Serum VEGF and CFH in exudative age-related macular degeneration.
To determine serum vascular endothelial growth factor 165 (VEGF165) levels and the association of the complement factor H gene (CFH) Y402H polymorphism in patients with exudative age-related macular degeneration (AMD) in comparison to unaffected control subjects. Sixty-six AMD patients and 66 healthy age- and gender-matched controls were included in this case-control study. The serum VEGF165 was assayed by ELISA (R&D). Genotypes were determined by polymerase chain reaction-restriction fragment length polymorphism analysis. Chi-squared tests were used regarding the polymorphism, a t-test regarding the VEGF-levels. Levels of serum VEGF165 were similar in both groups (p-value = 0.2112). Genotype frequency differed significantly between patients with exudative AMD and the healthy control group (p = 0.003136). The serum VEGF165 levels were similar irrespective of the presence of the CFH Y402H polymorphism (p = 0.4113) and independent of the specific genotype (p = 0.9634). In the present study, exudative AMD is not associated to serum VEGF165 levels; furthermore, our data does not establish a statistical link between VEGF165 and the CFH Y402H polymorphism.


Can Brain Natriuretic Peptides and Osteoprotegerin Serve As Biochemical Markers for the Detection of Aortic Pathology in Children and Adolescents with Turner Syndrome?
Turner syndrome (TS) is a chromosomal disorder that affects 1:2,000 females. It results from either the complete or partial loss of the X chromosome as well as other aberrations. Clinical features of TS include short stature, delayed puberty, and congenital cardiac malformations. TS children also have an increased prevalence of cardiometabolic risk factors, which predisposes them to complications like coronary artery disease, cerebrovascular-related deaths, and aortic dissection. Early cardiac imaging, such as echocardiography and cardiac magnetic resonance imaging, are recommended to detect underlying aortic pathology. However, these modalities are limited by cost, accessibility, and are operator dependent. In view of these shortcomings, alternative methods, like vascular biomarkers, are currently being explored. There are only a few studies that have examined the relationship between B-type natriuretic peptide (BNP), N-terminal pro BNP (NT pro-BNP), and osteoprotegerin (OPG) and aortic disease in TS, and thus the data are only in proof-of-concept stages. Further meticulous longitudinal studies are required before BNP, NT pro-BNP, and OPG are used as vascular biomarkers for the detection of aortic disease in childhood and adolescent TS.


[A method for the differential staining of muscle and connective tissues from organs and their complexes].
The method of differential staining of muscles and connective tissue in the genitourinary organs (urinary bladder, ureters, renal pelvis) and gastrointestinal tract is suggested. The method consists of the intermittent use of known tissue van Gieson dyes (picric acid and fuchsin). When applied at pH = 7.2-8.0 (phosphate or citrate buffer) they stain the connective tissue in red and muscles in gold-yellow colour. A saturated solution of picric acid was mixed at equal proportions with water-free glycerol. Fuchsin was taken as 0.3% buffered solution or as 1% solution applied on the organ intermittently with picric acid. The organs, before staining, are fixed in 0.3 alcoholic solution of carbonic acid.


Effects of an Electronic Health Record Tool on Team Communication and Patient Mobility: A 2-Year Follow-up Study.
Intensive care unit early mobility programs improve patients' functional status and outcomes. An electronic health record-based communication tool improved interprofessional communication within an early mobility program. Long-term sustainability of this communication tool has not been evaluated. Ineffective interprofessional communication was perceived as a barrier to success of an intensive care unit early mobility program at the project institution. An electronic health record-based communication tool was successfully implemented to improve communication. Sustaining the intervention is of continued importance. Longitudinal data were collected 2 years after initial implementation of the communication tool to evaluate its continued impact on patient outcomes and staff engagement with an intensive care unit early mobility program. Initial implementation of the electronic health record-based communication tool resulted in statistically and clinically significant improvements in patient and staff metrics. Compared with postimplementation data, 2-year longitudinal follow-up data revealed nonsignificant changes in patient outcomes (time from admission to mobility goal, mechanical ventilation duration, length of intensive care unit stay, and discharge disposition recommendations to higher independence levels). Staff reported continued use of the communication tool and positive perceptions of its impact on the intensive care unit early mobility program. Nonsignificant changes in patient outcomes may indicate sustainment of the effect of the communication tool's original implementation. Employing appropriate sustainment techniques is essential to maintain practice change. The electronic health record-based communication tool can improve interprofessional communication within an intensive care unit early mobility program, improving patient outcomes and staff teamwork.


Development of a risk engine relating maternal glycemia and body mass index to pregnancy outcomes.
To develop a risk "engine" or calculator, integrating the risks of hyperglycemia, maternal BMI and other basic demographic data commonly available at the time of the pregnancy oral glucose tolerance test (OGTT), to predict an individual's absolute risk of specific adverse pregnancy outcomes. Data from the Brisbane HAPO cohort was analysed using logistic regression to determine the relationship between four clinical outcomes (primary CS, birth injury, large-for-gestational age, excess neonatal adiposity) with different combinations of OGTT results and maternal demographics (age, height, BMI, parity). Existing sets of international GDM diagnostic criteria were also applied to the cohort. 191 (15.3%) women were diagnosed as GDM by one or more existing criteria. All international criteria performed poorly compared to risk models utilising OGTT results only, or OGTT results in combination with maternal demographics. The risk engine's empirical performance on receiver - operator curve analysis was superior to the existing GDM diagnostic criteria tested. This concept shows promise for use in clinical practice, but further development is required.


Effect of retinoic acid and vitamin D on the expression of interleukin-1 beta, tumour necrosis factor-alpha and interleukin-6 in the human monocytic cell line U937.
We have previously described a synergism between the two physiological hormones, retinoic acid (RA) and 1 alpha,25-dihydroxyvitamin D3 (VD) in the induction of U937 cell differentiation towards a more mature state. Herein, we investigated the regulation of cytokine production during RA and/or VD treatment of U937 cells. Cell differentiation was followed by measurement of their capacity to give oxidative responses, and interleukin-1 beta (IL-1 beta), tumour necrosis factor-alpha (TNF-alpha) and IL-6 gene and protein expression were determined in RA/VD-treated cells, activated or not with lipopolysaccharide (LPS). The undifferentiated and RA-treated U937 cells were unable to produce monokines even when they were stimulated by LPS. VD induced the monokine mRNA expression in U937 cells but failed to induce protein release. However, unlike RA, it primed the cells to secrete monokines upon endotoxin stimulation. A large enhancement of the production of the monokines both at mRNA and protein levels was observed in the U937 cells exposed to the combination of RA + VD. Nevertheless, protein release required a further step of activation of the RA + VD-primed cells. The co-inducer effect of RA and VD was not observed in HL-60 or THP-1 cells and seems to be restricted to U937 cells. These results on cytokine expression support our previous finding that a combination of RA and VD brings the U937 cells to a high stage of myeloid differentiation with major characteristics of monocytes/macrophages.


Chamomile tea improves glycemic indices and antioxidants status in patients with type 2 diabetes mellitus.
Oxidative stress is a major factor in the pathogenesis of diabetes complications. The objectives were to investigate the effects of chamomile tea consumption on glycemic control and antioxidant status in subjects with type 2 diabetes mellitus (T2 DM). This single-blind randomized controlled clinical trial was conducted on 64 subjects with T2 DM (males and females) ages 30 to 60 y. The intervention group (n = 32) consumed chamomile tea (3 g/150 mL hot water) 3 times per day immediately after meals for 8 wk. The control group (n = 32) followed a water regimen for same intervention period. Fasting blood samples, anthropometric measurements, and 3-d, 24-h dietary recalls were collected at the baseline and at the end of the trial. Data were analyzed by independent t test, paired t test, and analysis of covariance. Chamomile tea significantly decreased concentration of glycosylated hemoglobin, serum insulin levels, homeostatic model assessment for insulin resistance, and serum malondialdehyde, compared with control group (all P < 0.05). Total antioxidant capacity, superoxide dismutase, glutathione peroxidase, and catalase activities were significantly increased by 6.81%, 26.16 %, 36.71 % and 45.06% respectively in chamomile group compared with these variables in control group at the end of the intervention (all P < 0.05). Short term intake of chamomile tea has beneficial effects on glycemic control and antioxidant status in patients with T2 DM. A larger sample population and a longer intervention period may be required to show significant clinical improvements.


Endothelial cells and hyperthermia.
Most radiation oncologists are aware of the effects of clinical hyperthermia on neoplastic cells. Its effects on blood vessels, however, are not as well recognized. Yet, since the 1960s a number of investigators have described and categorized the effects of hyperthermia on microvessels (in vivo), and on cultured endothelial cells (EC) (in vitro). Both EC and microvessels can be lethally damaged by the hyperthermia doses used as antineoplastic therapy. In vitro data indicate that capillary EC are moderately sensitive to hyperthermia. Proliferating EC are more thermosensitive suggesting that microvessels of malignant neoplasms (which contain many proliferating EC) are more affected than microvessels of normal tissues. This differential sensitivity of microvessels has also been observed in blood flow studies. Furthermore, hyperthermia inhibits angiogenesis. Thus, some of the antineoplastic effects of heat are caused by ischaemia due to obstruction or destruction of the tumour vessels or to inability to form new vessels. Sublethal EC damage can also be demonstrated, resulting in decreased synthesis of most proteins including adhesion molecules (as well as increased expression of a few such as heat shock proteins) and producing reversible loss of cytoskeletal elements. The therapeutic advantage provided by the higher thermal sensitivity of neoplastic vessels should be exploited further, perhaps by developing strategies specifically aimed to the tumour microvasculature.


Reduction of immunocompetent T cells followed by prolonged lymphopenia in severe sepsis in the elderly.
To investigate the immunological changes caused by severe sepsis in elderly patients. One-year, prospective observational study. Emergency department and intensive care unit of a single university hospital. Seventy-three patients with severe sepsis and 72 healthy donors. In elderly septic patients (aged 65 yr and over), 3-month survival was significantly reduced compared with that for adult patients (18-64 yr) (60% vs. 89%, p < 0.01). We found that lymphopenia was prolonged for at least 21 days in elderly nonsurvivors of sepsis, while the number of lymphocytes recovered in both adult and elderly survivors of sepsis. In order to examine the immunological status of septic patients, blood samples were collected within 48 hrs of diagnosis of severe sepsis, and peripheral blood mononuclear cells were purified for flow cytometric analysis. T cell levels were significantly reduced in both adult and elderly septic patients, compared with those in healthy donors (56% and 57% reduction, respectively). Interestingly, the immunocompetent CD28+ subset of CD4+ T cells decreased, whereas the immunosuppressive PD-1+ T cells and the percentage of regulatory T cells (CD4+ T cells that are both Foxp3+ and CD25+) increased in elderly patients, especially nonsurvivors, presumably reflecting the initial signs of immunosuppression. Reduction of immunocompetent T cells followed by prolonged lymphopenia may be associated with poor prognosis in elderly septic patients.


Ethnic bias amongst medical students in Aotearoa/New Zealand: Findings from the Bias and Decision Making in Medicine (BDMM) study.
Although health provider racial/ethnic bias has the potential to influence health outcomes and inequities, research within health education and training contexts remains limited. This paper reports findings from an anonymous web-based study examining racial/ethnic bias amongst final year medical students in Aotearoa/New Zealand. Data from 302 students (34% of all eligible final year medical students) were collected in two waves in 2014 and 2015 as part of the Bias and Decision Making in Medicine (BDMM) study. Two chronic disease vignettes, two implicit bias measures, and measures of explicit bias were used to assess racial/ethnic bias towards New Zealand European and Māori (indigenous) peoples. Medical students demonstrated implicit pro-New Zealand European racial/ethnic bias on average, and bias towards viewing New Zealand European patients as more compliant relative to Māori. Explicit pro-New Zealand European racial/ethnic bias was less evident, but apparent for measures of ethnic preference, relative warmth, and beliefs about the compliance and competence of Māori patients relative to New Zealand European patients. In addition, racial/ethnic bias appeared to be associated with some measures of medical student beliefs about individual patients by ethnicity when responding to a mental health vignette. Patterning of racial/ethnic bias by student characteristics was not consistent, with the exception of some associations between student ethnicity, socioeconomic background, and racial/ethnic bias. This is the first study of its kind with a health professional population in Aotearoa/New Zealand, representing an important contribution to further understanding and addressing current health inequities between Māori and New Zealand European populations.


Array comparative genomic hybridization analyses of all blastomeres of a cohort of embryos from young IVF patients revealed significant contribution of mitotic errors to embryo mosaicism at the cleavage stage.
Embryos produced by in vitro fertilization (IVF) have a high level of aneuploidy, which is believed to be a major factor affecting the success of human assisted reproduction treatment. The aneuploidy rate of cleavage stage embryos based on 1-2 biopsied blastomeres has been well-reported, however, the true aneuploidy rate of whole embryos remain unclear because of embryo mosaicism. To study the prevalence of mosaicism in top quality IVF embryos, surplus embryos donated from young patients (aged 28-32) in the assisted reproduction program at Queen Mary Hospital, Hong Kong were used. Thirty-six good quality day 2 embryos were thawed. Out of the 135 blastomeres in these embryos, 121 (89.6%) survived thawing. Twelve of these embryos without lysed blastomeres and which cleaved to at least seven cells after a 24-h culture were dissembled into individual blastomeres, which were analysed by array comparative genomic hybridization and microsatellite marker analysis by fluorescent PCR. Out of 12 day-3 embryos, 2 (16.7%) were normal, 3 (25%) were diploid/aneuploidy with <38% abnormality, 4 (33.3%) were diploid/aneuploidy mosaic with > =38% abnormality, and three (25%) were mosaic aneuploids. Conclusive chromosomal data were obtained from a high percentage of blastomeres (92.8%, 90/97). Microsatellite marker analysis performed on blastomeres in aneuploid embryos enabled us to reconstruct the chromosomal status of the blastomeres in each cleavage division. The results showed the occurrence of meiotic errors in 3 (25%) of the studied embryos. There were 16 mitotic errors (18.8%, 16/85) in the 85 mitotic divisions undertaken by the studied embryos. The observed mitotic errors were mainly contributed by endoreduplication (31.3%, 5/16), non-disjunction (25%, 4/16) and anaphase lagging (25%, 4/16). Chromosome breakages occurred in 6 divisions (7.1%, 6/85). Mosaicism occurs in a high percentage of good-quality cleavage stage embryos and mitotic errors contribute significantly to the abnormality.


From Ylides to Doubly Yldiide-Bridged Iron(II) High Spin Dimers via Self-Protolysis.
A synthetic strategy for the preparation of novel doubly yldiide bridged iron(II) high spin dimers ([(μ2-C)FeL]2, L = N(SiMe3)2, Mesityl) has been developed. This includes the synthesis of ylide-iron(II) monomers [(Ylide)FeL2] via adduct formation. Subsequent self-protolysis at elevated temperatures by in situ deprotonation of the ylide ligands results in a dimerization reaction forming the desired bridging μ2-C yldiide ligands in [(μ2-C)FeL]2. The comprehensive structural and electronic analysis of dimers [(μ2-C)FeL]2, including NMR, Mössbauer, and X-ray spectroscopy, as well as X-ray crystallography, SQUID, and DFT calculations, confirm their high-spin FeII configurations. Interestingly, the Fe2C2 cores display very acute Fe-C-Fe angles (averaged: 78.6(2)°) resulting in short Fe···Fe distances (averaged: 2.588(2) Å). A remarkably strong antiferromagnetic coupling between the Fe centers has been identified. Strongly polarized Fe-C bonds are observed where the negative charge is mostly centered at the μ2-C yldiide ligands.


[Biological risk factors for differentiated thyroid cancer].
Apart from the two relatively well documented risk factors for thyroid cancer i.e. iodine deficiency and ionising radiation, the mechanism of carcinogenesis involves a number of biological and environmental factors. The aim of the study was to analyse the selected biological and environmental risk factors in patients with differentiated thyroid cancer in comparison with a reference group in a case-control study. Both the group of 188 women and 30 men with thyroid cancer including 92 with follicular carcinoma and 126 with papillary carcinoma and the control group of 287 women and 58 men were subjected to the examination. All the study participants were given a standard questionnaire and underwent physical examination including thyroid USG, blood hormone levels and urine iodine concentration. The following risk factors were included in the analysis: family history, sex, age, thyroid morbidity, employment, body weight, hormone replacement therapy in women, cigarette smoking. In both groups with thyroid cancer women predominant with the female to male ratio 6.7 in follicular carcinoma and 6.0 in the papillary variant. In both groups of diagnoses the mean age was high--above 50, higher in men than in women. Over 70% of the patients with thyroid cancer had nodular goitre before the detection of cancer. In the control group the proportion of diagnosed goitre was slightly above 10%. A significantly more frequent hormone replacement therapy and contraception and more frequent miscarriages in women with thyroid cancer were noteworthy. One may assume that initiation, promotion and progression of the neoplastic process in the thyroid is triggered by a number of factors, both biological and environmental, and only multifactorial analysis may provide an insight into the mechanisms of carcinogenesis followed by initiation of preventives measures in appropriate cases.


A clinical study on insomnia in patients with cancer during chemotherapy containing high-dose glucocorticoids.
In this prospective, open-labeled study, 240 cancer patients were assigned to either a high-dose glucocorticoids (HDG) group that received chemotherapy containing HDG, or a control group that received chemotherapy without glucocorticoids. The Pittsburgh Sleep Quality Index (PSQI) was chosen to assess insomnia. The results of the study showed that dimensions of sleep latency, sleep duration, and sleep efficiency had the three largest differences in values and numbers of patients, with a score increase in the HDG group compared to the control group (p < 0.001). After chemotherapy in the HDG group, the PSQI score significantly increased in patients with stage II cancer (both p < 0.05), and patients diagnosed with lymphoma (p < 0.01), whereas the complete response and partial response rates (p < 0.05) had the smallest elevations. The average score of each dimension did not significantly decrease after hypnotics (p > 0.05). Our study suggests that the major clinical manifestations of insomnia in cancer patients receiving chemotherapy containing HDG include difficulty falling asleep, short sleep duration, and low sleep efficiency. however, we cannot definitively state that hypnotics can improve poor sleep quality.


Design, synthesis and evaluation of carbamate-linked uridyl-based inhibitors of human ST6Gal I.
Sialic acid at the terminus of cell surface glycoconjugates is a critical element in cell-cell recognition, receptor binding and immune responses. Sialyltransferases (ST), the enzymes responsible for the biosynthesis of sialylated glycans are highly upregulated in cancer and the resulting hypersialylation of the tumour cell surface correlates strongly with tumour growth, metastasis and drug resistance. Inhibitors of human STs, in particular human ST6Gal I, are thus expected to be valuable chemical tools for the discovery of novel anticancer drugs. Herein, we report on the computationally-guided design and development of uridine-based inhibitors that replace the charged phosphodiester linker of known ST inhibitors with a neutral carbamate to improve pharmacokinetic properties and synthetic accessibility. A series of 24 carbamate-linked uridyl-based compounds were synthesised by coupling aryl and hetaryl α-hydroxyphosphonates with a 5'-amino-5'-deoxyuridine fragment. The inhibitory activities of the newly synthesised compounds against recombinant human ST6Gal I were determined using a luminescent microplate assay, and five promising inhibitors with Ki's ranging from 1 to 20 µM were identified. These results show that carbamate-linked uridyl-based compounds are a potential new class of readily accessible, non-cytotoxic ST inhibitors to be further explored.


Construction of a synthetic Holliday junction analog and characterization of its interaction with a Saccharomyces cerevisiae endonuclease that cleaves Holliday junctions.
We describe the construction and characterization of an oligonucleotide Holliday junction analog and characterize its interaction with a Saccharomyces cerevisiae endonuclease that cleaves Holliday junctions. A Holliday junction analog containing four duplex arms and 54 base pairs was constructed by annealing four unique synthetic oligonucleotides. Mixing curve analysis showed that the complex contained a 1:1:1:1 mol ratio of the four unique sequence strands. In addition, a linear duplex with a sequence identical to two of the junction arms was also constructed for use as a control fragment. High resolution gel exclusion chromatography was used to purify and characterize the synthetic junction. The synthetic Holliday junction was found to be a specific inhibitor of a S. cerevisiae enzyme that catalyzes the cleavage of Holliday junctions. Under standard cleavage conditions, 50% inhibition was observed at a synthetic Holliday junction to substrate ratio of 7/1, whereas no inhibition by linear duplex was observed at molar ratios in excess of 150/1. Kinetic analysis showed that Holliday junction was a competitive inhibitor of the reaction and had an apparent Ki = 2.5 nM, although the mode of inhibition was complex. The synthetic Holliday junction was not a substrate for the enzyme, but was found to form a specific complex with the enzyme as evidenced by polyacrylamide gel electrophoresis DNA binding assays.


A Bright Future for Serial Femtosecond Crystallography with XFELs.
X-ray free electron lasers (XFELs) have the potential to revolutionize macromolecular structural biology due to the unique combination of spatial coherence, extreme peak brilliance, and short duration of X-ray pulses. A recently emerged serial femtosecond (fs) crystallography (SFX) approach using XFEL radiation overcomes some of the biggest hurdles of traditional crystallography related to radiation damage through the diffraction-before-destruction principle. Intense fs XFEL pulses enable high-resolution room-temperature structure determination of difficult-to-crystallize biological macromolecules, while simultaneously opening a new era of time-resolved structural studies. Here, we review the latest developments in instrumentation, sample delivery, data analysis, crystallization methods, and applications of SFX to important biological questions, and conclude with brief insights into the bright future of structural biology using XFELs.


Binder-free V2O5 cathode for greener rechargeable aluminum battery.
This letter reports on the investigation of a binder-free cathode material to be used in rechargeable aluminum batteries. This cathode is synthesized by directly depositing V2O5 on a Ni foam current collector. Rechargeable aluminum coin cells fabricated using the as-synthesized binder-free cathode delivered an initial discharge capacity of 239 mAh/g, which is much higher than that of batteries fabricated using a cathode composed of V2O5 nanowires and binder. An obvious discharge voltage plateau appeared at 0.6 V in the discharge curves of the Ni-V2O5 cathode, which is slightly higher than that of the V2O5 nanowire cathodes with common binders. This improvement is attributed to reduced electrochemical polarization.


Lactic acid fermentation stimulated iron absorption by Caco-2 cells is associated with increased soluble iron content in carrot juice.
An in vitro digestion/Caco-2 cell model was applied to explore the impact of lactic acid (LA) fermentation by Lactobacillus pentosus FSC1 and Leuconostoc mesenteroides FSC2 on the Fe bioavailability of carrot juice. The redox state of Fe in fermented carrot juice was also assessed as a crucial factor for absorption. LA fermentation was shown to improve mineral solubility to different extents at simulated physiological conditions: Mn (2-fold); Fe (1.5-1.7-fold); Zn (1.2-fold); Cu (1-fold). Soluble Fe2+ was increased about 16-fold by LA fermentation, and about one third of the Fe2+ remained soluble after in vitro digestion (about 4-5-fold higher than in fresh juice). Data on cell-line studies showed a 4-fold increase in the efficiency of Fe uptake, but not in transepithelial transfer by Caco2 cells, as a result of fermentation. The increases in Fe2+ level and the efficiency of cellular Fe uptake were strain-dependent. To sum up the effect on both Fe solubility and cellular uptake efficiency, the amount of cellularly absorbed Fe from Ln. mesenteroides FSC2-fermented juice was about 20 % higher than that from L. pentosus FSC1-fermented juice (22.7 v. 19.2 microg/l juice per mg protein). To conclude, LA fermentation enhanced Fe absorption by Caco-2 cells from carrot juice because of increases in not only Fe solubility after digestion, but also the efficiency of cellular Fe uptake. The fermentation-improved efficiency of Fe uptake was possibly due to the increased level of soluble Fe2+ rather than a being a strain-specific event.


Mutants generated by the insertion of random oligonucleotides into the active site of the beta-lactamase gene.
We have remodeled the gene coding for beta-lactamase by replacing DNA at the active site with random nucleotide sequences. The oligonucleotide replacement (Phe66XXXSer70XXLys73) preserves the codon for the active serine-70 but also contains 15 base pairs of chemically synthesized random sequences that code for 2.5 x 10(6) amino acid substitutions. From a population of Escherichia coli infected with plasmids containing these random inserts, we have selected seven new active-site mutants that render E. coli resistant to carbenicillin and a series of related analogues. Each of the new mutants contains multiple nucleotide substitutions that code for different amino acids surrounding serine-70. Each of the mutants exhibits a temperature-sensitive beta-lactamase activity. This technique offers the possibility of constructing alternative active sites in enzymes on the basis of biological selection for functional variants.


Evaluation of response rates to various antitumor agents of human gastric tumors implanted in nude mouse.
To ascertain the clinical predictability and, in the long run, the usefulness of the human tumor/nude mouse model as a screening tool for antitumor agents, it seems particularly important to use as many tumor lines as possible and to evaluate the therapeutic effectiveness of antitumor agents in terms of the overall response rate of a range of tumors. In this study, using 11 strains of established human gastric tumor xenografts with various histological characteristics and proliferation behavior, the experimental response rates to 8 typical antitumor drugs (mitomycin C, cyclophosphamide, ACNU, cisplatin, adriamycin, vincristine, vinblastine and 5-fluorouracil) were determined and compared with the clinical values. The experimental response rates to adriamycin, vincristine and 5-fluorouracil were in good accordance with the clinical results. However, with the other drugs, significantly higher response rates were observed with the nude mouse model as compared to clinical therapy, indicating that this model tends to overestimate the responsiveness of human tumor to a number of antitumor agents. These results strongly suggest the importance of using appropriate dose levels in the nude mouse to reproduce clinically equivalent effects in this model.


[An autopsy case of carbon dioxide intoxication].
A 44-year-old male, engaged in the transportation of dry ice, was found dead on the floor of the freezer of his refrigerator car which was parked in front of the place where he was scheduled to make a delivery. Autopsy was performed to investigate the cause of death. Apart from the signs of acute death, no other significant findings were obtained, either macro or microscopically; carbon dioxide poisoning was thus strongly suspected. We created a simulation experiment by using the refrigerator car to reproduce the events of the accident. The oxygen concentration in the freezer was 21.0% as indicated by oxygen sensors, but decreased to 17.1-17.4% when the engine was stopped. This decrease in oxygen concentration supposedly results from the production of carbon dioxide by the vaporization of dry ice. Carbon dioxide concentration in the air could be calculated from the change in the oxygen concentration in the closed space of the freezer. The concentration was assessed at 17.1-18.6%. An oxygen concentration of 17.1-17.4% does not of itself cause serious hypoxia, but a carbon dioxide concentration of 17.1-18.6% probably causes serious intoxication, because this value is beyond that of intoxication levels published in references. Therefore, we concluded that the cause of death in this case was carbon dioxide intoxication.


Infection trials in mice suggest that Macrorhabdus ornithogaster is not capable of growth in mammals.
The objective of this study was to determine if Macrorhabdus ornithogaster, a yeast recovered from the junction of the proventriculus and ventriculus of the stomach of parrots and other birds, can infect mice. Fifteen healthy ARC (S) female mice (age 10 weeks) were incoculated with M. ornithogaster by gavage at (two different dosage rates [n = 5]) or intraperitoneal injection (n = 5) (one dosage rate). They were euthanized 5 days later and examined for gross and microscopic evidence of infection. Macrorhabdus ornithogaster was not found to colonize the stomach, peritoneum, or viscera of the challeneged mice. The results of this study show that M. ornithogaster is not able to infect mice by the oral and intraperitoneal routes of administration and suggest that infection in mammals is unlikely to occur.


Renal failure in malaria.
Acute renal failure (ARF) is seen mostly in Plasmodium falciparum infection, but P vivax and P. malariae can occasionally contribute for renal impairment. Malarial ARF is commonly found in non-immune adults and older children with falciparum malaria. Occurance of ARF in severe falciparum malaria is quite common in southeast Asia and Indian subcontinent where intensity of malaria transmission is usually low with occasional microfoci of intense transmission. Since precise mechanism of malarial ARF is not known, several hypotheses including mechanical obstruction by infected erythrocytes, immune mediated glomerular and tubular pathology, fluid loss due to multiple mechanisms and alterations in the renal microcirculation, etc, have been proposed. Increased fluid administration, oxygen toxicity, and yet unidentified factors may contribute to pulmonary edema, acute respiratory distress syndrome (ARDS), multiorgan failure and death. Mainstay of treatment consists of appropriate antimalarial drug therapy, fluid replacement, and renal replacement therapy. Loop diuretics can convert an oliguric renal failure to non-oliguric renal failure without affecting outcome of the disease though the conversion reduces the risk of volume overload. There is little evidence on beneficial effect of vasoactive drugs. Nephrotoxic drugs such as ACE inhibitors, NSAIDs, aminoglycosides, cephalosporins should be avoided. Currently, high quality intensive care, early institution of renal replacement therapy, and avoidance of nephrotoxic drugs are standard practice of the prevention and management of ARF.


NMR analysis of quinocycline antibiotics: structure determination of kosinostatin, an antitumor substance from Micromonospora sp. TP-A0468.
A quinocycline antibiotic, kosinostatin, was isolated from the culture broth of Micromonospora sp. TP-A0468 along with isoquinocycline B. Structure of kosinostatin was determined to be the stereoisomer of isoquinocycline B regarding to the stereochemistry at the C-2' spiro carbon by NMR analysis. Kosinostatin isomerizes to isoquinocycline B through the inversion of the stereocenter at C-2'. Comparison of physico-chemical properties indicated that kosinostatin is presumably identical with quinocycline B isolated by CELMER et al. from Streptomyces aureofaciens.


A PDDA/poly(2,6-pyridinedicarboxylic acid)-CNTs composite film DNA electrochemical sensor and its application for the detection of specific sequences related to PAT gene and NOS gene.
2,6-Pyridinedicarboxylic acid (PDC) was electropolymerized on the glassy carbon electrode (GCE) surface combined with carboxylic group-functionalized single-walled carbon nanotubes (SWNTs) by cyclic voltammetry (CV) to form PDC-SWNTs composite film, which was rich in negatively charged carboxylic group. Then, poly(diallyldimethyl ammonium chloride) (PDDA), a linear cationic polyelectrolyte, was electrostatically adsorbed on the PDC-SWNTs/GCE surface. DNA probes with negatively charged phosphate group at the 5' end were immobilized on the PDDA/PDC-SWNTs/GCE due to the strong electrostatic attraction between PDDA and phosphate group of DNA. It has been found that modification of the electrode with PDC-SWNTs film has enhanced the effective electrode surface area and electron-transfer ability, in addition to providing negatively charged groups for the electrostatic assembly of cationic polyelectrolyte. PDDA plays a key role in the attachment of DNA probes to the PDC-SWNTs composite film and acts as a bridge to connect DNA with PDC-SWNTs film. The cathodic peak current of methylene blue (MB), an electroactive label, decreased obviously after the hybridization of DNA probe (ssDNA) with the complementary DNA (cDNA). This peak current change was used to monitor the recognition of the specific sequences related to PAT gene in the transgenic corn and the polymerase chain reaction (PCR) amplification of NOS gene from the sample of transgenic soybean with satisfactory results. Under optimal conditions, the dynamic detection range of the sensor to PAT gene target sequence was from 1.0x10(-11) to 1.0x10(-6) mol/L with the detection limit of 2.6x10(-12) mol/L.


[A frequent problem in the laboratory control of heparinization: contamination of blood specimens with exogenous heparin].
If blood for monitoring of heparin therapy is collected through indwelling catheters it may be contaminated by exogenous heparin. A prospective study comparing thrombin times in plasma obtained by venipuncture and by collection through heparin perfused catheters showed that 26 out of 77 catheter samples were contaminated. In 16 of these 26 cases, overestimation of the heparin effect would have led to incorrect dosage recommendations. It is concluded that blood for laboratory monitoring of heparin treatment should be collected by venipuncture.


Endotheliopathy in COVID-19-associated coagulopathy: evidence from a single-centre, cross-sectional study.
An important feature of severe acute respiratory syndrome coronavirus 2 pathogenesis is COVID-19-associated coagulopathy, characterised by increased thrombotic and microvascular complications. Previous studies have suggested a role for endothelial cell injury in COVID-19-associated coagulopathy. To determine whether endotheliopathy is involved in COVID-19-associated coagulopathy pathogenesis, we assessed markers of endothelial cell and platelet activation in critically and non-critically ill patients admitted to the hospital with COVID-19. In this single-centre cross-sectional study, hospitalised adult (≥18 years) patients with laboratory-confirmed COVID-19 were identified in the medical intensive care unit (ICU) or a specialised non-ICU COVID-19 floor in our hospital. Asymptomatic, non-hospitalised controls were recruited as a comparator group for biomarkers that did not have a reference range. We assessed markers of endothelial cell and platelet activation, including von Willebrand Factor (VWF) antigen, soluble thrombomodulin, soluble P-selectin, and soluble CD40 ligand, as well as coagulation factors, endogenous anticoagulants, and fibrinolytic enzymes. We compared the level of each marker in ICU patients, non-ICU patients, and controls, where applicable. We assessed correlations between these laboratory results with clinical outcomes, including hospital discharge and mortality. Kaplan-Meier analysis was used to further explore the association between biochemical markers and survival. 68 patients with COVID-19 were included in the study from April 13 to April 24, 2020, including 48 ICU and 20 non-ICU patients, as well as 13 non-hospitalised, asymptomatic controls. Markers of endothelial cell and platelet activation were significantly elevated in ICU patients compared with non-ICU patients, including VWF antigen (mean 565% [SD 199] in ICU patients vs 278% [133] in non-ICU patients; p<0·0001) and soluble P-selectin (15·9 ng/mL [4·8] vs 11·2 ng/mL [3·1]; p=0·0014). VWF antigen concentrations were also elevated above the normal range in 16 (80%) of 20 non-ICU patients. We found mortality to be significantly correlated with VWF antigen (r = 0·38; p=0·0022) and soluble thrombomodulin (r = 0·38; p=0·0078) among all patients. In all patients, soluble thrombomodulin concentrations greater than 3·26 ng/mL were associated with lower rates of hospital discharge (22 [88%] of 25 patients with low concentrations vs 13 [52%] of 25 patients with high concentrations; p=0·0050) and lower likelihood of survival on Kaplan-Meier analysis (hazard ratio 5·9, 95% CI 1·9-18·4; p=0·0087). Our findings show that endotheliopathy is present in COVID-19 and is likely to be associated with critical illness and death. Early identification of endotheliopathy and strategies to mitigate its progression might improve outcomes in COVID-19. This work was supported by a gift donation from Jack Levin to the Benign Hematology programme at Yale, and the National Institutes of Health.


Geriatric Psychiatry Fellowship Recruitment: Crisis or Opportunity?
The increasing number of elderly patients with psychiatric disorders has led to a shortage of psychiatrists with specialty training in geriatric psychiatry. Despite the emergence of a critical mass of faculty and a steady growth in the number of fellowship training programs, increasing subspecialization in general psychiatry, a dwindling fellowship applicant pool, ageism, and financial concerns have contributed to a potential recruitment crisis. The author discusses such problems and describes strategies to solve them, including providing role models, developing award programs and affinity groups, securing program funding, sharing resources, refining program focus, balancing practice patterns, assisting in practice transitions, and marketing aggressively.


Phalloidin-induced cholestasis: a microfilament-mediated change in junctional complex permeability.
Phalloidin, administered to male rats for 7 days (500 microgram per kg/day), increased the mean hepatic content of filamentous actin. Both bile flow and bile acid excretion diminished proportionally, whereas the bile-to-plasma ratios of [3H]inulin and [14C]sucrose increased significantly from 0.08 and 0.16 in controls to 0.37 and 0.69, respectively, in phalloidin-treated animals. Simultaneously, junctional permeability was altered as noted by the free penetration of ionic lanthanum into the zonula occludens and bile canaliculus. Freeze-fracture replicas of the junctional complex revealed rearrangements of the junctional elements and regions in which only a single element separated the canaliculus from the lateral intercellular space. These findings suggest that microfilaments influence the permeability of "tight junctions" between hepatocytes and that bile constituents might reflux from the canaliculus to the intercellular space in phalloidin-induced cholestasis.


Epidemiological patterns and incidence of bio-, sero- and phage types of Vibrio cholerae in Hyderabad, India, during 1971-1984.
During 1971-1984 out of 44,762 gastroenteritis cases bacteriologically examined, 4240 (9.5%) Vibrio cholerae were isolated in Hyderabad, India. Out of them 1329 (31.3%) were classical and 2911 (68.7%) were el-tor biotypes. The changeover from classical to el-tor cholera was observed in Hyderabad during 1975 and persisted. During this 14 year period four major outbreaks with V. cholerae serogroup O1 Ogawa, V. cholerae biotype el-tor serogroup O1 Ogawa were observed. Phage types 1, 2 and 4 were prevalent during the classical period and types 2 and 4 during the el-tor period. Of the cholera patients, 67% were under 30 years of age with no significant difference in the incidence among females (51.4%) and males (48.6%). The outbreaks were of protracted pattern with only few cases per day or week with peak incidence during monsoon (May-August). Community of the low socio-economic strata was the most susceptible group. Five zones situated at the outlet of the Moosi river in the downhill were recorded as extensive cholera transmission zones and two of them were the primary foci of infection. The severity of the infection was found directly related to the average rainfall during the year leading to the sewage stagnation in the downhill.


Identification and characterization of a putative serine protease expressed in vivo by Mycobacterium avium subsp. paratuberculosis.
A putative serine protease expressed in vivo by Mycobacterium avium subsp. paratuberculosis was isolated from a lambda gt11 genomic expression library by screening with serum from a naturally infected sheep. The gene was contained in two overlapping clones, which were shown by antibody elution to encode a protein of 34 kDa in M. a. paratuberculosis. The clones were sequenced and database searches detected a motif identical to the active serine site in trypsin, and 30% homology to the putative serine proteases (HtrA proteins) of Escherichia coli, Salmonella typhimurium, Brucella abortus and Rochalimaea henselae.


[Speciation and characterization of interaction of PACls with sulfate].
The interaction of sulfate with various PACl was investigated by using Ferron assay, chemical analysis and SEM. The experimental results showed that the basicity (B = [OH]/[Al]) exhibited significant role in the PACl-sulfate reaction. It indicated different species in various PACl under different reaction pathway with sulfate. The Alc formed precipitation quickly with sulfate, while Alb underwent slowly crystalization. The decrease of Ala resulted in the limit of ferron method. The different speciation component in PACls formed different crystal morphology and chemical composition with sulfate. Increase the basicity, the content of sulfate in precipitate decreased from 0.45 to 0.30.


Host-Guest Interactions Between Metal-Organic Frameworks and Air-Sensitive Complexes at High Temperature.
The host-guest chemistry of metal-organic frameworks (MOFs) has been attracting increasing attention owing to the outstanding properties derived from MOFs-guests combinations. However, there are large difficulties involved in the syntheses of the host-guest MOF systems with air-sensitive metal complexes. In addition, the behaviors on host-guest interactions in the above systems at high temperature are not clear. This study reported the synthetic methods for host-guest systems of metal-organic framework and air-sensitive metal complexes via a developed chemical vapor infiltration process. With the synchrotron X-ray powder diffraction (XRPD) measurements and Fourier Transform infrared spectroscopy (FTIR), the successful loadings of Fe(CO)5 in HKUST-1 and NH2-MIL-101(Al) have been confirmed. At high temperatures, the structural and chemical componential changes were investigated in detail by XRPD and FTIR measurements. HKUST-1 was proven to have strong interaction with Fe(CO)5 and resulted in a heavy loading amount of 63.1 wt%, but too strong an interaction led to deformation of HKUST-1 sub-unit under heating conditions. NH2-MIL-101(Al), meanwhile, has a weaker interaction and is chemically inert to Fe(CO)5 at high temperatures.


D-TAG: erasing the tag of gang membership.
Gangs are noted for establishing their territory, flaunting gang affiliation, intimidating nonmembers, and documenting their "services performed." These examples are a few reasons for the practice of "tagging," the labeling of an area, person, or object with gang-related graffiti or markings, such as tattoos. This article describes a school nurse's response to gang "tagging" and her efforts to assist former gang members who request removal of their tattoos, to get them removed-in essence to D-TAG themselves from their gang affiliation. D-TAG is a volunteer rehabilitation program utilizing family and community interaction to support gang tattoo removal and direct activities away from gang affiliations toward alternative educational programs and life styles.


[The preventive and treatment tours as a part of psychological aid system provided for soldiers who participated in the military missions abroad].
The therapeutic and preventive tours were brought into being in 2004 due to the extension of the commitments made by Polish forces to conduct military activities abroad. They came into existence as an element of psychological aid system designed for soldiers who were deployed to missions outside Poland. The tours are conducted by psychologists in four military rehabilitation and treatment hospitals, according to the assumptions of the Antistress Programme. The participants of the tours are soldiers who returned from their deployments and who reveal precursory symptoms of stress disorders. Over 2500 participants have already taken part in such tours up till now, and every year the number of people referred to such tours increases. To gauge the participants' as well personnel's opinion about the efficiency of the therapeutic and preventive tours. 103 participants of tours which took place in August and September 2007 have been questioned. The examined mainly consist of men, aged from 25 till 45. The research method has been based on an anonymous questionnaire constructed by the author of this project. There have been formulated only hypotheses which can constitute the basis for further, more detailed studies. Over half of the questioned obtained information about the tours from a source different from doctors or psychologists. klost often the source of information was the Internet or colleagues who had already participated in such tours. The above mentioned sources have also determined the choice of a particular sanatorium. Apart from the obligatory preliminary consultations by the doctor and psychologist, almost all the participants of the tours participated in rehabilitation treatment. Less than half of the questioned attended relaxation sessions conducted by the psychologist. The soldiers tend to disregard or hide their symptoms of psychical nature, nevertheless their reaction to stress manifested in a form of somatic symptoms, has been accepted and so has the manner of the treatment applied. The majority of the questioned have estimated the whole of "Anti-stress Training" programme well or very well. A similar opinion has been formed with reference to the activities conducted during the tours by psychologists. The questioned have favourably assessed the function that the "Antistress Training" programme has had in the psychological support process. The system of information about the possibility to take advantage of the offer by the soldiers requires an improvement, though. What is more, the psychologists and the doctors should also be more involved in such activities. It is a significant task for the psychologists to inform soldiers about the range of psychological aid provided during the tours, which can result in better attendance at the activities as well as creating more positive approach to psychologists' work in general. The significant problem is how to convince the commanders of a higher rank of the effectiveness of programmes of such a type on soldiers'psychophysical condition.


Evaluating Family Medicine Resident Narrative Comments Using the RIME Scheme.
In 2013, the Accreditation Council on Graduate Medical Education (ACGME) launched the Next Accreditation System, which required explicit documentation of trainee competence in six domains. To document narrative comments, the University of North Carolina Family Medicine Residency Program developed a mobile application to document real time observations. The objective of this work was to assess if the Reporter, Interpreter, Manager, Expert (RIME) framework could be applied to the narrative comments in order to convey a degree of competency. From August to December 2020, 7 individuals analyzed narrative comments of four family medicine residents. The narrative comments were collected from July to December 2019. Each individual applied the RIME framework to the comments and the team met to discuss. Comments where 5/7 individuals agreed were not further discussed. All other comments were discussed until consensus was achieved. 102 unique comments were assessed. Of those comments, 25 (25.5%) met threshold for assessor agreement after independent review. Group discussion about discrepancies led to consensus about the appropriate classification for 92 (90.2%). General comments on performance were difficult to fit into the RIME framework. Application of the RIME framework to narrative comments may add insight into trainee progress. Further faculty development is needed to ensure comments have discrete elements needed to apply the RIME framework and contribute to overall evaluation of competence.


Current Status of and Perspectives on the Application of Marmosets in Neurobiology.
The common marmoset (Callithrix jacchus), a small New World primate, is receiving substantial attention in the neuroscience and biomedical science fields because its anatomical features, functional and behavioral characteristics, and reproductive features and its amenability to available genetic modification technologies make it an attractive experimental subject. In this review, I outline the progress of marmoset neuroscience research and summarize both the current status (opportunities and limitations) of and the future perspectives on the application of marmosets in neuroscience and disease modeling.


Pharmacokinetics of tulathromycin and its metabolite in swine administered with an intravenous bolus injection and a single gavage.
Tulathromycin is a macrolide antimicrobial agent proposed for therapeutic use in treatment of porcine and bovine respiratory disease. In this study, the absolute bioavailability of tulathromycin solution was investigated in pigs. Eight pigs, with body weight of 20.5 ± 1.6 kg, were given a single dose of tulathromycin at 2.5 mg/kg oral (p.o.) and intravenous (i.v.) in a crossover design. The plasma concentrations of tulathromycin and its metabolite were determined by LC-MS/MS method, and the pharmacokinetic parameters of tulathromycin were calculated by noncompartmental analysis. After p.o. administration, the maximum plasma concentration (C(max) ) was 0.20 ± 0.05 μg/mL at 3.75 ± 0.71 h. The terminal half-life (t(1/2λz) ) in plasma was 78.7 ± 6.75 h, and plasma clearance (Cl/F) was 1.14 ± 0.28 L/h/kg. After i.v. injection, plasma clearance (Cl) was 0.580 ± 0.170 L/h/kg, the volume of distribution (Vz) was 64.3 ± 21.2 L/kg, and the t(1/2λz) was 76.5 ± 13.4 h. In conclusion, an analytical method for the quantification of tulathromycin and its metabolite in plasma in swine was developed and validated. Following p.o. administration to pigs at 2.5 mg/kg b.w., tulathromycin was rapidly absorbed and the systemic bioavailability was 51.1 ± 10.2.


Enhanced Electrochemiluminescence of One-Dimensional Self-Assembled Porphyrin Hexagonal Nanoprisms.
In this work, we synthesized the one-dimensional nanostructure of zinc 5,10,15,20-tetra(4-pyridyl)-21H,23H-porphine (ZnTPyP) via a self-assembly technique. Using sodium dodecyl sulfate (SDS) as "soft template", the self-assembled ZnTPyP (SA-ZnTPyP) had the morphology of hexagonal nanoprisms with a uniform size (diameter of 100 nm). The SA-ZnTPyP exhibited remarkably different spectral properties compared to those of the original ZnTPyP. The as-prepared SA-ZnTPyP was used to modify glassy carbon electrodes (GCE), and the electrochemiluminescence (ECL) behaviors of the SA-ZnTPyP/GCE were investigated. The hydrophilic carbon dots (C-dots) could efficiently prevent the dissolution of SA-ZnTPyP in DMF containing 0.1 mol L-1 TBAP and, simultaneously, could accelerate electron transfer. Therefore, the enhanced ECL was realized by C-dots/SA-ZnTPyP/GCE by using H2O2 as co-reactant. This amplification of ECL was further studied by ECL spectroscopies and cyclic voltammetry, and the corresponding mechanism was proposed.


Chronic hypertension and vascular alterations following short-term high dietary salt intake in postweaning rats.
The chronic effects of short-term administration of a high salt diet to postweaning rats on blood pressure (BP) was studied. BP was significantly elevated following 4 weeks of treatment. After stopping the high salt diet, BP dropped to control levels but then progressively rose again to hypertensive levels. A significant increase in arterial medial thickness as well as an increase in contractile sensitivity and a decrease in relaxation responsiveness were observed in aortic smooth muscle. These data show that short-term administration of a high salt diet to normal postweaning rats results in a chronic elevation in BP accompanied by significant alterations in vascular structure and function.


Technologies of authority in the medical classroom in the thirteenth and fourteenth centuries.
By 1300, university medical masters were introducing their students to a culturally distinctive reality. This reality was based on the twin pillars sustaining institutional medical knowledge: authority and a logical apparatus based on Aristotelian principles. Traditionally, attention has been paid to the relationship of the medical author with his classical authorities. This paper analyzes the strategies developed by the university medical master for establishing himself as an authority, which entailed treating his contemporaries as authorities as well. It is suggested that a tendency can be traced in the medical classroom from the 1340s onwards to turn attention away from the classical authors towards contemporary writers.


Adamantinoma of long bones: a long-term follow-up study of 11 cases.
The aim of this study was to evaluate the clinicopathological features and prognostic significances of 11 histologically proven adamantinoma cases based on an average 12,7 year long follow-up. The male: female ratio was 8:3, aged between 4 and 80 years (mean 29,3 years). The initial diagnosis at referral was other than adamantinoma in six patients (fibrous dysplasia, carcinoma metastasis, osteofibrous dysplasia, bone cyst, non-ossifying fibroma), referring to the differential diagnostic problems. All tumors were localized to the mid part of tibia. By histological evaluation, basaloid pattern on a background of fibrotic stroma dominated in six patients, while spindle and squamous features were less frequently seen. All adamantinoma were positive for cytokeratins often in coexpression with vimentin. No correlation was experienced between histology and clinical outcome. Intralesional curettage (2 pts) was followed by recurrence of the tumor. Wide resection was performed in eight patients with reconstruction using intercalary fibula autografts in seven patients. Reconstruction-related complications occurred in two third of the cases, all of them could however be controlled by repeated surgery. Six recurrences occurred in four patients, two of these recurrences occurred 20 and 16 years after initial surgery. One patient died 9 years after recognition of the tumor of pulmonary metastases. Adamantinoma of the long bones is a low grade malignant tumor, which clinical outcome is difficult to predict based on histology or surgical stage of the tumor. Wide surgical margin, e.g. resection the tumor reduces the rate of recurrence. This study underlines that recurrences do occur even decades after recognition the tumor, therefore a life-long follow-up of the patient is necessary.


Prevalence of Positive Diabetes-Associated Autoantibodies among Type 2 Diabetes and Related Metabolic and Inflammatory Differences in a Sample of the Bulgarian Population.
The study aimed to estimate the prevalence of unrecognized cases with positive autoantibodies among type 2 diabetes (T2D) in a sample of the Bulgarian population and to compare some metabolic and inflammatory markers to those of patients having negative autoantibodies and subjects with latent autoimmune diabetes (LADA). Patients with T2D, patients with LADA, and control participants were enrolled. Antiglutamic acid decarboxylase, anti-insulinoma-associated 2, and antizinc transporter 8 autoantibodies were assayed through ELISA. C-reactive protein and interleukin 6 (IL-6) and tumor necrosis factor alpha were assessed. Ten percent of patients with T2D had positive autoantibodies. They had lower body mass index (p = 0.014), worse glycemic control (HbA1c, p = 0.033), and better HDL cholesterol (p = 0.026) than those in negative autoantibodies cases. Compared to LADA, glycemia and anthropometric data did not differ significantly but metabolic syndrome was more prevalent among newly found cases with positive autoantibodies (p = 0.046). Their level of inflammatory markers was similar to that of patients having negative autoantibodies (p > 0.05), but IL-6 was higher when compared to LADA (p = 0.002). Prevalence of patients having positive autoantibodies within T2D in the analyzed sample of the Bulgarian population was 10%. They shared common metabolic features with subjects with LADA, but inflammatory phenotype was closer to that of T2D.


Animal-assisted psychotherapy for young people with behavioural problems in residential care.
The aim of this study was to evaluate the impact of an animal-assisted psychotherapy (AAP) programme on clinical symptoms, personal adjustment, and adaptive skills in a group of adolescents in residential care who had experienced childhood trauma and who presented mental health problems and difficulties adapting to the care home environment. The 87 participants (Mage = 15.17, SD = 1.53) were divided into two groups: a treatment group (25 girls and 27 boys; Mage = 15.00, SD = 1.55) and a control group (9 girls and 26 boys; Mage = 15.42, SD = 1.50). The programme consisted of 34 sessions involving both group (23 sessions) and individual (11 sessions) AAP. The Behaviour Assessment System for Children was used to evaluate clinical and adaptive dimensions of behaviour and personality. The results indicated that, in comparison with controls, the young people who took part in the AAP programme reported a significant improvement on two measures of internalizing symptoms, namely, depression and sense of inadequacy. Although no significant differences were observed in relation to externalizing symptoms, the adolescents who received the AAP programme showed improved social skills in terms of their ability to interact satisfactorily with peers and adults in the care home environment, as well as a more positive attitude towards teachers at school. These results suggest that AAP may be a promising treatment for young people who have experienced childhood trauma and who subsequently find it difficult to adapt to the residential care setting.


Impact of right coronary dominance on triple-vessel coronary artery disease: A cross-sectional study.
This study was conducted to investigate the relationship between right coronary dominance and coronary angiographic characteristics in patients with or without significant coronary artery disease (CAD).A total of 2225 patients undergoing coronary angiography (CAG) between January 2011 and November 2014 were recruited in our study. Based on the CAG results, patients were divided into the left dominance (LD) group, right dominance (RD) group, and co-dominance (CD) group. Multinomial logistic regression was applied to analyze the relationships between coronary dominance and triple-vessel CAD.We found that patients with RD had a higher prevalence of triple-vessel CAD (36.6% vs 27.3%, P = .008) and significant stenosis in the right coronary artery (40.5% vs 29.2%, P = .001). In addition, results of multinomial logistic regression analysis showed that RD was significantly associated with the triple-vessel disease (odds ratio 1.768, 95% confidence interval 1.057-2.956, P = .030).In conclusion, RD positively correlated with triple-vessel CAD rather than LD or CD in patients. This result suggested that RD may serve as a risk factor for triple-vessel CAD and more effective measures should be taken in RD patients to prevent fatal cardiovascular events.


Caudal ropivacaine and bupivacaine for postoperative analgesia in infants undergoing lower abdominal surgery.
To compare the postoperative analgesic efficacy of ropivacaine 0.175% and bupivacaine 0.175% injected caudally into infants for lower abdominal surgery. Eighty infants, aged 3-12 months, ASA I-II scheduled to undergo lower abdominal surgery were randomly allocated to one of the two groups: Group R received 1ml.kg(-1) 0.175% ropivacaine and Group B received 1ml.kg(-1) 0.175% bupivacaine via caudal route. Postoperative analgesia, sedation and motor block were evaluated with modified objective pain scale, three-point scale and modified Bromage scale respectively. Postoperative measurements including mean arterial pressure (MAP), heart rate (HR), pain (OPS), sedation and motor block score were recorded for four hours in the postoperative recovery room. Parents were contacted by telephone after 24 hours to question duration of analgesia and side effects. No significant differences were found among the groups in demographic data, MAP, HR, OPS and sedation scores during four hours postoperatively. The duration of analgesia was 527.5±150.62 minutes in Group R, 692.77±139.01 minutes in Group B (p=0.004). Twelve (30%) patients in Group R, 16 (40%) patients in groupB needed rescue analgesics (p=0.348). Rescue analgesics were administered (1 time/2 times) (9/3) (22.5/7.5%) in Group R and 16/0 (40/0%) in Group B, where no statistically significant difference was determined between the groups (p=0.071). Motor blockade was observed in 7 (17.5%) patients in Group R, and 8 (20%) patients in Group B (p=0.774). This study indicated, that a concentration of 0.175% ropivacaine and 0.175% bupivacaine administered to the infants via caudal route both provided effective and similar postoperative pain relief in infants, who underwent lower abdominal surgery.


Inhibiting action of chlorophenols on biodegradation of phenol and its correlation with structural properties of inhibitors.
An activated sludge conditioned to a low concentration of phenol was used for the biodegradation of phenol in the presence of Chlorophenols, measuring the inhibiting action of all the latter compounds and expressing it as p/(50.) As a structural property to be correlated with the measured values, the lipophilicity (log P) of the Chlorophenols was reevaluated with a new apparatus. Values of p/(50)p are better correlated with the number of chloro substituents in the inhibitor, provided a specific effect of substituents ortho to the phenolic function is considered. New results are compared with previous measurements effected on a different activated sludge.


In Vitro Control of Uropathogenic Microorganisms with the Ethanolic Extract from the Leaves of Cochlospermum regium (Schrank) Pilger.
The roots of Cochlospermum regium, popularly known as "algodãozinho-do-cerrado," are used for the treatment of genitourinary infections. However, the removal of their subterranean structures results in the death of the plant, and the use of the leaves becomes a viable alternative. Therefore, the antimicrobial activity of Cochlospermum regium leaf's ethanolic extract and its action on the biofilm formation of microorganisms associated with urinary infection were evaluated. The total phenolic compounds, flavoids, and tannins were quantified using the reagents Folin-Ciocalteu, aluminum chloride, and vanillin, respectively. The antimicrobial activity was evaluated by the broth microdilution method and the effect of the extract in the biofilm treatment was measured by the drop plate method. Cytotoxicity was evaluated by the method based on the reduction of MTS and the mutagenicity by the Ames test. The ethanolic extract of C. regium leaves presented 87.4 mg/EQ of flavonoids, 167.2 mg/EAG of total phenolic compounds, and 21.7 mg/ECA of condensed tannins. It presented reduction of the biofilm formation for E. coli and C. tropicalis and antimicrobial action of 1 mg/mL and 0.5 mg/mL, respectively. The extract showed no cytotoxicity and mutagenicity at the concentrations tested. This study demonstrated that C. regium leaves are a viable option for the treatment of genitourinary infections and for the species preservation.


Photo-cross-linked poly(thioether-co-carbonate) networks derived from the natural product quinic acid.
Polycarbonate networks derived from the natural product quinic acid that can potentially return to their natural building blocks upon hydrolytic degradation are described herein. Solvent-free thiol-ene chemistry was utilized in the copolymerization of tris(alloc)quinic acid and a variety of multifunctional thiol monomers to obtain poly(thioether-co-carbonate) networks with a wide range of achievable thermomechanical properties including glass transition temperatures from -18 to +65 °C and rubbery moduli from 3.8 to 20 MPa. The network containing 1,2-ethanedithiol expressed an average toughness at 25 and 63 °C of 1.08 and 2.35 MJ/m(3), respectively, and an order-of-magnitude increase in the average toughness at 37 °C of 15.56 MJ/m(3).


Chemoproteomic Profiling of Cobalamin-Independent Methionine Synthases in Plants with a Covalent Probe.
Cobalamin-independent methionine synthases (MS) are zinc-binding methyltransferases that catalyze de novo methionine biosynthesis in higher plants, which are enzymes critically involved in seed germination and plant growth. Here, we report a highly selective sulfonyl fluoride-based probe for chemoproteomic profiling of MS enzymes in living systems of the model plant Arabidopsis thaliana, as implemented in in-gel-, mass spectrometry-, and imaging-based platforms. This probe holds promise for facilitating and accelerating fundamental research and industrial application of MS enzymes, particularly in the contexts of MS1/2-targeting herbicide screening and design.


Quantification of skeletal kinetic indices in Paget's disease using dynamic 18F-fluoride positron emission tomography.
The purpose of this study was to quantify indices of regional bone metabolism in Paget's disease and to compare these indices with normal bone using dynamic 18F-fluoride positron emission tomography (PET). Seven patients with vertebral Paget's disease had 1 h dynamic 18F-fluoride PET scans performed. The scans included a diseased vertebra and an adjacent normal vertebra. Arterial plasma input functions were also measured. A three-compartment, four-parameter model was used with nonlinear regression analysis to estimate bone kinetic variables. Compared with normal bone, pagetic bone demonstrated higher values of plasma clearance to bone mineral (Ki; 1.03 x 10(-1) vs. 0.36 x 10(-1) ml/min per milliliter; p = 0.018) and clearance to total bone tissue (K1; 2.38 x 10(-1) vs. 1.25 x 10(-1) ml/min per milliliter; p = 0.018), reflecting increased mineralization and blood flow, respectively. Release of 18F-fluoride from bone mineral (k4) was lower in pagetic bone (p = 0.022), suggesting tighter binding of 18F-fluoride to bone mineral. The notional volume of the extravascular bone compartment (K1/k2) was greater in pagetic bone (p = 0.018). Although the unidirectional extraction efficiency from the extravascular space to bone mineral (Ki/K1) was greater in pagetic bone (p = 0.018), a lower pagetic value of k2 (p = 0.028), describing the rate of transfer from the bone extravascular compartment to plasma, suggests that the 18F-fluoride that enters the relatively fibrotic marrow space of pagetic bone may be less accessible for return to plasma. These findings confirm some of the known pathophysiology of Paget's disease, introduce some new observations, and show how dynamic 18F-fluoride PET may be of value in the measurement of regional metabolic parameters in focal bone disorders.


Educating the nursing workforce in 2000 and beyond.
Changes in the health care delivery system have comparable impacts on both nursing practice and education, such that tomorrow's nursing practice would best emanate from innovative partnerships between leaders in practice and education. For the foreseeable future, an expert nursing workforce is needed in an expanded form, but an array of challenges to maintaining this workforce are evident. Recent and continuing scientific and technologic advances in health care make the matching of practice and education to evolving trends paramount. Regardless of what the future holds, generative leadership in any realm of practice is essential to move our profession into the forefront of health care.


Quantitative immunoelectron microscopic localization of (Na+,K+)ATPase in rat parotid gland.
Distribution of (Na+,K+)ATPase on the cell membranes of acinar and duct cells of rat parotid gland was investigated quantitatively by immunoelectron microscopy using the post-embedding protein A-gold technique. In acinar cells, ATPase was localized predominantly on the basolateral plasma membranes. A small but significant amount of (Na+,K+)ATPase was, however, detected on the luminal plasma membranes, especially on the microvillar region of the acinar cells; the surface density on the luminal membrane was approximately one third of that on the basolateral membranes. In duct cells, many gold particles were found on the basolateral membrane, especially along the basal infoldings of the plasma membranes, whereas no significant gold particles were found on the luminal plasma membranes, suggesting unilateral distribution of ATPase in duct cells. We suggest that in acinar cells sodium ion is not only transported paracellularly but is also actively transported intracellularly into the luminal space by the (Na+,K+)ATPase located on the luminal plasma membranes, and that water is passively transported to the luminal space to form a plasma-like isotonic primary saliva, while in the duct cells the same ion is selectively re-absorbed intracellularly by (Na+,K+)ATPase found in abundance along the many infoldings of the basal plasma membranes, thus producing the hypotonic saliva.


Abdominal tuberculosis after removal of an adjustable gastric band - report of an unusual case.
Laparoscopic adjustable gastric banding (LAGB) is the third most popular bariatric procedure. Despite its reversibility and minimal invasiveness, band infection affects 1.2% of patients. We present a case of a 25-year-old, obese woman who was experiencing malaise and feverishness 3 years after gastric band placement. Due to port site infection the port was removed, which did not improve the patient's condition. After 2 years the band was removed via laparotomy with a minor surgical site infection reported. The patient returned 2 weeks after discharge with signs of sepsis. After ruling out pulmonary causes, an exploratory laparotomy was performed, revealing granulomatous peritonitis. Standard histopathological examinations, broncho-alveolar lavage culture and DNA tests along with microbiological cultures were inconclusive. Broad-spectrum antibiotics and antifungal and antiparasitic agents did not improve the patient's condition. Mycobacterium tuberculosis DNA was discovered in a greater omentum specimen. The patient was treated with isoniazid, rifampicin, pyrazinamide and streptomycin for four months.


Venous ulceration.
In the last few years, interest in the pathogenesis and treatment of venous ulceration has been rekindled by novel observations in fibrinolysis and an explosive growth in wound care products, respectively. The purpose of this work is to summarize the salient clinical features of venous ulceration, its proposed pathogenesis, and conventional as well as future treatment modalities. Venous ulceration is an increasingly common condition. Its pathogenesis is related to failure of the calf muscle pump, and treatment is directed at effective leg compression, avoidance of contact dermatitis, and proper wound care. There is increased interest in both the pathogenic steps involved in venous ulceration and in treatment modalities. We suspect that incremental progress will occur in these areas in the next few years.


Comparison between local ablative therapy and chemotherapy for non-resectable colorectal liver metastases: a prospective study.
There is a growing interest for the use of local ablative techniques in patients with non-resectable colorectal liver metastases. Evidence on the efficacy over systemic chemotherapy is, however, extremely weak. In this prospective study we aim to assess the additional benefits of local tumour ablation. A consecutive series of 201 colorectal cancer patients, without extrahepatic disease, that underwent laparotomy for surgical treatment of liver metastases, were prospectively followed for survival and HRQoL. At laparotomy three groups were identified: patients in whom radical resection of metastases proved feasible, patients in whom resection was not feasible and received local ablative therapy, and patients in whom resection or local ablation was not feasible for technical reasons and who received systemic chemotherapy. Patients in the chemotherapy and in local ablation group were comparable for all prognostic variables tested. For the local ablation group overall survival at 2 and 5 years was 56 and 27%, respectively (median 31 months, n = 45), for the chemotherapy group 51 and 15%, respectively (median 26 months, n = 39) (P = 0.252). After resection these figures were 83 and 51%, respectively (median 61 months, n = 117) (P < 0.001). The median DFS after local ablation was 9 months, HRQoL was restored within 3 months. Patients after local ablation gained far more QALY's (317) than in the chemotherapy group (165) (P < 0.001). Although overall survival did not reached statistical significance, the median DFS of 9 months suggests a beneficial effect of local tumour ablation for non-resectable colorectal liver metastases. Moreover, compared with systemic chemotherapy more QALY's were gained after local ablative therapy.


Ontogeny of pituitary regulation of growth in the developing rat: comparison of effects of hypophysectomy and hormone replacement on somatic and organ growth, serum insulin-like growth factor-I (IGF-I) and IGF-II levels, and IGF-binding protein levels in the neonatal and juvenile rat.
We investigated pituitary regulation of growth during two critical stages of development in the rat, using hypophysectomy (Hx) with replacement of GH and/or T4. In the neonatal period (Hx on day 6), body weight and tail length were inhibited by 60% and 50%, respectively, while these parameters were inhibited by 80% and 85% by Hx in the juvenile period (Hx on day 45). Administration of T4 alone significantly increased skeletal growth (tail length) in neonatal Hx rats, while T4 alone proved ineffective in promoting somatic growth in juvenile Hx rats. GH effects were greater on body weight than on tail length at both stages of development. Replacement of both GH and T4 restored somatic growth to normal values during both time periods. The brain was the sole organ whose growth appeared to be independent of the pituitary. Hx reduced serum insulin-like growth factor-I (IGF-I) and -II in both age groups, and GH alone restored IGF-I and -II levels to the control range. The major IGF-binding proteins (IGFBPs) were analyzed by Western ligand blots. The effect of Hx on the predominant IGFBP was greater in the juvenile rat. T4 replacement in the neonate and GH replacement in the juvenile rat restored IGFBPs to control levels. We conclude that somatic growth in the rat is less pituitary dependent in the neonatal period. There are also important age-specific differences in organ response to GH and T4. Serum IGFs and their binding proteins are pituitary dependent even in infancy, and GH is their primary regulator. The neonatal Hx rat is an important model for the study of the dynamic development of the pituitary-dependent growth.


Analytical Micromechanics Models for Elastoplastic Behavior of Long Fibrous Composites: A Critical Review and Comparative Study.
Elasto-plastic models for composites can be classified into three categories in terms of a length scale, i.e., macro scale, meso scale, and micro scale (micromechanics) models. In general, a so-called multi-scale model is a combination of those at various length scales with a micromechanics one as the foundation. In this paper, a critical review is made for the elastoplastic models at the micro scale, and a comparative study is carried out on most popular analytical micromechanics models for the elastoplastic behavior of long fibrous composites subjected to a static load, meaning that creep and dynamic response are not concerned. Each model has been developed essentially following three steps, i.e., an elastic homogenization, a rule to define the yielding of a constituent phase, and a linearization for the elastoplastic response. The comparison is made for all of the three aspects. Effects of other issues, such as the stress field fluctuation induced by a high contrast heterogeneity, the stress concentration factors in the matrix, and the different approaches to a plastic Eshelby tensor, are addressed as well. Correlation of the predictions by different models with available experimental data is shown.


Unraveling the Coupling between Conformational Changes and Ligand Binding in Ribose Binding Protein Using Multiscale Molecular Dynamics and Free-Energy Calculations.
Conformational changes of proteins upon ligand binding are usually explained in terms of several mechanisms including the induced fit, conformational selection, or their mixtures. Due to the slow time scales, conventional molecular dynamics (cMD) simulations based on the atomistic models cannot easily simulate the open-to-closed conformational transition in proteins. In our previous study, we have developed an enhanced sampling scheme (generalized replica exchange with solute tempering selected surface charged residues: gREST_SSCR) for multidomain proteins and applied it to ligand-mediated conformational changes in the G134R mutant of ribose-binding protein (RBPG134R) in solution. The free-energy landscape (FEL) of RBPG134R in the presence of a ribose at the binding site included the open and closed states and two intermediates, open-like and closed-like forms. Only the open and open-like forms existed in the FEL without a ribose. In the current study, the coupling between the conformational changes and ligand binding is further investigated using coarse-grained MD, multiple atomistic cMD, and free-energy calculations. The ribose is easily dissociated from the binding site of wild-type RBP and RBPG134R in the cMD simulations starting from the open and open-like forms. In contrast, it is stable at the binding site in the simulations from the closed and closed-like forms. The free-energy calculations provide the binding affinities of different structures, supporting the results of cMD simulations. Importantly, cMD simulations from the closed-like structures reveal transitions toward the closed one in the presence of a bound ribose. On the basis of the computational results, we propose a molecular mechanism in which conformational selection and induced fit happen in the first and second halves of the open-to-closed transition in RBP, respectively.


HIV risk-related sex behaviors among injection drug users, crack smokers, and injection drug users who smoke crack.
This study was designed to assess and compare sex risk behaviors for human immunodeficiency virus (HIV) transmission of three drug user groups: injectors who do not smoke crack, crack smokers who do not inject, and injectors who also smoke crack. Sexual risk behaviors for HIV were assessed among 246 drug users from Denver, Miami, and San Francisco. Respondents were classified into the three drug groups based on self-report and verified through urinalysis and physical inspection. An increased risk for HIV through sexual transmission was associated with crack cocaine use, particularly among those who also injected. Crack smoking injectors were more likely to report sex with an injector, exchanging sex for drugs and/or money, drug use before or during sex, and unprotected sexual intercourse. They also injected more than injectors only, smoked crack as often as smokers only, and reported higher overall frequencies of drug use. These findings, together with the higher rates of gonorrhea and syphilis reported by smokers and injectors/smokers, are indicators of the risk crack poses for the heterosexual transmission of HIV.


Factors in childhood associated with lung function decline to adolescence in cystic fibrosis.
Despite improvements in general health and life expectancy in people with cystic fibrosis (CF), lung function decline continues unabated during adolescence and early adult life. We examined factors present at age 5-years that predicted lung function decline from childhood to adolescence in a longitudinal study of Australasian children with CF followed from 1999 to 2017. Lung function trajectories were calculated for 119 children with CF from childhood (median 5.0 [25%-75%=5.0-5.1]) years) to early adolescence (median 12.5 [25%-75%=11.4-13.8] years). Lung function fell progressively, with mean (standard deviation) annual change -0.105 (0.049) for forced vital capacity (FVC) Z-score (p<0.001), -0.135 (0.048) for forced expiratory volume in 1-second (FEV1) Z-score (p<0.001), -1.277 (0.221) for FEV1/FVC% (p<0.001), and -0.136 (0.052) for forced expiratory flow between 25% and 75% of FVC Z-score (p<0.001). Factors present in childhood predicting lung function decline to adolescence, in multivariable analyses, were hospitalisation for respiratory exacerbations in the first 5-years of life (FEV1/FVC p = 0.001, FEF25-75p = 0.01) and bronchoalveolar lavage neutrophil elastase activity (FEV1/FVC% p = 0.001, FEV1p = 0.05, FEF25-75p = 0.02). No examined factor predicted a decline in the FVC Z-score. Action in the first 5-years of life to prevent and/or treat respiratory exacerbations and counteract neutrophilic inflammation in the lower airways may reduce lung function decline in children with CF, and these should be targets of future research.


Rehydration methods to recover cysticercoids of the rat tapeworm Hymenolepis diminuta from dry flour beetle carcasses.
Terrestrial arthropods host a variety of helminth parasites, yet quantifying the intensity of infection in these hosts post-mortem is challenging because carcasses may desiccate quickly. We recovered cysticercoids of Hymenolepis diminuta from desiccated flour beetle (Tribolium confusum) carcasses by modifying a published insect rehydration procedure. Without rehydration, carcasses dissected more than 1 day post-mortem had noticeable degradation of cysticercoids. Mild rehydration (soaking in water only for 2 days, or 0.5-10% KOH for 1 h followed by 1 day in water, or 0.5% KOH for 1 day) left carcasses tough and time-consuming to dissect, but all parasites could be recovered and were similar in body size to fresh cysticercoids. Moderate rehydration (5-10% KOH for 1 day) allowed all parasites to be recovered and facilitated dissection by partially dissolving internal organs of the beetle while causing little degradation of the cysticercoids. Harsh rehydration (5-10% KOH for 1 day followed by 5 days in water) not only dissolved internal beetle tissues but also severely damaged cysticercoids, such that parasite counts were unreliable. The degree of initial carcass desiccation had little effect on results following rehydration. However, regardless of treatment used, intact cercomers were rarely retained on rehydrated cysticercoids. Rehydration was less successful on early developmental stages of the parasite, which were recovered reliably only as they neared the cysticercoid stage. This method has utility for studies of parasite-induced mortality by permitting accurate and reliable parasite counts from dead, desiccated hosts.


Locus coeruleus stimulation exerts different influences on the dynamic changes of cerebral pial and intraparenchymal vessels.
The present experimental study was undertaken to investigate the effects of locus coeruleus stimulation on the dynamic changes of intraparenchymal vessels and pial vessels. Twelve cats were anaesthetized with alpha-chloralose and urethane. For stimulation of the locus coeruleus, a concentric stainless-steel needle electrode was inserted stereotaxically. During the stimulation, volumetric changes of the intraparenchymal vessels were monitored by a photoelectric method for estimating the cerebral blood volume (CBV) (6 cats), and the diameters of pial arteries were measured continuously using a video camera system (6 cats). The CBV followed a decreasing course during the stimulation of the locus coeruleus. The decrease in CBV from the control value (6.3 vol%) was 0.14 +/- 0.04 vol% at 80 s (p less than 0.05), 0.15 +/- 0.05 vol% at 100 s (p less than 0.05), and 0.15 +/- 0.03 vol% at 120 s (p less than 0.01). After cessation of the stimulation, CBV showed a gradual recovery. On the other hand, the diameters of the pial arteries did not change during or after the stimulation of the locus coeruleus. The above results suggest that the locus coeruleus has a vasoconstrictive effect on the intraparenchymal vessels, although it exerts no apparent influence on the pial arteries.


Consecutively reconstructing absorption and scattering distributions in turbid media with multiple-illumination photoacoustic tomography.
We propose a theoretical framework for consecutively reconstructing absorption and scattering distributions in turbid soft tissue in an iterative manner. This approach takes advantage of the stability of a recently reported least-squares fixed-point iterative method for reconstructing an optical absorption coefficient map to iteratively update estimates of absorption and scattering for each iteration. Simulations demonstrate that this method converges to an accurate estimate of the optical properties within only a small number of iterations and is robust to noise at realistic signal-to-noise levels.


Acoustic Trapping Technique for Studying Calcium Response of a Suspended Breast Cancer Cell: Determination of Its Invasion Potentials.
A noncontact single-beam acoustic trapping technique has proven to be a promising tool for cell manipulation and characterization that provide essential knowledge for a variety of biomedical applications. Here, we investigated cell characteristics as to whether the calcium responses of suspended breast cancer cells to different acoustic trapping forces are related to their invasiveness. For this, we combined a single-beam acoustic trapping system with a 30-MHz press-focused lithium niobate ultrasound transducer and an epifluorescence microscope. Using the system, intracellular calcium changes of suspended MDA-MB-231 (highly invasive) and MCF-7 (weakly invasive) cells were monitored while trapping the cells at different acoustic pressures. The results showed that a single suspended breast cancer cell isolated by the acoustic microbeam behaved differently on the calcium elevation in response to changes in acoustic trapping force, depending on its invasiveness. In particular, the MDA-MB-231 cells exhibited higher calcium elevation than MCF-7 cells when each cell was trapped at low acoustic pressure. Based on these results, we believe that the single-beam acoustic trapping technique has high potential as an alternative tool for determining the degree of invasiveness of suspended breast cancer cells.


Involuntary Neuromuscular Coupling between the Thumb and Finger of Stroke Survivors during Dynamic Movement.
Finger-thumb coordination is crucial to manual dexterity but remains incompletely understood, particularly following neurological injury such as stroke. While being controlled independently, the index finger and thumb especially must work in concert to perform a variety of tasks requiring lateral or palmar pinch. The impact of stroke on this functionally critical sensorimotor control during dynamic tasks has been largely unexplored. In this study, we explored finger-thumb coupling during close-open pinching motions in stroke survivors with chronic hemiparesis. Two types of perturbations were applied randomly to the index with a novel Cable-Actuated Finger Exoskeleton: a sudden joint acceleration stretching muscle groups of the index finger and a sudden increase in impedance in selected index finger joint(s). Electromyographic signals for specific thumb and index finger muscles, thumb tip trajectory, and index finger joint angles were recorded during each trial. Joint angle perturbations invoked reflex responses in the flexor digitorum superficialis (FDS), first dorsal interossei (FDI), and extensor digitorum communis muscles of the index finger and heteronymous reflex responses in flexor pollicis brevis of the thumb (p < 0.017). Phase of movement played a role as a faster peak reflex response was observed in FDI during opening than during closing (p < 0.002) and direction of perturbations resulted in shorter reflex times for FDS and FDI (p < 0.012) for extension perturbations. Surprisingly, when index finger joint impedance was suddenly increased, thumb tip movement was substantially increased, from 2 to 10 cm (p < 0.001). A greater effect was seen during the opening phase (p < 0.044). Thus, involuntary finger-thumb coupling was present during dynamic movement, with perturbation of the index finger impacting thumb activity. The degree of coupling modulated with the phase of motion. These findings reveal a potential mechanism for direct intervention to improve poststroke hand mobility and provide insight on prospective neurologically oriented therapies.


Bright fluorescent Streptococcus pneumoniae for live-cell imaging of host-pathogen interactions.
Streptococcus pneumoniae is a common nasopharyngeal resident in healthy people but, at the same time, one of the major causes of infectious diseases such as pneumonia, meningitis, and sepsis. The shift from commensal to pathogen and its interaction with host cells are poorly understood. One of the major limitations for research on pneumococcal-host interactions is the lack of suitable tools for live-cell imaging. To address this issue, we developed a generally applicable strategy to create genetically stable, highly fluorescent bacteria. Our strategy relies on fusing superfolder green fluorescent protein (GFP) or a far-red fluorescent protein (RFP) to the abundant histone-like protein HlpA. Due to efficient translation and limited cellular diffusion of these fusions, the cells are 25-fold brighter than those of the currently best available imaging S. pneumoniae strain. These novel bright pneumococcal strains are fully virulent, and the GFP reporter can be used for in situ imaging in mouse tissue. We used our reporter strains to study the effect of the polysaccharide capsule, a major pneumococcal virulence factor, on different stages of infection. By dual-color live-cell imaging experiments, we show that unencapsulated pneumococci adhere significantly better to human lung epithelial cells than encapsulated strains, in line with previous data obtained by classical approaches. We also confirm with live-cell imaging that the capsule protects pneumococci from neutrophil phagocytosis, demonstrating the versatility and usability of our reporters. The described imaging tools will pave the way for live-cell imaging of pneumococcal infection and help further understanding of the mechanisms of pneumococcal pathogenesis.


Systems pharmacology of adiposity reveals inhibition of EP300 as a common therapeutic mechanism of caloric restriction and resveratrol for obesity.
Both caloric restriction (CR) and resveratrol (RSV) have beneficial effects on obesity. However, the biochemical pathways that mediate these beneficial effects might be complex and interconnected and have not been fully elucidated. To reveal the common therapeutic mechanism of CR and RSV, we performed a comparative transcriptome analysis of adipose tissues from diet-induced obese (DIO) zebrafish and obese humans. We identified nine genes in DIO zebrafish and seven genes in obese humans whose expressions were regulated by CR and RSV. Although the gene lists did not overlap except for one gene, the gene ontologies enriched in the gene lists were highly overlapped, and included genes involved in adipocyte differentiation, lipid storage and lipid metabolism. Bioinformatic analysis of cis-regulatory sequences of these genes revealed that their transcriptional regulators also overlapped, including EP300, HDAC2, CEBPB, CEBPD, FOXA1, and FOXA2. We also identified 15 and 46 genes that were dysregulated in the adipose tissue of DIO zebrafish and obese humans, respectively. Bioinformatics analysis identified EP300, HDAC2, and CEBPB as common transcriptional regulators for these genes. EP300 is a histone and lysyl acetyltransferase that modulates the function of histone and various proteins including CEBPB, CEBPD, FOXA1, and FOXA2. We demonstrated that adiposity in larval zebrafish was significantly reduced by C646, an inhibitor of EP300 that antagonizes acetyl-CoA. The reduction of adiposity by C646 was not significantly different from that induced by RSV or co-treatment of C646 and RSV. These results indicate that the inhibition of EP300 might be a common therapeutic mechanism between CR and RSV in adipose tissues of obese individuals.


Fluorescence of Picrosirius Red Multiplexed With Immunohistochemistry for the Quantitative Assessment of Collagen in Tissue Sections.
The low cost and simplicity of picrosirius red (PSR) staining have driven its popularity for collagen detection in tissue sections. We extended the versatility of this method by using fluorescent imaging to detect the PSR signal and applying automated quantification tools. We also developed the first PSR protocol that is fully compatible with multiplex immunostaining, making it possible to test whether collagen structure differs across immunohistochemically labeled regions of the tissue landscape. We compared our imaging method with two gold standards in collagen imaging, linear polarized light microscopy and second harmonic generation imaging, and found that it is at least as sensitive and robust to changes in sample orientation. As proof of principle, we used a genetic approach to overexpress beta catenin in a patchy subset of mouse prostate epithelial cells distinguished only by immunolabeling. We showed that collagen fiber length is significantly greater near beta catenin overexpressing cells than near control cells. Our fluorescent PSR imaging method is sensitive, reproducible, and offers a new way to guide region of interest selection for quantifying collagen in tissue sections.