Complex interventions to improve physical function and maintain independent living in elderly people: a systematic review and meta-analysis.
In old age, reduction in physical function leads to loss of independence, the need for hospital and long-term nursing-home care, and premature death. We did a systematic review to assess the effectiveness of community-based complex interventions in preservation of physical function and independence in elderly people. We searched systematically for randomised controlled trials assessing community-based multifactorial interventions in elderly people (mean age at least 65 years) living at home with at least 6 months of follow-up. Outcomes studied were living at home, death, nursing-home and hospital admissions, falls, and physical function. We did a meta-analysis of the extracted data. We identified 89 trials including 97 984 people. Interventions reduced the risk of not living at home (relative risk [RR] 0.95, 95% CI 0.93-0.97). Interventions reduced nursing-home admissions (0.87, 0.83-0.90), but not death (1.00, 0.97-1.02). Risk of hospital admissions (0.94, 0.91-0.97) and falls (0.90, 0.86-0.95) were reduced, and physical function (standardised mean difference -0.08, -0.11 to -0.06) was better in the intervention groups than in other groups. Benefit for any specific type or intensity of intervention was not noted. In populations with increased death rates, interventions were associated with reduced nursing-home admission. Benefit in trials was particularly evident in studies started before 1993. Complex interventions can help elderly people to live safely and independently, and could be tailored to meet individuals' needs and preferences.


Serological biomarkers detect active joint destruction and inflammation in patients with haemophilic arthropathy.
Progressive arthropathy caused by recurrent joint bleeds is a severe complication in haemophilia. We investigated whether biomarkers of cartilage and bone degradation, and inflammation were altered in haemophilia patients and whether these biomarkers could identify haemophilia patients with arthropathy. Serum from 35 haemophilia patients with varying degrees of arthropathy and 43 age- and gender-matched control subjects were analysed. Biomarkers of cartilage degradation (C2M, COMP, CTX-II, ADAMTS5), cartilage formation (PRO-C2), bone formation (PINP), bone resorption (CTX-I) and inflammation (hsCRP, CRPM) were measured by ELISA. Arthropathy was assessed by radiological evaluation (Pettersson score) and physical examination (Gilbert score). In patients with haemophilia, cartilage degradation, measured by C2M, CTX-II and COMP, was increased by 25% (P < 0.05) compared with control subjects. Levels of the cartilage degradation enzyme, ADAMTS5, were 10% lower in haemophilia patients (P < 0.05). Bone formation (PINP) was reduced by 25% (P < 0.05) in haemophilia patients, whereas bone resorption (CTX-I) was increased by 30% (P < 0.001). Acute inflammation (hsCRP) was increased by 50% (P < 0.01), whereas chronic inflammation (CRPM) was decreased by 25% (P < 0.0001). The hsCRP/CRPM ratio was 60% higher (P < 0.001) in haemophilia patients relative to control subjects. A biomarker panel combining C2M, CRPM, and ADAMTS5 could distinguish haemophilia patients from control subjects with 85.3% accuracy (P < 0.0001). We found no strong correlation between biomarkers and radiological and physical examination of the joint. Biomarkers detect increased cartilage and bone degradation, and altered inflammatory activity in haemophilia patients with arthropathy. These biomarkers could potentially be used to identify patients with progressing joint disease.


The effect of disodium ethane-1-hydroxy-1,1-diphosphonate on the metabolism of calcitriol in chicks.
Decreased intestinal absorption of Ca2+ occurs in response to treatment with disodium ethane-1-hydroxy-1,1-diphosphonate (EHDP). The effect is due to decreased 1-hydroxylation of calcidiol (25-hydroxycholecalciferol) in the kidney. In an attempt to establish whether impairment of vitamin D metabolism at steps beyond kidney hydroxylation occurs due to treatment with EHDP, chicks were depleted of vitamin D and were treated with calcitriol (1,25-dihydroxycholecalciferol) as their sole source of the vitamin. The chicks were then divided into two groups, one being treated with EHDP while the second group served as control. Intestinal absorption of Ca2+ in the EHDP-treated group was found to be impaired, along with decreases in concentrations of calbindin D28K (the 28,000-Mr vitamin D-dependent Ca2+-binding protein). When the chicks were dosed with [3H]calcitriol, significantly lower concentrations of the sterol were detected in the duodena of EHDP-treated birds. Measurement of levels of receptors for calcitriol in duodena showed no difference between groups, but levels of calcitriol in sera were considerably lower in the EHDP-treated group along with the elevated biliary and urinary excretion of glucuronidated conjugates. It is therefore concluded that treatment with EHDP results in increased catabolism of calcitriol in addition to the known suppression of the renal production of the hormone.


Theoretical determination of spin Hamiltonians with isotropic and anisotropic magnetic interactions in transition metal and lanthanide complexes.
The determination of anisotropic magnetic parameters is a task of both experimental and theoretical interest. The added value of theoretical calculations can be crucial for analyzing experimental data by (i) allowing assessment of the validity of the phenomenological spin Hamiltonians, (ii) allowing discussion of the values of parameters extracted from experiments, and (iii) proposing rationalizations and magneto-structural correlations to better understand the relations between geometry, electronic structure, and properties. In this review, we discuss the model Hamiltonians that are used to describe magnetic properties, the computational approaches that can be used to compute magnetic parameters, and review their applications to transition metal and (to a lesser extent) lanthanide based complexes. Perspectives concerning current methodological challenges will then be presented, and finally the need for further joint experimental/theoretical efforts will be underlined.


Influence of Season and Diet on Fiber Digestion and Bacterial Community Structure in the Rumen of Muskoxen (Ovibos moschatus).
We studied the relationship between fiber digestion and the composition of the bacterial community in the rumen of muskoxen at the start and the end of the annual window of plant growth from spring to fall. Eight ruminally cannulated castrated males were fed brome hay or triticale straw (69.6% vs. 84.6% neutral detergent fiber, respectively) that were similar in fiber content to the sedges consumed by wild muskoxen (64.5 to 71.7% neutral detergent fiber). Muskoxen digested fiber from both forages faster and to a greater extent when straw rather than hay was consumed. Fiber digestion was therefore inducible by diet 4 in each season. We used 16S rRNA sequences from ruminal contents to study how season and diet affected the bacterial community and how the latter related to fiber digestion. We found that Bacteroidetes and Firmicutes accounted for 90% of the sequences at the level of Phylum, which is typical for the mammal gut microbiome. Using partial least square regressions, it was found that between 48% and 72% of the variation in fiber digestion was associated with 36⁻43 genera of bacteria. The main fibrolytic bacteria typical of domestic ruminants were generally not among the most important bacteria associated with fiber digestion in muskoxen. This reveals that muskoxen rely upon on a large suite of bacterial genera that are largely distinct from those used by other ruminants to digest the cell walls of plants that vary widely in both abundance and nutritional quality through the year.


The role of sludge conditioning and dewatering in the fate of nonylphenol in sludge-amended soils.
One of the main concerns associated with the recycling of biosolids to arable land is their contamination by organic pollutants, like endocrine disruptors. Conditioning and dewatering are usually the last steps of the sewage sludge treatment, before its further utilization. The choice of the specific conditioning/dewatering method may have an effect, not only on the amount of residues in the biosolids, but also on the fate of these compounds in amended soils. Anaerobically digested wastewater sludge was conditioned at lab-scale by means of physical and chemical methods and subsequently dewatered by centrifugation. The produced biosolids plus non-conditioned and non-dewatered sludges were amended separately to soil and spiked with 14C radiolabelled single isomer of nonylphenol. The persistence and leaching potential of nonylphenol after an incubation period of three months were correlated to the sludge treatment method. In comparison to non-conditioned sludge, 54% and 72% higher amount of pollutant residues were extractable when freeze-thawed and limed sludge, respectively, were used. Conditioning of sludge with cationic polymer decreased the leaching potential of nonylphenol in sludge-amended soils, while liming increased it. Fractions of the model compound recovered as extractable and bound residues were analyzed in order to interpret nonylphenol fate.


[Significance of ultrasonics in the placement of a central venous catheter].
An ultrasound investigation was undertaken of the neck region of 42 patients with normal neck anatomy in order to determine whether the results of ultrasound-gained topographical data provided pointers to the choice of entry site to the internal jugular vein (IJV). In addition, the IJV was punctured under ultrasound control in 23 patients in an intensive care unit in whom there was a problem of increased bleeding tendency, anatomical difficulty or previously failed "blind" puncture. In all of them a central venous catheter was placed without complication by the Seldinger technique via the primary chosen point for puncture. An approach through the sternocleidomastoid muscle, between the cricoid level and the "central" place of puncture between the two bellies of the sternocleidomastoid muscle proved to be the most satisfactory compromise between easy application of the ultrasound head, large vein diameter and reduction of any risk of mistakenly puncturing artery or pleura. This approach has to be varied according to the ultrasound findings. It is concluded from this experience that ultrasound is suitable for the placement of central venous catheters. But since the equipment is bulky it cannot be used in an emergency.


Leptin stimulates ischemia-induced retinal neovascularization: possible role of vascular endothelial growth factor expressed in retinal endothelial cells.
Diabetic retinopathy is the leading cause of new blindness in adults in developed countries. Leptin, an adipocyte-derived hormone, stimulates endothelial proliferation and angiogenesis. This study was designed to elucidate the pathophysiologic role of leptin in the progression of retinal neovascularization. Using the retinopathy of prematurity model, a mouse model of ischemia-induced retinal neovascularization, we have demonstrated more pronounced retinal neovascularization in 17-day-old transgenic mice overexpressing leptin than in age-matched wild-type littermates. Ischemia-induced retinal neovascularization was markedly suppressed in 17-day-old leptin-deficient ob/ob mice. Western blot analysis revealed that a biologically active leptin receptor isoform is expressed in mouse retinal endothelial cells. Leptin receptor expression was also detected in primary cultures of porcine retinal endothelial cells, where it upregulated vascular endothelial growth factor (VEGF) mRNA expression. This effect was thought to be mediated at least partly through the activation of signal transducers and activators of transcription (STAT)3, because adenoviral transfection of the dominant-negative form of STAT3 abolished the leptin-induced upregulation of VEGF mRNA expression in retinal endothelial cells. This study provides evidence that leptin stimulates the ischemia-induced retinal neovasucularization possibly through the upregulation of endothelial VEGF, thereby suggesting that leptin antagonism may offer a novel therapeutic strategy to prevent or treat diabetic retinopathy.


Transmission fourier transform Raman spectroscopy of pharmaceutical tablet cores.
Transmission Fourier transform (FT) Raman spectroscopy of pharmaceutical tablet cores is demonstrated using traditional, unmodified commercial instrumentation. The benefits of improved precision over backscattering Raman spectroscopy due to increased sample volume are demonstrated. Self-absorption effects on analyte band ratios and sample probe volume are apparent, however. A survey of near-infrared (NIR) absorption spectra in the FT-Raman spectral range (approximately 0 to 3500 wavenumber shift from 1064 nm, or 1064 to 1700 nm) of molecules with a wide range of NIR-active functional groups shows that although absorption at the laser wavelength (1064 nm) is relatively small, some regions of the Raman spectrum coincide with NIR absorbances of 0.5 per cm or greater. Fortunately, the pharmaceutically important regions of the Raman shift spectrum from 0 to 600 cm(-1) and from 1400 to 1900 cm(-1) exhibit low self-absorption for most organic materials. A statistical analysis of transmission FT-Raman noise in spectra collected from different regions of a pharmaceutical tablet provides insight into both spectral distortion and reduced sampling volume caused by self-absorption.


[EEG frequency: amplitude characteristics at different levels of consciousness].
Relationship was studied between the level of consciousness and the level of the brain activation. State of transition from wakefulness to drowsiness was used as a model of gradual decrease in the level of consciousness. A change in the intensity and quality of appearing involuntarily mental processes served as an index of the level of consciousness as determined by subjects' self-reports. It was found that a certain EEG profile corresponded to each level of consciousness under study. The data obtained showed that the lowest level of activation corresponded to a state in which a subject develops inhibition of "internal speech", "failure of thoughts". In this case, the dynamics of the EEG high frequency rhythm changes was possibly related to the mechanism of transmission of cognitive information.


Long-term outcome of patients with acute ulcerative colitis after first course of intravenous corticosteroids.
Every fifth patient with ulcerative colitis (UC) experiences severe acute flare at some point in the course of the disease. Corticosteroids (Cs) remain the treatment of choice in acute flare. Data on the efficacy of first intravenous Cs in the long-term prognosis of UC are scarce and were investigated here. All episodes of patients with acute UC admitted to Tampere University Hospital and treated with intravenous Cs between January 2007 and January 2016 were identified from patient records and reviewed. The risks for colectomy and for continuous use of Cs were evaluated. Predictive factors were analysed. The study comprised 217 patients of whom 184 (85%) responded to intravenous Cs at index flare. Of the 33 non-responders, 31 (94%) were treated with intravenous cyclosporine A and 28 responded. Five (2.3%) patients needed emergency colectomy. Twenty-six (12%) patients underwent colectomy within 1 year of index flare. Overall colectomy rate was 56 (26%) during follow-up (median 7.5 years, range 0.1-10.5). Six months after index flare 66 (30%) patients were still on steroids. In this series 149 (69%) required further Cstherapy and 104 (48%) needed rehospitalization for new flare at some point during follow-up. Overall 155 patients were treated with thiopurines, of whom 72% within the first year after admission. A total of 36 patients had infliximab as a first-line biological treatment, nine needed second-line therapy with adalimumab or vedolizumab after infliximab failed. Although intravenous Cs were efficient in inducing clinical response in patients with severe acute UC, only one fifth maintained remission in the long term. Two-thirds of patients required further Cs and the overall colectomy rate remained at 26%. High relapse rate indicates the need for closer monitoring of these patients. Enhancement of maintenance therapy should be considered at early stage after acute flare.


Effect of cell culture medium components on color of formulated monoclonal antibody drug substance.
As the industry moves toward subcutaneous delivery as a preferred route of drug administration, high drug substance concentrations are becoming the norm for monoclonal antibodies. At such high concentrations, the drug substance may display a more intense color than at the historically lower concentrations. The effect of process conditions and/or changes on color is more readily observed in the higher color, high concentration formulations. Since color is a product quality attribute that needs to be controlled, it is useful to study the impact of process conditions and/or modifications on color. This manuscript summarizes cell culture experiments and reports on findings regarding the effect of various media components that contribute to drug substance color for a specific monoclonal antibody. In this work, lower drug substance color was achieved via optimization of the cell culture medium. Specifically, lowering the concentrations of B-vitamins in the cell culture medium has the effect of reducing color intensity by as much as 25%. In addition, decreasing concentration of iron was also directly correlated color intensity decrease of as much as 37%. It was also shown that the color of the drug substance directly correlates with increased acidic variants, especially when increased iron levels cause increased color. Potential mechanisms that could lead to antibody coloration are briefly discussed.


Combined functional and morphological imaging consisting of gated myocardial perfusion SPECT and 16-detector multislice spiral CT angiography in the noninvasive evaluation of coronary artery disease: first experiences.
Appropriate diagnosis and therapy of coronary artery disease (CAD) frequently require information about both the functional and morphological status of the coronary artery tree. We hypothesized that the combination of multislice spiral CT (MDCT) angiography and myocardial perfusion SPECT (MPI) provides accurate allocation of perfusion defects (PD) to their determining coronary lesion. Twenty patients (14 male, mean age 64+/-9.2 years) with known CAD were retrospectively studied. Gated MPI, CT angiography using a 16-detector CT scanner, and conventional coronary angiography (CCA) were performed in each patient. Reversible and fixed PD were subsequently allocated to their determining lesion separately by different observers for MDCT angiography and CCA. All patients showed significant CAD in CCA; six patients with one-, six with two-, six with three-, and two with four-vessel disease; three patients had bypass grafts; and five patients had prior myocardial infarction. Correct diagnosis of CAD was stated in 14 of 20 patients by MDCT angiography. Five reversible and five fixed PD were detected in 9 of 20 patients; one patient showed both reversible and fixed PD. Five of five reversible PD could be allocated to appropriate coronary artery stenoses in CCA. In MDCT angiography, five of five reversible PD were allocated to the same lesions; all lesions were rated as >/=50%. The preliminary results of the present study show high accuracy for multislice spiral CT angiography to allocate reversible perfusion defects in myocardial scintigraphy to their determining coronary artery lesions in a small patient collective with known coronary artery disease.


Klebsiella pneumoniae and Colistin Susceptibility Testing: Performance Evaluation for Broth Microdilution, Agar Dilution and Minimum Inhibitory Concentration Test Strips and Impact of the "Skipped Well" Phenomenon.
The emergence of multidrug resistant Gram-negative pathogens, particularly carbapenemase producers, has forced clinicians to use last line antibiotics, such as colistin. Since colistin susceptibility testing presents several challenges, this study aimed at evaluating the performance of two alternative susceptibility methods for Klebsiella pneumoniae, namely, agar dilution (AD) and MIC test strips (MTS). These approaches were compared with the reference method, broth microdilution (BMD), and provide a quantitative description for the "skipped well" (SW) phenomenon. Colistin susceptibility was evaluated by BMD and AD in parallel and triplicate, using 141 K. pneumoniae clinical isolates while MTS performance was evaluated only for a subset (n = 121). Minimum inhibitory concentration analysis revealed that a substantial part (n = 26/141; 18.4%) of the initial isolates was deemed undetermined by BMD due to the following: discordance between replicates (1.4%); presence of multiple SWs (7.8%); and the combination of both events (9.2%). Both AD and MTS revealed a high number of false-susceptible strains ("very major errors"), 37.5% and 68.8%, respectively. However, AD agreement indices were reasonably high (EA = 71.3% and CA = 94.8%). For MTS these indices were lower, in particular EA (EA = 41.7% and CA = 89.6), but the approach enabled the detection of distinct sub-populations for four isolates. In conclusion, this study provides the most comprehensive study on the performance of AD and MTS for colistin susceptibility testing in K. pneumoniae, highlighting its limitations, and stressing the importance of sample size and composition. Further, this study highlights the impact of the SW phenomenon associated with the BMD method for K. pneumoniae.


Brief Report: Zinc Supplementation and Inflammation in Treated HIV.
In this study, we explored the effect of zinc supplementation on markers of inflammation and monocyte activation in antiretroviral therapy-treated HIV infection. This is a phase I open-labeled randomized double-arm study, exploring the efficacy and safety of zinc supplementation on inflammation in ≥18-year-old people living with HIV in the US, on stable antiretroviral therapy and with zinc levels ≤75 µg/dL in the last 60 days. Patients were randomized 1:1 to zinc gluconate capsules at a dose of 45 mg (low-dose), or 90 mg (high-dose) elemental zinc daily for 16 weeks. We assessed inflammatory and gut integrity biomarkers at baseline and 16 weeks. Overall, a total of 52 participants were enrolled (25 participants in the low-dose arm and 27 participants in the high-dose arm). Median (Interquartile range) age was 49 (38, 60) years, 77% were men and 73% were African Americans. At baseline, median zinc levels were 73 (64, 86) µg/dL. Median circulating zinc levels increased to 91 µg/dL in the low-dose arm and to 100 µg/dL in the high-dose arm. Overall, 48%-60% of participants experienced a reduction in biomarkers levels. The margin of reduction ranged between 8% and 21%. This change was meaningful with large effect size (Cohen D ranging from 5 to 19). In this pilot study, we found that zinc supplementation is effective at increasing circulating zinc levels. In addition, our findings provide novel data suggesting that zinc can affect a biological signature in people living with HIV and modulate biomarkers associated with clinical comorbidities.


Elastic Instability of Cubic Blue Phase Nano Crystals in Curved Shells.
Many crystallization processes, including biomineralization and ice-freezing, occur in small and curved volumes, where surface curvature can strain the crystal, leading to unusual configurations and defect formation. The role of curvature on crystallization, however, remains poorly understood. Here, we study the crystallization of blue phase (BP) liquid crystals under curved confinement, which provides insights into the mechanism by which BPs reconfigure their three-dimensional lattice structure to adapt to curvature. BPs are a three-dimensional assembly of high-chirality liquid crystal molecules arranged into body-centered (BPI) or simple cubic (BPII) symmetries. BPs with submicrometer cubic-crystalline lattices exhibit tunable Bragg reflection and submillisecond response time to external stimuli such as an electric field, making them attractive for advanced photonic materials. In this work, we have systematically studied BPs confined in spherical shells with well-defined curvature and boundary conditions. The optical behavior of shells has also been examined at room temperature, where the cholesteric structure forms. In the cholesteric phase, perpendicular anchoring generates focal conic domains on the shell's surface, which transition into stripe patterns as the degree of curvature increases. Our results demonstrate that both higher degrees of curvature and strong spatial confinement destabilize BPI and reconfigure that phase to adopt the structure and optical features of BPII. We also show that the coupling of curvature and confinement nucleates skyrmions at greater thicknesses than those observed for a flat geometry. These findings are particularly important for integrating BPs into miniaturized and curved/flexible devices, including flexible displays, wearable sensors, and smart fabrics.


Merkel cells in mouse skin: intermediate filament pattern, localization, and hair cycle-dependent density.
The distribution and antigen expression of Merkel cells in mouse skin is as yet ill defined. Since the mouse offers an excellent model for studying the origin and functions of Merkel cells, the Merkel cell distribution as well as the expression of intermediate filament proteins and neuronal markers was characterized in C57 BL/6 mouse skin by immunohistochemistry and electron microscopy. Merkel cells in whisker pads, back, and foot pad skin as identified by staining for neuron-specific enolase-an established neuroendocrine marker--expressed cytokeratins (CK) 8,18, and 20 (i.e., simple-epithelial CKs), but not CKs 4 and 13. Sequential double staining for neuron-specific enolase and CK 20 showed consistent co-expression in Merkel cells, establishing CK 20 as a specific immunocytochemical marker for mouse Merkel cells. The Merkel cells also were immunoreactive for synaptophysin but not for neurofilament proteins, peripherin, S-100 protein, and neural cell adhesion molecule. Using CK 8, 18, and 20 as markers, we detected many Merkel cells in the outer roots sheath of vibrissae hair follicles and in foot pad skin. However, only few Merkel cells were found in back skin. These were restricted to small clusters, localized basally within the Haarscheiben epidermis of tylotrich hair follicles, and formed close contacts to prominent nerve fiber terminals as shown by electron microscopy. In striking contrast to human skin, Merkel cells were never found in the epithelium of pelage hair follicles. Even more strikingly, the density of Haarscheiben-associated Merkel cells changed substantially during the highly synchronized, depilation-induced C 57 BL/6 hair cycle, with a minimum in back skin with all hair follicles in telogen or catagen, and a maximum in back skin with all hair follicles in anagen IV-VI. These observations on the Merkel cell hair cycle-dependent distribution in murine skin point to important differences in Merkel cell functions between humans and mice, and raise intriguing questions as to the role of Merkel cells in hair biology.


Positive Cytosolic 5-Nucleotidase 1A Antibodies in Motor Neuron Disease.
Inclusion body myositis (IBM) is the most common acquired myopathy in adults older than 50 years. Muscle biopsy remains the gold standard for diagnosis. Recently described serum antibodies against cytosolic 5-nucleotidase 1A (cN1A) are considered highly specific for IBM. However, positive cN1A antibodies in diseases other than IBM are recently reported. We review 2 cases in which serum antibodies were positive but ancillary testing revealed motor neuron disease. A 68-year-old man presented with asymmetric quadriceps and handgrip weakness prompting concern for IBM. However, electromyography showed purely chronic neurogenic abnormalities, and muscle biopsy was consistent with post-polio syndrome. A 60-year-old woman reported a history of progressive muscle weakness. Despite positive antibodies, examination and electromyography were indicative of amyotrophic lateral sclerosis. Serum cN1A antibodies are not 100% specific for the diagnosis of IBM. Careful clinical, electrophysiologic, and histopathologic correlation is required in workup of individuals with neuromuscular weakness and positive antibodies.


Cross-reactivity of amylin with calcitonin-gene-related peptide binding sites in rat liver and skeletal muscle membranes.
This study examines whether the high degree of sequence identity between amylin and calcitonin-gene-related peptide (CGRP) is reflected in their cross-reactivity at the level of membrane receptor binding. Rat liver plasma membranes contain a specific saturable binding site for 125I-labelled human CGRP-1. Binding reached equilibrium within 30 min and was rapidly reversed by re-incubating membranes in the presence of 1 microM human CGRP. In addition, the presence of 50 mM- or 500 mM-NaCl lowered specific binding by 30% and 77% respectively. Scatchard analysis was consistent with a single high-affinity site with a dissociation constant (Kd) of 0.125 nM and binding capacity (Bmax.) of 580 fmol/mg of membrane protein. Specific binding of 125I-labelled human CGRP-1 to both liver and skeletal muscle membranes was inhibited by human CGRP-1 [IC50 (concn. causing half-maximal inhibition of binding) 0.1-0.3 nM], and rat amylin (IC50 10 nM), but not by human calcitonin. Covalent cross-linking of 125I-CGRP to its binding site in rat skeletal muscle and liver membranes resulted in labelling of a major species of about 70 kDa under reducing conditions and about 55 kDa under alkylating conditions, as visualized on SDS/PAGE. These radiolabelled species were absent in the presence of CGRP or amylin at 1 microM. These results are indicative of a common binding site for both CGRP and amylin in liver and skeletal muscle, and it is suggested that both peptides mediate their actions through the same effector system. The normal physiological importance and the relevance to the pathology of type 2 diabetes of these data are discussed.


Early sensitivity of left perisylvian cortex to relationality in nouns and verbs.
The ability to track the relationality of concepts, i.e., their capacity to encode a relationship between entities, is one of the core semantic abilities humans possess. In language processing, we systematically leverage this ability when computing verbal argument structure, in order to link participants to the events they participate in. Previous work has converged on a large region of left posterior perisylvian cortex as a locus for such processing, but the wide range of experimental stimuli and manipulations has yielded an unclear picture of the region's exact role(s). Importantly, there is a tendency for effects of relationality in single-word studies to localize to posterior temporo-parietal cortex, while argument structure effects in sentences appear in left superior temporal cortex. To characterize these sensitivities, we designed two MEG experiments that cross the factors relationality and eventivity. The first used minimal noun phrases and tested for an effect of semantic composition, while the second employed full sentences and a manipulation of grammatical category. The former identified a region of the left inferior parietal lobe sensitive to relationality, but not eventivity or combination, beginning at 170ms. The latter revealed a similarly-timed effect of relationality in left mid-superior temporal cortex, independent of eventivity and category. The results suggest that i) multiple sub-regions of perisylvian cortex are sensitive to the relationality carried by concepts even in the absence of arguments, ii) linguistic context modulates the locus of this sensitivity, consistent with prior studies, and iii) relationality information is accessed early - before 200ms - regardless of the concept's event status or syntactic category.


Hollow Au nanoflower substrates for identification and discrimination of the differentiation of bone marrow mesenchymal stem cells by surface-enhanced Raman spectroscopy.
Bone marrow mesenchymal stem cells (BMSCs) are multipotent stem cells, which play an important role in the repair of bone injury, angiogenesis, immune diseases, cancer invasion and metastasis. Therefore, increasing attention has been focused on the study of BMSCs. However, the identification and discrimination of the undifferentiated and differentiated BMSCs, which are closely related and morphologically similar, are still a challenge using traditional methods. In this study, we report a novel surface-enhanced Raman scattering (SERS) substrate based on hollow gold nanoflower (HAuNF)-decorated silicon wafers for distinguishing the differentiation of BMSCs. The flower-like, hollow Au nanoparticles were successfully synthesized by a seed-mediated growth approach. We demonstrated that fabricated HAuNF substrates had very good reproducibility, homogeneous SERS activity and a high SERS effect. Using HAuNF substrates as a high-performance in vitro sensing platform allowed us to monitor the changes in the cellular biochemical composition during the differentiation of BMSCs. SERS spectra were analyzed using principal component analysis (PCA), which successfully segregated the subtypes of BMSCs. Furthermore, the results of biological assay suggested that the adipogenic inductor and the osteogenic inductor could induce the differentiation of BMSCs into adipocytes and osteocytes, respectively. The SERS technique based on HAuNF substrates provides a sensitive, efficient and non-invasive detection method for studying the differentiation of stem cells.


p146 type II alveolar epithelial cell antigen is identical to aminopeptidase N.
A prominent membrane protein of rat type II alveolar cells, p146, was originally identified by one of many mouse monoclonal antibodies that were produced to rat lung cells in the course of a search for differentiation antigens that might prove useful in studying lung differentiation. We report here results from analysis of the primary structure of this molecule and, based on this knowledge, the elucidation of the function of the protein. Amino acid sequencing of the NH2-terminal portion of the p146 protein, plus partial sequencing of several peptides obtained by limited proteolysis, indicates it is identical to aminopeptidase N. Further, the immunoaffinity purified p146 protein has aminopeptidase N activity. The discussion includes references to other molecules such as CD 13 and CD 10 (CALLA) that were recognized as differentiation antigens and subsequently found to be peptidases. The possible biological implications of such a peptidase on the luminal surface of type II alveolar cells are also considered.


Factors associated with gay men's sexual behaviours and risk on holiday.
Findings are presented from a questionnaire survey of 562 gay men resident in southern England on their sexual behaviour in the context of holidays. Data were gathered during summer 1996 by means of a self-completion questionnaire; 395 men reported at least one holiday undertaken in 1996 and the analysis focuses on the first or only holiday described. Of 391 men giving details of sexual activity, 187 reported sex with a new partner, 113 men reported penetrative sex and 17 reported penetration without consistent use of condoms. Sexual activity on holiday was predicted by: being on holiday alone or with friends, taking condoms, being motivated by 'gay social life and sex' in planning a holiday, higher expectations of sexual activity/risk and a higher number of new partners at home. Penetrative sex was predicted by: more sexual partners on holiday, taking condoms and higher expectations of sexual activity/risk. Unprotected sex was predicted by: not taking condoms, higher expectations of sexual activity/risk and positive HIV status. Survey findings are discussed in relation to previous research, and implications for travel-related HIV prevention initiatives targeting gay men are explored.


Effect of shrinkage strain, modulus, and instrument compliance on polymerization shrinkage stress of light-cured composites during the initial curing stage.
The aim of this study was to investigate the influence of shrinkage strain, modulus, and instrument compliance on the polymerization shrinkage stress measurement of light-cured composites, and to determine whether the silorane-based low-shrinkage composite shows a low-polymerization shrinkage stress. A universal hybrid; Z250 (Z2), a flowable; Z350 (Z3), and a silorane-based; P90 (P9) composite was examined. A modified "bonded disc method" was used to measure the axial shrinkage strain of the composite. For the measurement of the initial modulus development of composites during light curing, a dynamic oscillatory shear test was undertaken using a custom-made oscillation rheometer. A frequency of 6 Hz and strain amplitude of 0.0091 rad for 20 s was employed and the complex shear modulus (G(*)) was determined. A newly designed stress-strain analyzer was made to measure the shrinkage stress of the composites with two modes: (1) high compliance, or (2) low compliance. The shift between the two modes was controlled by an On-Off switch of a negative feedback circuit. Theoretical shrinkage stress was calculated from the shrinkage strain and modulus measured above, and compared with experimentally measured stress. Data were analyzed with one-way ANOVA and Tukey's post hoc test (alpha=0.05), and correlation analysis was done to investigate the relationship between measured stress and shrinkage strain, modulus, and theoretical stress. The shrinkage strain of Z3 (4.12%) at 10 min was the highest, followed by Z2 (2.31%) and P9 (0.77%). At 10 s after light curing, Z2 showed the highest modulus (466.2 MPa), Z3 (154.1 MPa), and P9 the lowest (130.7 MPa). The measured stresses with low compliance were much higher than those with high compliance. With high compliance, the contraction stress of Z3 was the highest (2.75 MPa), followed by Z2 (1.54 MPa) and P9 (0.48 MPa). In low-compliance mode, the stresses of Z3 (7.93 MPa) and Z2 (7.48 MPa) were similar (p=0.323) while the stress of P9 (3.23 MPa) was much lower. A strong correlation was observed between the theoretical stress and the measured stress with low compliance (R=0.996). In high-compliance mode, the shrinkage strain also showed a near-linear relationship with the stress measured (R=0.937), but the modulus showed a low correlation with the measured stress (R=0.398). Depending on the instrument compliance, polymerization shrinkage stress showed significant differences for each material. In high-compliance shrinkage strain played a greater role, while in low-compliance shrinkage strain and elastic modulus contributed comparably in determining the shrinkage stress. The low-shrinkage silorane-based composite demonstrated considerable reduction in shrinkage strain and stress.


Loss of sirtuin 4 leads to elevated glucose- and leucine-stimulated insulin levels and accelerated age-induced insulin resistance in multiple murine genetic backgrounds.
Several inherited metabolic disorders are associated with an accumulation of reactive acyl-CoA metabolites that can non-enzymatically react with lysine residues to modify proteins. While the role of acetylation is well-studied, the pathophysiological relevance of more recently discovered acyl modifications, including those found in inherited metabolic disorders, warrants further investigation. We recently showed that sirtuin 4 (SIRT4) removes glutaryl, 3-hydroxy-3-methylglutaryl, 3-methylglutaryl, and 3-methylglutaconyl modifications from lysine residues. Thus, we used SIRT4 knockout mice, which can accumulate these novel post-translational modifications, as a model to investigate their physiological relevance. Since SIRT4 is localized to mitochondria and previous reports have shown SIRT4 influences metabolism, we thoroughly characterized glucose and lipid metabolism in male and female SIRT4KO mice across different genetic backgrounds. While only minor perturbations in overall lipid metabolism were observed, we found SIRT4KO mice consistently had elevated glucose- and leucine-stimulated insulin levels in vivo and developed accelerated age-induced insulin resistance. Importantly, elevated leucine-stimulated insulin levels in SIRT4KO mice were dependent upon genetic background since SIRT4KO mice on a C57BL/6NJ genetic background had elevated leucine-stimulated insulin levels but not SIRT4KO mice on the C57BL/6J background. Taken together, the data suggest that accumulation of acyl modifications on proteins in inherited metabolic disorders may contribute to the overall metabolic dysfunction seen in these patients.


Reperfusion arrhythmias: are they only a marker of epicardial reperfusion or continuing myocardial ischemia after acute myocardial infarction?
Reperfusion arrhythmias are associated with epicardial reperfusion but may also be a sign of vascular reperfusion injury which can be seen as no-reflow phenomenon on coronary angiography and predicts in-hospital complications and recovery of left ventricular (LV) function. No-reflow phenomenon (thrombolysis in myocardial infarction [TIMI] <or=2 flow) is frequently observed in patients after mechanical or medical reperfusion procedures for acute myocardial infarction (AMI). The authors hypothesized that reperfusion arrhythmias (or peri-infarct arrhythmias) may be related to continuing myocardial ischemia. They documented all arrhythmia episodes in patients with AMI and compared arrhythmia rates in different therapy groups. They also compared arrhythmia rates according to TIMI flow achieved and those after MI. The highest arrhythmia rate was detected in patients to whom thrombolytic therapy was given for AMI (64%). The arrhythmia rate was lower in patients with primary PCI performed for AMI (46.2%) than in those receiving thrombolytic therapy. The arrhythmia rates according to therapy modalities for AMI were significantly different (p < 0.01). The achieved mean TIMI flow with primary PCI (2.46 +/-0.21 ) was higher than the mean flow achieved after thrombolytic therapy (2.12 +/-0.16). When compared to the arrhythmia rate according to TIMI flow, it was shown that the lowest arrhythmia rate was found in patients with TIMI 3 flow (17.2%) achieved with any procedure after AMI. The arrhythmia rate was 84% in patients with TIMI 2 flow and 33.3% with TIMI 0-1 flow (p <0.001). The arrhythmia rate was appreciably lower after 48 hours of MI. This finding suggests that the continuing myocardial ischemia represented by TIMI flow at the coronary angiography after acute myocardial infarction may have an important role in the pathogenesis of reperfusion arrhythmias.


Does Prenatal Stress Shape Postnatal Resilience? - An Epigenome-Wide Study on Violence and Mental Health in Humans.
Stress during pregnancy widely associates with epigenetic changes and psychiatric problems during childhood. Animal studies, however, show that under specific postnatal conditions prenatal stress may have other, less detrimental consequences for the offspring. Here, we studied mental health and epigenome-wide DNA methylation in saliva following intimate partner violence (IPV) during pregnancy in São Gonçalo, a Brazilian city with high levels of violence. Not surprisingly, mothers exposed to pregnancy IPV expressed elevated depression, PTSD and anxiety symptoms. Children had similar psychiatric problems when they experienced maternal IPV after being born. More surprisingly, when maternal IPV occurred both during (prenatal) and after pregnancy these problems were absent. Following prenatal IPV, genomic sites in genes encoding the glucocorticoid receptor (NR3C1) and its repressor FKBP51 (FKBP5) were among the most differentially methylated and indicated an enhanced ability to terminate hormonal stress responses in prenatally stressed children. These children also showed more DNA methylation in heterochromatin-like regions, which previously has been associated with stress/disease resilience. A similar relationship was seen in prenatally stressed middle-eastern refugees of the same age as the São Gonçalo children but exposed to postnatal war-related violence. While our study is limited in location and sample size, it provides novel insights on how prenatal stress may epigenetically shape resilience in humans, possibly through interactions with the postnatal environment. This translates animal findings and emphasizes the importance to account for population differences when studying how early life gene-environment interactions affects mental health.


The endosymbiont (Buchnera sp.) of the aphid Diuraphis noxia contains plasmids consisting of trpEG and tandem repeats of trpEG pseudogenes.
Most aphids are dependent for their survival on prokaryotic endosymbionts assigned to the genus Buchnera. Among the functions of Buchnera species is the synthesis of tryptophan, which is required by the aphid host. In Buchnera species from the aphid Diuraphis noxia, the genes for anthranilate synthase (trpEG) were found on a plasmid which consisted of seven tandem repeats of a 3.2-kb unit and one 2.6-kb unit which differed in containing a 0.6-kb deletion. One of the 3.2-kb units contained open reading frames corresponding to trpEG; the remaining units contained trpEG pseudogenes (psi). The nucleotide sequence upstream of trpE contained a region that has characteristics of an origin of replication (ori). Relative to trpB (a chromosomal gene), there were about two copies of the trpEG-containing plasmid. Comparisons of the nucleotide sequences of the 3.2-kb units containing trpEG and psi trpEG indicated that most changes occurred in a 700-nucleotide segment that included the region upstream of trpE and the portion of this gene coding for the N terminus. The consequence of these changes was the silencing of trpEG by inactivation of the putative promoter region and premature termination of the TrpE peptide. In contrast, the nucleotide sequence of the segment corresponding to ori was conserved in the units containing trpEG and psi trpEG. We offer a number of speculations on the evolutionary pressure in this lineage which resulted in the silencing of most of trpEG while still retaining the regions resembling ori.


Electrostatics and the membrane association of Src: theory and experiment.
The binding of Src to phospholipid membranes requires both hydrophobic insertion of its myristate into the hydrocarbon interior of the membrane and nonspecific electrostatic interaction of its N-terminal cluster of basic residues with acidic phospholipids. We provide a theoretical description of the electrostatic partitioning of Src onto phospholipid membranes. Specifically, we use molecular models to represent a nonmyristoylated peptide corresponding to residues 2-19 of Src [nonmyr-Src(2-19); GSSKSKPKDPSQRRRSLE-NH2] and a phospholipid bilayer, calculate the electrostatic interaction by solving the nonlinear Poisson-Boltzmann equation, and predict the molar partition coefficient using statistical thermodynamics. The theoretical predictions agree with experimental data obtained by measuring the partitioning of nonmyr-Src(2-19) onto phospholipid vesicles: membrane binding increases as the mole percent of acidic lipid in the vesicles is increased, the ionic strength of the solution is decreased, or the net positive charge of the peptide is increased. The theoretical model also correctly predicts the measured partitioning of the myristoylated peptide, myr-Src(2-19); for example, adding 33% acidic lipid to electrically neutral vesicles increases the partitioning of myr-Src(2-19) 100-fold. Phosphorylating either serine 12 (by protein kinase C) or serine 17 (by cAMP-dependent protein kinase) decreases the partitioning of myr-Src(2-19) onto vesicles containing acidic lipid 10-fold. We investigated the effect of phosphorylation on the localization of Src to biological membranes by expressing fusion constructs of Src's N terminus with a soluble carrier protein in COS-1 cells; phosphorylation produces a small shift in the distribution of the Src chimeras from the plasma membrane to the cytosol.


Induction of whole-body gene deletion via R26-regulated tamoxifen-inducible Cre recombinase activity.
Germline deletion of certain genes causes embryonic lethality, therefore, understanding the effect of deletion of such genes on mammalian pathophysiology remains challenging. Tamoxifen (TAM)-inducible Cre recombinase is widely used for tissue-specific and temporal induction of gene deletion in mice. However, the tamoxifen treatment regimen for the generation of whole-body deletion of a gene is not yet fully standardized for the majority of organs/tissues. Accordingly, we employed GtROSA26 (R26) promoter-regulated Cre and a reporter gene expression strategy. GtROSA26 (R26) is an ubiquitous promoter and mice carrying the R26Cre-ERT2 transgene express Cre-ERT2 in all the cells. Similarly, mice carrying the R26mTOM-mEGFP transgene express mTOM (membrane-targeted tdTomato), in the absence of Cre or mEGFP (membrane-targeted enhanced green fluorescent protein), in the presence of Cre, in all the cells. The progeny carrying one allele of both transgenes were subjected to different TAM regimens, i.e., IP injections (4 injections; 1.35 mg/injection), diet (400 mg TAM-citrate/kg food), or diet (400 mg TAM-citrate/kg food) combined with either TAM-oral gavage (4 gavages; 1.35 mg/gavage) or TAM IP injections (4 injections; 1.35 mg/injection) for 2-weeks beginning at postnatal day (PND) 21 and the extent of Cre recombination in different tissues was determined at PND35. Tamoxifen administration resulted in a transient loss of body weight in all the treatment regimens with a relatively slower rate of weight gain in the TAM-diet plus TAM-oral gavage group compared to other groups. While the efficiency of Cre recombination, as determined by the expression of mEGFP protein, was variable among tissues, major tissues such as the liver, heart, lungs, spleen, and thymus-showed almost complete recombination. No recombination was evident in any of the tissues examined from the control mice. In general, the efficiency of Cre recombination was better with a combined regimen of TAM-diet with either TAM-injections or TAM-oral gavage compared to TAM-diet alone or TAM-injections alone. Our results demonstrate that a combination of TAM-diet with either TAM-injections or TAM-oral gavage can be employed for the efficient deletion of a gene in the whole body. Our findings will provide technical expertise to the researchers employing TAM-inducible Cre for the deletion of floxed genes in varied tissues.


Discovery and development of Seliciclib. How systems biology approaches can lead to better drug performance.
Seliciclib (R-Roscovitine) was identified as an inhibitor of CDKs and has undergone drug development and clinical testing as an anticancer agent. In this review, the authors describe the discovery of Seliciclib and give a brief summary of the biology of the CDKs Seliciclib inhibits. An overview of the published in vitro and in vivo work supporting the development as an anti-cancer agent, from in vitro experiments to animal model studies ending with a summary of the clinical trial results and trials underway is presented. In addition some potential non-oncology applications are explored and the potential mode of action of Seliciclib in these areas is described. Finally the authors argue that optimisation of the therapeutic effects of kinase inhibitors such as Seliciclib could be enhanced using a systems biology approach involving mathematical modelling of the molecular pathways regulating cell growth and division.


Cytochrome c oxidase is preferentially synthesized in the rough endoplasmic reticulum--mitochondrion complex in rat liver.
The specific activity and content of cytochrome oxidase in the rough endoplasmic reticulum--mitochondrion complex are higher than in the mitochondrial fraction. Radiolabelling studies with the use of hepatocytes and isolated microsomal and rough endoplasmic reticulum--mitochondrion fractions, followed by immunoprecipitation with anti-(cytochrome oxidase) antibody, reveal that the nuclear-coded cytoplasmic subunits of cytochrome oxidase are preferentially synthesized in the latter fraction. The results have a bearing on the mechanism of transport of these subunits into mitochondria.


Hyponatremia and/or hyperkalemia in patients treated with the standard dose of trimethoprim-sulfamethoxazole.
High-dose trimethoprim-sulfamethoxazole (TMP-SMX) is known to cause hyperkalemia by blocking amiloride-sensitive sodium (Na) channels in distal nephrons. The purpose of this study was to establish whether the standard dose of TMP-SMX could cause electrolyte disorders. Serum Na, potassium (K) and creatinine (Cr) levels were examined retrospectively in 53 of 77 patients prescribed TMP-SMX, before and after taking the antibiotic combination. Electrolyte disorders (Na < 135 mEq/l and/or K > 5.0 mEq/l) were found in 14 of the 53 patients (26.4%) during TMP-SMX treatment. The average dose was 145.7 +/- 24.9 mg/day. The dose of TMP was significantly larger in patients with electrolyte disorders (267.7 +/- 84.2 mg vs. 101.9 +/- 9.38 mg, p = 0.0024). Electrolyte disorders were also seen in 9.1% and 22.2% of patients given the low dose (TMP < 80 mg) or standard dose (TMP 80-120 mg) of TMP-SMX, respectively. Electrolyte disorders were seen in 85.7% of patients with renal dysfunction (Cr > 1.2 mg/dl), compared with 17.5% of patients with normal renal function (p = 0.0008). Logistic regression analysis showed that the dose of TMP and the presence of renal dysfunction increased the incidence of electrolyte disorders with an odds ratio of 2.35 and 80.29, respectively. Electrolyte disorders, particularly hyperkalemia and hyponatremia can be detected in patients given TMP-SMX. These disorders are more frequent in patients given high doses, but can also be detected after low-dose administration. Renal dysfunction accelerates the incidence of electrolyte disorders induced by TMP-SMX.


Ultraearly Intraventricular Hemorrhage Growth Predicts Early Neurologic Deterioration and Poor Functional Outcome After Acute Intracerebral Hemorrhage.
Background The presence of intraventricular hemorrhage (IVH) was extensively investigated and was associated with poor outcome in patients with intracerebral hemorrhage (ICH). However, the effect of the speed of ventricular bleeding on outcomes is unknown. Methods and Results We prospectively included patients with ICH who had baseline computed tomography scans within 6 hours after ictus between January 2016 and October 2021. The clinical characteristics were compared between patients with and without early neurologic deterioration (END). Ultraearly IVH growth (uIVHG) was defined as baseline IVH volume by onset-to-imaging time. The association between uIVHG and outcomes was assessed by using multivariable logistic regression analysis. We established the ultraearly IVH growth (uIVH) score and compared the areas under the receiver operating characteristic curves of the existing scores for predicting END. A total of 299 patients were finally enrolled. Of those, 38 patients (12.7%) experienced END at 24 hours and 89 patients (29.8%) had poor outcomes at 90 days. After adjustment for confounding factors, uIVHG (odds ratio, 1.061 [95% CI, 1.011-1.113]; P=0.016) was independently associated with END in multivariable analysis. A prediction score was developed on the basis of the logistic model. The uIVH score was developed as a sum of individual points (0-6) based on age, hematoma volume, National Institutes of Health Stroke Scale, hematoma expansion, and uIVHG ≥2.5 mL/h. In comparison with the ICH score and modified Emergency Department ICH Scale, the uIVH score exhibited best performance in the prediction of END. Conclusions uIVHG is associated with early neurologic deterioration and poor functional outcome in patients with ICH.


Usefulness of intraoperative electrical subcortical mapping during surgery for low-grade gliomas located within eloquent brain regions: functional results in a consecutive series of 103 patients.
Although a growing number of authors currently advocate surgery to treat low-grade gliomas, controversy still persists, especially because of the risk of inducing neurological sequelae when the tumor is located within eloquent brain areas. Many researchers performing preoperative neurofunctional imaging and intraoperative electrophysiological methods have recently reported on the usefulness of cortical functional mapping. Despite the frequent involvement of subcortical structures by these gliomas, very few investigators have specifically raised the subject of fiber tracking. The authors in this report describe the importance of mapping cortical and subcortical functional regions by using intraoperative real-time direct electrical stimulations during resection of low-grade gliomas. Between 1996 and 2001, 103 patients harboring a corticosubcortical low-grade glioma in an eloquent area, with no or only mild deficit, had undergone surgery during which intraoperative electrical mapping of functional cortical sites and subcortical pathways was performed throughout the procedure. Both eloquent cortical areas and corresponding white fibers were systematically detected and preserved, thus defining the resection boundaries. Despite an 80% rate of immediate postoperative neurological worsening, 94% of patients recovered their preoperative status within 3 months--10% even improved--and then returned to a normal socioprofessional life. Eighty percent of resections were classified as total or subtotal based on control magnetic resonance images. The use of functional mapping of the white matter together with cortical mapping allowed the authors to optimize the benefit/risk ratio of surgery of low-grade glioma invading eloquent regions. Given that preoperative fiber tracking with the aid of neuroimaging is not yet validated, we used intraoperative real-time cortical and subcortical stimulations as a valuable adjunct to the other mapping methods.


Habitat type predicts genetic population differentiation in freshwater invertebrates.
A basic challenge in evolutionary biology is to establish links between ecology and evolution of species. One important link is the habitat template. It has been hypothesized, that the spatial and temporal settings of a habitat strongly influence the evolution of species dispersal propensity. Here, we evaluate the importance of the habitat type on genetic population differentiation of species using freshwater habitats as a model system. Freshwater habitats are either lentic (standing) or lotic (running). On average, lotic habitats are more stable and predictable over space and time than lentic habitats. Therefore, lentic habitats should favour the evolution of higher dispersal propensity which ensures population survival of lentic species. To test for such a relationship, we used extensive data on species' genetic population differentiation of lentic and lotic freshwater invertebrates retrieved from published allozyme studies. Overall, we analysed more than 150 species from all over the world. Controlling for several experimental, biological and geographical confounding effects, we always found that lentic invertebrates exhibit, on average, lower genetic population differentiation than lotic species. This pattern was consistent across insects, crustaceans and molluscs. Our results imply fundamental differences in genetic population differentiation among species adapted to either lentic or lotic habitats. We propose that such differences should occur in a number of other habitat types that differ in spatio-temporal stability. Furthermore, our results highlight the important role of lotic habitats as reservoirs for evolutionary processes and the potential for rapid speciation.


[Ultrastructural findings during human myocardial preservation with HTK solution].
Although the majority of recent scientific advances developed in the field of myocardial preservation have improved surgery procedures, it seems that during cardiothoracic surgery ischemia-reperfusion continues triggering clinical postoperative problems. This report focused on the changes human capillary endothelial cells and nerve endings suffer after cardiopulmonary bypass. The study involved four patients who received Bretschneider solution for cardioplegia during mitral stenosis surgery. Biopsies of right atrium were taken prior to and after CPB to be analyzed under electron microscopy. Samples taken after CPB showed detachment of myocardial capillary basal membrane, endothelial edema, and widespread nerve-ending destruction. Nevertheless, the shelter provided by the most advanced cardioplegic solutions, myocardial endothelial edema, and nerve-ending destruction cannot be completely prevented. Thus, it is possible to suggest that low interstitial concentration of Na(+), which activates Na(+)/Ca(++) exchanger, and inflow of Ca(++), present during ischemia-reperfusion, may lead to endothelial and neuronal cell damage, triggering cardiac contraction dysfunction. This mechanism would explain at least in part some of the problems patients face during the postoperative state.


The neuronal repellent SLIT2 is a target for repression by EZH2 in prostate cancer.
The neuronal repellent SLIT2 is repressed in a number of cancer types primarily through promoter hypermethylation. SLIT2, however, has not been studied in prostate cancer. Through genome-wide location analysis we identified SLIT2 as a target of polycomb group (PcG) protein EZH2. The EZH2-containing polycomb repressive complexes bound to the SLIT2 promoter inhibiting its expression. SLIT2 was downregulated in a majority of metastatic prostate tumors, showing a negative correlation with EZH2. This repressed expression could be restored by methylation inhibitors or EZH2-suppressing compounds. In addition, a low level of SLIT2 expression was associated with aggressive prostate, breast and lung cancers. Functional assays showed that SLIT2 inhibited prostate cancer cell proliferation and invasion. Thus, this study showed for the first time the epigenetic silencing of SLIT2 in prostate tumors, and supported SLIT2 as a potential biomarker for aggressive solid tumors. Importantly, PcG-mediated repression may serve as a precursor for the silencing of SLIT2 by DNA methylation in cancer.


Integration of absolute cardiovascular disease risk assessment into routine blood cholesterol testing at pathology services.
Absolute cardiovascular disease (CVD) risk assessment is recommended for primary prevention of CVD, yet uptake in general practice is limited. Cholesterol requests at pathology services provide an opportunity to improve uptake by integrating absolute CVD risk assessment with this service. This study aimed to assess the feasibility of such an additional service. Two-hundred and ninety-nine patients (45-74 years) referred to pathology services for blood cholesterol had measurement of all variables required to determine absolute CVD risk according to Framingham calculator (blood pressure, age, sex, smoking and diabetes status via self-report). Data were recorded via computer-based application. The absolute risk score was communicated via the report sent to the referring medical practitioner as per usual practice. Evaluation questionnaires were completed immediately post visit and at 1-, 3- and 6-month follow-up via telephone (n = 262). Absolute CVD risk reports were issued for 90% of patients. Most patients (95%) reported that the length of time for the pathology service assessment was acceptable, and 91% that the self-directed computer-based application was easy to use. Seventy-eight per cent reported a preference for pathology services to conduct absolute CVD risk assessment. Only 2% preferred a medical practitioner. Of follow-up patients, 202 (75%) had a consultation with a medical practitioner, during which, aspects of CVD risk prevention were discussed (cholesterol and blood pressure 74% and 69% of the time, respectively). Measurement of absolute CVD risk in pathology services is feasible, highly acceptable among middle-to-older adults and may increase uptake of guideline-directed care in general practice.


The guanosine binding mechanism of the Tetrahymena group I intron.
The Tetrahymena group I intron catalyzes self-splicing through two consecutive transesterification reactions, using a single guanosine-binding site (GBS). In this study, we constructed a model RNA that contains the GBS and a conserved guanosine nucleotide at the 3'-terminus of the intron (omegaG). We determined by NMR the solution structure of this model RNA, and revealed the guanosine binding mechanism of the group I intron. The G22 residue, corresponding to omegaG, participates in a base triple, G22 xx G3 x C12, hydrogen-bonding to the major groove edge of the Watson-Crick G3 x C12 pair. The G22 residue also interacts with A2, which is semi-conserved in all sequenced group I introns.


Aptorchis glandularis n. sp. (Digenea: Plagiorchioidea) from the northwestern red-faced turtle, Emydura australis, (Pleurodira: Chelidae) in the Kimberley, Western Australia.
Aptorchis glandularis n. sp. is described from the intestine of the northwestern red-faced turtle, Emydura australis (Pleurodira: Chelidae), in the Kimberley, Western Australia. This digenean is morphologically most similar to Aptorchis aequalis but can be differentiated readily from the latter species by the presence of ventral glands arranged in 3 rows. This feature is unique among plagiorhioidean digeneans and resembles the glands observed in some monostome digeneans in Notocotylidae and Microscaphidiidae. Comparison of approximately 2,600 bases of ribosomal DNA (partial 18S, complete ITS1+5.8S+ITS2, partial 28S), obtained from all 5 known Aptorchis species, strongly supports the status of Aptorchis glandularis n. sp. as a new species. Phylogenetic analysis of these sequence data demonstrates 2 strongly supported clades (A. pearsoni + A. megacetabulus) and (A. aequalis + A. glandularis n. sp.), with A. megapharynx representing a separate lineage. This is the first report of an endoparasite from Emydura australis.


Characteristics of GABA receptors on the ocellar L-neurons of American cockroach Periplaneta americana.
The ocellar L-neurons of cockroach Periplaneta americana were used in the present study as model systems to investigate the pharmacological properties of the GABA receptors. To do so, a glass microelectrode was impaled into the axon of the L-neurons to record the membrane potential intracellularly and to monitor membrane response to GABA treatment and cercal stimulation by air puff. The traditional GABA and their receptor agonists were introduced through perfusion and/or iontophoresis to monitor their effects on the L-neurons. The GABA receptor antagonists were administered by perfusion to examine if the response of the L-neurons to GABA and/or cercal stimulation was changed. The results revealed that administration of GABA, muscimol and imidazole acetic acid, two GABAA agonists, produced depolarization on the L-neurons. However, treatment of 3-APS and guanidine acetic acid, another two GABAA agonists, evoked hyperpolarization on the L-neurons. Among those tested antagonists, only picrotoxin, GABAA antagonist, antagonize the depolarization induced by GABA and/or cercal stimulation. More interestingly, administration of strychnine, glycine receptor antagonist, largely attenuated the depolarization response of the L-neurons to cercal stimulation. This attenuation caused by strychnine was even stronger than that initiated by varied GABA antagonists. In addition, phaclofen, a GABAB receptor antagonist, showed no antagonistic effect. These results strongly suggest that the characteristics of GABA receptors of the ocellar L-neurons may differ from those in vertebrates. It may be more likely to be a novel GABA receptor.


Paradoxical activation of an inwardly rectifying potassium channel mutant by spermine: "(b)locking" open the bundle crossing gate.
Intracellular polyamines are endogenous blockers of inwardly rectifying potassium (Kir) channels and underlie steeply voltage-dependent rectification. Kir channels with strong polyamine sensitivity typically carry a negatively charged side chain at a conserved inner cavity position, although acidic residues at any pore-lining position in the inner cavity are sufficient to confer polyamine block. We have identified unique consequences of a glutamate substitution in the region of the helix bundle crossing of Kir6.2. Firstly, glutamate substitution at Kir6.2 residue F168 generates channels with intrinsic inward rectification that does not require blockade by intracellular polyamines or Mg(2+). In addition, these F168E channels exhibit a unique "spiked" tail phenotype, whereby large decaying inward tail currents are elicited upon spermine unbinding. This contrasts with the time-dependent recovery of current typically associated with blocker unbinding from ion channels. Interestingly, Kir6.2[F168E] channels exhibit a paradoxical biphasic conductance-voltage relationship in the presence of certain polyamines. This reflects channel blockade at positive voltages, channel stimulation at intermediate voltages, and exclusion of spermine from the pore at negative voltages. These features are recapitulated by a simple kinetic scheme in which weakly voltage-dependent spermine binding to a "shallow" site in the pore (presumably formed by the introduced glutamate at F168E) stabilizes opening of the bundle crossing gate. These findings illustrate the potential for dichotomous effects of a blocker in a long pore (with multiple binding sites), and offer a unique example of targeted modulation of the Kir channel gating apparatus.


Interrogating the Role of Receptor-Mediated Mechanisms: Biological Fate of Peptide-Functionalized Radiolabeled Gold Nanoparticles in Tumor Mice.
To get a better insight on the transport mechanism of peptide-conjugated nanoparticles to tumors, we performed in vivo biological studies of bombesin (BBN) peptide functionalized gold nanoparticles (AuNPs) in human prostate tumor bearing mice. Initially, we sought to compare AuNPs with thiol derivatives of acyclic and macrocyclic chelators of DTPA and DOTA types. The DTPA derivatives were unable to provide a stable coordination of (67)Ga, and therefore, the functionalization with the BBN analogues was pursued for the DOTA-containing AuNPs. The DOTA-coated AuNPs were functionalized with BBN[7-14] using a unidentate cysteine group or a bidentate thioctic group to attach the peptide. AuNPs functionalized with thioctic-BBN displayed the highest in vitro cellular internalization (≈ 25%, 15 min) in gastrin releasing peptide (GRP) receptor expressing cancer cells. However, these results fail to translate to in vivo tumor uptake. Biodistribution studies following intravenous (IV) and intraperitoneal (IP) administration of nanoconjugates in tumor bearing mice indicated that the presence of BBN influences to some degree the biological profile of the nanoconstructs. For IV administration, the receptor-mediated pathway appears to be outweighed by the EPR effect. By contrast, in IP administration, it is reasoned that the GRPr-mediated mechanism plays a role in pancreas uptake.


Genetic risk factors for cerebrovascular disease in children with sickle cell disease: design of a case-control association study and genomewide screen.
The phenotypic heterogeneity of sickle cell disease is likely the result of multiple genetic factors and their interaction with the sickle mutation. High transcranial doppler (TCD) velocities define a subgroup of children with sickle cell disease who are at increased risk for developing ischemic stroke. The genetic factors leading to the development of a high TCD velocity (i.e. cerebrovascular disease) and ultimately to stroke are not well characterized. We have designed a case-control association study to elucidate the role of genetic polymorphisms as risk factors for cerebrovascular disease as measured by a high TCD velocity in children with sickle cell disease. The study will consist of two parts: a candidate gene study and a genomewide screen and will be performed in 230 cases and 400 controls. Cases will include 130 patients (TCD > or = 200 cm/s) randomized in the Stroke Prevention Trial in Sickle Cell Anemia (STOP) study as well as 100 other patients found to have high TCD in STOP II screening. Four hundred sickle cell disease patients with a normal TCD velocity (TCD < 170 cm/s) will be controls. The candidate gene study will involve the analysis of 28 genetic polymorphisms in 20 candidate genes. The polymorphisms include mutations in coagulation factor genes (Factor V, Prothrombin, Fibrinogen, Factor VII, Factor XIII, PAI-1), platelet activation/function (GpIIb/IIIa, GpIb IX-V, GpIa/IIa), vascular reactivity (ACE), endothelial cell function (MTHFR, thrombomodulin, VCAM-1, E-Selectin, L-Selectin, P-Selectin, ICAM-1), inflammation (TNFalpha), lipid metabolism (Apo A1, Apo E), and cell adhesion (VCAM-1, E-Selectin, L-Selectin, P-Selectin, ICAM-1). We will perform a genomewide screen of validated single nucleotide polymorphisms (SNPs) in pooled DNA samples from 230 cases and 400 controls to study the possible association of additional polymorphisms with the high-risk phenotype. High-throughput SNP genotyping will be performed through MALDI-TOF technology using Sequenom's MassARRAY system. It is expected that this study will yield important information on genetic risk factors for the cerebrovascular disease phenotype in sickle cell disease by clarifying the role of candidate genes in the development of high TCD. The genomewide screen for a large number of SNPs may uncover the association of novel polymorphisms with cerebrovascular disease and stroke in sickle cell disease.


Design and applications of three dimensional covalent organic frameworks.
Covalent organic frameworks (COFs), as an emerging class of crystalline porous polymers connected by dynamic covalent bonds, have been well studied over the past decade. Recently, three dimensional (3D) COFs have attracted extensive interest for the synthesis and applications of novel COFs. The principal reason for this rising trend is based on their unique porous features and excellent performances compared to previously reported two dimensional (2D) frameworks with the layered AA-stacking mode. This critical review describes the current state-of-the-art development of 3D COFs in the design principles, synthetic methods, functionalization strategies, and potential applications. Some major challenges associated with future perspectives are further discussed, inspiring the development of 3D COFs.


[Percutaneous puncture of the pancreas controlled by computerized tomography].
The method of puncture biopsy under the control of computer tomography was used in 29 patients with diseases of the pancrease. Correct diagnosis was obtained in 27 patients. In 13 patients aimed biopsy was associated with the aspiration of cysts (in 7 patients) and drainage (in 6 patients). No complications of punctures of the pancreas under the control of computer tomography were noted.


cGAS-STING signalling regulates microglial chemotaxis in genome instability.
Defective DNA damage signalling and repair is a hallmark of age-related and genetic neurodegenerative disease. One mechanism implicated in disease progression is DNA damage-driven neuroinflammation, which is largely mediated by tissue-resident immune cells, microglia. Here, we utilise human microglia-like cell models of persistent DNA damage and ATM kinase deficiency to investigate how genome instability shapes microglial function. We demonstrate that upon DNA damage the cytosolic DNA sensing cGAS-STING axis drives chronic inflammation and a robust chemokine response, exemplified by production of CCL5 and CXCL10. Transcriptomic analyses revealed that cell migratory pathways were highly enriched upon IFN-β treatment of human iPSC-derived microglia, indicating that the chemokine response to DNA damage mirrors type I interferon signalling. Furthermore, we find that STING deletion leads to a defect in microglial chemotaxis under basal conditions and upon ATM kinase loss. Overall, this work provides mechanistic insights into cGAS-STING-dependent neuroinflammatory mechanisms and consequences of genome instability in the central nervous system.


Vitrase ISTA Pharmaceuticals.
Vitrase is under development by ISTA Pharmaceuticals Inc (formerly Advanced Corneal Systems, ACS) and Otsuka Pharmaceutical (in Japan) for the potential treatment of vitreous hemorrhage and diabetic retinopathy. In November 1998, it was approved in Mexico for the clearing of vitreous hemorrhage [305542]. In October 1998, Vitrase was designated a Fast-Track product for the vitreous hemorrhage indication by the FDA [303024], [307576]. The initial section of an NDA was filed in January 2002, which comprised the non-clinical pharmacology and toxicology sections [434925]. In April 2002, following discussions with the FDA, ISTA reported that it would be proceeding with the preparation and submission of the clinical and other required sections of the NDA for Vitrase for the treatment of vitreous hemorrhage [447148]. In February 2002, analysts at CIBC World Markets Corp predicted Vitrase would be launched in early 2003, with worldwide sales reaching 200 million dollars within 5 years [444945].


[Lyme borreliosis].
Lyme disease is the most common of tickborne spirochetoses in France. Over fifty years, several clinical forms have been identified. Following the description of the disease in the United States, links were recognised between the forms presenting infectious, cutaneous, neurological and rheumatic symptoms. Although its epidemiology has now been well established, this spirochetosis can still pose difficult diagnostic problems despite serological investigations. Concerning therapy, rapid treatment is highly valuable. The prescription of ceftriaxone is now highly recommended.


Measuring the accuracy of ICA-based artifact removal from TMS-evoked potentials.
The analysis and interpretation of transcranial magnetic stimulation (TMS)-evoked potentials (TEPs) relies on successful cleaning of the artifacts, which typically mask the early (0-30 ms) TEPs. Independent component analysis (ICA) is possibly the single most utilized methodology to clean these signals. ICA-based cleaning is reliable provided that the input data are composed of independent components. Differently, in case the underlying components are to some extent dependent, ICA algorithms may yield erroneous estimates of the components, resulting in incorrectly cleaned data. We aim to ascertain whether TEP signals are suited for ICA. We present a systematic analysis of how the properties of simulated artifacts imposed on measured artifact-free TEPs affect the ICA results. The variability of the artifact waveform over the recorded trials is varied from deterministic to stochastic. We measure the accuracy of ICA-based cleaning for each level of variability. Our findings indicate that, when the trial-to-trial variability of an artifact component is small, which can result in dependencies between underlying components, ICA-based cleaning biases towards eliminating also non-artifactual TEP data. We also show that the variability can be measured using the ICA-derived components, which in turn allows us to estimate the cleaning accuracy. As TEP artifacts tend to have small trial-to-trial variability, one should be aware of the possibility of eliminating brain-derived EEG when applying ICA-based cleaning strategies. In practice, after ICA, the artifact component variability can measured, and it predicts to some extent the cleaning reliability, even when not knowing the clean ground-truth data.


Carbonic anhydrase II and IV mRNA in rabbit nephron segments: stimulation during metabolic acidosis.
Carbonic anhydrase (CA) facilitates renal bicarbonate reabsorption and acid excretion. Cytosolic CA II catalyzes the buffering of intracellular hydroxyl ions by CO2, whereas membrane-bound CA IV catalyzes the dehydration of carbonic acid generated from the secretion of protons. Although CA II and IV are expressed in rabbit kidney, it is not entirely clear which segments express which isoforms. It was the purpose of this study to characterize the expression of CA II and CA IV mRNAs by specific segments of the nephron using semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) and to determine the effect of chronic metabolic acidosis on CA expression by those segments. Individual nephron segments (usually 1-2 mm) were isolated by microdissection and subjected to RT-PCR. Amplification was performed simultaneously for CA IV, CA II, and malate dehydrogenase (MDH), a housekeeping gene. The intensities of the PCR products were quantitated by densitometry. CA IV mRNA was expressed by S1 and S2 proximal tubules and by outer medullary collecting duct from inner stripe (OMCDi) and outer stripe and initial inner medullary collecting duct (IMCDi). CA II mRNA was expressed by S1, S2, and S3 proximal tubules, thin descending limb, connecting segment (CNT), and all collecting duct segments. Acid loading induced CA IV mRNA expression in S1 and S2 proximal tubules and in OMCDi and IMCDi. CA II mRNA was induced by acidosis in all three proximal segments and nearly all distal segments beginning with CNT. No upregulation of MDH mRNA expression occurred. These adaptive increases in CA II and IV mRNAs are potentially important in the kidney's adaptation to chronic metabolic acidosis.


Incidence of lower urinary tract injury at the time of total laparoscopic hysterectomy.
To determine the incidence of and risk factors for injury to the lower urinary tract during total laparoscopic hysterectomy. All patients who underwent total laparoscopic hysterectomy for benign disease from January 1, 2002 to December 31, 2005, at an academic medical center are included. Subjects undergoing laparoscopic-assisted vaginal hysterectomy, supracervical hysterectomy, or hysterectomy for malignancy were excluded. Intraoperative cystoscopy with intravenous indigo carmine was routinely performed. Relevant data were abstracted to determine the incidence of lower urinary tract injury, predictors of injury, and postoperative complications. Total laparoscopic hysterectomy was performed in 126 consecutive subjects. Two (1.6%) cystotomies were noted and repaired before cystoscopy was performed. Two (1.6%) additional cystotomies were detected during cystoscopy. Absent ureteral spill of indigo carmine was detected in 2 subjects: 1 (0.8%) with previously unknown renal disease and 1 (0.8%) with ureteral obstruction that was relieved with subsequent suture removal. Only 40% (2/5) of injuries were recognized without the use of cystoscopy with indigo carmine. The overall incidence of injury to the lower urinary tract was 4.0%. No subjects required postoperative intervention to the lower urinary tract within the 6-week perioperative period. Performing a ureterolysis was associated with an increased rate (odds ratio 8.7, 95%CI, 1.2-170, P=0.024) of lower urinary tract injury. Surgeons should consider performing cystoscopy with intravenous indigo carmine dye at the time of total laparoscopic hysterectomy.


NIR initiated and pH sensitive single-wall carbon nanotubes for doxorubicin intracellular delivery.
In this work, we synthesized a pH sensitive poly(ethylene glycol)-doxorubicin (PEG-DOX) on single-wall carbon nanotubes (SWNTs). Using multimodal nonlinear optical imaging microscopy, we found that low power (1 mW cm-2) near-infrared radiation can initiate prodrug burst release from the carbon nanotubes in seconds. The successful SWNTs capping with PEG-DOX (denoted PEG-DOX@SWNT) was determined by transmission electron microscopy and FTIR results. The in vitro release of DOX from the PEG-DOX@SWNT was evaluated upon changes of pH values and NIR treating time. The cytotoxicity of the PEG-DOX@SWNT was also evaluated. This dual-sensitive delivery system based on SWNTs provides a facile approach to promote drug release and kill cancer cells.


Diversity and seasonal fluctuation of predominant microbial communities in Bhitarkanika, a tropical mangrove ecosystem in India.
Different groups of microorganisms are present in mangrove areas, and they perform complex interactions for nutrient and ecological balances. Since little is known about microbial populations in mangroves, this study analyzed the microbial community structure and function in relation to soil physico-chemical properties in Bhitarkanika, a tropical mangrove ecosystem in India. Spatial and seasonal fluctuations of thirteen important groups of microorganisms were evaluated from the mangrove forest sediments during different seasons, along with soil physico-chemical parameters. The overall microbial load (x10(5)cfu/g soil) in soil declined in the order of heterotrophic, free living N2 fixing, Gram-negative nitrifying, sulphur oxidizing, Gram-positive, spore forming, denitrifying, anaerobic, phosphate solubilizing, cellulose degrading bacteria, fungi and actinomycetes. Populations of the heterotrophic, phosphate solubilizing, sulphur oxidizing bacteria and fungi were more represented in the rainy season, while, Gram-negative, Gram-positive, nitrifying, denitrifying, cellulose decomposing bacteria and actinomycetes in the winter season. The pool size of most of other microbes either declined or maintained throughout the season. Soil nutrients such as N, P, K (Kg/ha) and total C (%) contents were higher in the rainy season and they did not follow any common trend of changes throughout the study period. Soil pH and salinity (mS/cm) varied from 6-8 and 6.4-19.5, respectively, and they normally affected the microbial population dynamics. Determination of bacterial diversity in Bhitarkanika mangrove soil by culture method showed the predominance of bacterial genera such as Bacillus, Pseudomonas, Desulfotomaculum, Desulfovibrio, Desulfomonas, Methylococcus, Vibrio, Micrococcus, Klebsiella and Azotobacter. Principal component analysis (PCA) revealed a correlation among local environmental variables with the sampling locations on the microbial community in the mangrove soil.


The pathology and causes of tissue eosinophilia in the gastrointestinal tract.
Eosinophilic inflammation in the gastrointestinal tract may occur as a primary eosinophilic disorder or as a secondary response with other causes. Primary eosinophilic gastrointestinal disorders (EGIDs) are Th2-mediated allergic diseases that overlap pathogenetically with atopic conditions involving other organs. The pathological diagnosis of primary EGIDs can be challenging, as the quantity of eosinophils considered to be 'abnormal' is difficult to define, and the diagnosis, by definition, requires exclusion of the far more common secondary causes. Our understanding of the basic biology and natural history of eosinophilic oesophagitis has advanced considerably over the last decade, whereas other EGIDs have proven more difficult to characterize; nonetheless, some recent advances have been made. This review summarizes current knowledge regarding the clinical presentation, diagnosis, natural history and treatment of EGIDs, including eosinophilic oesophagitis. We also draw attention to the numerous secondary causes of tissue eosinophilia in the gastrointestinal tract, and suggest a practical approach to the histological assessment, diagnosis and reporting of EGIDs.


Predictive factors for post-total thyroidectomy hypocalcaemia.
Post total thyroidectomy hypocalcemia is a frequent and serious complication of thyroid surgery. Predictive Factors of its risk, justifying the prescription of preventive treatment are to be determined. To identify the predictive factors of hypocalcemia secondary to total thyroidectomy. We report a retrospective study involving 260 patients operated with total thyroidectomy at the ENT department of the military hospital of Tunis over a period of 5 years. A statistical study was carried out to find out predictors of hypocalcemia. Our patients were women in 85% of cases with an average age of 49 years. The incidence of postoperative hypocalcemia was 31.9%. Hypoparathyroidism was definitive in 1.1% of cases. Univariate statistical analysis identified the following factors: presence of cervical swelling at the initial physical exam, gradual onset of mass, thyroid nodule size greater than 4 cm, preoperative calcium level <2.25 mmol/L, one-shot thyroidectomy operating time, histological appearance of thyroiditis and accidental excision of a parathyroid gland. The factors retained in the multivariate study were: gradual onset of mass, nodule size> 4 cm, preoperative calcium level <2.25 mmol/L and histological appearance of thyroiditis. Post total thyroidectomy hypocalcemia is linked to intrinsic factors of the thyroid, preoperative calcium level, preoperative and intraoperative parathyroid lesions. Its prevention would include: supplementation of patients with hyperthyroidism or with low preoperative calcium levels, systematic identification of the parathyroid glands and their reimplantation in cases of devascularization.


Diagnostic capacity of biomechanical indices from a dynamic bidirectional applanation device in pellucid marginal degeneration.
To evaluate the diagnostic capacity of the keratoconus match index (KMI) and keratoconus match probability (KMP) classification from a dynamic bidirectional applanation device (Ocular Response Analyzer) in eyes with pellucid marginal degeneration (PMD). Department of Ophthalmology, University Clinics Saarland, Homburg, Germany. Cross-sectional study. Pellucid marginal degeneration eligibility was confirmed by inferior peripheral thinning, corneal protrusion, and irregular astigmatism. The KMI and KMP parameters in PMD eyes (study group) were compared with those in normal eyes (control group). The KMI's overall predictive accuracy was assessed operating characteristic (ROC) curves. The relationship between KMI and corneal hysteresis (CH), the corneal resistance factor (CRF), and a series of Scheimpflug camera indices was evaluated with Spearman analysis (r). The mean KMI in the study group (40 eyes) and control group (40 eyes) was 0.34 ± 0.43 (SD) and 0.95 ± 0.30, respectively (P<.001). The KMI correlated significantly with the CH, CRF, and most Scheimpflug camera indices. Regression analysis indicated that the index of height decentration (r = -0.877, P<.001) was the primary determinant of the KMI. Moreover, the KMP index identified 50.0%, 29.16%, and 20.83% of PMD eyes as ectatic, suspect for ectasia, and normal, respectively. The ROC curve analysis of the KMI parameter indicated a predictive accuracy of 94.8% (cutoff point 0.626; sensitivity 85.71%; specificity 90.1%). The KMI seems to be a promising diagnostic index for PMD. In contrast, the KMP index identified a significant percentage of topographically defined PMD eyes as normal, limiting its diagnostic value in PMD. No author has a financial or proprietary interest in any material or method mentioned.


Belatacept Combined With Transient Calcineurin Inhibitor Therapy Prevents Rejection and Promotes Improved Long-Term Renal Allograft Function.
Belatacept, a T cell costimulation blocker, demonstrated superior renal function, lower cardiovascular risk, and improved graft and patient survival in renal transplant recipients. Despite the potential benefits, adoption of belatacept has been limited in part due to concerns regarding higher rates and grades of acute rejection in clinical trials. Since July 2011, we have utilized belatacept-based immunosuppression regimens in clinical practice. In this retrospective analysis of 745 patients undergoing renal transplantation at our center, we compared patients treated with belatacept (n = 535) with a historical cohort receiving a tacrolimus-based protocol (n = 205). Patient and graft survival were equivalent for all groups. An increased rate of acute rejection was observed in an initial cohort treated with a protocol similar to the low-intensity regimen from the BENEFIT trial versus the historical tacrolimus group (50.5% vs. 20.5%). The addition of a transient course of tacrolimus reduced rejection rates to acceptable levels (16%). Treatment with belatacept was associated with superior estimated GFR (belatacept 63.8 mL/min vs. tacrolimus 46.2 mL/min at 4 years, p < 0.0001). There were no differences in serious infections including rates of cytomegalovirus or BK viremia. We describe the development of a costimulatory blockade-based strategy that ultimately allows renal transplant recipients to achieve calcineurin inhibitor-free immunosuppression.


Disseminated Cryptococcosis with Marked Eosinophilia in a Postpartum Woman.
Disseminated cryptococcosis usually develops in immunosuppressed patients. A 33-year-old postpartum woman developed disseminated cryptococcosis with marked eosinophilia. She presented with a cough and a week-long fever. A computed tomography scan showed multiple pulmonary nodules randomly distributed. Eosinophils were observed to have increased in number in both her peripheral blood and bronchoalveolar lavage fluid. A transbronchial lung biopsy and cerebrospinal fluid specimens revealed findings consistent with cryptococcal infection. Disseminated cryptococcosis can present with marked eosinophilia of the peripheral blood and lung tissues. Additionally, the postpartum immune status may sometimes be involved in the development of opportunistic infections in previously healthy women.


Recent advances in nanocarrier-loaded gels: Which drug delivery technologies against which diseases?
The combination of pharmaceutical technologies can be a wise choice for developing innovative therapeutic strategies. The association of nanocarriers and gels provides new therapeutic possibilities due to the combined properties of the two technologies. Gels support the nanocarriers, localize their administration to the target tissue, and sustain their release. In addition to the properties afforded by the gel, nanocarriers can provide additional drug sustained release or different pharmacokinetic and biodistribution profiles than those from nanocarriers administered by the conventional route to improve the drug therapeutic index. This review focuses on recent (over the last ten years) in vivo data showing the advances and advantages of using nanocarrier-loaded gels. Liposomes, micelles, liquid and solid lipid nanocapsules, polymeric nanoparticles, dendrimers, and fullerenes are all nanotechnologies which have been recently assessed for medical applications, such as cancer therapy, the treatment of cutaneous and infectious diseases, anesthesia, the administration of antidepressants, and the treatment of unexpected diseases, such as alopecia.


The human cytoplasmic dynein interactome reveals novel activators of motility.
In human cells, cytoplasmic dynein-1 is essential for long-distance transport of many cargos, including organelles, RNAs, proteins, and viruses, towards microtubule minus ends. To understand how a single motor achieves cargo specificity, we identified the human dynein interactome by attaching a promiscuous biotin ligase ('BioID') to seven components of the dynein machinery, including a subunit of the essential cofactor dynactin. This method reported spatial information about the large cytosolic dynein/dynactin complex in living cells. To achieve maximal motile activity and to bind its cargos, human dynein/dynactin requires 'activators', of which only five have been described. We developed methods to identify new activators in our BioID data, and discovered that ninein and ninein-like are a new family of dynein activators. Analysis of the protein interactomes for six activators, including ninein and ninein-like, suggests that each dynein activator has multiple cargos.


The novel AKT inhibitor afuresertib shows favorable safety, pharmacokinetics, and clinical activity in multiple myeloma.
The PI3K/AKT pathway is constitutively active in hematologic malignancies, providing proliferative and antiapoptotic signals and possibly contributing to drug resistance. We conducted an open-label phase 1 study to evaluate the maximum tolerated dose (MTD), safety, pharmacokinetics, and clinical activity of afuresertib-an oral AKT inhibitor-in patients with advanced hematologic malignancies. Seventy-three patients were treated at doses ranging from 25 to 150 mg per day. The MTD was established at 125 mg per day because of 2 dose-limiting toxicities in the 150-mg cohort (liver function test abnormalities). The most frequent adverse events were nausea (35.6%), diarrhea (32.9%), and dyspepsia (24.7%). Maximum plasma concentrations and area under the plasma concentration-time curves from time 0 to 24 hours were generally dose proportional at > 75-mg doses; the median time to peak plasma concentrations was 1.5 to 2.5 hours post dose, with a half-life of approximately 1.7 days. Three multiple myeloma patients attained partial responses; an additional 3 attained minimal responses. Clinical activity was also observed in non-Hodgkin lymphoma, Langerhan's cell histiocytosis, and Hodgkin disease. Single-agent afuresertib showed a favorable safety profile and demonstrated clinical activity against hematologic malignancies, including multiple myeloma.