Glycosylated RGD-containing peptides: tracer for tumor targeting and angiogenesis imaging with improved biokinetics.
The alpha(v)beta3 integrin plays an important role in metastasis and tumor-induced angiogenesis. Targeting with radiolabeled ligands of the alpha(v)beta3 integrin may provide information about the receptor status and enable specific therapeutic planning. Previous studies from our group resulted in tracers that showed alpha(v)beta3-selective tumor uptake. However, these first-generation compounds predominantly revealed hepatobiliary excretion with high radioactivity found in the liver. In this report, the synthesis and biological evaluation of the first glycosylated RGD-containing peptide (RGD-peptide) for the noninvasive imaging of alpha(v)beta3 expression are described. Peptides were assembled on a solid support using fluorenylmethoxycarbonyl-coupling protocols. The precursor cyclo(-Arg-Gly-Asp-D-Tyr-Lys(SAA)-) GP1 was synthesized by coupling 3-acetamido-2,6-anhydro-4,5,7-tri-O-benzyl-3-deoxy-beta-D-glycero-D-gulo-heptonic acid (SAA(Bn3)) with cyclo(-Arg(Mtr)-Gly-Asp(OtBu)-D-Tyr(tBu)-Lys-) and subsequent removal of the protection groups. Iodine labeling was performed by the Iodo-Gen method (radiochemical yield > 50%). The in vitro binding assays were performed using purified immobilized alpha(IIb)beta3, alpha(v)beta5, and alpha(v)beta3 integrins. For in vivo experiments, nude mice bearing xenotransplanted melanomas and mice with osteosarcomas were used. The glycosylated peptide 3-iodo-Tyr4-cyclo(-Arg-Gly-Asp-D-Tyr-Lys(SAA)-) GP2 showed high affinity and selectivity for alpha(v)beta3 in vitro (50% inhibitory concentration = 40 nmol/L). Pretreatment studies indicate specific binding of [125I]GP2 on alpha(v)beta3-expressing tumors in vivo. Comparison of the pharmacokinetics of [125I]GP2 and [125I]-3-iodo-Tyr4-cyclo(-Arg-Gly-Asp-D-Tyr-Val-) [125I]P2 revealed for [125I]GP2 an increased activity concentration in the blood (e.g., 3.59 +/- 0.35 percentage injected dose [%ID]/g vs. 1.72 +/- 0.44 %ID/g at 10 min postinjection) and a significantly reduced uptake in the liver (e.g., 2.59 +/- 0.24 %ID/g vs. 21.96 +/- 2.78 %ID/g at 10 min postinjection). Furthermore, a clearly increased activity accumulation in the tumor was found (e.g., 3.05 +/- 0.31 %ID/g vs. 0.92 +/- 0.16 %ID/g at 240 min postinjection), which remained almost constant between 60 and 240 min postinjection. This resulted in good tumor-to-organ ratios for the glycosylated tracer (e.g., 240-min postinjection osteosarcoma model: tumor-to-blood = 16; tumor-to-muscle = 7; tumor-to-liver = 2.5), which were confirmed by the first gamma-camera images of osteosarcoma-bearing mice at 240 min postinjection. This study demonstrates that the introduction of a sugar moiety improves the pharmakokinetic behavior of a hydrophobic peptide-based tracer. Additionally, this alpha(v)beta3-selective glycosylated radioiodinated second-generation tracer GP2 shows high tumor uptake and good tumor-to-organ ratios that allow noninvasive visualization of alpha(v)beta3-expressing tumors and monitoring therapy with alpha(v)beta3 antagonists. Finally, the favorable biokinetics make the glycosylated RGD-peptide a promising lead structure for tracers to quantify the alpha(v)beta3 expression using PET.


Building consensus on interactions between population health researchers and the food industry: Two-stage, online, international Delphi study and stakeholder survey.
Key to scientific integrity is ensuring that research findings are considered credible by scientific peers, practitioners, policymakers and the public. Industry sponsorship of nutritional research can result in bias and raises significant professional, public and media concern. Yet, there is no international consensus on how to prevent or manage conflicts of interest for researchers considering engaging with the food industry. This study aimed to determine internationally agreed principles to guide interactions between population health researchers and the food industry to prevent or manage conflicts of interest. We used a two-stage, online Delphi study for researchers (n = 100 in 28 countries), and an online survey for stakeholders (n = 84 in 26 countries). Levels of agreement were sought with 56 principles derived from a systematic review. Respondent comments were analysed using qualitative content analysis. High levels of agreement on principles were achieved for both groups (researchers 68%; stakeholders 65%). Highest levels of agreement were with principles concerning research methods and governance. More contentious were principles that required values-based decision-making, such as determining which elements of the commercial sector are acceptable to interact with. These results provide the basis for developing internationally-agreed guidelines for population health researchers governing interactions with the food industry.


Lipoprotein concentrations over time in the intensive care unit COVID-19 patients: Results from the ApoCOVID study.
Severe acute respiratory syndrome coronavirus2 has caused a global pandemic of coronavirus disease 2019 (COVID-19). High-density lipoproteins (HDLs), particles chiefly known for their reverse cholesterol transport function, also display pleiotropic properties, including anti-inflammatory or antioxidant functions. HDLs and low-density lipoproteins (LDLs) can neutralize lipopolysaccharides and increase bacterial clearance. HDL cholesterol (HDL-C) and LDL cholesterol (LDL-C) decrease during bacterial sepsis, and an association has been reported between low lipoprotein levels and poor patient outcomes. The goal of this study was to characterize the lipoprotein profiles of severe ICU patients hospitalized for COVID-19 pneumonia and to assess their changes during bacterial ventilator-associated pneumonia (VAP) superinfection. A prospective study was conducted in a university hospital ICU. All consecutive patients admitted for COVID-19 pneumonia were included. Lipoprotein levels were assessed at admission and daily thereafter. The assessed outcomes were survival at 28 days and the incidence of VAP. A total of 48 patients were included. Upon admission, lipoprotein concentrations were low, typically under the reference values ([HDL-C] = 0.7[0.5-0.9] mmol/L; [LDL-C] = 1.8[1.3-2.3] mmol/L). A statistically significant increase in HDL-C and LDL-C over time during the ICU stay was found. There was no relationship between HDL-C and LDL-C concentrations and mortality on day 28 (log-rank p = 0.554 and p = 0.083, respectively). A comparison of alive and dead patients on day 28 did not reveal any differences in HDL-C and LDL-C concentrations over time. Bacterial VAP was frequent (64%). An association was observed between HDL-C and LDL-C concentrations on the day of the first VAP diagnosis and mortality ([HDL-C] = 0.6[0.5-0.9] mmol/L in survivors vs. [HDL-C] = 0.5[0.3-0.6] mmol/L in nonsurvivors, p = 0.036; [LDL-C] = 2.2[1.9-3.0] mmol/L in survivors vs. [LDL-C] = 1.3[0.9-2.0] mmol/L in nonsurvivors, p = 0.006). HDL-C and LDL-C concentrations upon ICU admission are low in severe COVID-19 pneumonia patients but are not associated with poor outcomes. However, low lipoprotein concentrations in the case of bacterial superinfection during ICU hospitalization are associated with mortality, which reinforces the potential role of these particles during bacterial sepsis.


Bryostatin 1 acts synergistically with interleukin-1 alpha to induce secretion of G-CSF and other cytokines from marrow stromal cells.
The protein kinase C (PKC) activator bryostatin 1 (bryo) has substantial antileukemic and hematopoietic actions. Bryo promotes the in vitro growth of normal hematopoietic progenitors by inducing the release of growth factors from accessory cells. We have examined the effects of bryo on the expression and release of certain myeloid growth factors from fibroblastlike marrow stromal cells (MSC). Substantial release of granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF). or interleukin-6 (IL-6) following bryo treatment was seen only in MSC cultures contaminated with macrophages. Bryo alone was ineffective in inducing release of the cytokines from MSC cultures containing only fibroblastlike stromal cells. When MSC were treated with IL-1alpha, substantial quantities of the cytokines (G-CSF, GM-CSF,IL-6) were released. Bryo acted synergistically with IL-1 alpha to significantly increase cytokine release to- to nine-fold compared to IL-1alpha alone (p < 0.016). Neither Il-1alpha nor bryo, alone or in combination, induced release of stem cell factor (scf) from MSC. The synergistic interaction between IL-1alpha and bryo was dose- and schedule-dependent, requiring simultaneous application of IL-1alpha and bryo for optimum effect. Bryo alone induced no G-CSF mRNA accumulation but increased the level seen with IL-1alpha treatment by 50%. The synergistic interaction of bryo and IL-1alpha required PKC, since it was antagonized by agents which depleted or inhibited PKC but not by a protein kinase A antagonist. The increase in G-CSF mRNA was associated with a marked increase in mRNA stability. Bryostatin may promote the release of cytokines from several accessory cell populations, including MSC, to accomplish its in vivo hematopoietic effects.


Anthocyanin characterization, total phenolic quantification and antioxidant features of some Chilean edible berry extracts.
The anthocyanin composition and HPLC fingerprints of six small berries endemic of the VIII region of Chile were investigated using high resolution mass analysis for the first time (HR-ToF-ESI-MS). The antioxidant features of the six endemic species were compared, including a variety of blueberries which is one of the most commercially significant berry crops in Chile. The anthocyanin fingerprints obtained for the fruits were compared and correlated with the antioxidant features measured by the bleaching of the DPPH radical, the ferric reducing antioxidant power (FRAP), the superoxide anion scavenging activity assay (SA), and total content of phenolics, flavonoids and anthocyanins measured by spectroscopic methods. Thirty one anthocyanins were identified, and the major ones were quantified by HPLC-DAD, mostly branched 3-O-glycosides of delphinidin, cyanidin, petunidin, peonidin and malvidin. Three phenolic acids (feruloylquinic acid, chlorogenic acid, and neochlorogenic acid) and five flavonols (hyperoside, isoquercitrin, quercetin, rutin, myricetin and isorhamnetin) were also identified. Calafate fruits showed the highest antioxidant activity (2.33 ± 0.21 μg/mL in the DPPH assay), followed by blueberry (3.32 ± 0.18 μg/mL), and arrayán (5.88 ± 0.21), respectively.


The economic burden of myalgic encephalomyelitis/chronic fatigue syndrome in Australia.
Objective This study aimed to estimate costs of myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) to patients, government and Australian society. Methods Australian ME/CFS patients and their carers were recruited using convenience sampling. Patients completed an online retrospective cost diary, providing ME/CFS-related direct medical, non-medical and indirect costs. Informal care costs were collected directly from carers. Data from the Pharmaceutical Benefits Scheme and Medicare Benefits Schedule were linked to participant survey data. Annual per patient and total societal costs were estimated, broken down by category and presented in 2021 AUD. Factors associated with higher costs were investigated using generalised linear models. Results One hundred and seventy five patients (mean age 49 years s.d. 14, 79.4% female) completed the cost diary. Estimated total annual societal costs of ME/CFS in Australia ranged between $1.38 and $10.09 billion, with average annual total costs of $63 400/patient. Three-quarters of these costs were due to indirect costs ($46 731). Disability severity was the key factor associated with higher costs, particularly for indirect costs (being 2.27-fold higher for severe disability than no/mild disability). Conclusions ME/CFS poses a significant economic burden in Australia, owing mainly to high indirect and informal care costs.


Antimicrobial protegrin-1 forms amyloid-like fibrils with rapid kinetics suggesting a functional link.
Protegrin-1 (PG-1) is an 18 residues long, cysteine-rich β-sheet antimicrobial peptide (AMP). PG-1 induces strong cytotoxic activities on cell membrane and acts as a potent antibiotic agent. Earlier we reported that its cytotoxicity is mediated by its channel-forming ability. In this study, we have examined the amyloidogenic fibril formation properties of PG-1 in comparison with a well-defined amyloid, the amyloid-β (Aβ(1-42)) peptide. We have used atomic force microscopy (AFM) and thioflavin-T staining to investigate the kinetics of PG-1 fibrils growth and molecular dynamics simulations to elucidate the underlying mechanism. AFM images of PG-1 on a highly hydrophilic surface (mica) show fibrils with morphological similarities to Aβ(1-42) fibrils. Real-time AFM imaging of fibril growth suggests that PG-1 fibril growth follows a relatively fast kinetics compared to the Aβ(1-42) fibrils. The AFM results are in close agreement with results from thioflavin-T staining data. Furthermore, the results indicate that PG-1 forms fibrils in solution. Significantly, in contrast, we do not detect fibrillar structures of PG-1 on an anionic lipid bilayer 2-dioleoyl-sn-glycero-3-phospho-L-serine/1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine; only small PG-1 oligomers can be observed. Molecular dynamics simulations are able to identify the presence of these small oligomers on the membrane bilayer. Thus, our current results show that cytotoxic AMP PG-1 is amyloidogenic and capable of forming fibrils. Overall, comparing β-rich AMPs and amyloids such as Aβ, in addition to cytotoxicity and amyloidogenicity, they share a common structural motif, and are channel forming. These combined properties support a functional relationship between amyloidogenic peptides and β-sheet-rich cytolytic AMPs, suggesting that amyloids channels may have an antimicrobial function.


Synthesis of poly(acrylic acid) coated-Fe3O4 superparamagnetic nano-composites and their fast removal of dye from aqueous solution.
Poly(acrylic acid) (PAA) coated-Fe3O4 superparamagnetic nano-composites were synthesized through a solvothermal technique by using cheap and environmental friendly iron salts and PAA. Each nano-composite was composed of many small primary nanocrystals. The as-synthesized products were characterized by X-ray diffraction (XRD), field emission scanning electron microscopy (FE-SEM), high resolution transmission electron microscopy (HRTEM), vibrating sample magnetometer (VSM), Fourier transform infrared spectroscopy (FTIR), thermogravimetric-differential scanning calorimetry (TG-DSC) analysis and nitrogen adsorption-desorption measurements. These nano-composites showed rapid adsorption rate and super removal capacity of cation organic dye methylene blue (MB). Moreover, the MB desorption could be easily performed using methanol and acetic acid mixed solution. The reused performance of the nano-composites was also studied.


Chylomicron amyloid-beta in the aetiology of Alzheimer's disease.
Alzheimer's disease is characterized by inflammatory proteinaceous deposits comprised principally of the protein amyloid-beta (Abeta). Presently, the origins of cerebral amyloid deposits are controversial, though pivotal for the prevention of Alzheimer's disease. Recent evidence suggests that in blood, Abeta may serve as a regulating apoprotein of the triglyceride-rich-lipoproteins and we have found that the synthesis of Abeta in enterocytes and thereafter secretion as part of the chylomicron cascade is regulated by dietary fats. It is our contention that chronically elevated plasma levels of Abeta in response to diets rich in saturated fats may lead to disturbances within the cerebrovasculature and exaggerated blood-to-brain delivery of circulating Abeta, thereby exacerbating amyloidosis. Consistent with this hypothesis we show that enterocytic Abeta is increased concomitant with apolipoprotein B48. Furthermore, cerebral extravasation of immunoglobulin G, a surrogate marker of plasma proteins is observed in a murine model of Alzheimer's disease maintained on a saturated-fat diet and there is diminished expression of occludin within the cerebrovasculature, an endothelial tight junction protein.


[Adnexal masses: understanding and management].
Asymptomatic adnexal masses have become common findings due to the widespread use of the transvaginal ultrasound. Most asymptomatic adnexal masses are benign, but malignancy should still be excluded. The assessment of women with adnexal masses, intended to detect the ones suspicious for malignancy, includes the medical history, physical examination, ultrasound scan with Doppler flows, and, in some cases, the CA-125 levels. Once the patient has completed the primary assessment, the important decision is whether to follow her conservatively (if a benign condition is probable), or to proceed to surgery (if malignancy is suspected). This review discusses the evaluation, differential diagnosis, and general approach to adnexal masses, and proposes a management flow chart.


Autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy: report of seven additional sicilian patients and overview of the overall series from sicily.
Autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED) is a rare recessive inherited disease caused by the mutation of the AIRE gene on chromosome 21. To date, 8 Sicilian patients have been described and the R203X AIRE mutation was found to be the most common in this region. (1) To describe 7 additional Sicilian APECED patients and to review all 15 Sicilian APECED patients who have been investigated by our group in the last years, and (2) to report a novel AIRE gene mutation. Among the 3 cardinal features of APECED, hypoparathyroidism has been already detected in all 15 patients, whereas Addison's disease and chronic mucocutaneous candidiasis have so far been found in 10/15 and 12/15 cases, respectively. In 2 consanguineous cases, AIRE gene analysis revealed a novel mutation, named IVS13+2T, in homozygosis. R203X was the most common mutation in this region (30% of alleles and 46.6% of patients), followed by R257X (20% of alleles and 40% of patients). Sicilian APECED patients are confirmed to have some peculiar characteristics from a clinical and genetic point of view. No correlations between genotype and phenotype were identified.


How should the psychological well-being of zoo elephants be objectively investigated?
Animal welfare (sometimes termed "well-being") is about feelings - states such as "suffering" or "contentment" that we can infer but cannot measure directly. Welfare indices have been developed from two main sources: studies of suffering humans, and of research animals deliberately subjected to challenges known to affect emotional state. We briefly review the resulting indices here, and discuss how well they are understood for elephants, since objective welfare assessment should play a central role in evidence-based elephant management. We cover behavioral and cognitive responses (approach/avoidance; intention, redirected and displacement activities; vigilance/startle; warning signals; cognitive biases, apathy and depression-like changes; stereotypic behavior); physiological responses (sympathetic responses; corticosteroid output - often assayed non-invasively via urine, feces or even hair; other aspects of HPA function, e.g. adrenal hypertrophy); and the potential negative effects of prolonged stress on reproduction (e.g. reduced gametogenesis; low libido; elevated still-birth rates; poor maternal care) and health (e.g. poor wound-healing; enhanced disease rates; shortened lifespans). The best validated, most used welfare indices for elephants are corticosteroid outputs and stereotypic behavior. Indices suggested as valid, partially validated, and/or validated but not yet applied within zoos include: measures of preference/avoidance; displacement movements; vocal/postural signals of affective (emotional) state; startle/vigilance; apathy; salivary and urinary epinephrine; female acyclity; infant mortality rates; skin/foot infections; cardio-vascular disease; and premature adult death. Potentially useful indices that have not yet attracted any validation work in elephants include: operant responding and place preference tests; intention and vacuum movements; fear/stress pheromone release; cognitive biases; heart rate, pupil dilation and blood pressure; corticosteroid assay from hair, especially tail-hairs (to access endocrine events up to a year ago); adrenal hypertrophy; male infertility; prolactinemia; and immunological changes.


[Infestation of the human digestive system with beetle larvae (Coleoptera: Cantharidae): a case report].
This study was conducted to report the digestive system infestation caused by the larvae of Coleoptera in a female pediatric patient. She was admitted to our hospital with the complaints of emergence of insect larvae from her vomit and feces, abdominal pain, inguinal pain, lack of appetite, hair loss, excessive cleaning behavior, extreme irritability, and distractibility. The larvae observed typically had the morphology of the larvae of insects related to the Cantharidae family in the Coleoptera order. For treatment, a single dose of albendazole (400 mg) was used. Consequently, in the present case, it was seen that the larvae of Coleoptera incidentally taken orally could continue to live for a period in the digestive tract of people, without losing vitality, and the larvae caused a variety of symptoms due to both their toxic agents and the possible irritation they caused.


Intraocular lens implants for uniocular cataracts in childhood.
We report the outcome of intraocular lens implantation in 20 children with visually significant cataracts (seven traumatic, 13 non-traumatic). Six patients had anterior and 14 had posterior chamber implants. The mean age of the whole group at the time of surgery was 5.9 years (range 0.3 to 15.1 years), while the mean period of follow-up was 2.4 years (range 0.8 to 5.9 years). Postoperatively 10 patients developed a transient fibrinous uveitis, four required lens repositioning, one needed lens removal, and eight required posterior capsulotomy. Complications warranting secondary surgical procedures occurred predominantly in eyes with posterior chamber implants. Co-operation with conventional amblyopia treatment was satisfactory in eight out of 16 patients. Postoperatively nine out of 18 patients had peripheral fusion, four patients regained visual acuities of better than 6/9, and visual acuity did not improve beyond 3/60 in six cases. In 19 eyes the optical pathway to the retina is clear and the implants are stable with no evidence of persisting inflammation. Contact lenses remain the initial treatment of choice in infancy, but modern intraocular lenses are well tolerated and have a role in the visual rehabilitation of patients with contact lens and probable contact lens failures and older children with uniocular cataracts.


Effect of oral rehydration solution versus spring water intake during exercise in the heat on muscle cramp susceptibility of young men.
Muscle cramp is a painful, involuntary muscle contraction, and that occurs during or following exercise is referred to as exercise-associated muscle cramp (EAMC). The causes of EAMC are likely to be multifactorial, but dehydration and electrolytes deficits are considered to be factors. This study tested the hypothesis that post-exercise muscle cramp susceptibility would be increased with spring water ingestion, but reduced with oral rehydration solution (ORS) ingestion during exercise. Ten men performed downhill running (DHR) in the heat (35-36 °C) for 40-60 min to reduce 1.5-2% of their body mass in two conditions (spring water vs ORS) in a cross-over design. The body mass was measured at 20 min and every 10 min thereafter during DHR, and 30 min post-DHR. The participants ingested either spring water or ORS for the body mass loss in each period. The two conditions were counter-balanced among the participants and separated by a week. Calf muscle cramp susceptibility was assessed by a threshold frequency (TF) of an electrical train stimulation to induce cramp before, immediately after, 30 and 65 min post-DHR. Blood samples were taken before, immediately after and 65 min after DHR to measure serum sodium, potassium, magnesium and chroride concentrations, hematocrit (Hct), hemoglobin (Hb), and serum osmolarity. Changes in these varaibles over time were compared between conditions by two-way repeated measures of analysis of variance. The average (±SD) baseline TF (25.6 ± 0.7 Hz) was the same between conditions. TF decreased 3.8 ± 2.7 to 4.5 ± 1.7 Hz from the baseline value immediately to 65 min post-DHR for the spring water condition, but increased 6.5 ± 4.9 to 13.6 ± 6.0 Hz in the same time period for the ORS condition (P < 0.05). Hct and Hb did not change significantly (P > 0.05) for both conditions, but osmolarity decreased (P < 0.05) only for the spring water condition. Serum sodium and chloride concentrations decreased (< 2%) at immediately post-DHR for the spring water condition only (P < 0.05). These results suggest that ORS intake during exercise decreased muscle cramp susceptibility. It was concluded that ingesting ORS appeared to be effective for preventing EAMC.


An oligomer study on small band gap polymers.
Small band gap polymers may increase the energy conversion efficiency of polymer solar cells by increased absorption of sunlight. Here we present a combined experimental and theoretical study on the optical and electrochemical properties of a series of well-defined, lengthy, small band gap oligo(5,7-bis(thiophen-2-yl)thieno[3,4-b]pyrazine)s ( E g = 1.50 eV) having alternating donor and acceptor units. The optical absorptions of the ground state, triplet excited state, radical cation, and dication are identified and found to shift to lower energy with increasing chain length. The reduction of the band gap in these alternating small band gap oligomers mainly results from an increase of the highest occupied molecular orbital (HOMO) level. The S 1-T 1 singlet-triplet splitting is reduced from approximately 0.9 eV from the trimeric monomer to -0.5 eV for the pentamer. This significant exchange energy is consistent with the fact that both the HOMO and the lowest unoccupied molecular orbital (LUMO) remain distributed over virtually all units, rather than being localized on the D and A units.


Effect of age on the responsiveness of vascular alpha-adrenoceptors in man.
Aging has been reported to alter the responsiveness of beta-receptors, but there have been few comparable studies of alpha-receptors. This study compares in six young and six healthy elderly subjects the haemodynamic effects of the alpha 1 -antagonist prazosin and the pressor responses to the alpha 1-agonist phenylephrine. Prazosin orally lowered erect (but not supine) blood pressure in both groups by a similar amount between 2 and 6 h after dosing. Maximal falls in systolic pressure were 19.5 +/- 15.7 and 29.3 +/- 11.4 mm Hg (mean +/- SD) in young and old, respectively. There was a significant difference in the associated heart rate response: in the young group mean heart rate increased to 103 beats/min, but there was no corresponding increase in the elderly group, which has a mean heart rate of 80 beats/min. Following the intravenous infusion of increasing doses of phenylephrine, log dose-response curves were deprived, and the dose required to raise mean arterial pressure by 20 mm Hg (PD20) was compared. The mean PD20 was significantly different in the two groups: 2.5 +/- 1.6 in the young, compared to 4.6 +/- 2.3 micrograms/kg/min in the elderly. However, no significant difference was apparent when pressor responsiveness was determined following the administration of prazosin. Thus, while there is no evidence of an age-related increase in the sensitivity of alpha-adrenoceptor-mediated vasoconstriction, the data are not inconsistent with an age-related reduction in responsiveness to alpha-adrenoceptor activation.


Tinnitus: animal models and findings in humans.
Chronic tinnitus (ringing of the ears) is a medically untreatable condition that reduces quality of life for millions of individuals worldwide. Most cases are associated with hearing loss that may be detected by the audiogram or by more sensitive measures. Converging evidence from animal models and studies of human tinnitus sufferers indicates that, while cochlear damage is a trigger, most cases of tinnitus are not generated by irritative processes persisting in the cochlea but by changes that take place in central auditory pathways when auditory neurons lose their input from the ear. Forms of neural plasticity underlie these neural changes, which include increased spontaneous activity and neural gain in deafferented central auditory structures, increased synchronous activity in these structures, alterations in the tonotopic organization of auditory cortex, and changes in network behavior in nonauditory brain regions detected by functional imaging of individuals with tinnitus and corroborated by animal investigations. Research on the molecular mechanisms that underlie neural changes in tinnitus is in its infancy and represents a frontier for investigation.


The proportion of dyslipidemia in systemic lupus erythematosus patient and distribution of correlated factors.
To understand the proportion of dyslipidemia in systemic lupus erythematosus (SLE) patients and the influencing factors of dyslipidemia. AN observational, cross-sectional study was conducted on new and longstanding SLE patients who had been diagnosed based on ARA criteria 1982 with 1997 revision. They had been hospitalized and treated at Department of Internal Medicine, Cipto Mangunkusumo National Central General Hospital and the other private Hospitals in Jakarta, i.e. Kramat Hospital in July - November 2003. The sample was selected by non probability sampling method with consecutive sampling technique. Every participant underwent history taking, physical and laboratory examination. There were 77 patients satisfying the inclusion criteria. The proportion of dyslipidemia in this study was 75.3%. By confidence interval of 95%, the dyslipidemia in SLE patient was 65.3% - 84.6%. The distribution of lipid profile in sample population were 43% with total cholesterol > or = 200 mg/dL, 26% with HDL cholesterol level < 40 mg/dL, 26.4% with LDL cholesterol level > or = 130 mg/dl and 44.2% with triglycerides serum level > or = 150 mg/dL. The characteristics of influencing factors in dyslipidemia prevalence for sample population consisted of 24.7% with renal involvement, 53.2% with > or = 3 years illness periods, 26% had received > or = 30 mg/day prednisone, 94.8% had not received chloroquines, and 58.4% had illness activity of Mex-SLEDAI > or = 2. By bivariate analysis, we found that illness period < 3 years tends to affect dyslipidemia with OR value of 12.04 (CI 95%, 2.54-57.05, p = 0.001). After conducting multivariate analysis by backward methods, it appears that only one significant influencing factor of dyslipidemia prevalence in SLE patient i.e. Illness period od < 3 years with OR value 12.04 (CI 95% 2.54 - 57.05, p = 0.001). Illness period of 3 years is represent a significant correlative factor for dyslipedemia prevalence. Prednisone > or = 30 mg/dL is the correlative factor for total cholesterol > or = 200 mg.dL and triglycerides > or = 150 mg/dL. Mex-SLEDAI > or = 2 is the corrective factor for HDL cholesterol < 40 mg/dL.


Cytosolic pH regulation in osteoblasts. Interaction of Na+ and H+ with the extracellular and intracellular faces of the Na+/H+ exchanger.
The interaction of Na and H ions with the extracellular and intracellular sites of the Na+/H+ exchanger of the osteosarcoma cell line UMR-106 was investigated. Na ions interact with a single, saturable extracellular transport site. H+ and amiloride appear to compete with Na+ for binding to this site. The apparent affinity for extracellular Na+ (Nao+) and amiloride was independent of intracellular H+ (Hi+), Nai+, or an outwardly directed H+ gradient. The interaction of H+ with the intracellular face of the exchanger had a sigmoidal characteristic with a Hill coefficient of approximately 2. The apparent affinity for Hi+ was independent of Nao+ between 25 and 140 mM. The apparent affinity for Hi+, but not the number of intracellular sites, increased with the increase in the outwardly directed H+ gradient across the membrane. Nai+/Ho+ exchange (reverse mode) is an electroneutral process with a Na+/H+ stoichiometry of 1. The dependence of Nai+/Ho+ exchange on Nai+ was sigmoidal, with a Hill coefficient of 2.16. Nai+ competes with Hi+ for binding to at least the transport site. The apparent affinity for Nai+ decreased with the increase in the outwardly directed H+ gradient. High Ho+ inhibited exchange activity in the reverse mode. We conclude that intracellular Na+ and H+ can activate the exchanger. The exchanger has two separate and asymmetric extracellular and intracellular transport sites. The relative apparent affinities of the internal transport site for Na+ and H+ are determined by the direction and magnitude of the H+ gradient across the membrane. Kinetic characterization of the exchanger suggests that Na+/H+ exchange is compatible with a simultaneous transport model, although a ping-pong transport model could not be excluded.


Metapopulation dynamics of the mistletoe and its host in savanna areas with different fire occurrence.
Mistletoes are aerial hemiparasitic plants which occupy patches of favorable habitat (host trees) surrounded by unfavorable habitat and may be possibly modeled as a metapopulation. A metapopulation is defined as a subdivided population that persists due to the balance between colonization and extinction in discrete habitat patches. Our aim was to evaluate the dynamics of the mistletoe Psittacanthus robustus and its host Vochysia thyrsoidea in three Brazilian savanna areas using a metapopulation approach. We also evaluated how the differences in terms of fire occurrence affected the dynamic of those populations (two areas burned during the study and one was fire protected). We monitored the populations at six-month intervals. P. robustus population structure and dynamics met the expected criteria for a metapopulation: i) the suitable habitats for the mistletoe occur in discrete patches; (ii) local populations went extinct during the study and (iii) colonization of previously non-occupied patches occurred. The ratio of occupied patches decreased in all areas with time. Local mistletoe populations went extinct due to two different causes: patch extinction in area with no fire and fire killing in the burned areas. In a burned area, the largest decrease of occupied patch ratios occurred due to a fire event that killed the parasites without, however, killing the host trees. The greatest mortality of V. thyrsoidea occurred in the area without fire. In this area, all the dead trees supported mistletoe individuals and no mortality was observed for parasite-free trees. Because P. robustus is a fire sensitive species and V. thyrsoidea is fire tolerant, P. robustus seems to increase host mortality, but its effect is lessened by periodic burning that reduces the parasite loads.


CD5L is a canonical component of circulatory IgM.
Immunoglobulin M (IgM) is an evolutionary conserved key component of humoral immunity, and the first antibody isotype to emerge during an immune response. IgM is a large (1 MDa), multimeric protein, for which both hexameric and pentameric structures have been described, the latter additionally containing a joining (J) chain. Using a combination of single-particle mass spectrometry and mass photometry, proteomics, and immunochemical assays, we here demonstrate that circulatory (serum) IgM exclusively exists as a complex of J-chain-containing pentamers covalently bound to the small (36 kDa) protein CD5 antigen-like (CD5L, also called apoptosis inhibitor of macrophage). In sharp contrast, secretory IgM in saliva and milk is principally devoid of CD5L. Unlike IgM itself, CD5L is not produced by B cells, implying that it associates with IgM in the extracellular space. We demonstrate that CD5L integration has functional implications, i.e., it diminishes IgM binding to two of its receptors, the FcαµR and the polymeric Immunoglobulin receptor. On the other hand, binding to FcµR as well as complement activation via C1q seem unaffected by CD5L integration. Taken together, we redefine the composition of circulatory IgM as a J-chain containing pentamer, always in complex with CD5L.


Pore formation: an ancient yet complex form of attack.
Bacteria, as well as higher organisms such as sea anemones or earthworms, have developed sophisticated virulence factors such as the pore-forming toxins (PFTs) to mount their attack against the host. One of the most fascinating aspects of PFTs is that they can adopt a water-soluble form at the beginning of their lifetime and become an integral transmembrane protein in the membrane of the target cells. There is a growing understanding of the sequence of events and the various conformational changes undergone by these toxins in order to bind to the host cell surface, to penetrate the cell membranes and to achieve pore formation. These points will be addressed in this review.


Early activation of matrix metalloproteinases underlies the exacerbated systolic and diastolic dysfunction in mice lacking TIMP3 following myocardial infarction.
Extracellular matrix (ECM) remodeling is a critical aspect of cardiac remodeling following myocardial infarction. Tissue inhibitors of metalloproteinases (TIMPs) are physiological inhibitors of matrix metalloproteinases (MMPs) that degrade the ECM proteins. TIMP3 is highly expressed in the heart, and is markedly downregulated in patients with ischemic cardiomyopathy. We therefore examined the time- and region-dependent role of TIMP3 in the cardiac response to myocardial infarction (MI). TIMP3(-/-) and wild-type (WT) mice were subjected to MI by ligation of the left anterior descending artery. TIMP3(-/-)-MI mice exhibited a significantly compromised rate of survival compared with WT-MI mice, primarily due to increased left ventricular (LV) rupture, greater infarct expansion, exacerbated LV dilation, and greater systolic and diastolic dysfunction. Second harmonic generation imaging of unfixed and unstained hearts revealed greater collagen disarray and reduced density in the TIMP3(-/-) infarct myocardium compared with the WT group. Gelatinolytic and collagenolytic activities increased in TIMP3(-/-) compared with WT hearts at 1 day post-MI but not at 3 days or 1 wk post-MI. Neutrophil infiltration and inflammatory MMPs were significantly increased in the infarct and peri-infarct regions of TIMP3(-/-)-MI hearts. Treatment of TIMP3(-/-) mice with a broad-spectrum MMP inhibitor (PD-166793) for 2 days before and 2 days after MI markedly improved post-MI infarct expansion, LV rupture incident, LV dilation, and systolic dysfunction in these mice up to 1 wk post-MI. Our data demonstrate that the initial rise in proteolytic activities early post-MI is a triggering factor for subsequent LV adverse remodeling, LV rupture, and dilated cardiomyopathy. Hence, timing of treatments to improve cardiac response to MI may be critical in producing favorable outcome.


Protective effect of n-acetylcysteine in a model of influenza infection in mice.
Reactive oxygen intermediates (ROI) and cytokines, particularly tumor necrosis factor (TNF) have been implicated in the pathogenesis of influenza. Using a murine model of influenza, we have studied the levels of TNF, interleukin 6 (IL-6) and of superoxide-generating xanthine oxidase (XO). Mice infected intranasally with influenza virus APR/8 had high levels of XO, TNF and IL-6 in the broncoalveolar lavage, as early as 3 d after infection. XO was elevated also in serum and lung tissue. Administration of the antioxidant N-acetylcysteine (NAC,1 g/kg per day, orally) significantly decreased the mortality in infected mice, indicating a role for RO1 in the lethality associated with influenza infection.


Impairments of Visuospatial Attention in Children with Unilateral Spastic Cerebral Palsy.
This observational study aimed at assessing the prevalence of visuospatial attention deficits in children with unilateral spastic cerebral palsy (USCP), taking into consideration the affected hemibody and the localization of the brain lesion. Seventy-five children with USCP were assessed with four visuospatial attention tests: star cancellation, Ogden figure copy, line bisection, and proprioceptive pointing. A majority (64%) of children with USCP presented a deficit in at least one test compared to the reference values. The alterations observed in children with left or right USCP were related to egocentric or allocentric neglect, respectively. Children with cortico/subcortical lesion presented more often visuospatial attention deficits than children with periventricular lesion. Visuospatial attention deficits were not associated with brain lesion locations. Visuospatial attention deficits are prevalent in children with USCP and should be taken into account during their rehabilitation process. The present results shed new light on the interpretation of motor impairments in children with USCP as they may be influenced by the frequent presence of visuospatial deficits.


[The relationship among theory, research and practice in nursing].
The present study is an attempt to present a process of relationship among theory, research, and practice in nursing. First, the term theory is defined as well as each of the theory's basic elements. Secondly, the concepts of research and practice are discussed in terms of their potential for stemming, testing, and redefining the theory. Thirdly, the concepts of theory, research and practice are presented as a triple spiral which works as a dinamic process. Within this process the theory, research, and practice interact to improve each other.


Jejunal diverticular disease complicated by enteroliths: Report of two different presentations.
Jejunal diverticula are quite rare. Furthermore, small bowel diverticular disease resulting in enteroliths can lead to complications necessitating surgical intervention. In this manuscript, we report two presentations of jejunal diverticulum with complications from enteroliths followed by a review of the literature. The first case was that of a 79-year-old male who presented with abdominal pain and was found, on computed tomography (CT) scan, to have evidence of intestinal perforation. A laparotomy showed that he had perforated jejunal diverticulitis. The second case was that of an 89-year-old female who presented with recurrent episodes of bowel obstruction. A laparotomy showed that she had an enterolith impacted in her jejunum in the presence of significant diverticular disease. Although a rare entity, familiarity with jejunal diverticular disease, its complications, and its management, should be part of every surgeon's base of knowledge when considering abdominal pathology.


Influence of age and cardiovascular factors on regional pulsed wave Doppler myocardial imaging indices.
To describe the influence of age and other cardiovascular factors on regional pulsed wave Doppler myocardial imaging (DMI), and to compare DMI with conventional transmitral echocardiography and the atrioventricular plane displacement (AVPD) method. Eighty-eight healthy subjects aged 20-81 years were examined by DMI, performed in the intraventricular septum just below the mitral annulus and in the corresponding lateral region, by transmitral pulsed wave Doppler echocardiography, and by AVPD. The DMI peak velocity during the left ventricular (LV) early filling phase (e), decreased with age from 12.3 +/- 2.3 cm/s in the youngest to 7.0 +/- 1.7 cm/s in the oldest tercentile (r=- 0.76, P<0.001). The DMI peak velocity during atrial contraction (a), increased from 7.5 +/- 2.2 cm/s in the youngest to 9.7 +/- 1.7 cm/s in the oldest tercentile (r=0.41, P<0.001). The DMI systolic peak velocity (s), decreased with age from 8.2 +/- 1.1 (youngest tercentile) to 6.9 +/- 1.1 (oldest tercentile), r=-0.39, P<0.001 cm/s, while the fraction shortening of the LV increased from 33.7 +/- 4.1 to 38.2 +/- 5.9% (r=0.36, P<0.01). The DMI e/a correlated with the transmitral early/atrial (E/A) (r=0.83, P<0.001) and with the AVPD measurement of diastolic function AV-LA/AV-mean (r=0.82, P<0.001). The DMI e velocity correlated with the transmitral E velocity (r=0.38, P<0.001). In the multiple regression analysis of DMI e, age was the strongest factor and LV mass index correlated inversely and independently with e. No DMI variables were influenced by gender, while transmitral E correlated with gender. The LV dimension variables explained 35% (R2 adjusted) of the DMI e velocity changes; only 7% of the transmitral E changes were explained by those variables. Regional DMI indices are highly age-dependent. In comparision with conventional echocardiography, regional DMI might be more influenced by LV geometry and by myocardial structural changes. These findings suggest a complementary role for regional DMI to conventional echocardiography for the assessment of myocardial function.


Enabling functional independence in Parkinson's disease: update on occupational therapy intervention.
Motor impairment and functional limitations are known sequelae of Parkinson's disease. Occupational therapy (OT) is recognized as an important adjunct to pharmacological management. This critical review provides an update on the effectiveness of OT. Eight studies were included in the review, three on occupational therapy task-related training, two on functional training with external cues and three on OT as part of interdisciplinary treatment. Due to lack of level I studies, it is difficult to determine conclusively the effect of OT. However, there is evidence to suggest that treatment produces improvements in motor and quality of life for the duration of therapy. Implications for future studies and practice are discussed.


The evolution, ecology, and conservation of hummingbirds and their interactions with flowering plants.
The ecological co-dependency between plants and hummingbirds is a classic example of a mutualistic interaction: hummingbirds rely on floral nectar to fuel their rapid metabolisms, and more than 7000 plant species rely on hummingbirds for pollination. However, threats to hummingbirds are mounting, with 10% of 366 species considered globally threatened and 60% in decline. Despite the important ecological implications of these population declines, no recent review has examined plant-hummingbird interactions in the wider context of their evolution, ecology, and conservation. To provide this overview, we (i) assess the extent to which plants and hummingbirds have coevolved over millions of years, (ii) examine the mechanisms underlying plant-hummingbird interaction frequencies and hummingbird specialization, (iii) explore the factors driving the decline of hummingbird populations, and (iv) map out directions for future research and conservation. We find that, despite close associations between plants and hummingbirds, acquiring evidence for coevolution (versus one-sided adaptation) is difficult because data on fitness outcomes for both partners are required. Thus, linking plant-hummingbird interactions to plant reproduction is not only a major avenue for future coevolutionary work, but also for studies of interaction networks, which rarely incorporate pollinator effectiveness. Nevertheless, over the past decade, a growing body of literature on plant-hummingbird networks suggests that hummingbirds form relationships with plants primarily based on overlapping phenologies and trait-matching between bill length and flower length. On the other hand, species-level specialization appears to depend primarily on local community context, such as hummingbird abundance and nectar availability. Finally, although hummingbirds are commonly viewed as resilient opportunists that thrive in brushy habitats, we find that range size and forest dependency are key predictors of hummingbird extinction risk. A critical direction for future research is to examine how potential stressors - such as habitat loss and fragmentation, climate change, and introduction of non-native plants - may interact to affect hummingbirds and the plants they pollinate.


Regulation of intracellular ceramide content in B16 melanoma cells. Biological implications of ceramide glycosylation.
We previously reported that ceramide released from glycosphingolipids (GSLs) by endoglycoceramidase was directly metabolized to GSLs, and thus the content of GSLs was constantly maintained in B16 melanoma cells (Ito, M., and Komori, H. (1996) J. Biol. Chem. 271, 12655-12660). In this study, the metabolism of ceramide released from sphingomyelin (SM) by bacterial sphingomyelinase (SMase) was examined using B16 cells and their GSL-deficient mutant counterpart GM95 cells. Treatment of B16 melanoma cells with bacterial SMase effectively hydrolyzed SM on the plasma membrane. Under these conditions, NeuAcalpha2,3Galbeta1, 4Glcbeta1,1ceramide was significantly increased. Interestingly, UDP-glucose:ceramide glucosyltransferase-1 (GlcT-1) activity and GSL synthesis, but not SM synthesis or sphingosine generation, were found to be up-regulated by SMase treatment. The up-regulation of GSL synthesis seemed to occur at both the transcriptional and post-translational steps of GlcT-1 synthesis. Accumulation of ceramide by bacterial SMase was much higher in GM95 cells than in the parental cells. When the enzyme was removed from the culture medium, the intracellular ceramide level in B16 cells, but not that in the mutant cells, normalized. No rapid restoration of SM in either of the cell lines was observed after removal of the enzyme. SMase treatment strongly inhibited DNA synthesis in GM95 cells but not that in B16 cells. In the presence of D-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol, an inhibitor of GlcT-1, SMase treatment markedly increased the ceramide content and thus inhibited DNA synthesis in B16 cells. Our study provides the first evidence that GlcT-1 functions to regulate the level of intracellular ceramide by glycosylation of the ceramide when it is present in excess.


Asymptomatic embolisation for prediction of stroke in the Asymptomatic Carotid Emboli Study (ACES): a prospective observational study.
Whether surgery is beneficial for patients with asymptomatic carotid stenosis is controversial. Better methods of identifying patients who are likely to develop stroke would improve the risk-benefit ratio for carotid endarterectomy. We aimed to investigate whether detection of asymptomatic embolic signals by use of transcranial doppler (TCD) could predict stroke risk in patients with asymptomatic carotid stenosis. The Asymptomatic Carotid Emboli Study (ACES) was a prospective observational study in patients with asymptomatic carotid stenosis of at least 70% from 26 centres worldwide. To detect the presence of embolic signals, patients had two 1 h TCD recordings from the ipsilateral middle cerebral artery at baseline and one 1 h recording at 6, 12, and 18 months. Patients were followed up for 2 years. The primary endpoint was ipsilateral stroke and transient ischaemic attack. All recordings were analysed centrally by investigators masked to patient identity. 482 patients were recruited, of whom 467 had evaluable recordings. Embolic signals were present in 77 of 467 patients at baseline. The hazard ratio for the risk of ipsilateral stroke and transient ischaemic attack from baseline to 2 years in patients with embolic signals compared with those without was 2.54 (95% CI 1.20-5.36; p=0.015). For ipsilateral stroke alone, the hazard ratio was 5.57 (1.61-19.32; p=0.007). The absolute annual risk of ipsilateral stroke or transient ischaemic attack between baseline and 2 years was 7.13% in patients with embolic signals and 3.04% in those without, and for ipsilateral stroke was 3.62% in patients with embolic signals and 0.70% in those without. The hazard ratio for the risk of ipsilateral stroke and transient ischaemic attack for patients who had embolic signals on the recording preceding the next 6-month follow-up compared with those who did not was 2.63 (95% CI 1.01-6.88; p=0.049), and for ipsilateral stroke alone the hazard ratio was 6.37 (1.59-25.57; p=0.009). Controlling for antiplatelet therapy, degree of stenosis, and other risk factors did not alter the results. Detection of asymptomatic embolisation on TCD can be used to identify patients with asymptomatic carotid stenosis who are at a higher risk of stroke and transient ischaemic attack, and also those with a low absolute stroke risk. Assessment of the presence of embolic signals on TCD might be useful in the selection of patients with asymptomatic carotid stenosis who are likely to benefit from endarterectomy. British Heart Foundation.


Metastatic adenocarcinoma of bladder presenting with skin and visceral metastasis.
A 62-year-old male patient presented with multiple subcutaneous swellings over neck, axilla and scapular region for last 3 months. Further evaluation revealed the presence of bladder mass, deranged kidney function tests with multiple rounded opacities in lung. The patient was stabilised with haemodialysis and bilateral percutaneous nephrostomy tube placement. The patient was planned for multiagent chemotherapy after stabilisation but died within 2 weeks due to progressive disease. Fine needle aspiration cytology (FNAC) from the above-mentioned skin and lymph nodes swellings was suggestive of adenocarcinoma. We herein report this rare case of a metastatic adenocarcinoma of the bladder with extensive skin and visceral metastasis. The presence of skin metastasis confers a poor prognostic sign to the patient. They are usually resistant to available treatment modalities. To our knowledge, this is the first reported case of adenocarcinoma metastatic to skin and viscera.


Turning the Light Switch on Binding: Prefrontal Activity for Binding and Retrieval in Action Control.
According to action control theories, responding to a stimulus leads to the binding of response and stimulus features into a common representation, that is, an event file. Repeating any component of an event file retrieves all previously bound information, leading to performance costs for partial repetitions measured in so-called binding effects. Although otherwise robust and stable, binding effects are typically completely absent in "localization tasks," in which participants localize targets with spatially compatible responses. Yet, it is possible to observe binding effects in such when location features have to be translated into response features. We hypothesized that this modulation of binding effects is reflected in task involvement of the dorsolateral pFC (DLPFC). Participants localized targets with either direct (i.e., spatially compatible key) or translated (i.e., diagonally opposite to the spatially compatible key) responses. We measured DLPFC activity with functional near-infrared spectroscopy. On the behavioral level, we observed binding effects in the translated response condition, but not in the direct response condition. Importantly, prefrontal activity was also higher in the translated mapping condition. In addition, we found some evidence for the strength of the difference in binding effects in behavioral data being correlated with the corresponding effects in prefrontal activity. This suggests that activity in the DLPFC reflects the amount of executive control needed for translating location features into responses. More generally, binding effects seem to emerge only when the task at hand involves DLPFC recruitment.


Dependency Parser-based Negation Detection in Clinical Narratives.
Negation of clinical named entities is common in clinical documents and is a crucial factor to accurately compile patients' clinical conditions and to further support complex phenotype detection. In 2009, Mayo Clinic released the clinical Text Analysis and Knowledge Extraction System (cTAKES), which includes a negation annotator that identifies negation status of a named entity by searching for negation words within a fixed word distance. However, this negation strategy is not sophisticated enough to correctly identify complicated patterns of negation. This paper aims to investigate whether the dependency structure from the cTAKES dependency parser can improve the negation detection performance. Manually compiled negation rules, derived from dependency paths were tested. Dependency negation rules do not limit the negation scope to word distance; instead, they are based on syntactic context. We found that using a dependency-based negation proved a superior alternative to the current cTAKES negation annotator.


Myeloperoxidase Mediates Postischemic Arrhythmogenic Ventricular Remodeling.
Ventricular arrhythmias remain the leading cause of death in patients suffering myocardial ischemia. Myeloperoxidase, a heme enzyme released by polymorphonuclear neutrophils, accumulates within ischemic myocardium and has been linked to adverse left ventricular remodeling. To reveal the role of myeloperoxidase for the development of ventricular arrhythmias. In different murine models of myocardial ischemia, myeloperoxidase deficiency profoundly decreased vulnerability for ventricular tachycardia on programmed right ventricular and burst stimulation and spontaneously as assessed by ECG telemetry after isoproterenol injection. Experiments using CD11b/CD18 integrin-deficient (CD11b-/-) mice and intravenous myeloperoxidase infusion revealed that neutrophil infiltration is a prerequisite for myocardial myeloperoxidase accumulation. Ventricles from myeloperoxidase-deficient (Mpo-/-) mice showed less pronounced slowing and decreased heterogeneity of electric conduction in the peri-infarct zone than wild-type mice. Expression of the redox-sensitive gap junctional protein Cx43 (Connexin 43) was reduced in the peri-infarct area of wild-type compared with Mpo-/- mice. In isolated wild-type cardiomyocytes, Cx43 protein content decreased on myeloperoxidase/H2O2 incubation. Mapping of induced pluripotent stem cell-derived cardiomyocyte networks and in vivo investigations linked Cx43 breakdown to myeloperoxidase-dependent activation of matrix metalloproteinase 7. Moreover, Mpo-/- mice showed decreased ventricular postischemic fibrosis reflecting reduced accumulation of myofibroblasts. Ex vivo, myeloperoxidase was demonstrated to induce fibroblast-to-myofibroblast transdifferentiation by activation of p38 mitogen-activated protein kinases resulting in upregulated collagen generation. In support of our experimental findings, baseline myeloperoxidase plasma levels were independently associated with a history of ventricular arrhythmias, sudden cardiac death, or implantable cardioverter-defibrillator implantation in a cohort of 2622 stable patients with an ejection fraction >35% undergoing elective diagnostic cardiac evaluation. Myeloperoxidase emerges as a crucial mediator of postischemic myocardial remodeling and may evolve as a novel pharmacological target for secondary disease prevention after myocardial ischemia.


On the mechanism by which hormones induce the release of Ca2+ from mitochondria in the liver cell.
1. The abilities of dinitrophenol, NaCl, Ruthenium Red and the Ca2+-selective ionophore A23187 to release 45 Ca2+ from isolated hepatocytes and liver mitochondria (incubated at 37 degrees C in the presence of 0.1 microM-free Ca2+, Mg2+, ATP and phosphate ions) were compared with the action of adrenaline on 45Ca2+ release from isolated hepatocytes. The effects of adrenaline were most closely described by those of the ionophore A23187. 2. In isolated hepatocytes, a release of 45Ca2+ and stimulation of O2 utilization similar to that induced by adrenaline was observed in the presence of 500 and 20 microM-arachidonic acid respectively. The effect of arachidonic acid on 45Ca2+ release was not specific for this unsaturated fatty acid. 3. Inhibitors of arachidonic acid metabolism, including indomethacin and eicosa-5,811,14-tetraynoic acid, did not block the effects of adrenaline on 45Ca2+ or glucose release from isolated hepatocytes. 4. The ability of adrenaline to stimulate 45Ca2+ release from isolated hepatocytes was rapidly reversed after the subsequent addition of phenoxybenzamine to the cell suspension, and was completely blocked by 0.5 mM-dibucaine. 5. The results are consistent with the action of a Ca2+-selective ionophore in the mechanism by which adrenaline induces the release of Ca2+ from mitochondria in the liver cell and indicate that it is unlikely that arachidonic acid or a metabolite of arachidonic acid is involved in this process.


An RNA thermoswitch regulates daytime growth in Arabidopsis.
Temperature is a major environmental cue affecting plant growth and development. Plants often experience higher temperatures in the context of a 24 h day-night cycle, with temperatures peaking in the middle of the day. Here, we find that the transcript encoding the bHLH transcription factor PIF7 undergoes a direct increase in translation in response to warmer temperature. Diurnal expression of PIF7 transcript gates this response, allowing PIF7 protein to quickly accumulate in response to warm daytime temperature. Enhanced PIF7 protein levels directly activate the thermomorphogenesis pathway by inducing the transcription of key genes such as the auxin biosynthetic gene YUCCA8, and are necessary for thermomorphogenesis to occur under warm cycling daytime temperatures. The temperature-dependent translational enhancement of PIF7 messenger RNA is mediated by the formation of an RNA hairpin within its 5' untranslated region, which adopts an alternative conformation at higher temperature, leading to increased protein synthesis. We identified similar hairpin sequences that control translation in additional transcripts including WRKY22 and the key heat shock regulator HSFA2, suggesting that this is a conserved mechanism enabling plants to respond and adapt rapidly to high temperatures.


Secretory mechanisms. Behaviour of adenine nucleotides during the platelet release reaction induced by adenosine diphosphate and adrenaline.
1. Platelets containing adenine nucleotides labelled with (3)H and (14)C in vitro were aggregated biphasically with ADP and adrenaline. Amounts of ATP and ADP as well as the radioactivity of ATP, ADP, AMP, IMP, hypoxanthine and adenine were determined in platelets and plasma at different stages of aggregation. 2. ATP and ADP were released during the second aggregation phase and had a low specific radioactivity compared with the ATP and ADP retained by the cells. The specific radioactivity of intracellular nucleotides increased during release. The parameters observed with ADP and adrenaline as release inducers were the same as for collagen and thrombin. 3. Release induced by all four inducers was accompanied by conversion of cellular [(3)H]ATP into extracellular [(3)H]-hypoxanthine. By variation of temperature, inducer concentration, time after blood withdrawal and use of acetylsalicylic acid, the aggregation pattern caused by adrenaline and ADP could be made mono- or bi-phasic. Release or second-phase aggregation was intimately connected with the ATP-hypoxanthine conversion, whereas first phase aggregation was not. 4. The [(3)H]ATP-hypoxanthine conversion started immediately after ADP addition. With adrenaline it usually started with the appearance of the second aggregation phase. The conversion was present during first phase of ADP-induced aggregation only if a second phase were to follow. 5. When secondary aggregation took place while radioactive adenine was being taken up by the platelets, increased formation of labelled hypoxanthine still occurred, but there was either no change or an increase in the concentration of labelled ATP. 6. Biphasically aggregated platelets converted [(3)H]adenine more rapidly into [(3)H]-ATP and -hypoxanthine than non-aggregated platelets. Addition of [(3)H]adenine at different stages of biphasic aggregation showed that more [(3)H]hypoxanthine was formed during than after the release step. 7. We conclude that ADP and adrenaline, like thrombin and collagen, cause extrusion of non-metabolic granula-located platelet adenine nucleotides. During release metabolic ATP breaks down to hypoxanthine, and this process might reflect an ATP-requiring part of the release reaction.


Tsunami evacuation simulation using geographic information systems for homecare recipients depending on electric devices.
Tsunamis cause direct damage to property and destroy infrastructure. In addition, power outages can lead to death, especially for patients who rely on medical equipment requiring a power supply. Recently, Nankai Trough Earthquakes have been predicted, and much effort has been put into developing countermeasures in Japan. Kochi City on Shikoku Island is expected to suffer in the event of a large tsunami. The present study identifies individuals living in Kochi who need evacuation assistance and depend on electrical medical devices, simulates evacuation behavior and inundation during a tsunami using a geographic information system (GIS), and considers the usefulness of such a GIS. We asked caregivers, including visiting nurses, to introduce us to homecare recipients who rely on a ventilator, an endotracheal suction device, or other medical devices requiring electric power. We received introductions to 52 homecare recipients. Using a GIS, we plotted the area of predicted inundation and the locations of homecare recipients, nursing stations, and welfare evacuation shelters. We predicted evacuation routes, and then analyzed the time difference between the time required for evacuation and tsunami arrival at a welfare evacuation shelter. To measure the effects of the main parameters, we conducted both one-way and multi-way sensitivity analysis. In the event of a tsunami, eight of the homecare recipients living in the forecasted inundation areas in Kochi may face delayed evacuation. Among homecare recipients facing a high possibility of escape delay, 95.2% lived more than 1,800 m from the nearest welfare evacuation shelter. We found that individual evacuation behavior can be simulated by specifying the residence of a homecare recipient and the evacuation route using a GIS.


Retrograde enteroclysis by double balloon enteroscopy in a patient with blunt abdominal trauma: small bowel stricture, intraluminal vascular lesion and Crohn's disease.
A 40-year-old male presented to the Emergency Department after a driving accident with blunt abdominal trauma. An abdominal computed tomography (CT) scan revealed a mesenteric injury in the right lower quadrant. He was admitted two months later due to a one-day history of abdominal pain and diarrhea, without fever or blood. The CT angiography showed a pseudoaneurysm located in the proximal ileum and several rigid small bowel (SB) loops with segmental wall thickening of mucosa.


Establishing the differential diagnosis of neck lesions.
A thorough physical examination of the head and neck should be included as part of the evaluation of every dental patient. Although they most often represent a self-limited inflammation, a benign neoplasm, or a congenital anomaly, neck lesions can also be malignant. Because the pathologic lesions that can arise in the neck are varied and diverse, the diagnostic thought process should be orderly and inclusive so that a reasonable differential diagnosis can be established and prompt, effective treatment can be rendered.


Visual memory needs categories.
Capacity limitations in the way humans store and process information in working memory have been extensively studied, and several memory systems have been distinguished. In line with previous capacity estimates for verbal memory and memory for spatial information, recent studies suggest that it is possible to retain up to four objects in visual working memory. The objects used have typically been categorically different colors and shapes. Because knowledge about categories is stored in long-term memory, these estimations of working memory capacity have been contaminated by long-term memory support. We show that when using clearly distinguishable intracategorical items, visual working memory has a maximum capacity of only one object. Because attention is closely involved in the working memory process, our results add to other studies demonstrating capacity limitations of human attention such as inattentional blindness and change blindness.


Insertions, substitutions, and the origin of microsatellites.
This paper uses data from the Human Gene Mutation Database to contrast two hypotheses for the origin of short DNA repeats: substitutions and insertions that duplicate adjacent sequences. Because substitutions are much more common than insertions, they are the dominant source of new 2-repeat loci. Insertions are rarer, but over 70% of the 2-4 base insertion mutations are duplications of adjacent sequences, and over half of these generate new repeat regions. Insertions contribute fewer new repeat loci than substitutions, but their relative importance increases rapidly with repeat number so that all new 4-5-repeat mutations come from insertions, as do all 3-repeat mutations of tetranucleotide repeats. This suggests that the process of repeat duplication that dominates microsatellite evolution at high repeat numbers is also important very early in microsatellite evolution. This result sheds light on the puzzle of the origin of short tandem repeats. It also suggests that most short insertion mutations derive from a slippage-like process during replication.


Effect of infection with Trichobilharzia ocellata and Schistosoma mansoni on the ultrastructure of the albumen gland of their respective hosts, Lymnaea stagnalis and Biomphalaria glabrata.
The albumen gland, a female accessory sex gland of pulmonate snails, produces the perivitelline fluid. The ultrastructure of the albumen glands of control and infected specimens of Lymnaea stagnalis and Biomphalaria glabrata was studied. The albumen gland of L. stagnalis contains two types of secretory cells--light (active) and dark (inactive)--and two types of supporting cells--centroacinar and myoepithelial. The secretory cells apparently represent two activity stages of one type of cell. The gland B. glabrata possesses only one secretory cell type, which alternates with one type of supporting cell. The albumen glands of L. stagnalis and B. glabrata infected at a juvenile stage were studied 4 and 14 weeks (L. stagnalis) and 4 and 9 weeks (B. glabrata) after exposure. After four weeks' infection, B. glabrata produced some egg masses, but in subsequent stages egg mass production completely coased. Infected L. stagnalis never produced eggs. B. glabrata was apparently infected at a "physiologically" more mature stage than L. stagnalis. The morphology of the albumen glands four weeks after exposure (the daughter sporocyst stage) is in agreement with this hypothesis. At this interval the secretory cells of L. stagnalis appeared to be much more severely affected (inactive Golgi bodies and rough endoplasmic reticulum, crinophagy of the secretory granules) than the cells of B. glabrata. In the later stages studied (shedding of the cercariae), the glands of both species appeared to be completely inactive (reduced height of the epithelium, inactive organelles, crinophagy, absence of secretory granules). At this stage of infection, daughter sporocysts containing cercaria embryos were seen in the connective tissue of the albumen gland of B. glabrata, but not of L. stagnalis. The results thus indicate that the development and synthetic activity of the albumen gland are seriously affected by infection. These processes are known to be under the endocrine control of the female gonadotrophic hormones. Since it has been established that these hormones are normally present in the haemolymph of infected snails, the findings can be explained by assuming that the parasite interferes in some way or other with the snail's endocrine system.


Direct measurement of helical cell motion of the spirochete leptospira.
Leptospira are spirochete bacteria distinguished by a short-pitch coiled body and intracellular flagella. Leptospira cells swim in liquid with an asymmetric morphology of the cell body; the anterior end has a long-pitch spiral shape (S-end) and the posterior end is hook-shaped (H-end). Although the S-end and the coiled cell body called the protoplasmic cylinder are thought to be responsible for propulsion together, most observations on the motion mechanism have remained qualitative. In this study, we analyzed the swimming speed and rotation rate of the S-end, protoplasmic cylinder, and H-end of individual Leptospira cells by one-sided dark-field microscopy. At various viscosities of media containing different concentrations of Ficoll, the rotation rate of the S-end and protoplasmic cylinder showed a clear correlation with the swimming speed, suggesting that these two helical parts play a central role in the motion of Leptospira. In contrast, the H-end rotation rate was unstable and showed much less correlation with the swimming speed. Forces produced by the rotation of the S-end and protoplasmic cylinder showed that these two helical parts contribute to propulsion at nearly equal magnitude. Torque generated by each part, also obtained from experimental motion parameters, indicated that the flagellar motor can generate torque >4000 pN nm, twice as large as that of Escherichia coli. Furthermore, the S-end torque was found to show a markedly larger fluctuation than the protoplasmic cylinder torque, suggesting that the unstable H-end rotation might be mechanically related to changes in the S-end rotation rate for torque balance of the entire cell. Variations in torque at the anterior and posterior ends of the Leptospira cell body could be transmitted from one end to the other through the cell body to coordinate the morphological transformations of the two ends for a rapid change in the swimming direction.


[Sunbathing habits and risk of melanoma].
The increasing frequency of malignant melanoma (MM) is believed mainly to be due to the habit of sunbathing, which has an impact on the skin due to ultraviolet radiation, especially if intermittent and causing sunburn. Frequent use of tanning booths possibly increases the risk of MM but the question has not been settled. Use of sunscreens has a certain prophylactic effect on actinic keratoses, but no documented effect on MM. This does not imply that sunscreens are not advisable, but stresses the importance of correct use of sunscreens together with other sun protective measures. Primary prevention of MM is mainly a question of minimizing the impact of ultraviolet radiation (including tanning booths) on the skin: limit time in the sun, cover up with clothing, and use a sunscreen with a protection factor of at least 15.


Preparation and Characterization of CuO/ZnO/PVDF/PAN Nanofiber Composites by Bubble-electrospinning.
Nanocomposites loaded with metal oxides, such as CuO and ZnO, have excellent optical, electrical, mechanical and chemical properties, which result in their great potential applications in optoelectronic devices, sensors, photocatalysts and other fields. Especially, electrospun metal- oxide-loaded nanofibers have attracted much attention in many fields. However, the single-needle Electrospinning (ES) inhibits the industrial application of these electrospun nanofiber composites. Bubble-Electrospinning (BE) is an effective free surface ES for mass production of nanofiber membranes loaded with metal oxide. Few relevant patents to the topic have been reviewed and introduced. The BE was used to prepare mass production of Cu(Ac)2 /Zn(Ac)2/ PVDF/ PAN Composite Nanofiber Membranes (CNFMs). Then PVDF/PAN CNFMs containing CuO and ZnO nanocrystals were obtained by heat-treatment. Finally, CuO nanosheets and ZnO nanorods were successfully grown on the surface of PVDF/PAN CNFMs using hydrothermal method. In addition, the morphology and crystal structure of CNFMs were investigated by scanning electron microscopy (SEM) and X-Ray Powder Diffractometer (XRD). The morphology and crystal structure of the samples were characterized by SEM and XRD. The results showed the heat treatment temperature of 150oC and the hydrothermal temperature of 150oC were the optimal process parameters for the fabrication of PVDF/PAN CNFMs loaded with CuO and ZnO nanocrystals, and a higher heat treatment temperature results in higher crystallinity of ZnO and CuO. CuO/ZnO/PVDF/PAN CNFMs were successfully prepared by a combination of BE, heattreatment and hydrothermal method. The ZnO/CuO beads obtained by heat treatment is the key point of growing ZnO/CuO nanocrystals, and the growth temperature has great effect on the morphology of ZnO/CuO nanocrystals.


Clinical and laboratory features of viral hepatitis A in children.
Recent outbreaks of viral hepatitis A in non-endemic European countries and the potential outbreak risk in susceptible populations has led us to evaluate the clinical characteristics of children hospitalised with hepatitis A. Retrospective study included 118 children (68 boys and 50 girls) with the mean age of 8.5 years hospitalised at Hospital Na Bulovce in Prague from June 2008 to June 2009. The clinical course was symptomatic icteric in 57 (48.3 %) children, symptomatic anicteric in 23 (19.5 %), subclinical in 22 (18.6 %) and asymptomatic inapparent in 16 (13.6 %). The relapse of the disease occurred in three patients. There were no cases of fulminant hepatitis. The most frequent symptoms included jaundice (57 cases), abdominal pain/discomfort (38), vomiting (38), dark urine (37), subfebrility (29) and fever (25). Hepatic injury markers were substantially elevated in icteric patients, but moderate elevations were identified in anicteric and subclinical cases as well. Lower white blood cell and lymphocyte counts were independently associated with symptomatic and more severe clinical course. Active immunisation was provided to 22 patients, and as a post-exposure prophylaxis to 19 of them. The clinical course and laboratory parameters in vaccinated children were not significantly different from non-vaccinated children. The clinical course of hepatitis A was largely self-limiting and benign. Asymptomatic infections are frequent, representing risk for disease spread; however, they are associated with elevations of hepatic injury markers. The inclusion of significant proportion of asymptomatic cases that were identified and investigated only because of active epidemiological surveillance in the outbreak focus represents the particular asset of this study.


Antibody-mediated rejection of human orthotopic liver allografts. A study of liver transplantation across ABO blood group barriers.
A clinicopathologic analysis of liver transplantation across major ABO blood group barriers was carried out 1) to determine if antibody-mediated (humoral) rejection was a cause of graft failure and if humoral rejection can be identified, 2) to propose criteria for establishing the diagnosis, and 3) to describe the clinical and pathological features of humoral rejection. A total of 51 (24 primary) ABO-incompatible (ABO-I) liver grafts were transplanted into 49 recipients. There was a 46% graft failure rate during the first 30 days for primary ABO-I grafts compared with an 11% graft failure rate for primary ABO compatible (ABO-C), crossmatch negative, age, sex and priority-matched control patients (P less than 0.02). A similarly high early graft failure rate (60%) was seen for nonprimary ABO-I grafts during the first 30 days. Clinically, the patients experienced a relentless rise in serum transaminases, hepatic failure, and coagulopathy during the first weeks after transplant. Pathologic examination of ABO-I grafts that failed early demonstrated widespread areas of geographic hemorrhagic necrosis with diffuse intraorgan coagulation. Prominent arterial deposition of antibody and complement components was demonstrated by immunoflourescent staining. Elution studies confirmed the presence of tissue-bound, donor-specific isoagglutinins within the grafts. No such deposition was seen in control cases. These studies confirm that antibody mediated rejection of the liver occurs and allows for the development of criteria for establishing the diagnosis.


Discriminative stimulus properties of cocaine: enhancement by monoamine reuptake blockers.
In this study we examined the ability of compounds varying in their in vitro potencies as inhibitors of dopamine (DA), norepinephrine (NE) or serotonin (5-HT) reuptake to enhance the discriminative stimulus (DS) effects of cocaine. Compounds were administered in combination with cocaine (2.5 mg/kg i.p.) to rats trained to discriminate a low dose from a high dose of cocaine (2.5 vs. 10 mg/kg i.p.) in a two-lever, FR10 drug discrimination paradigm. All the monoamine reuptake blockers produced high-dose-appropriate responding in a dose-related manner when combined with a low dose of cocaine, but compounds from other pharmacological classes (benztropine, caffeine, diazepam, or 8-hydroxy-2-(di-n-propylamino)tetralin) did not enhance the DS effects of cocaine. Analysis of the relationship between behavioral and in vitro biochemical potencies indicated that inhibition of DA and 5-HT transport is responsible for the cocaine-enhancing effects of the monoamine reuptake blockers we examined. In contrast, NE reuptake apparently does not play a strong role, despite the finding that desipramine, talsupram and nortriptyline enhanced the DS effects of cocaine. However, pretreatment with the alpha-1 adrenergic antagonist prazosin failed to alter completely the ability of desipramine to enhance the DS effects of the low training dose of cocaine, but did produce dose-related decreases in the cocaine-enhancing effects of the beta adrenergic antagonist propranolol (10 mg/kg i.p.). These findings suggested that, under some conditions, NE interactions can modulate the DS effects of cocaine. In all, the results confirm reports that monoamine reuptake blockers enhance the DS effects of cocaine and indicate that 5-HT and DA can effectively modulate the DS effects of cocaine, but suggest that NE interactions may be relatively less important in the rat.


Alginate-encapsulated producer cells: a potential new approach for the treatment of malignant brain tumors.
In recent years gene therapy has evolved as a new treatment for brain tumors, where genetically engineered cells can be used to deliver specific substances to target cells. However, clinical success has been limited due to insufficient gene transfer, lack of prolonged gene expression, and immunorejection of producer cells. These obstacles may be overcome by encapsulating producer cells into immunoisolating substances such as alginate. This may provide a stable in situ delivery system of specific proteins, which can interfere with tumor growth and differentiation. This article represents a fundamental study describing the in vitro and the in vivo behavior of alginate-encapsulated producer cells. The viability and cell cycle distribution of encapsulated NIH 3T3 cells was studied by confocal laser scanning microscopy (CLSM) and by flow cytometry. The CLSM study showed a high viability of the encapsulated NIH 3T3 cells during 9 weeks in culture. The flow cytometric analysis revealed a change in cellular ploidy after 1 week in culture, with normalization in ploidy after 3 and 9 weeks. The production of the bacterial E. coli beta-galactosidase in alginate-encapsulated BT4CnVlacZ cells was studied by x-gal staining, and the cells expressed prolonged beta-galactosidase activity. H528 hybridoma cells producing monoclonal antibodies (mAbs) against the human epidermal growth factor receptor (EGFR) were encapsulated in alginate, and the mAb release was determined. The release of mAbs stabilized around 400 ng/ml/h after 12 days in vitro. To actually demonstrate that alginate-encapsulated H528 cells potentially inhibit a heterogeneous glioma cell population, cell migration from human GaMg glioma spheroids was studied during stimulation with EGF in the presence of encapsulated H528 cells. The migration in vitro was totally inhibited in the presence of H528 encapsulated cells. Alginate beads with H528 cells were also implanted into rat brains, and after 9 weeks the distribution of mAbs within the brain was studied by immunohistochemistry. It is shown that the alginate entrapped H528 cells produce mAbs inside the brain for prolonged periods and that the mAbs are distributed within all CSF compartments. Encapsulated producer cells represent a potential delivery system for specific proteins to brain tumors. Different producer cells may be encapsulated in alginate to target phenotypic features and microenvironmental factors, which may influence the progressive growth of brain tumors.


The implications of upper-airway obstruction on successful infant feeding.
Oral feeding in infants requires highly integrated sucking, swallowing, and respiratory sequencing controlled by the neurologic system. Rapid neuromuscular coordination of oral, pharyngeal, and esophageal phases of swallowing must be coordinated with respiration in the swallowing process. When obstruction is present in the upper airway secondary to anatomic or physiologic anomalies, disruption to the oral feeding process may occur. The infant will likely be unable to coordinate sucking and swallowing with breathing in an advantageous sequence. Inefficient feeding and difficulty with airway protection during swallowing may have serious implications regarding the infant's respiratory health as well as ability to gain weight adequately. A stable and patent airway is always the first priority in the management of the infant with upper-airway obstruction. Evaluation of the infant's potential for oral feeding may occur simultaneously with the initial evaluation of the degree of airway obstruction or it may occur following medical or surgical intervention for the airway obstruction. The evaluation process and management options for oral feeding will depend upon the method used to establish a patent airway. Clinical assessment by the speech-language pathologist includes a thorough assessment of oral sensory and motor mechanics, recognition of clinical signs and symptoms of swallowing dysfunction, and consideration of referral for instrumental assessment to obtain objective information regarding airway protection and swallowing function. A variety of medical and feeding interventions may be used to help support oral feeding to whatever extent is safe and efficient. Infants with significant oral feeding problems in the presence of airway obstruction may require a period of supplemental tube feeding and non-nutritive oral stimulation.


Effects of magnesium sulfate on neuron apoptosis and expression of caspase-3, bax and bcl-2 after cerebral ischemia-reperfusion injury.
To investigate the effects of magnesium sulfate on neuron apoptosis and the expressions of caspase-3, bax and bcl-2 after cerebral ischemia-reperfusion injury. Thirty-six gerbils were randomly divided into three groups: Sham-operation group (So, n = 12), ischemia-reperfusion group (I-R, n = 12) and magnesium sulfate group (Ms, n = 12). The neuron apoptosis was detected by the terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-fluorescence nick end labeling (TUNEL stain), and the protein expressions of caspase-3, bax and bcl-2 were detected by immunohistochemisty stain. Apoptotic neurons and the expressions of caspase-3 and bax were significantly increased in I-R and Ms groups, compared with those in So group. Apoptotic neurons and the expressions of bax and caspase-3 in Ms group were significantly less than those in I-R group. There was no significant difference in the expression of bcl-2 among three groups. Magnesium sulfate decreases neuron apoptosis after cerebral ischemia-reperfusion injury, which may be related with the suppressed expressions of caspase-3 and bax.


Opposing chemosensory functions of closely related gustatory receptors.
In the fruit fly Drosophila melanogaster, gustatory sensory neurons express taste receptors that are tuned to distinct groups of chemicals, thereby activating neural ensembles that elicit either feeding or avoidance behavior. Members of a family of ligand -gated receptor channels, the Gustatory receptors (Grs), play a central role in these behaviors. In general, closely related, evolutionarily conserved Gr proteins are co-expressed in the same type of taste neurons, tuned to chemically related compounds, and therefore triggering the same behavioral response. Here, we report that members of the Gr28 subfamily are expressed in largely non-overlapping sets of taste neurons in Drosophila larvae, detect chemicals of different valence, and trigger opposing feeding behaviors. We determined the intrinsic properties of Gr28 neurons by expressing the mammalian Vanilloid Receptor 1 (VR1), which is activated by capsaicin, a chemical to which wild-type Drosophila larvae do not respond. When VR1 is expressed in Gr28a neurons, larvae become attracted to capsaicin, consistent with reports showing that Gr28a itself encodes a receptor for nutritious RNA. In contrast, expression of VR1 in two pairs of Gr28b.c neurons triggers avoidance to capsaicin. Moreover, neuronal inactivation experiments show that the Gr28b.c neurons are necessary for avoidance of several bitter compounds. Lastly, behavioral experiments of Gr28 deficient larvae and live Ca2+ imaging studies of Gr28b.c neurons revealed that denatonium benzoate, a synthetic bitter compound that shares structural similarities with natural bitter chemicals, is a ligand for a receptor complex containing a Gr28b.c or Gr28b.a subunit. Thus, the Gr28 proteins, which have been evolutionarily conserved over 260 million years in insects, represent the first taste receptor subfamily in which specific members mediate behavior with opposite valence.


Conflict and natural disaster: the impacts on medical education in Ukraine and Türkiye.
In addition to causing enormous and enduring harms to the health of populations, the Russian invasion of Ukraine and the recent earthquakes in southeast Türkiye have greatly damaged the institutions of medical education at work in these countries. This paper explores these harms and encourages medical educationalists in unaffected countries to reflect on the virtues of their own educational institutions.


"Heely"-related injuries in children.
"Heelys", or shoes with an integral wheel embedded into the heel, are becoming increasingly popular among children in the UK. Despite the manufacturer's claims about their safety, increasing numbers of patients are attending the emergency department with "Heely"-related injuries. To assess the number and type of "Heely"-related injuries seen in the emergency department in a busy district general hospital and to assess the number of school days lost as a result of these injuries as a secondary measure of the impact on education and lifestyle. Medical staff working in the emergency department completed proformas for all children attending the department with "Heely"-related injuries between 26 December and 26 April 2007. Data collected included age, sex, mechanism of injury, diagnosis and number of days off school as a result of the injury. 35 patients with "Heely"-related injuries of mean age 9.6 years (range 6-15) were identified during the study period. The most common mechanism of injury was a fall onto an outstretched hand (20/35, 57%). Other mechanisms of injury identified were lateral upper limb injury (7/35), traumatic lower limb injury (2/35), rotational lower limb injury (2/35), head injury (2/35) and back injury (2/35). The most common diagnosis was fracture of the distal radius (10/35), two of which had an associated distal ulna fracture. Two tibial fractures and one nasal fracture were also seen. The average number of days off school was 4.5 days (range 0-20). None of the children included in this study were using safety equipment at the time of the injury. The number of "Heely"-related injuries seen in one department over a 4-month period suggests a much higher incidence of injuries than the 46/100,000 found by the manufacturers based on Consumer Product Safety Commission data in the USA. The discrepancy is almost certainly due to the reluctance of UK children to use safety equipment and to follow the manufacturer's safety advice. Larger scale studies are needed to confirm the incidence of "Heely"-related injuries in emergency departments across the UK. If the high incidence of injuries among users is confirmed, primary and secondary prevention methods could be considered to reduce injuries.


Toxicity of boric acid, borax and other boron containing compounds: A review.
Boron, often in the form of boric acid, is widely used as a flame retardant in insulation products, and although humans ingest boron through food, high exposure may lead to unwanted health effects. We assessed the toxicity of boric acid, borax and other forms of boron, after inhalation, dermal and oral exposure. After oral exposure, boron is absorbed over the gastrointestinal tract. Intact skin seems to pose a more effective barrier to boron than compromised skin. Boron excretion seems to mainly occur via the urine, although after skin exposure boron has been demonstrated in bile and gastrointestinal contents. Inhalation toxicity data are sparse, but one animal study showed reduced foetal weight after inhalation of cellulose that had a boric acid content of 20%. Skin exposure to boric acid has proven fatal in some cases, and the range of toxicity effects include abdominal as well as local effects on the skin. Fatalities from boric acid also have occurred after oral ingestion, and the endpoints in animals are weight loss and reproductive toxicity. Concerning genotoxicity studies, the overall picture indicates that boron-containing compounds are not genotoxic. There was no evidence of the carcinogenicity of boric acid in a 2-year study in mice.


Endothelial nitric oxide synthase-dependent cerebral blood flow augmentation by L-arginine after chronic statin treatment.
Nitric oxide, a product of nitric oxide synthase activity, relaxes vascular smooth muscle and elevates brain blood flow. We evaluated the importance of eNOS to cerebral blood flow augmentation after L-arginine infusion and increases in flow after eNOS upregulation in SV-129 mice. Blood flow was measured by laser-Doppler flowmetry before and after L-arginine infusion (450 mg/kg during a 15-minute period) or measured by 14C-iodoamphetamine indicator fractionation or 14C-iodoantipyrine tissue equilibration techniques. rCBF increased by 26% (laser Doppler flowmetry) after L-arginine infusion but did not change in mutant mice deficient in eNOS expression. After eNOS upregulation by chronic simvastatin treatment (2 mg/kg subcutaneously, daily for 14 days), L-arginine amplified and sustained the hyperemia (38%) and increased absolute brain blood flow from 86 +/- 7 to 119 +/- 10 mL/100 g per minute. Furthermore, pretreatment with simvastatin enhanced blood flow within ischemic brain tissue after middle cerebral artery occlusion. Together, these findings suggest that eNOS activity is critical for blood flow augmentation during acute L-arginine infusion, and chronic eNOS upregulation combined with L-arginine administration provides a novel strategy to elevate cerebral blood flow in the normal and ischemic brain.


Bioactive constituents of Conyza albida.
The bioactivity-guided fractionation of an active chloroform extract of Conyza albida led to the isolation of three alkenynes, deca-4,6-diyn-2-(Z)-enoic methyl ester (1), deca-4,6-diyn-2-(Z)-enoic ethyl ester (2) and deca-2,4-diene-4-hydroxy-6-yn-1,4-olide (3), and the terpenoid spathulenol (4), as the active toxic metabolites in the Artemia sp. lethality test. When tested in the KB cell cytotoxicity assay, compounds 1-4 demonstrated IC50 values of 52.2, 38.4, 117.9, and 83.8 microM, respectively. All compounds studied were inactive in the DNA methyl green and DNA strand scission assays, while compounds 3 and 4 showed moderate activity as inhibitors of human topoisomerase I. Compound 2 is reported here for the first time.


Towards comparable assessment of the soil nutrient status across scales-Review and development of nutrient metrics.
Nutrient availability influences virtually every aspect of an ecosystem, and is a critical modifier of ecosystem responses to global change. Although this crucial role of nutrient availability in regulating ecosystem structure and functioning has been widely acknowledged, nutrients are still often neglected in observational and experimental synthesis studies due to difficulties in comparing the nutrient status across sites. In the current study, we explain different nutrient-related concepts and discuss the potential of soil-, plant- and remote sensing-based metrics to compare the nutrient status across space. Based on our review and additional analyses on a dataset of European, managed temperate and boreal forests (ICP [International Co-operative Programme on Assessment and Monitoring of Air Pollution Effects on Forests] Forests dataset), we conclude that the use of plant- and remote sensing-based metrics that rely on tissue stoichiometry is limited due to their strong dependence on species identity. The potential use of other plant-based metrics such as Ellenberg indicator values and plant-functional traits is also discussed. We conclude from our analyses and review that soil-based metrics have the highest potential for successful intersite comparison of the nutrient status. As an example, we used and adjusted a soil-based metric, previously developed for conifer forests across Sweden, against the same ICP Forests data. We suggest that this adjusted and further adaptable metric, which included the organic carbon concentration in the upper 20 cm of the soil (including the organic fermentation-humus [FH] layer), the C:N ratio and pHCaCl2 of the FH layer, can be used as a complementary tool along with other indicators of nutrient availability, to compare the background nutrient status across temperate and boreal forests dominated by spruce, pine or beech. Future collection and provision of harmonized soil data from observational and experimental sites is crucial for further testing and adjusting the metric.