Exploring the electronic properties and cation complexation of polyoxoaurates [Au(III)4X4Om](n-) (X = Si(IV), P(V), Ge(IV), As(V), and Se(IV)) using quantum chemical calculations.
Quantum chemical calculations were performed to explore the structural and electronic properties of the two polyoxoaurates, [Au(III)4As(V)4O20](8-) (Au4As4) and [Au(III)4Se(IV)4O16](4-) (Au4Se4), known to date, and a number of hypothetical polyoxoaurate derivatives comprising heteroatoms different from arsenic and selenium (namely, Si, Ge and P). In addition, the interactions of [Au(III)4X4Om](n-) (X = As, Se) with alkali-metal cations (Li(+), Na(+), K(+) and Rb(+)) are also analysed. The studies suggest that the geometry structure, electronic properties and nucleophilicity of oxygen atoms of these polyoxoaurates are tuned by the size or electronegativity of the heteroatoms (Si, Ge, P, As and Se). Then, the geometry of [Au(III)4X4Om](n-) (X = As and Se) coordinating with alkali cations from Li(+) to Rb(+) and the complexation energy between [Au(III)4X4Om](n-) and alkali cations were compared. The results show that the stability and electronic structure of heteropolyoxoaurates depend on the entrapped cations. On the basis of the complexation energy, it can be concluded that the ion-pairing effect in arsenate-capped oxoaurate is stronger than that in selenite-capped oxoaurate. These heteropolyoxoaurates are expected to play a role in aqueous behaviour, self-assembly characteristics of polyoxoaurates, ion recognition, selectivity studies and may exhibit potential guest-switchable redox properties.


Serum zinc levels in subtypes of leprosy.
Serum zinc levels were estimated in 146 patients with different types of leprosy and 40 control subjects. Tuberculoid patients were found to have normal serum zinc levels. A gradual reduction in serum zinc levels was observed from TT to LL. Serum zinc levels were not significantly higher in patients treated with dapsone for approximately 18 months. The cause of hypozincemia is not clear, but the findings suggest that there are correlations among bacillary load, immune status, and serum zinc. Zinc deficiency could be one of the factors involved in the nonspecific suppression of cell-mediated immunity in lepromatous leprosy.


Patient outcome at long-term follow-up after aggressive microsurgical resection of cranial base chordomas.
In this study, we evaluated patients' clinical outcome and recurrence rates at long-term follow-up after aggressive microsurgical resection of cranial base chordomas. Seventy-four patients with chordomas underwent operations during a 16-year period from 1988 to 2004. The philosophy was to perform complete resection whenever possible and to provide adjuvant radiotherapy for remnants. Staged operations were performed for extensive tumors or if a sizable tumor remnant was noted after the first resection. Patients included primary (previously untreated) and previously operated or irradiated cases. Information was prospectively gathered concerning the patients' neurological condition, Karnofsky Performance Scale score, and tumor status on magnetic resonance imaging scans. There were 47 primarily operated patients (63.5%) and 27 patients (36.5%) who had previously undergone surgery or radiotherapy. A total of 121 procedures were performed in 74 patients. The mean follow-up period was 96 months, with a range of 1 to 198 months. A single stage removal was performed in 41 (55.4%) of the patients and multiple stage removal was performed in 33 (44.5%) of the patients. Gross total removal was accomplished in 53 (71.6%) of the patients, and subtotal resection was accomplished in 21 (28.4%) of the patients. During the follow-up period, 24 (32%) of the patients had no evidence of disease, 37 (50%) of the patients were alive with evidence of disease, 11 (14.8%) of the patients died of disease, and two (2.7%) of the patients died of complications. Recurrence-free survival at 10 years was 31% for the whole group, 42% for the primarily operated patients, and 26% for the reoperation cases (P = 0.0001). The average Karnofsky Performance Scale score was 80 +/- 11.7 preoperatively, 84 +/- 8.9 at the 1-year follow-up, and 86 +/- 12.8 at the last follow-up in surviving patients. No conclusion could be drawn regarding the value of radiotherapy because of the treatment philosophy and the small number of patients. Aggressive microsurgical resection of chordomas can be followed by long-term, tumor-free survival with good functional outcome. A more conservative strategy is recommended in reoperation cases, especially after previous radiotherapy, to reduce postoperative complications.


Discrepancies in carcinoembryonic antigen measurements: survey and control values vs values for patients.
We analyzed the carcinoembryonic antigen (CEA) test results reported in the College of American Pathologists' (CAP) surveys to determine the relationship between the source of CEA used to manufacture the survey specimens and the discrepancies among analytical methods. With the 1983 survey specimens, which were prepared from metastatic colon carcinoma, laboratories using Roche RIA with Clinetics columns reported results that were only one-half the values reported by laboratories using the Abbott polyclonal enzyme immunoassay. With the 1984 specimens, prepared from a different metastatic colon carcinoma, and the 1985 specimens, prepared from a tissue-culture source of CEA, the Roche results were about one-sixth as large as the Abbott results. These differences are larger than the reported assay differences for patients' specimens. In addition, twofold proportional differences were found when survey and control specimens were tested with different lots of Abbott polyclonal reagent, whereas only random differences were found with 102 patients' specimens. Evidently, assay systems perform differently with proficiency-testing and control specimens than with patients' specimens.


[Asthma and infection: risk or prevention?].
Upper respiratory tract infections and wheezing illnesses occur frequently in early childhood. Most viral infections associated with wheeze in infancy are attributable to respiratory syncitial virus, whereas in older children rhinovirus, influenza and parainfluenza virus prevail. The detection rate of viruses has increased since PCR techniques have been used. A British study for example showed that in 80% of episodes in asthmatic children aged 9-11 years viruses could be detected in nasal aspirates which had been taken within 4 days of the wheezing attack. In most cases rhinovirus was identified. Two major hypotheses have been proposed to explain the association between respiratory tract infections and subsequent respiratory abnormalities. One hypothesis states that viral infections early in life damage the growing lung or alter host immune regulation. The second hypothesis holds that respiratory infections are more severe in infants and children with some underlying predisposition. These notions await further clarification. Early childhood bacterial and viral infections may, however, also be associated with a reduced risk of developing atopic sensitization or allergic conditions, as the results of several recent studies suggest.


"Zamene," Isomeric C19 Monoolefins from Marine Zooplankton, Fishes, and Mammals.
Three isomeric C(19) monoolefins with the carbon skeleton of pristane (2,6,10,14-tetramethylpentadecane) have been isolated from mixed zooplankton of the Gulf of Maine and from liver oils of various marine fishes and mammals. The structures have been elucidated. Zamene (norphytene), the terminal olefin, is a minor component of the mixture. The usage of the older trivial name "zamene" should be restricted to the total mixture of marine C(19) monoolefins with the pristane skeleton. Conventional chemical nomenclature should be used for individual isomers.


Copy number and the stability of 2-micron circle-based artificial plasmids of Saccharomyces cerevisiae.
The copy number and stability of artificial 2-micron circle-based plasmids have been accurately measured in [Cir+] and [Cir0] strains of Saccharomyces cerevisiae. We conclude that (i) instability and copy number vary greatly from plasmid to plasmid; (ii) instability and copy number are negatively correlated--that is, high copy number is associated with low instability; (iii) it is difficult to reconcile this variability with a strict and direct system of copy number control; (iv) instabilities are much higher than expected from random partition and the observed copy numbers: this may imply partition which is less efficient than random. Even so, (v) the partitioning of 2-micron circle-like plasmids is more efficient than that of ARS-based plasmids, which hints at the existence of a system for the (inefficient) distribution of 2-micron circles.


Satisfaction with orthodontic treatment outcome.
To examine factors associated with treatment outcome satisfaction in a group of adolescent patients. One hundred and twenty patients (60 girls and 60 boys; mean age, 14.3 years; standard deviation [SD], 1.73 years) were consecutively recruited. The inclusion criteria for all patients were as follows: adolescents with a permanent dentition in need of orthodontic treatment and a treatment plan involving extractions (two or four premolars) followed by fixed appliances in both jaws. Questionnaire 1, concerning treatment motivation and expectations, was assessed prior to treatment start. Questionnaire 2 was assessed after active treatment and included questions about satisfaction with treatment outcome, quality of care and attention, and perceived pain and discomfort during active treatment. One hundred and ten patients completed the trial (54 boys and 56 girls; mean age, 16.9 years; SD, 1.78 years). Median values for satisfaction with treatment outcome were generally high. There was a clear correlation (P ≤ .001) between satisfaction with treatment outcome and patients' perception of how well they had been informed and cared for during treatment. Pain and discomfort during treatment also strongly affected treatment satisfaction. Sex, treatment time, and Peer Assessment Rating index pre- and posttreatment as well as expectations for future treatment showed no correlation with treatment satisfaction. Care and attention was the variable showing the highest correlation with satisfaction with treatment outcome. Patients' perceptions of pain and discomfort during treatment had an overall negative correlation with treatment satisfaction. Satisfaction with treatment outcome is a complex issue and requires further exploration in future research.


Impact of provider continuity on quality of care for persons with diabetes mellitus.
Many patients with diabetes fail to receive recommended monitoring tests. One reason might be inadequate continuity of care. This study examined the association between provider continuity and completion of monitoring tests for patients with diabetes mellitus. A cross-sectional analysis was conducted on claims data from a private national health plan for 1 year (January 1, 1999, through December 31, 1999). Participants had a diagnosis of diabetes mellitus and at least 2 outpatient visits during the study year (N = 1,795). The association was measured between continuity of care with an individual provider and completion of 3 diabetes monitoring tests: a glycosylated hemoglobin test, a lipid profile, and an eye examination. Eighty-one percent of patients had a glycosylated hemoglobin test, 66% had a lipid profile, and 28% had an eye examination during the study year. After controlling for demographics, number of diabetes visits, case mix, and diabetes complications, provider continuity was not significantly associated with the receipt of a glycosylated hemoglobin test (odds ratio [OR] = 0.61, 95% confidence interval [CI], 0.32-1.16), a lipid profile (OR = 0.97, 95% CI, 0.57-1.64) or an eye examination (OR = 0.60, 95% CI, 0.30-1.19). When continuity was measured only among primary care providers, there was no significant association for receipt of a glycosylated hemoglobin test (OR = 0.73, 95% CI, 0.41-1.33), a lipid profile (OR = 0.88, 95% CI, 0.53-1.47) or an eye examination (OR = 0.70, 95% CI, 0.35-1.36). This study found no association between provider continuity and completion of diabetes monitoring tests in a national privately insured population. Whereas continuity might benefit other aspects of health care, it does not appear to benefit improved monitoring for diabetes.


Lack of evidence for fecal-oral transmission of Helicobacter pylori infection in Taiwanese.
Helicobacter pylori infection is primarily acquired in early childhood and its transmission routes are debated. The aims of this study were to determine the seroprevalence of anti-H. pylori immunoglobulin G (IgG) in Taiwanese and to investigate whether a common mode of transmission could be shared between H. pylori and hepatitis A virus (HAV). An enzyme-linked immunosorbent assay (ELISA) was used to investigate the prevalence of H. pylori among 924 healthy volunteers aged less than 40 years, and radioimmunoassay for HAV infection was conducted in 500 subjects from the same population. The kappa statistic was used to measure the difference in positivity for the 2 infections in a subgroup of 500 subjects who had their sera simultaneously tested for anti-H. pylori and anti-HAV antibodies. The seroprevalence of anti-H. pylori IgG was 16.7% in subjects aged < 16 years and 38.3% at ages >or= 16 years. Most children (99%) under the age of 16 were seronegative for HAV. The prevalence of H. pylori infection increased rapidly with age, at 1%/year and 0.8%/year for children and adults, respectively. The rate of H. pylori infection was higher in male (21%) than in female (12%) children (p < 0.01; OR, 2.0; 95% confidence interval, 1.2 to 3.0). No significant difference in seroprevalence between genders was noted in subjects aged >or= 16 years. The agreement in the trend of seropositivity between both infections in the age groups 1 to 10 years and 10 to 20 years was worse than chance (kappa = -0.56) and little better than chance (kappa = 0.01), respectively. Acquisition of H. pylori infection occurs at a young age, and male children are more likely to develop the infection. The main transmission route of HAV, the fecal-oral route, did not seem to be responsible for H. pylori transmission in this Taiwanese cohort.


The Impact of SARS-CoV-2 (COVID-19) and its Lockdown Measures on the Mental and Functional Health of Older Individuals.
The effect of the COVID-19 on the physical and mental health of Italian older individuals displaying signs of cognitive deterioration has not been deeply investigated. This longitudinal study examined the impact of COVID-19 lockdown measures on the psychological well-being and motor efficiency of a sample of Italian community-dwellers with and without cognitive decline. Forty-seven participants underwent instrumental gait analysis performed in ecological setting using wearable sensors, and completed a battery of tasks assessing cognitive functioning and psychological well-being, before and after the full lockdown due to the COVID-19 spreading. A series of Multivariate Analyses of Variance (MANOVAs) documented that the superior gait performance of the cognitively healthy participants exhibited before the COVID-19 spread, vanished when they were tested at the end of the lockdown period. Moreover, before the outbreak of the COVID-19, cognitively healthy participants and those with signs of cognitive decline reported similar levels of psychological well-being, whereas, after the lockdown, the former group reported better coping, emotional competencies, and general well-being than the participants displaying signs of cognitive decline. In conclusion, the full COVID-19 outbreak had a significant impact on the mental and motor functioning of older individuals with and without signs of cognitive deterioration living in Italy.


Palatability of aposematic queen butterflies (Danaus gilippus) feeding onSarcostemma clausum (Asclepiadaceae) in Florida.
Queen butterflies (Danaus gilippus) are generally considered unpalatable to predators because they sequester and store toxic cardenolides from their larval food plants. However, a major queen food plant in Florida, the asclepiadaceous vineSarcostemma clausum, is shown here to be a very poor cardenolide source, and queens reared on this plant contain no detectable cardenolide. A direct evaluation of queen palatability using red-winged blackbirds indicates thatS. clausum-reared butterflies are essentially palatable to these predators (85% of abdomens entirely eaten), indicating little protection from either cardenolides, other sequestered phytochemicals, or de novo defensive compounds. Wild-caught queens that presumably fed as larvae uponS. clausum and also had access to adult-obtained chemicals, such as pyrrolizidine alkaloids (PAs), were relatively palatable as well (77% of abdomens eaten); they did not differ significantly in palatability from the labreared butterflies. Together, these findings suggest that; (1)S. clausumfed queens are poorly defended against some avian predators, and (2) for the particular queen sample examined, PAs do not contribute substantially to unpalatability. The discovery thatS. clausum-feeding queens are essentially palatable is of additional significance because it compels a reassessment of the classic mimicry relationship between queen and viceroy butterflies. Viceroys have been shown recently to be moderately unpalatable; therefore, the established roles of model and mimic may be reversed in some cases.


Impact of ultra-low temperature long-term storage on the preanalytical variability of twenty-one common biochemical analytes.
Retrospective studies frequently assume analytes long-term stability at ultra-low temperatures. However, these storage conditions, common among biobanks and research, may increase the preanalytical variability, adding a potential uncertainty to the measurements. This study is aimed to evaluate long-term storage stability of different analytes at <-70 °C and to assess its impact on the reference change value formula. Twenty-one analytes commonly measured in clinical laboratories were quantified in 60 serum samples. Samples were immediately aliquoted and frozen at <-70 °C, and reanalyzed after 11 ± 3.9 years of storage. A change in concentration after storage was considered relevant if the percent deviation from the baseline measurement was significant and higher than the analytical performance specifications. Preanalytical variability (CVP) due to storage, determined by the percentage deviation, showed a noticeable dispersion. Changes were relevant for alanine aminotransferase, creatinine, glucose, magnesium, potassium, sodium, total bilirubin and urate. No significant differences were found in aspartate aminotransferase, calcium, carcinoembryonic antigen, cholesterol, C-reactive protein, direct bilirubin, free thryroxine, gamma-glutamyltransferase, lactate dehydrogenase, prostate-specific antigen, triglycerides, thyrotropin, and urea. As nonnegligible, CVP must remain included in reference change value formula, which was modified to consider whether one or two samples were frozen. After long-term storage at ultra-low temperatures, there was a significant variation in some analytes that should be considered. We propose that reference change value formula should include the CVP when analyzing samples stored in these conditions.


Nocardiosis in a patient with rheumatoid arthritis treated with rituximab and a summary of reported cases.
Rituximab is a B-cell-depleting monoclonal anti-CD20 antibody. It is widely used in haematology and rheumatology. However, usage of rituximab in immunosupressed patient has been associated with various opportunistic infections. The authors reported a case of refractory rheumatoid arthritis treated with rituximab, which later presented with non-resolving pneumonia with pulmonary nodule. Percutaneous computer tomogram guided lung biopsy was arranged to confirm the suspicion of tuberculosis, but did not yield conclusive results. Later, she presented left-chest abscess and underwent incision and drainage. The pus culture and sensitivity confirmed pulmonary nocardiosis with chest wall dissemination. She was treated with 2-week course of trimethoprim sulfamethoxazole and responded. The authors also reviewed published cases of nocardiosis post-rituximab.


Cross-Sectional Association Between Blood Cell Phenotypes, Cognitive Function, and Brain Imaging Measures in the Community-Based Framingham Heart Study.
Peripheral inflammation is associated with increased risk for dementia. Neutrophil to lymphocyte ratio (NLR), red cell distribution width (RDW), and mean platelet volume (MPV), are easily measured circulating blood cell phenotypes reflecting chronic peripheral inflammation, but their association with dementia status is unclear. We sought to investigate the cross-sectional association of these inflammatory measures with neuropsychological (NP) test performance, and brain magnetic resonance imaging (MRI) measures in the Framingham Heart Study (FHS) Offspring, Third-generation, and Omni cohorts. We identified FHS participants who attended an exam that included a complete blood cell count (CBC) and underwent NP testing (n = 3,396) or brain MRI (n = 2,770) within five years of blood draw. We investigated the association between NLR, RDW, and MPV and NP test performance and structural MRI-derived volumetric measurements using linear mixed effect models accounting for family relationships and adjusting for potential confounders. Participants were on average 60 years old, 53% female, and about 80% attended some college. Higher NLR was significantly associated with poorer performance on visual memory, and visuospatial abilities, as well as with larger white matter hyperintensity volume. We also observed associations for higher RDW with poorer executive function and smaller total cerebral brain volume. Chronic peripheral inflammation as measured by NLR and RDW was associated with worse cognitive function, reduced brain volume, and greater microvascular disease in FHS participants. If confirmed in other samples, CBC may provide informative and cost-effective biomarkers of abnormal brain aging in the community.


Cartilage recovery in runners with and without knee osteoarthritis: A pilot study.
Running is an easy way of meeting physical activity recommendations for individuals with knee osteoarthritis (KOA); however, it remains unknown how their cartilage reacts to running. The objective of this pilot study was to compare the effects of 30 min of running on T2 and T1ρ relaxation times of tibiofemoral cartilage in female runners with and without KOA. Ten female runners with symptomatic KOA (mean age 52.6 ± 7.6 years) and 10 without KOA (mean age 52.5 ± 7.8 years) ran for 30 min on a treadmill. Tibiofemoral cartilage T2 and T1ρ relaxation times were measured using magnetic resonance imaging prior to and immediately after the bout of running. Repeated-measures analyses of covariance (ANCOVA) were conducted to examine between-group differences across scanning times. No Group × Time interactions were found for T2 (P ≥ 0.076) or T1ρ (P ≥ 0.288) relaxation times. However, runners with KOA showed increased T2 values compared with pre-running in the medial and lateral femur 55 min post-running (5.4 to 5.5%, P < 0.022) and in all four tibiofemoral compartments 90 min post-running (6.9 to 11.1%, P < 0.01). A significant group effect was found for T1ρ in the medial femur, with greater values in those with KOA compared with controls. While Group × Time interactions in T2 and T1ρ relaxation times remained statistically insignificant, the observed significant increases in T2 in runners with tibiofemoral osteoarthritis TFOA may suggest slower and continuing changes in the cartilage and thus a need for longer recovery after running. Future research should investigate the effects of repeated exposure to running.


Certified nurse assistants making meaning of direct care.
Using qualitative data from 87 focus groups with certified nurse assistants (CNAs) in 16 nursing homes in Massachusetts, we explore ways that CNAs make meaning of their work despite devaluations, such as lack of respect from management and residents, and the physical and emotional demands of such low-status work. CNAs' meaning making represents an effort to assert a positive identity rather than accept the stigmatization associated with their work. Assertions of the value help CNAs reconstitute their identities. Assertions of meaning, which depend upon providing good care to residents regardless of financial reward or management respect and support, make CNAs vulnerable to exploitation.


Voltage-dependent binding of dihydropyridine calcium channel blockers to guinea pig ventricular myocytes.
The voltage dependence of the binding of dihydropyridine Ca channel blockers (+)-[3H]PN200-110 and [3H]nitrendipine to enzymatically isolated guinea pig ventricular myocytes was examined. The equilibrium saturation binding of (+)-[3H]PN200-110 could be well described by a simple 1:1 binding scheme under all conditions tested. The results demonstrated that the effect of depolarization induced by high extracellular K+ (50 mM) compared to normal K+ (4.8 mM) was an increase in the observed number of binding sites (Bmax) with no change in the measured affinity of binding (Kd). Similarly, depolarization induced by a combination of Na channel toxins in the presence of normal extracellular K+ caused an increase in the observed binding of (+)-[3H]PN200-110 comparable to that observed with high K+. [3H]Nitrendipine binding was likewise increased by depolarization in the presence of 50 mM K+ compared to 4.8 mM K+. Percoll density gradient centrifugation was found to enrich the cell preparation with viable myocytes and increased the measured voltage-dependent change in binding. In agreement with the predictions from previous studies (Kamp and Miller, 1987a), the magnitude of the observed change in Bmax was directly related to the fraction of cells which were viable in a given experiment. These results are discussed in comparison to the modulated receptor hypothesis proposed from electrophysiological studies.


Plasma renin levels in Zulu hypertensives.
Plasma renin was measured by immunoassay in 10 normal Zulu controls and 72 hypertensive Zulu patients. Plasma renin values were similar in controls and patients with benign essential hypertension (1,6 plus or minus 0,83 and 1,7 plus or minus 0,21 ng/ml).


Boron excess affects photosynthesis and antioxidant apparatus of greenhouse Cucurbita pepo and Cucumis sativus.
This study aimed to evaluate the behavior of zucchini (Cucurbita pepo L.) and cucumber (Cucumis sativus L.) under boron (B) excess. Plants were grown under greenhouse conditions in a sandy soil-peat mixture using a nutrient solution containing 0.2 (control), 10 and 20 mg L(-1) B. Visible symptoms were quantified and leaf B accumulation, gas exchanges, chlorophyll (Chl) a fluorescence, malondialdehyde by-products and antioxidants were investigated 20 days after the beginning of the treatments. Boron toxicity induced oxidative load and leaf necrotic burns coupled with the reduction of leaf growth and biomass accumulation in both species. Boron excess resulted in a decrease of Chl a/b ratio, potential (Fv/Fm) and actual (ΦPSII) PSII quantum efficiency, photosynthetic rate (Pn), stomatal conductance (gs), and transpiration (E) as well. A general stimulation of the antioxidant enzymes ascorbate peroxidase, catalase and superoxide dismutase was observed, and a significant increase in the oxidized form of ascorbate and glutathione was evidenced for treated plants of both species. A difference between the two species was observed: C. pepo appeared to be more sensitive to B stress being damaged at all B concentration. C. sativus grown at 10 mg L(-1) B in nutrient solution showed some down-regulated mechanisms, i.e. increase in Chl b content and a good photochemical PSII efficiency as well as a higher amount of constitutive antioxidant molecules, that, however, are not sufficient to contrast the negative effects of B.


Sustained activation of ERK signaling in astrocytes is critical for neuronal injury-induced monocyte chemoattractant protein-1 production in rat corticostriatal slice cultures.
We previously demonstrated that N-methyl-D-aspartate (NMDA) treatment (50 microM, 3 h) induced astrocytic production of monocyte chemoattractant protein-1 (MCP-1, CCL2), a CC chemokine implicated in ischemic and excitotoxic brain injury, in rat corticostriatal slice cultures. In this study, we investigated the signaling mechanisms for NMDA-induced MCP-1 production in slice cultures. The results showed a close correlation between NMDA-induced neuronal injury and MCP-1 production, and an abrogation of NMDA-induced MCP-1 production in NMDA-pretreated slices where neuronal cells had been eliminated. These results collectively indicate that NMDA-induced neuronal injury led to astrocytic MCP-1 production. NMDA-induced MCP-1 production was significantly inhibited by U0126, an inhibitor of extracellular signal-regulated kinase (ERK). Immunostaining for phosphorylated ERK revealed that transient neuronal ERK activation was initially induced and subsided within 30 min, followed by sustained ERK activation in astrocytes. Treatment with U0126 during only the early phase (U0126 was washed out at 15 or 30 min after NMDA administration) suppressed early activation of ERK in neuronal cells, but not later activation of ERK in astrocytes. In this case, MCP-1 production was not suppressed, suggesting that activation of neuronal ERK is not necessary for MCP-1 production. In contrast, delayed application of U0126 at 3 h after the beginning of NMDA treatment inhibited MCP-1 production to the same degree as that observed when U0126 was applied from 3 h before NMDA administration. These findings suggest that sustained activation of the ERK signaling pathway in astrocytes plays a key role in neuronal injury-induced MCP-1 production.


Transcriptomic diversification of granulosa cells during follicular development between White Leghorn and Silky Fowl hens.
Egg production rate in chicken is related to the continuity of follicle development. In this study, we found that the numbers of white prehierarchical, dominant, and yellow preovulatory follicles in the high-yielding layer breed, White Leghorn (WL), were significantly higher than those in the low egg-yielding variety, Silky Fowl (SF). The proliferation and differentiation of granulosa cells (GCs) play an important role in follicle maturation. Histological observation revealed a large number of melanocytes in the outer granulosa layer of follicles in SF but not in WL. Finally, RNA-sequencing was used to analyze the gene expression profiles and pathways of the GC layer in the follicles in both WL and SF hens. Transcriptome analysis of prehierarchical GCs (phGCs) and preovulatory GCs (poGCs) between WL and SF showed that steroid hormone-, oxytocin synthesis-, tight junction-, and endocytosis-related genes were expressed at higher levels in WL phGCs than in SF phGCs, whereas the insulin signaling pathway- and vascular smooth muscle contraction-related genes were upregulated in SF phGCs. Fatty acid synthesis, calcium signaling, and Wnt signaling pathway-related genes were expressed at higher levels in WL poGCs than in SF poGCs; however, adrenergic signaling, cGMP-PKG, and melanogenesis-related genes were upregulated in SF poGCs. These results indicate that genes that promote GC proliferation and secretion of various sex hormones are more active in WL than in SF hens. The upregulated signaling pathways in SF help in providing energy to GCs and for angiogenesis and melanogenesis. In vitro experiments confirmed that both the proliferation of poGCs and synthesis of reproductive hormones were higher in WL than in SF hens.


Prostate stromal and urogenital sinus mesenchymal cell lines for investigations of stromal-epithelial interactions.
Bidirectional signaling between the urogenital sinus epithelium and mesenchyme is an essential element of prostate development that regulates ductal morphogenesis, growth, and differentiation. Comparable interactions between the epithelium and stroma in the adult prostate appear to regulate normal growth homeostasis. Alterations in the stromal-epithelial dialogue that recapitulate features of the mesenchymal-epithelial interactions of development may play a critical role in the development of benign prostatic hyperplasia and in the progression of prostate cancer. For this reason, the mesenchymal-epithelial interactions of development are of considerable interest. In this review, we provide an overview of the mesenchymal contribution to rodent prostate development with an emphasis on the stage just before ductal budding (embryonic day 16; E16) and describe the isolation, characterization and utility of a newly established E16 urogenital sinus mesenchymal cell line.


Monitoring Cell Cytoskeleton Variations upon Piezoelectric Stimulation: Implications for the Immune System.
Piezoelectric stimulation can have a significant impact on different cellular functions with possible applications in several fields, such as regenerative medicine, cancer therapy, and immunoregulation. For example, piezoelectric stimulation has been shown to modulate cytoskeleton variations: the implications of this effect range from the regulation of migration and invasion of cancer cells to the activation of pro- or anti-inflammatory phenotypes in immune cells. In this chapter, we will present different methodologies to evaluate cytoskeleton variations, focusing on modifications on f-/g-actin ratio and on the migration and invasion ability of tumor cells.


The impact of genome-wide supported schizophrenia risk variants in the neurogranin gene on brain structure and function.
The neural mechanisms underlying genetic risk for schizophrenia, a highly heritable psychiatric condition, are still under investigation. New schizophrenia risk genes discovered through genome-wide association studies (GWAS), such as neurogranin (NRGN), can be used to identify these mechanisms. In this study we examined the association of two common NRGN risk single nucleotide polymorphisms (SNPs) with functional and structural brain-based intermediate phenotypes for schizophrenia. We obtained structural, functional MRI and genotype data of 92 schizophrenia patients and 114 healthy volunteers from the multisite Mind Clinical Imaging Consortium study. Two schizophrenia-associated NRGN SNPs (rs12807809 and rs12541) were tested for association with working memory-elicited dorsolateral prefrontal cortex (DLPFC) activity and surface-wide cortical thickness. NRGN rs12541 risk allele homozygotes (TT) displayed increased working memory-related activity in several brain regions, including the left DLPFC, left insula, left somatosensory cortex and the cingulate cortex, when compared to non-risk allele carriers. NRGN rs12807809 non-risk allele (C) carriers showed reduced cortical gray matter thickness compared to risk allele homozygotes (TT) in an area comprising the right pericalcarine gyrus, the right cuneus, and the right lingual gyrus. Our study highlights the effects of schizophrenia risk variants in the NRGN gene on functional and structural brain-based intermediate phenotypes for schizophrenia. These results support recent GWAS findings and further implicate NRGN in the pathophysiology of schizophrenia by suggesting that genetic NRGN risk variants contribute to subtle changes in neural functioning and anatomy that can be quantified with neuroimaging methods.


Development and validation of a diagnostic model to differentiate spinal tuberculosis from pyogenic spondylitis by combining multiple machine learning algorithms.
This study focused on the development and validation of a diagnostic model to differentiate between spinal tuberculosis (STB) and pyogenic spondylitis (PS). We analyzed a total of 387 confirmed cases, out of which 241 were diagnosed with STB and 146 were diagnosed with PS. These cases were randomly divided into a training group (n = 271) and a validation group (n = 116). Within the training group, four machine learning (ML) algorithms (least absolute shrinkage and selection operator [LASSO], logistic regression analysis, random forest, and support vector machine recursive feature elimination [SVM-RFE]) were employed to identify distinctive variables. These specific variables were then utilized to construct a diagnostic model. The model's performance was subsequently assessed using the receiver operating characteristic (ROC) curves and the calibration curves. Finally, internal validation of the model was undertaken in the validation group. Our findings indicate that PS patients had an average platelet-to-neutrophil ratio (PNR) of 277.86, which was significantly higher than the STB patients' average of 69.88. The average age of PS patients was 54.71 years, older than the 48 years recorded for STB patients. Notably, the neutrophil-to-lymphocyte ratio (NLR) was higher in PS patients at 6.15, compared to the 3.46 NLR in STB patients. Additionally, the platelet volume distribution width (PDW) in PS patients was 0.2, compared to 0.15 in STB patients. Conversely, the mean platelet volume (MPV) was lower in PS patients at an average of 4.41, whereas STB patients averaged 8.31. Hemoglobin (HGB) levels were lower in PS patients at an average of 113.31 compared to STB patients' average of 121.64. Furthermore, the average red blood cell (RBC) count was 4.26 in PS patients, which was less than the 4.58 average observed in STB patients. After evaluation, seven key factors were identified using the four ML algorithms, forming the basis of our diagnostic model. The training and validation groups yielded area under the curve (AUC) values of 0.841 and 0.83, respectively. The calibration curves demonstrated a high alignment between the nomogram-predicted values and the actual measurements. The decision curve indicated optimal model performance with a threshold set between 2% and 88%. In conclusion, our model offers healthcare practitioners a reliable tool to efficiently and precisely differentiate between STB and PS, thereby facilitating swift and accurate diagnoses.


Coronavirus Disease 2019 Vaccine Development: An Overview.
To this day, the coronavirus disease 2019 (COVID-19) pandemic has not shown signs of abating. Moreover, the virus responsible for the pandemic, severe acute respiratory syndrome coronavirus 2, has evolved into three different variants. This phenomenon highlights an even greater need to develop drugs and vaccines to control the rate of infection and spread of the disease. As of July 7, 2020, at least 160 vaccine candidates, 21 of which have entered the clinical trial phase, have been developed. This article describes the latest advances in development, reliable platforms, strategies used, and challenges that remain in developing COVID-19 vaccines.


Second solid malignancies among children, adolescents, and young adults diagnosed with malignant bone tumors after 1976: follow-up of a Children's Oncology Group cohort.
The growing number of individuals surviving childhood cancer has increased the awareness of adverse long-term sequelae. One of the most worrisome complications after cancer therapy is the development of second malignant neoplasms (SMNs). The authors describe the incidence of solid organ SMN in survivors of pediatric malignant bone tumors who were treated on legacy Children's Cancer Group/Pediatric Oncology Group protocols from 1976 to 2005. This retrospective cohort study included 2842 patients: 1686 who were treated for osteosarcoma (OS) and 1156 who were treated for Ewing sarcoma (ES). The cohort included 56% boys/young men and 44% girls/young women, and the median age at primary diagnosis was 13 years. The median length of follow-up was 6.1 years (range, 0-20.9 years). In this analysis, 64% of patients were alive. Seventeen patients with solid organ SMN were identified. The standardized incidence ratio was 2.9 (95% confidence interval [CI], 1.4-5.4) for patients who were treated for OS and 5.0 (95% CI, 2.6-9.4) for patients who were treated for ES. The median time from diagnosis to development of solid SMN was 7 years (range, 1-13 years). The 10-year cumulative incidence of solid organ SMN for the entire cohort was 1.4% (95%CI 0.6%-2%). The magnitude of risk of solid SMNs was modest after treatment for malignant bone tumors. However, radiation-related solid SMNs will increase with longer follow-up. Because nearly 33% of patients die from their disease, recurrence remains the most significant problem. The development of improved therapies with fewer long-term consequences is paramount. Follow-up should focus on monitoring for both recurrence of primary malignancies and development of SMNs.


Insights into the Transcriptional Architecture of Behavioral Plasticity in the Honey Bee Apis mellifera.
Honey bee colonies exhibit an age-related division of labor, with worker bees performing discrete sets of behaviors throughout their lifespan. These behavioral states are associated with distinct brain transcriptomic states, yet little is known about the regulatory mechanisms governing them. We used CAGEscan (a variant of the Cap Analysis of Gene Expression technique) for the first time to characterize the promoter regions of differentially expressed brain genes during two behavioral states (brood care (aka "nursing") and foraging) and identified transcription factors (TFs) that may govern their expression. More than half of the differentially expressed TFs were associated with motifs enriched in the promoter regions of differentially expressed genes (DEGs), suggesting they are regulators of behavioral state. Strikingly, five TFs (nf-kb, egr, pax6, hairy, and clockwork orange) were predicted to co-regulate nearly half of the genes that were upregulated in foragers. Finally, differences in alternative TSS usage between nurses and foragers were detected upstream of 646 genes, whose functional analysis revealed enrichment for Gene Ontology terms associated with neural function and plasticity. This demonstrates for the first time that alternative TSSs are associated with stable differences in behavior, suggesting they may play a role in organizing behavioral state.


An appraisal of the cost benefit of magnetic resonance imaging in Nigeria.
Magnetic Resonance Scanning has revolutionized the imaging world since its invention about two decades ago. Despite its proven efficacy in patient care, it is largely non-available in the Nigeria healthcare system. This article discusses the worldwide distribution of this modality, highlights its diagnostic efficacy and emphasizes the need for Nigeria to join other countries of the world that are already utilizing this innovation in patient care. Measures at sustaining the availability of these services are suggested.


[The "best" hospitals in the Netherlands; comments on results from the Elsevier survey].
Health care workers are increasingly asked to disclose the achievements and failures of their medical interventions. Comparative evaluation of hospitals seems to be inevitable. In July 2000 about 6000 health care workers in the Netherlands received a questionnaire from the general lay weekly Elsevier asking them to grade the hospitals in their area: specialists, general practitioners, heads of departments in hospitals, nursing staff and hospital directors. The questionnaire has serious methodological flaws, e.g. regarding the items included (such as 'press sensitivity' and 'waiting lists'), the way in which the score was determined (hospitals that were scored by less than 14 respondents were excluded), the way the questions were formulated (there was no way respondents could indicate their level of experience with the hospitals involved) and the very low response rate (13%). In addition there were no data to determine the accuracy of the questionnaire, the distribution of the respondents, or whether the answers had been adjusted. The questionnaire appears to be primarily aimed at creating sensation. It received little attention in the health care sector, probably because the results were contrary to the expectations. Hospital care will undoubtedly benefit from surveys applying a limited number of well-designed indicators for quality of service, but a questionable public qualification based on a competitive model such as the Elsevier questionnaire will probably do more harm than good.


Mechanistic insights from the structures of HincII bound to cognate DNA cleaved from addition of Mg2+ and Mn2+.
The three-dimensional X-ray crystal structures of HincII bound to cognate DNA containing GTCGAC and Mn(2+) or Mg(2+), at 2.50A and 2.95A resolution, respectively, are presented. In both structures, the DNA is found cleaved, and the positions of the active-site groups, cleaved phosphate group, and 3' oxygen atom of the leaving group are in very similar positions. Two highly occupied Mn(2+) positions are found in each active site of the four crystallographically independent subunit copies in the HincII/DNA/Mn(2+) structure. The manganese ion closest to the previously identified single Ca(2+) position of HincII is shifted 1.7A and has lost direct ligation to the active-site aspartate residue, Asp127. A Mn(2+)-ligated water molecule in a position analogous to that seen in the HincII/DNA/Ca(2+) structure, and proposed to be the attacking nucleophile, is beyond hydrogen bonding distance from the active-site lysine residue, Lys129, but remains within hydrogen bonding distance from the proRp oxygen atom of the phosphate group 3' to the scissile phosphate group. In addition, the position of the cleaved phosphate group is on the opposite side of the axis connecting the two metal ions relative to that found in the BamHI/product DNA/Mn(2+) structure. Mechanistic implications are discussed, and a model for the two-metal-ion mechanism of DNA cleavage by HincII is proposed.


Focused attention improves working memory: implications for flexible-resource and discrete-capacity models.
Performance in working memory (WM) tasks depends on the capacity for storing objects and on the allocation of attention to these objects. Here, we explored how capacity models need to be augmented to account for the benefit of focusing attention on the target of recall. Participants encoded six colored disks (Experiment 1) or a set of one to eight colored disks (Experiment 2) and were cued to recall the color of a target on a color wheel. In the no-delay condition, the recall-cue was presented after a 1,000-ms retention interval, and participants could report the retrieved color immediately. In the delay condition, the recall-cue was presented at the same time as in the no-delay condition, but the opportunity to report the color was delayed. During this delay, participants could focus attention exclusively on the target. Responses deviated less from the target's color in the delay than in the no-delay condition. Mixture modeling assigned this benefit to a reduction in guessing (Experiments 1 and 2) and transposition errors (Experiment 2). We tested several computational models implementing flexible or discrete capacity allocation, aiming to explain both the effect of set size, reflecting the limited capacity of WM, and the effect of delay, reflecting the role of attention to WM representations. Both models fit the data better when a spatially graded source of transposition error is added to its assumptions. The benefits of focusing attention could be explained by allocating to this object a higher proportion of the capacity to represent color.


Frontiers of finance: evolution and efficient markets.
In this review article, we explore several recent advances in the quantitative modeling of financial markets. We begin with the Efficient Markets Hypothesis and describe how this controversial idea has stimulated a number of new directions of research, some focusing on more elaborate mathematical models that are capable of rationalizing the empirical facts, others taking a completely different tack in rejecting rationality altogether. One of the most promising directions is to view financial markets from a biological perspective and, specifically, within an evolutionary framework in which markets, instruments, institutions, and investors interact and evolve dynamically according to the "law" of economic selection. Under this view, financial agents compete and adapt, but they do not necessarily do so in an optimal fashion. Evolutionary and ecological models of financial markets is truly a new frontier whose exploration has just begun.


Effects of Three Different Orthodontic Treatment Methods on the Stability of Mandibular Incisor Alignment.
To compare the effects of extraction, non-extraction and air-rotor stripping treatments on mandibular dental arch dimensions, lower incisor positions and evaluate their effects on the stability of the treatment. The sample consisted of 44 patients with Class I malocclusion and moderate crowding including 15 patients treated with extraction, 13 with air-rotor stripping (ARS), and 16 with non-extraction treatment. The records were taken at pretreatment (T0), end of active orthodontic treatment (T1), minimum 3 years post-treatment (T2). The model and cephalometric measurements were evaluated. For statistical analyses ANOVA and Welch test was applied. At post-retention period Little's irregularity indices were increased to 1.96 mm, 2.38 mm and 3.59 mm for extraction, ARS and non-extraction groups respectively (p<0.05). At T1-T2, intercanine widths were decreased significantly at all groups (p<0.05). The arch length and arch depth decreased significantly at extraction group (p<0.05) from T0 to T1 and remained the same at T2 (p>0.05). The lower incisors were retroclined with treatment and slightly proclined at post-retention period in extraction group. In ARS and non-extraction group, lower incisors proclined with treatment and remained the same at post-retention. At all groups the irregularity indices relapsed but did not return to pretreatment values. Although significant increase at intercanine width was only observed in non-extraction treatment, at post-retention phase, intercanine widths were significantly decreased at all groups. The changes at lower incisor inclinations relapsed slightly in extraction group but remained the same in the other groups.


Alterations in the activities of hepatic plasma-membrane and microsomal enzymes during liver regeneration.
A marked increase in the activities of rat liver plasma-membrane (Na+ + K+)-stimulated ATPase and microsomal Ca2+-stimulated ATPase was observed 18h after partial hepatectomy. Lipid analyses for both membrane preparations reveal that in partially hepatectomized rats the cholesterol and sphingomyelin content are decreased with a subsequent decrease in the cholesterol/phospholipid molar ratio compared with those of sham-operated animals. Changes in the allosteric properties of plasma-membrane (Na+ + K+)-stimulated ATPase by F- (as reflected by changes in the Hill coefficient) indicated a fluidization of the lipid bilayer of both membrane preparations in 18 h-regenerating liver. The amphipathic dodecyl glucoside incorporated into the hepatic plasma membranes evoked a marked increase in the (Na+ + K+)-stimulated ATPase and 5'-nucleotidase activities. The lack of effect of the glucoside on the Lubrol-PX-solubilized 5'-nucleotidase indicates that changes in the activities of the membrane-bound enzymes caused by the glucoside are due to modulation of the membrane fluidity. Dodecyl glucoside appears to increase the membrane fluidity, evaluated through changes in the Hill coefficient for plasma-membrane (Na+ + K+)-stimulated ATPase. The biological significance of these data is discussed in terms of the differences and changes in the interaction of membrane-bound enzymes with membrane lipids during liver regeneration.


[Construction of minigenome of Newcastle disease virus of goose origin and its preliminary application].
On the base of obtaining the full length genome sequence of a Newcastle disease virus (NDV) isolated from goose, the minigenome was constructed by replacing all the encoding region with the reporter gene of enhanced green fluorescent protein (eGFP), except the virus regulating sequences relating to replication, transcription and packing of virus genome. The reporter gene could be expressed after it was transfected into the HEp-2 cells infected with helper virus of NDV. This result indicated that the minigenome could be translated by the NDV NP, P and L proteins provided by helper virus. Furthermore, the support plasmids expressing NDV NP, P and L protein were constructed respectively and the function of these plasmids was identified using the minigenome. Additionally, the virus rescue system was optimized by changing the infection dose of the recombinant vaccinia virus expressing T7 RNA polymerase. The work mentioned above will accelerate greatly the rescue of NDV and other relative research.


Molecular and functional characterization of an octopamine receptor from honeybee (Apis mellifera) brain.
Biogenic amines and their receptors regulate and modulate many physiological and behavioural processes in animals. In vertebrates, octopamine is only found in trace amounts and its function as a true neurotransmitter is unclear. In protostomes, however, octopamine can act as neurotransmitter, neuromodulator and neurohormone. In the honeybee, octopamine acts as a neuromodulator and is involved in learning and memory formation. The identification of potential octopamine receptors is decisive for an understanding of the cellular pathways involved in mediating the effects of octopamine. Here we report the cloning and functional characterization of the first octopamine receptor from the honeybee, Apis mellifera. The gene was isolated from a brain-specific cDNA library. It encodes a protein most closely related to octopamine receptors from Drosophila melanogaster and Lymnea stagnalis. Signalling properties of the cloned receptor were studied in transiently transfected human embryonic kidney (HEK) 293 cells. Nanomolar to micromolar concentrations of octopamine induced oscillatory increases in the intracellular Ca2+ concentration. In contrast to octopamine, tyramine only elicited Ca2+ responses at micromolar concentrations. The gene is abundantly expressed in many somata of the honeybee brain, suggesting that this octopamine receptor is involved in the processing of sensory inputs, antennal motor outputs and higher-order brain functions.


Exsolution of hornblende and the solubility limits of calicum in orthoamphibole.
Exsolution between orthorhombic and monoclinic amphibole has been postulated for many years on the basis of crystal-chemical and structural differences. Transmission and analytical electron microscope examination of calium-rich gedrite specimens from southwestern New Hampshire has now revealed evidence for exsolution of calcic clinoamphibole (hornblende) from ferromagnesian orthoamphibole. Analytical electron microscopy data suggests that calcium has a low solubility limit in the orthoamphibole structure. The hornblende lamellae range from only a few unit cells in thickness to about 80 nanometers. The formation of the calcic amphibole lamellae resulted from heterogeneous nucleation and growth along pre-existing (100) stacking faults. Thus, the deformation processes producing the stacking faults played a key role in preparing the sites for exsolution.


Outcomes of Combined Endocardial-Epicardial Ablation Compared With Endocardial Ablation Alone in Patients Who Undergo Epicardial Access.
Percutaneous epicardial access (EpiAcc) is used in an attempt to improve outcomes of ablation. We aim to report our experience in EpiAcc for management of symptomatic ventricular premature complexes (VPC) and ventricular tachycardia (VT). All patients from January 2004 to July 2014 who underwent EpiAcc as part of a VPC or VT ablation procedure were included. Outcomes between those with endocardial-only (Gp1) and endocardial/epicardial (Gp2) ablation and those for VPC and VT ablation were compared. EpiAcc for VPC or VT ablation was attempted in 173 patients; 10 patients were excluded because of failure of access (n = 7) or no ablation performed (n = 3). Of the remaining 163, 131 patients (80.4%) had undergone previous endocardial ablation. Mean age was 53.7 ± 15.7 years; 115 (71%) were men. VT ablation was the indication in 105 patients (64%). The underlying substrate was predominately nonischemic cardiomyopathy (49.1%). Epicardial ablation was performed in 115 (70.6%). Procedural and clinical success was obtained in 92.0% and 69.9% of patients, respectively, with no difference between Gp1 and Gp2. Those who underwent VPC ablation had superior clinical outcomes at 1-year follow-up. EpiAcc is feasible in almost all patients with no previous cardiac surgery and permits acute procedural success in >90% of patients, most of whom had failed previous ablation. However, epicardial ablation was not delivered in 1/3 of patients. Epicardial mapping may be helpful as in the absence of an appropriate epicardial site for ablation, and focus can be shifted to more detailed endocardial mapping and ablation.


Frail Older Persons' Experiences of Information and Participation in Hospital Care.
The aim of this paper is to describe frail older persons' experiences of hospital care of information and participation when being an inpatient at a hospital. A qualitative method was used. Data were collected at the hospital from 20 interviews with frail older patients, together with observations in the environment at the hospital ward. A content analysis was performed. Patients experienced not receiving information about their care and rehabilitation, or receiving such information in noisy surroundings. They experienced situations of misunderstanding related to their medication, which indicates the need for appropriate discharge calls for frail older patients. They expressed feelings of distress concerning the future, caused by hasty admissions or relatives' problems to handle the situation. The results highlight the need to receive appropriate information and to participate in decision-making. The level of health literacy should be taken notice of when giving information, using peaceful and quiet environments when informing frail older persons. Person-centered care should be recognized to a greater extent in order for healthcare professionals to give information to frail older people in a health literacy-friendly way. This might make it easier for frail older persons to participate in a partnership in care.


Fungal infections of the central nervous system: comparative analysis of risk factors and clinical signs in 57 patients.
Risk factors and clinical manifestations of fungal infections of the CNS were analyzed in 57 autopsied patients. Aspergillosis occurred in 16, candidiasis in 27, and cryptococcosis in 14. Nine of 31 variables studied showed significant difference (p less than 0.01). Cryptococcosis was community-acquired in 93%; whereas, aspergillosis and candidiasis were nosocomial in more than 95%. Focal neurologic deficits developed in 50% with CNS aspergillosis, but in only 4% with candidiasis. Meningeal signs occurred in 86% with CNS cryptococcosis, but in only 6% with aspergillosis and 7% with candidiasis. Discriminant analysis demonstrated that CNS aspergillosis was most frequently a nosocomial infection with focal neurologic deficits, pulmonary infiltrates, and hypercortisolemia. Cryptococcosis was generally a non-nosocomial infection with meningeal signs presenting in an ambulatory population. CNS candidiasis was a clinically occult nosocomial fungal infection with generally no deficits or meningismus, occurring most frequently in the neonate, the elderly, and surgical patients. The discriminant functions, which correctly classified 91% of these CNS fungal infections, may be applicable in clinical diagnosis.


[The direct AAS determination of micro elements in hair and nail by base-digestion].
The study of micro elements is more and more extensively, and people can gain some informations by the level of micro elements in tissue. This paper tempts to dissolve hair or nail in 2 mol.L-1 NaOH and determinate nine micro elements including calcium, zinc, iron, manganese, nickel, cadmium, copper, lead and bismuth in them by base-digestion with FAAS and GFAAS. It shows that the measured value of these elements is coincident with reference articles reported, except bismuth. The elements' percent recoveries are 90.0%-110.8%. The result also shows that the level of zinc and copper in hair are higher than in nail, and the level of bismuth, cadmium and iron in hair are lower than in nail, but the level of micro elements in hair and in nail are not correlative.


Langmuir monolayers of fractions of cork suberin extract.
The wide variability in composition and molecular weight of natural polymers has hampered understanding of their physicochemical properties and ultimately their use in noble applications, especially in the cases where surface properties need to be probed at the molecular level. A useful approach to analyse data from surface monolayers of complex mixtures is to try distinguishing the effects from the distinct fractions in such mixtures. The cork suberin extract investigated here is known to contain aliphatic monomers with terminal carboxylic acid and methyl ester groups as well as long esterified aliphatic chains dispersed in a polymeric aliphatic matrix. The role of such terminal groups was studied and the results showed that depending on the nature of the terminal groups the monolayers present distinct isotherms due to the different interactions with the water subphase. Fractionation strategies based on different solubilities of the cork suberin components in chloroform were also employed to probe their effect on the monolayer characteristics. From the two sets of experiments it is clear that the presence of monomers with terminal carboxylic acids in the suberin extract affects considerably the monolayer-forming ability. This approach may be used as a complementary, relatively simple route to assess suberin genetic engineering strategies towards resistance to environmental stress.


Noncanonical regulation of phosphatidylserine metabolism by a Sec14-like protein and a lipid kinase.
The yeast phosphatidylserine (PtdSer) decarboxylase Psd2 is proposed to engage in a membrane contact site (MCS) for PtdSer decarboxylation to phosphatidylethanolamine (PtdEtn). This proposed MCS harbors Psd2, the Sec14-like phosphatidylinositol transfer protein (PITP) Sfh4, the Stt4 phosphatidylinositol (PtdIns) 4-OH kinase, the Scs2 tether, and an uncharacterized protein. We report that, of these components, only Sfh4 and Stt4 regulate Psd2 activity in vivo. They do so via distinct mechanisms. Sfh4 operates via a mechanism for which its PtdIns-transfer activity is dispensable but requires an Sfh4-Psd2 physical interaction. The other requires Stt4-mediated production of PtdIns-4-phosphate (PtdIns4P), where Stt4 (along with the Sac1 PtdIns4P phosphatase and endoplasmic reticulum-plasma membrane tethers) indirectly modulate Psd2 activity via a PtdIns4P homeostatic mechanism that influences PtdSer accessibility to Psd2. These results identify an example in which the biological function of a Sec14-like PITP is cleanly uncoupled from its canonical in vitro PtdIns-transfer activity and challenge popular functional assumptions regarding lipid-transfer protein involvements in MCS function.


[The role of Ca2+ ions in a phage infection of Corynebacterium glutamicum].
It was shown that neither uncouplers of oxidative phosphorylation, nor lack of Ca2+ ions affected the normal MC-2 phage absorption on Corynebacterium glutamicum cells, while the phage development was repressed under these conditions. Simultaneous measurement of Ca2+, K+ and H+ ion flows, as well as measurement of membrane potential showed that the addition of the phage into the experimental medium led to significant depolarization of the membrane from -160 mV to -100 mV due to the penetration of Ca2+ ions into the cells followed by K+ and H+ efflux. The (Ca2+) to (K+ + H+) ratio was shown to be 1 : 1. Phage DNA is supposed to be injected into the host cells as a positively charged (Ca2+-DNA) complex.


Transient upregulation of CBFA1 in response to bone morphogenetic protein-2 and transforming growth factor beta1 in C2C12 myogenic cells coincides with suppression of the myogenic phenotype but is not sufficient for osteoblast differentiation.
The bone morphogenetic protein (BMP)-2 is a potent osteoinductive signal, inducing bone formation in vivo and osteoblast differentiation from non-osseous cells in vitro. The runt domain-related protein Cbfa1/PEBP2alphaA/AML-3 is a critical component of bone formation in vivo and transcriptional regulator of osteoblast differentiation. To investigate the relationship between the extracellular BMP-2 signal, Cbfa1, and osteogenesis, we examined expression of Cbfa1 and osteoblastic genes during the BMP-2 induced osteogenic transdifferentiation of the myoblastic cell line C2C12. BMP-2 treatment completely blocked myotube formation and transiently induced expression of Cbfa1 and the bone-related homeodomain protein Msx-2 concomitant with loss of the myoblast phenotype. While induction of collagen type I and alkaline phosphatase (AP) expression coincided with Cbfa1 expression, Cbfa1 mRNA was strikingly downregulated at the onset of expression of osteopontin (OPN) and osteocalcin (OCN) genes, reflecting the mature osteoblast phenotype. TGF-beta1 treatment effectively suppressed myogenesis and induced Cbfa1 expression but was insufficient to support osteoblast differentiation reflected by the absence of ALP, OPN, and OCN. We addressed whether induction of Cbfa1 in response to BMP-2 results in the transcriptional activation of the OC promoter which contains three enhancer Cbfa1 elements. Transfection studies show BMP-2 suppresses OC promoter activity in C2C12, but not in osteoblastic ROS 17/2.8 cells. Maximal suppression of OC promoter activity in response to BMP-2 requires sequences in the proximal promoter (up to nt -365) and may occur independent of the three Cbfa sites. Taken together, our results demonstrate a dissociation of Cbfa1 expression from development of the osteoblast phenotype. Our findings suggest that Cbfal may function transiently to divert a committed myoblast to a potentially osteogenic cell. However, other factors induced by BMP-2 appear to be necessary for complete expression of the osteoblast phenotype.


Superior Colliculus Lesions Lead to Disrupted Responses to Light in Diurnal Grass Rats (Arvicanthis niloticus).
The circadian system regulates daily rhythms of physiology and behavior. Although extraordinary advances have been made to elucidate the brain mechanisms underlying the circadian system in nocturnal species, less is known in diurnal species. Recent studies have shown that retinorecipient brain areas such as the intergeniculate leaflet (IGL) and olivary pretectal nucleus (OPT) are critical for the display of normal patterns of daily activity in diurnal grass rats (Arvicanthis niloticus). Specifically, grass rats with IGL and OPT lesions respond to light in similar ways to intact nocturnal animals. Importantly, both the IGL and OPT project to one another in nocturnal species, and there is evidence that these 2 brain regions also project to the superior colliculus (SC). The SC receives direct retinal input, is involved in the triggering of rapid eye movement sleep in nocturnal rats, and is disproportionately large in the diurnal grass rat. The objective of the current study was to use diurnal grass rats to test the hypothesis that the SC is critical for the expression of diurnal behavior and physiology. We performed bilateral electrolytic lesions of the SC in female grass rats to examine behavioral patterns and acute responses to light. Most grass rats with SC lesions expressed significantly reduced activity in the presence of light. Exposing these grass rats to constant darkness reinstated activity levels during the subjective day, suggesting that light masks their ability to display a diurnal activity profile in 12:12 LD. Altogether, our data suggest that the SC is critical for maintaining normal responses to light in female grass rats.


Revising the Roles of Aldosterone in Vascular Physiology and Pathophysiology: From Electocortin to Baxdrostat.
Aldosterone was initially identified as a hormone primarily related to regulation of fluid and electrolyte homeostasis. However, over the past 20 years there has been an increasing appreciation of its role in regulation of vascular function and pathophysiology in the setting of hypertension, atherosclerosis, and heart failure. This review highlights recent advances in our understanding the biology of aldosterone as it relates to the pathophysiology and the management of vascular disease-especially related to hypertension. The review focuses on 3 key areas: 1) advances in our understanding of the cellular mechanisms by which aldosterone mediates its cellular effects, 2) identification of the hidden epidemic of aldosteronism as a mediator of hypertension, and 3) appreciating new therapeutic advances in the clinical pharmacology of aldosterone inhibition in cardiovascular and renal disease.


Autonomic modulation in hypertension without hypertrophy.
Cardiac autonomic modulation and baroreflex sensitivity are altered in individuals with essential hypertension. Hypertension is considered as a strong and independent risk factor for supraventricular and ventricular arrhythmias. The aim of the present study was to evaluate cardiac autonomic control and the arrhythmogenic risk by using 24-h heart rate variability (HRV) and heart rate turbulence (HRT) analysis in essential hypertension without left ventricular hypertrophy (LVH). Fifty-eight newly diagnosed untreated hypertensive patients without LVH (mean age 51 ± 12 years, 26 women) and 56 adult, healthy volunteers (mean age 49 ± 12 years, 24 women) were included in the study. Subjects with secondary causes of hypertension or autonomic dysfunction were excluded. The diagnosis of hypertension was based on ambulatory blood pressure monitoring results. Time-domain HRV parameters and HRT parameters were calculated from 24-hour Holter recordings. Values of SDNN and SDANN in the hypertension group were significantly lower than in the control group (P < 0.01 and P < 0.01, respectively). At least one of the turbulence onset (TO) and turbulence slope (TS) values were found abnormal in 10 of 32 hypertensive patients and in 5 of 24 control individuals (P = 0.38). There was no significant difference between HRT parameters, TO and TS (P = 0.67 and P = 0.12, respectively). Sympathetic tone begins to increase in hypertension before LVH develops. However, the impact of this increase on HRT is not clear. There is a need for further research to investigate the impact of hypertension and LVH on HRT.


Impact of supplementation with a food-derived microbial community on obesity-associated inflammation and gut microbiota composition.
Obesity is a complex pathology associated with dysbiosis, metabolic alterations, and low-grade chronic inflammation promoted by immune cells, infiltrating and populating the adipose tissue. Probiotic supplementation was suggested to be capable of counteracting obesity-associated immune and microbial alterations, based on its proven immunomodulatory activity and positive effect on gut microbial balance. Traditional fermented foods represent a natural source of live microbes, including environmental strains with probiotic features, which could transiently colonise the gut. The aim of our work was to evaluate the impact of supplementation with a complex foodborne bacterial consortium on obesity-associated inflammation and gut microbiota composition in a mouse model. C57BL/6J mice fed a 45% high fat diet (HFD) for 90 days were supplemented with a mixture of foodborne lactic acid bacteria derived from the traditional fermented dairy product "Mozzarella di Bufala Campana" (MBC) or with the commercial probiotic GG strain of Lactobacillus rhamnosus (LGG). Inflammation was assessed in epididymal white adipose tissue (WAT) following HFD. Faecal microbiota composition was studied by next-generation sequencing. Significant reduction of epididymal WAT weight was observed in MBC-treated, as compared to LGG and control, animals. Serum metabolic profiling showed correspondingly reduced levels of triglycerides and higher levels of HDL cholesterol, as well as a trend toward reduction of LDL-cholesterol levels. Analysis of the principal leucocyte subpopulations in epididymal WAT revealed increased regulatory T cells and CD4+ cells in MBC microbiota-supplemented mice, as well as decreased macrophage and CD8+ cell numbers, suggesting anti-inflammatory effects. These results were associated with lower levels of pro-inflammatory cytokines and chemokines in WAT explants. Faecal bacterial profiling demonstrated increased Firmicutes/Bacteroidetes ratio in all mice groups following HFD. Taken together, these results indicate a protective effect of MBC microbiota supplementation toward HFD-induced fat accumulation and triglyceride and cholesterol levels, as well as inflammation, suggesting a stronger effect of a mixed microbial consortium vs single-strain probiotic supplementation. The immunomodulatory activity exerted by the MBC microbiota could be due to synergistic interactions within the microbial consortium, highlighting the important role of dietary microbes with yet uncharacterised probiotic effect.


The lncRNA NORAD/miR-520a-3p Facilitates Malignancy in Non-Small Cell Lung Cancer via PI3k/Akt/mTOR Signaling Pathway.
The effects of lncRNA-NORAD/mir-520a-3p on proliferation and invasion of non-small cell lung cancer (NSCLC) were studied, and its potential molecular mechanism was discussed. qRT-PCR was used to detect the expression of lncRNA NORAD and miR-520a-3p in non-small cell lung cancer tissues and cell lines. CCK-8 method and Transwell test were used to identify the effects of lncRNA NORAD on the proliferation and invasion in NSCLC. Target gene prediction and screening and luciferase reporter assay was used to verify downstream target genes of lncRNA NORAD. The expressions of PI3K, AKT, and mTOR proteins were detected by Western blot. Compared with normal tissues and cells, the expressions of lncRNA NORAD in cancer tissues and cells were significantly higher. Compared with normal cells, the expression of miR-520a-3p in cells was considerably lower. LncRNA NORAD could accelerate the growth and metastasis of NSCLC in vitro and in vivo. Luciferase reporter assay results indicated that miR-520a-3p was a downstream target gene of lncRNA NORAD. Further findings showed that lncRNA NORAD might bind to miR-520a-3p, thereby affecting the PI3k/Akt/mTOR signaling pathway. LncRNA NORAD can regulate the proliferation of NSCLC by regulating miR-520a-3p/PI3k/Akt/mTOR signaling pathway, thus promoting the occurrence and development of NSCLC.


Aqueous Phase Separation of Responsive Copolymers for Sustainable and Mechanically Stable Membranes.
Membranes are often used in environmentally friendly applications and as a sustainable alternative to conventional processes. Unfortunately, the vast majority of polymeric membranes are produced via an unsustainable and environmentally unfriendly process that requires large amounts of harsh reprotoxic chemicals such as N-methyl-2-pyrrolidinone and dimethylformamide. In this work, we investigate an aqueous phase separation (APS) system that uses weak polyelectrolytes, whose charge is dependent on the pH (weak polyelectrolytes), to produce membranes. Specifically the copolymer polystyrene-alt-maleic acid (PSaMA) is used. PSaMA contains responsive monomers, required for APS, and also unresponsive hydrophobic monomers that provide mechanical stability to the resultant membranes. This work demonstrates that by controlling the precipitation of PSaMA, it is possible to prepare a wide range of membranes; from microfiltration membranes capable of treating oily waste water to dense nanofiltration-type membranes with excellent micropollutant retentions and high mechanical stability. While similar materials in prior work could only withstand 4 bar, the membranes presented here demonstrate stable operation up to 20 bar. The only solvents used in this APS system are water and the green solvent acetic acid, thus making our APS process significantly more sustainable and environmentally friendly as compared to the conventional membrane fabrication methods.


Correlated evolution of allometry and sexual dimorphism across higher taxa.
Empirical evidence suggests that Rensch's rule of allometric scaling of male and female body size, which states that body size divergence is greater across males than across females of a clade, is not universal. In fact, quantitative genetic theory indicates that the sex under historically stronger directional selection will exhibit greater interspecific variance in size. Thus, the pattern of covariance between allometry of male and female body size and sexual size dimorphism (SSD) across related clades allows a test of this causal hypothesis for macroevolutionary trends in SSD. We compiled a data set of published body size estimates from the amphibians, a class with predominantly female-biased SSD, to examine variation in allometry and SSD among clades. Our results indicate that females become the more size-variant sex across species in a family as the magnitude of SSD in that family increases. This rejects Rensch's rule and implicates selection on females as a driver of both amphibian allometry and SSD. Further, when we combine our data into a single analysis of allometry for the class, we find a significant nonlinear allometric relationship between female body size and male body size. These data suggest that allometry changes significantly as a function of size. Our results illustrate that the relationship between female size and male size varies with both the degree of sexual dimorphism and the body size of a clade.


Evaluation of the distance between Schwalbe's line and the anterior lens capsule as a parameter for the correction of ultrasound biomicroscopic values of the canine iridocorneal angle.
To determine whether the distance between Schwalbe's line (the peripheral termination of Descemet's membrane, i.e., the borderline between the cornea and sclera) and the anterior lens capsule (SLD) is an applicable parameter for correcting raw ultrasound biomicroscopic values of the canine iridocorneal angle (ICA) and to establish a comparative system for the ICA values in different canine breeds with varied body sizes/weights. The dogs were divided into four groups based on body weight (BW): <4 kg, 4-8 kg, 8-20 kg, and >20 kg, and 180 normotensive eyes were studied. The ICA microstructure was examined by ultrasound biomicroscopy (UBM) using 40-MHz probes in dogs with or without anesthesia/sedation. Linear regression analysis and correlation coefficients were evaluated between SLD or SLD(2) and UBM measurements; subsequently, noncorrected and SLD-corrected UBM values were statistically assessed. Significant linear correlations were detected between SLD and the ciliary cleft width, the minimum distance between the angle recess and the scleral venous plexus, and the scleral thickness. Positive linear correlations were also demonstrated between SLD(2) and the ciliary cleft area as well as the scleral venous plexus area. Raw UBM measurements were corrected by using a ratio with SLD on the distance or SLD(2) on the area. Although noncorrected UBM measurements increased with canine BW, SLD-corrected UBM values remained similar with no significant statistical differences in any of the dogs. Correction with SLD would be clinically useful for comparing UBM measurements of the ICA in dogs with different body sizes/weights.


Incremental cost of nosocomial bacteremia according to the focus of infection and antibiotic sensitivity of the causative microorganism in a university hospital.
To estimate the incremental cost of nosocomial bacteremia according to the causative focus and classified by the antibiotic sensitivity of the microorganism.Patients admitted to Hospital del Mar in Barcelona from 2005 to 2012 were included. We analyzed the total hospital costs of patients with nosocomial bacteremia caused by microorganisms with a high prevalence and, often, with multidrug-resistance. A control group was defined by selecting patients without bacteremia in the same diagnosis-related group.Our hospital has a cost accounting system (full-costing) that uses activity-based criteria to estimate per-patient costs. A logistic regression was fitted to estimate the probability of developing bacteremia (propensity score) and was used for propensity-score matching adjustment. This propensity score was included in an econometric model to adjust the incremental cost of patients with bacteremia with differentiation of the causative focus and antibiotic sensitivity.The mean incremental cost was estimated at &OV0556;15,526. The lowest incremental cost corresponded to bacteremia caused by multidrug-sensitive urinary infection (&OV0556;6786) and the highest to primary or unknown sources of bacteremia caused by multidrug-resistant microorganisms (&OV0556;29,186).This is one of the first analyses to include all episodes of bacteremia produced during hospital stays in a single study. The study included accurate information about the focus and antibiotic sensitivity of the causative organism and actual hospital costs. It provides information that could be useful to improve, establish, and prioritize prevention strategies for nosocomial infections.


Dissemination of transferable CTX-M-type extended-spectrum beta-lactamase-producing Escherichia coli in Korea.
Among 365 Escherichia coli isolated in 2003, 31 cefotaxime-resistant isolates were obtained from clinical specimens taken from adults hospitalized in Busan, Korea. Six extended-spectrum beta-lactamase (ESBL)-producing isolates were investigated further to determine the mechanism of resistance. These isolates were analysed by antibiotic susceptibility testing, pI determination, plasmid profiles, transconjugation test, PCR-restriction fragment length polymorphism (RFLP), enterobacterial repetitive consensus (ERIC)-PCR and DNA sequencing. All six of these isolates were found to contain the CTX-M-type ESBL genes. Five clinical isolates and their transconjugants produced CTX-M-3. One clinical isolate (K17391) and its transconjugant (trcK17391) produced CTX-M-15. Five clinical isolates also produced another TEM-1. One clinical isolate (K12776) also contained another TEM-52. CTX-M-3 ESBL gene was responsible for the resistance to piperacillin, cephalothin, cefotaxime, cefepime and aztreonam. CTX-M-15 or TEM-52 was especially responsible for the resistance to ceftazidime. These results appear to represent the in vivo evolution of CTX-M-type beta-lactamase genes (bla(CTX-M-3) --> bla(CTX-M-15)) under the selective pressure of antimicrobial therapy (especially ceftazidime). PCR-RFLP is a reliable method to discriminate CTX-M-15 gene from CTX-M-3 gene. ERIC-PCR analysis revealed that dissemination of CTX-M-3 was not due to a clonal outbreak of a resistant strain but to the intra-species spread of resistance to piperacillin, cephalothin, cefotaxime, cefepime and aztreonam in Korea. This is the first report of the occurrence of CTX-M-1 cluster ESBLs in Korea. A more comprehensive survey of these ESBL types from Korea is urgently needed because of the in vivo evolution of CTX-M-15 from CTX-M-3. The emergence of these CTX-M-type ESBLs suggests that diagnostic laboratories should screen for ESBLs with ceftazidime as well as cefotaxime; they should still perform clavulanate synergy tests on resistant isolates.


The palliative index: predicting outcomes of emergent surgery in patients with cancer.
The role of emergent palliative surgery in the setting of advanced malignancy remains a subject of controversy. The purpose of this study was to identify clinical predictors of outcome in patients with cancer who undergo nonelective abdominal surgery. Individuals who underwent urgent and emergent abdominal operations between 2006 and 2010 at a tertiary cancer center were identified. Analyses were performed to identify predictors of 30-day morbidity and mortality as well as overall survival (OS). A risk score was derived from predictors of OS. Of 143 patients, 93 (65%) had active disease (AD; defined as evidence of malignancy at time of surgery). Thirty-day morbidity and mortality were 36.4% and 9.8%, respectively. Independent predictors of 30-day mortality included ASA score >3 (p=0.009) and albumin <2.8 (p=0.040). Median OS was 5.4 months in patients with AD and was not reached in patients without AD (p<0.001). Independent predictors of decreased OS included AD; ASA >3; creatinine >1.3; and a tumor-related indication (i.e., bleeding, obstructing, or perforating tumor). A risk or palliative index (PI) score stratified patients into groups with discreet outcomes. Although AD did not predict 30-day morbidity, it was the dominant independent predictor of postoperative OS. In cancer patients undergoing emergency abdominal surgery, outcome is anticipated by disease status and other independent predictors of OS.


Isolated hepatic tuberculosis: a rare cause of hepatic mass lesions.
Hepatic tuberculosis usually accompanies pulmonary and extrapulmonary tuberculosis. Although isolated hepatic tuberculosis is a very rare condition, it should be considered in the differential diagnosis of a hepatic mass. Here, we report a 42-year-old woman presenting with weight loss, fever, night sweats, and a hepatic mass on the abdominal ultrasonography and magnetic resonance imaging (MRI). Ultrasonography-guided percutaneous needle biopsy demonstrated a caseating granuloma with epithelioid histiocytes and giant cells compatible with the diagnosis of tuberculosis. The patient was treated with four anti-tuberculous drugs for 1 year. She recovered clinically, and her post-treatment abdominal MRI was normal.


Expression and Purification of Delta Sleep-Inducing Peptide Fused with Protein Transduction Domain and Human Serum Albumin in Pichia pastoris.
Sleep is a natural part of every individual's life. Delta sleep-inducing peptide (DSIP) is a nonapeptide that could promote sleep through the induction of slow wave sleep. However, little is known about the pharmacological effect of DSIP on insomnia. The main objective of this study was to analyze the pharmacological effect of DSIP on insomnia. We designed a fusion protein containing N-terminal TAT-based transduction domain followed by human serum albumin and DSIP and designated this protein as PHD fusion protein. The PHD fusion protein were expressed in Pichia pastoris and purified. Mice were administered single subcutaneous injections three concentrations of PHD fusion protein (0.5, 1, 2 mg/kg), and the pharmacological activity of PHD fusion protein was studied using classic pentobarbitalinduced sleep test. We expressed the PHD fusion protein in P. pastoris; furthermore, the PHD fused protein was purified to near homogeneity by DEAE Sepharose FF, Phenyl Sepharose HP and Blue Sepharose 6 FF. Our result showed that the increase of pentobarbital-induced hypnotic effect characterized by reducing sleep latency and prolonged sleep duration was observed for increasing concentrations of PHD fusion protein (P<0.05); moreover, different dose of PHD fusion protein could induce the mice to re-sleep in a dose-dependent manner, whereas higher doses of PHD fusion protein (1.0, 2.0 mg/kg) significantly increased the rate of sleep re-onset compared with the vehicle group of mice (P<0.05). PHD fusion protein increased the hypnotic effects of pentobarbital by reducing sleep latency and prolonged sleep duration. The present study suggested PHD fusion protein could be a new drug candidate for insomnia.


Serum Tamm-Horsfall glycoprotein levels in health and in renal disease.
Serum Tamm-Horsfall glycoprotein (THG) concentrations were measured by radioimmunoassay in a group of 95 people with normal renal function and lay in the range of 70 to 540 ng/ml. None was detectable in 6 anephric patients. Patients on hemodialysis but with both kidneys in situ had concentrations below the lower limit of normal, and in some cases THG was undetectable. Healthy kidney donors generally showed a decrease in serum levels following uninephrectomy although these remained within the normal range. In contrast, all transplant recipients showed a marked increase in serum THG concentration, the rates of rise being apparently dependent upon initial graft function. These findings support the view that THG is a normal constituent of human serum whose levels are related to the amount of renal functioning mass.


S-nitrosothiols modulate G protein-coupled receptor signaling in a reversible and highly receptor-specific manner.
Recent studies indicate that the G protein-coupled receptor (GPCR) signaling machinery can serve as a direct target of reactive oxygen species, including nitric oxide (NO) and S-nitrosothiols (RSNOs). To gain a broader view into the way that receptor-dependent G protein activation -- an early step in signal transduction -- might be affected by RSNOs, we have studied several receptors coupling to the Gi family of G proteins in their native cellular environment using the powerful functional approach of [35S]GTPgammaS autoradiography with brain cryostat sections in combination with classical G protein activation assays. We demonstrate that RSNOs, like S-nitrosoglutathione (GSNO) and S-nitrosocysteine (CysNO), can modulate GPCR signaling via reversible, thiol-sensitive mechanisms probably involving S-nitrosylation. RSNOs are capable of very targeted regulation, as they potentiate the signaling of some receptors (exemplified by the M2/M4 muscarinic cholinergic receptors), inhibit others (P2Y12 purinergic, LPA1lysophosphatidic acid, and cannabinoid CB1 receptors), but may only marginally affect signaling of others, such as adenosine A1, mu-opioid, and opiate related receptors. Amplification of M2/M4 muscarinic responses is explained by an accelerated rate of guanine nucleotide exchange, as well as an increased number of high-affinity [35S]GTPgammaS binding sites available for the agonist-activated receptor. GSNO amplified human M4 receptor signaling also under heterologous expression in CHO cells, but the effect diminished with increasing constitutive receptor activity. RSNOs markedly inhibited P2Y12 receptor signaling in native tissues (rat brain and human platelets), but failed to affect human P2Y12 receptor signaling under heterologous expression in CHO cells, indicating that the native cellular signaling partners, rather than the P2Y12 receptor protein, act as a molecular target for this action. These in vitro studies show for the first time in a broader general context that RSNOs are capable of modulating GPCR signaling in a reversible and highly receptor-specific manner. Given that the enzymatic machinery responsible for endogenous NO production is located in close proximity with the GPCR signaling complex, especially with that for several receptors whose signaling is shown here to be modulated by exogenous RSNOs, our data suggest that GPCR signaling in vivo is likely to be subject to substantial, and highly receptor-specific modulation by NO-derived RSNOs.


[Nutrition disorders in persons qualified for the methadone treatment. Part II. Food choice and intake in diets of opiate addicts].
The aim of the study is to establish the composition and nutritional value of daily food in the group of opiate addicted patients qualified to methadone therapy. The presented data show the results of the survey carried out at the start of the treatment in the group of 42 patients (13 women aged from 21-33 years and 29 men aged from 21-46 years). For the dietary survey the method of 24 hour recall was used: nutrients of daily individual diets and mean values with standard deviations for examined groups were calculated with use of own computer programme and polish food composition data. The proportion of energy covered by groups of food products was also calculated. The results show wide distribution of food consumption: the mean level of the daily energy intake was 2438 kcal (SD +/- 976, range 492 kcal to 4193 kcal.) in the group of men, and 2039 kcal (SD +/- 925, range 561-3860 kcal) in the group of women. The mean protein intake was 82 g (13.4 E%) for men and 63 g (12.4 E%) for women. In the diet of 5 men and 6 women carbohydrate energy exceeded 70% of the total energy intake, indicating high consumption of food items containing considerable amounts of sugar (bonbons, confectionery, cakes, sweets, chocolate, sweet drinks and similar).


Postnatal developmental changes in AMPA and NMDA receptors in the rat vestibular nuclei.
Changes in the expression of the AMPA receptor subunits GluR1-4 and of the NMDA receptor subunits NR1, NR2A-D were investigated in the developing rat medial and lateral vestibular nuclei. Analyses were performed using nonradioactive in situ hybridization and immunoblotting with subunit-specific antibodies. During the postnatal development, glutamatergic receptor subunits were differentially expressed in the vestibular nuclei. The level of expression of GluR1, GluR4 and NR1 subunits was higher in the developing brain as compared to the adult. We observed a gradual increase in GluR2/3, NR2A, NR2B and NR2C levels of expression in the medial and lateral vestibular nuclei during the first 3 weeks of postnatal development. In situ hybridization results were consistent with immunoblot analyses. The differential expression of AMPA and NMDA receptor subunits in immature vestibular neurons is consistent with changes in glutamate receptor properties. This may be related to the postsynaptic regulation of receptor subunits associated with the synaptic plasticity of the vestibular neuron connections during specific sequences of postnatal development.


It is all about proteases: from drug delivery to in vivo imaging and photomedicine.
Clinical studies provide overwhelming evidence for the importance of proteolytic imbalance and the upregulation of diverse protease classes in diseases such as cancer and arthritis. While the complex nature of proteolytic networks has hampered the development of protease inhibitors for these indications, aberrant enzyme activity could be successfully exploited for the development of proteasesensitive drug delivery systems and fluorescent in vivo imaging agents. More recently, these concepts have also been translated into photomedical applications to develop dual modality prodrugs for the simultaneous treatment and imaging of disease. After an introductory overview of proteases and their role in cancer, we present and discuss different strategies to exploit upregulated protease activity for the development of drug delivery systems, fluorescent in vivo reporter probes, and photosensitizer-prodrugs with respect to their potential and limitations. The main approaches used for targeting proteases in all three areas can be roughly divided into peptide-based and macromolecular strategies. Both involve the use of a short, peptide-based protease substrate, which is either directly tagged to the therapeutic agent or dye/quencher pair, or alternatively, serves as a linker between the polymeric carrier and a functional unit. In the latter case, the pharmacokinetic properties of peptide-based protease-sensitive prodrugs and imaging probes can be further ameliorated by the passive targeting capacity of macromolecular drug delivery systems for neoplastic and inflammatory lesions.


An optical counting technique with vertical hydrodynamic focusing for biological cells.
A BARRIER IN SCALING LABORATORY PROCESSES INTO AUTOMATED MICROFLUIDIC DEVICES HAS BEEN THE TRANSFER OF LABORATORY BASED ASSAYS: Where engineering meets biological protocol. One basic requirement is to reliably and accurately know the distribution and number of biological cells being dispensed. In this study, a novel optical counting technique to efficiently quantify the number of cells flowing into a microtube is presented. REH, B-lymphoid precursor leukemia, are stained with a fluorescent dye and frames of moving cells are recorded using a charge coupled device (CCD) camera. The basic principle is to calculate the total fluorescence intensity of the image and to divide it by the average intensity of a single cell. This method allows counting the number of cells with an uncertainty +/-5%, which compares favorably to the standard biological methodology, based on the manual Trypan Blue assay, which is destructive to the cells and presents an uncertainty in the order of 20%. The use of a microdevice for vertical hydrodynamic focusing, which can reduce the background noise of out of focus cells by concentrating the cells in a thin layer, has further improved the technique. Computational fluid dynamics (CFD) simulation and confocal laser scanning microscopy images have shown an 82% reduction in the vertical displacement of the cells. For the flow rates imposed during this study, a throughput of 100-200 cellss is achieved.


Nonrandom distribution of repeated DNA sequences with respect to supercoiled loops and the nuclear matrix.
The DNA in a eukaryotic nucleus is arranged into a series of supercoiled loops that are anchored at their bases to the nuclear matrix. We have analyzed the DNA sequences that are closest to the matrix attachment points for their relative content of specific repeated sequences. Sequences were enriched (mouse satellite, human Alu family) or depleted (mouse EcoRI repeat, monkey alpha component), depending on the specific sequence and species examined. These results can be understood in terms of a nonrandom arrangement of DNA sequences with respect to nuclear DNA loops.