Bacterial killing by dry metallic copper surfaces.
Metallic copper surfaces rapidly and efficiently kill bacteria. Cells exposed to copper surfaces accumulated large amounts of copper ions, and this copper uptake was faster from dry copper than from moist copper. Cells suffered extensive membrane damage within minutes of exposure to dry copper. Further, cells removed from copper showed loss of cell integrity. Acute contact with metallic copper surfaces did not result in increased mutation rates or DNA lesions. These findings are important first steps for revealing the molecular sensitive targets in cells lethally challenged by exposure to copper surfaces and provide a scientific explanation for the use of copper surfaces as antimicrobial agents for supporting public hygiene.


Reminder letter, tailored stepped-care, and self-choice comparison for repeat mammography.
The main benefits of mammography come from regular on-schedule screening. However, few studies have examined interventions to achieve repeat screening. Participants were women aged 50 to 74, recruited through one setting in Rhode Island and another in North Carolina. Participants had a mammogram already scheduled at recruitment, and had to keep that appointment in order to be eligible for the repeat mammography intervention. A total of 1614 women were in the intervention sample. A four-group randomized design was used: Group 1, a simple reminder letter; Group 2, a 2-month, tailored, stepped intervention delivered 2 months after the completed mammogram; Group 3, a 10-month, tailored, stepped intervention delivered 2 months before the repeat mammogram was due; and Group 4, self-choice of one of the above three strategies. The intervention took place between June 1996 and May 1997. The reminder letter and two levels of the stepped intervention were delivered by mail. The third level of the stepped strategy was a counselor telephone call. Groups 2 and 3 were identical, except for timing. Obtaining the next due mammogram within 15 months, based on clinic records. There were no statistically significant differences among the four groups, both in the total sample and at the two sites separately. On average, a simple reminder may be as effective as more complex strategies for women with a prior on-schedule exam. However, attention is still needed to identify women at risk of lapsing from screening. Some women may require more-intensive interventions.


Estrogen receptors and endothelium.
Estrogens, and in particular 17beta-estradiol (E2), play a pivotal role in sexual development and reproduction and are also implicated in a large number of physiological processes, including the cardiovascular system. Both acetylcholine-induced and flow-dependent vasodilation are preserved or potentiated by estrogen treatment in both animal models and humans. Indeed, E2 increases the endothelial production of nitric oxide and prostacyclin and prevents early atheroma through endothelial-mediated mechanisms. Furthermore, whereas it prevents endothelial activation, E2 potentiates the ability of several subpopulations of the circulating or resident immune cells to produce proinflammatory cytokines. The balance between these 2 actions could determine the final effect in a given pathophysiological process. E2 also promotes endothelial healing, as well as angiogenesis. Estrogen actions are essentially mediated by 2 molecular targets: estrogen receptor-alpha (ERalpha) and ERbeta. The analysis of mouse models targeted for ERalpha or ERbeta demonstrated a prominent role of ERalpha in vascular biology. ERalpha directly modulates transcription of target genes through 2 activation functions (AFs), AF-1 and AF-2. Interestingly, an AF-1-deficient ERalpha isoform can be physiologically expressed in the endothelium and appears sufficient to mediate most of the vasculoprotective actions of E2. In contrast, AF-1 is necessary for the E2 actions in reproductive targets. Thus, it appears conceivable to uncouple the vasculoprotective and sexual actions with appropriate selective ER modulators.


Age-dependent differences in the pathogenesis of bovine respiratory syncytial virus infections related to the development of natural immunocompetence.
The severity of respiratory syncytial virus (RSV) infections appears to differ with age in both humans and bovines. A primary RSV infection in naïve infants and in young calves runs a more severe course when they are 1-6 months old than in their first month of life. The relative lack of clinical signs in the first month of age may be due to high levels of maternally derived neutralizing antibodies or low exposure to infectious virus. This study examined whether age-dependent differences in the pathogenesis of bovine RSV (bRSV) between neonatal and young calves may be due to differences in age-dependent immunocompetence. To study the effect of age and immune parameters on bRSV disease in neonatal and young calves, neonatal (1-day-old) calves without maternally derived antibodies were infected experimentally with bRSV and the severity of disease and immune responses were evaluated in comparison with disease in similar 6-week-old infected calves. Neonatal calves had more extensive virus replication and lung consolidation, but lower pro-inflammatory [in particular tumour necrosis factor alpha (TNF-α)] responses, specific humoral immune responses, lung neutrophilic infiltration and clinical signs of disease than 6-week-old calves. The lack of correlation between virus replication and clinical signs suggests an important role of pro-inflammatory cytokines, in particular TNF-α, in the disease. The capacity to produce pro-inflammatory TNF-α appeared to increase with age, and may explain the age-dependent differences in RSV pathogenesis.


Amino Acid Composition of Novel Plant Drinks from Oat, Lentil and Pea.
Plant-based drinks (PBDs) as alternatives to milk is a fast-growing market in much of the western world, with the demand increasing every year. However, most PBDs from a single plant ingredient do not have an amino acid profile that matches human needs. Therefore, this study set out to combine plant ingredients to achieve a more balanced amino acid profile of novel plant drinks, by combining a high content of oat with the pulses pea (Pisum sativum) and lentil (Lens culinaris) in a solution. After removal of the sediment, the resulting plant drinks were composed of what could be kept in suspension. The amino acid and protein composition of the plant drinks were investigated with capillary electrophoresis, to identify the amino acids, and SDS-PAGE to assess the proteins present. The amino acid profile was compared against recommended daily intake (RDI). It was determined that the plant drinks with only oat and lentil did not have a strong amino acid profile, likely due to the higher pH of the lentil concentrate affecting which proteins could be kept in solution. Plant drinks with a combination of both lentil and pea, or only pea, added to the oat drink had an improved concentration of the amino acids that were otherwise in the low end compared to RDI. This includes a high content of phenylalanine, leucine and threonine, as well as a moderate amount of isoleucine, valine and methionine, and a contribution of histidine and lysine. An assessment of stability and sensory parameters was also conducted, concluding there was an advantage of combining oat with a legume, especially pea.


The Life of a Trailing Spouse.
In 1981, I published a paper in the first issue of the Journal of Neuroscience with my postdoctoral mentor, Alan Pearlman. It reported a quantitative analysis of the receptive field properties of neurons in reeler mouse visual cortex and the surprising conclusion that although the neuronal somas were strikingly malpositioned, their receptive fields were unchanged. This suggested that in mouse cortex at least, neuronal circuits have very robust systems in place to ensure the proper formation of connections. This had the unintended consequence of transforming me from an electrophysiologist into a cellular and molecular neuroscientist who studied cell adhesion molecules and the molecular mechanisms they use to regulate axon growth. It took me a surprisingly long time to appreciate that your science is driven by the people around you and by the technologies that are locally available. As a professional puzzler, I like all different kinds of puzzles, but the most fun puzzles involve playing with other puzzlers. This is my story of learning how to find like-minded puzzlers to solve riddles about axon growth and regeneration.


Near-miss crashes and other predictors of motorcycle crashes: Findings from a population-based survey.
Crash and injury surveillance studies have identified a range of rider-related factors, including age, sex, licensure, training and experience, as being associated with motorcycle crash risk. The aim of this study was to establish whether these previously identified factors were associated with crash involvement in an Australian-based population. Data obtained from motorcyclists recruited from road authority licensing offices in a population-based survey design were analyzed. In addition to descriptive analysis, survey logistic regression was used to examine predictors of self-reported motorcycle crashes. A statewide population prevalence study of motorcyclists in New South Wales, Australia, was conducted using a multistage stratified random sampling plan. Participants (n = 503) represented 47% of eligible riders invited to participate. The distribution of responses was weighted to represent the population based on motorcycle registrations as a proxy for active motorcyclists, adjusted for age, sex, and variations in sample size and population density between survey sites. This analysis investigated factors associated with having crashed in the past 12 months. The key predictors of increased crash risk included frequent near-crash experiences (6-10) in the past year (adjusted odds ratio [ORadj] = 5.3; 95% confidence interval [CI], 1.3-21.8), having 4 or more riding demerit points (ORadj = 4.1; 95% CI, 1.1-14.7), and motorcycle type and riding purpose. Sports (ORadj = 2.8; 95% CI, 1.1-7.3) and commuter motorcycles (ORadj = 4.0; 95% CI, 1.1-15.3) were associated with higher odds of crashes compared to cruiser/touring motorcycles. Those whose purpose for riding frequently involved commuting, high-speed roads, or motorcycle sports had higher odds of being involved in a crash compared to riders who rarely took part in such activities. Rider age, license type, and time holding a motorcycle license were not predictive of crash involvement when other factors were taken into account. These findings provide important population-level information and insights about risk exposure for motorcyclists. Taking a more tailored approach to data collection meant that factors associated with crash involvement were identified that are not commonly observed in studies relying on administrative data. In particular, the study highlights the importance of near-crash experiences as warnings to riders and the need to use such experiences as learning opportunities to improve their riding style and safety.


Role of crotoxin in coagulation: novel insights into anticoagulant mechanisms and impairment of inflammation-induced coagulation.
Snake venom phospholipases A2 (svPLA2) are biologically active toxins, capable of triggering and modulating a wide range of biological functions. Among the svPLA2s, crotoxin (CTX) has been in the spotlight of bioprospecting research due to its role in modulating immune response and hemostasis. In the present study, novel anticoagulant mechanisms of CTX, and the modulation of inflammation-induced coagulation were investigated. CTX anticoagulant activity was evaluated using platelet poor plasma (PPP) and whole blood (WB), and also using isolated coagulation factors and complexes. The toxin modulation of procoagulant and pro-inflammatory effects was evaluated using the expression of tissue factor (TF) and cytokines in lipopolysaccharide (LPS)-treated peripheral blood mononuclear cells (PBMC) and in WB. The results showed that CTX impaired clot formation in both PPP and WB, and was responsible for the inhibition of both intrinsic (TF/factor VIIa) and extrinsic (factor IXa/factor VIIIa) tenase complexes, but not for factor Xa and thrombin alone. In addition, the PLA2 mitigated the prothrombinase complex by modulating the coagulation phospholipid role in the complex. In regards to the inflammation-coagulation cross talk, the toxin was capable of reducing the production of the pro-inflammatory cytokines IL-1β, IL-6 and TNF-α, and was followed by decreased levels of TF and procoagulant activity from LPS-treated PBMC either isolated or in WB. The results obtained in the present study recognize the toxin as a novel medicinal candidate to be applied in inflammatory diseases with coagulation disorders.


The aetiology and treatment of varicose pigmentation.
The skin pigmentation of chronic venous insufficiency is associated both with increased melanin production and also with deposition of haemosiderin. Following treatment by compression sclerotherapy this pigmentation usually decreases. Seven patients with varicose pigmentation were assessed prior to treatment by colour photographs and skin biopsies. About one year after sclerotherapy further photographs and biopsies were taken. Six of the patients showed a definite decrease of pigmentation on the photographs. In all seven the amount of melanin had decreased. The results for haemosiderin were less uniform and showed a variable rise or fall. It therefore seems likely that the discoloured legs of chronic venous insufficiency result from stimulation of melanocytes perhaps similar to a sunburn response or to the mechanism in haemosiderosis. In the same way this may be reversible when the stimulus of venous hypertension or ultraviolet radiation is removed. The haemosiderin discolouration may also result from extravasation of red cells through capillaries if blood leaks out of the tissues during injections without adequate pressure bandaging.


Indomethacin but not aspirin increases plasma lithium ion levels.
The pharmacokinetic drug interactions between lithium sulfate and the nonsteroidal anti-inflammatory drugs (NSAIDs) indomethacin and aspirin were studied in ten normal female volunteers restricted to 150-mEq/day of sodium. Indomethacin decreased renal lithium ion elimination by 23% and caused a 40% increase in steady state plasma lithium ion levels. In contrast, aspirin had no effect on plasma lithium ion levels and increased renal lithium ion elimination by only 6%. Renal prostaglandin E2 excretion was suppressed by 50% to 60% of control levels by indomethacin and by 65% to 70% by aspirin. A clinically important drug interaction with the risk of lithium intoxication can occur between lithium salts and indomethacin. However, aspirin did not affect steady state plasma levels of lithium ion and thus may be preferable for antirheumatic treatment of patients undergoing long-term therapy with lithium salts. Frequent monitoring of plasma lithium ion levels is absolutely necessary in patients receiving both lithium salts and NSAIDs.


Gut microbiome comparability of fresh-frozen versus stabilized-frozen samples from hospitalized patients using 16S rRNA gene and shotgun metagenomic sequencing.
Collection of faecal samples for microbiome analysis in acutely sick patients is logistically difficult, particularly if immediate freezing is required (i.e. fresh-frozen, or FF sampling). Previous studies in healthy/non-hospitalized volunteers have shown that chemical stabilization (i.e. stabilized-frozen, or SF sampling) allows room-temperature storage with comparable results to FF samples. To test this in a hospital setting we compared FF and SF approaches across 17 patients undergoing haematopoietic stem cell transplantation (HSCT) using both 16S rRNA gene and shotgun metagenomic sequencing. A paired (same stool specimen) comparison of FF and SF samples was made, with an overall comparable level in relative taxonomic abundances between the two sampling techniques. Though shotgun metagenomic sequencing found significant differences for certain bacterial genera (P < 0.001), these were considered minor methodological effects. Within-sample diversity of either method was not significantly different (Shannon diversity P16SrRNA = 0.68 and Pshotgun = 0.89) and we could not reject the null hypothesis that between-sample variation in FF and SF were equivalent (P16SrRNA = 0.98 and Pshotgun = 1.0). This indicates that SF samples can be used to reliably study the microbiome in acutely sick patient populations, thus creating and enabling further outcomes-based metagenomic studies on similarly valuable cohorts.


Metastatic B-cell lymphoma masquerading as infectious retinitis and vasculitis.
Intraocular lymphoma is a rare ocular malignancy that may occur in the retina or the uvea. Retina or vitreoretinal lymphoma accounts for the majority of cases and is often secondary to diffuse large B-cell lymphoma. In the present study, a 66-year-old Caucasian male with a history of Waldenstrom's macroglobulinemia with diffuse large B-cell lymphoma, presented with blurred vision in the left eye one month following cycle 4 of an R-CHOP regimen. At the time of onset, the patient was being treated for bacterial pneumonia. Visual acuity was 20/25 in his right eye (OD) and 20/30 in the left (OS). Ophthalmologic examination showed intraretinal white infiltrates associated with hemorrhage in the superotemporal midperiphery of the retina and vitritis OS. Initial diagnostic considerations included infectious (cytomegalovirus retinitis, syphilis, toxoplasmosis, tuberculosis), inflammatory (retinal vasculitis associated with autoimmune disease or hypercoagulable states) or malignant (intraocular lymphoma) diseases. The patient did not respond to intravitreal injection of foscarnet and oral valgancyclovir. Systemic work-up and aqueous fluid biopsy were inconclusive. Diagnostic vitrectomy yielded inconclusive results and the patient continued to have progressive loss of vision. A repeat diagnostic vitrectomy with retinal and subretinal biopsy confirmed large B cells consistent with metastatic B-cell lymphoma. A concomitant PET/CT scan was performed that revealed bilateral new pulmonary nodules resulting in additional chemotherapy. Our case shows the diagnostic dilemmas in patients with systemic lymphoma and the possible role of concurrent systemic restaging in patients with ocular complaints, even when in systemic remission.


p values for pathogens: statistical inference from infectious-disease data.
Certain features of infectious-disease data-including the aggregated nature of the data, confounding variables, correlated variables, and non-linear relations-complicate the use of standard statistical procedures. Using data on a helminth infection, we review the use of three parametric tests (analysis of variance, linear regression, and logistic regression), address the complications arising from violation of the assumptions for these tests, and suggest methods of correction. We also compare the relative merits of parametric methods with equivalent non-parametric approaches, and illustrate the differences produced with results from a Kruskal-Wallis test and a t test. The value of using a resampling method-bootstrapping-is also shown. Finally, we discuss problems arising from use of a study design that requires data on the same attribute to be collected from the same individual over a period of time, and present three methods for overcoming this complication, showing that, in the example used, the mixed effect model and generalised estimating equation give similar results.


Lectin histochemistry of malignant fibrohistiocytic tumors.
The staining pattern by Ricinus communis agglutinin (RCA), a lectin used as a good marker for histiocytes, in 24 cases with malignant fibrous histiocytoma (MFH) was studied and compared with that of 12 cases of fibrosarcoma (FS). In 20 (83%) of 24 cases of MFH, varying degrees of RCA binding were observed, whereas only four (33%) of 12 cases of FS were positive. RCA-positive FS included three cases with infantile FS and one adult case with post-radiation FS. Eight adult patients with FS were entirely negative. This positivity rate of RCA binding in MFH was much higher than those of antisera against lysozyme, alpha-1-antitrypsin, and alpha-1-antichymotrypsin previously reported. Seven MFH patients with focal aggregation of RCA-positive benign-appearing (reactive) histiocytes died earlier than the other patients with only scattered RCA-positive histiocytes; 5-year survival rates were 32% and 69%, respectively (p less than 0.05). These findings suggest that RCA reactivity can be used as a potential diagnostic and prognostic marker for MFH.


Chemotherapy of pediatric brain-stem tumors.
Forty-five children harboring brain-stem tumors were treated at the University of California, San Francisco, between 1969 and 1979. Pathological diagnoses were made in 19 patients. All patients received radiation therapy (RT). Thirteen patients received chemotherapy before, during, or immediately after RT. Twenty-four patients were treated with chemotherapy at the time of tumor progression, after initial treatment with RT alone. No statistically significant difference in time to tumor progression or survival was found for treatment with chemotherapy as an adjuvant to RT compared to treatment with RT alone followed by chemotherapy administered at the time of tumor progression. There were, however, more long-term survivors in the group that was first treated with chemotherapy at the time of tumor progression. There was no statistically significant correlation between survival and tumor pathology or location, although there were more long-term survivors among patients harboring low-grade gliomas and among patients with tumors confined to the midbrain. The authors documented the response of some brain-stem tumors to chemotherapy; however, cooperative controlled studies will be required to determine the optimum treatment for this disease.


Factors affecting the decision to defer endotracheal extubation after surgery for congenital heart disease: a prospective observational study.
Fast-tracking and early endotracheal extubation have been described in patients undergoing surgery for congenital heart disease (CHD); however, criteria for patient selection have not been validated in a prospective manner. Our goal in this study was to prospectively identify factors associated with the decision to defer endotracheal extubation in the operating room (OR). We performed a prospective observational study of 275 patients (median age 18 months) at the Mount Sinai Medical Center (MSMC), New York, New York, and 49 patients (median age 25 months) at the University of Tokyo Hospital (UTH), Tokyo, Japan, undergoing surgery for CHD requiring cardiopulmonary bypass. These patients were all eligible for fast-tracking, including extubation in the OR immediately after surgery, according to the respective inclusion/exclusion criteria applied at the 2 sites. Eighty-nine percent of patients at the MSMC, and 65% of patients at the UTH were extubated in the OR. At the MSMC, all patients without aortic cross-clamp, and patients with simple procedures (Risk Adjustment for Congenital Heart Surgery [RACHS] score 1) were extubated in the OR. Among the remaining MSMC patients, regression analysis showed that procedure complexity was still an independent predictor for not proceeding with planned extubation in the OR. Extubation was more likely to be deferred in the RACHS score 3 surgical risk patients compared with the RACHS score 2 group (P = 0.005, odds ratio 3.8 [CI: 1.5, 9.7]). Additionally, trisomy 21 (P = 0.0003, odds ratio 9.9 [CI: 2.9, 34.5]) and age (P = 0.0015) were significant independent predictors for deferring OR extubation. We tested our findings on the patients from the UTH by developing risk categories from the MSMC data that ranked eligible patients according to the chance of OR extubation. The risk categories proved to predict endotracheal extubation in the 49 patients who had undergone surgery at the UTH relative to their overall extubation rate, despite differences in anesthetic regimen and inclusion/exclusion criteria. Preoperatively known factors alone can predict the relative chances of deferring extubation after surgery for CHD. The early extubation strategies applied in the 2 centers were successful in the majority of cases.


Receptor-mediated endocytosis of proteoglycans by human fibroblasts involves recognition of the protein core.
Endocytosis by cultured human skin fibroblasts of 35SO4(2-)-labelled or [3H]leucine-labelled proteoglycans from fibroblast secretions and of 125I-proteodermatan sulphate from pig skin was quantitatively investigated. The following results were obtained. (1) Core proteins prepared by digestion with chondroitin ABC lyase were at least as efficiently endocytosed as native proteoglycans. Pig skin proteodermatan sulphate was a competitive inhibitor of endocytosis of 35SO4(2-)-labelled proteoglycans. (2) Proteoglycans produced in the presence of tunicamycin and native proteoglycans degraded with endoglycosaminidase H were internalized at a normal rate. Several monosaccharides that can be bound by mammalian lectins were unable to influence the internalization of proteoglycans. Treatment of proteoglycans with neuraminidase, however, resulted in an increased clearance rate. (3) Reductive methylation or acetoacetylation of lysine residues was accompanied by a parallel decrease in the rate of proteoglycan endocytosis. Reversal of acetoacetylation normalized the uptake properties. Endocytosis of native proteoglycans was also reduced in the presence of poly-L-lysine, and this reduction in endocytosis was observed as well with proteoglycans synthesized in the presence of the lysine analogue S-2-aminoethylcysteine. These results suggest that the recognition marker required for receptor-mediated endocytosis of proteodermatan sulphate resides in its protein moiety and involves lysine residues.


Coefficient D(av) is more sensitive than fractional anisotropy in monitoring progression of irreversible tissue damage in focal nonactive multiple sclerosis lesions.
Persistently hypointense lesions on T1-weighted MR images have been shown to correlate with the amount of axonal damage and clinical disability in multiple sclerosis (MS) patients. The purpose of this study was to investigate whether diffusion coefficient D(av) and fractional anisotropy (FA) are able to detect quantifiable differences among three groups of focal nonactive multiple sclerosis (MS) lesions that appear qualitatively different on T1-weighted images. Conventional and diffusion tensor MR images of the brain were obtained in 18 patients with relapsing remitting (n = 10) or secondary progressive (n=8) MS and in 18 healthy volunteers. Focal nonactive MS lesions were classified as T1 isointense, T1 mildly hypointense, and T1 severely hypointense on unenhanced T1-weighted images. Differences among groups were assessed with one-way analysis of variance using the T1 lesion appearance as the grouping factor and either FA or coefficient D(av) as the dependent measure. FA was lowest and coefficient D(av) was highest in T1 severely hypointense lesions, and then, in ascending and descending order, respectively, T1 mildly hypointense lesions and T1 isointense lesions. A significant difference in the values of FA was detected only between T1 isointense lesions and T1 severely hypointense lesions (P <.01), whereas no difference was found between T1 mildly hypointense lesions and either one of these two groups. A significant difference was found in the values of coefficient D(av) among all investigated lesion groups. Coefficient D(av) was found to correlate inversely with the T1-weighted contrast ratio (r=-0.58, P <.0001), whereas FA and deltaFA (percentage of FA variation in the lesion, a relative FA measure that minimizes the effect of FA spatial dependence) were not. Coefficient D(av) is more sensitive than FA to variations in the degree of T1 hypointensity and, thus, in the amount of the permanent brain tissue damage in patients with MS.


[Psychoeducation of patients and their family members during episode psychosis].
The concept of psychoeducation is close to the concept of therapeutic education and refers to a kind of education intervention targeting people with a mental health condition. In the framework of psychosis, psychoeducation can be offered to patients, family members or both. The efficacy of patient psychoeducation on treatment adherence or social functioning is well-established but only if the family benefits of a joint psychoeducational intervention. Family psychoeducation, even without patient psychoeducation has proven efficacy in reducing relapse rate. This reduction is of the same order of magnitude as that obtained with an antipsychotic medication.


A missing enzyme in thiamin thiazole biosynthesis: identification of TenI as a thiazole tautomerase.
In many bacteria tenI is found clustered with genes involved in thiamin thiazole biosynthesis. However, while TenI shows high sequence similarity with thiamin phosphate synthase, the purified protein has no thiamin phosphate synthase activity, and the role of this enzyme in thiamin biosynthesis remains unknown. In this contribution, we identify the function of TenI as a thiazole tautomerase, describe the structure of the enzyme complexed with its reaction product, identify the substrates phosphate and histidine 122 as the acid/base residues involved in catalysis, and propose a mechanism for the reaction. The identification of the function of TenI completes the identification of all of the enzymes needed for thiamin biosynthesis by the major bacterial pathway.


Characterization of osteolytic, osteoblastic, and mixed lesions in a prostate cancer mouse model using 18F-FDG and 18F-fluoride PET/CT.
The combination of small-animal PET/CT scans and conventional imaging methods may enhance the evaluation of in vivo biologic interactions of murine models in the study of prostate cancer metastasis to bone. Small-animal PET/CT scans using (18)F-fluoride ion and (18)F-FDG coregistered with high-resolution small-animal CT scans were used to longitudinally assess the formation of osteoblastic, osteolytic, and mixed lesions formed by human prostate cancer cell lines in a severe combined immunodeficient (SCID) mouse tibial injection model. These scans were correlated with plain radiographs, histomorphometry, and soft-tissue measurements. Small-animal PET/CT scans were able to detect biologic activity of cells that induced an osteoblastic lesion 2 wk earlier than on plain radiographs. Furthermore, both the size and the activity of the lesions detected on PET/CT images significantly increased at each successive time point (P < 0.05). (18)F-FDG lesions strongly correlated with soft-tissue measurements, whereas (18)F-fluoride ion activity correlated with bone volume measured on histomorphometric analysis (P < 0.005). Osteolytic lesions were successfully quantified using small-animal CT, whereas lesion sizes measured on (18)F-FDG PET scans also strongly correlated with soft-tissue tumor burden (P < 0.05). In contrast, for mixed lesions, (18)F-fluoride ion and (18)F-FDG PET/CT scans detected only minimal activity. (18)F-FDG and (18)F-fluoride ion PET/CT scans can be useful tools in characterizing pure osteolytic and osteoblastic lesions induced by human prostate cancer cell lines. The value of this technology needs further evaluation to determine whether these studies can be used effectively to detect more subtle responses to different treatment regimens in animal models.


Cohort profile: the Olmsted County hypertensive disorders of pregnancy (HDP) cohort using the Rochester Epidemiology Project.
The Olmsted County hypertensive disorders of pregnancy (HDP) cohort is a population-based retrospective study designed to compare the incidence of HDP on a per-pregnancy and per-woman basis and to identify associations between HDP with ageing-related diseases, as well as accumulation of multimorbidity. Using the Rochester Epidemiology Project (REP) medical records-linkage system, a cohort was collected consisting of women who gave birth in Olmsted County between 1976 and 1982. After exclusions, a per-pregnancy cohort of 7544 women with 9862 pregnancies between 1976 and 1982 was identified, and their delivery information was manually reviewed. A subset of these women comprised the per-woman cohort of 4322 pregnancies from 1839 women with delivery information available throughout the entirety of their childbearing years, along with decades of follow-up data available for research via the REP. By constructing both per-pregnancy and per-woman cohorts, we reported a doubling of HDP incidence rates when assessed on a per-woman basis compared with rates observed on a per-pregnancy basis. Moreover, in addition to finding that women with a history of HDP developed specific diseases at higher rates and at early ages, we also discovered that a history of HDP is associated with accelerated ageing, through accumulation of multimorbidity. In addition to these outcomes described above, many other potential outcomes of interest for studies of HDP can be ascertained from accessing the electronic health records (EHR) and billing systems available through the REP. These data can include all International Classification of Diseases (ICD)-9 and ICD-10 and Current Procedural Terminology coded diagnoses and procedures, healthcare utilisation, including office visits, hospitalisations and emergency room visits, and full text of the EHR that is available for chart abstraction or for natural language processing of the clinical notes.


Jones matrix analysis of dichroic phase retarders realized in soft matter composite materials.
Materials showing birefringence and polarization selective absorption (dichroism) affect the polarization state of incoming light in a peculiar way, quite different from the one exhibited by phase retarders like waveplates. In this paper, we report on the characterization of a Polymer LIquid CRYstal Polymer Slices (POLICRYPS) diffraction grating used as a dichroic phase retarder; the dichroic behaviour of the grating is due to the polarization-dependent diffraction efficiency. Experimental data are validated with a theoretical model based on the Jones matrix formalism, while the grating behavior is modeled by means of the dichroic matrix. In this way, the birefringence of the analyzed structure is easily obtained. For comparison purposes, also two systems different from POLICRYPS have been fabricated and tested.


Evolutionary convergence on sleep loss in cavefish populations.
Patterns of sleep vary widely among species, but the functional and evolutionary principles responsible for this diversity remain unknown. The characin fish, Astyanax mexicanus, has eyed surface and numerous blind cave populations. The cave populations are largely independent in their origins, and the species is ideal for studying the genetic bases of convergent evolution. Here we show that this system is also uniquely valuable for the investigation of variability in patterns of sleep. We find that a clearly defined change in ecological conditions, from surface to cave, is correlated with a dramatic reduction in sleep in three independently derived cave populations of A. mexicanus. Analyses of surface × cave hybrids show that the alleles for reduced sleep in the Pachón and Tinaja cave populations are dominant in effect to the surface alleles. Genetic analysis of hybrids between surface and Pachón cavefish suggests that only a small number of loci with dominant effects are involved. Our results demonstrate that sleep is an evolutionarily labile phenotype, highly responsive to changes in ecological conditions. To our knowledge, this is the first example of a single species with a convergence on sleep loss exhibited by several independently evolved populations correlated with population-specific ecologies.


[Comparative biological value of low-protein diets with individual amino acid supplements].
The biological value of low-protein rations was distinctly increased following their enrichment in lysine, methionine, cystine, sodium and potassium glutamate, both individually and in a mixture. The additves under study helped mitigate destruction of the bone marrow cells and stabilize factors reflecting the immunobiological reactivity for the organism.


Higher adalimumab maintenance regimen is more effective than standard dose in anti-TNF experienced Crohn's disease patients.
Many Crohn's disease patients treated with anti-tumor necrosis factor (TNF) therapies suffer from loss of response over time and require dose escalation. The aim of this study was to evaluate the efficacy and safety of treating anti-TNF experienced Crohn's disease patients with higher maintenance regimens of adalimumab. In a retrospective observational study, Crohn's disease patients receiving adalimumab were categorized according to their maintenance regimen; 40 mg weekly, 80 mg every other week or greater were defined as a high-dose maintenance regimen and 40 mg every other week was defined as a standard maintenance regimen. The primary outcome was time to treatment failure. Thirty-nine patients were started on high-dose regimens following induction and 40 patients received the standard regimen. According to a Kaplan-Meier survival curve analysis, time to treatment failure was significantly longer in patients in the high-dose group (P = 0.0015). Patients on high-dose adalimumab had a lower treatment failure rate (hazard ratio 0.21; P = 0.0005) when compared to patients on the standard regimen, after adjusting for induction dose and concomitant immunomodulator use. No difference in adverse events was identified between the groups (31 vs. 30%; P = 0.94). High-dose maintenance regimens were more effective than the standard adalimumab maintenance protocol with better short and long-term clinical outcomes.


Therapy of murine pulmonary aspergillosis with antibody-alliinase conjugates and alliin.
Aspergillus fumigatus is an opportunistic fungal pathogen responsible for invasive aspergillosis in immunocompromised individuals. The high morbidity and mortality rates as well as the poor efficacy of antifungal agents remain major clinical concerns. Allicin (diallyl-dithiosulfinate), which is produced by the garlic enzyme alliinase from the harmless substrate alliin, has been shown to have wide-range antifungal specificity. A monoclonal antibody (MAb) against A. fumigatus was produced and chemically ligated to the enzyme alliinase. The purified antibody-alliinase conjugate bound to conidia and hyphae of A. fumigatus at nanomolar concentrations. In the presence of alliin, the conjugate produced cytotoxic allicin molecules, which killed the fungus. In vivo testing of the therapeutical potential of the conjugate was carried out in immunosuppressed mice infected intranasally with conidia of A. fumigatus. Intratracheal (i.t.) instillation of the conjugate and alliin (four treatments) resulted in 80 to 85% animal survival (36 days), with almost complete fungal clearance. Repetitive intratracheal administration of the conjugate and alliin was also effective when treatments were initiated at a more advanced stage of infection (50 h). The fungi were killed specifically without causing damage to the lung tissue or overt discomfort to the animals. Intratracheal instillation of the conjugate without alliin or of the unconjugated monoclonal antibody significantly delayed the death of the infected mice, but only 20% of the animals survived. A limitation of this study is that the demonstration was achieved in a constrained setting. Other routes of drug delivery will be investigated for the treatment of pulmonary and extrapulmonary aspergillosis.


Protective effect of the ethyl acetate fraction of Sargassum muticum against ultraviolet B-irradiated damage in human keratinocytes.
The aim of this study was to investigate the cytoprotective properties of the ethyl acetate fraction of Sargassum muticum (SME) against ultraviolet B (UVB)-induced cell damage in human keratinocytes (HaCaT cells). SME exhibited scavenging activity toward the 1,1-diphenyl-2-picrylhydrazyl radicals and hydrogen peroxide (H(2)O(2)) and UVB-induced intracellular reactive oxygen species (ROS). SME also scavenged the hydroxyl radicals generated by the Fenton reaction (FeSO(4) + H(2)O(2)), which was detected using electron spin resonance spectrometry. In addition, SME decreased the level of lipid peroxidation that was increased by UVB radiation, and restored the level of protein expression and the activities of antioxidant enzymes that were decreased by UVB radiation. Furthermore, SME reduced UVB-induced apoptosis as shown by decreased DNA fragmentation and numbers of apoptotic bodies. These results suggest that SME protects human keratinocytes against UVB-induced oxidative stress by enhancing antioxidant activity in cells, thereby inhibiting apoptosis.


Fatalism and Suicidal Behaviors Among Chinese College Students: The Mediating Role of Depressive Symptoms and the Moderating Role of Coping Strategies.
The aim of this study was to investigate the relationship between fatalism and suicidal behaviors, the mediating role of depressive symptoms, and the moderating effect of coping strategies on the mediating process. A total of 519 participants completed the Multidimensional Fatalism Scale for General Life Events, the Center for Epidemiologic Studies-Depression scale, the Simplified Coping Style Questionnaire, and the Suicidal Behaviors Questionnaire-Revised. Results suggest that depressive symptoms partially mediated the relationship between fatalism and suicidal behaviors. Active coping moderated the mediating effect of depressive symptoms. The higher the active coping level, the weaker the mediating effect. The findings revealed that the mechanism of fatalism affecting suicidal behaviors, and had theoretical and empirical value for the prevention and intervention of suicide among college students.


Clinical Reasoning and Diagnostic Error: A Call to Merge Two Worlds to Improve Patient Care.
Numerous and substantial challenges exist in the provision of safe, cost-effective, and efficient health care. The prevalence and consequences of diagnostic error, one of these challenges, have been established by the literature; however, these errors persist, and the pace of improvement has been slow. One potential reason for the lack of needed progress is that addressing delayed and wrong diagnoses will require contributions from 2 currently distinct worlds: clinical reasoning and diagnostic error. In this Invited Commentary, the authors argue for merging the diagnostic error and clinical reasoning fields as the perspectives, frameworks, and methodologies of these 2 fields could be leveraged to yield a more aligned approach to understanding and subsequently to mitigating diagnostic error. The authors focus on the problem of diagnostic labeling (a categorization task where one has to choose the correct label or diagnosis). The authors elaborate on why this alignment could help guide health care improvement efforts, using the vexing problem of context specificity that leads to unwanted variance in health care as an example.


Cadaveric verification of the Eclipse AAA algorithm for spine SBRT treatments with titanium hardware.
To assess the accuracy of the Eclipse Analytical Anisotropic Algorithm when calculating dose for spine stereotactic body radiation therapy treatments involving surgically implanted titanium hardware. A human spine was removed from a cadaver, cut sagittally along the midline, and then separated into thoracic and lumbar sections. The thoracic section was implanted with titanium stabilization hardware; the lumbar section was not implanted. Spine sections were secured in a water phantom and simulated for treatment planning using both standard and extended computed tomography (CT) scales. Target volumes were created on both spine sections. Dose calculations were performed using (1) the standard CT scale with relative electron density (RED) override of image artifacts and hardware, (2) the extended CT scale with RED override of image artifacts only, and (3) the standard CT scale with no RED overrides for hardware or artifacts. Plans were delivered with volumetric modulated arc therapy using a 6-MV beam with and without a flattening filter. A total of 3 measurements for each plan were made with Gafchromic film placed between the spine sections and compared with Eclipse dose calculations using gamma analysis with a 2%/2 mm passing criteria. A single measurement in a homogeneous phantom was made for each plan before actual delivery. Gamma passing rates for measurements in the homogeneous phantom were 99.6% or greater. Passing rates for measurements made in the lumbar spine section without hardware were 99.3% or greater; measurements made in the thoracic spine containing titanium were 98.6 to 99.5%. Eclipse Analytical Anisotropic Algorithm can adequately model the effects of titanium implants for spine stereotactic body radiation therapy treatments using volumetric modulated arc therapy. Calculations with standard or extended CT scales give similarly accurate results.


Effects of tumor metabolic microenvironment on regulatory T cells.
Recent studies have shown that on one hand, tumors need to obtain a sufficient energy supply, and on the other hand they must evade the body's immune surveillance. Because of their metabolic reprogramming characteristics, tumors can modify the physicochemical properties of the microenvironment, which in turn affects the biological characteristics of the cells infiltrating them. Regulatory T cells (Tregs) are a subset of T cells that regulate immune responses in the body. They exist in large quantities in the tumor microenvironment and exert immunosuppressive effects. The main effect of tumor microenvironment on Tregs is to promote their differentiation, proliferation, secretion of immunosuppressive factors, and chemotactic recruitment to play a role in immunosuppression in tumor tissues. This review focuses on cell metabolism reprogramming and the most significant features of the tumor microenvironment relative to the functional effects on Tregs, highlighting our understanding of the mechanisms of tumor immune evasion and providing new directions for tumor immunotherapy.


Cortico-Striatal Cross-Frequency Coupling and Gamma Genesis Disruptions in Huntington's Disease Mouse and Computational Models.
Abnormal gamma band power across cortex and striatum is an important phenotype of Huntington's disease (HD) in both patients and animal models, but neither the origin nor the functional relevance of this phenotype is well understood. Here, we analyzed local field potential (LFP) activity in freely behaving, symptomatic R6/2 and Q175 mouse models and corresponding wild-type (WT) controls. We focused on periods of quiet rest, which show strong γ activity in HD mice. Simultaneous recording from motor cortex and its target area in dorsal striatum in the R6/2 model revealed exaggerated functional coupling over that observed in WT between the phase of delta frequencies (1-4 Hz) in cortex and striatum and striatal amplitude modulation of low γ frequencies (25-55 Hz; i.e., phase-amplitude coupling, PAC), but no evidence that abnormal cortical activity alone can account for the increase in striatal γ power. Both HD mouse models had stronger coupling of γ amplitude to δ phase and more unimodal phase distributions than their WT counterparts. To assess the possible role of striatal fast-spiking interneurons (FSIs) in these phenomena, we developed a computational model based on additional striatal recordings from Q175 mice. Changes in peak γ frequency and power ratio were readily reproduced by our computational model, accounting for several experimental findings reported in the literature. Our results suggest that HD is characterized by both a reorganization of cortico-striatal drive and specific population changes related to intrastriatal synaptic coupling.


Youths and HIV/AIDS: psychiatry's role in a changing epidemic.
To review the past 10 years of published research on human immunodeficiency virus/acquired immunodeficiency syndrome (HIV/AIDS) in the United States, including psychosocial and psychiatric risk factors, epidemiology, biology, neurocognitive and psychiatric sequelae, disclosure issues, prevention strategies, and biological and behavioral treatments. Researchers reviewed the English-language literature with a focus on child and adolescent risk factors associated with HIV/AIDS, prevention, and treatment. Substantial scientific advances have occurred over the past two decades leading to decreased morbidity and mortality in the United States from AIDS-related opportunistic infections. At the same time, rates of HIV infection are increasing in teenagers, young women, and minorities, and growing numbers of youths are living with an infected family member. Understanding HIV risk behavior requires a broad theoretical framework. Comprehensive HIV prevention programs have led to reduced risk behavior among HIV-affected youths and teens at risk of infection. Biological and behavioral treatments of HIV infection continue to evolve and have led to longer life span, improved quality of life, and fewer psychiatric problems. HIV/AIDS has significant mental health implications, and psychiatry can play a critical role in curbing the epidemic. With minimal effort, mental health professionals can adapt and apply the strategies that they use to treat psychiatric symptoms to prevent HIV transmission behaviors.


Too sweet: cheminformatics for deglycosylation in natural products.
Sugar units in natural products are pharmacokinetically important but often redundant and therefore obstructing the study of the structure and function of the aglycon. Therefore, it is recommended to remove the sugars before a theoretical or experimental study of a molecule. Deglycogenases, enzymes that specialized in sugar removal from small molecules, are often used in laboratories to perform this task. However, there is no standardized computational procedure to perform this task in silico. In this work, we present a systematic approach for in silico removal of ring and linear sugars from molecular structures. Particular attention is given to molecules of biological origin and to their structural specificities. This approach is made available in two forms, through a free and open web application and as standalone open-source software.


The impact of biological sex on the response to noise and otoprotective therapies against acoustic injury in mice.
Noise-induced hearing loss (NIHL) is the most prevalent form of acquired hearing loss and affects about 40 million US adults. Among the suggested therapeutics tested in rodents, suberoylanilide hydroxamic acid (SAHA) has been shown to be otoprotective from NIHL; however, these results were limited to male mice. Here we tested the effect of SAHA on the hearing of 10-week-old B6CBAF1/J mice of both sexes, which were exposed to 2 h of octave-band noise (101 dB SPL centered at 11.3 kHz). Hearing was assessed by measuring auditory brainstem responses (ABR) at 8, 16, 24, and 32 kHz, 1 week before, as well as at 24 h and 15-21 days following exposure (baseline, compound threshold shift (CTS) and permanent threshold shift (PTS), respectively), followed by histologic analyses. We found significant differences in the CTS and PTS of the control (vehicle injected) mice to noise, where females had a significantly smaller CTS at 16 and 24 kHz (p < 0.0001) and PTS at 16, 24, and 32 kHz (16 and 24 kHz p < 0.001, 32 kHz p < 0.01). This sexual dimorphic effect could not be explained by a differential loss of sensory cells or synapses but was reflected in the amplitude and amplitude progression of wave I of the ABR, which correlates with outer hair cell (OHC) function. Finally, the frequency of the protective effect of SAHA differed significantly between males (PTS, 24 kHz, p = 0.002) and females (PTS, 16 kHz, p = 0.003), and the magnitude of the protection was smaller in females than in males. Importantly, the magnitude of the protection by SAHA was smaller than the effect of sex as a biological factor in the vehicle-injected mice. These results indicate that female mice are significantly protected from NIHL in comparison to males and that therapeutics for NIHL may have a different effect in males and females. The data highlight the importance of analyzing NIHL experiments from males and females, separately. Finally, these data also raise the possibility of effectors in the estrogen signaling pathway as novel therapeutics for NIHL.


Intravitreal injection of ranibizumab for foveal-macular pattern dystrophy: case report.
In the recent years, anti-angiogenic medications have successfully treated other diseases associated with choroidal neovascularization. The anti-angiogenic therapy alone or combined with LASER and/or steroids has been effective in controlling ocular neovascularization, not only restricted to the treatment of typical membranes due to macular degeneration in the wet form. The discovery and subsequent use of these drugs has revolutionized medicine and ophthalmology. This report illustrates an example of successful treatment in a challenging pathology where it was found important visual and anatomical response after the use of ranibizumab.


Heterogeneity of genetic changes associated with acquired crizotinib resistance in ALK-rearranged lung cancer.
Anaplastic lymphoma kinase (ALK)-rearranged non-small-cell lung cancer (NSCLC) is markedly sensitive to the ALK inhibitor crizotinib. However, acquired resistance to crizotinib is inevitable through several mechanisms. Therefore, this study was conducted to identify genetic alterations associated with crizotinib resistance. Tumor samples were derived from seven ALK-positive NSCLC patients who showed acquired resistance to crizotinib, and these patients were analyzed for ALK, EGFR, and KRAS mutations and ALK and EGFR gene amplifications. In vitro cytotoxicity of crizotinib and ALK downstream signals were compared between crizotinib-naive and -resistant NSCLC cells. After a median duration of 6 months (range, 4-12 months), seven ALK-positive NSCLC patients developed acquired resistance to crizotinib. Three patients harbored secondary ALK mutations, including one patient with both mutations: L1196M (n = 2) and G1269A (n = 2). Of note, one patient displayed ALK gene copy number gain (4.1-fold increase compared with the pre-crizotinib specimen) and EGFR L858R mutation with high polysomy. The amphiregulin concentration was high in the supernatant fluid from five patients with malignant pleural effusion (116.4-18934.0 pg/ml). SNU-2535 cells derived from a patient who harbored the G1269 mutation were resistant to crizotinib treatment similar to H3122 CR1 cells. L1196M and G1269A mutant clones were less sensitive to crizotinib and ALK downstream signals were ineffectively suppressed in these clones. Genetic changes associated with crizotinib resistance are heterogeneous in ALK-rearranged NSCLC patients who respond to crizotinib and subsequently develop resistance.


GSK3β regulates physiological migration of stem/progenitor cells via cytoskeletal rearrangement.
Regulation of hematopoietic stem and progenitor cell (HSPC) steady-state egress from the bone marrow (BM) to the circulation is poorly understood. While glycogen synthase kinase-3β (GSK3β) is known to participate in HSPC proliferation, we revealed an unexpected role in the preferential regulation of CXCL12-induced migration and steady-state egress of murine HSPCs, including long-term repopulating HSCs, over mature leukocytes. HSPC egress, regulated by circadian rhythms of CXCL12 and CXCR4 levels, correlated with dynamic expression of GSK3β in the BM. Nevertheless, GSK3β signaling was CXCL12/CXCR4 independent, suggesting that synchronization of both pathways is required for HSPC motility. Chemotaxis of HSPCs expressing higher levels of GSK3β compared with mature cells was selectively enhanced by stem cell factor-induced activation of GSK3β. Moreover, HSPC motility was regulated by norepinephrine and insulin-like growth factor-1 (IGF-1), which increased or reduced, respectively, GSK3β expression in BM HSPCs and their subsequent egress. Mechanistically, GSK3β signaling promoted preferential HSPC migration by regulating actin rearrangement and microtubuli turnover, including CXCL12-induced actin polarization and polymerization. Our study identifies a previously unknown role for GSK3β in physiological HSPC motility, dictating an active, rather than a passive, nature for homeostatic egress from the BM reservoir to the blood circulation.


[Mechanism of impairment of calcium metabolism caused by toxin T-2; the role of the vitamin D-dependent endocrine system].
Functions of the vitamin D-dependent endocrine system were studied in rats deprived of the vitamin after administration of T-2 toxin. Impairments of calcium metabolism, alterations in the enzymatic activity related to vitamin D3 bioactivation and receptor binding of the hormonal form 1,25(OH)2D3 were similar both in rats deprived of vitamin D, treated with T-2 toxin within 5 days at a dose of 0.54 mg/kg and in the corresponding controls. At the same time, reduction of the calcium metabolism patterns was retarded in rats obtaining vitamin D3 simultaneously with T-2 toxin. This effect was expressed as a decrease in normalization of 25-OHD concentration in blood, absence of renal I-hydroxylase 25-OHD3 activation, inspite of the higher content of parath hormone in blood and of cAMP in kidney, while concentration of bound 1,25(OH)2D3 receptors was distinctly decreased in tissues-targets in vivo. Thus, the effects of T-2 toxin on the vitamin D-dependent endocrine system were manifested as development of the vitamin, secondary deficiency, as resistance of I-hydroxylase 25OHD3 to regulating effect of parath hormone as well as inhibition of interaction between the complexes 1,25(OH)2D3-receptor and chromatin.


Risk Factors and Comorbidities for Onychomycosis: Implications for Treatment with Topical Therapy.
A number of comorbidities and risk factors complicate the successful management of onychomycosis. Underlying conditions and patient characteristics, such as tinea pedis, age, and obesity, contribute to risk, whereas comorbidities, such as diabetes and psoriasis, can increase susceptibility to the disease. There are limited data on treatment effectiveness in these patients. Here, the authors review post hoc analyses of efinaconazole topical solution, 10%, in mild-to-moderate onychomycosis and present new data in terms of age and obesity. The only post hoc analysis to report significant differences so far is gender, where female patients do much better; however, the reasons are unclear. The authors report significant differences in terms of efficacy in obese patients who do not respond as well as those with normal body mass index (P=0.05) and in patients who have their co-existing tinea pedis treated compared to those in whom co-existing tinea pedis was not treated (P=0.025). Although there is a trend to reduced efficacy in older patients and those with co-existing diabetes, differences were not significant. More research is needed in onychomycosis patients with these important risk factors and comorbidities to fully evaluate the treatment challengse and possible solutions.


Babesiosis.
Babesiosis is an emerging, tick-transmitted, zoonotic disease caused by hematotropic parasites of the genus Babesia. Babesial parasites (and those of the closely related genus Theileria) are some of the most ubiquitous and widespread blood parasites in the world, second only to the trypanosomes, and consequently have considerable worldwide economic, medical, and veterinary impact. The parasites are intraerythrocytic and are commonly called piroplasms due to the pear-shaped forms found within infected red blood cells. The piroplasms are transmitted by ixodid ticks and are capable of infecting a wide variety of vertebrate hosts which are competent in maintaining the transmission cycle. Studies involving animal hosts other than humans have contributed significantly to our understanding of the disease process, including possible pathogenic mechanisms of the parasite and immunological responses of the host. To date, there are several species of Babesia that can infect humans, Babesia microti being the most prevalent. Infections with Babesia species generally follow regional distributions; cases in the United States are caused primarily by B. microti, whereas cases in Europe are usually caused by Babesia divergens. The spectrum of disease manifestation is broad, ranging from a silent infection to a fulminant, malaria-like disease, resulting in severe hemolysis and occasionally in death. Recent advances have resulted in the development of several diagnostic tests which have increased the level of sensitivity in detection, thereby facilitating diagnosis, expediting appropriate patient management, and resulting in a more accurate epidemiological description.


Arcanobacterium hippocoleae sp. nov., from the vagina of a horse.
A polyphasic taxonomic study was performed on a previously unidentified gram-positive, facultatively anaerobic, diphtheroid-shaped organism isolated from a vaginal discharge of a horse. Comparative 16S rRNA gene sequencing demonstrated that the strain was a member of the genus Arcanobacterium, but sequence divergence values of >4% with described species of this genus (viz: Arcanobacterium haemolyticum, Arcanobacterium bernardiae, Arcanobacterium phocae, Arcanobacterium pluranimalium and Arcanobacterium pyogenes) demonstrated that the isolate represented a novel species. The unknown bacterium was readily distinguished from other Arcanobacterium species by biochemical tests. Based on phylogenetic and phenotypic evidence, it is proposed that the unknown bacterium be classified as Arcanobacterium hippocoleae sp. nov. The type strain of A. hippocoleae is CCUG 44697T (= CIP 106850T).


Clinical significance of vestibular evoked myogenic potentials in benign paroxysmal positional vertigo.
To investigate the vestibular evoked myogenic potentials (VEMPs) resulting in benign paroxysmal positional vertigo (BPPV) patients and to verify its clinical applications in BPPV. A prospective study. Tertiary referral dizziness center. Forty-one patients with diagnosis of BPPV and 92 healthy volunteers who underwent VEMP testing. Patients were treated by canalith repositioning maneuvers according to the affected canal, and testing of VEMP was performed at diagnosis and after treatment. Testing of VEMP was performed in BPPV patients and in the control group. The number of times the canalith repositioning maneuver was repeated until the patient's report of relief from vertigo and findings of negative positioning test were recorded to find out the relationship between VEMP results and the progress of disease. Vestibular evoked myogenic potential results of BPPV patients showed prolonged p13 and n23 latencies compared with those of the control group, and we could not find any significant difference in VEMP latencies between patients with posterior and horizontal canal type of BPPV. The number of times that the maneuver was repeated did not correlate with the degree of latency prolongation, but in the "no response" group, the number of times was considerably greater than that in the "response" group. We found that VEMP latencies are increased in BPPV patients, which may signify neuronal degenerative changes in the macula of the saccule. When an extensive neuronal damage was suspected by VEMP results such as "no response" in VEMP, the disease progress showed a chronic and resistive course. Therefore, we propose that VEMP could be a useful method to determine a clinical prognosis of patients with BPPV.


The efficacy and safety of early initiation of preoperative analgesia with celecoxib in patients underwent arthroscopic knee surgery: A randomized, controlled study.
This study aimed to investigate the efficacy and safety of celecoxib 24 hours preoperative, 1 hour preoperative, and 4 hours postoperative administration in patients with arthroscopic knee surgery (AKS).In all, 206 patients who underwent AKS were consecutively recruited and randomized into 3 groups: (1) early preoperative analgesia group (EPEA), celecoxib 400 mg 24 hours preoperative administration; (2) preoperative analgesia group (PEA), celecoxib 400 mg 1 hour preoperative administration; (3) postoperative analgesia group (POA), celecoxib 400 mg 4 hours postoperative administration. Pain visual analog scale (VAS) scores (at rest and at 90 flexion) and patient global assessment (PGA) score were evaluated before and after operation, and also pethidine consumption and adverse events (AEs).The pain-rest VAS score, percentage of patients with moderate-severe pain at rest, and PGA score in the EPEA and PEA groups were decreased compared with POA group at 8 and 12 hours postoperation. Besides, pain-flexion to 90 VAS score in EPEA and PEA groups were also reduced compared with POA group at 8 hours postsurgery. Interestingly, the percentage of patients with moderate-severe pain at 90 flexion at 8 hours postsurgery in PEA group was fewer compared with POA group, whereas at 4 hours postoperation it was reduced in EPEA group compared with PEA and POA groups. As to consumption of pethidine, it was numerically decreased in EPEA and PEA groups compared with POA group. No difference between each 2 groups was found in AEs.Celecoxib was effective and safe as pre-emptive analgesia in AKS, and 1 hour administration before operation might be an optimal choice.


Prevalence of food-poisoning (enterotoxigenic) Clostridium perfringens type A in blood and fish meal.
Four of 9 strains of Cl. perfringens type A isolated from 4 of 7 blood meal and 5 of 12 fish meal samples were found to be enterotoxigenic on biological tests and immunodiffusion test with enterotoxin specific serum. Enterotoxin produced by these strains was identical with that of food-poisoning strains. Of these 4 strains, 3 were heat-resistant and 1 was heat-sensitive spore formers. Rate of sporulation of enterotoxigenic strains was greater than the non-enterotoxigenic ones. Animal and public health significance of these strains were discussed.


Development of an iron overload HepG2 cell model using ferrous ammonium citrate.
Cell-based iron overload models provide tremendous utility for the investigations into the pathogenesis of different diseases as well as assessing efficacy of various therapeutic strategies. In the literature, establishing such models vary widely with regards to cell lines, iron source, iron treatment conditions and duration. Due to this diversity, researchers reported significant differences in the measured outcomes, either in cellular function or response to a stimulus. Herein, we report the process required to establish an iron overload HepG2 cell model to achieve a consistent and reproducible results such that the literature can strive towards a consensus. Iron loading in cells was achieved with 50 μM of iron every 24 h for 2 days, followed by an additional 24 h of maintenance in fresh media. We demonstrated that iron overloaded cells had significantly increased ROS generation, labile and total iron whilst having various cellular functions resemble cells without iron overload. The present report addresses key pitfalls with regards to the lack of consensus currently present in the literature.


Immune defense and host sociality: a comparative study of swallows and martins.
Sociality is associated with increased risks of parasitism, predation, and social competition, which may interact because social stress can reduce immunity, and parasitized individuals are more likely to fall prey to a predator. A mechanism allowing evolution of sociality in spite of high costs of parasitism is increased investment in antiparasite defenses. Here we show that the impact of parasites on host reproductive success was positively associated with the degree of sociality in the bird family Hirundinidae. However, the cost of parasitism in highly colonial species was countered by high levels of T- and B-cell immune responses. Investment in immune function among colonial species was particularly strong in nestlings, and among social species, this investment was associated with a relatively prolonged period of development, thereby leading to extended exposure to parasites. Thus, highly social species such as certain species of swallows and martins may cope with strong natural selection arising from parasites by heavy investment in immune function at the cost of a long exposure to nest parasites.


Does it exist a personality core of mental illness? A systematic review on core psychobiological personality traits in mental disorders.
Research investigating the relationship between mental disorders and personality traits leads to interesting results. Individuals affected by several mental disorders have been worldwide assessed according to the psychobiological model of personality. This review aims to explore which temperament and character traits are recurrent in mental disorders and to highlight what traits may be shared determinants or consequences of the expression of a mental disorder. Systematic search of Medline database between 1998 and 2011 has been conducted to select the studies exploring the Temperament and Character Inventory (TCI) dimensions in the most relevant axis I psychiatric disorders. Of the 110 studies that were retrieved, 88 met the inclusion/exclusion criteria and were analyzed. High HA (HA) and low self-directedness are recurrent and can be considered as a "personality core" regardless of the diagnosis. They may be risk factors and relapse-related, they can indicate incomplete remission or chronic course of mental disorders, and consistently influence patients' functioning. Furthermore, they can be modified by medications or psychotherapy and represent outcome predictors of treatments. This "core" may represent a personality diathesis to psychopathology. Relational environment can influence the development of both temperament and character, thus prevention of mental disorders should promote a positive development of these traits. Although further research is needed, psychotherapeutic interventions should be performed also considering that mental disorders could benefit from HA desensitization and SD reinforcement. Finally, these traits may be used to provide diagnostic, prognostic, quality of life and efficacy inferences on psychiatric treatments.


[Indications in sleep-apnea syndrome. When and why is further assessment meaningful?].
Forms of sleep apnea syndrome: Interrupted breathing and hypoventilation during sleep lead to sleep disorders and to cardiovascular sequelae. In the common obstructive sleep apnea syndrome (OSAS) apneas are related to intermittent obstruction of the upper airways. In the rarer central sleep apnea syndrome certain cardiovascular or central nervous system disorders lead to disturbed regulation of respiration connected with periodic breathing. Signs indicating OSAS: Loud, cyclic snoring, interrupted by cessation of breathing during sleep observed by relatives and excessive daytime to diurnal sleepiness indicate OSAS. Furthermore alteration of personality, headache in the morning, non-refreshing sleep and nocturnal choking sensations may indicate OSAS. When is evaluation necessary? Patients with complaints possibly induced by OSAS should be further evaluated since nocturnal application of continuous positive airway pressure (CPAP) by means of a nose mask and other treatment forms often lead to significant improvement of OSAS. In addition patients with untreated OSAS have an increased risk for car accidents and premature death as consequence of cardiovascular diseases. The type and extent of a supposed respiratory disorder is evaluated by means of a sleep study.


Childhood catatonia, autism and psychosis past and present: is there an 'iron triangle'?
To explore the possibility that autism, catatonia and psychoses in children are different manifestations of a single underlying form of brain pathology - a kind of 'Iron Triangle' of symptomatology - rather than three separate illnesses. Systematic evaluation of historical case literature on autism to determine if catatonic and psychotic symptoms accompanied the diagnosis, as is found in some challenging present-day cases. It is clear from the historical literature that by the 1920s all three diagnoses in the Iron Triangle - catatonia, autism and childhood schizophrenia - were being routinely applied to children and adolescents. Furthermore, it is apparent that children diagnosed with one of these conditions often qualified for the other two as well. Although conventional thinking today regards these diagnoses as separate entities, the presence of catatonia in a variety of conditions is being increasingly recognized, and there is also growing evidence of connections between childhood-onset psychoses and autism. Recognition of a mixed form of catatonia, autism and psychosis has important implications for both diagnosis and treatment. None of the separate diagnoses provides an accurate picture in these complex cases, and when given single diagnoses such as 'schizophrenia', the standard treatment options may prove markedly ineffective.


Association of Prostate Blood Flow with Male Lower Urinary Tract Symptoms.
To investigate the association of prostate blood flow (PBF) with lower urinary tract symptoms (LUTS) in aged males using Doppler spectral waveform (DSW) analysis. We performed a prospective analysis involving 133 aged males with clinical diagnosis of LUTS. DSW parameters (peak-systolic velocity (PSV), end-diastolic velocity (EDV), and resistive index (RI)) were measured at bilateral neurovascular bundles (NVB), periurethral, and capsular branches by Doppler transrectal ultrasound with the patient in the right lateral decubitus position. The associations of PBF parameters and the International Prostate Symptom Score (IPSS) were analyzed. Overall, total IPSS scores were significantly correlated with the RI of bilateral NVB vessels (r2 = 0.03, 0.04; p = 0.04, 0.02, respectively), and PSV of left NVB vessels. PSV of bilateral NVB vessels were associated with the storage score (p = 0.022 and p = 0.016), but not with the voiding score. The sum of the frequency and urgency score was also associated with EDV of both capsular and urethral branches (p = 0.043 and p = 0.009, respectively), and PSV of NVB vessels on both sides (p = 0.045 and p = 0.019, respectively). There is an association between PBF and LUTS, especially with storage symptoms. The findings may provide some insights in understanding the underlying pathophysiology of lower urinary tract dysfunction.


Long-term survival after hepatic cryosurgery versus surgical resection for metastatic colorectal carcinoma: a critical review of the literature.
To critically assess the evidence for long-term survival after hepatic resection and hepatic cryosurgery for metastatic colorectal cancer. The purpose of this review is to determine if a randomized controlled trial comparing these two treatment modalities is justified. A review of the medical literature from 1973 to 1995 using the MEDLINE and CANCERLIT databases. References were also retrieved from the bibliographies of identified articles and from experts in the field of hepatobiliary and pancreatic surgery. One hundred and seventy-eight studies were reviewed. Studies presenting original data on the results of hepatic resection or cryotherapy for colorectal liver metastases were selected. Studies were excluded if they did not present survival data longer than 2 years. Studies pertaining to resection for fewer than 60 patients with colorectal metastases to the liver were excluded. Data forms were designed before studies were examined in detail. All studies that met the inclusion and exclusion criteria were reviewed and the identified data extracted and tabulated. No controlled studies were identified, only case series. Four reports on hepatic cryosurgery and 9 on hepatic resection met the study criteria. The cryosurgery studies were methodologically poor; the resection studies were larger and more methodologically sound. The median follow-up for cryosurgery ranged from 12 to 28.8 months, that for resection 21 to 69 months. There is clear evidence that hepatic cryosurgery has a role in the management of selected patients with colorectal metastases to the liver. However, valid conclusions cannot be made about the 5-year survival rate. The results of the studies on hepatic resection in patients with colorectal metastases to the liver have greater validity and consistency, with 5-year survival rates of 20% to 40%. Although hepatic cryosurgery offers some unequivocal and other potential advantages over surgical resection for colorectal metastases to the liver, the published data do not support its use in patients with resectable disease outside a clinical trial, and do not yet justify a randomized trial. A study that collects prospective data on 2 groups of patients (resectable v. unresectable) who differ only in the anatomic location of their metastases within the liver is needed.


The Prognostic Impact of Hormonal Receptor and HER-2 Expression Discordance in Metastatic Breast Cancer Patients.
Hormone receptor (HR) and human epidermal growth factor receptor (HER2) discordance between primary and metastatic breast cancer lesions is common. However, its impact on long-term survival remains unclear. We aimed to determine the prognostic value of this discordance in patients with metastaticf breast cancer (MBC). A total of 270 patients with MBC who were underwent re-biopsy of progressive metastases at Zhejiang Cancer Hospital from January 1, 2012 to December 31, 2015 with patients consent and then review their primary tumors pathological findings. The HR and HER2 status in both primary and progressive metastatic lesions was determined by immunohistochemistry and/or fluorescence in situ hybridization. The discordance rates were correlated with the clinicopathologic characteristics, metastatic lesions, salvage treatment, and survival analysis in this population. A total of 142 (52.6%) MBC patients were diagnosed with discordant HR and HER2 status. Alterations in estrogen receptor (ER), progesterone receptor (PR), and HER2 status were observed in 20.70%, 37.78%, and 11.48% cases, respectively. Chemotherapy (P=0.0192) and endocrine therapy (P=0.048) significantly affected the conversion of HR status. Endocrine therapy was positively correlated with PR discordance (P=0.002), while ER discordance was associated with adjuvant chemotherapy (P=0.031). Survival analysis showed that ER status alterations between primary and metastatic lesions were associated with overall survival (P=0.002). The clinical prognosis was significantly worse with HR losses than with persistent HR positivity (P=0.023). In Cox multivariate analysis, the loss of HR expression and conversion to triple negative were independent prognostic indicators. Discordance in HR status between primary and metastatic lesions may impact the prognosis of MBC, and HR conversion has independent prognostic value.


From free pastures to penned in: The within-person effects of psychological reactance on side-hustlers' hostility and initiative in full-time work.
Multiple jobholding is increasingly common, particularly among full-time employees who have adopted side-hustles-income-generating work from the gig economy that is performed alongside full-time work. A distinguishing feature of side-hustles is substantial autonomy in the work's timing, location, and method. This autonomy has typically been portrayed as beneficial. We shift this consensus by developing a within-person model that suggests elevated side-hustle autonomy-relative to what is typical for that person-sets the employee on a course to feel "boxed in" by their full-time job. Drawing on psychological reactance theory, we argue that elevated autonomy in a side-hustle sensitizes employees to threats that restrict their control. As these employees shift to full-time work, we theorize that this sensitivity is associated with feelings of hostility that contribute to counterproductive behavior. We also propose, however, that side-hustle autonomy has benefits for full-time work-motivating employees to reassert control through increased initiative, thereby enhancing task performance. We explore the countervailing relationships between side-hustle autonomy and full-time work outcomes with a daily experience sampling study (ESM) of 101 full-time employees with side-hustles and their coworkers (Study 1) and a weekly ESM study of 100 full-time employees with side-hustles (Study 2). Taken together, we build and test theory about how employees' side-hustle autonomy exhibits within-person relationships that are a "mixed-bag" for their full-time work behavior. (PsycInfo Database Record (c) 2023 APA, all rights reserved).


Exendin-(9-39) Effects on Glucose and Insulin in Children With Congenital Hyperinsulinism During Fasting and During a Meal and a Protein Challenge.
The aim of this study was to assess whether exendin-(9-39) will increase fasting and postprandial plasma glucose and decrease the incidence of hypoglycemia in children with hyperinsulinism (HI). This was an open-label, four-period crossover study. In periods 1 and 2, the effect of three different dosing regimens of exendin-(9-39) (group 1, 0.28 mg/kg; group 2, 0.44 mg/kg; group 3, 0.6 mg/kg) versus vehicle on fasting glucose was assessed in 16 children with HI. In periods 3 and 4, a subset of eight subjects received either vehicle or exendin-(9-39) (0.6 mg/kg) during a mixed-meal tolerance test (MMTT) and an oral protein tolerance test (OPTT). Treatment group 2 showed 20% (P = 0.037) increase in the area under the curve (AUC) of fasting glucose. A significant increase in AUC of glucose was also observed during the MMTT and OPTT; treatment with exendin-(9-39) resulted in 28% (P ≤ 0.001) and 30% (P = 0.01) increase in AUC of glucose, respectively. Fasting AUC of insulin decreased by 57% (P = 0.009) in group 3. In contrast, AUC of insulin was unchanged during the MMTT and almost twofold higher (P = 0.004) during the OPTT with exendin-(9-39) treatment. In comparison with vehicle, infusion of exendin-(9-39) resulted in significant reduction in likelihood of hypoglycemia in group 2, by 76% (P = 0.009), and in group 3, by 84% (P = 0.014). Administration of exendin-(9-39) during the OPTT resulted in 82% (P = 0.007) reduction in the likelihood of hypoglycemia. These results support a therapeutic potential of exendin-(9-39) to prevent fasting and protein-induced hypoglycemia in children with HI.


The Concentration of Organic Acids in Cranberry Juice Modulates the Gut Microbiota in Mice.
A daily consumption of cranberry juice (CJ) is linked to many beneficial health effects due to its richness in polyphenols but could also awake some intestinal discomforts due to its organic acid content and possibly lead to intestinal inflammation. Additionally, the impact of such a juice on the gut microbiota is still unknown. Thus, this study aimed to determine the impacts of a daily consumption of CJ and its successive deacidification on the intestinal inflammation and on the gut microbiota in mice. Four deacidified CJs (DCJs) (deacidification rates of 0, 40, 60, and 80%) were produced by electrodialysis with bipolar membrane (EDBM) and administered to C57BL/6J mice for four weeks, while the diet (CHOW) and the water were ad libitum. Different parameters were measured to determine intestinal inflammation when the gut microbiota was profiled. Treatment with a 0% DCJ did not induce intestinal inflammation but increased the gut microbiota diversity and induced a modulation of its functions in comparison with control (water). The effect of the removal of the organic acid content of CJ on the decrease of intestinal inflammation could not be observed. However, deacidification by EDBM of CJ induced an additional increase, in comparison with a 0% DCJ, in the Lachnospiraceae family which have beneficial effects and functions associated with protection of the intestine: the lower the organic acid content, the more bacteria of the Lachnospiraceae family and functions having a positive impact on the gut microbiota.


[Nucleic acids of the lymphoid cells of the pregnant rat subjected to space flight and its progeny].
The space flight effect on the synthesis of nucleic acids in spleen lymphocytes in pregnant rats and their offspring was investigated. In addition to the inhibition of the DNA replicative function previously detected in males, activation of RNA synthesis was seen in the pregnant females. Such changes did not develop in the synchronous mockup controls. The 30-day and 100-day old pups of the rats flown in space during the last third of their pregnancy showed no changes in the DNA replicative function. The 100-day old animals displayed only a slight, when compared to the vivarium controls, decline of RNA synthesis which was produced by concomitant factors rather than by microgravity per se. It is supposed that microgravity-induced activation of RNA synthesis in spleen lymphocytes of pregnant rats is associated with the involvement of these cells in adaptive trophic processes that are to maintain plastic homeostasis of the fetus in an unusual environment.


[Properties of phosphodiesterase from Vipera lebetina venom immobilized on Agarose].
Preparations of phosphodiesterase from Vipera lebetina venom immobilized on agarose were obtained. The kinetic properties for the hydrolyses of various substrates of soluble and immobilized phosphodiesterase, e. g. effects of pH, temperature, substrate concentrations, etc., were compared. The values of Km, V and activation energy for the substrate hydrolyses were determined.


Organization, structure and data of the Hellenic Heart Registry on Percutaneous Coronary Interventions: a step forward towards outcomes research.
The Hellenic Heart Registry on Percutaneous Coronary Interventions (HHR-PCI) was a prospective, observational registry of patients with stable angina or acute coronary syndromes who underwent percutaneous coronary intervention (PCI) between January 2008 and October 2010. HHR-PCI was a database that used a secure web-based interface for data entry by individual users. All PCI centers and operators were invited to participate. The participating PCI centers were geographically divided into three main regions: Athens Metro Area, Thessaloniki Metro Area, and Rest of Greece. Indications, demographics, procedural characteristics, and in-hospital outcomes (death, myocardial infarction, cerebrovascular accidents) were recorded. Eighteen (18) centers participated in the registry (2008-2010) in a systematic fashion, entering complete data for 3441 patients (males 83.1%, mean age 64.1 years, 5521 lesions). PCI was elective in 47.1% of patients and was used to treat an acute coronary syndrome in 52.5%. There were 742 (21.6%) patients treated for ST-segment elevation myocardial infarction, with 225 (30.3%) receiving primary PCI (mean door-to-balloon time 140 minutes). The mean numbers of stents per lesion and per patient were 1.14 and 1.74, respectively, with drug eluting stents being used in 74.2% of coronary lesions. Periprocedural complications were observed in 105 patients (3.1%), while the incidence of in-hospital death, myocardial infarction, and cerebrovascular event were 0.5%, 1%, and 0.6%, respectively. HHR-PCI was the first web-based national PCI registry in Greece and has provided useful insights regarding the practice of interventional cardiology in this country. Efforts should be made to maintain and extend this type of PCI registry, with a view to improving quality and outcomes research in the field of interventional cardiology.


MetaNeuron: A Free Neuron Simulation Program for Teaching Cellular Neurophysiology.
MetaNeuron, a neuron simulation program, is an effective interactive tool for teaching cellular neurophysiology. The computer program simulates a wide range of neuronal behavior in its six lessons: i) Resting Membrane Potential, ii) Membrane Time Constant, iii) Membrane Length Constant, iv) Axon Action Potential, v) Axon Voltage Clamp, and vi) Synaptic Potential. The program is designed foremost as a platform for conducting neurophysiology experiments in silico. Neuronal parameters are easily modified and a virtual stimulator injects single or double current pulses into the neuron. Phenomena such as temporal summation of synaptic potentials, passive spread of a synaptic potential from the dendrite to the soma, the refractory period, families of voltage-clamp traces, and the reversal potential of synaptic responses, are easily illustrated in MetaNeuron. Responses are displayed graphically and can be measured with a cursor. Families of traces can be generated and viewed in rotatable 3D plots. Mac and Windows versions of the program can be downloaded, free of charge, onto individual student computers from the website www.MetaNeuron.org. A manual containing operating instructions, a description of the lessons, and exercises conducted on MetaNeuron, can also be downloaded for free.


An antagonist to vasoactive intestinal peptide affects cellular functions in the central nervous system.
A vasoactive intestinal peptide (VIP) antagonist was synthesized and used to investigate the interactions of VIP with its receptors present in the central nervous system (CNS). The VIP antagonist is a hybrid peptide consisting of a portion of VIP and a portion of neurotensin, designed to change the membrane permeability of the VIP portion. The hybrid antagonist displaced 80 to 90% of [125I]VIP binding to cell cultures from cerebral cortex, hippocampus or spinal cord. The displacement curve was biphasic, suggesting two binding sites. In the case of cortical astrocytes, the antagonist had a Ki of 45 pM at one site and a Ki of 74 nM at the other. At the lower affinity binding site, the antagonist was about 10-fold more potent than VIP in displacing radiolabeled VIP. The accumulation of cyclic AMP (cAMP) in VIP-stimulated cortical glia cultures was decreased by the new antagonist (EC50, 59 nM). This decrease in cAMP was greater than that achieved in the presence of other putative VIP antagonists. Finally, the addition of 1 nM hybrid antagonist to dissociated spinal cord cultures resulted in a 42% reduction in neuronal cell counts as compared with controls, and the EC50 of this effect was about 30 pM, which corresponded closely to the Ki of antagonist displacement of [125I]VIP binding at the high-affinity site. The antagonist appears to be a competitive blocker for both VIP-mediated increases in cAMP formation or VIP-associated maintenance of neuronal survival in spinal cord cultures. Thus, we describe a potent VIP antagonist which interacts with two functionally distinct VIP receptors in the CNS.


Canine peritoneal macrophages: cultivation and infection with Ehrlichia canis.
Canine peritoneal macrophages were obtained by repeated peritoneal lavage of dogs at 14 day intervals. Intraperitoneal administration of sterile mineral oil increased the leukocyte yield approximately 20-fold and the macrophage recovery approximately 35-fold. Cell recovery was maximum 7 to 21 days after administration of oil and declined slightly by 35 days. Restimulation with a 2nd injection of oil promptly revitalized cell recovery. Peritoneal macrophage cultures were well established by 6 days after seeding and were maintained for at least 30 days. Initially, cultured macrophages were mononuclear, but binuclear cells appeared after 6 days' cultivation and multinucleated giant cells were observed after 14 days. Canine peritoneal macrophages and peripheral blood monocytes were equally susceptible to infection with Ehrlichia canis. Infected cells were detected by 60 hours after inoculation, and replication was evident by 12 to 18 days.


Particle deposition in the trachea: in vivo and in hollow casts.
The pattern of deposition within the respiratory tract of potentially harmful particulates is a major factor in assessing any risk from individual and community exposures. Although the trachea is the most easily observed of the conductive airways, very little information concerning its particle collection characteristics is available, information which is essential for a complete and realistic description of particle deposition patterns within the entire respiratory tract. Data on tracheal deposition are also needed for development of accurate predictive models for particle deposition. The pattern of particle deposition in the trachea, and its relation to air flow, was studied in a hollow cast of the human larynx-tracheobronchial tree. Results were compared with data obtained in humans in vivo and from previous studies in hollow casts. In addition, the relevance of tracheal deposition in the hollow cast test system to deposition in vivo was examined by a direct comparison of deposition in a cast prepared from the lungs of donkeys previously studied in a series of in vivo tests. The disturbance of the air flow within the trachea caused by the larynx promoted the deposition of suspended particulates throughout the length of the trachea, and especially in proximal regions. This proximal deposition was due both to direct impaction from the air jet coming from the glottis and to effects of the tubulent flow. Turbulence produced inhomogenous deposition patterns within the trachea for particles of all sizes, although its effect was more pronounced as size decreased. Tracheal deposition in the human cast was within the range of normal in vivo tracheal depostion only when a larynx was used during cast test exposures; this emphasizes the need for the use of realistic experimental test systems for the study of particle deposition patterns. The relative patterns of deposition in casts of the donkey trachea and in the same tracheas in vivo were similar.


Clinical characteristics, timing of peak responses and safety aspects of two dosing regimens of the glucagon stimulation test in evaluating growth hormone and cortisol secretion in adults.
Weight-based (WB: 0.03 mg/kg) and fixed dose (FD: 1-1.5 mg) regimens of the glucagon stimulation test (GST) have been used to evaluate GH and cortisol secretion in children and adults, respectively. However, experience of the WB regimen in assessing GH and cortisol secretion in adults are limited. We describe a multicenter experience using WB and FD regimens in evaluating GH and cortisol secretion in adults suspected of GH deficiency and central adrenal insufficiency. Retrospective case series of GSTs (n = 515) performed at five tertiary centers. Peak and nadir glucose, and peak GH and peak cortisol responses occurred later with WB (mean dose: 2.77 mg) compared to FD (mean dose: 1.20 mg) regimens. Main side-effects were nausea and vomiting, particularly in younger females. Nausea was comparable but vomiting was more frequent in the WB regimen (WB: 10.0 % vs FD: 2.4 %; P < 0.05). Peak and nadir glucose, ΔGH, and peak and Δcortisol were higher in the WB regimen. In both regimens, age correlated negatively with peak cortisol levels, and body mass index (BMI), fasting, peak and nadir glucose correlated negatively with peak GH levels. WB and FD regimens can induce adult GH and cortisol secretion, but peak responses occur later in the WB regimen. Both regimens are relatively safe, and vomiting was more prevalent in the WB regimen. As age, BMI, and glucose tolerance negatively correlated with peak GH and cortisol levels, the WB regimen may be more effective than the FD regimen in older overweight glucose intolerant patients.


Simple and Fast DNA Based Sensor System for Screening of Small-Molecule Compounds Targeting Eukaryotic Topoisomerase 1.
Background: Eukaryotic topoisomerase 1 is a potential target of anti-parasitic and anti-cancer drugs. Parasites require topoisomerase 1 activity for survival and, consequently, compounds that inhibit topoisomerase 1 activity may be of interest. All effective topoisomerase 1 drugs with anti-cancer activity act by inhibiting the ligation reaction of the enzyme. Screening for topoisomerase 1 targeting drugs, therefore, should involve the possibility of dissecting which step of topoisomerase 1 activity is affected. Methods: Here we present a novel DNA-based assay that allows for screening of the effect of small-molecule compounds targeting the binding/cleavage or the ligation steps of topoisomerase 1 catalysis. This novel assay is based on the detection of a rolling circle amplification product generated from a DNA circle resulting from topoisomerase 1 activity. Results: We show that the binding/cleavage and ligation reactions of topoisomerase 1 can be investigated separately in the presented assay termed REEAD (C|L) and demonstrate that the assay can be used to investigate, which of the individual steps of topoisomerase 1 catalysis are affected by small-molecule compounds. The assay is gel-free and the results can be detected by a simple colorimetric readout method using silver-on-gold precipitation rendering large equipment unnecessary. Conclusion: REEAD (C|L) allows for easy and quantitative investigations of topoisomerase 1 targeting compounds and can be performed in non-specialized laboratories.