7T MPFLAIR versus MP2RAGE for Quantifying Lesion Volume in Multiple Sclerosis.
Use of fluid-attenuated inversion recovery (FLAIR) scans to quantify multiple sclerosis (MS) lesion volume on 7 Tesla (7T) magnetic resonance imaging (MRI) has many downsides, including poor image homogeneity. There are little data about the relative benefit of alternative modalities. The purpose of this paper is to investigate if magnetization-prepared 2 rapid acquisition gradient echo (MP2RAGE) is a viable alternative to FLAIR for robust lesion volume measurement and disability correlations. Forty-seven participants with MS underwent annual brain 7T MRIs. Magnetization-prepared FLAIR (MPFLAIR) and MP2RAGE (both at .7 mm3 isotropic resolution) sequences from a total of 80 MRI scans from 47 subjects were reviewed. White matter lesion (WML) masks were manually constructed from MPFLAIR and T1 maps (from MP2RAGE). Lesion volumes (normalized to intracranial volume) were compared to clinical characteristics and disability scales scores by Pearson or Spearman correlation, as appropriate. Relative correlation strength was compared by Fisher r- to z-transformation. Normalized lesion volume was greater in MPFLAIR masks (median .005 [range, .001-.030]) than from T1 maps (median .003 [range, .000-.015]). However, lesion volumes between MPFLAIR and T1 maps were highly correlated (rho = .87, P < .001). WML masks from both modalities correlated with most disability measures with no significant difference in the strength of correlation. 7T MPFLAIR and MP2RAGE T1 map-based WML volumes are highly intercorrelated and both correlate with disability. Thus, MP2RAGE may be a viable alternative to FLAIR-based methods for WML measurement on 7T MRI in MS research.


Estimating heritability of refractive error in Koreans: the Korea National Health and Nutrition Examination Survey.
To estimate the familial correlation and heritability of refractive error in general Korean population. From the Korea National Health and Nutrition Examination Survey, 13 258 subjects of 7920 families, who were aged ≥19 years, were included in the study. Using variance components analysis, the additive genetic effect, or heritability, and the common and unique environmental effects on refractive error were examined, adopting common environments shared by cohabiting family or by siblings. The proportions of hyperopia, myopia and high myopia in Koreans were 0.8%, 45.2% and 5.7% respectively. The correlation coefficients of spherical equivalent (SE) were 0.257 for parent-offspring pairs, 0.410 for sibling pairs and 0.112 for spouse pairs (p < 0.001 for all). Common environment shared by siblings affected the variation of SE significantly (p < 0.001), but that shared by cohabitants did not (p = 0.395). Adopting common environment shared by siblings, the heritability, common environmental effect and unique environmental effect of refractive error were 42.1 ± 3.3%, 11.8 ± 3.5% and 46.1 ± 3.9% respectively. Heritabilities of hyperopia, myopia and high myopia were 45.7%, 44.3% and 68.9% respectively. Adjusted odds ratios of myopia among offspring were 3.78 given one parent has myopia and 4.43 when both parents have myopia. Refractive error is influenced by common environment shared by siblings. The heritability of refractive error is higher for high myopia than for myopia or hyperopia.


[Local ice application in therapy of kinetic limb ataxia. Clinical assessment of positive treatment effects in patients with multiple sclerosis].
Upper limb ataxia is one of the most disabling symptoms of patients with multiple sclerosis (MS). There are some clinically tested therapeutic strategies, especially with regard to cerebellar tremor. But most of the methods used for treatment of limb ataxia in physiotherapy and occupational therapy are not systematically evaluated, e.g. the effect of local ice applications, as reported by MS patients and therapists, respectively. We investigated 21 MS patients before and in several steps 1 up to 45 min after cooling the most affected forearm. We used a series of 6 tests, including parts of neurological status and activities of daily living as well. At each step skin temperature and nerve conduction velocity were recorded. All tests were documented by video for later offline analysis. Standardized evaluation was done by the investigators and separately by an independent second team, both of them using numeric scales for quality of performance. After local cooling all patients showed a positive effect, especially a reduction of intentional tremor. In most cases this effect lasted 45 min, in some patients even longer. We presume that a decrease in the proprioceptive afferent inflow-induced by cooling-may be the probable cause of this reduction of cerebellar tremor. Patients can use ice applications as a method of treating themselves when a short-time reduction of intention tremor is required, e.g. for typing, signing or self-catheterization.


A review on the impact of 4-quinolones on the normal oropharyngeal and intestinal human microflora.
During the last few years the impact of the newer 4-quinolones, ciprofloxacin, enoxacin, norfloxacin, ofloxacin and pefloxacin, on the human microflora has been studied by several investigators. This review article summarizes the published data concerning these studies. The results show that the oropharyngeal flora is affected only slightly or not at all by the 4-quinolones. All the newer 4-quinolones have a similar effect on the normal intestinal flora. The gram-negative aerobic flora is strongly suppressed during administration of 4-quinolones, while the gram-positive flora is only slightly affected. The anaerobic microflora is hardly affected at all. The emergence of resistant bacterial strains is uncommon, although one study shows increased MIC-values for anaerobes during ciprofloxacin administration. Replacement by yeasts or other inherently resistant microorganisms does not often seem to be a problem. High concentrations of the 4-quinolones are reached in faeces, values between 100-2,200 mg/kg being reported. Since the 4-quinolones do not cause marked ecological disturbances in the intestinal microflora, they may be suitable for selective decontamination in immunocompromised patients, for prophylaxis of urinary tract infections and for treatment of bacterial intestinal infections.


Infection with prevalent clinical strains of Mycobacterium tuberculosis leads to differential maturation of monocyte derived dendritic cells.
The link between innate and adaptive host immune response to Mycobacterium tuberculosis (M.tb) is driven by dendritic cells (DC). In this study, we examined the ability of prevalent clinical strains from south India (S7, S10) and laboratory strain H37Rv (Rv) to induce maturation of monocyte derived dendritic cells (MoDC). The phenotypic and functional changes of DC upon infection with different strains of M.tb were evaluated. It was observed that S7 and Rv strains partially hampered the maturation of MoDC as reflected by the low expression of maturation markers and co-stimulatory markers when compared to LPS stimulated MoDC. In contrast, strain S10 infected DC showed a marked increase in the expression of these markers. The functional property was investigated by the ability of infected MoDC to induce T-cell proliferation and to stimulate secretion of IFN-gamma by CD4(+)T-cells. It was found that Rv and S7 infected MoDC were less efficient in inducing T-cell proliferation. The secretion of IL-12 by Rv and S7 infected MoDC was also found to be significantly lesser than LPS stimulated MoDC. On the other hand, S10 infected MoDC showed enhanced T-cell stimulation and cytokine secretion. Together these results indicate that there is a substantial variability in the capacity of M.tb clinical strains to induce maturation of DC which may be dependent upon their virulence.


A facile and general route to synthesize silica-coated SERS tags with the enhanced signal intensity.
Silica-coated SERS tags have been attracting greater attention in recent years. However, the reported methods to synthesize these tags are tedious, and often subjected to the limited signal intensity. Here, we report a facile and general method to prepare the silica-coated Ag SERS tags with the enhanced signal intensity by no introducing the primers. This approach mainly depends on the colloidal stability of the Ag NPs in alcohol solution. By decreasing the concentration of salt in Ag NP solution, the citrate-stabilized Ag NPs can be well dispersed in alcohol solution. Based on this, the Ag SERS tags can be directly coated with thickness-controlled and homogeneous silica shells. This approach is highly reproducible for silica shell growth and signal intensity, not depending on the properties of Raman molecules, proved by 7 kinds of the Raman molecules. Moreover, this kind of SERS tags coated with silica hold the stronger SERS signals than the traditional method due to no interference from the priming molecules.


A general system to integrate lacZ fusions into the chromosomes of gram-negative eubacteria: regulation of the Pm promoter of the TOL plasmid studied with all controlling elements in monocopy.
A new procedure is described to recombine plasmid-borne lacZ fusions into the chromosome of gram-negative eubacteria in order to study promoter activity in monocopy. The procedure is based upon the insertion into the chromosome of a target bacterium of a recombinant transposon that carries DNA sequence homology to the regions flanking lacZ fusions present in multicopy promotor-probe vectors, which can be mobilized via RP4-mediated transfer but are unable to replicate in non-enteric bacteria. Double recombination between the promoter-probe vectors and the chromosomal homology region of the transposon is genetically selected by reconstruction and expression of wild-type sequences from truncated lacZ and aadA (streptomycin/spectinomycin) resistance genes in the homology fragment and from an amber mutation carrying lacZ and aadA genes present in the plasmid vectors. The structure of desired clones is confirmed by screening for loss of the transposon-encoded kanamycin resistance marker. We have used this procedure to assemble in monocopy in Pseudomonas putida the regulatory elements controlling expression of the XylS-activated Pm promoter of the TOL catabolic plasmid pWWO. We show here that the Pm promoter undergoes a XylS-independent, strictly growth-phase-controlled activation by benzoate but not meta-toluate. In the presence of XylS, however, activation by both effectors involves a combination of growth phase-dependent and -independent controls.


The intracellular organization of actin and tubulin in cultured C-1300 mouse neuroblastoma cells (clone NB41A3).
Cells of clone NB41A3 of the C-1300 mouse neuroblastoma were grown to a critical density at which many of the cells flatten, assume a variety of shapes and sizes and some sprout processes resembling neurites. We have studied the distribution of actin and tubulin in these cells using fluorescence microscopy and antibodies against actin or tubulin under these conditions. Actin-containing structures are variably arranged and predominantly associated with motile areas of the cell periphery including the growth cone. Microtubules appear to run radially from the perinuclear area towards the cell periphery. When neurites are present, microtubules converge into them and run to the growth cone but rarely contact its edge.


Farnesylthiosalicylic acid-derivatized PEI-based nanocomplex for improved tumor vaccination.
Cancer vaccines that make use of tumor antigens represent a promising therapeutic strategy by stimulating immune responses against tumors to generate long-term anti-tumor immunity. However, vaccines have shown limited clinical efficacy due to inefficient delivery. In this study, we focus on vaccine delivery assisted by nanocomplexes for cancer immunotherapy. Nanocomplex-mediated vaccination can efficiently deliver nucleic acids encoding neoantigens to lymphoid tissues and antigen-presenting cells. Polyethylenimine (PEI) was conjugated with farnesylthiosalicylic acid (FTS) to form micelles. Subsequent interaction with nucleic acids led to formation of polymer/nucleic acid nanocomplexes of well-controlled structure. Tumor transfection via FTS-PEI was much more effective than that by PEI, other PEI derivatives, or naked DNA. Significant numbers of transfected cells were also observed in draining lymph nodes (LNs). In vivo delivery of ovalbumin (OVA; a model antigen) expression plasmid (pOVA) by FTS-PEI led to a significant growth inhibition of the OVA-expressing B16 tumor through presentation of OVA epitopes as well as other epitopes via epitope spreading. Moreover, in vivo delivery of an endogenous melanoma neoantigen tyrosinase-related protein 2 (Trp2) also led to substantial tumor growth inhibition. FTS-PEI represents a promising transfection agent for effective gene delivery to tumors and LNs to mediate effective neoantigen vaccination.


Intracranial myiasis by Hypoderma bovis (Linnaeus) in a horse.
Acute neurologic disease associated with intracranial migration of a first instar larva of a warble fly, Hypoderma bovis (Linnaeus), was observed in a 14-year-old Quarter Horse gelding in western Montana. The disease was characterized by incoordination of gait, circling to the left, head tilt to the right, partial paralysis of the right side of the face, and impaired vision in the right eye. Two and one-half hours after it was first noticed sick, the horse collapsed and was euthanized. Massive hemorrhage unaccompanied by necrosis or significant cellular response was present in the right side of the midbrain and pons.


Magnetic in situ determination of surface coordination motifs by utilizing the degree of particle agglomeration.
Most analytical techniques used to study the surface chemical properties of superparamagnetic iron oxide nanoparticles (SPIONs) are barely suitable for in situ investigations in liquids, where SPIONs are mostly applied for hyperthermia therapy, diagnostic biosensing, magnetic particle imaging or water purification. Magnetic particle spectroscopy (MPS) can resolve changes in magnetic interactions of SPIONs within seconds at ambient conditions. Herein, we show that by adding mono- and divalent cations to citric acid capped SPIONs, the degree of agglomeration can be utilized to study the selectivity of cations towards surface coordination motifs via MPS. A favored chelate agent, like ethylenediaminetetraacetic acid (EDTA) for divalent cations, removes cations from coordination sites on the SPION surface and causes redispersion of agglomerates. The magnetic determination thereof represents what we call a "magnetically indicated complexometric titration". The relevance of agglomerate sizes for the MPS signal response is studied on a model system of SPIONs and the surfactant cetrimonium bromide (CTAB). Analytical ultracentrifugation (AUC) and cryogenic transmission electron microscopy (cryo-TEM) reveal that large micron-sized agglomerates are required to significantly change the MPS signal response. With this work, a fast and easy-to-use characterization method to determine surface coordination motifs of magnetic nanoparticles in optically dense media is demonstrated.


Prolactin receptors in primary cultures of carcinogen-induced rat-mammary tumors.
7,12-Dimethylbenz[alpha]anthracene-induced rat mammary tumors were dissociated with collagenase and hyaluronidase and placed into primary culture. In most cultures, specific binding of 125I-labeled ovine prolactin was (i) lower than that for the original tumors unless bovine prolactin (1 microgram/ml) had been added to the dissociation medium, and (ii) varied with the type of growth medium used. The level of prolactin binding in cultured cells was relatively constant for the first 7-10 days. Prolactin binding in cultured cell homogenates was maximal at pH 7.0, proportional to cell protein, specific for prolactin, and reached a steady state by 12 h at 22 degrees C. The half-maximum inhibition of 125I-labeled prolactin binding by unlabeled prolactin was 100 ng/ml for cells grown in 5-1000 ng of prolactin/ml. After prolactin was removed from the growth medium, the level of available binding sites progressively increased, reached a maximum at 48 h and then declined. At 48 h, the dissociation constant for prolactin binding (Kd approximately 1 x 10(-10) M) was comparable to that in tumors. In some cultured tumors, a 48-h treatment with 0.5 or 1.0 ng of prolactin/ml caused an apparent increase in the level of prolactin binding. Prolactin increased DNA synthesis and its removal caused a reduction in [3H]estradiol and [3H]-R5020 binding to cultured cell cytosols.


Effect of detergent binding on cytochrome P450 2B4 structure as analyzed by X-ray crystallography and deuterium-exchange mass spectrometry.
Multiple crystal structures of CYP2B4 have demonstrated the binding of the detergent 5-cyclohexyl-1-pentyl-β-D-maltoside (CYMAL-5) in a peripheral pocket located adjacent to the active site. To explore the consequences of detergent binding, X-ray crystal structures of the peripheral pocket mutant CYP2B4 F202W were solved in the presence of hexaethylene glycol monooctyl ether (C8E6) and CYMAL-5. The structure in the presence of CYMAL-5 illustrated a closed conformation indistinguishable from the previously solved wild-type. In contrast, the F202W structure in the presence of C8E6 revealed a detergent molecule that coordinated the heme-iron and extended to the protein surface through the substrate access channel 2f. Despite the overall structural similarity of these detergent complexes, remarkable differences were observed in the A, A', and H helices, the F-G cassette, the C-D and β4 loop region. Hydrogen-deuterium exchange mass spectrometry (DXMS) was employed to probe these differences and to test the effect of detergents in solution. The presence of either detergent increased the H/D exchange rate across the plastic regions, and the results obtained by DXMS in solution were consistent in general with the relevant structural snapshots. The study provides insight into effect of detergent binding and the interpretation of associated conformational dynamics of CYP2B4.


Overview of cartilage biology and new trends in cartilage stimulation.
This article reviews the basics of articular cartilage biology, which provide a necessary foundation for understanding the evolving field of articular cartilage injury and repair. The currently popular treatment options for osteochondral injury (microfracture, osteochondral autograft transfer system, osteochondral allograft, autologous chondrocyte implantation, and the use of scaffolds with autologous chondrocyte implantation) document the significant advances made in this area in the past 2 decades. Integration of newly available information and technology derived from advances in molecular biology and tissue engineering holds even greater promise for continued advances in optimal management of this challenging problem.


No social distancing between ORAI channels.
ORAI1 is established as an essential component of Ca2+ release-activated Ca2+ (CRAC) channel which mediates store-operated Ca2+ entry (SOCE). However, the contributions of ORAI2 and ORAI3 to SOCE are not understood. We highlight a recent study which shows that ORAI proteins form heteromeric channels which tune SOCE over a range of stimulus intensities.


Issues in dental and surgical management of the pregnant patient.
The "echo boom" of the 1980s has increased the incidence of pregnancy in our patient population. When a gravid patient has an urgent or emergent problem, knowledge of the unique aspects of pregnancy is necessary for optimal patient care. Maternal health is protected by an appreciation of the physiologic changes and demands of pregnancy. Fetal well-being is protected by avoidance of teratogens, fetal anoxia, and premature labor. Attention to these factors allows the clinician to provide treatment with minimal maternal or fetal risk.


Plasma lecithin: cholesterol acyltransferase activity in liver disease.
The plasma lecithin: cholesterol acyltransferase was determined in patients with various liver diseases and the relationship between this enzyme activity and the other liver function tests were studied including long term observations. Lecithin: cholesterol acyltransferase activity in fulminant hepatitis and liver cirrhosis showed a significant decrease in comparison with normal volunteers. Although the enzyme activity of hepatoma showed significant decrease, they were ascribed to the influence of concomitant liver cirrhosis. The enzyme activity showed insignificant changes in the acute and chronic hepatitis and alcoholic liver disease. Lecithin: cholesterol acyltransferase activity was correlated with the concentration of cholesterolester rather than with the ratio of esters to cholesterol. In addition, it was well correlated with pseudocholine esterase and serum albumin. The lecithin: cholesterol acyltransferase activity in the cases during follow-up period varied in good parallel with cholesterol-esters concentration and pseudocholine esterase in the cases with acute hepatitis; with serum albumin in the cases with liver cirrhosis. Furthermore, it varied inversely with SGPT in the cases with acute hepatitis. In a case with hepatoma, lecithin: cholesterol acyltransferase activity decreased more sharply than the cholesterolesters concentration and serum albumin immediately before death.


Calcium-channel blocking agents for gastrointestinal disorders.
Calcium-channel blocking agents have a potential role in regulation of gastrointestinal tract function by decreasing smooth muscle contraction and possibly inhibiting cellular secretion. Most studies to date have concentrated on the ability of these drugs to inhibit smooth muscle contraction. Verapamil and diltiazem have been shown in animals (opossum, baboon) to produce decreased contractions in esophageal smooth muscle, resulting in a decreased amplitude of peristalsis and decreased lower esophageal sphincter pressures. In studies in man, oral doses of diltiazem and nifedipine have likewise been shown to have similar effects on the esophagus. At present, there is no experimental evidence of a major antisecretory effect of these drugs. In clinical trials, nifedipine has been shown to have a greater effect than placebo in improving symptoms in patients with achalasia, and diltiazem has been suggested as potential therapy in patients with chest pain secondary to excessive esophageal contraction. The precise role for calcium-channel blocking drugs in therapy of gastrointestinal disease is still being explored.


Multicenter evaluation of the RAPIDEC® CARBA NP assay for the detection of carbapenemase production in clinical isolates of Enterobacterales and Pseudomonas aeruginosa.
Carbapenem-resistant Gram-negative bacilli are a major public health problem. Accurate and rapid detection of carbapenemase-producing organisms can facilitate appropriate infection prevention measures. The objective was to evaluate the performance of the RAPIDEC® CARBA NP assay (RAPIDEC), a screening assay that utilizes a pH indicator to detect carbapenem hydrolysis within 2 h. A multicenter study evaluated 306 clinical bacterial strains of Enterobacterales (n = 257) and Pseudomonas aeruginosa (n = 49). The RAPIDEC was compared to a composite reference standard-the Clinical Laboratory Standards Institute (CLSI) Carba NP assay, PCR for specific carbapenemase genes (blaKPC, blaNDM, blaOXA-48-like, blaVIM and blaIMP), and phenotypic carbapenem susceptibility testing. The assay was evaluated using two culture incubation times for the bacterial isolates: "routine"(cultures incubated 18-24 h) and "short" (cultures incubated 4-5 h). For the routine incubation, the overall percent agreement was 98.7% with a positive percent agreement (PPA) of 99.6% and a negative percent agreement (NPA) of 97.4%; there were five false positives and one false negative. For the short incubation, the overall percent agreement was 98.0% with a PPA of 98.5% and a NPA of 97.3%; there were five false positives and four false negatives. RAPIDEC results for the P. aeruginosa isolates were 100% concordant with the reference standard for both incubation times. The RAPIDEC assay is an accurate and rapid (≤ 2 h) assay for the detection of the most common carbapenemases in clinical isolates. Growth from a short incubation culture may be used to reliably detect carbapenemase production in clinical strains.


Sun exposure is an aggravating factor responsible for the recalcitrant facial erythema in adult patients with atopic dermatitis.
In Japan, a considerable number of adult patients with atopic dermatitis suffer from recalcitrant facial erythema that resists common treatment with topical corticosteroids and antihistamines. Our purpose was to investigate the potential role of sun exposure in the aggravation of these facial lesions. The history of photoaggravation was taken from 74 adult patients with atopic dermatitis who suffered from recalcitrant facial erythema. Repeated UVB and UVA phototests were performed in 36 patients. Surface markers of infiltrating cells in UVB-provoked lesions were characterized immunohistochemically. Forty-one of 74 patients experienced an exacerbation of the facial lesions after sun exposure. UVB testing revealed an abnormal, papular response in 14 of 36 patients. All of the 14 patients complained of clinical aggravation after sun exposure. No abnormal reactions were observed at UVA testing. In UVB-provoked lesions, CD4+ cells were predominant to CD8+ cells. Exposure to UVB radiation may be responsible for the recalcitrant facial erythema in at least some of the patients with atopic dermatitis.


Intraparenchymal renal artery pseudoaneurysm following nephrostomy tube insertion in a patient with a solitary kidney: A case report.
Intraparenchymal renal artery aneurysms are uncommon and represent less than 10% of all renal artery aneurysms. They are caused by trauma or iatrogenic injury, and their rupture can lead to life-threatening hemorrhage. We report the case of a 48-year-old male with history of left solitary kidney and orthotopic neobladder, who presented with massive hematuria 7 days after nephrostomy tube placement because of obstructive uropathy and acute renal failure due to ureteroileal stricture. An abdominal CT angiography revealed an intraparenchymal renal artery aneurysm, and it was successfully treated with superselective endovascular embolization, achieving maximal parenchymal preservation.


Ligand effects on the protein ensemble: unifying the descriptions of ligand binding, local conformational fluctuations, and protein stability.
Detailed description of the structural and physical basis of allostery, cooperativity, and other manifestations of long-range communication between binding sites in proteins remains elusive. Here we describe an ensemble-based structural-thermodynamic model capable of treating explicitly the coupling between ligand binding reactions, local fluctuations in structure, and global conformational transitions. The H(+) binding reactions of staphylococcal nuclease and the effects of pH on its stability were used to illustrate the properties of proteins that can be described quantitatively with this model. Each microstate in the native ensemble was modeled to have dual structural character; some regions were treated as folded and retained the same atomic geometry as in the crystallographic structure while other regions were treated thermodynamically as if they were unfolded. Two sets of pK(a) values were used to describe the affinity of each H(+) binding site. One set, calculated with a standard continuum electrostatics method, describes H(+) binding to sites in folded parts of the protein. A second set of pK(a) values, obtained from model compounds in water, was used to describe H(+) binding to sites in unfolded regions. An empirical free energy function, parameterized to reproduce folding thermodynamics measured by differential scanning calorimetry, was used to calculate the probability of each microstate. The effects of pH on the distribution of microstates were determined by the H(+) binding properties of each microstate. The validity of the calculations was established by comparison with a number of different experimental observables.


Monosyllabic Mandarin tone productions by 3-year-olds growing up in Taiwan and in the United States: interjudge reliability and perceptual results.
The author compared monosyllabic Mandarin lexical tones produced by 3-year-old Mandarin-speaking children growing up in Taiwan and in the United States. Following the procedures in Wong, Schwartz, and Jenkins (2005), the author collected monosyllabic tone productions from 3-year-old Mandarin-speaking children in Taiwan and low-pass filtered them to eliminate lexical information but retain tone information. Five Mandarin-speaking adults residing in Taiwan categorized these filtered tones and those produced by the Mandarin-speaking children growing up in the United States, the latter of which was reported in Wong et al. (2005). Agreements on tone categorization by judges residing in Taiwan and in the United States were evaluated. Tone accuracy of children growing up in Taiwan and the United States were examined and compared. The Mandarin-speaking judges residing in the United States and in Taiwan showed high agreements on tone categorization. None of the 4 tones produced by the Mandarin-speaking children growing up in the United States and in Taiwan was adultlike.Children in Taiwan made more errors in Tone 2 and Tone 4 than did Mandarin-speaking children growing up in the United States. Accuracy rates of Tone 1 and Tone 3 were comparable in the 2 groups of children. Mandarin tone acquisition is a protracted process. Three-year-old Mandarin-speaking children growing up in Taiwan and the United States show similar developmental patterns and have not yet produced monosyllabic tones with adultlike accuracy.


Acute effect of ketotifen on the dose-response curve of histamine and methacholine in asthma.
This study was designed to assess the acute effect of 1 mg of orally administered ketotifen on the dose-response curve to both histamine and methacholine, in 15 patients with bronchial asthma. From this curve we measured the concentration that produced a 20% fall in FEV1 (PC20) and reactivity, defined as the slope of the curve beyond the threshold dose. Results were analysed comparing the change in log PC20 and in reactivity before and after a double-blind administration of ketotifen and placebo. Ketotifen, compared to placebo, significantly increased the histamine PC20 (P less than 0.001) and reduced reactivity (P less than 0.001). After adjusting for starting FEV1 the difference of effect between ketotifen and placebo was still significant, although to a lesser degree (P less than 0.01). In contrast, ketotifen did not modify the methacholine dose-response curve. No significant changes were observed in FEV1 after ketotifen. The results suggest that acute administration of ketotifen has a selective action on histamine-activated pathways of bronchoconstriction.


Are molluscan maximum life spans determined by long-term cycles in benthic communities?
A review of the maximum longevity of bivalves and gastropods indicates that a greater than average number of life spans coincide with the periods of long-term cycles in marine communities. Apparently, long-term cycles exert an important influence on marine communities by affecting the life spans of constituent species. Gastropods and bivalves are affected differently, longevities being determined by some cycles more than others in each group. Overall, molluscan longevities tend to be slightly longer than the corresponding cycle suggesting that there is selection pressure for life spans slightly longer than the cycle controlling recruitment success and generational replacement.


Effects of pressure on vesicle size, density, and distribution in the canine carotid arterial endothelium.
The purpose of the present experiment was to study the effects of internal hydrostatic pressure on vesicle size, density, and distribution in the canine carotid arterial endothelium by transmission electron microscopy. The pressures applied in this study were 0 (control), 40, 60, 80, 100, and 150 mm Hg. The results of transmission electron microscopy and computer analysis on the plasmalemmal vesicles of aortic endothelium showed that luminal, abluminal, and junctional vesicles all increased their diameter as the pressure was raised from 0 mm Hg, reaching a maximum at 80 mm Hg, and then decreased in size with further increases in pressure to 150 mm Hg. There was a significant difference in diameter among vesicles in different regions of the endothelium, with the diameter of luminal vesicles larger than those of abluminal and junctional vesicles. The densities of vesicles showed very little change from 0 to 80 mm Hg; but they increased markedly as the pressure was further raised from 80 to 150 mm Hg. These results indicate that pressure is an important mechanical factor governing the size and density of plasmalemmal vesicles in aortic endothelium.


Mutations in an S4 segment of the adult skeletal muscle sodium channel cause paramyotonia congenita.
The periodic paralyses are a group of autosomal dominant muscle diseases sharing a common feature of episodic paralysis. In one form, paramyotonia congenita (PC), the paralysis usually occurs with muscle cooling. Electrophysiologic studies of muscle from PC patients have revealed temperature-dependent alterations in sodium channel (NaCh) function. This observation led to demonstration of genetic linkage of a skeletal muscle NaCh gene to a PC disease allele. We now report the use of the single-strand conformation polymorphism technique to define alleles specific to PC patients from three families. Sequencing of these alleles defined base pair changes within the same codon, which resulted in two distinct amino acid substitutions for a highly conserved arginine residue in the S4 helix of domain 4 in the adult skeletal muscle NaCh. These data establish the chromosome 17q NaCh locus as the PC gene and represent two mutations causing the distinctive, temperature-sensitive PC phenotype.


Identification and characterization of anti-osteoclastogenic peptides derived from the cytoplasmic tail of receptor activator of nuclear factor kappa B.
Pathological bone resorption by osteoclasts is primarily treated with bisphosphonates. Because the administration of bisphosphonates is associated with a risk for multiple adverse symptoms, a precise understanding of the mechanisms underlying osteoclastogenesis is required to develop drugs with minimal side-effects. Osteoclastogenesis depends on receptor activator of nuclear factor kappa B (RANK) signaling mediated by TRAF6. We previously identified a highly conserved domain in the cytoplasmic tail of RANK (HCR), which did not share any significant homology with other proteins and was essential for osteoclastogenesis. HCR acts as a platform for the formation of Gab2- and Vav3-containing signal complexes, and ectopic expression of the HCR peptide inhibits osteoclastogenesis. Here, we uncover the mechanisms of HCR peptide-mediated inhibition of osteoclastogenesis. Expression of either the amino- or carboxyl-terminal half of the HCR peptide (N- or C-peptide) independently inhibited RANK signaling prior to cell-cell fusion. In contrast, expression of the GY-peptide, which is a part of the C-peptide, did not significantly affect prefusion RANK signaling, but did inhibit cell-cell fusion to prevent formation of multinucleated mature osteoclasts. Moreover, Gab2, which is involved in RANK signaling by binding TRAF6, bound the C-peptide but not the N-peptide, suggesting that the C- and the N-peptides sequester TRAF6 in a Gab2-dependent and Gab2-independent manner, respectively. In contrast, the GY-peptide did not bind Gab2 but could bind Vav3, which mediates signaling for cell-cell fusion. Collectively, we propose that the HCR peptide inhibits osteoclastogenesis through two modes of action-inhibition of (1) prefusion RANK signaling and (2) cell-cell fusion by blocking TRAF6- and Vav3-mediated signaling, respectively.


Regulation of seed number and female incitation of mate competition by a pH-dependent proteinaceous inhibitor of pollen grain germination in Leucaena leucocephala.
Leucaena leucocephala generally produces pods with more than 7-9 seeds. This is regulated by the stigmatic inhibition of pollen grain germination when the pollen grains are less than a critical number in the stigma. This number-dependent inhibition of pollen grain germination is effected by a pH-dependent proteinaceous inhibitor active at the stigmatic pH. Only when the pollen grains in the stigma exceed the critical number, they inactivate the inhibitor by collectively raising the stigmatic pH and thus overcoming the inhibition. The adaptive significance of such pre-fertilization mechanism for the female in inciting mate competition among the pollen grains is discussed. The evolution of en masse pollen grain dispersal units is explained as a sexual selection strategy by males in response to such stigmatic inhibition by females.


Overexpression of the orotate phosphoribosyl-transferase gene enhances the effect of 5-Fluorouracil in head and neck squamous cell carcinoma in vitro.
5-Fluorouracil (5-FU) is a widely used drug in head and neck squamous cell carcinoma (HNSCC). In the anabolic pathway of 5-FU, the first step in activation of the drug is phosphorylation of 5-FU by orotate phosphoribosyltransferase (OPRT), which directly metabolizes 5-FU to 5-fluorouridine monophosphate (FUMP) in the presence of 5-phosphoribosyl-1-pyrophosphate. To date, OPRT expression in the tumors has been related to the clinical response or survival of cancer patients receiving 5-FU-based chemotherapy. In this study, we examined whether OPRT expression correlates with the chemosensitivity to 5-FU and cell proliferation in HNSCC. We constitutively expressed an OPRT cDNA in an HNSCC cell line. The effects of OPRT expression on in vitro cell growth and 5-FU cytotoxicity were examined. OPRT transfection increases the cytotoxicity of 5-FU without affecting cell proliferation of HNSCC cells in vitro. These results indicate that OPRT expression plays an important role in the sensitivity of HNSCC to 5-FU chemotherapy.


Is prolactin involved in sperm capacitation?
Immunoreactive prolactin in high concentration was detected in the human semen. Therefore, the effect of prolactin on the various metabolic parameters of the human spermatozoa was studied. It was found that prolactin brought about many of the biochemical changes in the spermatozoa which are known to be associated with the process of capacitation. These results suggest that prolactin has a role in the sperm capacitation.


Hepatitis in nursing homes. Incidence and management strategies.
Due to improved sanitation and prevention of parenterally transmitted diseases, a trend towards a decreasing incidence of acute viral hepatitis has been observed in several countries in the last years. These changes in epidemiology affect elderly people in different ways, especially if they are residents of homes for the aged. The decrease of hepatitis A virus (HAV) infection in childhood has resulted in less antibody protection during adulthood and old age. This in turn has led to an increased risk of acute and severe hepatitis due to HAV infection, and/or community outbreaks even in elderly people. The spread of parenterally transmitted viruses, such as hepatitis B virus (HBV), hepatitis C virus (HCV) and hepatitis delta virus (HDV), occurs in homes for the aged, mainly through the common use of toiletries and nondisposable syringes. Improvement in hygiene standards has lead to a marked decrease of these infections. In conclusion, provided that the current hygiene standards are maintained, the risk of parenterally transmitted hepatitis viruses (HBV, HCV and HDV) in homes for the aged will be low. Paradoxically, the risk of enterically transmitted HAV may, in the near future, increase in wealthy institutions and/or in Western countries.


Immediate postpartum assessment of the anal sphincter by endovaginal ultrasound: An experimental study.
Obstetric anal sphincter injuries are frequently missed and carry a significant risk for the development of anal incontinence. Immediate postpartum endoanal ultrasound increases identification of these injuries but is rarely employed. We hypothesize that endovaginal ultrasound could be a feasible and easily available alternative sonographic tool to improve early diagnosis of anal sphincter tears. We conducted a prospective experimental study including 160 primiparous women. Shortly after vaginal delivery, patients underwent clinical and sonographic perineal examinations. We analyzed the feasibility of anal sphincter assessment by endovaginal ultrasound and its potential contribution in the early diagnosis of anal sphincter injuries. Sonographic assessment of the anal sphincter was analyzable for 136 patients (85.0%). Causes of non-analyzability included air artifacts (6.9%), lack of distinction between the external anal sphincter and surrounding tissues (9.4%) and distortion artifacts (9.4%). Patients in the non-analyzable ultrasound subgroup were less likely to have delivered in a dorsal lithotomy position (62.5% vs. 85.3 %) and more likely to have had an episiotomy (33.3% vs. 14.0%), and their risk of sphincter injury was more frequently classified as "improbable" on clinical examination (91.7% vs. 61.0%). Ultrasounds were analyzable for 96.4% of patients clinically reported as having "possible" or "certain" sphincter injuries. The incidence of anal sphincter injury was 16.9% for clinical observation and 20.0% with associated sonographic examination. Endovaginal ultrasound could be used as a complementary tool in assessment of the anal sphincter in high-risk patients. Its feasibility and easy availability make this technique a promising tool for improving the management of anal sphincter tears.


Drug-induced immune hemolytic anemia associated with anti-vancomycin complicated by a paraben antibody.
Drug-induced immune hemolytic anemia (DIIHA) is rare, but potentially life-threatening. A high index of clinical suspicion is required for diagnosis, since the number of medications known to induce DIIHA continues to expand. Additionally, in vitro antibody reactivity against reagent additives has been reported, which may complicate test interpretation. A 61-year-old group A, D+ woman with a history of negative antibody detection tests developed hemolytic anemia on Postoperative Day 7 after repeat incision and drainage of a chronically infected right knee prosthesis. She was treated with multiple antibiotics in the postoperative period, including three cephalosporins and vancomycin intravenously as well as vancomycin and gentamicin-containing intraarticular cement spacers. A workup for possible DIIHA was performed. Testing was performed using vancomycin and cephalosporin antibiotics. Initially, gentamicin injection solution was used for testing, followed by testing with its component ingredients. A vancomycin antibody was detected and anemia resolved after vancomycin was discontinued. Reactivity was seen when gentamicin injection solution was used for testing, raising the possibility of a gentamicin antibody as well. However, testing with purified gentamicin as well as methylparaben and propylparaben demonstrated a paraben antibody that reacted with the paraben-containing gentamicin solution. The patient also demonstrated an anti-N. Neither the paraben antibody nor the anti-N appeared to cause in vivo hemolysis. This is the second reported case of DIIHA associated with anti-vancomycin. It is the fourth report describing a paraben antibody.


Mapping of the binding site for Mannheimia haemolytica leukotoxin within bovine CD18.
To map the site involved in Mannheimia haemolytica leukotoxin (LktA) binding and biological activity within bovine CD18, bovine x human CD18 chimeric constructs were generated and coexpressed with bovine CD11a in K562 cells. Studies with the chimeric leukocyte function-associated antigen 1 transductants demonstrate that the site required for LktA binding and biological effects resides within amino acid residues 500 and 600 of the extracellular region of bovine CD18.


Sterol and lipid metabolism in bees.
Bees provide essential pollination services for many food crops and are critical in supporting wild plant diversity. However, the dietary landscape of pollen food sources for social and solitary bees has changed because of agricultural intensification and habitat loss. For this reason, understanding the basic nutrient metabolism and meeting the nutritional needs of bees is becoming an urgent requirement for agriculture and conservation. We know that pollen is the principal source of dietary fat and sterols for pollinators, but a precise understanding of what the essential nutrients are and how much is needed is not yet clear. Sterols are key for producing the hormones that control development and may be present in cell membranes, where fatty-acid-containing species are important structural and signalling molecules (phospholipids) or to supply, store and distribute energy (glycerides). In this critical review, we examine the current general understanding of sterol and lipid metabolism of social and solitary bees from a variety of literature sources and discuss implications for bee health. We found that while eusocial bees are resilient to some dietary variation in sterol supply the scope for this is limited. The evidence of both de novo lipogenesis and a dietary need for particular fatty acids (FAs) shows that FA metabolism in insects is analogous to mammals but with distinct features. Bees rely on their dietary intake for essential sterols and lipids in a way that is dependent upon pollen availability.


The effects of n-acetylcysteine and/or deferoxamine on manic-like behavior and brain oxidative damage in mice submitted to the paradoxal sleep deprivation model of mania.
Bipolar disorder (BD) is a severe psychiatric disorder associated with social and functional impairment. Some studies have strongly suggested the involvement of oxidative stress in the pathophysiology of BD. Paradoxal sleep deprivation (PSD) in mice has been considered a good animal model of mania because it induces similar manic-like behavior, as well as producing the neurochemical alterations which have been observed in bipolar patients. Thus, the objective of the present study was to evaluate the effects of the antioxidant agent's n-acetylcysteine (Nac) and/or deferoxamine (DFX) on behavior and the oxidative stress parameters in the brains of mice submitted to the animal model of mania induced by PSD. The mice were treated for a period of seven days with saline solution (SAL), Nac, DFX or Nac plus DFX. The animals were subject to the PSD protocol for 36 h. Locomotor activity was then evaluated using the open-field test, and the oxidative stress parameters were subsequently evaluated in the hippocampus and frontal cortex of mice. The results showed PSD induced hyperactivity in mice, which is considered a manic-like behavior. In addition to this, PSD increased lipid peroxidation and oxidative damage to proteins, as well as causing alterations to antioxidant enzymes in the frontal cortex and hippocampus of mice. The Nac plus DFX adjunctive treatment prevented both the manic-like behavior and oxidative damage induced by PSD. Improving our understanding relating to oxidative damage in biomolecules, and the antioxidant mechanisms presented in the animal models of mania are important in helping to improve our knowledge concerning the pathophysiology and development of new therapeutical treatments for BD.


[Circumscribed intrapulmonary densities following thoracic trauma (author's transl)].
Intrapulmonary hematoma caused by blunt trauma to the chest may appear as round opacities of various size. The diagnosis depends on an exact case history and radiological follow up. 4 relevant cases are being presented. The incidence and pathogeneses are being discussed.


Comparative biomechanical analysis of gait in patients with central cord and Brown-Séquard syndrome.
This is a pilot study with the aim to highlight the use of kinematic and kinetic analyses as an adjunct to the assessment of individual patients with central cord syndrome (CCS) and hemisection or Brown-Séquard syndrome (BSS) and to discuss their possible consequences for clinical management. The sample studied consisted of 17 patients with CCS, 13 with BSS and 20 control subjects (control group (CG)). Data were obtained using a three-dimensional motion analysis system and two force plates. Gait differences were compared between CCS, BSS walking at a self-selected speed and CG at both a self-selected and a similar speed to that of the patient groups. The most relevant findings involved the knee and ankle, especially in the sagittal plane. In patients with CCS, knee flexion at initial contact was increased with respect to those in the BSS group (p < 0.01). The ankle in the BSS group made initial contact with a small degree of plantar flexion. The use of gait biomechanical analysis to detect underlying impairments can help the physician to set a specific rehabilitation program in each CCS and BSS walking patient. In this group of patients, rehabilitation treatment should aim to improve gait control and optimise ankle positioning at initial contact. Implications for Rehabilitation In this study, gait differences between patients with CSS and BSS were evaluated with biomechanical equipment. The most remarkable differences were found in the knee and ankle sagittal plane due to ankle position at initial contact. In this group of patients, rehabilitation treatment should aim to improve gait control and to get a better ankle positioning at initial contact.


Impact of participation in randomized trials on outcome following surgery for gastro-oesophageal reflux.
Patients may be unwilling to participate in clinical trials if they perceive risks. Outcomes were evaluated following surgery for gastro-oesophageal reflux in patients recruited to randomized trials compared with patients not in trials. This study compared outcomes of patients who had surgery for reflux within or outside randomized trials between 1994 and 2009. The choice of procedure outside each trial was according to surgeon or patient preference. Clinical outcomes were determined 1 and 5 years after surgery using a standardized questionnaire, with analogue scales to assess heartburn, dysphagia and overall satisfaction. Subgroup analysis was undertaken for those aged less than 75 years undergoing laparoscopic Nissen fundoplication. Some 417 patients entered six randomized trials evaluating surgery for reflux and 981 underwent surgery outside the trials. The trial group contained a higher proportion of men and younger patients, and patients in trials were more likely to have undergone Nissen fundoplication. At 1 year, patients in the trials had slightly lower heartburn scores and less abdominal bloating, but otherwise similar outcomes to those not in the trials. At 5 years there were no differences, except for a slightly higher dysphagia score for liquids in the trial group. For the subgroup analysis, demographic data were similar for both groups. There were no differences at 1 year, but at 5 years patients enrolled in the trials had higher scores for dysphagia for liquids and heartburn. All of the statistically significant differences were thought unlikely to be clinically relevant. Participation in a randomized trial assessing surgery for reflux did not influence outcomes.


Antioxidant properties of lipid concomitants in edible oils: A review.
Edible oils are indispensable for human life, providing energy and necessary fatty acids. Nevertheless, they are vulnerable to oxidation via a number of different mechanisms. Essential nutrients deteriorate as well as toxic substances are produced when edible oils are oxidized; thus, they should be retarded wherever possible. Lipid concomitants have a strong antioxidant capacity and are a large class of biologically active chemical substances in edible oils. They have shown remarkable antioxidant properties and were documented to improve the quality of edible oils in varied ways. An overview of the antioxidant properties of the polar, non-polar, and amphiphilic lipid concomitants present in edible oils is provided in this review. Interactions among various lipid concomitants and the probable mechanisms are also elucidated. This review may provide a theoretical basis and practical reference for food industry practitioners and researchers to understand the underlying cause of variations in the quality of edible oils.


Establishment of reference ranges for ductus venosus waveform indices in the Japanese population.
To establish reference ranges for ductus venosus waveform indices in the Japanese population. In this retrospective cross-sectional study, 791 singleton fetuses of healthy Japanese couples were examined from January 2004 to January 2008. Reference ranges for ductus venosus waveform indices were constructed from cross-sectional data obtained at between 18 and 41 weeks of gestation. With a success rate of 84%, a total of 667 measurements in 791 women were eligible for analysis. The median pulsatility index (PI) of fetal ductus venosus decreased from 0.54 at 18 weeks of gestation to 0.30 at 41 weeks of gestation. The median end-diastolic velocity/peak systolic velocity (a/S) of the ductus venosus increased from 0.56 at 18 weeks of gestation to 0.76 at 41 weeks of gestation. In this study, we established reference ranges for the PI and a/S of the ductus venosus in the Japanese population, which differed slightly from other published reference data. The results will be useful for further studies to determine the validity of the clinical importance of the ductus venosus for at-risk fetuses.


Can the green finance improve export green-sophistication? New insights from China.
The green financial policy is one of the important policy tools for China to achieve the national carbon peak goal and carbon neutrality through financial means. How financial development affects the growth of international trade has been an important research topic. This paper uses the Pilot Zones for Green Finance Reform and Innovations (PZGFRI) implemented in 2017 as a natural experiment drawing on the relevant data of Chinese provinces' panel data from 2010 to 2019. It adopts a difference in difference (DID) model to assess the impact of green finance on export green-sophistication (EGS). The results report that the PZGFRI significantly improves EGS, and the result remains robust after robustness checks such as parallel trend and placebo. The PZGFRI improves EGS by boosting total factor productivity, industrial structure upgrading, and green technology innovation. Moreover, the role of PZGFRI in promoting EGS is significantly reflected in the central and western regions and the regions with low-marketization levels. This study confirms that green finance is an important factor influencing the quality improvement of China's exports, which provides effective evidence from the reality level for China to vigorously promote the construction of a green financial system in recent years.


A Fourier transform infrared spectroscopic study of the molecular interaction of cholesterol with 1,2-dipalmitoyl-sn-glycero-3-phosphocholine.
The temperature dependencies of the infrared spectra of pure and cholesterol-containing multibilayers of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine were studied using Fourier transform infrared techniques. A comparison of the spectroscopic data showed the retention of a melting phenomenon at 60 mol% cholesterol content, and the retention of some all-trans conformations in the liquid-crystalline phase. It is also demonstrated that at temperatures less than 30 degrees C, the cholesterol-containing 1,2-dipalmitoyl-sn-glycero-3-phosphocholine multibilayers still contain a small amount of pure 1,2-dipalmitoyl-sn-glycero-3-phosphocholine, packed in an orthorhombic subcell lattice. Spectral changes were found in the absorptions characteristic of the phospholipid head groups. The addition of cholesterol results in changes in the ester bands, and demonstrates the induction by cholesterol of non-equivalent ester conformations.


Analysis of several methods for the extraction of high quality DNA from acetic acid bacteria in wine and vinegar for characterization by PCR-based methods.
Acetic acid bacteria (AAB) are fastidious microorganisms with poor recovery in culture. Culture-independent methods are currently under examination. Good DNA extraction is a strict requirement of these methods. We compared five methods for extracting the DNA of AAB directly from wine and vinegar samples. Four matrices (white wine, red wine, superficial vinegar and submerged vinegar) contaminated with two AAB strains belonging to Acetobacter pasteurianus and Gluconacetobacter hansenii were assayed. To improve the yield and quality of the extracted DNA, a sample treatment (washing with polyvinyl pyrrolidone or NaCl) was also tested. DNA quality was measured by amplification of the 16S rRNA gene with conventional PCR. DNA recovery rate was assessed by real-time PCR. DNA amplification was always successful with the Wizard method though DNA recovery was poor. A CTAB-based method and NucleoSpin protocol extracted the highest DNA recoveries from wine and vinegar samples. Both of these methods require treatment to recover suitable DNA for amplification with maximum recovery. Both may therefore be good solutions for DNA extraction in wine and vinegar samples. DNA extraction of Ga hansenii was more effective than that of A. pasteurianus. The fastest and cheapest method we evaluated (the Thermal shock protocol) produced the worst results both for DNA amplification and DNA recovery.


Reference values of osteocalcin and procollagen type I N-propeptide plasma levels in a healthy Central European population aged 0-18 years.
The aim of this study was to assess the relationships between both a marker of bone formation and a marker of bone turnover and age, sex, and pubertal stage in a group (n = 439) of healthy children and adolescents. These reference data should be instrumental in interpretation of results. The skeletal system has high metabolic activity. In children, bone markers may be useful in diagnostics and treatment management of skeletal diseases but there could be difficulties with interpretation of results. Compared with adults, children have elevated bone marker levels due to high skeletal growth velocity and rapid bone turnover. Thus, valid age- and sex-specific reference data should be obtained for each pediatric population living in a particular climate and with a similar lifestyle. The aim of this study was to assess the relationships between both a marker of bone formation (procollagen type I N-terminal propeptide [PINP]) and a marker of bone turnover (osteocalcin [OC]) and age, sex, and pubertal stage in a group of healthy children and adolescents. Four hundred thirty-nine healthy Caucasian children participated. Their height, weight, and pubertal stage were recorded. Fasting PINP and OC were measured using a morning blood sample. The highest levels of PINP were observed during the first year of life. There is no OC postnatal peak, but levels are higher than the adult reference interval throughout childhood. OC peaks with the pubertal growth spurt at second-third Tanner stage of breast development in girls and at second-third Tanner stage of genital development in boys. PINP peaks during second-third Tanner stage of breast development in girls and at third Tanner stage of genital development in boys. This study provides reference data for OC and PINP in healthy Caucasian children from a Central European population.


Pubertal maturation and time of day differentially affect behavioral and neuroendocrine responses following an acute stressor.
Puberty markedly influences stress responsiveness such that prepubertal animals show a more protracted corticosterone (CORT) and progesterone response following acute stress compared to adults. In both adult and juvenile rats, circadian time modulates adrenocortical steroids with basal CORT and progesterone levels rising prior to the onset of the dark phase of the light-dark cycle (i.e., active period). How time of day affects the pubertal difference in stress responsiveness and if the behaviors of prepubertal and adult animals are differentially affected by stress and time of testing remain unknown. Thus, we exposed group housed (3 per cage) prepubertal (28d) and adult (77d) male rats to 30 min of restraint in either the early portion of the behaviorally inactive, light (circadian nadir of CORT and progesterone) or behaviorally active, dark (circadian peak) phase of their light-dark cycle and measured ACTH, CORT, progesterone, and home cage behavior before and after the stressor. We found that the extended hormonal stress response demonstrated by prepubertal males occurred at both times of day. However, differences in post-stress behavior were dependent on time of testing. Specifically, although pre- and post-stress behaviors were similarly affected by the stressor in the light phase in prepubertal and adult males, during the dark phase, stress suppressed play behavior in the prepubertal males, and increased their time spent resting together (huddling), while these behaviors were unaffected by stress in the adults. These data indicate that pubertal development and time of day interact to modulate post-stress behavior and demonstrate a dissociation between post-stress hormonal and behavioral responses.


Live monitoring of small vessels during development and disease using the flt-1 promoter element.
Vessel formation is of critical importance for organ function in the normal and diseased state. In particular, the labeling and quantitation of small vessels prove to be technically challenging using current approaches. We have, therefore, established a transgenic embryonic stem (ES) cell line and a transgenic mouse model where the vascular endothelial growth factor receptor VEGFR-1 (flt-1) promoter drives the expression of the live reporter eGFP. Fluorescence microscopy and immunostainings revealed endothelial-specific eGFP labeling of vascular networks. The expression pattern recapitulates that of the endogenous flt-1 gene, because small and large vessels are labeled by eGFP during embryonic development; after birth, the expression becomes more restricted to small vessels. We have explored this in the cardiovascular system more in detail and found that all small vessels and capillaries within the heart are strongly eGFP+. In addition, myocardial injuries have been induced in transgenic mice and prominent vascular remodeling, and an increase in endothelial cell area within the peri-infarct area could be observed underscoring the utility of this mouse model. Thus, the transgenic flt-1/eGFP models are powerful tools to investigate and quantify vascularization in vivo and to probe the effect of different compounds on vessel formation in vitro.


Reassessing the Use of Proton Pump Inhibitors and Histamine-2 Antagonists in Critically Ill Children: A Systematic Review and Meta-Analysis.
To determine the associations of stress ulcer prophylaxis with gastrointestinal (GI) bleeding, nosocomial pneumonia (NP), mortality, and length of stay in the pediatric intensive care unit (PICU). We conducted a systematic review and meta-analysis of randomized controlled trials (RCTs) and observational studies in the English language assessing the effects of proton pump inhibitors and histamine-2 receptor antagonists on patients in the PICU published before October 2018 from the PubMed, Embase, CINAHL, and Cochrane Central Register of Controlled Trials databases. A random-effects Mantel-Haenszel risk difference (MHRD) model was used to pool all the selected studies for meta-analysis. Primary outcomes were the incidences of GI bleeding and NP. Secondary outcomes included mortality and length of PICU stay. Seventeen studies (4 RCTs and 13 observational studies) with a total of 340 763 patients were included. The overall incidence of GI bleeding was 15.2%. There was no difference in the risk of GI bleeding based on stress ulcer prophylaxis status (MHRD, 5.0%; 95% CI, -1.0% to 11.0%; I2 = 62%). There was an increased risk of NP in patients who received stress ulcer prophylaxis compared with those who did not (MHRD, 5.3%; 95% CI, 3.5%-7.0%; I2 = 0%). An increased risk of mortality was seen in patients receiving stress ulcer prophylaxis (MHRD, 2.1%; 95% CI, 2.0%-2.2%; I2 = 0%), although this association was no longer found when 1 large study was removed in a sensitivity analysis. There was no statistically significant difference in length of PICU stay between the groups (standardized mean difference, 0.42 days; 95% CI, -0.16 to 1.01 days; I2 = 89.8%). Stress ulcer prophylaxis does not show a clear benefit in reducing GI bleeding or length of PICU stay. Observational studies suggest an increased risk of NP and mortality with stress ulcer prophylaxis, which remains to be validated in clinical trials.


Intrinsic layer based automatic specular reflection detection in endoscopic images.
Endoscopic images are used to observe the internal structure of the human body. Specular reflection (SR) images are mostly a consequence of the strong light and bright regions appearing on endoscopic images, which affects the performance of minimally invasive surgery. In this study, we propose a novel method for automatic SR detection based on intrinsic image layer separation (IILS). The proposed method consists of three steps. Initially, it involves the normalization of the image followed by the extraction of high gradient area, and the separation of SR is done on the basis of the color model. The image melding technique is utilized to reconstruct the reflected pixels. The experiments were conducted on 912 endoscopic images from CVC-EndoSceneStill. The results of accuracy, sensitivity, specificity, precision, Jaccard index, Dice coefficient, standard deviation, and pixel count difference show that the detection performance of the proposed method outperforms that of other state-of-the-art methods. The evaluation of the proposed IILS-based SR detection demonstrates that our method obtains better qualitative and quantitative assessments compared with other methods, which can be used as a promising preprocessing step for further analysis of endoscopic images.


Biodegradation and kinetics of aerobic granules under high organic loading rates in sequencing batch reactor.
Biodegradation, kinetics, and microbial diversity of aerobic granules were investigated under a high range of organic loading rate 6.0 to 12.0 kg chemical oxygen demand (COD) m(-3) day(-1) in a sequencing batch reactor. The selection and enriching of different bacterial species under different organic loading rates had an important effect on the characteristics and performance of the mature aerobic granules and caused the difference on granular biodegradation and kinetic behaviors. Good granular characteristics and performance were presented at steady state under various organic loading rates. Larger and denser aerobic granules were developed and stabilized at relatively higher organic loading rates with decreased bioactivity in terms of specific oxygen utilization rate and specific growth rate (muoverall) or solid retention time. The decrease of bioactivity was helpful to maintain granule stability under high organic loading rates and improve reactor operation. The corresponding biokinetic coefficients of endogenous decay rate (kd), observed yield (Yobs), and theoretical yield (Y) were measured and calculated in this study. As the increase of organic loading rate, a decreased net sludge production (Yobs) is associated with an increased solid retention time, while kd and Y changed insignificantly and can be regarded as constants under different organic loading rates.


Therapeutic oral sesame oil is ineffectual against monocrotaline-induced sinusoidal obstruction syndrome in rats.
The chemotherapeutic drug oxaliplatin causes sinusoidal obstruction syndrome (SOS), which is analogous to that of monocrotaline-induced SOS. Sesame oil is a nutrient-rich antioxidant in alternative medicine. It contains phenol, sesamin, sesamol, and sesamolin, all of which contribute to its antioxidant property. The authors investigated the therapeutic effect of oral sesame oil against monocrotaline-induced SOS in rats. Male Sprague-Dawley rats were gavaged with monocrotaline (90 mg/kg) to induce SOS. Control rats were treated with saline only at 0 and 24 hours. Sesame oil (0.5, 1, 2, or 4 mL/kg, orally) was given 24 hours after monocrotaline in rats. Blood samples were collected at 24 and 48 hours after monocrotaline was given to assess the level of aspartate aminotransferase (AST) and alanine aminotransferase (ALT). Histopathology was also assessed 48 hours after monocrotaline was given. AST and ALT were significantly higher in monocrotaline-treated rats than in control rats. Oral sesame oil did not decrease AST and ALT in monocrotaline-treated rats. In addition, liver pathology revealed that oral sesame oil had no therapeutic effect against SOS. Oral sesame oil is therapeutically ineffectual against monocrotaline-induced SOS.


Use of a modified binocular visual field to assess cyclodiplopia.
The patient who notes binocular diplopia may indicate that horizontal, vertical, or cyclorotary components are present. Although the single binocular visual field provides an indication of the area which is free of horizontal and vertical diplopia, it does not assess cyclorotary diplopia. A simple modification of the single binocular visual field which allows determination of the clinical significance of the cyclorotary component of diplopia is presented. Reproducible information is easily obtained, assisting in patient follow-up and management.


Spatial variations in the incidence of breast cancer and potential risks associated with soil dioxin contamination in Midland, Saginaw, and Bay Counties, Michigan, USA.
High levels of dioxins in soil and higher-than-average body burdens of dioxins in local residents have been found in the city of Midland and the Tittabawassee River floodplain in Michigan. The objective of this study is threefold: (1) to evaluate dioxin levels in soils; (2) to evaluate the spatial variations in breast cancer incidence in Midland, Saginaw, and Bay Counties in Michigan; (3) to evaluate whether breast cancer rates are spatially associated with the dioxin contamination areas. We acquired 532 published soil dioxin data samples collected from 1995 to 2003 and data pertaining to female breast cancer cases (n = 4,604) at ZIP code level in Midland, Saginaw, and Bay Counties for years 1985 through 2002. Descriptive statistics and self-organizing map algorithm were used to evaluate dioxin levels in soils. Geographic information systems techniques, the Kulldorff's spatial and space-time scan statistics, and genetic algorithms were used to explore the variation in the incidence of breast cancer in space and space-time. Odds ratio and their corresponding 95% confidence intervals, with adjustment for age, were used to investigate a spatial association between breast cancer incidence and soil dioxin contamination. High levels of dioxin in soils were observed in the city of Midland and the Tittabawassee River 100-year floodplain. After adjusting for age, we observed high breast cancer incidence rates and detected the presence of spatial clusters in the city of Midland, the confluence area of the Tittabawassee, and Saginaw Rivers. After accounting for spatiotemporal variations, we observed a spatial cluster of breast cancer incidence in Midland between 1985 and 1993. The odds ratio further suggests a statistically significant (alpha = 0.05) increased breast cancer rate as women get older, and a higher disease burden in Midland and the surrounding areas in close proximity to the dioxin contaminated areas. These findings suggest that increased breast cancer incidences are spatially associated with soil dioxin contamination. Aging is a substantial factor in the development of breast cancer. Findings can be used for heightened surveillance and education, as well as formulating new study hypotheses for further research.


[Comparative evaluation of the use of nalbuphine and buprenorphine in prehospital care].
Analgesic efficacy of nalbufine and buprenorphine is assessed in 86 patients with painful syndrome caused by unstable angina and acute myocardial infarction and in 72 patients with locomotor injuries, to whom urgent care was rendered by ambulance teams before hospitalization. By the velocity and depth of analgesic effect nalbufine is not inferior to morphine, and in patients with unstable angina and myocardial infarction is even superior to it. High analgesic activity of buprenorphine is compatible to that of morphine, but the rate of analgesia development is insufficient for urgent care.


Irritability uniquely predicts prefrontal cortex activation during preschool inhibitory control among all temperament domains: A LASSO approach.
Temperament, defined as individual variation in the reactivity and regulation of emotional, motor, and attentional processes, has been shown to influence emotional and cognitive development during the preschool period (ages 4-5). While relationships between temperament and neural activity have been investigated previously, these have typically investigated individual temperament dimensions selected ad hoc. Since significant correlations exist between various temperament dimensions, it remains unclear whether these findings would replicate while analyzing all temperament dimensions simultaneously. Using functional near infrared spectroscopy (fNIRS), 4-5-year-old children (N = 118) were administered a Go/No-Go task to assess prefrontal cortex activation during inhibitory control. The relationship between PFC activation and all 15 temperament domains defined by the Children's Behavior Questionnaire (CBQ) was assessed using automatic feature selection via LASSO regression. Results indicate that only the Anger/Frustration dimension was predictive of activation during the inhibitory control task. These findings support previous work showing relationships between irritability and prefrontal activation during executive function and extend those findings by demonstrating the specificity of the activation-irritability relationship among temperament dimensions.


Oligomerization, trans-reduction, and instability of mutant NOTCH3 in inherited vascular dementia.
Cerebral small vessel disease (SVD) is a prevalent disease of aging and a major contributor to stroke and dementia. The most commonly inherited SVD, CADASIL, is caused by dominantly acting cysteine-altering mutations in NOTCH3. These mutations change the number of cysteines from an even to an odd number, but the impact of these alterations on NOTCH3 protein structure remain unclear. Here, we prepared wildtype and four mutant recombinant NOTCH3 protein fragments to analyze the impact of CADASIL mutations on oligomerization, thiol status, and protein stability. Using gel electrophoresis, tandem MS/MS, and collision-induced unfolding, we find that NOTCH3 mutant proteins feature increased amounts of inappropriate disulfide bridges, reduced cysteines, and structural instability. Presence of a second protein factor, an N-terminal fragment of NOTCH3 (NTF), is capable of further altering disulfide statuses of both wildtype and mutant proteins, leading to increased numbers of reduced cysteines and further destabilization of NOTCH3 structure. In sum, these studies identify specific cysteine residues alterations and quaternary structure induced by CADASIL mutations in NOTCH3; further, we validate that reductive factors alter the structure and stability of this small vessel disease protein.


Comparative hemodynamic effects of OPC-18790 and dobutamine in patients with advanced heart failure.
OPC-18790 (Otsuka America Pharmaceutical, Rockville, MD), a novel positive inotropic agent, produces titratable hemodynamic benefits in patients with advanced heart failure. In such patients, OPC-18790 has been shown to acutely increase the cardiac index, while reducing systemic vascular resistance and left ventricular filling pressure, without an associated increase in heart rate. This study was performed to compare the acute hemodynamic effects of OPC-18790 and the beta-adrenergic receptor agonist, dobutamine, in patients with advanced heart failure. OPC-18790 and dobutamine were compared on successive days in 13 patients with worsening New York Heart Association class III or IV heart failure. The mean (+/- SEM) left ventricular ejection fraction was 15 +/- 2% (range, 6-29%). Pretreatment hemodynamics were: heart rate, 96 +/- 2 beats/min; mean arterial pressure, 77 +/- 3 mmHg; cardiac index, 1.80 +/- 0.10 L/min/m2; pulmonary capillary wedge pressure, 27 +/- 1 mmHg; mean pulmonary arterial pressure, 41 +/- 2 mmHg; and systemic vascular resistance, 1,732 +/- 152 dynes.s/cm5. At infusion rates yielding comparable increases in the cardiac index (5 micrograms/kg/min for 2 hours for each drug), OPC-18790 produced significantly more favorable effects on heart rate (-2 +/- 3% vs 11 +/- 4%; P = .01), pulmonary capillary wedge pressure (-32 +/- 4% vs -17 +/- 8%; P = .04), mean pulmonary arterial pressure (-14 +/- 3% vs 6 +/- 11%; P = .06), stroke volume index (48 +/- 8% vs 29 +/- 7%; P = .02), stroke work index (70 +/- 11 vs 42% +/- 12%; P = .03), and rate pressure product (2 +/- 4% vs 14 +/- 4%; P = .05). The hemodynamic profile for OPC-18790 differs from dobutamine, with OPC-18790 exhibiting no increase in heart rate, greater preload reduction, and an increase in cardiac performance at a lower estimated metabolic cost.


Folding alpha-helical membrane proteins into liposomes in vitro and determination of secondary structure.
The native environment of integral membrane proteins is a highly complex lipid bilayer composed of many different types of lipids, the physical characteristics of which can profoundly influence protein stability, folding, and function. Secondary transporters are a class of protein where changes to both structure and activity have been observed in different bilayer environments. In order to study these interactions in vitro, it is necessary to extract and purify the protein and exchange it into an artificial lipid system that can be manipulated to control protein behavior. Liposomes are a commonly used model system that is particularly suitable for studying transporters. GalP and LacY can be reconstituted or refolded into vesicles with a high degree of efficiency for further structural analysis. Circular dichroism spectroscopy is an important technique in monitoring protein folding, which allows the decomposition of spectra into secondary structural components.


Effect of erector spinae plane block with different doses of dexmedetomidine as adjuvant for ropivacaine on the postoperative quality of recovery after video-assisted thoracoscopic lobectomy surgery: a randomized controlled trial.
Erector spinae plane block (ESPB) improves postoperative analgesia and significantly enhances the quality of recovery (QoR) after video-assisted thoracoscopic lobectomy surgery (VATLS). However, it is not known whether the use of dexmedetomidine (Dex) as an adjunct for ropivacaine to ESPB affects the QoR after VATLS. The purpose of this study was to explore the effects of different Dex dosages as an adjunct for ropivacaine in combination with ultrasound-guided ESPB on the quality of postoperative recovery in patients with VATLS. In this single-center, double-blind, randomized study, 120 patients between the ages of 18 and 65 who were scheduled for VATLS from december 2021 and october 2022 in our hospital under general anesthesia were randomly divided into three groups: ultrasound-guided ESPB with 30 mL of 0.5% ropivacaine (Group R), ultrasound-guided ESPB 0.5% ropivacaine plus 0.5 µg/kg Dex (Group RD1), and ultrasound-guided ESPB 0.5% ropivacaine plus 1.0 µg/kg Dex (Group RD2), ultrasound-guided ESPB was administrated at the T5 vertebral level before surgery. The primary outcome was the QoR-15 score 24 h after the surgery. The secondary outcomes included the QoR-15 scores at 12 h, 48 h, and 72 h after the operation, visual analogue scale (VAS) scores at 8 h, 12 h, 24 h, and 48 h after surgery, cumulative flurbiprofen consumption, postoperative nausea and vomiting (PONV), postoperative bradycardia, and hypotension. The QoR-15 scores were higher in group RD2 than the R and RD1 groups on postoperative day 1 (P < 0.05), in addition, no significant difference was found in the QoR-15 scores between groups R and RD1 on postoperative day 1. The VAS scores were significantly lower in group RD2 than in groups RD1 and group R 12-24 h after surgery (P < 0.05). No significant differences were observed in the QoR-15 and VAS scores at 48 and 72 h after surgery between the three groups. The cumulative flurbiprofen consumption was markedly reduced during the 72 h after surgery in the RD2 group (P < 0.05). The incidence of postoperative nausea and vomiting was lower in the RD2 group (P < 0.05). The combination of 1 µg/kg dexmedetomidine as an adjunct with 0.5% ropivacaine 30 ml for erector spinae plane block significantly improved the postoperative quality of recovery and provided better postoperative analgesia on postoperative day 1 in patients undergoing Video-assisted thoracoscopic lobectomy surgery. However, dexmedetomidine (1 µg/kg) as an adjunct for ropivacaine combined with erector spinae plane block did not enhance the postoperative quality of recovery at 48 and 72 h postoperatively. The number of this clinical trial registry is ChiCTR2100053230, date of registration: 16/11/ 2021).


Health management technology for catastrophic medical conditions.
The excessive growth of health care expenditures in the United States is widely acknowledged. Costs are anticipated to double by the year 2006. The intractable issue which remains before health care leaders is how to appropriately restrain these costs while not sacrificing a desired level of care quality. A variety of management approaches have been developed in pursuit of more rational and cost-effective use of health resources. Current management approaches have proven inadequate in stemming health care cost inflation and have raised increasing concerns about their negative impact on the quality of health care. One group has created and operated a data and structured, expert consensus-driven health management technology for the management of catastrophic medical conditions, including severe brain and spinal cord injury and severe multiple trauma and burns, since 1992 and has recently applied this same technology to high risk neonates and organ transplants. This integrated, severity risk adjusted, delivery system incorporates adequate clinical data capture and analysis, coupled with empirically derived management principles and consensus expert clinical judgment. Preliminary data analysis indicates that patients treated under the ParadigmHealth model experienced an improvement in the health care process, improved quality in health care delivery and outcomes, and overall cost reduction.


Circulating Tumour DNA Is a Strong Predictor of Outcomes in Patients Treated with Systemic Therapy for Urothelial Carcinoma.
We summarise the available data for and assess the prognostic value of circulating tumour DNA (ctDNA) in patients treated with systemic therapy for urothelial carcinoma (UC). Studies were deemed eligible if they reported on oncologic outcomes for patients with UC treated with systemic therapy according to the baseline ctDNA profile (before starting systemic therapy) and/or changes over the course of therapy. Five studies met the eligibility criteria. We found a strong association between high baseline ctDNA levels and worse disease-free survival (DFS; hazard ratio [HR] 3.53, 95% confidence interval [CI] 2.58-4.84) and overall survival (OS; HR 2.99, 95% CI 2.17-4.13). Patients with a decline in ctDNA level after immunotherapy had better DFS (HR 0.25, 95% CI 0.13-0.49) and OS (HR 0.10, 95% CI 0.03-0.42) in comparison to patients without a ctDNA decline. Conversely, an increase in ctDNA levels after immunotherapy was associated with worse survival outcomes. Patients with UC who exhibited a decrease in ctDNA levels during systemic therapy had better survival outcomes compared to those with stable or increasing ctDNA levels. PATIENT SUMMARY: Measurement of tumour DNA in blood may help in identifying patients with cancer of the urinary tract who are unlikely to respond to chemotherapy or immunotherapy. This could serve as a biomarker for monitoring cancer treatment.


[Survey of 319 women satisfaction using abortion in centers of Pays de la Loire in France].
Perinatal network the "Security birth" Pays de la Loire (RSN), in collaboration with the regional network "Sexual health" (RRSS) organized a satisfaction survey in 2014 among women who had an induced abortion in the centers in Pays de la Loire. The objective of the study is to evaluate the satisfaction of women who resort to abortion centers, study the factors and propose actions for improvement. A questionnaire, developed with professionals, was offered to women after the end of the abortion act. The scales of satisfaction were based on Likert scales to 4 degrees. They were collected for the period of contact centers, for the abortion itself, and overall. Data were collected on the civil status, the care pathway to get to the abortion center, features centers and professionals met, acts anesthesia and end of abortion, wait times and overall satisfaction of women. The women with rather poor satisfaction were compared with other women by univariate analysis with odds ratios (OR) and multivariate with adjusted odds ratios (ORa). Of the 18 centers, 13 participated and 319 responses were analyzed. The abortions were performed on average 8±SA 2. Among the methods, 40.1 % were drug, and 59.9 % with anesthesia (14.0 % with general anesthesia [AG] and 45.9 % under local anesthesia [AL]). The first contacts on average to 5.7 SA were mostly general practitioners (38 %), then the abortion centers directly (22 %), and planning centers (16 %). The information was considered clear (94 %), the appropriate orientation (97 %), with respect to the application (98 %). The first meeting at the center was done 7 days after the call (6.7 SA), and met women overall. The abortion was performed 16 days after the first call center (8.0 SA) with satisfaction by field 78 % (waiting time) to 98 % (confidentiality, privacy…). The overall satisfaction rate was 89.0 % and 8.2 % were not satisfied. Factors related to the non-satisfaction in multivariate analysis were the least easy access to the center (ORa=0.31 [.11 to 0.86; p=0.02]), pain perceived≥4 (ORa=3 50 [1.32 to 9.28], p=0.02), the lack of explanation (ORa=0.23 [0.05 to 0.97]; p=0.04), and the accompanying inability (ORa=0.27 [0.10 to 0.70], p=0.007). Finally, 12.6 % of women reported for improvement with clear remarks. This is the first regional survey on the satisfaction of women resorting to abortion. Delays from the first contact are satisfactory and show no organizational problem. The various dimensions of satisfaction showed a high satisfaction rate in the 13 participating centers. The least satisfactory factors are related to organizational problems (waiting circuits especially trips to the operating room and maternity) and problems related to the care itself (not enough explanations, reduced ability to reassure and support Savory absence, lack of choice of the method of anesthesia, pain experienced). Five areas for improvement have been proposed to the care-givers of the centers.


Trans-acting aptazyme for conditional gene knockdown in eukaryotic cells.
Trans-acting hammerhead ribozyme inherits the advantages of being the smallest and best-characterized RNA-cleaving ribozyme, offering high modularity and the ability to cleave any desired sequence without the aid of any protein, as long as the target sequence contains a cleavage site. However, achieving precise control over the trans-acting hammerhead ribozyme would enable safer and more accurate regulation of gene expression. Herein, we described an intracellular selection of hammerhead aptazyme that contains a theophylline aptamer on stem II based on toxin protein IbsC. Based on the intracellular selection, we obtained three new cis-acting hammerhead aptazymes. Moreover, the corresponding trans-acting aptazymes could be efficiently induced by theophylline to knock down different targeted genes in eukaryotic cells. Notably, the best one, T195, exhibited a ligand-dependent and dose-dependent response to theophylline, and the cleavage efficiency could be enhanced by incorporating multiplex aptazymes.


New strategies to identify protease substrates.
Proteome dynamics is governed by transcription, translation, and post-translational modifications. Limited proteolysis is an irreversible post-translational modification that generates multiple but unique proteoforms from almost every native protein. Elucidating these proteoforms and understanding their dynamics at a system-wide level is of utmost importance because uncontrolled proteolytic cleavages correlate with many pathologies. Mass spectrometry-based degradomics has revolutionized protease research and invented workflows for global identification of protease substrates with resolution down to precise cleavage sites. In this review, we provide an overview of current strategies in protease substrate degradomics and introduce the concept of workflow, mass spectrometry-based and in silico enrichment of protein termini with the perspective of full deconvolution of digital proteome maps for precision medicine, and degradomics biomarker diagnostics.


PRIMPOL-Mediated Adaptive Response Suppresses Replication Fork Reversal in BRCA-Deficient Cells.
Acute treatment with replication-stalling chemotherapeutics causes reversal of replication forks. BRCA proteins protect reversed forks from nucleolytic degradation, and their loss leads to chemosensitivity. Here, we show that fork degradation is no longer detectable in BRCA1-deficient cancer cells exposed to multiple cisplatin doses, mimicking a clinical treatment regimen. This effect depends on increased expression and chromatin loading of PRIMPOL and is regulated by ATR activity. Electron microscopy and single-molecule DNA fiber analyses reveal that PRIMPOL rescues fork degradation by reinitiating DNA synthesis past DNA lesions. PRIMPOL repriming leads to accumulation of ssDNA gaps while suppressing fork reversal. We propose that cells adapt to repeated cisplatin doses by activating PRIMPOL repriming under conditions that would otherwise promote pathological reversed fork degradation. This effect is generalizable to other conditions of impaired fork reversal (e.g., SMARCAL1 loss or PARP inhibition) and suggests a new strategy to modulate cisplatin chemosensitivity by targeting the PRIMPOL pathway.


Copper status and adriamycin treatment: effects on antioxidant status in mice.
The biochemical response of Cu-sufficient and Cu-deficient mice to adriamycin (ADR) treatment was evaluated. Mice were injected intraperitoneally with ADR (17 mg/kg body wt.) or saline (0.9% w/v) and killed 4 d after injection. There was no effect of ADR on cardiac superoxide dismutase (SOD) activity in Cu-sufficient of Cu-deficient mice. ADR injection resulted in higher cardiac glutathione (GSH) concentrations in Cu-sufficient mice while it resulted in lower GSH concentrations in Cu-deficient mice relative to their saline-injected controls. The effects of ADR in Cu-deficient mice were tissue-specific as its administration resulted in lower hepatic SOD activity and higher hepatic GSH concentrations relative to saline-injected controls.


Combined Effects of Diabetes and Education on Decline of Cognitive Performance in the Older Population: The Survey of Health, Ageing, and Retirement in Europe.
Diabetes management requires cognitive abilities. Unfortunately, diabetes is a risk factor for cognitive decline. However, the effects of diabetes in persons with reduced cognitive reserves are unclear. We aimed to examine whether the effects of diabetes and education on cognitive skills simply add up or deviate from simple additivity. We used data from waves 1 to 7 (except wave 3) of the Survey of Health, Ageing, and Retirement in Europe (SHARE) collected between 2004 and 2017. SHARE includes 140,000 persons aged ≥50 years from 28 European countries and Israel. Diabetes was assessed by self-report and education was coded according to the International Standard Classification of Education levels. Verbal fluency, immediate, and delayed memory were assessed using standard tests. Interaction between diabetes and education was estimated from interaction contrasts and from interaction terms in adjusted regression models. At baseline, cognitive performance declined in the order "no diabetes/high education" > "diabetes/high education" > "no diabetes/low education" > "diabetes/low education" for all three tests - e.g., the first group named 21.4 ± 7.2, the fourth group 14.6 ± 6.1 animals in a minute in the test of verbal fluency. Interaction contrasts and regression coefficients of interaction terms were close to zero, showing that the effects of diabetes and education on cognitive performance added up without interaction both in cross-sectional and longitudinal analyses. We have observed no interaction between education and diabetes on cognitive skills. Yet, people with diabetes and low education showed poor cognitive performance and should receive particular support in managing diabetes.


Gender and age influence the effects of slow-wave sleep on respiration in patients with obstructive sleep apnea.
Obstructive sleep apnea (OSA) is influenced by sleep architecture with rapid eye movement (REM) sleep having the most adverse influence, especially in women. There is little data defining the influence of slow-wave sleep (SWS) on OSA. We wished to study the influence of SWS on OSA and identify differences attributable to gender and/or age, if any. Retrospective study of polysomnography (PSG) records of adult patients referred for diagnostic PSG. Records were excluded if they underwent split night or positive airway pressure titration studies, had <180 min of total sleep time (TST) and/or <40% sleep efficiency, or had SWS <5 min and/or <1% of TST. The apnea-hypopnea index (AHI) recorded during SWS was compared with that measured during other non-rapid eye movement (NREM) sleep and during REM sleep. The REM-SWS difference in AHI was measured, and compared between genders. Records from 239 patients were included. The mean AHI in all subjects was 17.7 ± 22.6. The SWS AHI was 6.8 ± 18.9, compared to the REM AHI of 24.9 ± 25.8, and NREM AHI of 15.8 ± 22.8. Females had significantly higher SWS by percentage, and lower NREM AHI (P < 0.0001) and SWS AHI (P = 0.03). Among patients with OSA (AHI ≥5), the difference between REM AHI and SWS AHI was greater in women than in men (34.2 ± 27.4 vs. 21.6 ± 26.0, P = 0.006). The upper airway appears to be less susceptible to OSA during SWS than during REM and other NREM sleep. This may be related to phase-specific influences on both dynamic upper airway control as well as loop gain. Gender and age appear to modify this effect.


Effects of charge and lipophilicity on mercurial-induced reduction of 45Ca2+ uptake in isolated nerve terminals of the rat.
The goal of this study was to compare the ability of neurotoxic mercurials which differ in ionic charge and/or lipophilicity to block nerve-terminal calcium channels. To do so, we examined the acute effects of methyl mercury (MeHg+), ethyl mercury (EtHg+), inorganic mercury (Hg2+), dimethyl mercury (Me2Hg), p-chloromercuribenzoate (PCMB), and p-chloromercuriphenyl-sulfonate (PCMBS-) (10-1000 microM) on 45Ca2+ flux into rat forebrain synaptosomes at rest and during depolarization. Basal (depolarization-independent) entry of 45Ca2+ was measured during 10-sec exposure to mercurials in solutions containing 5 mM KCl. Concentrations of 50, 100, 250, 500, and 1000 microM of Hg2+, MeHg+, and EtHg+ reduced basal influx of 45Ca2+. PCMB reduced basal influx at concentrations of 10, 50, and 100 microM, but increased influx at 1000 microM. PCMBS- and (Me)2Hg had no effect on basal flux at any concentration tested. Uptake of 45Ca2+ was measured after 1 sec of K(+)-induced depolarization (41.25 mM) to determine influx through voltage-dependent Ca2+ channels ("fast" phase) or during the last 10 sec of a 20-sec period of depolarization for uptake associated with a reversed Na+/Ca2+ exchanger and a residual noninactivating Ca2+ channel component ("slow" phase). Fast and slow components of 45Ca2+ uptake into synaptosomes were blocked in a concentration-dependent manner by MeHg+, EtHg+, and Hg2+. For block of the fast component, the calculated IC50's and confidence intervals were (microM) EtHg+, 92 (82, 102); Hg2+, 155 (149, 161); and MeHg+, 196 (120, 272). IC50's and the confidence intervals for the slow component of uptake were (microM) Hg2+, 49 (43, 55); MeHg+, 72 (67, 77); and EtHg+, 147 (142, 152). In contrast, Me2Hg, PCMB, and PCMBS- (10-1000 microM) caused no appreciable reduction in either phase of 45Ca2+ uptake. Increasing [Ca2+]e was unable to overcome the block induced by MeHg+ and EtHg+ (100 microM) on either phase of 45Ca2+ uptake into synaptosomes. Likewise, increasing [Ca2+]e failed to overcome block of the slow component by Hg2+ (100 microM). Increasing [Ca2+]e was able to overcome, in part, block of the fast phase induced by Hg2+ (100 microM) although the percentage of reversal was not statistically significant. The magnitude of block of 45Ca2+ uptake increased as a function of increasing [K+]e for MeHg+ and EtHg+, suggesting the block to be voltage-dependent. Thus, mercurials of dissimilar charge and lipophilicity affect synaptosomal Ca2+ uptake differentially.(ABSTRACT TRUNCATED AT 400 WORDS)