Targeting and insertion of C-terminally anchored proteins to the mitochondrial outer membrane is specific and saturable but does not strictly require ATP or molecular chaperones.
A distinct class of proteins contain a C-terminal membrane anchor and a cytoplasmic functional domain. A subset of these proteins is targeted to the mitochondrial outer membrane. Here, to probe for the involvement of a saturable targeting mechanism for this class of proteins, and to elucidate the roles of chaperone proteins and ATP, we have utilized an in vitro targeting system consisting of in vitro-synthesized proteins and isolated mitochondria. To establish the specificity of targeting we have used a closely related protein pair. VAMP-1A and VAMP-1B are splice variants of the vesicle-associated membrane protein/synaptobrevin-1 (VAMP-1) gene. In intact cells VAMP-1B is targeted to mitochondria whereas VAMP-1A is targeted to membranes of the secretory pathway, yet these isoforms differ by only five amino acids at the extreme C-terminus. Here we demonstrate that, in vitro, VAMP-1B is imported into both intact mitochondria and mitochondrial outer-membrane vesicles with a 15-fold greater efficiency than VAMP-1A. We generated and purified bacterially expressed fusion proteins consisting of the C-terminal two-thirds of VAMP-1A or -1B proteins fused to glutathione S-transferase (GST). Using these fusion proteins we demonstrate that protein targeting and insertion is saturable and specific for the VAMP-1B membrane anchor. To elucidate the role of cytosolic chaperones on VAMP-1B targeting, we also used the purified, Escherichia coli-derived fusion proteins. (33)P-Labelled GST-VAMP-1B(61-116), but not GST-VAMP-1A(61-118), was efficiently targeted to mitochondria in a chaperone-free system. Thus the information required for targeting is contained within the targeted protein itself and not the chaperone or a chaperone-protein complex, although chaperones may be required to maintain a transport-competent conformation. Moreover, ATP was required for transport only in the presence of cytosolic chaperone proteins. Therefore the ATP requirement of transport appears to reflect the participation of chaperones and not any other ATP-dependent step. These data demonstrate that targeting of C-terminally anchored proteins to mitochondria is sequence specific and mediated by a saturable mechanism. Neither ATP nor chaperone proteins are strictly required for either specific targeting or membrane insertion.


TFPP: an SVM-based tool for recognizing flagellar proteins in Trypanosoma brucei.
Trypanosoma brucei is a unicellular flagellated eukaryotic parasite that causes African trypanosomiasis in human and domestic animals with devastating health and economic consequences. Recent studies have revealed the important roles of the single flagellum of T. brucei in many aspects, especially that the flagellar motility is required for the viability of the bloodstream form T. brucei, suggesting that impairment of the flagellar function may provide a promising cure for African sleeping sickness. Knowing the flagellum proteome is crucial to study the molecular mechanism of the flagellar functions. Here we present a novel computational method for identifying flagellar proteins in T. brucei, called trypanosome flagellar protein predictor (TFPP). TFPP was developed based on a list of selected discriminating features derived from protein sequences, and could predict flagellar proteins with ∼92% specificity at a ∼84% sensitivity rate. Applied to the whole T. brucei proteome, TFPP reveals 811 more flagellar proteins with high confidence, suggesting that the flagellar proteome covers ∼10% of the whole proteome. Comparison of the expression profiles of the whole T. brucei proteome at three typical life cycle stages found that ∼45% of the flagellar proteins were significantly changed in expression levels between the three life cycle stages, indicating life cycle stage-specific regulation of flagellar functions in T. brucei. Overall, our study demonstrated that TFPP is highly effective in identifying flagellar proteins and could provide opportunities to study the trypanosome flagellar proteome systematically. Furthermore, the web server for TFPP can be freely accessed at http:/wukong.tongji.edu.cn/tfpp.


Retinal Vascular Morphological Changes in Patients with Extremely Severe Obstructive Sleep Apnea Syndrome.
Obstructive sleep apnea syndrome (OSAS) has been shown to generate hypertension and endothelial dysfunction. Retinal vessel is the only vessel that can be observed directly and noninvasively; retinal vascular abnormalities can serve as a predictive marker for the occurrence, clinical course, and prognosis of cardiovascular and cerebrovascular diseases. The objective of this study was to identify the effect of OSAS severity on the morphological changes of retinal vessels. Adult patients complained of snoring were included in this study. The patients' general information, polysomnography, and fundus photography parameters including central retinal artery equivalent (CRAE), central retinal vein equivalent (CRVE), and arteriole-to-venule ratio (AVR) were collected. Patients were divided into four groups according to their apnea-hypopnea index (AHI) results: Group I, AHI ≤5/h; Group II, 5/h < AHI ≤30/h; Group III, 30/h < AHI ≤60/h; and Group IV, AHI> 60/h. A total of 133 patients were included in this study with 111 males (83.5%) and 22 females (16.5%). Mean age was 41.6 ± 9.9 years, and the mean body mass index was 28.1 ± 4.0 kg/m2. AHI ranged between 0 and 130.8/h with a mean of 39.1 ± 30.7/h. There were 24, 34, 35, and 40 patients in Group I, Group II, Group III, and Group IV, respectively. Significant differences were found for AHI (F = 388.368, P< 0.001), minimal pulse oxygen saturation (F = 91.902, P< 0.001), and arousal index (F = 31.014, P< 0.001) among four groups; no significant differences were found for CRAE (F = 0.460, P = 0.599) and CRVE (F = 0.404, P = 0.586) among groups; there were significant differences for AVR between Group I and Group IV (63.6 ± 5.1% vs. 67.2 ± 5.5%, P = 0.010) Group II and Group IV (64.5 ± 6.0% vs. 67.2 ± 5.5%, P = 0.030), and Group III and Group IV (64.7 ± 4.1% vs. 67.2 ± 5.5%, P = 0.043). A main group-by-AHI effect was found on the AVR: patients with higher AHI showed higher AVR results (r = 0.225, P = 0.009). Multivariate logistic regression analysis was used for multi-variable factors. A group-by-age effect was found on the AVR: younger patients showed higher AVR results (β = -0.001, P = 0.020). This study indicated that increased AVR of retinal vessel can be observed in extremely severe OSAS patients. For patients with OSAS, retinal vascular abnormalities may become an early indication for further cardiovascular abnormalities.


PerR vs OhrR: selective peroxide sensing in Bacillus subtilis.
Bacteria adapt to elevated levels of reactive oxygen species (ROS) by increasing the expression of detoxification enzymes and repair proteins. These defences are regulated by transcription factors that activate specific genes in response to ROS. In Bacillus subtilis, the adaptive response to peroxide stress is mainly under control of three proteins: sigma(B), PerR and OhrR. sigma(B) is a general stress response transcription factor. PerR is a dimeric zinc protein with a regulatory site that coordinates either a Fe(2+) or a Mn(2+) metal ion. In the presence of iron, PerR mediates strong induction of the perR regulon in response to hydrogen peroxide (H(2)O(2)). In contrast to PerR, the OhrR protein is weakly activated by H(2)O(2) but it shows a much higher reactivity for organic hydroperoxides. OhrR controls the expression of a thiol-dependent peroxidase that reduces organic hydroperoxides into their corresponding alcohols. In this review we emphasis peroxide sensing mechanisms for both proteins, focusing on recent biochemical and structural data.


Gastric emptying of liquids in the dog: serial test meal and modified emptying-time techniques.
A serial test meal technique was used in the dog to evaluate the gastric emptying of a liquid test meal. On separate days, a phenol red test meal was placed in the stomach and completely aspirated at different times. A composite representation of the gastric emptying pattern was obtained for each dog, and a set of normative data was generated. Three emptying determinants (emptying time, half-life, and starting index) were calculated. The basic pattern of canine gastric emptying was linear and declined exponentially or as a function of the square root of the gastric volume. A modified emptying-time technique (METT) correlated well with the serial test meal and a set of normative values was determined. The METT was performed by aspirating test meal from the stomach 30 minutes after instillation. The METT was a simple and inexpensive technique that could be performed in 1 short testing period. With the METT, the half-life of gastric emptying was calculated, using the log of the percentage of retention of test meal; gastric emptying time was calculated, using the square root of the percentage of retention of test meal. Acetylpromazine was shown not to alter gastric emptying of a liquid test meal.


Enhanced binding of peptide antigen to purified class II major histocompatibility glycoproteins at acidic pH.
Helper T lymphocytes recognize peptide antigens stably associated with class II major histocompatibility complex (MHC) glycoproteins on the surface of antigen-presenting cells and serve to regulate a wide variety of immune responses. A previous study from our laboratory had demonstrated that the functional association of various peptide antigens with the antigen-presenting cell membrane was increased at pH 5 as compared to pH 7, consistent with the potential role of acidic endosomal compartments in antigen processing. The mechanism for this effect was not determined. In the present study, assays using purified class II glycoprotein were used to further define this mechanism. The potential requirement for pH-dependent interactions involving non-MHC membrane components was excluded in functional assays with purified class II reconstituted in artificial membranes containing only neutral phospholipids and cholesterol. The association of HEL(104-120) with I-Ed, and OVA(323-339) with I-Ad, was increased at pH 5, as measured by activation of specific T cell hybridomas. An enzyme immunoassay was developed to measure the binding of biotin-labeled peptides to purified class II in detergent micelles. The pH dependence of binding paralleled our previous functional results. Optimum binding of biotin-HEL(104-120) to I-Ed was observed at pH approximately 4.5, whereas maximum binding of biotin-Myo(106-118) to I-Ad occurred at pH approximately 5.5. The latter peptide also bound weakly to I-Ed, but with a pH dependence similar to that observed using HEL(104-120). Further experiments with biotin-HEL(104-120)/I-Ed indicated that both the apparent affinity and the apparent concentration of peptide-binding sites are increased as hydrogen ion concentration is increased from pH 7 to pH 5. The effect of pH in this range was largely reversible and was not associated with a change in peptide dissociation that could be measured with our assay system. Binding was not inhibited in the presence of 1.5 M NaCl, suggesting that electrostatic interactions between HEL(104-120) and I-Ed are not essential for binding. It is proposed that protonation of a critical group(s) in the class II molecule regulates its capacity to form stable complexes with peptide. However, this effect alone does not fully account for the rapid kinetics of peptide binding observed in experiments with intact antigen-presenting cells.


Paying for quality: providers' incentives for quality improvement.
Paying health care providers to meet quality goals is an idea with widespread appeal, given the common perception that quality of care in the United States remains unacceptably low despite a decade of benchmarking and public reporting. There has been little critical analysis of the design of the current generation of quality incentive programs. In this paper we examine public reports of paying for quality over the past five years and assess each of the identified programs in terms of key design features, including the market share of payers, the structure of the reward system, the amount of revenue at stake, and the targeted domains of health care quality.


Rejuvenating the Brain With Chronic Exercise Through Adult Neurogenesis.
The aging brain presents a general decline in plasticity that also affects hippocampal neurogenesis. Besides the well-known reduction in the rate of neuronal generation, development of new neurons is largely delayed in the aging brain. We have recently shown that this slow development is accelerated when middle-aged mice perform voluntary exercise in a running wheel. It is unclear whether the effects of exercise on neurogenic plasticity are persistent in time in a manner that might influence neuronal cohorts generated over an extended time span. To clarify these issues, we examined the effects of exercise length in 3-week-old neurons and found that their development is accelerated only when running occurs for long (3-4 weeks) but not short periods (1 week). Furthermore, chronic running acted with similar efficiency on neurons that were born at the onset, within, or at the end of the exercise period, lasting until 3 months. Interestingly, no effects were observed on neurons born 1 month after exercise had ended. Our results indicate that multiple neuronal cohorts born throughout the exercise span integrate very rapidly in the aging brain, such that the effects of running will accumulate and expand network assembly promoted by neurogenesis. These networks are likely to be more complex than those assembled in a sedentary mouse due to the faster and more efficient integration of new neurons.


Maghemite nanocrystal impregnation by hydrophobic surface modification of mesoporous silica.
Here, we report the design of a hybrid inorganic/organic mesoporous material through simultaneous pore engineering and hydrophobic surface modification of the intramesochannels to improve the uptake of superparamagnetic maghemite nanocrystals via impregnation techniques. The mesoporous material of the SBA-15 type was functionalized in situ with thiol organo-siloxane groups. Restricting the addition of the thiol organo-siloxane to 2 mol % yielded an inorganic/organic hybrid material characterized by large pores and a well-ordered hexagonal p6mm mesophase. The hydrophobic surface modification promoted the incorporation of 7.5 nm maghemite (gamma-Fe2O3) nanocrystals, prepared through temperature-controlled decomposition of iron pentacarbonyl in organic solvents. The hydrophobic, oleic acid capped superparamagnetic maghemite nanocrystals were incorporated into the porous network via wet impregnation from organic suspensions. Combining diffraction, microscopy, and adsorption data confirmed the uptake of the nanocrystals within the intramesochannels of the silica host. Magnetization dependencies on magnetic field at different temperatures show a constriction in the loop around the origin, which indicates immobilization of maghemite nanocrystals inside the thiol-functionalized silica host.


Association between remnant lipoprotein cholesterol levels and non-alcoholic fatty liver disease in adolescents.
Remnant lipoprotein cholesterol (RLP-C) is an atherogenic lipid profile associated with non-alcoholic fatty liver disease (NAFLD) and cardiovascular disease (CVD). With increased rates of CVD seen in adults with NAFLD, RLP-C has the potential to identify individuals with NAFLD who are at increased risk of CVD. This study examined in adolescents sex-different associations among RLP-C, NAFLD, and cardiometabolic risk factors, and whether RLP-C is associated with NAFLD beyond traditional cardiometabolic risk factors. Adolescents in the Raine Study had anthropometry, clinical, biochemistry and arterial stiffness measurements recorded at 17 years of age. Fatty liver, subcutaneous and visceral adipose thickness were assessed using abdominal ultrasound. Relationships among RLP-C, NAFLD, liver biochemistry, insulin resistance, adipokines, adiposity and arterial stiffness were assessed. NAFLD was diagnosed in 15.1% (19.6% females and 10.7% males) of adolescents. Increasing RLP-C levels were associated with increasing severity of hepatic steatosis and metabolic syndrome. Adolescents with NAFLD and serum RLP-C levels in the highest quartile compared with the lowest quartile, had higher serum leptin, homeostatic model assessment of insulin resistance (HOMA-IR), high-sensitivity C-reactive protein, low-density-lipoprotein cholesterol, triglycerides, BMI, subcutaneous and visceral adipose thickness, systolic blood pressure and arterial stiffness, but lower adiponectin and high-density-lipoprotein cholesterol. Using multivariable logistic regression, RLP-C in the lowest quartile compared with the highest quartile was associated with 85% lower odds of NAFLD in males and 55% in females, after adjusting for waist circumference, leptin, ALT, adiponectin and HOMA-IR. There is an association between RLP-C and NAFLD beyond traditional risk factors of adiposity and insulin resistance in adolescents. Although raised serum RLP-C levels were associated with the severity of hepatic steatosis and markers of cardiometabolic risk, lower serum RLP-C might reflect reduced cardiovascular risk. Remnant lipoprotein cholesterol (RLP-C) is a part of the blood cholesterol that is linked with heart disease and non-alcoholic fatty liver disease (NAFLD) in adults. In the Raine Study, teenagers with high RLP-C levels had more severe fat accumulation in their liver. Thus, RLP-C might be the hidden link between NAFLD and future risk of heart disease.


Magnetite nanoparticles-based hydroxyl radical scavenging activity assay of antioxidants using N, N-dimethyl-p-phenylenediamine probe.
Excessive amounts of reactive oxygen species (ROS), unless counterbalanced by antioxidants, can cause cellular damage under oxidative stress conditions; therefore, antioxidative defenses against ROS must be measured. With the development of nanotechnology, nanoparticles have found numerous applications in science, health, and industries. Magnetite nanoparticles (Fe 3 O 4 :MNPs) have attracted attention because of their peroxidase-like activity. In this study, hydroxyl radicals (•OH) generated by MNPs-catalyzed degradation of H 2 O 2 converted the N,N-dimethyl-p-phenylenediamine (DMPD) probe into its colored DMPD•+ radical cation, which gave an absorbance maximum at λ = 553 nm. In the presence of antioxidants, •OH was partly scavenged by antioxidants and produced less DMPD• + , causing a decrease in the 553 nm-absorbance. Antioxidant concentrations were calculated with the aid of absorbance differences between the reference and sample solutions. The linear working ranges and trolox equivalent antioxidant capacity coefficients of different classes of antioxidants were determined by applying the developed method. In addition, binary and ternary mixtures of antioxidants were tested to observe the additivity of absorbances of mixture constituents. The method was applied to real samples such as orange juice and green tea. Student t-test, F tests, and the Spearman's rank correlation coefficient were used for statistical comparisons.


Increased aortic PGI2 and plasma lyso-PAF in the unclipped one-kidney hypertensive rat.
Previous studies have implicated vasodilatory prostaglandins (PGs) in the reversal of hypertension following unclipping in the one-kidney, one-clip (1K,1C) hypertensive rat. The capacity of the aorta to synthesize prostacyclin (PGI2) was compared in clipped (group A, n = 9), unclipped (group B, n = 8 and group D, n = 9), and sham-unclipped (group C, n = 9) 1K,1C hypertensive rats. The involvement of platelet-activating factor (PAF), a potent renal antihypertensive phospholipid, in the reversal of renal clip hypertension was also examined. Hypertensive rats [systolic blood pressure (BP) greater than 180 mmHg] were fed a synthetic diet for 4 wk, after which group A was killed immediately, group C was sham-unclipped, and groups B and D unclipped and killed 24 h later. Blood was drawn for the measurement of plasma lyso-PAF (the precursor of PAF) and the aorta removed for determination of 6-ketoprostaglandin F1 alpha (6-keto-PGF1 alpha, the stable hydrolysis product of PGI2). BP fell substantially in the unclipped rats (groups B and D) but did not change in the sham-unclipped rats (group C). Mean aortic 6-keto-PGF1 alpha was increased in the unclipped groups [group B, 15.4 +/- 2.4 (SE) ng/mg; group D, 10.8 +/- 2 ng/mg] compared with group A (7.7 +/- 1 ng/mg) and group C (7.1 +/- 1 ng/mg) (H = 13.74, P less than 0.01). Plasma lyso-PAF was also significantly increased in the unclipped (group D, 261 +/- 26 ng/ml) vs. the sham-unclipped group (group C, 211 +/- 23 ng/ml, P less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)


Sequential hypothesis testing with spatially correlated presence-absence data.
A pest management decision to initiate a control treatment depends upon an accurate estimate of mean pest density. Presence-absence sampling plans significantly reduce sampling efforts to make treatment decisions by using the proportion of infested leaves to estimate mean pest density in lieu of counting individual pests. The use of sequential hypothesis testing procedures can significantly reduce the number of samples required to make a treatment decision. Here we construct a mean-proportion relationship for Oligonychus perseae Tuttle, Baker, and Abatiello, a mite pest of avocados, from empirical data, and develop a sequential presence-absence sampling plan using Bartlett's sequential test procedure. Bartlett's test can accommodate pest population models that contain nuisance parameters that are not of primary interest. However, it requires that population measurements be independent, which may not be realistic because of spatial correlation of pest densities across trees within an orchard. We propose to mitigate the effect of spatial correlation in a sequential sampling procedure by using a tree-selection rule (i.e., maximin) that sequentially selects each newly sampled tree to be maximally spaced from all other previously sampled trees. Our proposed presence-absence sampling methodology applies Bartlett's test to a hypothesis test developed using an empirical mean-proportion relationship coupled with a spatial, statistical model of pest populations, with spatial correlation mitigated via the aforementioned tree-selection rule. We demonstrate the effectiveness of our proposed methodology over a range of parameter estimates appropriate for densities of O. perseae that would be observed in avocado orchards in California.


Brain capillary telangiectasias: from normal variants to disease.
Brain capillary telangiectasias (BCTs) are small, dilated capillary networks in the brain that are most commonly asymptomatic. Though rare, symptomatic cases of BCTs have been reported, and it is therefore important to understand the nature of these lesions in order to facilitate proper recognition. Relative to other intracranial vascular malformations, updated information on the various epidemiologic, radiographic, and pathologic features of BCTs within the published literature may be inadequate. We searched the PubMed database for prior reports of symptomatically-manifested BCTs. Moreover, Google Scholar and PubMed were searched in order to review current epidemiologic, radiographic, pathologic, and pathogenetic features of BCTs. Forty-eight published studies were included for a total of 99 individual cases of BCTs with symptomatic manifestations. Thirty-three symptomatic BCTs were hemorrhagic in nature, while 66 were non-hemorrhagic. The mean age at presentation of hemorrhagic lesions was 25.5 years, and the most common location was the supratentorial CNS (54.5%) with motor disturbance representing the most commonly encountered presenting symptom (26.1%). 15.2% of hemorrhagic lesions were treated with surgical removal. In non-hemorrhagic lesions, the mean age at presentation was 39.8 years with the pons being the most common lesion location (78.5%) and headache being the most common presenting symptom (22.2%). 12.1% of patients with non-hemorrhagic lesions were treated with surgical removal. Despite their rarity, symptomatic BCTs, both hemorrhagic and non-hemorrhagic, can cause devastating neurological sequelae, potentially through multiple mechanisms. The large majority of these lesions do not require intervention, though surgical removal has been achieved with good outcome in select cases. Further documentation of symptomatic manifestations with or without surgical intervention is vital in order to further understand the clinical, surgical, and pathogenic implications of these less-appreciated vascular malformations.


A syngeneic monoclonal antibody to murine Meth-A sarcoma (HepSS-1) recognizes heparan sulfate glycosaminoglycan (HS-GAG): cell density and transformation dependent alteration in cell surface HS-GAG defined by HepSS-1.
We have isolated a syngeneic monoclonal antibody (HepSS-1) reactive to a murine methylcholanthrene-induced fibrosarcoma, Meth-A. HepSS-1 also bound to a wide variety of established and fresh normal cells derived from not only mice but also other species such as human, monkey, rat, hamster, and chicken. Immunoprecipitation of surface iodinated Meth-A cell extract with HepSS-1, as well as Sepharose 4B gel chromatography of Meth-A cell extract and detection of antigens recognized by HepSS-1 by a sandwich-type radioimmunoassay revealed that the HepSS-1 antigens were composed of several molecular species, with one as large as approximately 10(6) daltons. The following evidence indicates that HepSS-1 specifically recognizes an epitope present in heparan sulfate glycosaminoglycan (HS-GAG). First, treatment of Meth-A cells with heparitinase or heparinase, but not with chondroitinase ABC or hyaluronidase, resulted in the loss of HepSS-1 binding. Second, HS-GAG but not seven other types of GAG (hyaluronic acid, heparin, chondroitin, chondroitin 4-sulfate, chondroitin 6-sulfate, dermatan sulfate, and keratan sulfate) inhibited HepSS-1 binding to Meth-A cells. Third, HepSS-1 bound with HS-GAG but not with the seven other types of GAG. From the binding analysis of HepSS-1 to various modified HS-GAG and whale omega-heparin, it is additionally suggested that HepSS-1 recognizes an epitope closely related to O-sulfated and N-acetylated glucosamine. We found that NIH 3T3 cells expressed more HepSS-1 epitopes at a low cell density than at confluency and in G2 + M than in G1, whereas NIH 3T3 cells transformed with Kirsten-ras oncogene or SV-40 expressed high levels of HepSS-1 epitopes and ceased to show the density-dependent change in the amount of HepSS-1 epitopes. These observations were also reproduced by using NIH 3T3 cells transformed with a temperature sensitive Kirsten murine sarcoma virus maintained at permissive and non-permissive temperatures. Thus HepSS-1 is a first monoclonal antibody to HS-GAG and seems to be useful to elucidate changes in cell surface HS-GAG in normal cell growth and cell transformation.


Ultrafast excitation energy transfer in a benzimidazole-naphthopyran donor-acceptor dyad.
The excited-state dynamics of a donor-acceptor dyad composed of 1-propyl-2-pyridinyl-benzimidazole (PPBI) as donor and the photochromic molecular switch diphenylnaphthopyran (DPNP) as acceptor linked via an ester bridge has been investigated by a combination of static and time-resolved spectroscopies and quantum chemical calculations. The UV absorption spectrum of the dyad is virtually identical to the sum of the spectra of its individual constituents, indicating only weak electronic coupling between the donor and acceptor in the electronic ground state. After selective photoexcitation of the PPBI chromophore in the dyad at λpump = 310 nm, however, a fast electronic energy transfer (EET) from the donor to the acceptor is observed, by which the lifetime of the normally long-lived excited state of PPBI is reduced to a few ps. Enabled by the EET, the acceptor switches from its ring-closed naphtopyran form to its ring-opened merocyanine form. The singular value decomposition-based global analyses of the measured femtosecond time-resolved transient absorption spectra of the dyad and its two building blocks as reference compounds allowed us to determine a value for the EET time constant in the dyad of τ = 2.90 ± 0.60 ps. For comparison, Förster theory predicts characteristic FRET times between 1.2 ps ≤ τ ≤ 4.2 ps, in good agreement with the experimental result.


Minimum number of bacteria needed for antigen detection by counterimmunoelectrophoresis: in vivo and in vitro studies.
Threshold concentrations of Streptococcus pneumoniae type 3, Haemophilus influenzae type b, and Streptococcus sp. group B type Ib required for positive counterimmunoelectrophoresis reactions were determined in vivo and in vitro. Animals were infected intraperitoneally with various concentrations of microorganisms: adult mice with S. pneumoniae, suckling rats with H. influenzae, and 3-week-old mice with Streptococcus sp. group B. At 24 h after infection a minimum blood concentration of 10(3) colony-forming units (CFU)/ml was needed for S. pneumoniae or H. influenzae before antigen was detected in the serum. A minimum concentration of 10(6) CFU/ml was needed for Streptococcus sp. group B at 10 h after infection. Larger threshold concentrations (10(4) CFU/ml for S. pneumoniae, 10(5) CFU/ml for H. influenzae, and 10(7) CFU/ml for Streptococcus) were required in broth-grown cultures before cell-free antigens could be demonstrated by counterimmunoelectrophoresis in the medium. Marked levels of antigen release by group B streptococci were observed as the cultures entered early stationary phase. This study provides evidence of a long-accepted, though poorly substantiated, hypothesis that a threshold concentration of microorganism is necessary before counterimmunoelectrophoresis reactions become positive. Counterimmunoelecrophoresis results for clinical specimens should be interpreted cautiously in light of this evidence.


Systematic review and meta-analysis: islet autotransplantation after pancreatectomy for minimizing diabetes.
Islet autotransplantation (IAT) may decrease the morbidity and mortality of postpancreatectomy diabetes mellitus. The current systematic review and meta-analysis examined the rate of insulin independence (II) and mortality after IAT post-total (TP) or partial pancreatectomy (PP). Ovid MEDLINE, EMBASE, Web of Science, SCOPUS and reference lists were searched until 31 January 2011. Eligible studies enrolled adult patients with IAT post-TP or PP, regardless of study design, sample size and language. Two investigators identified eligible studies and extracted data independently. From each study, 95% confidence intervals (CIs) were estimated and pooled using random effects meta-analysis. Fifteen observational studies were eligible (11 IAT post-TP, two post-PP and two including both). The II rates for IAT post-TP at last follow-up and transiently during the study were 4·62 per 100 person-years (95% CI: 1·53-7·72) and 8·34 per 100 person-years (95% CI: 3·32-13·37), respectively. In the later group, patients achieved transient II lasting 15·57 months (95% CI: 10·35-20·79). The II rate at last follow-up for IAT post-PP was 24·28 per 100 person-years (95% CI: 0·00-48·96). Whereas the 30-day mortality for IAT post-TP and post-PP was 5% (95% CI: 2-10%) and 0, respectively, the long-term mortality was 1·38 per 100 person-years (95% CI: 0·66-2·11) and 0·70 per 100 person-years (95% CI: 0·00-1·80) respectively. IAT postpancreatectomy offers some patients a chance for insulin independence. Better data reporting are essential to establish the risks and benefits of IAT after pancreatic surgery.


Comprehensive Characterization of COVID-19 Patients with Repeatedly Positive SARS-CoV-2 Tests Using a Large U.S. Electronic Health Record Database.
In the absence of genome sequencing, two positive molecular tests for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) separated by negative tests, prolonged time, and symptom resolution remain the best surrogate measure of possible reinfection. Using a large electronic health record database, we characterized clinical and testing data for 23 patients with repeatedly positive SARS-CoV-2 PCR test results ≥60 days apart, separated by ≥2 consecutive negative test results. The prevalence of chronic medical conditions, symptoms, and severe outcomes related to coronavirus disease 19 (COVID-19) illness were ascertained. The median age of patients was 64.5 years, 40% were Black, and 39% were female. A total of 83% smoked within the prior year, 61% were overweight/obese, 83% had immunocompromising conditions, and 96% had ≥2 comorbidities. The median interval between the two positive tests was 77 days. Among the 19 patients with 60 to 89 days between positive tests, 17 (89%) exhibited symptoms or clinical manifestations consistent with COVID-19 at the time of the second positive test and 14 (74%) were hospitalized at the second positive test. Of the four patients with ≥90 days between two positive tests (patient 2 [PT2], PT8, PT14, and PT19), two had mild or no symptoms at the second positive test and one, an immunocompromised patient, had a brief hospitalization at the first diagnosis, followed by intensive care unit (ICU) admission at the second diagnosis 3 months later. Our study demonstrated a high prevalence of compromised immune systems, comorbidities, obesity, and smoking among patients with repeatedly positive SARS-CoV-2 tests. Despite limitations, including a lack of semiquantitative estimates of viral load, these data may help prioritize suspected cases of reinfection for investigation and continued surveillance. IMPORTANCE The comprehensive characterization of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) testing and clinical data for patients with repeatedly positive SARS-CoV-2 tests can help prioritize suspected cases of reinfection for investigation in the absence of genome sequencing data and for continued surveillance of the potential long-term health consequences of SARS-CoV-2 infection.


Impact of trauma system structure on injury outcomes: a systematic review protocol.
Injury represents one of the greatest public health challenges of our time with over 5 million deaths and 100 million people temporarily or permanently disabled every year worldwide. The effectiveness of trauma systems in decreasing injury mortality and morbidity has been well demonstrated. However, the organisation of trauma care varies significantly across trauma systems and we know little about which components of trauma systems contribute to their effectiveness. The objective of the study described in this protocol is to systematically review evidence of the impact of trauma system components on clinically significant outcomes including mortality, function and disability, quality of life, and resource utilization. We will perform a systematic review of studies evaluating the association between at least one trauma system component (e.g. accreditation by a central agency, interfacility transfer agreements) and at least one injury outcome (e.g. mortality, disability, resource use). We will search MEDLINE, EMBASE, COCHRANE central, and BIOSIS/Web of Knowledge databases, thesis holdings, key injury organisation websites and conference proceedings for eligible studies. Pairs of independent reviewers will evaluate studies for eligibility and extract data from included articles. Methodological quality will be evaluated using elements of the ROBINS-I tool and the Cochrane risk of bias tool for non-randomized and randomized studies, respectively. Strength of evidence will be evaluated using the GRADE tool. We expect to advance knowledge on the components of trauma systems that contribute to their effectiveness. This may lead to recommendations on trauma system structure that will help policy-makers make informed decisions as to where resources should be focused. The review may also lead to specific recommendations for future research efforts. This protocol was registered with the International Prospective Register of Systematic Reviews (PROSPERO) on 28-06-2016. PROSPERO 2016:CRD42016041336 Available from http://www.crd.york.ac.uk/PROSPERO/display_record.asp?ID=CRD42016041336 .


The role of chloride in deoxycorticosterone hypertension: selective sodium loading by diet or drinking fluid.
To evaluate the role of chloride in the pathogenesis of salt-dependent deoxycorticosterone (DOC) hypertension, we studied young Wistar rats chronically loaded with sodium bicarbonate (NaHCO(3)) or sodium chloride (NaCl) which were administered either in the diet or in the drinking fluid. Selective sodium loading (without chloride) increased blood pressure (BP) in DOC-treated animals only if NaHCO(3) was provided in the diet. In contrast, no significant blood pressure changes were induced by DOC treatment in rats drinking NaHCO(3) solution. Hypernatremia and high plasma osmolality occurred only in rats drinking NaCl or NaHCO(3) solutions. Compared to great volume expansion in NaCl-loaded DOC-treated rats, the degree of extracellular fluid volume expansion (namely of its interstitial fraction) was substantially lower in both NaHCO(3)-loaded groups in which significant hypokalemia was observed. NaHCO(3)-drinking rats without significant blood pressure response to DOC treatment represented the only experimental group in which blood volume was not expanded. In conclusion, our data confirm previous observations that NaHCO(3) loading is less potent in eliciting DOC hypertension than NaCl loading, but blood pressure rise in rats fed NaHCO(3) diet clearly demonstrated that selective sodium loading could potentiate the development of DOC hypertension if NaHCO(3) is offered within the appropriate dietary regimen. The reasons for the failure of NaHCO(3)-drinking rats to elevate blood pressure in response to chronic mineralocorticoid treatment are not obvious. However, the absence of a significant plasma volume expansion together with hypernatremia and increased plasma osmolality suggest a considerable degree of dehydration in these animals which fail to increase their fluid consumption compared to water drinking rats.


Hyphal tip growth in Phytophthora. Gradient distribution and ultrahistochemistry of enzymes.
Germinating cysts and isolated walls from germinating cysts incorporated 14C-UDPG into wall material of which 22.5 and 15% respectively were insoluble in boiling 1 N HCl, indicating that part of the synthetase activity is located in the wall itself. A combination of Urografin and Ficoll density gradients was used to separate various intracellular fractions. A consistent separation of beta-glucanase and UDPG-transferase enriched fractions was achieved. The beta-glucanase fraction contained dictyosome vesicles and fragments along with some plasma membranes. The UDPG-transferase fraction was relatively rich in membranes resembling rough and smooth ER. The results suggest the two enzymes are transported to the wall by different intracellular routes, and two types of vesicle may be involved. Alkaline phosphatase, beta-glucosidase and acid phosphatase were found extracellularly and their distribution in density gradients determined. The results of histochemical staining for acid phosphatase, alkaline phosphatase and polysaccharide are described and compared with the biochemical data. beta-1,3-glucanase, found intra- and extracellularly, induced distorted growth of germ tubes and also removed most of the apical wall when added to the incubation medium. None of these responses were observed with cellulase. Determinations of the osmotic pressure of germinating cysts and incubation medium revealed that the turgor of germinating cysts amounts to about 1.8 at under the conditions used.


Broad-range survey of tick-borne pathogens in Southern Germany reveals a high prevalence of Babesia microti and a diversity of other tick-borne pathogens.
Abstract Ticks harbor numerous pathogens of significance to human and animal health. A better understanding of the pathogens carried by ticks in a given geographic area can alert health care providers of specific health risks leading to better diagnosis and treatments. In this study, we tested 226 Ixodes ricinis ticks from Southern Germany using a broad-range PCR and electrospray ionization mass spectrometry assay (PCR/ESI-MS) designed to identify tick-borne bacterial and protozoan pathogens in a single test. We found 21.2% of the ticks tested carried Borrelia burgdorferi sensu lato consisting of diverse genospecies; a surprisingly high percentage of ticks were infected with Babesia microti (3.5%). Other organisms found included Borrelia miyamotoi, Rickettsia helvetica, Rickettsia monacensis, and Anaplasma phagocytophilum. Of further significance was our finding that more than 7% of ticks were infected with more than one pathogen or putative pathogen.


Epidemiology & attributing factors for chronic kidney disease: Finding from a case-control study in Odisha, India.
Chronic kidney disease (CKD) is one of the leading causes of mortality in developing countries, however, evidence from some geographical areas of India is scantly available on its risk factors. Other than diabetes and hypertension, several personal and environmental factors are also associated with CKD. A population-based case-control study was conducted over a period of 12 months in two high CKD reporting districts of Odisha, India. A total of 236 participants, 1:2 age- and sex-matched cases (83):controls (153), were included. Various factors were modelled with univariate and multivariable conditional logistic regression and analyzed using the Bayesian method in STATA SE v.12. Among the study cases, about 81 per cent were male and about 25 per cent were aged <40 yr. CKD-associated risk factors were hypertension for more than five years [adjusted odds ratio (aOR)=4.24; 95% credible interval: 1.23-10.05], scheduled tribe/caste (aOR=2.81; 1.09-5.95), use of tube well/bore well water for drinking (aOR=1.21; 1.02-1.43), consumption of locally made alcohol (aOR=1.09; 1.02-1.17) and eating red meat and vegetarian diet with (aOR=1.24; 1.12-1.39) and (aOR=1.09; 1.04-1.14), respectively. The presence of CKD among younger age groups and association of multiple factors including personal and environmental suggest for more research to establish the cause and effect relation of these factors.


Correlation of a polymorphism in the interleukin-1 receptor antagonist gene with hepatic fibrosis in Japanese alcoholics.
The aim of this study was to determine whether there is any association between interleukin-1 receptor antagonist (IL1-Ra) genotype and alcoholic liver disease. The IL1-Ra genotype was assessed in 102 Japanese male alcoholic liver disease patients and 46 healthy subjects by polymerase chain reaction with leukocyte DNA. The distribution of IL1-Ra genotype and the allelic frequencies in Japanese healthy subjects are both significantly different from that previously reported in Caucasians (A1/A1 genotype: 95.7% in Japanese vs. 54.0% in Caucasians, p < 0.001; A1 allele: 97.8% vs. 73.4%, p < 0.001). The frequency of A1 heterozygotes tended to be higher in Japanese alcoholics with fibrosis, compared with those without fibrosis (14.9% vs. 2.9%). Furthermore, within the fibrotic groups, cumulative alcohol intake was significantly lower in A1 heterozygotes than in the A1 homozygotes (877 +/- 118 kg vs. 1369 +/- 90 kg,p < 0.05). In conclusion, a genetic polymorphism in the IL1-Ra gene may influence the risk of developing hepatic fibrosis in Japanese alcoholics. The same study should be conducted in Caucasian patients having more frequency of IL1-Ra A1 heterozygotes.


Models for biological ion exchangers. II. Solvent selectivity in DMSO-water-Dowex 50W.
Dimethylsulfoxide-water-Dowex 50W-X8 systems are characterized by measurements of solvent selectively and proton magnetic resonance spectra. The H+-, Li+-, NH4+-, NHe4+-, NBu4+-, Mg2+-, Zn2+-, and La3+- forms are studied over a wide range of binary-solvent mole fractions. The relative selectivities for water by the ion exchanger, based on an integrated Kipling parameter, are Zn2+-form (reference) --1.00, Mg2+ --0.43, La3+ --0.38, Li+ +0.17, NMe4+ 0.20, NH4+ 0.31, NBU4+ 0.33, and H+ 0.68, the polyvalent counterions preferring DMSO. All of the ionic forms except the NH4+-form exhibit over much of the mole fraction range separations between the external water and the internal water peaks exceeding 50 Hz, the magnitude of the separation varying with the counter-ion. Comparison of results is facilitated by maintaining a constant ratio between the total number of moles of solvent and the number of equivalents of ions exchanger.


The Antimicrobial Activities of Extract and Compounds Isolated from Brillantaisia lamium.
Brillantaisia lamium is an erect branched herb, which grows to a height of 1.50 m in moist tropical areas, both in full sun and partial shade. In , the aerial part of this plant is used in the treatment of various microbial infections such as skin diseases and infections of urinary tract. The aim of this study was to evaluate the antimicrobial activities of CH(2)Cl(2): MeOH (1:1) extract, fractions and compounds from the aerial part of B. lamium. The plant was dried and extracted by maceration in CH(2)Cl(2): MeOH (1:1 v/v). Structures of the compounds from the CH(2)Cl(2): MeOH (1:1) soluble fraction were determined by spectroscopic methods and compared with published data. The broth micro dilution method was used to evaluate the antimicrobial activities against bacteria and fungal species. Four known compounds: aurantiamide acetate (1), lupeol (2), lespedin (3), sitosterol 3-O-β-D-glucopyranoside (4) and a mixture of sterols: campesterol (5), stigmasterol (6) and β-sitosterol (7) were isolated from CH(2)Cl(2): MeOH (1:1) extract of B. lamium aerial parts. The crude extract, fractions and isolated compounds exhibited both antibacterial and antifungal activities that varied with microorganism (MIC=6.25 - 1000 µg/ml). Compound 3 was the most active (MIC=6.25 - 100 µg/ml) while Staphylococcus aureus, Enterococcus faecalis, Candida tropicalis and Cryptococcus neoformans were the most sensitive to all the tested compounds. The overall results of this study indicate that the CH(2)Cl(2): MeOH (1:1) extract and some of isolated compounds have interesting antimicrobial properties and can be used for the treatment of fungal and bacterial infections.


Nonuniversal impact of cholesterol on membranes mobility, curvature sensing and elasticity.
Biological membranes, composed mainly of phospholipids and cholesterol, play a vital role as cellular barriers. They undergo localized reshaping in response to environmental cues and protein interactions, with the energetics of deformations crucial for exerting biological functions. This study investigates the non-universal role of cholesterol on the structure and elasticity of saturated and unsaturated lipid membranes. Our study uncovers a highly cooperative relationship between thermal membrane bending and local cholesterol redistribution, with cholesterol showing a strong preference for the compressed membrane leaflet. Remarkably, in unsaturated membranes, increased cholesterol mobility enhances cooperativity, resulting in membrane softening despite membrane thickening and lipid compression caused by cholesterol. These findings elucidate the intricate interplay between thermodynamic forces and local molecular interactions that govern collective properties of membranes.


[Traumatic apophyseolysis of the ischiadic tuber].
Traumatic apophyseolysis of the os ischii is a rare injury occurring during sports activities. It mostly affects young men between 14 and 25 years of age. Conservative treatment is often sufficient unless there is a mechanical obstruction or an irritation of the conditioned rupture of the os ischii with subsequent load-dependent complaints.


Falling back on technology mindfully during COVID-19 pandemic: NUMed campus experience.
This article was migrated. The article was marked as recommended. COVID-19 pandemic has forced technology to evolve as the most challenging means of delivering teaching in addressing the cordon sanitaire. Technology has already been transforming most operation in medical education and secured strong footage in medical schools. NUMed, like any other higher educational institution, has started planning and implementing various strategies to ensure teaching is not hampered during this crisis. ReCaps, online tutorials, discussion boards, online seminars, webinars and quizzes were utilized in reflecting on the current situation. The paper assesses and mitigates the immediate change that has breached the boundaries of students and staffs to be engaged with strategic planning and adoption during the COVID-19 situation. This paper illustrates the interrelatedness and navigates across borders to leverage the possible planning and implementing educational technology as the pedagogical means to ensure the quality of medical education is preserved in fact enhanced through these strategies.


Interventions to improve insulin prescribing practice for people with diabetes in hospital: a systematic review.
To conduct a systematic review of literature to identify interventions that are effective in improving insulin prescribing for people with diabetes in the hospital setting. Computerized bibliographic databases were searched for studies published in English that described the effectiveness of interventions to improve insulin prescribing within the hospital setting. Studies were eligible for inclusion if they reported data that compared insulin prescribing practice after an intervention or compared with a control group. Studies were not excluded on the basis of publication date, geographical location or risk of bias assessment. We identified 35 studies for inclusion in the review, including two cluster randomized controlled trials, two cohort studies, and 31 uncontrolled before-after studies. Studies reported a variety of interventions that aimed to increase insulin prescribing accuracy or completeness or decrease the use of discouraged subcutaneous sliding scale insulin regimens. Differences in definition of insulin prescribing error, terminology and common practice based on geographical location was evident, and quality issues with respect to study design and reporting somewhat limited the interpretation of conclusions. Implementing strategies that are sensitive to local context and designed to increase adherence to insulin prescribing guidelines are associated with a reduction in prescribing errors. Future implementation should build on effective approaches including multifaceted interventions involving multiple stakeholders at various institutional levels. Future studies in insulin prescribing errors would benefit from the use of standardized approaches, terminology and outcome measures to enable greater comparison.


Granting immunity to FOP and catching heterotopic ossification in the Act.
The progressive transformation of one organ system into another is a fundamental signature of fibrodysplasia ossificans progressiva (FOP), the most catastrophic form of extraskeletal bone formation in humans. In all affected individuals, FOP is caused by heterozygous missense gain-of-function mutations in Activin receptor A type I (ACVR1), a bone morphogenetic protein (BMP) type I receptor. Loss of autoinhibition of the mutant receptor (mACVR1) results in dysregulated BMP pathway signaling, and is necessary for the myriad developmental features of FOP, but does not appear sufficient to induce the episodic flare-ups that lead to disabling post-natal heterotopic endochondral ossification (HEO) and that are a hallmark of the disease. Post-natal FOP flare-ups strongly implicate an underlying immunological trigger involving inflammation and the innate immune system. Recent studies implicate canonical and non-canonical TGFβ/BMP family ligands in the amplification of mACVR1 signaling leading to the formation of FOP lesions and resultant HEO. BMP and Activin ligands that stimulate mACVR1 signaling also have critical regulatory functions in the immune system. Cross-talk between the morphogenetic and immunological pathways that regulate tissue maintenance and wound healing identifies potential robust therapeutic targets for FOP. Here we review current evidence for an immunological trigger for flare-ups and HEO in FOP, propose a working schema for the pathophysiology of observed phenomena, and highlight outstanding questions under investigation.


Postsynaptic D1 and D2 dopamine receptors are present in rabbit renal and mesenteric arteries.
Measuring adenylate cyclase activity as a biochemical index of dopamine (DA) receptors, it was found that the selective D1 DA receptor agonist, SKF 82526, was able to stimulate the cAMP formation in rabbit renal and mesenteric arteries, an effect blocked by haloperidol and by SCH 23390. The D2 DA receptor agonist, bromocriptine, elicited a concentration-dependent inhibition of adenylate cyclase activity in both arteries of either normal or 6-hydroxydopamine pretreated rabbits, this effect being prevented by (-)-sulpiride but not by (+)-sulpiride. These data indicate that both D1 and D2 postsynaptic DA receptors, associated with stimulation or inhibition of adenylate cyclase activity, are present on the wall of rabbit renal and mesenteric arteries.


Surgical Management of Freiberg Disease by Dorsal Closing Wedge Osteotomy.
The purpose of the study was to assess the efficacy of the dorsal closing wedge osteotomy for the treatment of Freiburg's infraction. Twenty patients with Freiburg's infraction were admitted at our hospital over a period of six years. Patients with a normal plantar contour of the metatarsal head were included. All patients underwent a dorsal closing wedge osteotomy of the metatarsal. The mean Leeds Movement Performance Index (LMPI) score was 84 (range 70-86). The mean metatarsal shortening was 2mm. the passive flexion restriction was 16° and extension restriction was 10°. Also, a strong negative correlation was found between Smillie classification and American Orthopaedic Foot and Ankle Score (AOFAS) final score (r's = -0.85, P < .001). The dorsal closing wedge osteotomy is an efficient and reproducible method for the management of Freiburg's infraction.


Bioluminescence tomography based on the phase approximation model.
A reconstruction method of bioluminescence sources is proposed based on a phase approximation model. Compared with the diffuse approximation, this phase approximation model more correctly predicts bioluminescence photon propagation in biological tissues, so that bioluminescence tomography can accurately locate and quantify the distribution of bioluminescence sources. The compressive sensing (CS) technique is applied to regularize the inverse source reconstruction to enhance numerical stability and efficiency. The numerical simulation and phantom experiments demonstrate the feasibility of the proposed approach.


Bolometer noise: nonequilibrium theory.
New theoretical results for noise in cryogenic bolometers are derived. Johnson noise is reduced by as much as 60% by electrothermal feedback from the bias supply. Phonon noise in the thermal link is reduced by as much as 30% relative to the usual equilibrium formula. Photon noise in the Rayleigh-Jeans limit is computed with attention to the attenuation of the photon correlations in the light beam. Basic results on bolometer responsivity, time constant, and thermal properties are presented in a new and convenient form. Excess 1/f and contact shot noise are also discussed.


Transcriptional control of T4 coliphage-specific genes 30, 42, 43, rIIA, rIIB, and e.
Escherichia coli B/r (suo) was infected, at 30 degrees C, with T4Dam+, T4DamB24-amN82 (I-, 44-, DNA-negative phenotype), and T4DamN134amBL292 (33-, 55-, maturation-defective phenotype). A genetic ('transformation') assay was used to monitor transcription of genes 30 (polynucleotide ligase), 42 (deoxycytidylate hydroxymethylase), 43 (DNA polymerase), rIIA, rIIB, and e (endolysin). The principal results are: (I) All of the genes studied were transcribed exlusively from the so-called l-strand of phage DNA. (2) DNA synthesis and the maturation-defective proteins were required to turn-off transcription of genes 42, rIIA, tIIB, and 43. Experiments performed with chloramphenicol suggested that all phage-specific proteins required to turn-off transcription of these genes were not present until 6 to 8 min post infection (p.i.). (3) During a normal developmental programme, gene 30 was transcribed throughout the eclipse. DNA-negative and maturation-defective conditions had no obvious effect on transcription of this gene. (4) During a normal lytic event, two discrete waves of gene e transcription were observed. The late wave was dependent upon DNA-synthesis and presence of functional maturation-defective proteins. The early wave was unaffected by DNA-negative or maturation-defective conditions. Experiments with chloramphenicol indicated that, if any virus-specific proteins are involved with regulation of early e transcription, such proteins are present by 3 min p.i. The data are interpreted to mean that early gene transcription is regulated by a minimum of two mechanisms. One of these mechanisms is fully operational by the 3rd min and, among the genes studied, controlled early e transcription. A second mechanism becomes operational between 6 and 8 min p.i. and controls transcription of genes 42, 43, rIIA, and rIIB.


LC-MS3 quantification of O-glycopeptides in human serum.
Quantitative analysis of site-specific glycosylation of proteins is a challenging part of glycoproteomic research. Multiple enrichment steps are typically required in the analytical workflows to achieve adequate characterization of the site-specific glycoforms. In spite of recent advances, quantitative workflows need further development. Here, we report a selective and sensitive MS2 followed by further fragmentation in the linear IT-MS analyzer (MS3) multiple reaction monitoring workflow mass spectrometric method for direct analysis of O-glycopeptides in difficult matrix such as serum. Method optimization was performed using two serum glycoproteins, hemopexin (HPX) and sex hormone binding globulin. With the optimized MS3 workflow, we were able to analyze major glycoforms of HPX directly in human serum. Quantification of the minor glycoforms of HPX and glycoforms of sex hormone binding globulin required enrichment of the protein because these analytes were below the sensitivity of the 4000 quadrupole ion trap hybrid mass spectrometer in the complex serum background. In conclusion, we present a quantitative method for site-specific analysis of O-glycosylation with general applicability to mucin-type glycoproteins. Our results document reliable application of the optimized MS3 multiple reaction monitoring workflow to the relative quantification of O-glycosylation microheterogeneity of HPX in human serum. Introduction of isotopically labeled standards would be desirable to achieve absolute quantification of the analytes. The possibility to analyze serum samples directly represents a significant improvement of the quantitative glycopeptide workflows with the potential for use in clinical applications.


The impact of obesity on breast cancer: a retrospective review.
Obesity has been linked to many adverse health consequences, including breast cancer; however, the impact on clinical presentation, tumor characteristics, and survival outcomes has yet to be clearly defined. Retrospective review of a prospectively collected database of patients treated at a single institution for invasive breast cancer from 2000-2008 comparing two groups: nonobese (body mass index of <30) and obese (body mass index of ≥ 30) patients. Continuous variables, categorical variables, and survival data were analyzed. Of 1352 total patients, 76% were classified as nonobese and 24% were obese. When comparing age, obese patients presented less frequently than nonobese patients <50 years old (10% vs. 90%), and when comparing patients >50 years old (18% vs. 82%, P = 0.0019). Obese patients were more likely to present with disease detected by imaging when compared to nonobese patients (67% vs. 56%, P = 0.0006). Obese patients had larger tumors (1.7 cm vs. 1.4 cm, P < 0.001) and higher rates of lymph node (LN) metastases (31% vs. 25%, P = 0.026). On multivariate analysis, obesity was associated with nonpalpable tumors, larger tumors, a higher incidence of LN metastasis, lower incidence of Her2 positivity, lower incidence of multifocality, and less likely to undergo reconstruction after mastectomy. Obese patients clinically present at older ages with mammographically detected breast cancer at more advanced stages than nonobese patients. Strategies to encourage screening among the obese patient population are important.


Evolutionary modifications of human milk composition: evidence from long-chain polyunsaturated fatty acid composition of anthropoid milks.
Brain growth in mammals is associated with increased accretion of long-chain polyunsaturated fatty acids (LCPUFA) in brain phospholipids. The period of maximum accumulation is during the brain growth spurt. Humans have a perinatal brain growth spurt, selectively accumulating docosahexaenoic acid (DHA) and other LCPUFA from the third trimester through the second year of life. The emphasis on rapid postnatal brain growth and LCPUFA transfer during lactation has led to the suggestion that human milk LCPUFA composition may be unique. Our study tests this hypothesis by determining fatty acid composition for 11 species of captive anthropoids (n=53; Callithrix jacchus, Cebus apella, Gorilla gorilla, Hylobates lar, Leontopithecus rosalia, Macaca mulatta, Pan troglodytes, Pan paniscus, Pongo pygmaeus, Saimiri boliviensis, and Symphalangus syndactylus). Results are compared to previously published data on five species of wild anthropoids (n=28; Alouatta paliatta, Callithrix jacchus, Gorilla beringei, Leontopithecus rosalia, and Macaca sinica) and human milk fatty acid profiles. Milk LCPUFA profiles of captive anthropoids (consuming diets with a preformed source of DHA) are similar to milk from women on a Western diet, and those of wild anthropoids are similar to milk from vegan women. Collectively, the range of DHA percent composition values from nonhuman anthropoid milks (0.03-1.1) is nearly identical to that from a cross-cultural analysis of human milk (0.06-1.4). Humans do not appear to be unique in their ability to secrete LCPUFA in milk but may be unique in their access to dietary LCPUFA.


Topotecan given as a 21-day infusion in the treatment of advanced ovarian cancer.
A phase II programme was carried out in both Europe and North America to evaluate the activity of topotecan administered as a 21-day continuous intravenous infusion to patients with recurrent ovarian cancer. The European results are reported here. Patients who had failed first line therapy with a platinum-based regimen received topotecan 0.4 mg/m(2)/day, as a 21-day infusion every 28 days. Patients were only permitted one prior regimen. 35 patients were enrolled and evaluable for response. 3 patients (8.6%) had a partial response to treatment (95% CI 1.8%, 23.1%) with a median time to response of 8.1 weeks and a median duration of response of 17.6 weeks. Response was also evaluated by CA125 and was also found to be 8%. For all 35 patients, median time to progression was 16.1 weeks and median survival was 43.6 weeks. The principal toxicity was myelosuppression although grade 4 neutropenia occurred in only 8.8% of patients (2.1% of courses) and infectious complications were relatively infrequent. Non-haematological toxicity was generally mild and mainly consisted of gastrointestinal events, alopecia and fatigue. A prolonged infusion of topotecan was well tolerated with a low incidence of severe neutropenia. Responses were seen in both North American and European patients. Response rates varied between the 2 studies possibly due to differences in patient demographics.


Axon guidance by gradients of a target-derived component.
Spatial gradients of axon guiding molecules have long been suspected to provide positional and directional cues for retinal ganglion cell axons growing within the optic tectum. With the identification of a guiding activity from tectal cell membranes, it has become possible to investigate the potential physiological significance of molecular gradients for retinal growth cone behavior in vitro. A subset of retinal growth cones, those from the temporal half, were highly sensitive to small concentration changes of the guiding component. The degree of response was correlated with the strength of the gradient. These findings demonstrate that the neural growth cone can read gradients of surface-associated information.


Polyamine metabolism of rat gastric mucosa after oral administration of hypertonic sodium chloride solution.
Oral administration of 1 ml of 3.42 M NaCl solution to rats induced spermidine/spermine N1-acetyltransferase (SSAT) activity in gastric mucosa as well as ornithine decarboxylase (ODC) activity. SSAT activity increased and peaked at 5 h and again at 7 h, whereas ODC activity peaked at 6 h. SSAT mRNA also increased after 3.42 M NaCl administration to an extent similar to the increase in SSAT activity at 5 h. Intracellular putrescine level and DNA synthesis were increased by NaCl administration. A polyamine oxidase inhibitor, N,N'-bis(2,3-butadienyl)-1,4-butanediamine (MDL-72527), but not an ODC inhibitor, alpha-difluoromethylornithine (DFMO), inhibited the increases in putrescine level and DNA synthesis at 5 h. The inhibition of DNA synthesis by MDL-72527 was reversed by putrescine administration. In contrast, both MDL-72527 and DFMO inhibited the increase in putrescine level and DNA synthesis at 16.5 h. These findings suggest that putrescine produced from preexistent spermidine by SSAT is responsible for the initial DNA synthesis after mucosal injury induced by NaCl and that both SSAT and ODC are involved in formation of putrescine, which is required for subsequent DNA synthesis.


Reconstitution of native-like nucleosome core particles from reversed-phase-HPLC-fractionated histones.
We have reconstituted nucleosome core particles from reversed-phase-HPLC-purified chicken erythrocyte core histones and 145 bp random-sequence DNA fragments. Characterization of the resulting nucleoprotein complexes by sedimentation velocity, CD and DNase I footprinting showed that they are structurally indistinguishable from native nucleosome core particles. Furthermore, we have shown that the ability to reproduce these native-like structural features in these reconstituted nucleosome core particles is basically independent of the biological source or the method used (i.e. salt versus acid) for the extraction of histones before their HPLC fractionation. The usefulness and relevance of this approach for the reconstitution of native-like chromatin structures from histone types (histone variants/post-translationally modified histones), which are usually available only in relatively small amounts, is discussed.


[Dialectic hermeneutics: a way of thinking in nursing research].
This paper is a theoretical study about dialectic and hermeneutics union and the possibility of its application as a "way of thinking" in nursing research. For this, we retake its origin and discussion at the present time, as well as the theoretical postulates that gives it foundation, looking forward on expanding the horizons in nursing investigation.


[Use of disodium pamidronate in patients with bone metastases in patients with pulmonary carcinoma].
Disodium pamidronate (Aredia), a drug of the bisphosphonate group, was administered to 20 patients with pains in the bones due to secondary deposits of lung cancer. The aim of the study was to investigate the analgesic effect of pamidronate to osteolytic metastases of lung cancer in the bone. In 16 (80%) patients non-small-cell lung cancer was diagnosed, and in 4 (20%) patients small-cell-lung cancer was confirmed. Intensive disseminated pains in the bones were present in all the patients. Metastases in the skeleton were confirmed by radiography and scintigraphy of the bones. Initial values of calcium in the plasma were determined in those patients. In 11 (55%) patients, initial values of calcium in the serum were normal, and in 9 (45%) patients, they were elevated. Patients with normal calcium values received 30 mg of pamidronate in 250 ml of normal saline, and patients with hypercalcemia received 45-60 mg in 500 ml of normal saline. Analgesic effect of pamidronate was present in 12 (60%) patients, and the completely painless state was achieved in 4 out of 12 patients. Evaluation of the pain was done by questionnaire, using a simple, descriptive scale. In all 9 (100%) patients with hypercalcemia, values of calcium in plasma were normalized. Pamidronate exerted favorable analgesic effect in the case of metastases of lung cancer in the bones in more than 50% patients. Satisfactory results were also achieved in the patients non-responsive to palliative therapy by irradiation. This enables the use of opiates in lower doses.


Pulverulent nuclear cataract in the Norwegian buhund.
Pulverulent cataracts were diagnosed in 52 of 102 Norwegian buhunds, with both sexes being equally affected. All the dogs were otherwise considered clinically healthy. Initial lens changes were visible from six and a half weeks of age as small dots parallel to the suture lines behind the nucleus. Gradually, the opacities along the suture lines became more opaque until, by the age of four to five and a half years, they had progressed to involve the fetal nucleus which then resembled a ball of candy floss. The outer part, the adult nucleus, usually remained clear, and the cortex was not involved. The ages of the dogs in the present study ranged from three and a half weeks to 12 years at first examination. An outcross of an affected bitch to a mixed-breed dog resulted in three puppies, of which two were affected. The retinas could be evaluated in all the examined dogs, and revealed no abnormalities except for one case of focal retinopathy. In addition to the pulverulent cataracts, 10 cases of cortical cataracts were diagnosed, three of which also had pulverulent cataracts. Based on the pedigrees, an autosomal dominant mode of inheritance with a high degree of penetrance is suggested for pulverulent cataract.


The Effect of Organic Acid, Trisodium Phosphate and Essential Oil Component Immersion Treatments on the Microbiology of Cod (Gadus morhua) during Chilled Storage.
Spoilage is a major issue for the seafood sector with the sale and exportation of fish limited by their short shelf-life. The immediate and storage effects of immersion (30 s at 20 °C) with 5% (w/v) citric acid (CA), 5% (v/v) lactic acid (LA), 5% (w/v) capric acid (CP) and 12% trisodium phosphate (TSP) (experiment 1) and essential oil components (EOC) (1% (v/v) citral (CIT), 1% (v/v) carvacrol (CAR), 1% (w/v) thymol (THY) and 1% (v/v) eugenol (EUG)) (experiment 2) on the concentrations of indicator (total viable counts (TVC) (mesophilic and psychrophilic) and total Enterobacteriaceae counts (TEC)), and spoilage organisms (Pseudomonas spp., lactic acid bacteria (LAB), Brochothrix thermosphacta, Photobacterium spp. and hydrogen sulphide producing bacteria (HSPB)) on cod (Gadus morhua) (stored aerobically at 2 °C) was investigated. There was no significant reduction for most treatment-bacteria combinations, with the following exceptions; TSP and TVCm (time t = 6), TSP and TVCp (t = 6), CP and LAB (t = 6, 8 and 10), CP and Br. thermosphacta (t = 4, 6, 8, 10, 14 and 16), TSP and Photobacterium spp. (t = 4), CAR and Br. thermosphacta (t = 6) and CAR and HSPB (t = 3, 6, 9, 12, 15 and 18). Although the majority of treatments did not significantly (P > 0.05) reduce bacterial counts, the limited success with CP and CAR warrants further investigation.


Permeability of Plant Young Root Endodermis to Cu Ions and Cu-Citrate Complexes in Corn and Soybean.
The non-selective apoplastic passage of Cu and Cu-citrate complexes into the root stele of monocotyledonous corn and dicotyledonous soybean was investigated using an inorganic-salt-precipitation technique. Either Cu ions or Cu-citrate complexes were drawn into root through the apoplast from the root growth medium, and K4[Fe(CN)6] was subsequently perfused through xylem vessels or the entire root cross section. Based on microscopic identification of the reddish-brown precipitates of copper ferrocyanide in the cell walls of the xylem of corn and soybean roots, Cu(2+) passed through the endodermal barrier into the xylem of both species. When the solution containing 200 μM CuSO4 and 400 μM sodium citrate (containing 199.98 μM Cu-citrate, 0.02 μM Cu(2+)) was drawn via differential pressure gradients into the root xylem while being perfused with K4[Fe(CN)6] through the entire root cross-section, reddish-brown precipitates were observed in the walls of the stele of soybean, but not corn root. However, when a CuSO4 solution containing 0.02 or 0.2 μM free Cu(2+) was used, no reddish-brown precipitates were detected in the stele of either of the two plants. Results indicated that endodermis was permeable to Cu-citrate complexes in primary roots of soybean, but not corn. The permeability of the endodermal barrier to the Cu-citrate complex may vary between dicotyledonous and monocotyledonous plants, which has considerable implications for chelant-enhanced phytoextraction.


Analysing cluster randomised controlled trials using GLMM, GEE1, GEE2, and QIF: results from four case studies.
Using four case studies, we aim to provide practical guidance and recommendations for the analysis of cluster randomised controlled trials. Four modelling approaches (Generalized Linear Mixed Models with parameters estimated by maximum likelihood/restricted maximum likelihood; Generalized Linear Models with parameters estimated by Generalized Estimating Equations (1st order or second order) and Quadratic Inference Function, for analysing correlated individual participant level outcomes in cluster randomised controlled trials were identified after we reviewed the literature. We systematically searched the online bibliography databases of MEDLINE, EMBASE, PsycINFO (via OVID), CINAHL (via EBSCO), and SCOPUS. We identified the above-mentioned four statistical analytical approaches and applied them to four case studies of cluster randomised controlled trials with the number of clusters ranging from 10 to 100, and individual participants ranging from 748 to 9,207. Results were obtained for both continuous and binary outcomes using R and SAS statistical packages. The intracluster correlation coefficient (ICC) estimates for the case studies were less than 0.05 and are consistent with the observed ICC values commonly reported in primary care and community-based cluster randomised controlled trials. In most cases, the four methods produced similar results. However, in a few analyses, quadratic inference function produced different results compared to the generalized linear mixed model, first-order generalized estimating equations, and second-order generalized estimating equations, especially in trials with small to moderate numbers of clusters. This paper demonstrates the analysis of cluster randomised controlled trials with four modelling approaches. The results obtained were similar in most cases, however, for trials with few clusters we do recommend that the quadratic inference function should be used with caution, and where possible a small sample correction should be used. The generalisability of our results is limited to studies with similar features to our case studies, for example, studies with a similar-sized ICC. It is important to conduct simulation studies to comprehensively evaluate the performance of the four modelling approaches.


Unveiling the link between inflammasomes and skin cutaneous melanoma: Insights into expression patterns and immunotherapy response prediction.
Skin cutaneous melanoma (SKCM) is one of the most malignant forms of skin cancer, characterized by its high metastatic potential and low cure rate in advanced stages. Despite advancements in clinical therapies, the overall cure rate for SKCM remains low due to its resistance to conventional treatments. Inflammation is associated with the activation and regulation of inflammatory responses and plays a crucial role in the immune system. It has been implicated in various physiological and pathological processes, including cancer. However, the mechanisms of inflammasome activation in SKCM remain largely unexplored. In this study, we quantified the expression level of six inflammasome-related gene sets using transcriptomic data from SKCM patients. As a result, we found that inflammasome features were closely associated with various clinical characteristics and served as a favorable prognostic factor for patients. A functional enrichment analysis revealed the oncogenic role of inflammasome features in SKCM. Unsupervised clustering was applied to identify immune clusters and inflammatory subtypes, revealing a significant overlap between immune cluster 4 and SKCM subtype 2. The CASP1, GSDMD, NLRP3, IL1B, and IL18 features could predict immune checkpoint blockade therapy response in various SKCM cohorts. In conclusion, our study highlighted the significant association between the inflammasome and cancer treatment. Understanding the role of inflammasome signaling in SKCM pathology can help identify potential therapeutic targets and improve patient prognosis.


Experimental application of Business Process Management technology to manage clinical pathways: a pediatric kidney transplantation follow up case.
To test the application of Business Process Management technology to manage clinical pathways, using a pediatric kidney transplantation as case study, and to identify the benefits obtained from using this technology. Using a Business Process Management platform, we implemented a specific application to manage the clinical pathway of pediatric patients, and monitored the activities of the coordinator in charge of the case management during a 6-month period (from June 2015 to November 2015) using two methodologies: the traditional procedure and the one under study. The application helped physicians and nurses to optimize the amount of time and resources devoted to management purposes. In particular, time reduction was close to 60%. In addition, the reduction of data duplication, the integrated event management and the efficient data collection improved the quality of the service. The use of Business Process Management technology, usually related to well-defined processes with high management costs, is an established procedure in multiple environments; its use in healthcare, however, is innovative. The use of already accepted clinical pathways is known to improve outcomes. The combination of these two techniques, well established in their respective areas of application, could represent a revolution in clinical pathway management. The study has demonstrated that the use of this technology in a clinical environment, using a proper architecture and identifying a well-defined process, leads to real benefits in terms of resources optimization and quality improvement.


How I overcame problems in in vitro fertilisation of livestock animals.
In my research life of 35years, growing with IETS as a researcher of in vitro maturation and fertilisation (IVM/IVF) of porcine and cattle oocytes, I suffered from hard times related to solving problems that prevented the progress of my research and conferment of my degrees. Many researchers may have similar problems. Thus, I would like to address a few examples of how I overcame these problems related to IVF and help young researchers with similar troubles. There were four main problems to be solved in my experiments. Problem 1: Establishment of IVF using only defined medium in pigs. Problem 2: Establishment of successful in vitro culture (IVC) of IVM/IVF bovine oocytes in defined medium. Problem 3: Low rate of male pronucleus formation in IVM porcine oocytes after IVF. Problem 4: Sedimentation of Ca2+ in the sperm capacitation solution for IVF in pigs. Problem 1 was solved by a lucky accident, in which a sperm suspension that would have otherwise been discarded happened to be successfully used for IVF in pigs. Problems 2, 3 and 4 were solved by communication with scientists whose fields were different from mine, where similar problems had been solved already. Young researchers are encouraged to transcend the boundaries of their research fields and solve problems by interacting with researchers in different fields. There are many good connections or answers around us that may be effective in resolving the problems that are hindering the progress of pending research.


Hippocampal metabolite abnormalities in fibromyalgia: correlation with clinical features.
Although the pathology of fibromyalgia is poorly understood, a growing body of evidence suggests involvement of the central nervous system. The hippocampus is a brain center that is sensitive to the effects of stress exposure and has been demonstrated to be affected in a variety of disorders whose onset, like fibromyalgia, are associated with stressful experience. We therefore interrogated the bilateral hippocampus of 16 female fibromyalgia patients in comparison to 8 age- and gender-matched healthy control subjects using single voxel proton magnetic resonance spectroscopy. Our results demonstrate a significant reduction in the ratio of N-acetylaspartate to creatine (NAA/Cr) in fibromyalgia patients versus matched control subjects specifically in the right temporal lobe from a voxel centered on the right hippocampus (patient vs control, mean +/- standard deviation: 1.20 +/- 0.13 vs 1.34 +/- 0.10, P = .03). Moreover, correlation analysis demonstrated a significant negative correlation between patient scores on the Fibromyalgia Impact Questionnaire and NAA/Cr ratio within the right hippocampus (Spearman rank correlation, rho = -0.681, P = .018). Our results indicate that fibromyalgia is associated with brain metabolite abnormalities within the right hippocampus that correlate with patient symptoms. We have demonstrated an abnormality in hippocampal brain metabolites in premenopausal female fibromyalgia patients with no psychiatric comorbidity. A significant negative correlation between patient subjective experience of symptoms and a reduced NAA/Cr ratio suggests a role for hippocampal pathology in fibromyalgia.


Evaluation of head and neck squamous cell carcinoma invasiveness by the electrical resistance breakdown assay.
Invasion of tumor cells into the surrounding tissue is a hallmark of cancer. Squamous cell carcinomas of the head and neck region (HNSCC) are characterized by their early primarily lymphatic metastatic spread. The aim of the present study was to evaluate the use of the electrical resistance breakdown assay for determining HNSCC tumor cell invasiveness. The assay utilizes the high transepithelial electrical resistance (TEER) of an epithelial MDCK-C7 monolayer as a sensitive indicator of monolayer integrity and permeability. MDCK-C7 cells were grown to confluence in microfilter membrane cups. 3 x 10(6) cancer cells of cell lines UM-SCC-3, UM-SCC-27, UMB-SCC-745, UMB-SCC-864, UMB-SCC-969 and UT-SCC-26A derived from HNSCC tumors, were seeded on top of this epithelial test barrier. A7-melanoma cells served as a positive control whereas MDCK-C7 cells were used as a negative control and were applied in the same number as the tested tumor cells. TEER was measured over the following days and compared to control values. A significant reduction in TEER was observed in the UMB-SCC-745, UMB-SCC-969 and UT-SCC-26A cell lines within the first 72 h, whereas no significant reduction in TEER was seen in the UM-SCC-3, UM-SCC-27 and UMB-SCC-864 cell lines. HNSCC cell lines in general are found to be less invasive in the resistance breakdown assay compared to other tumor cells such as A7-melanoma cells, however, the electrical resistance breakdown assay appears capable of demonstrating differences in invasiveness between different HNSCC cell lines and therefore potentially could serve as a versatile tool in distinguishing high and low invasive tumors with a potential application as a diagnostic and prognostic marker in clinical investigations.


[The diagnosis artifice].
The clinical diagnosis is considered the cornerstone in the decisions making process, since it has various functions, including the executive. However, the diagnosis is not only a goal by itself, but a mean to help patients. It is not enough to identify how the disease which ails the patient is called; the diagnosis must be individualized as much as possible in each case, for the mere mention of the diagnosis can mean a wide variety of problems.


Pharmaceutical Interventions for Diabetes Prevention in Patients at Risk.
With the rising incidence and prevalence rates of type 2 diabetes globally, it is imperative that diabetes prevention strategies are implemented to stem the flow of new cases. Successful interventions include both lifestyle modification and pharmaceutical agents, and large, multicentre, randomised, controlled studies in different populations have identified the benefits of both. However, translating positive trial outcomes to the real world is particularly challenging, as lifestyle interventions require regular reinforcement from healthcare professionals to be maintained. Pharmaceutical therapies may therefore play an adjunctive role in combination with lifestyle to prevent diabetes. Population-based strategies are also necessary to reduce sedentary behaviour and obesity. Well-established glucose-lowering therapies such as metformin, sulphonylureas, thiazolidinediones and insulin and newer agents such as incretin therapies and sodium glucose co-transporter 2 inhibitors have all been investigated in randomised controlled trials for diabetes prevention with varying success. Non-glucose-lowering therapies such as orlistat and renin angiotensin system blockers can prevent diabetes, whereas statins are associated with slightly increased risk. Diabetes prevention strategies should carefully consider the use of these agents according to individual patient circumstances and phenotypic profile.


The avian haemophili.
There are four currently recognized taxa to accommodate the avian haemophili: Haemophilus paragallinarum, Pasteurella avium, Pasteurella volantium, and Pasteurella species A (the last three being formerly united as Haemophilus avium). A range of other taxa has also been recognized, but they have been neither named nor assigned to a genus. All of these various taxa, legitimate and otherwise, have the common characteristic of requiring V factor, but not X factor, for in vitro growth. Several recent studies have established the phenotypic properties that allow the differentiation of the recognized taxa, both named and unnamed. The serological properties of H. paragallinarum, the causative agent of infectious coryza of chickens, has received considerable recent attention. In contrast, many questions on the pathogenicity and virulence mechanisms of H. paragallinarum remain unanswered. Another area requiring further work is the identification of those antigens responsible for inducing protective immunity in vaccinated or naturally infected chickens.


Genomic architecture constrains macromolecular allocation in dinoflagellates.
Dinoflagellate genomes have a unique architecture that may constrain their physiological and biochemical responsiveness to environmental stressors. Here we quantified how nitrogen (N) starvation influenced macromolecular allocation and C:N:P of three photosynthetic marine dinoflagellates, representing different taxonomic classes and genome sizes. Dinoflagellates respond to nitrogen starvation by decreasing cellular nitrogen, protein and RNA content, but unlike many other eukaryotic phytoplankton examined RNA:protein is invariant. Additionally, 2 of the 3 species exhibit increases in cellular phosphorus and very little change in cellular carbon with N-starvation. As a consequence, N starvation induces moderate increases in C:N, but extreme decreases in N:P and C:P, relative to diatoms. Dinoflagellate DNA content relative to total C, N and P is much higher than similar sized diatoms, but similar to very small photosynthetic picoeukaryotes such as Ostreococcus. In aggregate these results indicate the accumulation of phosphate stores may be an important strategy employed by dinoflagellates to meet P requirements associated with the maintenance and replication of their large genomes.


Assessing protein immunogenicity with a dendritic cell line-derived endolysosomal degradome.
The growing number of novel candidate molecules for the treatment of allergic diseases imposed a dramatic increase in the demand for animal experiments to select immunogenic vaccines, a pre-requisite for efficacy. Because no in vitro methods to predict the immunogenicity of a protein are currently available, we developed an in vitro assay that exploits the link between a protein's immunogenicity and its susceptibility to endolysosomal proteolysis. We compared protein composition and proteolytic activity of endolysosomal fractions isolated from murine bone marrow- and human blood- derived dendritic cells, and from the dendritic cell line JAWS II. Three groups of structurally related antigen variants differing in their ability to elicit immune responses in vivo (Bet v 1.0101 and Bet v 1.0401, RNases A and S, holo- and apo-HRP) were subjected to in vitro simulated endolysosomal degradation. Kinetics and patterns of generated proteolytic peptides were evaluated by gel electrophoresis and mass spectrometry. Antigens displaying weak capacity of T cell priming in vivo were highly susceptible to endolysosomal proteases in vitro. As proteolytic composition, activity, and specificity of endolysosomal fractions derived from human and murine dendritic cells were comparable, the JAWS II cell line could be used as a substitute for freshly isolated human or murine cells in in vitro degradation assays. Endolysosomal fractions prepared from the JAWS II cell line provide a reliable tool for in vitro estimation of protein immunogenicity. The rapid and simple assay described here is very useful to study the immunogenic properties of a protein, and can help to replace, reduce, and refine animal experiments in allergy research and vaccine development in general.


The relation of etiology based on the 2017 ILAE classification to the effectiveness of the ketogenic diet in drug-resistant epilepsy in childhood.
To investigate the effectiveness and safety of the ketogenic diet (KD) in drug-resistant epilepsy in childhood in relation to the new 2017 International League Against Epilepsy (ILAE) classification of etiology. A consecutive cohort of patients treated with the KD were categorized according to the ILAE classification into known (structural, genetic, metabolic, infectious, and immune-mediated) and unknown etiology. Primary outcome was the frequency of patients achieving seizure freedom with the KD at 3 months, secondary outcomes were seizure reduction >50% at 3 months, and both seizure freedom and seizure reduction >50% at 6, 12 months, and at last follow-up (LFU), and adverse effects. Outcomes were compared between etiology groups. Etiology was known in 70% (129/183). Outcomes did not differ at 3 months (known vs unknown: seizure freedom 28% vs 33%, seizure reduction 62 vs 67%), but seizure freedom was significantly less frequent in known etiology at 6 months (26% vs 43%) and beyond (22% vs 37%). Logistic regression identified duration of epilepsy, number of previous antiseizure medications (ASMs), and age-appropriate psychomotor development as positive determinants of outcome. Among individual etiology groups, the effectiveness of KD was relatively best for genetic (33% at LFU) and poorest for metabolic etiology (8% at LFU). The small number of patients with infectious and immune-mediated etiology requires larger numbers in each etiology group to corroborate our results. No differences in type and frequency of adverse effects (in 71%) between etiology groups were observed, requiring medical intervention in 21%. The KD was most effective in genetic and unknown etiology, many unknowns probably represent yet unidentified genetic causes. We recommend consequent diagnostic and genetic work-up to identify etiologies that respond best to the KD. The KD should be offered early to infants with genetic epilepsy before deterioration of epileptic symptoms and of psychomotor development.


A mouse model of chronic bacterial lesions (a cotton trap) for studying oral bacteria-lymphocyte interactions.
We established a mouse model of chronic bacterial infection (cotton trap) to get a deeper insight into interactions between immune cells and bacterial strains, that are most commonly isolated from periapical processes. We have used flow cytometry to identify the presence of intracellular cytokines of activated T cells collected from cotton traps, previously infected with different strains of bacteria and implanted subcutaneously into the back of the mice. We provide an evidence that anaerobic bacteria (Bacteroides sp.) and nocardiae are more effective in inducing cytotoxic immunity and Th1 response compared to oral streptococci. Differences in immune response against anaerobic bacteria when compared to streptococci are probably dependent on some non-specific immune cell stimulation (e.g. by bacterial cell wall components), nevertheless the role of specific antigen-dependent immune mechanism can not be excluded.


[Assessment of the separate elements of an epidemiological overview of poliomyelitis today].
The materials on the epidemiological supervision of poliomyelitis in a large city, carried out in 1972-1981, are presented. Two periods differing in morbidity rate, in the level of population immunity to poliomyelitis and in the activity of the poliovirus circulation are defined. The supervision of poliomyelitis is shown to be capable of influencing the course of the epidemic process of this infection provided that the individual elements of such supervision are correctly evaluated in different epidemic situations.


Hearing, single-unit analysis, and vocalizations in songbirds.
Auditory neurons in the avian cochlear nucleus are systematically arranged according to their best frequencies. The thresholds of single auditory neurons at their best frequencies match the thresholds of hearing obtained by behavioral methods for the same frequencies. The upper range of single-unit best frequencies shows distinct interspecific variation which is correlated with the differences in the range of vocal frequencies. Most songbirds do not seem to produce frequencies below I kilohertz but can hear them.


Decolorization of molasses fermentation wastewater by SnO(2)-catalyzed ozonation.
In the presence of O(3), the oxidative decolorization reaction on molasses fermentation wastewater with SnO(2) as a catalyst was studied. The results showed that SnO(2) accelerated the ozone oxidation reaction and the oxidative decolorization of molasses fermentation wastewater was accelerated. Influences on SnO(2) catalytic ozonation activity by precipitants and the calcination temperature were studied by XRD, IR and TG-DSC. SnO(2) prepared by ammonia as the precipitant had higher catalytic activity and a stronger dehydroxylation. The IR spectra of adsorbed pyridine showed that there were Lewis acid sites on the surface of this SnO(2) catalyst. The main factors influencing molasses fermentation wastewater oxidative decolorization were the wastewater concentration, the O(3) concentration, the pH value and the catalyst dosage. The decolorization of wastewater was improved with the increase of the wastewater dilution ratio, the ozone concentration and the catalyst dosage. High activity *OH was found to be existing with less amount and low concentration in the process of SnO(2) catalyzed ozonation decolorization.


Vaccination for the expatriate and long-term traveler.
Duration of travel is an important factor in addressing travel health safety due to cumulative risk of exposure to illness and injury. The diverse group of expatriate and long-term business and leisure travelers present a different spectrum of issues for the travel medicine practitioner to address during consultation than does the short-term traveler, due to changes in travel patterns and activities, lifestyle alterations, and increased interaction with local populations. Immunization provides one safe and reliable method of preventing infectious illness in this group. We review travel patterns and available data on illnesses that they may be exposed to, including the increased risk of certain vaccine-preventable illnesses. We review the pre-travel management of these travelers, particularly the increased risk of certain vaccine-preventable illnesses as it applies to routine vaccines, recommended travel vaccines and required travel vaccines.


The impact of evidence-based practice in nursing and the next big ideas.
The impact of evidence-based practice (EBP) has echoed across nursing practice, education, and science. The call for evidence-based quality improvement and healthcare transformation underscores the need for redesigning care that is effective, safe, and efficient. In line with multiple direction-setting recommendations from national experts, nurses have responded to launch initiatives that maximize the valuable contributions that nurses have made, can make, and will make, to fully deliver on the promise of EBP. Such initiatives include practice adoption; education and curricular realignment; model and theory development; scientific engagement in the new fields of research; and development of a national research network to study improvement. This article briefly describes the EBP movement and considers some of the impact of EBP on nursing practice, models and frameworks, education, and research. The article concludes with discussion of the next big ideas in EBP, based on two federal initiatives, and considers opportunities and challenges as EBP continues to support other exciting new thinking in healthcare.