control reactive oxygen species (ROS) production and DNA damage. In contrast, severe hypoxic conditions or anoxia (<0.1% oxygen), where the latter is often confused with physiological hypoxia, are capable of inducing a HIF-independent autophagic response, generated via an extreme nutritional stress response implicating the AMPK-mTOR and unfolded protein response (UPR) pathways. The autophagic cell death that is often observed in these extreme stress conditions should be seen as the outcome of failed adaptation.


Synthesis and biological activity of a novel series of indole-derived PPARgamma agonists.
The synthesis and structure-activity relationships of a novel series of indole 5-carboxylic acids that bind and activate peroxisome proliferator-activated receptor gamma (PPARgamma) are reported. These new analogs are selective for PPARgamma vs the other PPAR subtypes, and the most potent compounds in this series are comparable to in vitro potencies at PPARgamma reported for the thiazolidinedione-based antidiabetic drugs currently in clinical use.


Multifunctional Tandem Peptide Modified Paclitaxel-Loaded Liposomes for the Treatment of Vasculogenic Mimicry and Cancer Stem Cells in Malignant Glioma.
The chemotherapy of aggressive glioma is usually accompanied by a poor prognosis because of the formation of vasculogenic mimicry (VM) and brain cancer stem cells (BCSCs). VM provided a transporting pathway for nutrients and blood to the extravascular regions of the tumor, and BCSCs were always related to drug resistance and the relapse of glioma. Thus, it is important to evaluate the inhibition effect of antiglioma drug delivery systems on both VM and BCSCs. In this study, paclitaxel-loaded liposomes modified with a multifunctional tandem peptide R8-c(RGD) (R8-c(RGD)-Lip) were used for the treatment of glioma. An in vitro cellular uptake study proved the strongest targeting ability to be that of R8-c(RGD)-Lip to glioma stem cells. Drug loaded R8-c(RGD)-Lip exhibited an efficient antiproliferation effect on BCSCs and could induce the destruction of VM channels in vitro. The following pharmacodynamics study demonstrated that R8-c(RGD)-modified drug-loaded liposomes achieved both anti-VM and anti-BCSC effects in vivo. Finally, no significant cytotoxicity of the blood system or major organs of the drug-loaded liposomes was observed under treatment dosage in the safety evaluation. In conclusion, all of the results proved that R8-c(RGD)-Lip was a safe and efficient antiglioma drug delivery system.


Real-Time Zeta Potential Analysis of Microchannel Surfaces during Aminosilane Deposition and Exposure Using Current Monitoring.
Surface coatings are extensively used in capillary electrophoresis to increase separation efficiency and resolution. The stability of these coatings across a wide pH range is desirable to achieve repeatable migration times; therefore, a comprehensive understanding of coating degradation timescales is needed. We present a novel platform for automated zeta potential analysis based upon current monitoring that delivers improved time resolution over the existing methods. Using our platform, we measure the zeta potential continuously during aminosilane coating reactions and infer changes in the surface composition. We found that the change in the zeta potential after coating depended on the monomer type and solvent, while its stability was influenced by the coating solvent and exposure pH. Our versatile platform provides an elegant approach for evaluating the molecular composition, reactivity, and stability of surfaces in real time.


Treatment of dysphagia after stroke with acupuncture and related therapies: A protocol for systematic review and network meta-analysis.
Dysphagia is a common complication after stroke, with high disability rate and high fatality rate. Although several clinical studies and evidence-based medicine have demonstrated the efficacy of acupuncture in the treatment of dysphagia after stroke, there are significant differences in study design and intervention methods. The objective of this study is to compare the efficacy and safety of different acupuncture and related therapies in the treatment of dysphagia after stroke, so as to provide a superior clinical program. We will search 7 databases for randomized controlled trials of acupuncture-related therapies for dysphagia after stroke, including PubMed, the Cochrane Library, EMbase, China National Knowledge Infrastructure, China Biological Medicine, Chinese Scientific Journals Database, and wan-fang databases, from the date of the establishment of each database to March 31, 2020. The network meta-analysis will be implemented through Aggregate Data Drug Information System 1.16.8 and Stata 13.0 software. Clinical Efficiency, videofluoroscopic swallowing study score and Kubota Drinking Water Test grade will be the primary outcomes, Swallowing disorder specific quality of life score, Standardized Assessment and Adverse effects will be evaluated as secondary outcomes. Mean differences or odds ratios will be used for statistical analysis. We will ensure the reliability of the results through node-split model and heterogeneity analysis. In addition, methodological quality will be evaluated based on the Cochrane Collaboration's tool, and the quality of evidence will be evaluated according to the Grading of Recommendations Assessment, Development and Evaluation system. This study will provide a reliable evidence for the selection of acupuncture and related therapies for dysphagia after stroke. The results of this study will provide references for evaluating the influence of acupuncture and related therapies for dysphagia after stroke, and provide decision-making references for clinical research. This study did not require ethical approval. We will disseminate our findings by publishing results in a peer-reviewed journal. DOI 10.17605/OSF.IO/TAHND.


[Evaluation of resistance to anti-retroviral agents. Publication of clinical trials].
Information on patterns of resistance to and cross-resistance between antiretroviral agents is increasingly available and may be important for decisions on how to combine drugs to achieve an optimum antiviral effect in future therapeutic strategies. The increasing number of heavily pre-treated patients have changed the approach to assist therapeutic decision-making in patients failing therapy. Recent studies give us a more comprehensive analysis of the link of such mutations with a rebound in viral load and appearance of drug resistance. However, not all sites of mutations are known and the effects of interactions between them at different codons has not yet been well understood. Moreover, the interpretation is made from generalisations based on drug testing in vitro and there are scarce clinical data available. This work tries to summarize the most important studies to date, in order to know the significance of drug resistance as well as their potential use in everyday clinical practice.


Quantitative and molecular analysis of noroviruses RNA in blood from children hospitalized for acute gastroenteritis in Belém, Brazil.
Noroviruses (NoVs) are a common cause of acute gastroenteritis (AGE) and until now, little is known about its ability to spread outside the gut. We aim to investigate the role of NoVs causing viremia in children hospitalized for AGE, as well as to correlate the presence of NoVs RNA in serum with clinical severity and stool viral load. Paired stool and serum samples were collected from 85 pediatric patients under 6 years hospitalized for AGE from March to September 2012 in Belém, Brazil. Enzyme-linked immunosorbent assay (EIA) and reverse transcription quantitative PCR (RT-qPCR) were used to detect and quantify NoVs, respectively. Phylogenetic analysis of the partial ORF2 region was used to genotype the strains detected. NoVs were detected in 34.1% (29/85) of stool samples. By qRT-PCR, we found a high rate of NoVs' RNA in serum samples (34.5%) among NoVs-positive AGE cases, and was associated with a longer hospitalization (6.5 vs. 4.0 days; p=0.006), as well as with a higher stool viral load (3.9×10(11) vs. 1.1×10(11) GC/g; p=0.0472). NoVs strains were classified as GII.4 (90% of genotyped strains) and GII.7 (10%). The same genotype was found in paired stool and serum samples. Detection and molecular characterization of NoVs GII in paired stool and serum samples suggest that the dissemination of NoVs to the blood stream is not uncommon, but the role of viruses spread outside the gut and the relationship with disease severity need to be further addressed.


jefA (Rv2459), a drug efflux gene in Mycobacterium tuberculosis confers resistance to isoniazid & ethambutol.
Drug efflux pumps have been contributing factor(s) in the development of multidrug resistance in various clinically relevant bacteria. During efflux pump gene expression studies on mycobacteria, we have found a previously uncharacterized open reading frame (ORF) Rv2459 to be overexpressed in drug stressed conditions. The objective of the present study was to investigate the role of this ORF as a drug efflux pump, which might add new information in our understanding about the alternative mechanisms of drug resistance in mycobacteria. The open reading frame Rv2459 of Mycobacterium tuberculosis encoding a probable drug efflux protein has been cloned using pSD5 E.coli-Mycobacterium shuttle vector and overexpressed in M. tuberculosis H(37)Rv. This ORF was named as jefA. Overexpression of this gene in clones has been verified by real-time reverse transcription PCR. Minimum inhibitory concentrations (MICs) of recombinant as well as non-recombinant clones were determined by resazurin microtitre assay plate method (REMA) with and without efflux pump inhibitors carbonyl cyanide m-chlorophenylhydrazone (CCCP) and verapamil. In recombinant strains of M. tuberculosis, the overexpression of this gene led to an increase in MIC of anti-tubercular drugs isoniazid and ethambutol when tested by REMA. In the presence of CCCP and verapamil, the recombinant strains showed decrease in MIC for these drugs. Bioinformatic analysis has shown a close relation of JefA protein with drug efflux pumps of other clinically relevant bacteria. In homology derived structure prepared from nearest available model, it was observed that amino acids forming TMH 1, 8 and 11 participated in ethambutol specificity and those forming TMH 2, 7 and 10 participated in isoniazid specificity in JefA. The increased transcription of jefA leads to increased resistance to ethambutol and isoniazid in M. tuberculosis via efflux pump like mechanism and contributes in the development of resistance to these drugs. JefA amino acid sequence is well conserved among clinically important bacterial genera, which further provides evidence of being a potent drug efflux pump. The involvement in drug resistance and very little homology with any of the human proteins makes JefA important to be included in the list of potential drug targets.


Low-loss hollow-core silica fibers with adjacent nested anti-resonant tubes.
We report on numerical design optimization of hollow-core anti-resonant fibers with the aim of reducing transmission losses. We show that re-arranging the nested anti-resonant tubes in the cladding to be adjacent has the effect of significantly reducing leakage as well as bending losses, and for reaching high loss extinction ratios between the fundamental mode and higher order modes. We investigate two versions of the proposed design, one optimized for the mid-IR and another scaled down version for the near-IR and compare them in detail with previously proposed anti-resonant fiber designs including nested elements. Our proposed design is superior with respect to obtaining the lowest leakage losses and the bend losses are also much lower than for the previous designs. Leakage losses as low as 0.0015 dB/km and bending losses of 0.006 dB/km at 5 cm bending radius are predicted at the ytterbium lasing wavelength 1.06 µm. When optimizing the higher-order-mode extinction ratio, the low leakage loss is sacrificed to get an effective single-mode behavior of the fiber. We show that the higher-order-mode extinction ratio is more than 1500 in the range 1.0-1.1 µm around the ytterbium lasing wavelength, while in the mid-IR it can be over 100 around λ = 2.94 μm. This is higher than the previously considered structures in the literature using nested tubes.


Neural field theory of nonlinear wave-wave and wave-neuron processes.
Systematic expansion of neural field theory equations in terms of nonlinear response functions is carried out to enable a wide variety of nonlinear wave-wave and wave-neuron processes to be treated systematically in systems involving multiple neural populations. The results are illustrated by analyzing second-harmonic generation, and they can also be applied to wave-wave coalescence, multiharmonic generation, facilitation, depression, refractoriness, and other nonlinear processes.


Cadmium tolerance and antibiotic resistance of Pseudomonas sp. isolated from water, sludge and fish raised in wastewater-fed tropical ponds.
The numbers of Pseudomonas sp. isolated were counted in samples collected from water, sludge and intestine of fishes raised in different wastewater ponds along an effluent gradient in a sewage treatment plant. Total fish yield in the last maturation pond increased by 73% over the yield in first maturation pond or facultative pond. The number of Pseudomonas sp. isolated from the intestine of the tilapia (Oreochromis mssambicus) raised in facultative pond, was more than three times the counts (7.22 x 10(8)/g) observed in the last maturation pond (2.025 x 10(8)/g). The effective lethal concentration of cadmium for Pseudomonas sp. isolated from the intestine of the tilapia was 0.6 mM and 0.08-0.09 mM when the fish was procured from facultative pond and last maturation pond, respectively. The Pseudomonas sp. isolated from the intestine of the tilapia did not have resistance to any of the ten antibiotics tested. However, the bacterium isolated from raw sewage, water and sediment of the anaerobic pond was resistant to seven out of ten antibiotics tested.


Natural Loss of eyeless/Pax6 Expression in Eyes of Bicyclus anynana Adult Butterflies Likely Leads to Exponential Decrease of Eye Fluorescence in Transgenics.
Commonly used visible markers for transgenesis use fluorescent proteins expressed at the surface of the body, such as in eyes. One commonly used marker is the 3xP3-EGFP cassette containing synthetic binding sites for the eyeless/Pax6 conserved transcription factor. This marker cassette leads to fluorescent eyes in a variety of animals tested so far. Here we show that upon reaching adulthood, transgenic Bicyclus anynana butterflies containing this marker cassette exponentially loose fluorescence in their eyes. After 12 days, transgenic individuals are no longer distinguishable from wild type individuals. The decreased eye fluorescence is likely due to significantly decreased or halted eyeless/Pax6 expression observed in wild type animals upon adult emergence. Implications from these findings include care in screening transgenic animals before these reach adulthood, or shortly thereafter, and in using adult animals of the same age for quantitative screening of likely homozygote and heterozygote individuals.


Direct growth of polyaniline chains from N-doped sites of carbon nanotubes.
Polymer grafting from graphitic carbon materials has been pursued for several decades. Unfortunately, currently available methods mostly rely on the harsh chemical treatment of graphitic carbons which causes severe degradation of chemical structure and material properties. A straightforward growth of polyaniline chain from the nitrogen (N)-doped sites of carbon nanotubes (CNTs) is presented. N-doping sites along the CNT wall nucleate the polymerization of aniline, which generates seamless hybrids consisting of polyaniline directly grafted onto the CNT walls. The resultant materials exhibit excellent synergistic electrochemical performance, and can be employed for charge collectors of supercapacitors. This approach introduces an efficient route to hybrid systems consisting of conducting polymers directly grafted from graphitic dopant sites.


Why in the world would anyone want to work for you?
Why in the world would someone want to work or keep working at your facility? If you can't answer this question, you will have a very difficult time finding and keeping the people you need to grow your agency. Home care managers agree that keeping good people is a top strategy. What is needed are clear strategies, concrete action plans, and methods to measure the results.


Imaging Natural Occlusal Caries Lesions with Optical Coherence Tomography.
Several studies have demonstrated that polarization-sensitive optical coherence tomography (PS-OCT) can be used to nondestructively measure the severity of demineralization in the important occlusal surfaces. The purpose of this study was to assess the potential of PS-OCT and OCT methods for the measurement of the depth of natural occlusal carious lesions. Teeth were screened for potential occlusal lesions using near infrared imaging (NIR). A PS-OCT system operating at 1310-nm was used to acquire polarization resolved images of the area of interest on the occlusal surface. The teeth were serial sectioned to 200 μm thickness and examined with polarized light microscopy (PLM) and Transverse Microradiography (TMR) for comparison. The lesion depth measured nondestructively with PS-OCT was compared to the lesion depth measured with PLM and TMR to assess the performance of these methods and determine if polarization sensitivity is required. The lesion depth measured using OCT correlated well with the lesion depths measured with TMR and PLM. Although polarization sensitivity provided better contrast it was not necessary to have polarization sensitivity to identify deep occlusal lesions.


Biochemistry of Fern Spore Germination: Globulin Storage Proteins in Matteuccia struthiopteris L.
Two globulin storage proteins have been identified in spores of the ostrich fern, Matteuccia struthiopteris (L.) Todaro. The two proteins comprise a significant amount of the total spore protein, are predominantly salt-soluble, and can be extracted by other solvents to a limited extent. The large 11.3 Svedberg unit (S) globulin is composed of five polypeptides with molecular weights of 21,000, 22,000, 24,000, 28,000 and 30,000. Each polypeptide has several isoelectric point (pI) variants between pH 5 and 7. The small 2.2S storage protein has a pI > 10.5 and is composed of at least two major polypeptides of 6,000 and 14,000 M(r). The amino acid composition of both storage proteins reveals that the 11.3S protein is particularly rich in aspartic and glutamic acid, while the 2.2S protein has few acidic amino acids. During imbibition and germination the globulin fraction declines rapidly, with a corresponding degradation of individual polypeptides of each protein. Polyclonal antibodies against each of the two proteins were produced and used for immunolocalization to determine the site of storage protein deposition within the quiescent spore. The proteins were sequestered in protein bodies of 2 to 10 micrometers, that are morphologically similar to those found in the seeds of flowering plants. The results suggest that spore globulins are biochemically similar to seed globulins, especially those found in some cruciferous seeds.


Basic features of bovine spermatogonial culture and effects of glial cell line-derived neurotrophic factor.
Spermatogonial stem cells (SSC) are a small self-renewing subpopulation of type A spermatogonia, which for the rest are composed of differentiating cells with a very similar morphology. We studied the development of primary co-cultures of prepubertal bovine Sertoli cells and A spermatogonia and the effect of glial cell line-derived neurotropic factor (GDNF) on the numbers and types of spermatogonia, the formation of spermatogonial colonies and the capacity of the cultured SSC to colonize a recipient mouse testis. During the first week of culture many, probably differentiating, A spermatogonia entered apoptosis while others formed pairs and chains of A spermatogonia. After 1 week colonies started to appear that increased in size with time. Numbers of single (A(s)) and paired (A(pr)) spermatogonia were significantly higher in GDNF treated cultures at Days 15 and 25 (P < 0.01 and 0.05, respectively), and the ratio of A(s) to A(pr) and spermatogonial chains (A(al)) was also higher indicating enhanced self-renewal of the SSC. Furthermore, spermatogonial outgrowths in the periphery of the colonies showed a significantly higher number of A spermatogonia with a more primitive morphology under the influence of GDNF (P < 0.05). Spermatogonial stem cell transplantation experiments revealed a 2-fold increase in stem cell activity in GDNF treated spermatogonial cultures (P < 0.01). We conclude that GDNF rather than inducing proliferation, enhances self-renewal and increases survival rates of SSC in the bovine spermatogonial culture system.


Physiologic motion after vocal cord reinnervation: a preliminary study.
This study attempted to reestablish physiologic vocal cord motion, rather than synkinesis, to a reinnervated vocal cord. One mongrel dog underwent a division and reanastomosis of the anterior branch of the right recurrent laryngeal nerve and simultaneous separation and reimplantation of a posterior division nerve-muscle pedicle into the posterior cricoarytenoid muscle. After 21 weeks, spontaneous physiologic vocal cord movement and electromyographic (EMG) activity were recorded during respiratory obstruction and laryngeal mechanical stimulation. Acoustic measures and histologic data are also presented from the reinnervated and normal vocalis muscle and from the recurrent laryngeal nerve. This study demonstrated that physiologic vocal cord motion can be achieved after laryngeal reinnervation using this technique.


A new level of architectural complexity in the human pyruvate dehydrogenase complex.
Mammalian pyruvate dehydrogenase multienzyme complex (PDC) is a key metabolic assembly comprising a 60-meric pentagonal dodecahedral E2 (dihydrolipoamide acetyltransferase) core attached to which are 30 pyruvate decarboxylase E1 heterotetramers and 6 dihydrolipoamide dehydrogenase E3 homodimers at maximal occupancy. Stable E3 integration is mediated by an accessory E3-binding protein (E3BP) located on each of the 12 E2 icosahedral faces. Here, we present evidence for a novel subunit organization in which E3 and E3BP form subcomplexes with a 1:2 stoichiometry implying the existence of a network of E3 "cross-bridges" linking pairs of E3BPs across the surface of the E2 core assembly. We have also determined a low resolution structure for a truncated E3BP/E3 subcomplex using small angle x-ray scattering showing one of the E3BP lipoyl domains docked into the E3 active site. This new level of architectural complexity in mammalian PDC contrasts with the recently published crystal structure of human E3 complexed with its cognate subunit binding domain and provides important new insights into subunit organization, its catalytic mechanism and regulation by the intrinsic PDC kinase.


In-vivo metabolism in the rat and mouse of antrafenine to 1-m-trifluoromethylphenylpiperazine.
The analgesic antrafenine forms 1-m-trifluoromethylphenylpiperazine (mCF3PP) during its biotransformation in the rat and mouse. At least 14 and 3% of an antrafenine dose (25 mg kg-1 p.o.) reaches the systemic circulation as mCF3PP in the mouse and rat respectively. The metabolite easily enters the brain, reaching concentrations several times those in body fluids. This, together with the fact that mCF3PP is known to produce several pharmacological effects compatible with a stimulatory action on 5-hydroxytryptamine postsynaptic receptors, suggests that this metabolite may contribute to the parent drug's pharmacological effects.


DNA damage in round spermatids of mice with a targeted disruption of the Pp1cgamma gene and in testicular biopsies of patients with non-obstructive azoospermia.
Non-obstructive azoospermia accounts for a considerable proportion of male factor infertility. Current therapies for treatment of this kind of infertility include procedures such as intracytoplasmic sperm injection (ICSI), round spermatid injection (ROSI), round spermatid nucleus injection (ROSNI) and elongated spermatid injection (ELSI). All involve injection of haploid germ cells retrieved from testicular biopsies into recipient oocytes. We have investigated a mouse model of azoospermia for quality of haploid germ cell genomes, based on 4,6-diamidino-2-phenylindole (DAPI)/TdT-mediated dUTP nick-end labelling (TUNEL) labelling. The mouse model, a targeted mutation in the protein phosphatase 1cg gene, results in severe depletion of haploid germ cells from the round spermatid stage on. Mice homozygous for the mutation are completely infertile, and produce only the occasional spermatozoon. Spermatozoa and round spermatids retrieved from either the epididymides or the testes of mutant mice displayed very high rates of DNA fragmentation. In contrast, similar cells retrieved from heterozygous or wild-type littermates displayed low levels of DNA fragmentation. In some cases, the high rates of DNA fragmentation in mutant cells could be lowered by inclusion of antioxidants in the retrieval media. High rates of DNA fragmentation were also observed in round spermatids retrieved from testicular biospies of human patients with non-obstructive azoospermia. These results suggest that one of the features of the pathology associated with azoospermia is fragmented DNA in haploid germ cells. This raises questions about the suitability of using these cells for fertility treatment.


Cat8p, the activator of gluconeogenic genes in Saccharomyces cerevisiae, regulates carbon source-dependent expression of NADP-dependent cytosolic isocitrate dehydrogenase (Idp2p) and lactate permease (Jen1p).
The yeast transcriptional activator Cat8p has been identified as a factor that is essential for the derepression of genes involved in gluconeogenesis (like FBP1, PCK1, ACR1, ICL1 and MLS1) when only nonfermentable carbon sources are provided. Cat8p-dependent expression is mediated by cis-acting elements in the respective promoters, which are named UAS/CSREs (upstream activating sequence/carbon source responsive element). To establish whether the function of Cat8p is restricted to the activation of gluconeogenesis or is also involved in the regulation of a greater variety of genes, we investigated the transcriptional regulation of two genes, IDP2 and JEN1, which exhibit a similar expression pattern to gluconeogenic genes, although IDP2 at least is not linked directly to the gluconeogenic pathway. We identified functional UAS/CSRE elements in the promoters of both genes. Expression studies revealed that JEN1 is regulated negatively by the repressors Mig1p and Mig2p, and that Cat8p is needed for full derepression of the gene under non-fermentative growth conditions. Furthermore, we showed that Mig2p is also involved in the repression of CAT8 itself. The results presented in this study support a model in which Cat8p-dependent gene activation is not restricted to gluconeogenesis, but targets a wide variety of genes which are strongly derepressed under non-fermentative growth conditions.


Clinical testing of thermally stimulated cardiovascular oscillations in man.
The study assessed the physiological validity of an automatic thermal stimulation method to induce synchronised oscillations in the neural cardiovascular control system. Automatic alternating rhythmic warm and cool immersion of different skin areas of 18 males was done at different frequencies and water temperatures. The neurally mediated responses to the periodic thermal stimulation were measured from skin blood flow and heart rate and compared to those of a sham stimulation. Respiration was monitored for control purposes. The reproducibility of the stimulation and responses was examined. 18 young males volunteered for the study. The water bath method produced reproducible thermal stimulation and responses of skin blood flow and heart rate. Rhythmic thermal stimulation at 0.013-0.096 Hz synchronised the oscillations of the forearm skin blood flow when the thermal stimulus amplitude exceeded 10 degrees C. The increase in the stimulus amplitude or enlargement of the stimulus area did not further increase the oscillatory response of skin blood flow. Sham stimulation or mean temperature of the periodic thermal stimulation in the range 23-33 degrees C did not influence the oscillations of skin blood flow. Local cooling of the stimulated lower legs attenuated the response of skin blood flow. Both thermal stimulation and sham stimulation affected heart rate, but no stable synchronisation of the periodic heart rate variability was found at supine rest. Thermal stimulation of the sitting subjects' forearm instead of legs increased the synchronisation of the periodic heart rate variability. The response of skin blood flow agreed with the theory of the thermal entrainment. In a supine man, both thermal stimulation and non-specific central nervous influences induced significant and reproducible interactions with periodic heart rate variability and respiration.


Saccadic eye movement metrics reflect surgical residents' fatigue.
Little is known about the effects of surgical residents' fatigue on patient safety. We monitored surgical residents' fatigue levels during their call day using (1) eye movement metrics, (2) objective measures of laparoscopic surgical performance, and (3) subjective reports based on standardized questionnaires. Prior attempts to investigate the effects of fatigue on surgical performance have suffered from methodological limitations, including inconsistent definitions and lack of objective measures of fatigue, and nonstandardized measures of surgical performance. Recent research has shown that fatigue can affect the characteristics of saccadic (fast ballistic) eye movements in nonsurgical scenarios. Here we asked whether fatigue induced by time-on-duty (~24 hours) might affect saccadic metrics in surgical residents. Because saccadic velocity is not under voluntary control, a fatigue index based on saccadic velocity has the potential to provide an accurate and unbiased measure of the resident's fatigue level. We measured the eye movements of members of the general surgery resident team at St. Joseph's Hospital and Medical Center (Phoenix, AZ) (6 males and 6 females), using a head-mounted video eye tracker (similar configuration to a surgical headlight), during the performance of 3 tasks: 2 simulated laparoscopic surgery tasks (peg transfer and precision cutting) and a guided saccade task, before and after their call day. Residents rated their perceived fatigue level every 3 hours throughout their 24-hour shift, using a standardized scale. Time-on-duty decreased saccadic velocity and increased subjective fatigue but did not affect laparoscopic performance. These results support the hypothesis that saccadic indices reflect graded changes in fatigue. They also indicate that fatigue due to prolonged time-on-duty does not result necessarily in medical error, highlighting the complicated relationship among continuity of care, patient safety, and fatigued providers. Our data show, for the first time, that saccadic velocity is a reliable indicator of the subjective fatigue of health care professionals during prolonged time-on-duty. These findings have potential impacts for the development of neuroergonomic tools to detect fatigue among health professionals and in the specifications of future guidelines regarding residents' duty hours.


A model of growth and migration.
"This paper contains a model with which the desirability of persistent urbanization can be examined. There are two regions: city and countryside. Joint restrictions on the technologies in each region and on preferences over the goods produced in each region are the driving force of migration. In equilibrium there is persistent migration from the countryside to the city. Even though there are externalities in production, the competitive equilibrium is Pareto optimal." (SUMMARY IN FRE)


Intravitreal Phacoemulsification Using Torsional Handpiece for Retained Lens Fragments.
To evaluate the results of intravitreal phacoemulsification with torsional hand piece in eyes with posteriorly dislocated lens fragments. In this prospective, interventional case series, 15 eyes with retained lens fragments following phacoemulsification were included. All patients underwent standard three-port pars plana vitrectomy and intravitreal phacoemulsification using sleeveless, torsional hand piece (OZiL™, Alcon's Infiniti Vision System). Patients were followed up for a minimum of six months to evaluate the visual outcomes and complications. The preoperative best-corrected visual acuity (BCVA) ranged from light perception to 0.3. No complications such as thermal burns of the scleral wound, retinal damage due to flying lens fragments, or difficult lens aspiration occurred during intravitreal phacoemulsification. Mean post-operative BCVA at the final follow-up was 0.5. Two eyes developed cystoid macular edema, which was managed medically. No retinal detachment was noted. Intravitreal phacoemulsification using torsional hand piece is a safe and effective alternative to conventional longitudinal phacofragmentation.


Effects of cyclization on stability, structure, and activity of α-conotoxin RgIA at the α9α10 nicotinic acetylcholine receptor and GABA(B) receptor.
α-Conotoxin RgIA is of interest as a lead in the development of drugs for neuropathic pain. It modulates the α9α10 nicotinic acetylcholine receptor (nAChR) and the GABA(B) receptor, both of which are implicated in antinociception. However, because of its peptidic nature, RgIA is potentially susceptible to generic problems encountered by peptide-based drugs of poor oral bioavailability, short biological half-life, and low stability. Here, we improved the biopharmaceutical properties of RgIA by backbone cyclization using 3-7 residue peptidic linkers. Cyclization with a six-residue linker does not perturb the overall structure of RgIA, improves selectivity for the GABA(B) receptor over the α9α10 nAChR, and improves stability in human serum. The results provide insights to further improve the therapeutic properties of RgIA and other conotoxins being considered as drug leads and confirm that cyclization is a readily applicable strategy to improve the stability of peptides with proximate N- and C-termini.


Law enforcement personnel are willing to change, but report influencing beliefs and barriers to optimised dietary intake.
Law enforcement personnel have been recognized as having a high risk for several lifestyle-related health conditions which, in combination with the nature of their work (sedentary roles interspersed with intermittent high-intensity activity, shift work, and a high stress-load), can have a negative impact on their health. The aim of this study was to investigate the dietary habits and factors or barriers influencing these habits within a cohort of law enforcement personnel in the United States of America. Cross-sectional data were obtained via validated paper-based surveys being the Perceived Barriers to Healthy Eating, Food Choice Questionnaire and Rapid Eating Assessment for Participants, Short Version. A total of 159 participants (median age = 27 [range 19-60] years; 74% males) participated. Barriers to healthy eating included being busy and irregular working hours. Overall, 91% (n = 143) placed high importance on consuming nutritious food and 80% (n = 126) on food high in vitamins and minerals. A further 80% (n = 127) emphasized high protein content and 41% (n = 62) followed a high protein diet. Barriers to healthy eating included busy lifestyle (60%, n = 94), and irregular working hours (41%, n = 64). Overall, 80% (n = 127) were very willing to make changes in eating habits to be healthier. Law enforcement officers know what they should eat and report convenience and health the most important factors guiding their food choices. Knowing this, officers find challenges putting good dietary practices into practice due to factors like a busy lifestyle and irregular work hours. Reportedly "very willing" to make changes in their eating habits to be healthier, future interventions should focus on how to effect changes to their eating habits as opposed to focussing on what to eat.


A Tailored Lipid Supplement Restored Membrane Fatty Acid Composition and Ameliorates In Vitro Biological Features of Human Amniotic Epithelial Cells.
Cell culture conditions influence several biological and biochemical features of stem cells (SCs), including the membrane lipid profile, thus limiting the use of SCs for cell therapy approaches. The present study aims to investigate whether the in vitro culture may alter the membrane fatty acid signature of human Amniotic Epithelial Cells (hAECs). The analysis of the membrane fatty acid composition of hAECs cultured in basal medium showed a loss in polyunsaturated fatty acids (PUFA), in particular in omega-6 (ω-6) content, compared to freshly isolated hAECs. The addition to the basal culture medium of a chemically defined and animal-free tailored lipid supplement, namely Refeed®, partially restored the membrane fatty acid signature of hAECs. Although the amelioration of the membrane composition did not prolong hAECs culture lifespan, Refeed® influenced cell morphology, counteracted the onset of senescence, and increased the migratory capacity as well as the ability of hAECs to inhibit Peripheral Blood Mononuclear Cell (PBMC) proliferation. This study provides new information on hAEC features during culture passages and demonstrates that the maintenance of the membrane fatty acid signature preserved higher cell quality during in vitro expansion, suggesting the use of lipid supplementation for SC expansion in cell-based therapies.


In utero manipulation of coat color formation by a monoclonal anti-c-kit antibody: two distinct waves of c-kit-dependency during melanocyte development.
Previous studies on mice bearing various mutations within the c-kit gene, dominant white spotting (W), indicate the functional role of this tyrosine kinase receptor in the development of melanocytes, germ cells and hematopoietic cells. Despite the availability of mice defective in the c-kit gene and a respectable understanding of the molecular nature of c-kit, however, it is not clear at what stage of gestation c-kit is functionally required for the development of each of these cell lineages. To address this question, we have used a monoclonal anti-c-kit antibody, ACK2, as an antagonistic blocker of c-kit function to interfere with the development of melanocytes during embryonic and postnatal life. ACK2 injected intradermally into pregnant mice entered the embryos where it blocked the proper development of melanocytes. This inhibitory effect was manifested as coat color alteration in the offspring. Furthermore, ACK2 injection also altered the coat color of neonatal and adult mice. Based on the coat color patterns produced by ACK2 administration at various stages before or after birth, the following conclusions are drawn: (i) during mid-gestation, c-kit is functionally required during a restricted period around day 14.5 post-coitum when a sequence of events leading to melanocyte entry into the epidermal layer occurs; (ii) during postnatal life, c-kit is required for melanocyte activation which occurs concomitantly with the hair cycle which continues throughout life after neonatal development of the first hair.


Antidiuretic effects of oxotremorine microinjected into the hypothalamic supraoptic and paraventricular nuclei in a water-loaded and ethanol-anesthetized rat.
The effect of oxotremorine, a muscarinic agonist, on urine outflow compared with the effects of other cholinergic agonists and inhibitory effects of cholinergic antagonists upon the cholinergic actions were studied by microinjecting drugs stereotaxically, unilaterally into the supraoptic (SON) or paraventricular nuclei (PVN) in the hypothalamus of the rat which was loaded with water and anesthetized with ethanol. Oxotremorine decreased the urine outflow in dose- and time-dependent manners when microinjected into these nuclei. The median effective doses (ED50) were approx. 0.3 and 0.2 nmol in SON an PVN, respectively, being much less than ED50 values for nicotine. The time course of the antidiuretic effects was relatively slow, with the minimal urine outflow at approx. 30 min and the duration of one or longer hours. The antidiuretic effects of oxotremorine in these nuclei as well as the effects of acetylcholine and nicotine were completely blocked by pretreatment with atropine. The pretreatment with hexamethonium inhibited partially the effects of nicotine, but was unable to inhibit the effects of oxotremorine and acetylcholine. The data suggest that the antidiuretic effects of cholinergic agonists in SON and PVN are predominantly mediated through a muscarinic type of acetylcholine receptor. A possible mechanism for the antidiuretic effects is discussed.


[Successful 24-hour pulmonary preservation in the use of hyperbaric oxygen].
24-hour lung preservation with modified Euro-Collins (E-C) solution under hyperbaric oxygen (OHP) was studied in the canine single lung transplantation model. A prostaglandin I2 (PGI2) was administered before cardiac arrest by St. Thomas solution. Lungs were flushed with modified E-C solution, then heart and lungs were excised en-bloc and immersed in 4 degrees C modified E-C solution for 24 hours in control group (N = 5). In OHP group (N = 5), heart and lung block were preserved similarly in the control group but placed in hyperbaric chamber (mixed gas 95% O2, 5% CO2) under 2 atmospheres absolute pressure. In the hyperbaric chamber, the lungs were expanded continuously by pressure difference between the inside and the outside of the lung. The left lung was transplanted into recipient dog and reperfused for 3 hours. Pulmonary function was assessed serially by measuring oxygen and carbon dioxide tensions in arterial blood and pulmonary vascular resistance by clamping the right pulmonary artery for 5 minutes. ATP before and after preservation was measured. In the control group, PaO2, after 24 hours preservation followed by reperfusion, was 177 +/- 88 mmHg, but decreased to 75 +/- 34 mmHg at 3 hours of reperfusion and bloody foamy sputum appeared in 4 dogs. Alternatively, PaO2 was 256 +/- 58 mmHg at 3 hours of reperfusion and foamy sputum did not appear in OHP group. There was a significant difference between the 2 groups (p < 0.05). PaCO2 at 3 hours of reperfusion was 41 +/- 3 mmHg in the control group, and 31 +/- 2 mmHg in the OHP group (p < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)


Diagnosis and treatment of endometriosis in a hooded capuchin (Sapajus apella).
Spontaneous endometriosis is common in women, great apes and Old World monkeys, but rare in New World monkeys. We report on the clinical diagnosis and successful combination of surgical and medical treatment in a case of spontaneous endometriosis in a diabetic hooded capuchin. Addressing this painful condition had a positive welfare impact.


Sugar accumulation in sugarcane: role of cell walls in sucrose transport.
A new method was devised to measure the concentration of sucrose in the free space of sugarcane stalks. The free space includes the aqueous phase of cell walls. The method differs substantially, in principle, from one used previously, but it gave the same result, namely, that sucrose concentrations in the free space can approach the level in the bulk of the tissue.


Advanced research on dopamine signaling to develop drugs for the treatment of mental disorders: Ser311Cys polymorphisms of the dopamine D2-receptor gene and schizophrenia.
Schizophrenia is a debilitating and complex mental disorder with a prevalence of approximately 1% worldwide. The etiology remains unclear, despite massive research efforts. Hyperactive dopaminergic signal transduction in the central nervous system is suggested to be involved in the pathophysiology of schizophrenia (the dopamine hypothesis). The dopamine D(2)-receptor (DRD2) gene is thus a promising candidate for associations with risk of schizophrenia. We investigated DRD2 and found a novel missense nucleotide change causing an amino acid substitution of serine with cysteine at codon 311 (Ser311Cys). We performed an association study using 156 schizophrenia patients and 300 controls. Cys311 in DRD2 was significantly associated with schizophrenia. Patients with the Cys311 allele displayed shorter duration of hospitalization and less severe negative symptoms and were more frequently married compared to patients without this allele, suggesting good response to treatment. We expanded samples to 291 patients with schizophrenia (including 11 postmortem brain samples), 579 controls, and 78 patients with affective disorders in a further case-control study. Cys311 was associated with schizophrenia, particularly in patients without negative symptoms, and bipolar disorder with mood-incongruent psychotic symptoms. Three meta-analyses using over 20 published studies confirmed the association. In vitro studies showed that Cys311-type D(2) receptor impairs dopamine-induced sequestration, which appears to be consistent with the dopamine hypothesis.


Hyperglycemia and hypoxia are interrelated in their teratogenic mechanism: studies on cultured rat embryos.
Hyperglycemia and hypoxia are well-known teratogens that may affect many animal species, including man. One of the main mechanisms of teratogenic action of both seems to be increased oxidative stress. The purpose of this study was to evaluate the hypothesis that in the developing embryo hyperglycemia also leads to hypoxia, both resulting in oxidative damage. The study was performed on 10.5-day-old rat embryos of the regular Sabra strain cultured for 28 hours in hyperglycemic or hypoxic conditions. Embryonic growth and rate of anomalies was assessed at the end of the culture period. The embryonic oxidative damage was investigated by studying the levels of Malondialdehyde (MDA) to determine the lipid peroxidation. The redox status was studied by measuring the activity of Catalase-like (CAT) and Super Oxide Dismutase (SOD) enzymes and the amount of Low Molecular Weight Antioxidants (LMWA). In addition, we studied by immunohistochemistry in the embryos and yolk sacs the amount of nitrotyrosine as an additional marker for the extent of oxidative stress. The amounts of the redox and hypoxia sensitive transcription factors HIF1alpha, NFkB, and IkB were also studied by immunohistochemistry. Both hyperglycemia and hypoxia increased the rate of congenital anomalies mainly of the heart, neural tube, and brain. Embryonic growth and scoring were decreased only by hypoxia. Both hyperglycemia and hypoxia increased embryonic oxidative stress as evidenced by increased lipid peroxidation, increased nitrotyrosine and LMWA, but only minimal changes in CAT and SOD activity. Severe hyperglycemia also caused hypoxia, as evidenced by increased HIF1alpha. Thus, there seems to be an interrelation between hyperglycemia and hypoxia, both resulting in embryonic damage apparently by enhanced oxidative stress. Both hyperglycemia and hypoxia seem to exert their embryotoxicity through a similar mechanism of increased oxidative stress in the embryo in a stage when its antioxidant capacity is still weak. Moreover, hyperglycemia also seems to induce hypoxia, intensifying diabetes-induced embryopathy.


Immunogold localization of photosynthetic fructose-1,6-bisphosphatase in pea leaf tissue.
An enriched IgG serum fraction obtained from rabbits immunized against pea chloroplast fructose-1,6-bisphosphatase (FBPase) was used, coupled to colloidal gold (15 nanometer particles) goat anti-rabbit IgG, to analyze by electron microscopy the location of photosynthetic FBPase in pea (Pisum sativum L.) leaf ultrathin sections. In accordance with earlier biochemical studies on distribution of FBPase activity, the enzyme was visualized both in the stromal space and bound to the chloroplast membranes. Some gold particles also appear in the cytoplasm, which can be related to the presence in the cytosol of a high molecular weight precursor of this nuclear coded enzyme.


Lower striatal dopamine D2/3 receptor availability in obese compared with non-obese subjects.
Obesity is a result of a relative excess in energy intake over energy expenditure. These processes are controlled by genetic, environmental, psychological and biological factors. One of the factors involved in the regulation of food intake and satiety is dopaminergic signalling. A small number of studies have reported that striatal dopamine D2/D3 receptor [D2/3R] availability is lower in morbidly obese subjects. To confirm the role of D2/3R in obesity, we measured striatal D2/3R availability, using [123I]IBZM SPECT, in 15 obese women and 15 non-obese controls. Striatal D2/3R availability was 23% (p = 0.028) lower in obese compared with non-obese women. This study is an independent replication of the finding that severely obese subjects have lower striatal D2/3R availability. Our findings invigorate the evidence for lower striatal D2/3R availability in obesity and confirm the role of the striatal dopaminergic reward system in obesity.


Spectrum of inherited bleeding disorders from Western India.
The study was undertaken to assess the magnitude and diversity of different bleeding disorders in Western India. 768 cases referred to our Institute for evaluation of an underlying bleeding diathesis were investigated appropriately to detect the cause of the abnormal hemostatic function. 630 patients were diagnosed to have hereditary bleeding diathesis. Amongst these, 598 patients had a coagulation disorder while only 32 patients had a platelet function abnormality. Amongst the coagulation disorders, hemophilia A (70.5%) was the most common disorder followed by hemophilia B (14%) and VWD (10.8%). Glanzman's thrombasthenia (84.3%) was the most common platelet function disorder followed by Bernard-Soulier syndrome (12.5%). Some rare disorders have also been diagnosed. In spite of their apparent rarity, India has a substantial number of cases of inherited bleeding disorders. A large number of these patients is referred to many tertiary care institutions. It is therefore desirable that district hospitals must develop their laboratories to detect most of these disorders so that the patients need not travel long distances to get an appropriate diagnosis and proper management. All 1st degree female relatives of severe and moderate hemophilia must get factor assays done because some of them may be vulnerable to post-procedural or post-traumatic bleeding.


[Gout - a common and well known disease].
Hyperuricemia is defined by a blood urate level > 405 µmol/L, the cut-off value at which urate forms crystals in vivo. In 15-20% these individuals develop gout, clinically characterized by attacks of acute arthritis, initially and most commonly affecting MTP 1 or other joints, tendons and soft tissues of the foot. These attacks usually subside within 1 to 2 weeks. Over time attacks occur more frequently and can transform into chronic arthritis characterized by tophi. The gold standard for diagnosis relies on identification of urate crystals by polarization microscopy in aspirated joint fluid. This procedure is rarely performed in primary care where the majority of patients are seen, and gout is usually diagnosed by clinical criteria. New imaging technologies (ultrasound, dual-energy CT) can be helpful when aspiration is not available and when the diagnosis is unclear. Gout has a prevalence of 1.7% and incidence rate of approximately 200 per 100000 person-years in Sweden, figures that increase over time.


[Halogen-induced panniculitis caused by potassium bromide].
Necrotizing panniculitis due to potassium-bromide is a drug induced allergic reaction following stimulation of lymphocytes as demonstrated for the first time by a lymphocyte transformation test (LTT). We named the disease "halogen panniculitis" because of similar generally known reactions to iodides and describe the typical symptoms in three own cases. Bromoderma tuberosum tends to be a similar kind of allergy. For the first time pancreatitis to potassium bromide in men has been observed, which has already been described in epileptic dogs treated with potassium bromide.


Mandibular degloving injury in an Arabian filly.
A 6-month-old Arabian filly escaped its handler while being led and slipped on pavement. The referring veterinarian recognized severe, soft tissue damage to the filly's lower jaw and referred the filly to the Veterinary Teaching Hospital for surgical management.


A shoulder to cry on: Heart rate variability and empathetic behavioral responses to crying and laughing in dogs.
Dogs have a reputation for empathy toward their owners, which is also supported by some research (e.g., Carballo et al., 2020; Sanford, Burt, & Meyers-Manor, 2018). Many dog owners anecdotally report that dogs comfort them by making visual and/or physical contact when they cry or help them when they are sick. These behaviours provide a good way to assess the capacity for empathy and its physiological correlates in dogs. This study is a replication and extension of Custance and Mayer (2012). We examined whether using laughing as an alternative stimulus to humming produced similar responses to crying. Dogs were tested in their homes while a stranger and owner pretended to cry, laugh, and while the owner and stranger were talking. During each counterbalanced condition, the dog was observed for person-oriented behaviours and simultaneously had their heart rate variability measured. Like Custance and Mayer, dogs showed more behaviours directed toward the person crying, whether the owner or the stranger, than during baseline or laughing conditions. We did not find an effect of laughing on person-oriented behaviours, suggesting that dogs respond to the crying uniquely and not as a novel stimulus. In the condition when the stranger was crying, dogs that showed higher stress responses, as indicated by lower heart rate variability, were most likely to show person-oriented behaviours toward the stranger. This suggests that dogs that experience more distress, through emotional contagion, are more likely to show person-oriented behaviours toward the distressed stranger, indicating a possible mechanism for empathy-like behaviours. (PsycInfo Database Record (c) 2020 APA, all rights reserved).


The Bohr effect of haemoglobin in vertebrates: an example of molecular adaptation to different physiological requirements.
The Bohr effect, i.e. the pH dependence of the oxygen affinity of haemoglobins (Hbs) from a variety of vertebrates, and its modulation by temperature and other heterotropic effectors has been reviewed. Haemoglobins from vertebrates were not reviewed following the usual classification (i.e. mammals, birds, etc.); instead we have selected several key examples of animals, which are confronted with a similar environmental situation therefore displaying a similar life style. Hence, the paper starts from a description of the general concepts at the basis of the Bohr effect as exemplified by human HbA and goes towards the analysis of the modulation mechanisms which have been observed in different animals in response to the needs induced by: (i) life in cold environments; (ii) diving behaviour; (iii) flight; and (iv) aquatic life. The emerging picture indicates a complex organization of the information contained in the Hb molecule, the oxygen-binding properties of which depend both on the intrinsic characteristics of the protein and on its heterotropic interactions with ligands such as protons (Bohr effect), small anions like chloride and organic phosphates. In addition, each one of the functional effects induced by binding of a given effector appears to be under the strict control of temperature that enhances or decreases its relative weight with respect to all the others. It is just by this sophisticated network of interactions that the Hb molecule is able to satisfy the physiological requirements of a multitude of organisms without changing dramatically its quaternary structure.


Optimizing training opportunities for otolaryngology senior house officers: an audit.
Implementation of the European Working Time Directive and the Modernising Medical Careers initiative will mean junior surgeons must be trained in fewer hours over a shorter period. For this reason, junior surgeon training opportunities must be optimized. We undertook a departmental audit to identify where opportunities to train senior house officers (SHOs) in theatre were being lost, so that appropriate timetable changes could be made in order to optimize exposure to suitable surgical cases. During the first audit cycle, the SHOs followed their existing timetable and theatre attendance was monitored prospectively over a two-week period. Only 30 per cent of theatre sessions were attended and case participation was only 27 per cent. Simple timetable changes were made to maximize SHO theatre attendance, and a second prospective two-week audit was undertaken. The new rota yielded 46 per cent theatre attendance and 48 per cent case participation.


Reconstruction of orbital floor fractures using autologous nasal septal bone graft.
We describe herein a new technique for reconstruction of the orbital floor, using autologous nasal septal bone and report the surgical results achieved in maxillofacial trauma patients.Prior to its clinical surgical application, a cadaver practice was carried out on 5 formalin-fixed adult human cadavers to establish the feasibility and efficacy of the method. Fifteen patients with orbital floor fractures, operated between 2005 and 2008, using this technique, were included in the current study.Cadaveric practice revealed that an adequate and appropriate size of septal bone graft can be harvested for reconstruction of the orbital floor. All patients except one had satisfactory clinical and radiologic late results. One patient experienced persistent enophthalmos, possibly due to delayed repair and associated displaced zygomatic bone fracture.Autologous nasal septal bone as an orbital floor bone graft has many advantages, including low donor site morbidity, adequacy and appropriateness of size, and similarity of its bicortical morphology and histologic nature compared with the orbital floor bone. Our clinical results strongly support that this technique can become a satisfactory alternative to existing reconstruction methods.


Microarray Analysis of 8p23.1 Deletion in New Patients with Atypical Phenotypical Traits.
We describe two patients carrying deletions of chromosome 8p23.1 with a commonly critical region identified by means of oligonucleotide array comparative genomic hybridization (array CGH). They didn't present congenital heart defects or behavioral problems. Only one patient presented with intellectual disability and carrying deletion of TNKS gene. We presumed the inclusion of TNKS gene in the mental impairment.


Crush and smear technique for rapid detection and semiquantitation of amyloid deposition.
A method using Congo red to rapidly identify and semiquantitate amyloid deposits in tissues for experimental research and clinical medicine is described. Examination by polarization microscopy revealed amyloid deposits as bright green birefringent clumps on a dark red background. On semiquantitative evaluation, good correlation was found between this technique and the conventional histological one, the present technique being more sensitive. The method described saves time and expense.


PET/CT Imaging of Activated Cancer-Associated Fibroblasts Predict Response to PD-1 Blockade in Gastric Cancer Patients.
Promising development in immune checkpoint blockade (ICB) therapy has shown remarkable results in the treatment of gastric cancer (GC). However, the objective response rate in GC remains unsatisfactory. Noninvasive imaging to predict responses to ICB therapy via tumor microenvironment (TME) assessment is needed. Accordingly, this study aimed to evaluate the role of 68Ga-FAPI-04 PET/CT in the assessment of the immunosuppressive TME in GC and to cross-correlate imaging findings with responses to ICB therapy. The correlation between fibroblast-activation-protein (FAP) expression and immunosuppressive cell infiltration was analyzed using The Cancer Genome Atlas (TCGA), Gene Expression Omnibus (GEO) database, and GC tissue microarrays. To characterize the TME, TMEscores were calculated based on RNA-seq data from four GC patients. A total of 21 patients with GC underwent 68Ga-FAPI-04 PET/CT before ICB treatment, and two of them were imaged after ICB therapy. FAP expression was found to be closely correlated with poor prognosis and infiltration of immunosuppressive cells, including myeloid-derived suppressor cells (MDSCs), exhausted T cells, and regulatory T cells (Tregs) in GC. We also found a strong relationship (R 2 = 0.9678, p = 0.0162) between 68Ga-FAPI-04 uptake and TMEscore. Further analyses indicated that high 68Ga-FAPI-04 uptake was correlated with reduced therapeutic benefits from ICB therapy. 68Ga-FAPI-04 PET/CT may be used to noninvasively image the cancer-associated fibroblasts immunosuppressive TME in vivo and also potentially serve as a predictive biomarker of survival and antitumor immune response among patients who received ICB therapies.


Corrected cerebral blood flow and reduced cerebral inflammation in berk sickle mice with higher fetal hemoglobin.
Fetal hemoglobin (HbF) is known to lessen the severity of sickle cell disease (SCD), through reductions in peripheral vaso-occlusive disease and reduced risk for cerebrovascular events. However, the influence of HbF on oxygen delivery to high metabolism tissues like the brain, or its influence on cerebral perfusion, metabolism, inflammation or function have not been widely studied. We employed a Berkley mouse model (BERK) of SCD with gamma transgenes q3 expressing exclusively human α- and βS-globins with varying levels of γ globin expression to investigate the effect of HbF expression on the brain using magnetic resonance imaging (MRI), MRI diffusion tensor imaging (DTI) and spectroscopy (MRS) and hematological parameters. Hematological parameters improved with increasing γ level expression, as did markers for brain metabolism, perfusion and inflammation. Brain microstructure assessed by DTI fractional anisotropy improved, while myo-inositol levels increased, suggesting improved microstructural integrity and reduced cell loss. Our results suggest that increasing γ levels not only improves sickle peripheral disease, but also improves brain perfusion and oxygen delivery while reducing brain inflammation while protecting brain microstructural integrity.


[Sexual specificity of change of pancreatic alpha-amylase activity in rats in cadmium intoxication].
It was shown that the activity of alpha-amylase in the pancreas tissue, intestine content and blood, as well as kinetic characteristics in different substratum concentration changed in males and females after unitary oral administration of chloride cadmium in the dose 5 mg/kg. It was revealed that, after a single administration of chloride cadmium, growth of the pancreas weight, contents of protein and activity of alpha-amylase in the pancreatic tissue increased with parallel reduction of activity of enzyme in the small intestine. In acute cadmium intoxication, functional reorganizations in the pancreas are identical to initial stages of carcinogenic shifts. Such shifts are more expressed in females in comparison with males, especially after the cadmium intoxication.


A novel strategy for the detection of boldenone undecylenate misuse in cattle using ultra-high performance liquid chromatography coupled to high resolution orbitrap mass spectrometry: From non-targeted to targeted.
In this work multivariate strategies were employed in order to highlight new potential biomarkers of interest to detect the exogenous treatment of steers intramuscularly treated with boldenone undecylenate. Serum samples collected from treated (n = 4) and control (n = 8) crossbred animals of varying ages and weights were extracted using a simple sample preparation procedure based on salt assisted liquid-liquid extraction. Data acquisition was performed using liquid chromatography and Q-Exactive™ Orbitrap mass spectrometry. Data processing and treatment were performed using two non-targeted workflows: (1) Compound Discoverer software and (2) XCMS package on the open-source R software combined with MetaboAnalyst. Three potential biomarkers were highlighted taking into account the chromatographic shapes, the feature location on the generated s-plots, the fold change, the adjusted p values, the coefficient of variation in the QC samples and the area under the ROC curves. Predicted formulas based on mass accuracy, structural composition and spectra similarity were proposed. A robust statistical model to predict the boldenone treatment was further developed based on the weighted abundances of the selected biomarkers. The requirements for screening methods were successfully fulfilled, together with a wider detection window in comparison with the monitoring of the deconjugated metabolite boldenone, although biomarker identification studies are still ongoing.


Telomerase activation as a possible mechanism of action for psychopharmacological interventions.
Originally studied in relation to aging and cancer research, telomerase is now also investigated in relation to psychiatric disorders and treatments. Based on emerging clinical and preclinical data, we hypothesise that telomerase activation could represent a novel element mediating the mechanism of action of certain psychopharmacological interventions (e.g. antidepressants, lithium and antipsychotics). The modulation of intracellular Wnt/β-catenin or PI3K/Akt signalling pathways, the interaction with BDNF and 5-HT, and the antioxidant properties could represent possible mechanisms by which the different types of psychiatric medications could modulate telomerase activity. The potential of telomerase in promoting cellular survival and/or function in the brain and in the periphery could, in turn, represent a neurobiological substrate through which the enzyme can mediate the therapeutic effect of such interventions.


Vitamin D and breast cancer: interpreting current evidence.
Preclinical investigations and selected clinical observational studies support an association between higher vitamin D intake and 25-hydroxyvitamin D levels with lower breast cancer risk. However, the recently updated report from the Institute of Medicine concluded that, for cancer and vitamin D, the evidence was 'inconsistent and insufficient to inform nutritional requirements'. Against this background, reports examining vitamin D intake, 25-hydroxyvitamin D levels and breast cancer incidence and outcome were reviewed. Current evidence supports the pursuit of several research questions but not routine 25-hydroxyvitamin D monitoring and vitamin D supplementation to reduce breast cancer incidence or improve breast cancer outcome.


In vivo relaxometry of three brain tumors in the rat: effect of Mn-TPPS, a tumor-selective contrast agent.
T1 and T2 were determined simultaneously in vivo at 4.7 T in implanted rat brain tumors. Three different tumor cell lines were implanted in the right caudate nucleus: the F98 glioma, the E367 neuroblastoma, and the RN6 schwannoma. Their T1 and T2 values (mean +/- standard deviation [msec]), respectively, were 1,312 +/- 107 and 89 +/- 3 (glioma), 1,284 +/- 86 and 87 +/- 7 (neuroblastoma), and 1,338 +/- 85 and 86 +/- 9 (schwannoma). The T1 values (msec) of normal brain and muscle were 1,090 +/- 59 and 1,139 +/- 77, respectively, and the T2 values (msec) were 76 +/- 3 and 36 +/- 2, respectively. After injection of the contrast agent manganese (III) tetraphenylporphine sulfonate (TPPS) the T1 of all three tumors decreased by 30% and the T2 by 10%, whereas no such change in relaxivity was noted in normal brain. As a result, strong contrast enhancement of the three tumor types was seen on T1-weighted images. The tumor was clearly delineated and correlated with findings at histologic examination. This tumor enhancement was followed up for 4 days with quantitative relaxation time measurements, and the strong, selective reduction in T1 for all three tumor types after Mn-TPPS injection was preserved over the entire observation period.


A putative Vibrio cholerae two-component system controls a conserved periplasmic protein in response to the antimicrobial peptide polymyxin B.
The epidemic pathogen Vibrio cholerae senses and responds to different external stresses it encounters in the aquatic environment and in the human host. One stress that V. cholerae encounters in the host is exposure to antimicrobial peptides on mucosal surfaces. We used massively parallel cDNA sequencing (RNA-Seq) to quantitatively identify the transcriptome of V. cholerae grown in the presence and absence of sub-lethal concentrations of the antimicrobial peptide polymyxin B. We evaluated the transcriptome of both wild type V. cholerae and a mutant carrying a deletion of vc1639, a putative sensor kinase of an uncharacterized two-component system, under these conditions. In addition to many previously uncharacterized pathways responding with elevated transcript levels to polymyxin B exposure, we confirmed the predicted elevated transcript levels of a previously described LPS modification system in response to polymyxin B exposure. Additionally, we identified the V. cholerae homologue of visP (ygiW) as a regulatory target of VC1639. VisP is a conserved periplasmic protein implicated in lipid A modification in Salmonellae. This study provides the first systematic analysis of the transcriptional response of Vibrio cholerae to polymyxin B, raising important questions for further study regarding mechanisms used by V. cholerae to sense and respond to envelope stress.


Gas chromatography/combustion/isotope ratio mass spectrometric comparison of N-acetyl- and N-pivaloyl amino acid esters to measure 15N isotopic abundances in physiological samples: a pilot study on amino acid synthesis in the upper gastro-intestinal tract of minipigs.
There is evidence that in animals and also in humans, non-specific nitrogen is used for de novo synthesis of indispensable amino acids by the microflora of the gastro-intestinal tract. Gas chromatography/combustion/isotope ratio mass spectrometry was applied to investigate whether lysine of intestinal origin is available for body protein synthesis. Two minipigs with an end-to-end ileorectal anastomosis received orally equimolar amounts of 15N as ammonium chloride or urea twice a day for 10 days. Samples of blood and ileal digesta were collected before and throughout the 10 days; tissue samples were taken at termination of the experiment. The N-acetyl-n-propyl (NAP)- and N-pivaloyl-i-propyl (NPP)-amino acid esters were evaluated for the determination of the 15N content of lysine and 16 other amino acids ranging from natural abundance to an enrichment of 0.6 APE 15N in a complex mixture of proteinogenic amino acids and several matrices. At natural abundances for all amino acids analysed, NAP and NPP derivatives gave mean precisions of 0.5 and 0.3/1000 delta 15N, respectively. The mean precision for NPP derivatives at enrichments between 0.42 and 1.10 AP 15N ranged between 1.0-15.0/1000 delta 15N. 15N from ammonium chloride was incorporated into lysine and in all other amino acids of serum albumin to a 2.5 times higher degree than from urea. Somewhat lower, but significant, lysine enrichments were detected in liver, duodenum and jejunum. After 10 days of ingestion of 15N-labeled urea a significant 15N enrichment in lysine of serum albumin could not be detected, although lysine in the ileal digesta was significantly labeled by day 5. This is the first report providing evidence that the microflora in the upper gastrointestinal tract of pigs is capable of synthesizing lysine de novo and that this lysine is available for body protein synthesis.


Cross-national Differences in Intergenerational Family Relations: The Influence of Public Policy Arrangements.
Focusing mostly on Europe, this overview reveals how the research on cross-national differences in intergenerational family relations has moved from basic descriptions to a focus on understanding how support exchanges are shaped by macro-level processes. A key issue concerns generational interdependence, the extent to which public policy arrangements impose reliance on older and younger family members or enable individual autonomy. Real theoretical progress is visible in three areas of research. The first pertains to analyses at the micro level of how family members actually respond to the incentives that different macro contexts provide. The generosity or restrictedness of public provisions variably releases or necessitates normative obligations in interdependent family relationships. The second area of progress involves analyses of the implications of specific policies rather than policy packages for gender and socioeconomic inequality. The third area of progress is a more nuanced view on the familialism-individualism divide. These three areas provide inspiring examples for future investigations.


Chronic oedema: impact and management.
Chronic oedema of all causes is thought to be more prevalent than studies have shown. The impact of this chronic condition encompasses many physical, psychological and social domains, many of which are not acknowledged nor addressed. Successful management relies on three factors. The first is early detection and commencement of treatment to prevent complications occurring. The second is an holistic multidimensional approach to address all the associated physical, psychological and social sequellae. Finally, collaboration between specialist and generalist health professionals is essential to ensure the best treatment is delivered in the best healthcare location. To achieve these there needs to be a network of suitably qualified practitioners who are upskilled by robust awareness and education systems, all of which should be supported by stable financial and human resources.


Alkylated Aromatic Thioethers with Aggregation-Induced Emission Properties-Assembly and Photophysics.
In this contribution, we present the synthesis and self-assembly of alkylated thioethers with interesting photophysical properties. To this end, the emission, absorption and excitation spectra in organic solvents and as aggregates in water were measured as well as the corresponding photoluminescence quantum yields and lifetimes. The aggregates in aqueous media were visualized and measured using transmission electron microscopy. Besides that, crystal structures of selected compounds allowed a detailed discussion of the structure-property relationship. Furthermore, the mesomorphic behavior was investigated using polarized optical microscopy (POM) as well as differential scanning calorimetry (DSC).


Comparison of the NAD Kinase and Myosin Light Chain Kinase Activator Properties of Vertebrate, Higher Plant, and Algal Calmodulins.
In the preceding paper (Lukas, Iverson, Schleicher, Watterson 1984 Plant Physiol 75: 788-795), we reported that the amino acid sequence of spinach calmodulin has at least 13 amino acid sequence differences from vertebrate calmodulin. In the present study, we investigated the effect of these amino acid sequence substitutions on the enzyme activator properties of vertebrate and plant calmodulins. Calmodulins from spinach and the green alga Chlamydomonas reinhardtii activate chicken gizzard myosin light chain kinase in a manner similar but not identical to chicken calmodulin. In contrast, these calmodulins have very different NAD kinase activator properties. The concentration required for half-maximal activation of pea seedling NAD kinase by spinach calmodulin (3-4 nanomolar) is lower than the corresponding concentrations of chicken (20 nanomolar) and Chlamydomonas (40 nanomolar) calmodulins. However, the maximum level of activation obtained with Chlamydomonas calmodulin is 4- to 6-fold higher than spinach or chicken calmodulin. These data indicate that the limited structural heterogeneity among calmodulins have differential effects on their biochemical activities.


Structural studies of the carbon monoxide complex of [NiFe]hydrogenase from Desulfovibrio vulgaris Miyazaki F: suggestion for the initial activation site for dihydrogen.
The carbon monoxide complex of [NiFe]hydrogenase from Desulfovibrio vulgaris Miyazaki F has been characterized by X-ray crystallography and absorption and resonance Raman spectroscopy. Nine crystal structures of the [NiFe]hydrogenase in the CO-bound and CO-liberated forms were determined at 1.2-1.4 A resolution. The exogenously added CO was assigned to be bound to the Ni atom at the Ni-Fe active site. The CO was not replaced with H(2) in the dark at 100 K, but was liberated by illumination with a strong white light. The Ni-C distances and Ni-C-O angles were about 1.77 A and 160 degrees, respectively, except for one case (1.72 A and 135 degrees ), in which an additional electron density peak between the CO and Sgamma(Cys546) was recognized. Distinct changes were observed in the electron density distribution of the Ni and Sgamma(Cys546) atoms between the CO-bound and CO-liberated structures for all the crystals tested. The novel structural features found near the Ni and Sgamma(Cys546) atoms suggest that these two atoms at the Ni-Fe active site play a role during the initial H(2)-binding process. Anaerobic addition of CO to dithionite-reduced [NiFe]hydrogenase led to a new absorption band at about 470 nm ( approximately 3000 M(-1)cm(-1)). Resonance Raman spectra (excitation at 476.5 nm) of the CO complex revealed CO-isotope-sensitive bands at 375/393 and 430 cm(-1) (368 and 413 cm(-1) for (13)C(18)O). The frequencies and relative intensities of the CO-related Raman bands indicated that the exogenous CO is bound to the Ni atom with a bent Ni-C-O structure in solution, in agreement with the refined structure determined by X-ray crystallography.


A natural language processing and deep learning approach to identify child abuse from pediatric electronic medical records.
Child physical abuse is a leading cause of traumatic injury and death in children. In 2017, child abuse was responsible for 1688 fatalities in the United States, of 3.5 million children referred to Child Protection Services and 674,000 substantiated victims. While large referral hospitals maintain teams trained in Child Abuse Pediatrics, smaller community hospitals often do not have such dedicated resources to evaluate patients for potential abuse. Moreover, identification of abuse has a low margin of error, as false positive identifications lead to unwarranted separations, while false negatives allow dangerous situations to continue. This context makes the consistent detection of and response to abuse difficult, particularly given subtle signs in young, non-verbal patients. Here, we describe the development of artificial intelligence algorithms that use unstructured free-text in the electronic medical record-including notes from physicians, nurses, and social workers-to identify children who are suspected victims of physical abuse. Importantly, only the notes from time of first encounter (e.g.: birth, routine visit, sickness) to the last record before child protection team involvement were used. This allowed us to develop an algorithm using only information available prior to referral to the specialized child protection team. The study was performed in a multi-center referral pediatric hospital on patients screened for abuse within five different locations between 2015 and 2019. Of 1123 patients, 867 records were available after data cleaning and processing, and 55% were abuse-positive as determined by a multi-disciplinary team of clinical professionals. These electronic medical records were encoded with three natural language processing (NLP) algorithms-Bag of Words (BOW), Word Embeddings (WE), and Rules-Based (RB)-and used to train multiple neural network architectures. The BOW and WE encodings utilize the full free-text, while RB selects crucial phrases as identified by physicians. The best architecture was selected by average classification accuracy for the best performing model from each train-test split of a cross-validation experiment. Natural language processing coupled with neural networks detected cases of likely child abuse using only information available to clinicians prior to child protection team referral with average accuracy of 0.90±0.02 and average area under the receiver operator characteristic curve (ROC-AUC) 0.93±0.02 for the best performing Bag of Words models. The best performing rules-based models achieved average accuracy of 0.77±0.04 and average ROC-AUC 0.81±0.05, while a Word Embeddings strategy was severely limited by lack of representative embeddings. Importantly, the best performing model had a false positive rate of 8%, as compared to rates of 20% or higher in previously reported studies. This artificial intelligence approach can help screen patients for whom an abuse concern exists and streamline the identification of patients who may benefit from referral to a child protection team. Furthermore, this approach could be applied to develop computer-aided-diagnosis platforms for the challenging and often intractable problem of reliably identifying pediatric patients suffering from physical abuse.


Captopril inhibits pp60(c-src) tyrosine phosphorylation in cultured human mesangial cells.
The present experiments were devoted to analyzing the mechanisms involved in the captopril-dependent inhibition of human mesangial cell proliferation. Studies were performed in cultured human mesangial cells incubated with captopril, an angiotensin II-converting enzyme inhibitor with antioxidant properties, lisinopril, a non-antioxidant angiotensin II-converting enzyme inhibitor, and tocopherol, a pure antioxidant. Both angiotensin II-converting enzyme inhibitors significantly inhibited fetal calf serum-induced [3H]thymidine uptake by human mesangial cells, in a dose- and time-dependent manner, an effect which was not observed with tocopherol. The antiproliferative effect of captopril and its ability to block tyrosine phosphorylation of human mesangial cells proteins were significantly greater than those of lisinopril. Moreover, captopril significantly prevented the fetal calf serum-induced tyrosine phosphorylation of pp60(c-src). The present results suggest that the antiproliferative ability of captopril does not completely depend on its angiotensin II-converting enzyme inhibitor properties, pointing to a possible interaction of the drug with the intracellular mechanisms responsible for the transmission of the proliferative signals.


[Surgical treatment of intramedullary tumors: preliminary results].
Since 1995 in the Department of Neurosurgery, Mining Hospital in Sosnowiec a surgical technique has been used of total or gross total removal of tumour. It includes the use of CO2 laser for myelotomy and tumour vaporisation as well as ultrasonic aspirator, microsurgical techniques and intraoperative monitoring of SSEP. Ten patients 11-61 years of age were operated on. Four tumours were located in the cervical part of the spinal cord, three in the cervico-thoracic area while the remaining three in the thoracic part. The condition of four patients before the operation was assessed as I-II degree on the McCormick scale, of one patient as--III degree--and of the other five patients as IV-V degree. Only in two cases the removed tumour was ependymoma; in six cases these were benign astrocytomas and in two other--anaplastic tumours. After the treatment, the clinical condition of almost all patients improved. Pain and trouble some paraesthesias subsided. Five patients are completely self-dependent and three other walk with the aid of handrails. The treatment proved unsuccessful in one patient with an anaplastic tumour in whom paralysis of lower limbs occurred before the operation. Six months after the operation the patient died of tumour metastasis. The observation period ranged from 6 months to 3 years. On the basis of our own observations it is suggested that total safe removal of even infiltrating intramedullary tumours is possible when proper surgical techniques are used. Best results are obtained in patients in good clinical condition irrespective of tumour extent.


Far-red switching DNA probes for live cell nanoscopy.
Herein we present DNA probes composed of Hoechst 33258 and spontaneously blinking far-red hydroxymethyl silicon-rhodamine (HMSiR). The best performing probe, 5-HMSiR-Hoechst, contains the 5'-regioisomer, shows ∼400-fold fluorescence increase upon DNA binding and is compatible with wash-free single molecule localization and 3D stimulated emission depletion microscopy of chromatin nanostructures in living cells.


Responsiveness to D-glucose in neurosecretory cells of crustaceans.
1. An electrophysiological study of the D-glucose sensitivity of X-organ (XO) neurosecretory cell bodies in crayfish was carried out with the use of microelectrodes, perforated, and cell-attached patch-clamp techniques. 2. Glucose depolarizes the membrane potential of XO cells in a concentration-dependent manner. 3. Depolarization produced by glucose initiates a change in the pattern of electrical activity. Silent cells began to discharge action potentials. When bursting cells are depolarized by glucose, their action potentials are no longer grouped in bursts or disappear entirely. 4. Although the membrane potential returns to its initial value after removing glucose from the bath, discharge patterns of the cells may remain different. This suggests that besides the depolarizing effect, once the cells have been exposed to glucose, the sugar switches on a process that is maintained for a long time. 5. Glucose produced a reduction of membrane steady-state conductance, and a shift of reversal potential of membrane currents to a more positive value. 6. Depolarization induced by D-glucose appears to be related with a closure of potassium channels. 7. Glucose effect was thought to be generated by a product of metabolism that would act as intracellular mediator.


Immunologic and Allergic Properties of Cows' Milk Proteins in Humans.
Cows' milk is an important component of the diet especially during infancy. Yet, cows' milk can elicit allergic and other sensitivity reactions in some individuals. Cows' milk allergy (CMA) results from an abnormal immunologic reaction to cows' milk proteins. IgE responses are definitely involved in CMA. Immune complexes and tissue lymphocytes may also play a role in some forms of CMA, but further evidence is needed to firmly establish this possibility. The presence of circulating antibodies to cows' milk proteins of the IgG, IgA and IgM classes is not clinically significant. Such antibodies are found in both normal and allergic individuals. (β-Lactoglobulin and casein are the most common cows' milk allergens, although other cows' milk proteins may play important roles in some cases. Partial digestion of cows' milk proteins may enhance their allergenicity, whereas complete hydrolysis abolishes their allergenicity. Heating can also alter the allergenicity of the cows' milk proteins, but rather severe heating is required. More research will be necessary to develop hypoallergenic processing methods for cows' milk.