Evidence for altered circular smooth muscle cell function in lower esophageal sphincter of W/Wv mutant mice.
Nitrergic neurotransmission to gut smooth muscle is impaired in W/W(v) mutant mice, which lack intramuscular interstitial cells of Cajal (ICC-IM). In addition, these mice have been reported to have smaller amplitude unitary potentials (UPs) and a more negative resting membrane potential (RMP) than control mice. These abnormalities have been attributed to absence of ICC-IM, but it remains possible that they are due to alterations at the level of the smooth muscle itself. Amphotericin-B-perforated patch-clamp recordings and Ca(2+) imaging (fura 2) were compared between freshly isolated single circular smooth muscle cells (CSM) from W/W(v) mutant and control mice lower esophageal sphincter (LES). There was no significant difference in seal resistance, capacitance, or input resistance in response to applied electrotonic current pulses between CSM cells from W/W(v) mutants and controls. Compared with control mice, RMP was more negative and UPs significantly smaller in CSM cells from mutant mice LES. Administration of caffeine induced an inward current in cells from both mutant and control mice, but the current density was significantly larger in cells from W/W(v) mutants. Membrane potential hyperpolarization induced by sodium nitroprusside was larger in cells from control mice vs. W/W(v) mutants. In addition, intracellular Ca(2+) transients induced by caffeine were significantly increased in cells from mutants. These findings indicate that LES CSM is abnormal in W/W(v) mutant mice. Thus some physiological functions attributed to ICC-IM based on experiments in smooth muscle of ICC deficient mice may need to be reconsidered.


Hypoxia and vitamin D differently contribute to leptin and dickkopf-related protein 2 production in human osteoarthritic subchondral bone osteoblasts.
Bone remodelling and increased subchondral densification are important in osteoarthritis (OA). Modifications of bone vascularisation parameters, which lead to ischemic episodes associated with hypoxic conditions, have been suspected in OA. Among several factors potentially involved, leptin and dickkopf-related protein 2 (DKK2) are good candidates since they are up-regulated in OA osteoblasts (Obs). Therefore, in the present study, we investigated the hypothesis that hypoxia may drive the expression of leptin and DKK2 in OA Obs. Obs from the sclerotic portion of OA tibial plateaus were cultured either under 20% or 2% oxygen tension in the presence or not of 50 nM of 1,25-dihydroxyvitamin D3 (VitD3). The expression of leptin, osteocalcin, DKK2, hypoxia-inducible factors (Hif)-1α and -2α was measured by real-time polymerase chain reaction and leptin production by enzyme-linked immunosorbent assay (ELISA). The expression of Hif-1α, Hif-2α, leptin and DKK2 was reduced using silencing (si) RNA technique. Signalling pathway of hypoxia-induced leptin was investigated by western blotting and mitogen-activated protein kinase (MAPK) inhibitors. As expected, hypoxia stimulated the expression of Hif-1 and Hif-2. The expression of leptin and DKK2 in Obs was also stimulated 7-fold and 1.8-fold respectively (p<0.05) under hypoxia. Interestingly, whereas VitD3 stimulated leptin and DKK2 expression 2- and 4.2-fold under normoxia, it further stimulated it to 28- and 6.2-fold under hypoxia (p<0.05). The hypoxia-induced leptin production was confirmed by ELISA, particularly in presence of VitD3 (p<0.02). Compared to Obs incubated in the presence of siScramble RNAs, siHif-2α inhibited VitD3-stimulated leptin mRNA and protein levels by 70% (p=0.004) and 60% (p<0.02), respectively while it failed to significantly alter the expression of DKK2. SiHif-1α has no effect on these genes. Immunoblotting showed that VitD3 greatly stabilized Hif-2α under hypoxic condition. The increase in leptin expression under hypoxia was also regulated, in addition to the role of Hif-2α, by p38 MAPK (p<0.03) and PI 3-kinase (p<0.05). Finally, we demonstrated that the expression of leptin and DKK2 were not related to each other under hypoxia. Hypoxic conditions via Hif-2 regulation trigger Obs to produce leptin particularly under VitD3 stimulation whereas DKK2 is mainly regulated by VitD3 rather than hypoxia.


[The spatial organization of the vestibulospinal neurons in the guinea pig].
Quantitative investigation of the spatial organization of vestibulospinal neurons has been performed on a guinea pig using the horseradish peroxidase retrograde transport method. After unilateral injection in the upper cervical and low thoracic spinal cord the labelled neurons were found unilaterally in the lateral vestibular nucleus and bilaterally in descending and medial vestibular nuclei. Distribution of vestibulospinal neurons within the brainstem has been analyzed. It was revealed that projections of some neurons of medial and descending vestibular nuclei coincided with spinal projections of cortical and rubral descending pathways. Functional significance of the vestibulospinal system in the supraspinal motor control was discussed.


Reinterpreting the vibrational structure in the electronic spectrum of the propargyl cation (H2C3H+) using an efficient and accurate quantum model.
The B̃1A1 ← X̃1A1 absorption spectra of propargyl cations H2C3H+ and D2C3D+ were simulated by an efficient two-dimensional (2D) quantum model, which includes the C-C stretch (v5) and the C≡C stretch (v3) vibrational modes. The choice of two modes was based on a scheme that can identify the active modes quantitively by examining the normal coordinate displacements (∆Q) directly based on the ab initio equilibrium geometries and frequencies of the X̃1A1 and B̃1A1 states of H2C3H+. The spectrum calculated by the 2D model was found to be very close to those calculated by all the higher three-dimensional (3D) quantum models (including v5, v3, and another one in 12 modes of H2C3H+), which validates the 2D model. The calculated B̃1A1 ← X̃1A1 absorption spectra of both H2C3H+ and D2C3D+ are in fairly good agreement with experimental results.


[Physicians and the mass media. Opinion of physicians on the coverage of medicine, health policy and the medical profession by the mass media].
Every fourth physician disapproves of mass media coverage of health policy questions. Four out of ten physicians are dissatisfied with the way scientific medical issues are covered, and seven out of ten are dissatisfied with articles and programmes about the medical profession. The physicians' dissatisfaction was predicted by a low level of perceived job autonomy and a high level of perceived unrealistic expectations from patients, families, superiors and politicians. The doctors' disapproval of press coverage of the medical profession increased with perceived stress. The dissatisfaction was clearly greater among younger than among older physicians. Internists and surgeons tended to be more dissatisfied than physicians in other specialties. Dissatisfaction with mass media was not, however, a question of personal grudge: respondents who felt that they themselves had been unfairly spoken of by the media did not disapprove of the media coverage of health issue, or of the medical profession in general, to any greater degree than did doctors who had no such personal complaints.


[The perception by the population of the functions and components of the primary care team].
To find out how the population in Health Area IV of Navarra (Tafalla) see the new components of the primary care team (PCT) and their health education functions. A crossover descriptive study. SITE. Health Area IV of Navarra (Tafalla). 816 people aged 18 or over and who lived in the area under study made up a sample stratified according to age, gender and where they lived. Interviewees were selected through the random routes method. A personal interview with a questionnaire was carried out in the home of the participants during March and April, 1988. There was widespread ignorance of what the PCT meant; paradoxically, this ignorance was significantly higher (p = 0.014) in the base areas where EAP had been set up (80%) than in those areas where it had not (73%). In spite of this, in the Tafalla base area the components of the PCT had begun to be understood. Only 7% replied that they knew other functions, apart from health care, of the PCTs: here, it was in the areas where PCTs had been set up that there was significantly greater knowledge (p = 0.0004). A large percentage of the population is ignorant regarding new elements (staff and functions) introduced into the PCT.


Analysis of Epigenetic Changes in Vitamin D Pathway Genes in Rheumatoid Arthritis Patients.
Rheumatoid arthritis (RA) is an autoimmune inflammatory disease with complex etiopathogenesis launched by multiple risk factors, including epigenetic alterations. RA is possibly linked to vitamin D that is epigenetically active and may alter DNA methylation of certain genes. Therefore, the study aimed to evaluate the relationship between DNA methylation status of vitamin D signaling pathway genes (VDR, CYP24A1, CYP2R1), vitamin D level and associations with RA. Totally 76 participants (35 RA patients and 41 healthy controls) were enrolled from a case-control vitamin D and VDR gene polymorphisms study regarding age and vitamin D concentration. CpG islands in promoter regions of the VDR, CYP24A1, CYP2R1 genes were chosen for DNA methylation analysis by means of pyrosequencing. Chemiluminescent microplate immunoassay was used to assess 25(OH)D serum levels. RA clinical data, i.e. the disease activity score C-reactive protein 28 (DAS28 - CRP) as well as patient-reported outcome questionnaires were recorded. The study showed similar methylation pattern in the promoter regions of vitamin D pathway genes in RA and control group with p>0.05 (VDR gene 2.39% vs. 2.48%, CYP24A1 gene 16.02% vs. 15.17% and CYP2R1 2.53% vs. 2.41%). CYP24A1 methylation intensity was significantly higher in compare to methylation intensity of VDR and CYP2R1 genes in both groups (p<0.0001). A tendency of higher vitamin D concentration in cases having methylated VDR (57.57±28.93 vs. 47.40±29.88 nmol/l), CYP24A1 (53.23±26.22 vs. 48.23±34.41 nmol/l) and CYP2R1 (60.41±30.73 vs. 44.54±27.63 nmol/l) genes and a positive correlation between VDR, CYP2R1 methylation intensity and vitamin D level in RA affected participants was revealed (p>0.05). A significantly higher CYP24A1 methylation intensity (p=0.0104) was detected in blood cells of vitamin D deficient (<50 nmol/l) RA patients vs. vitamin D deficient controls. Our data suggests some indirect associations between DNA methylation status of vitamin D pathway genes and vitamin D level in RA.


Immunosuppression associated with SJL/J murine lymphoma. I. Suppression of cell-mediated immune responses after tumor transplantation.
The proliferative responses of peripheral blood lymphocytes of inbred SJL/J mice to the mitogens phytohemagglutinin and concanavalin A were evaluated serially in individual animals before and after induction of lymphomas by transplantation. Tumor-bearing mice exhibited marked suppression of mitogen responsiveness. Proliferative responses in mixed lymphocyte-tumor cell culture and mixed lymphocyte culture were also suppressed. Suppression of responses was tumor related, and responsiveness was restored in animals whose tumors regressed. There was no deficiency of responder T-cells in the peripheral blood of tumor-bearing animals, and suppressor cell activity was not demonstrable in a number of in vitro assays. However, plasma from tumor-bearing animals exhibited potent immunosuppressive activity. The presence of plasma suppressive activity was similarly tumor related. Thus this study demonstrates nonspecific suppression of cell-mediated immune responses following tumor induction, apparently mediated by a plasma suppressor factor.


Brain metastases from epithelial ovarian cancer--report of cases.
Epithelial ovarian cancer is one of the gynecological malignancies most commonly diagnosed late and one of the principal causes of mortality among women. The majority of women present with advanced disease. However, 5-year survival of patients with ovarian cancer has improved in recent years. Brain metastases from epithelial ovarian cancer are rare but in the last few years the incidence of brain complications seems to be increasing. Among all patients registered as having epithelial ovarian cancer at the Department of Oncology, Division of Gynecological Oncology, Poznan University of Medical Sciences, Poland between August 1998 and March 2008, four patients (4/669) who developed central nervous system (CNS) metastases were identified. Patients with symptoms of the CNS were evaluated by a neurologist, with a CT scan of the brain. The most common symptom of brain metastases are headaches which occur in 40-50% of patients. Because of the rarity of these patients, the optimal treatment for brain metastases is ill-defined. Brain metastasis usually appears with a poor prognosis, however early diagnosis and aggressive multimodal treatment can improve the quality of life in patients.


Effect of macrolide antibiotics on nitric oxide synthase and xanthine oxidase activities, and malondialdehyde level in erythrocyte of the guinea pigs with experimental otitis media with effusion.
Although the long-term administrations of macrolide antibiotics are effective for diffuse panbronchiolitis, otitis media with effusion (OME), and some other diseases, their mechanism of action has not been fully understood. In order to elucidate the mechanisms of possible effects of macrolide antibiotics on activities of erythrocyte nitric oxide synthase (NOS), xanthine oxidase (XO), and malondialdehyde (MDA) levels in experimental OME, we aimed to evaluate the effect of macrolide antibiotics (erythromycin, azithromycin, roxithromycin, and clarithromycin) using an experimental guinea pig otitis media model. Erythrocyte NOS, XO activities, and MDA level were measured in all groups. Erythrocyte NOS activities were significantly higher in erythromycin-, azithromycin-, roxithromycin-, and clarithromycin-treated groups than in the experimental group. Erythrocyte XO activities were significantly lower in erythromycin-, azithromycin-, roxithromycin-, and clarithromycin-treated groups than in the control group. However, erythrocyte XO activities in experimental group were significantly higher than those of control group. Erythrocyte MDA levels were significantly lower in erythromycin-, azithromycin-, roxithromycin-, and clarithromycin-treated groups than those of the experimental group. The MDA levels in erythromycin- and roxithromycin-treated groups were significantly higher than those of azithromycin-treated group. The MDA levels in azithromycin-treated group were significantly lower than those of roxithromycin-treated group. In conclusion, the present study shows that the macrolide antibiotics (erythromycin, azithromycin, roxithromycin, and clarithromycin) increase NOS activity, decrease XO activity and MDA level, which is an important indicator of oxidative stress.


An observational study of dengue fever in a tertiary care hospital of eastern India.
Dengue fever (DF) has become a significant resurgent tropical disease in the past 20 years all over the globe. The recent outbreak in West Bengal has once again underlined our failure in vector control and prevention. Our study outlines the clinical spectrum as well as the geographical expansion of the disease beyond urban confines. All patients with acute febrile illness positive for IgM antibody for Dengue virus were taken as cases. The patients were subjected to clinical examination and baseline investigations so as to fill in a structured proforma. The total number of patients were 180 of whom 92 (51.1%) were male and 88 (48.9%) were female. The maximum number of patients belonged to the age group 20-29 years (26.9%). The patients mostly hailed from Kolkata followed by Nadia, 24- Parganas(S), Murshidabad and Midnapur. The most common presentation apart from fever and bodyache were gastrointestinal symptoms. 42% patients complained of abdominal pain, 24% had vomiting, 9.6% diarrhoea. Bleeding manifestations occurred in 23% of patients. CNS features were documented 10.4%. Case fatality came out to be 3.8%. Investigations revealed thrombocytopenia in 55% and leucopenia 32.7%, transaminitis in 72% Evidence of organomegaly (22.2%) and serositis (42%) were detected. Complications included intracranial haemorrhage, DIC, pancreatitis, myocarditis and even a solitary case of splenic rupture. The current outbreak was affecting both the genders equitably and mostly the younger age group from rural as well as urban areas. A febrile illness characterised by myalgia, mild bleeding and gastrointestinal symptoms, it was more or less promptly responsive to early conservative therapy like fluids, FFP and platelet transfusion where required.


Electroencephalographic Correlates of Suicidal Ideation in the Theta Band.
Previous studies have indicated that the abnormalities of brain activity are associated with suicidal behavior. However, few studies have explored neurophysiologic biomarkers of suicidal ideation (SI) in healthy subjects. Here, we studied alterations of theta quantitative EEG (QEEG) in frontal regions associated with SI in young healthy subjects. Resting QEEG data were recorded from 90 young healthy subjects. The subjects were divided into two group based on Scale for Suicidal Ideation (SSI) scores: a high SSI group (n = 33) and a low SSI group (n = 57). Individual frontal electrodes (Fp1, Fp2, F7, F3, Fz, F4, F8) and central midline electrodes (FCz, Cz) were examined for absolute and relative power in the theta band. Clinical state and social support was assessed using the Hospital Anxiety and Depression Scale and Functional Social Support Questionnaire. We found that theta absolute and relative power in all channels was positively correlated with SSI, except Fp1, F7 and F8. The high SSI group showed higher theta relative power at F3, Fz, FCz, and Cz than the low SSI group. Theta relative power in the fronto-central region was significantly increased in the high SSI group compared to that in the low SSI group. Hierarchical regression analysis showed that SSI score was the most powerful predictor of fronto-central theta power. The findings suggest that brain electrical activity at the fronto-central region may be associated with differences in SI in young healthy subjects. EEG activity in theta band has clinical potential as a biomarker for preventing suicide.


Investigations of the Equilibrium Conditions in Plumes of Laser Desorbed Sinapic Acid with Amino Acid Analytes: Influence of Sample Preparation and Matrix to Analyte Ratio.
The equilibrium nature of a plume of laser desorbed material is examined through the application of a previously developed thermodynamic model to the ion signals observed in 337 nm MALDI mass spectra of mixtures of the matrix sinapic acid with the amino acids alanine, valine, isoleucine, and phenylalanine. Samples are prepared using both conventional dried-droplet and solvent-free methods for comparison. The relative yield of protonated amino acid is shown to increase as the amino acid gas-phase basicity increases for both sample preparation methods. Matrix gas phase basicity values extracted from the equilibrium plots are shown to be in good agreement ([M - H+]• 876 kJ/mol and [M] 879 kJ/mol) with published experimental values supporting a mechanism wherein the protonated sinapic acid and/or the matrix radical cation act as the proton donor species. These experiments further reveal that there is a large difference in the extracted plume effective temperatures with the solvent-free method yielding lower effective temperatures as compared to the dried-droplet sample preparation, e.g., 552 K versus 1296 K, respectively, at M/A 1:1 (mole/mole). In addition, these experiments suggest that plume effective temperatures decrease as the relative amount of matrix deposited with the analyte increases, regardless of the sample preparation method. Cumulatively, these observations suggest that the crystalline solid allows more efficient transfer of the photoexcitation energy during the sample desorption step, as compared to the solvent-free sample, and/or collisional cooling is more effective for the plume of material desorbed from the solvent-free sample as compared to the conventional dried-droplet sample.


[Hemispheric differences of simple reaction times in non-psychotic unipolar depressed patients].
Simple reaction times to visual stimuli were studied in 8 patients affected by depression and in 8 normal subjects. Regardless of the hand used, the condition of depression produced a specific lengthening of the responses to stimuli located in the left visual field. It is concluded that the right hemisphere plays a major role in modulation of negative emotional behaviour.


Palbociclib:CDK4/6 inhibition in the treatment of ER-positive breast cancer.
Maintaining cell-cycle control has become a mainstay in treatment for many cancers. Cell-cycle manipulation can be especially valuable in breast cancer tumor cells that will often express hormone receptors that are amenable to anti-hormone receptor-targeted therapies. Despite these treatments, patients often progress, leading to other targeted agents being investigated to help promote progression-free survival. Cyclin-dependent kinases (CDKs) have been identified as contributors in the process of cell division. Combining inhibitors of CDKs with traditional endocrine treatments has shown significant progression-free survival in patients with metastatic breast cancer. One such CDK inhibitor, palbociclib, has shown great promise in the treatment of hormone receptor-positive breast cancer. In this article we review the traditional hormonal treatments of breast cancer, how CDK inhibition is beneficial in the treatment of this disease, and the preclinical and clinical data supporting the use of this medication.


The potential for microchip-automated technology to improve enrichment practices.
The greatest limitation involved with providing enrichment and evaluating its effects on captive mammals is a lack of available staff time [Hoy et al., 2009]. One method to overcome this is through the use of automated husbandry systems to dispense enrichment to individual animals by using implantable radio frequency microchips. An international multi-institutional survey was conducted to determine whether the personnel involved with management, research, and husbandry of captive mammals would support the development of such automated husbandry systems. The survey was also designed to establish which applications of the system would be considered to be most beneficial. Overall, there was a very high level of interest by survey respondents in adopting automated husbandry systems and all potential applications were considered beneficial to varying degrees. These results, together with the relatively low cost of installation and operation, demonstrate that in the future automated enrichment could be an important and effective means of enhancing captive animal management.


[Abundant erythroblastic islands in bone marrow aspirate in a case of polycythemia vera. Various considerations on this anatomo-functional structure].
Despite the initial description, more than 30 years ago, of an erythroblastic island (EI), little is known about the mechanisms of union and interaction between the different cells of this anatomical and functional bone marrow structure. The observation of entire Els is extremely uncommon in normal bone marrow aspirates because of their dispersion during spread. A case report is described in which a 73-year-old woman, diagnosed 20 years ago with polycythaemia vera, showed multiple Els in a bone marrow aspirate. Their immunophenotyping revealed deficiency of several integrins. The ultrastructural study indicated a marked erythroblastic rophalocytosis. A hypothesis is suggested in view of these observations.


Single-shot network analyzer for extremely fast measurements.
A new instrument for fast measurement of frequency response of high-bandwidth optical and electronic devices is reported. Single-shot frequency spectrum measurements are enabled by time-stretch technology. An extremely fast measurement time of 27 ns is reported for the instrument. The reported instrument enables single-shot impulse response measurements with a 40 GHz bandwidth, which could be extended to beyond 100 GHz by using a faster electro-optic modulator. An ultra-low jitter of 20.5 fs is reported for the proposed instrument. The impulse responses measured using this technique are shown to correspond consistently with the manufacturer's specifications for the device under test. The reported instrument makes possible high-speed network parameter measurements, thereby enabling high-speed production-level testing of high-bandwidth opto-electronic devices/circuits/subsystems/systems and complex permittivity measurement of dielectric materials at a much reduced test time, lowering the test costs in a production environment.


Residual ethylene oxide in hollow-fiber dialyzers.
The deaeration and extraction kinetics of ethylene oxide (ETO) in hollow-fiber dialyzers were examined. The investigations showed that both deaeration and extraction can be described by two additive first-order kinetic expressions. The ETO content of membranes decreased rapidly at first, followed by a second slower decrease. Even after prolonged storage time, residual ETO was still detectable in the milligram range. These residuals cannot be eliminated by the usual rinsing of the dialyzer prior to use.


Serum albumin-lipid membrane interaction influencing the uptake of porphyrins.
It is frequently observed in pharmaceutical practice that entrapped substances are lost rapidly when liposomes are used as carriers to introduce substances into cells. The reason for the loss is the interaction of serum components with liposomes. To elucidate the mechanism of this phenomenon the partition of mesoporphyrin (MP) was systematically studied in model systems composed of various lipids and human serum albumin (HSA). As surface charge is an important factor in the interaction, neutral (1, 2-dimyristoyl-sn-glycero-3-phosphatidylcoline, DMPC) and negatively charged (1,2-dimyristoyl-sn-glycero-3-phosphatidylcoline/1, 2-dimyristoyl-sn-glycero-3-phosphatidylglycerol, DMPC/DMPG = 19/1 w/w) lipids were compared. The liposome/apomyoglobin system was the negative control. The size distribution of sonicated samples was carefully analyzed by dynamic light scattering. Constants of association of MP to the proteins and to the liposomes were determined: K(p,1) = (2.5 +/- 0.7) x 10(7) M(-1), K(p,2) = (1.0 +/- 0.7) x 10(8) M(-1), K(L,1) = (1.3 +/- 0.3) x 10(5) M(-1), and K(L,2) = (3.2 +/- 0.6) x 10(4) M(-1) for HSA, apomyoglobin, DMPC, and DMPC/DMPG liposomes, respectively. These data were used to evaluate the partition experiments. The transfer of MP from the liposomes to the proteins was followed by fluorescence spectroscopy. In the case of apomyoglobin, the experimental points could be interpreted by ruling out the protein-liposome interaction. In the case of HSA, the efflux of MP from the liposomes was strongly inhibited above a critical HSA concentration range for negatively charged vesicles. This effect was interpreted as the result of HSA coat formation on the liposome surface. This direct interaction is significant for small liposomes. The interpretation is fully supported by differential scanning calorimetry experiments.


Costs of illness: United States, 1980.
The total costs of illness and injury in the U.S. civilian noninstitutionalized population in 1980 amounted to $381.7 billion. The direct costs of illness and injury--resource expenditures for the diagnosis, treatment, and management of medical and dental conditions--were $153.9 billion, or 40.3 percent of total costs. Indirect costs--economic losses from morbidity and mortality--were $227.9 billion, or 59.7 percent of total costs. Of indirect costs, $104.9 billion resulted from productivity losses because of morbidity, and $123.0 billion represent the present value of lost productivity from premature mortality based on a net effective discount rate of 4 percent. These estimates, based on data from the 1980. National Medical Care Utilization and Expenditure Survey (NMCUES), differ from other estimates of the costs of illness and injury in 1980 (Gibson and Waldo, 1982; Rice, Hodgson, and Kopstein, 1985). The differences, which can be resolved, are attributable to two major factors: (1) NMCUES includes only the civilian noninstitutionalized population, but the other estimates include the institutionalized population and the military; and (2) NMCUES indirect cost estimates for the population unable to work include persons who were retired for health reasons in 1979 and 1980, disabled homemakers, and other persons who were disabled for the entire year 1980 but were not retired for health reasons in 1979, but the Rice et al. estimates do not include the last two categories in the population unable to work. The principal NMCUES findings on the total costs of illness in the civilian noninstitutionalized population reinforce the importance of considering distributional effects. Persons 65 years of age and over represent one-tenth of this population yet account for more than one-fourth of direct costs and more than their share of total costs, even though the institutionalized elderly are excluded. More than two-thirds of total costs for this age category are accounted for by direct costs. Direct costs also account for more than two-thirds of total costs for people under 17 years of age. However, this youngest age category, which constitutes over one-fourth of the civilian noninstitutionalized population, generates only 12.3 percent of direct costs. In contrast, indirect costs account for well more than 60 percent of total costs for the working-age population (17-64 years of age). Within the working-age population, per capita direct costs are highest among persons who are not full participants in the work force, many of whom are not working full time or at all because of injury or ill health.(ABSTRACT TRUNCATED AT 400 WORDS)


The neutrophil function of uremic patients treated by hemodialysis or CAPD.
The phagocytic, bactericidal and chemotactic function of blood neutrophils in uremic patients treated either by CAPD or hemodialysis was assessed in vitro. The phagocytosis was normal in 11 patients on CAPD treatment while it was significantly impaired in 13 patients on maintenance hemodialysis (HD) treatment. In addition the chemotactic function of neutrophils seemed to be better on CAPD treatment, when heat-inactivated serum, zymosan or casein were used as chemoattractants, but the difference was significant only with heat-inactivated serum. In contrast the chemotactic response to Staphylococcus aureus was better in patients on HD treatment. The intracellular killing of bacteria was normal in both patient groups. The more stable metabolic control of acid-base and electrolyte balance and the better clearance of middle molecules could explain the better performance of uremic neutrophils in the patients on CAPD treatment.


Molecular Analysis of the Contribution of Alkaline Protease A and Elastase B to the Virulence of Pseudomonas aeruginosa Bloodstream Infections.
Pseudomonas aeruginosa is a major cause of nosocomial bloodstream infections. This microorganism secretes two major proteases, alkaline protease A (AprA) and elastase B (LasB). Despite several in vitro studies having demonstrated that both purified proteases cleave a number of components of the immune system, their contribution to P. aeruginosa bloodstream infections in vivo remains poorly investigated. In this study, we used a set of isogenic mutants deficient in AprA, LasB or both to demonstrate that these exoproteases are sufficient to cleave the complement component C3, either soluble or deposited on the bacteria. Nonetheless, exoprotease-deficient mutants were as virulent as the wild-type strain in a murine model of systemic infection, in Caenorhabditis elegans and in Galleria mellonella. Consistently, the effect of the exoproteases on the opsonization of P. aeruginosa by C3 became evident four hours after the initial interaction of the complement with the microorganism and was not crucial to survival in blood. These results indicate that exoproteases AprA and LasB, although conferring the capacity to cleave C3, are not essential for the virulence of P. aeruginosa bloodstream infections.


The fetal and maternal consequences of gestational diabetes mellitus.
Gestational diabetes mellitus (GDM) is a type of diabetes that presents during pregnancy and usually disappears shortly after a woman gives birth. Better recognition of the risk factors of GDM, combined with more universal screening for the disease in many countries, has led to the increased detection of GDM along with other forms of pregestational diabetes. There is growing evidence that GDM significantly increases the risk of a number of short- and long-term adverse consequences for the fetus and mother, the most significant of which is a predisposition to the development of metabolic syndrome and Type 2 diabetes. Maternal and childhood obesity as well as cardiovascular disease are also potential long-term consequences of GDM. On the other hand, there is a growing body of evidence suggesting that the risk of many of these consequences can be significantly reduced or eliminated by aggressive treatment of GDM. There remains, however, a great deal of controversy over when to begin screening for hyperglycemia in pregnancy and at what level of hyperglycemia should aggressive intervention be initiated.


Total RNA Isolation from Separately Established Monolayer and Hydrogel Cultures of Human Glioblastoma Cell Line.
Astrocytoma is an invasive carcinoma occurring in the nervous system and currently lacks effective treatment options. A deeper understanding of the mechanisms of tumorigenesis and tumor progression is needed in order to develop novel therapeutic strategies. Recent advances in in vitro culture systems have demonstrated that the use of three-dimensional (3D) culture models could be more relevant for this purpose as compared to monolayer or two-dimensional (2D) models due to their resemblance to in vivo cancer pathology. High-throughput techniques such as RNA sequencing, microarray analyses and cloning could provide useful insights into the relevance of these systems to the native tissue. Previous studies have reported RNA extraction protocols needed for such applications. We have modified these protocols to suit the isolation of total RNA from monolayer and hydrogel cultures of astrocytoma established using basement membrane matrix, Geltrex™. We have used this method to demonstrate the differences in the expression of genes involved in autophagy, a process deregulated in many cancer types, in monolayer and hydrogel cultures using quantitative polymerase chain reaction (qPCR). This protocol can be adopted by the researchers who wish to understand the molecular basis of gene expression in hydrogel cultures of normal as well as cancer cell lines.


[Current status of nuclear medicine. Clinical application of FDG-PET for cancer diagnosis. Malignant lymphoma].
Malignant lymphoma is one of the tumors that show high FDG uptake, enabling the accurate detection of small involved lesions by FDG-PET. It is useful in both initial staging and follow-up study. In initial staging, FDG-PET is superior to CT from the aspects of sensitivity and specificity, and it is reported that the therapeutic protocol has been changed in 21% of patients. In the follow-up of therapy and detection of recurrence, the accuracy of residual disease is reported to be more than 90%. Among the different pathological types of lymphoma, Hodgkin's disease and intermediate-grade NHL are typical indications because of the requirement of accurate staging in determining the therapeutic protocol. One of the shortcomings of FDG-PET is its low specificity. Combination study with PET tracers with higher specificity in tumor diagnosis such as C-11 methionine or F-18 methyltyrosine could be a way to solve this problem.


The contribution of extrachromosomal DNA to genome plasticity in malaria parasites.
Malaria caused by the protozoan parasite Plasmodium falciparum continues to impose significant morbidity and mortality, despite substantial investment into drug and vaccine development and deployment. Underlying the resilience of this parasite is its remarkable ability to undergo genome modifications, thus, providing parasite populations with extensive genetic variability that accelerates selection of drug resistance and limits the efficacy of most vaccines. This genome plasticity is rooted in the mechanisms of DNA repair that parasites employ to maintain genome integrity, a process skewed toward homologous recombination through the evolutionary loss of classical nonhomologous end joining. Repair of DNA double-strand breaks have been shown to enable "shuffling" of antigen-encoding gene sequences to vastly increase antigen diversity and to enable copy number expansion of genes that contribute to drug resistance. The latter phenomenon has been proposed to be a major contributor to the rise of resistance to several classes of antimalarial drugs. In this issue of Molecular Microbiology, McDaniels and colleagues add yet another mechanism that malaria parasites use to reduce drug susceptibility by demonstrating that P. falciparum can maintain expanded arrays of drug resistance cassettes as stably replicating, circular, extrachromosomal DNAs, thus, expanding genome plasticity beyond the parasite's 14 nuclear chromosomes.


Evaluating stability of human spine in static tasks: a combined in vivo-computational study.
Various interpretations and parameters have been proposed to assess spinal stability such as antagonist muscle coactivity, trunk stiffness and spinal buckling load; however, the correlation between these parameters remains unknown. We evaluated spinal stability during different tasks while changing the external moment and load height and investigated likely relationships between different EMG- and model-based parameters (e.g., EMG coactivity ratio, trunk stiffness, force coactivity ratio) and stability margins. EMG and kinematics of 40 young healthy subjects were recorded during various quasi-static tasks. Muscle forces, trunk stiffness and stability margins were calculated by a nonlinear subject-specific EMG-assisted-optimization musculoskeletal model of the trunk. The load elevation and external moment increased muscle activities and trunk stiffness while all stability margins (i.e., buckling loads) decreased. The force coactivity ratio was strongly correlated with the hand-load stability margin (i.e., additional weight in hands to initiate instability; R2 = 0.54) demonstrating the stabilizing role of abdominal muscles. The total trunk stiffness (Pearson's r = 0.96) and the sum of EMGs of back muscles (Pearson's r = 0.65) contributed the most to the T1 stability margin (i.e., additional required load at T1 for instability/buckling). Force coactivity ratio and trunk stiffness can be used as alternative spinal stability metrics.


Self-Reported Executive Functioning in Everyday Life in Parkinson's Disease after Three Months of Subthalamic Deep Brain Stimulation.
Objective. Studies on the effect of subthalamic deep brain stimulation (STN-DBS) on executive functioning in Parkinson's disease (PD) are still controversial. In this study we compared self-reported daily executive functioning in PD patients before and after three months of STN-DBS. We also examined whether executive functioning in everyday life was associated with motor symptoms, apathy, and psychiatric symptoms. Method. 40 PD patients were examined with the Behavior Rating Inventory of Executive Function-Adult Version (BRIEF-A), the Symptom Checklist 90-Revised (SCL-90-R), and the Apathy Evaluation Scale (AES-S). Results. PD patients reported significant improvement in daily life executive functioning after 3 months of STN-DBS. Anxiety scores significantly declined, while other psychiatric symptoms remained unchanged. The improvement of self-reported executive functioning did not correlate with motor improvement after STN-DBS. Apathy scores remained unchanged after surgery. Only preoperative depressed mood had predictive value to the improvement of executive function and appears to prevent potentially favorable outcomes from STN-DBS on some aspects of executive function. Conclusion. PD patients being screened for STN-DBS surgery should be evaluated with regard to self-reported executive functioning. Depressive symptoms in presurgical PD patients should be treated. Complementary information about daily life executive functioning in PD patients might enhance further treatment planning of STN-DBS.


Leukemogenic risk of hydroxyurea therapy in polycythemia vera, essential thrombocythemia, and myeloid metaplasia with myelofibrosis.
In polycythemia vera (PV), treatment with chlorambucil and radioactive phosphorus (p32) increases the risk of leukemic transformation from 1% to 13-14%. This risk has been estimated to be 1-5.9% with hydroxyurea (HU) therapy. When compared with historical controls, the risk with use of HU does not appear to be statistically significant. The leukemogenic risk of HU therapy in essential thrombocytosis (ET) and in myelofibrosis with myeloid metaplasia (MMM) is unknown. HU remains the main myelotoxic agent in the treatment of PV, ET, and MMM. We studied 64 patients with these three disorders, seen at our institution during 1993-1995. The patients were studied for their clinical characteristics at diagnosis, therapies received, and development of myelodysplasia or acute leukemia (MDS/AL). Forty-two had PV, 15 ET, and 6 MMM, and 1 had an unclassified myeloproliferative disorder. Of the 42 patients with PV, 18 were treated with phlebotomy alone, 16 with HU alone, 2 with p32, 2 with multiple myelotoxic agents, and 2 with interferon-alpha (IFN-alpha). Two patients from the phlebotomy-treated group, one from the HU-treated group, and 1 from the multiple myelotoxic agent-treated group developed MDS/AL. In the larger group, 11 received no treatment or aspirin alone, 18 were treated with phlebotomy alone, 25 with HU, 5 with multiple myelotoxic agents, 2 with p32, 2 with IFN-alpha, and 1 with melphalan. Study of the entire group of 64 patients showed that only one additional patient (total of 5 out of 64) developed MDS/AL. This patient had been treated with HU alone. Statistical analysis did not show any association between clinical characteristics at diagnosis, or HU therapy, and development of MDS/AL (P=0.5). Thus, our data provide no evidence suggestive of increased risk of transformation to MDS/AL with HU therapy in PV, ET, and MMM. Larger, prospective studies are needed to study this issue further.


Social skills training for young adolescents: symbolic and behavioral components.
The assertiveness program developed by Wise, Bundy, Bundy, and Wise (1991), which focused on the unique problems and peer interactions of young adolescents, was expanded to a 12-week program and presented to a group of 28 sixth graders. Cognitive acquisition and retention of the symbolic information was measured with a pretest, posttest, and follow-up administration of the multiple-choice tests that were designed and used in the prior study. Verbal content of assertive behavior was measured in a pretest and posttest role-play situation. When trained students were compared to a control group on the symbolic measures, a significant effect for treatment (p < or = .000); a significant effect for measures (p < or = .000); and a significant Treatment x Measure interaction (p < or = .000) were found. A comparison of means showed a significant difference between pretest vs. posttest and follow-up (p < or = .000). However, no difference between posttest and follow-up was found (p < .351). Unlike the earlier study, in which girls benefited more from training than did boys, no treatment by gender interaction was found for the cognitive measure (p < .691). Boys and girls benefited similarly in terms of the cognitive acquisition of assertiveness information from this program. However, the results did not show that these students were able to demonstrate assertiveness on the behavioral level. Results are discussed in terms of the difference between simply having stored symbolic information regarding assertive responses and recognizing contexts in which accessing the information would be appropriate. Suggestions for designing programs aimed at bringing about changes in adolescents' assertive behavior that will generalize to their lives beyond the training context are also offered.


In vivo Application of the REMOTE-control System for the Manipulation of Endogenous Gene Expression.
Here we describe a protocol for implementing the REMOTE-control system (Reversible Manipulation of Transcription at Endogenous loci), which allows for reversible and tunable expression control of an endogenous gene of interest in living model systems. The REMOTE-control system employs enhanced lac repression and tet activation systems to achieve down- or upregulation of a target gene within a single biological system. Tight repression can be achieved from repressor binding sites flexibly located far downstream of a transcription start site by inhibiting transcription elongation. Robust upregulation can be attained by enhancing the transcription of an endogenous gene by targeting tet transcriptional activators to the cognate promoter. This reversible and tunable expression control can be applied and withdrawn repeatedly in organisms. The potency and versatility of the system, as demonstrated for endogenous Dnmt1 here, will allow more precise in vivo functional analyses by enabling investigation of gene function at various expression levels and by testing the reversibility of a phenotype.


Local and global existence theorems for the Einstein equations.
This article is a guide to the literature on existence theorems for the Einstein equations which also draws attention to open problems in the field. The local in time Cauchy problem, which is relatively well understood, is treated first. Next global results for solutions with symmetry are discussed. This is followed by a presentation of global results in the case of small data, and some miscellaneous topics connected with the main theme.


Temporal Change of Interleukin-6, C-Reactive Protein, and Skin Temperature after Total Knee Arthroplasty Using Triclosan-Coated Sutures.
The risk of surgical site infections (SSIs) after total knee arthroplasty (TKA) can never be eliminated. Antimicrobial sutures containing triclosan have been used to decrease SSIs, but whether triclosan-coated sutures are effective with TKA is unclear. Between 2011 and 2012, 102 patients randomly assigned to a triclosan or a control group were prospectively assessed. The incidence of SSI within 3 months of surgery, length of hospital stay, pain scale, functional scores, wound condition, and serum inflammatory markers during hospitalization and within 3 months postoperatively were compared. At the final follow-up, there were 2 patients with superficial infections (3.9%) in the control group but none in the triclosan group. Lower serum IL-6 was detected in the triclosan group at 4 weeks and 3 months. The local skin temperature of the knees-recorded at 3 months using infrared thermography-was lower in the triclosan group than in the control group. More precise analytical measurements are needed to investigate local and systemic complications, especially in the early subclinical stage. This prospective, randomized, open-label clinical trial is in the public registry: ClinicalTrials.gov (NCT02533492).


Transport and Compartmentation of 1-Aminocyclopropane-1-Carboxylic Acid and Its Structural Analog, alpha-Aminoisobutyric Acid, in Tomato Pericarp Slices.
The uptakes of 1-aminocyclopropane-1-carboxylic acid (ACC), the immediate precursor to ethylene, and its structural analog, alpha-aminoisobutyric acid (alphaAIB) by tomato pericarp slices were investigated. Both uptakes show a biphasic (saturable-linear) dependence on external concentration of the transported amino acid. At low concentrations, ACC uptake is competitively inhibited by alphaAIB and vice versa. Both uptakes also are inhibited by other neutral amino acids but not by acidic or basic amino acids. ACC and alphaAIB uptakes are metabolically dependent and are increased with time of tissue incubation. alphaAIB efflux patterns from pericarp slices indicated three distinct alphaAIB compartments having efflux kinetics consistent with those for cell wall, cytoplasm, and vacuole. The bulk of the alphaAIB taken up by pericarp tissue is sequestered into the vacuole. The ability of pericarp tissue to accumulate alphaAIB in the vacuole declines with fruit development.


Portal hypertension in lymphoproliferative and myeloproliferative disorders: hemodynamic and histological correlations.
The pathogenesis of portal hypertension in patients with lymphoproliferative and myeloproliferative disorders is not fully understood. We investigated 20 patients with myeloproliferative disease and 47 patients with lymphoproliferative disease. Transvenous liver biopsies and hepatic vein pressure gradient measurements were performed in all patients, and portal vein blood flow was measured by pulsed Doppler sonography in 31 of these patients and in 22 normal volunteers. The hepatic vein pressure gradient was significantly higher in patients with hepatic infiltrates, fibrosis or both than in patients without hepatic lesions (8.3 +/- 5.0 mmHg vs. 4.1 +/- 2.3 mmHg; p < 0.01). Portal vein blood flow was significantly higher in patients with hematological disease than in normal volunteers (31.2 +/- 15.5 ml/min.kg vs. 14.2 +/- 4.6 ml/min.kg;p < 0.01). In 81.8% of patients with hepatic infiltrates, fibrosis or both and increased portal vein blood flow, the hepatic vein pressure gradient was greater than 6 mm Hg. Although we saw a significant correlation between splenic vein blood flow and portal vein blood flow (n = 20; p < 0.01), we found no significant correlation between splenic vein blood flow and hepatic vein pressure gradient or spleen size. Hepatic infiltration and fibrosis appear to be major determinants of increased hepatic vein pressure gradient, probably because they increase intrahepatic vascular resistance. The role of increased splenic blood flow is probably not determinant. However, because portal pressure was not measured directly in this study, the incidence of portal hypertension may have been underestimated.


Sensitive quantitation of chromium-DNA adducts by inductively coupled plasma mass spectrometry with a direct injection high-efficiency nebulizer.
A novel method is described for the sensitive detection of chromium-DNA adducts. Chromium-DNA adducts were determined in 1 microgram of DNA from normal human lung fibroblasts exposed to sodium chromate using microscale flow injection analysis with a direct injection high-efficiency nebulizer and inductively coupled plasma mass spectrometry detection. The frequency of Cr-DNA adducts increased in a dose-dependent sigmoidal manner, indicating saturation and toxicity. The low detection limits (on the order of parts per trillion) allows the detection of as few as two Cr adducts per 10,000 bases, which, coupled with the small DNA sample requirement, makes this technique suitable for measuring metal-DNA adducts as biomarkers of exposure to toxic and carcinogenic metals such as Cr, in cultured cells, animals, and humans.


Phosphorylation of Thylakoid Proteins of Oryza sativa: In Vitro Characterization and Effects of Chilling Temperatures.
The phosphorylation of thylakoid proteins of rice (Oryza sativa L.) was studied in vitro using [gamma-(32)P]ATP. Several thylakoid proteins are labeled, including the light-harvesting complex of photosystem II. Protein phosphorylation is sensitive to temperature, pH, and ADP, ATP, and divalent cation concentrations. In the range pH 7 to 8.2, phosphorylation of the light-harvesting polypeptides declines above pH 7.5, whereas labeling of several other thylakoid polypeptides increases. Increasing divalent cation concentration from 3 to 20 millimolar results in a decrease in phosphorylation of the 26 kilodalton light-harvesting complex polypeptide and increased phosphorylation of several other polypeptides. ADP has an inhibitory effect on the phosphorylation of the light-harvesting complex polypeptides. Phosphorylation of the 26 kilodalton light-harvesting polypeptide requires 0.45 millimolar ATP for half-maximal phosphorylation, compared to 0.3 millimolar for the 32 kilodalton phosphoprotein. Low temperature inhibits the phosphorylation of thylakoid proteins in chilling-sensitive rice. However, phosphorylation of histones by thylakoid-bound kinase(s) is independent of temperature in the range of 25 to 5 degrees C, suggesting that the effect of low temperature is on accessibility of the substrate, rather than on the activity of the kinase.


Visceral fat area to appendicular muscle mass ratio as a predictor for nonalcoholic fatty liver disease independent of obesity.
Obesity and sarcopenia are known to be closely related to nonalcoholic fatty liver disease (NAFLD). We attempted to explore the combined influence of fat and muscle tissue on NAFLD by using visceral fat area to appendicular muscle mass ratio (VAR) as a novel parameter. In this cross-sectional study, a total of 3255 adults (1399 men and 1856 women) coming for a health examination were enrolled. NAFLD was diagnosed using ultrasound and VAR was measured by bioelectrical impedance analyzer. The prevalence of NAFLD was 46.5% in men and 26.6% in women. VAR differed significantly between subjects with and without NAFLD (4.27 vs. 3.26 in men, 7.89 vs. 5.01 in women, respectively, p < .001). Logistic regression analysis determined VAR as a risk factor for NAFLD, and the multivariable-adjusted odds ratios in the highest VAR quartile was 9.57 (95%CI: 5.98-15.30) for men and 12.37 (95%CI: 6.37-24.05) for women. From the receiver operating characteristic analysis, the area under the curve was 0.767 and 0.834, with the suitable cut-off VAR value of 3.469 and 6.357 for men and women, respectively. To control the influence of obesity, all subjects were stratified according to their BMI. For each BMI group, individuals with VAR above the cut-off value had significant higher prevalence and risk of NAFLD, with odds ratios ranging from 1.76 to 4.75. Increased VAR is strongly associated with higher risk of NAFLD in both sexes independent of obesity and can serve as a screening reference for NAFLD.


Equilibrium analysis of reactions of metal-pyridylazoresorcinolato chelates with quaternary ammonium ion, nonionic surfactant and polyethylene glycol in aqueous solution by capillary zone electrophoresis.
The resolutions of metal-4-(2-pyridylazo)resorcinol chelates by capillary zone electrophoresis (CZE) were investigated in the presence of some interacting reagents; also, equilibrium reactions between the chelates and the interacting reagents were analyzed in an aqueous solution. Among nine metal chelates formed in aqueous solution, the chelates of VV, FeII, CoIII, NiII, and CuII were resolved and detected by CZE, while other chelates were decomposed during electrophoretic migration. The electrophoretic mobility of the chelates of FeII, NiII, and CuII increased with increasing pH of the migrating solution; also, the acid-dissociation constants of these three chelates were determined by analyzing the mobility change. The ion-association constants of the five anionic chelates and pyridylazoresorcinolate ion with quaternary ammonium ions were also determined by analyzing the mobility change. The binding behavior of the ligand and its chelates with nonionic surfactant micelle, as well as with polyethylene glycol, were investigated, and their binding constants were determined through the mobility change. When Brij 35 was used as a nonionic surfactant interacting with the anionic chelates, the FeII chelate decomposed at Brij 35 concentrations over 6.67 mM. The equilibrium constants and the reactivity were compared with each other.


Effects of Environmental pH on the Internal pH of Chlorella pyrenoidosa, Scenedesmus quadricauda, and Euglena mutabilis.
The effect of external pH on two laboratory-cultured acid-intolerant species (Chlorella pyrenoidosa Chick and Scenedesmus quadricauda Turp. Bréb.) and one acid-tolerant species from a natural population (Euglena mutabilis Schmitz) was examined by measuring internal pH. These measurements were made with the weak acid (14)C-dimethyloxazolidine-2,4-dione after cells had been incubated for 2 and 6 hours at external pH levels from 3.0 to 8.0. Photosynthetic and respiration rates of the three species were also measured over the range of external pH levels.All three species regulated their internal pH levels over the 6-hour incubation time. C. pyrenoidosa and S. quadricauda had internal pH levels around 7.0, regardless of external pH. E. mutabilis had a wider internal pH range, from 5.0 at low external pH to 8.0 at high external pH. External pH had no effect on either photosynthetic or respiration rates. Statistical comparisons showed that there was a significant difference between the acid-intolerant and acid-tolerant species with regard to the level of internal pH maintained and the response of internal pH to external pH.


Comparative effectiveness of SGLT2i versus GLP1-RA on cardiovascular outcomes in routine clinical practice.
To investigate the comparative effectiveness of sodium-glucose cotransporter 2 inhibitors (SGLT2i) and glucagon-like peptide 1 receptor agonists (GLP1-RA) on cardiovascular outcomes in routine clinical practice, which have never been directly compared in head-to-head outcome trials. We compared outcomes of adults who newly started SGLT2i or GLP1-RA therapy in Stockholm, Sweden, during 2013-2019. The primary outcome was major adverse cardiovascular events (MACE), a composite of cardiovascular (CV) death, myocardial infarction and stroke. Secondary outcomes included the individual MACE components and hospitalization for heart failure. Cox regression with propensity score overlap weighting was used to estimate hazard ratios (HRs) with 95% confidence intervals and adjust for 57 covariates. We included 12,375 individuals, of which 5489 initiated SGLT2i and 6886 GLP1-RA therapy, followed for median 1.6 years. Mean age was 61 years and 37.6% were women. Compared with GLP1-RA, SGLT2i new users had similar risk of MACE risk (adjusted HR 1.04; 95% CI 0.83-1.31). The adjusted HRs (95% CI) for SGLT2i vs. GLP1-RA were 0.80 (0.59-1.09) for heart failure hospitalization, 0.95 (0.58-1.55) for cardiovascular death, 0.91 (0.67-1.24) for myocardial infarction and 1.71 (1.14-2.59) for ischemic stroke (5-year absolute risk difference for stroke 1.9% [95% CI 0.8-3.0]). In a largely primary-prevention population of people undergoing routine care, no differences were observed in MACE risk among initiators of SGLT2i and GLP1-RA. However, compared with GLP1RA, the use of SGLT2i was associated with an increased risk of ischemic stroke that was small in absolute magnitude.


nNOS-derived NO modulates force production and iNO-derived NO the excitability in C2C12-derived 3D tissue engineering skeletal muscle via different NO signaling pathways.
Nitric oxide (NO) is a bioactive gas produced by one of the three NO synthases: neuronal NOS (nNOS), inducible (iNOS), and endothelial NOS (eNOS). NO has a relevant modulatory role in muscle contraction; this takes place through two major signaling pathways: (i) activation of soluble guanylate cyclase and, thus, protein kinase G or (ii) nitrosylation of sulfur groups of cysteine. Although it has been suggested that nNOS-derived NO is the responsible isoform in muscle contraction, the roles of eNOS and iNOS and their signaling pathways have not yet been clarified. To elucidate the action of each pathway, we optimized the generation of myooids, an engineered skeletal muscle tissue based on the C2C12 cell line. In comparison with diaphragm strips from wild-type mice, 180 myooids were analyzed, which expressed all relevant excitation-contraction coupling proteins and both nNOS and iNOS isoforms. Along with the biochemical results, myooids treated with NO donor (SNAP) and unspecific NOS blocker (L-NAME) revealed a comparable NO modulatory effect on force production as was observed in the diaphragm strips. Under the effects of pharmacological tools, we analyzed the myooids in response to electrical stimulation of two possible signaling pathways and NO sources. The nNOS-derived NO exerted its negative effect on force production via the sGG-PKG pathway, while iNOS-derived NO increased the excitability in response to sub-threshold electrical stimulation. These results strengthen the hypotheses of previous reports on the mechanism of action of NO during force production, showed a novel function of iNOS-derived NO, and establish the myooid as a novel and robust alternative model for pathophysiological skeletal muscle research.


Synthesis of a new class of iminosugars based on constrained azaspirocyclic scaffolds by way of catalytic C-H amination.
The synthesis of the first examples of a new class of iminosugars based on constrained spirocyclic scaffolds has been achieved via Rh-catalyzed C(sp(3))-H amination. In this process, the needed electronic control in securing high regioselectivity from substrates with a high density of activated C-H bonds was achieved by using a combination of activating and electron-withdrawing groups.


Halogen-directed drug design for Alzheimer's disease: a combined density functional and molecular docking study.
A series of halogen-directed donepezil drugs has been designed to inhibit acetyl cholinesterase (AChE). Density Functional theory (DFT) has been employed to optimize the chair as well as boat conformers of the parent drug and modified ligands at B3LYP/MidiX and B3LYP/6-311G + (d,p) level of theories. Charge distribution, dipole moment, enthalpy, free energy and molecular orbitals of these ligands are also investigated to understand how the halogen-directed modifications impact the ligand structure and govern the non-bonding interactions with the receptors. Molecular docking calculation has been performed to understand the similarities and differences between the binding modes of unmodified and halogenated chair-formed ligands. Molecular docking indicated donepezil and modified ligands had non-covalent interactions with hydrophobic gorges and anionic subsites of AChE. The -CF3-directed ligand possessed the most negative binding affinity. Non-covalent interactions within the ligand-receptor systems were found to be mostly hydrophobic and π- stacking type. F, Cl and -CF3 containing ligands emerge as effective and selective AChE inhibitors, which can strongly interact with the two active sites of AChE. In addition, we have also investigated selected pharmacokinetic parameters of the parent and modified ligands.


Estimating weak ratiometric signals in imaging data. I. Dual-channel data.
Ratiometric fluorescent indicators are becoming increasingly prevalent in many areas of biology. They are used for making quantitative measurements of intracellular free calcium both in vitro and in vivo, as well as measuring membrane potentials, pH, and other important physiological variables of interest to researchers in many subfields. Often, functional changes in the fluorescent yield of ratiometric indicators are small, and the signal-to-noise ratio (SNR) is of order unity or less. In particular, variability in the denominator of the ratio can lead to very poor ratio estimates. We present a statistical optimization method for objectively detecting and estimating ratiometric signals in dual-wavelength measurements of fluorescent, ratiometric indicators that improves on standard methods. With the use of an appropriate statistical model for ratiometric signals and by taking the pixel-pixel covariance of an imaging dataset into account, we are able to extract user-independent spatiotemporal information that retains high resolution in both space and time.


Comparison of the behavioral and cardiovascular effects of intranasal and oral d-amphetamine in healthy human subjects.
Recent reports indicate an increase in intranasal use of prescription oral stimulant medication. However, there do not appear to be any published clinical studies that have characterized the behavioral and cardiovascular effects of intranasally administered d-amphetamine, which is commonly prescribed for Attention Deficit Hyperactivity Disorder. In this study, a range of d-amphetamine doses (0, 16, 24, and 32 mg/70 kg) were administered as an intranasal solution delivered using a mucosal atomization device. Equal oral doses were included for comparison. Assessments were conducted before and at regular intervals for 3 hours following drug administration and included self-reported drug-effect questionnaires, cardiovascular indices, a performance task, and 2 measures of impulsivity. d-Amphetamine produced prototypical stimulant effects (eg, increased subject ratings of Stimulated and Like Drug, elevated heart rate and blood pressure, and improved rate and accuracy on the digit symbol substitution task) irrespective of dose, but the onset of these effects was generally earlier following intranasal administration, with significant effects emerging 15 to 30 minutes after intranasal dosing and 45 to 60 minutes after oral dosing. These results demonstrate that intranasal administration of d-amphetamine results in a more rapid onset compared to oral dosing, which could be associated with the popularity of intranasal prescription stimulant use and an enhanced potential for abuse.


Rural nursing research priorities.
The magnitude and variety of clinical concerns in rural healthcare delivery mandate a focused plan for building a substantiated body of knowledge. The author identifies research priority areas to develop a body of knowledge for rural nursing.


Proteomic analysis reveals tanshinone IIA enhances apoptosis of advanced cervix carcinoma CaSki cells through mitochondria intrinsic and endoplasmic reticulum stress pathways.
Cervix cancer is the second most common cancer among women worldwide, whereas paclitaxel, the first line chemotherapeutic drug used to treat cervical cancer, shows low chemosensitivity on the advanced cervical cancer cell line. Tanshinone IIA (Tan IIA) exhibited strong growth inhibitory effect on CaSki cells (IC50 = 5.51 μM) through promoting caspase cascades with concomitant upregulating the phosphorylation of p38 and JNK signaling. Comprehensive proteomics revealed the global protein changes and the network analysis implied that Tan IIA treatment would activate ER stress pathways that finally lead to apoptotic cell death. Moreover, ER stress inhibitor could alleviate Tan IIA caused cell growth inhibition and ameliorate C/EBP-homologous protein as well as apoptosis signal-regulating kinase 1 mediated cell death. The therapeutic interventions targeting the mitochondrial-related apoptosis and ER stress responses might be promising strategies to conquer paclitaxel resistance.


[The use of electron microscopy for diagnosing spongiform encephalopathies].
The authors suggest an unsophisticated approach to laboratory diagnosis of spongiform encephalopathies induced by unconventional prion viruses, consisting in electron microscopic detection of abnormal helical Scrapie-associated fibrils. These fibrils are detectable in human and animal brain tissue samples and in cell cultures.


Respiratory effects of a single dive to 50 meters in sport divers with asymptomatic respiratory atopy.
Increasing popularity of sports diving makes it likely that subjects with allergic respiratory diseases will be involved in diving with self contained underwater breathing apparatus (scuba). The present study evaluated the effects of a single scuba-dive on pulmonary function in subjects with respiratory atopy. Specific airways conductance (sGaw), residual volume (RV), forced vital capacity (FVC), forced expiratory volume in 1 sec (FEV1), mid expiratory flow at 50% of FVC (MEF50), and transfer factor for carbon monoxide (TLCO) were measured in 9 sport divers with a history of hay fever and 9 matched healthy sport divers (control) before, 3 hours and 24 hours after a wet hyperbaric chamber dive to a depth of 50 m. Airway hyperresponsiveness (AHR) was assessed by methacholine challenge 4 weeks after the dive. Atopic subjects and controls did not differ with respect to anthropometric data, diving experience, and predive lung function. A 3% reduction in FVC was found 24h after the dive (p < 0.05) in both groups, whereas sGaw decreased by 15% 24 h after the dive (p < 0.05) in the subjects with respiratory atopy only. Postdive changes in RV, FEV1, MEF50, and TLCO did not reach level of statistical significance. AHR was obtained in 8/9 subjects with respiratory atopy. We conclude that subjects with atopic sensitization and asymptomatic AHR may be more susceptible to effects of diving on pulmonary function.


A mathematical model of a biological arms race with a dangerous prey.
In a recent paper, Brodie and Brodie provide a very detailed description of advances and counter-measures among predator-prey communities with a poisonous prey that closely parallel an arms race in modern society. In this work, we provide a mathematical model and simulations that provide a theory as to how this might work. The model is built on a two-dimensional classical predator-prey model that is then adapted to account for the genetics and random mating. The deterministic formulation for the genetics for the prey population has been developed and used in other contexts. Adapting the model to allow for genetic variation in the predator is much more complicated. The model allows for the evolution of the poisonous prey and for the evolution of the resistant predator. The biological paradigm is that of the poisonous newt and the garter snake which has been studied extensively although the models are broad enough to cover other examples.


Sex differences in gene expression related to antipsychotic induced weight gain.
Antipsychotics are crucial for the treatment of schizophrenia and contribute to weight gain in psychosis, particularly during early phases. Antipsychotic Induced Weight Gain (AIWG) might contribute to reduce the quality of life, drug compliance and to increase mortality. To characterize sex differences of gene expression related to AIWG, we sequenced total mRNA from blood samples of schizophrenia patients, before and after 3 months of antipsychotic-treatment. We analyzed schizophrenia patients according to their sex (38 males and 39 females) and their BMI increase after medication, characterizing the differential gene expression before and after medication. Individuals in each group were categorized in patients who gain weight and those whose do not gain weight. The "weight gain" groups included patients with an increase of body mass index (BMI) > 1.0 points (27 males and 23 females with a median BMI increase of 2.68 and 2.32 respectively). The "no weight gain" groups included patients with a change of BMI between < 1.0 and > -1.0 points (11 males and 16 females with a median BMI increase of 0.21 and 0.16 respectively). The males had 331 genes with significant differential expression in the weight gain group and 24 genes in the no weight gain group. The females had 119 genes with significant differential expression in the weight gain group and 75 genes in the no weight gain group. Both weight gain groups were significantly enriched with "obesity" genes (Fisher; p = 1.1E-09 and p = 0.0001 respectively), according to the Gene Reference into Function (GeneRIF) database.In conclusion, we characterized genes with differential expression associated to AIWG that are specific to males, to females and common to both sexes. These genes are good candidates to depict the biological processes involved in AIWG and provide additional evidence of the genetic links between weight gain and the immune system.


Neuropsychological intra-individual variability explains unique genetic variance of ADHD and shows suggestive linkage to chromosomes 12, 13, and 17.
Attention-deficit/hyperactivity disorder (ADHD) is a highly heritable neuropsychiatric disorder that is usually accompanied by neuropsychological impairments. The use of heritable, psychometrically robust traits that show association with the disorder of interest can increase the power of gene-finding studies. Due to the robust association of intra-individual variability with ADHD on a phenotypic and genetic level, intra-individual variability is a prime candidate for such an attempt. We aimed to combine intra-individual variability measures across tasks into one more heritable measure, to examine the relatedness to other cognitive factors, and to explore the genetic underpinnings through quantitative trait linkage analysis. Intra-individual variability measures from seven tasks were available for 238 ADHD families (350 ADHD-affected and 195 non-affected children) and 147 control families (271 children). Intra-individual variability measures from seven different tasks shared common variance and could be used to construct an aggregated measure. This aggregated measure was largely independent from other cognitive factors related to ADHD and showed suggestive linkage to chromosomes 12q24.3 (LOD = 2.93), 13q22.2 (LOD = 2.36), and 17p13.3 (LOD = 2.00). A common intra-individual variability construct can be extracted from very diverse neuropsychological tasks; this construct taps into unique genetic aspects of ADHD and may relate to loci conferring risk for ADHD (12q24.3 and 17p13.3) and possibly autism (12q24.3). Given that joining of data across sites boosts the power for genetic analyses, our findings are promising in showing that intra-individual variability measures are viable candidates for across site analyses where different tasks have been used.


The Partial Support of the Left Ventricular Assist Device Shifts the Systemic Cardiac Output Curve Upward in Proportion to the Effective Left Ventricular Ejection Fraction in Pressure-Volume Loop.
Left ventricular assist device (LVAD) has been saving many lives in patients with severe left ventricular (LV) failure. Recently, a minimally invasive transvascular LVAD such as Impella enables us to support unstable hemodynamics in severely ill patients. Although LVAD support increases total LV cardiac output (COTLV) at the expense of decreases in the native LV cardiac output (CONLV), the underlying mechanism determining COTLV remains unestablished. This study aims to clarify the mechanism and develop a framework to predict COTLV under known LVAD flow (COLVAD). We previously developed a generalized framework of circulatory equilibrium that consists of the integrated CO curve and the VR surface as common functions of right atrial pressure (PRA) and left atrial pressure (PLA). The intersection between the integrated CO curve and the VR surface defines circulatory equilibrium. Incorporating LVAD into this framework indicated that LVAD increases afterload, which in turn decreases CONLV. The total LV cardiac output (COTLV) under LVAD support becomes COTLV = CONLV+EFe · COLVAD, where EFe is effective ejection fraction, i.e., Ees/(Ees+Ea). Ees and Ea represent LV end-systolic elastance (Ees) and effective arterial elastance (Ea), respectively. In other words, LVAD shifts the total LV cardiac output curve upward by EFe · COLVAD. In contrast, LVAD does not change the VR surface or the right ventricular CO curve. In six anesthetized dogs, we created LV failure by the coronary ligation of the left anterior descending artery and inserted LVAD by withdrawing blood from LV and pumping out to the femoral artery. We determined the parameters of the CO curve with a volume-change technique. We then changed the COLVAD stepwise from 0 to 70-100 ml/kg/min and predicted hemodynamics by using the proposed circulatory equilibrium. Predicted COTLV, PRA, and PLA for each step correlated well with those measured (SEE; 2.8 ml/kg/min 0.17 mmHg, and 0.65 mmHg, respectively, r2; 0.993, 0.993, and 0.965, respectively). The proposed framework quantitatively predicted the upward-shift of the total CO curve resulting from the synergistic effect of LV systolic function and LVAD support. The proposed framework can contribute to the safe management of patients with LVAD.


Previous motor activity affects the transition from uncertainty to decision making in snails.
One of the most widely accepted benefits of enhanced physical activity is an improvement in the symptoms of depression, including the facilitation of decision making. Up until now, these effects have been shown in rodents and humans only. Little is known about their evolutionary origin or biological basis, and the underlying cellular mechanisms also remain relatively elusive. Here, we demonstrate for the first time that preceding motor activity accelerates decision making in an invertebrate, the pond snail Lymnaea stagnalis To investigate decision making in a novel environment, snails, which normally live in water, were placed on a flat dry surface to simulate the potentially threatening consequence of being in an arid environment. This stimulus initiated two distinct phases in snail behaviour: slow circular movements, followed by intense locomotion in a chosen direction. The first phase was prolonged when the test arena was symmetrically lit, compared with one with an apparent gradient of light. However, forced muscular locomotion for 2 h prior to the test promoted the transition from random circular motions to a directional crawl, accompanied by an increase in crawling speed but with no effect on the choice of direction. Intense locomotion for 2 h also produced a strong excitatory effect on the activity of serotonergic neurons in L. stagnalis Our results suggest that the beneficial effects of physical exercise on cognitive performance in mammals might have deep roots in evolution, granting the opportunity to unravel the origins of such effects at the single-neuron and network levels.


The common comorbidities leading to poor clinical outcomes after the surgical treatment of ankle fracture-dislocations.
The ankle fracture-dislocations are a significant traumatic incident for the bone and the soft tissue surrounding the ankle. Bone stabilization, joint immobilization, anatomic reduction and intervention for soft tissue protection should be performed as early as possible. The present study aims to determine the frequency of major comorbidities that can be seen after surgery in patients with ankle fracture-dislocations and the relationship between the trauma mechanism and clinical status with these comorbidities. Thirty-eight patients (25 males, 13 females) who underwent surgery with ankle fracture-dislocations between May 2014 and February 2017 were evaluated retrospectively in this study. All patients were evaluated clinically and radiologically at least 24 months postoperatively. Arthrosis, synostosis, presence of the chondral lesion and AOFAS scores were detected for all patients. Mean AOFAS score was lower in open ankle fracture-dislocations than in closed dislocations (p=0.044). An accompanying osteochondral lesion (OCL) and increased patient age were found to be strongly associated with the development of arthrosis (p=0.005 and p=0.017; respectively). Four of 29 patients who received primer definitive surgery and four of nine patients who received step-by-step surgery had poorly calculated AOFAS scores (p=0.071). There was no significant relationship between dislocation direction and AOFAS scores (p=0.087). Clinical and functional results were found to be worse in patients with open ankle fractures, the rate of arthrosis increased with age, and the use of syndesmosis screw had a positive but not a statistically significant effect on clinical and functional outcomes.


Experimentally-Induced Metabolic Acidosis Does not Alter Aortic Fatty Streak Formation in High-Cholesterol Fed Rabbits.
Cardiovascular disease causes a major clinical problem in patients with end stage renal disease. Since metabolic acidosis is very common in patients with end stage renal disease, we aimed to investigate the effect of experimentally-induced metabolic acidosis on serum lipid profile and aortic fatty streak (FS) formation in normal and high-cholesterol fed rabbits. Twenty-four male rabbits were divided into four groups (n=6 each): (1) normal diet (ND): (2) hypercholesterolemic diet (HCD) (1%): (3) ND plus acidemic diet: (4) HCD plus acidemic diet. Metabolic acidosis was induced by adding 0.75% NH4Cl in drinking water. After 4 weeks, blood samples were taken and thoracic aortae were dissected for histological examinations. Results showed that in the animals who received NH4Cl, metabolic acidosis was successfully induced. Serum total cholesterol and low density lipoprotein (LDL) concentrations in HCD groups were significantly higher than ND groups (P<0.05) and acidosis did not significantly change serum lipid levels neither in ND nor in HCD animals (P>0.05). Histological examination of aortae showed higher mean average grades of pathological evaluation in HCD than ND groups (2.1±0.16 vs. 0±0; P<0.05). Acidosis did not further increase FS formation in HCD groups (P >0.05). In this model of experimentally-induced metabolic acidosis, acidosis could not increase FS formation in HCD animals and it seems that it does not interfere in progression of atherosclerosis process.


Debatable donors: when can we count their consent?
Donor consent is the standard requirement for the acceptance of an organ or tissue donation from a living donor. Usually consent to donation is respected as the choice and act of an autonomous agent. There are times, however, when the consent should be paternalistically set aside. The problem for those who must make these decisions is determining when the donor's autonomy should be respected and when paternalistic interference is appropriate. This paper draws on the writings of Kant to develop an autonomy-preserving criterion for determining where to draw the line. It then goes on to display how that criterion could be applied to the case of an adolescent donor who agrees to donate a kidney to his brother.


Macular vessel density and foveal avascular zone parameters in patients after acute primary angle closure determined by OCT angiography.
This study analyzed the optical coherence tomography angiography (OCTA) macular parameters in primary angle-closure glaucoma (PACG) patients after acute primary angle closure (APAC) episodes. Thirty-three patients with 33 APAC eyes and 33 primary angle closure suspect (PACS) eyes and 33 age-matched normal subjects (controls) were enrolled. Macular vessel density (VD) in central, inner, outer and full regions and foveal avascular zone (FAZ) parameters (area, perimeter and circularity index) were compared between APAC, PACS, and control eyes. For resolved APAC eyes, the VD in each macular region was significantly lower than that in control eyes, with less central and inner macular VD than PACS eyes. The central macular VD was significantly lower in PACS eyes than in controls. There was no difference in FAZ area and perimeter between APAC, PACS, and control eyes. FAZ circularity was highest in control eyes, followed by PACS eyes, and lowest in APAC eyes. The AUC, sensitivity and specificity of FAZ circularity were 0.944, 93.9% and 84.8%, respectively, in APAC eyes and 0.881, 84.8% and 81.8%, respectively, in PACS eyes. Therefore, FAZ circularity had the best discrimination capability for detecting both APAC and PACS eyes. Macular assessment with OCTA could provide an accurate early-stage diagnostic tool for PACG.


A 3-dimensional finite element analysis to evaluate the impact of force direction, insertion angle, and cortical bone thickness on mini-screw and its surrounding bone.
Background. The present study aimed to assess the stress and strain distribution on mini-screws and the surrounding bone in cases of different cortical bone thicknesses (CBTs), mini-screw insertion angles, and force directions using finite element analysis (FEA). Methods. Inventor professional version 8 software was used to construct 24 three-dimensional assemblies of mini-screws inserted with different insertion angles (30º, 60º, and 90º) in alveolar bone blocks with different CBTs (0.5, 1, 1.5, and 2 mm). The models simulated mini-screws inserted in bones with different CBTs and different insertion angles. A 2-N load was applied in two directions to mini-screw heads. The resultant stresses of the applied load were collected from the output of the ANSYS program. Results. The results indicated that force direction affected bone strains as the horizontal force generated more strains on cortical bone than the oblique one. Force applied to 60º inserted mini-screws generated much more strains on cortical bone than 90º and 30º inserted mini-screws. In a 60º inserted mini-screw, the horizontal force generated about 45% more strains on cortical bone than the oblique one. The exerted microstrain on bone decreased as CBT increased. Conclusion. It can be concluded that inserting mini-screws at 60º to the bone surface should be avoided as it generates much more strains on cortical bone than 90º and 30º, especially when a force parallel to the bone surface is applied.


Canada on the Move: an intensive media analysis from inception to reception.
Research evaluating mediated physical activity campaigns uses an unsophisticated conceptualization of the media and would benefit from the application of a media studies approach. The purpose of this article is to report on the application of this type of analysis to the Canada on the Move media campaign. Through interviews and document analysis, the press release surrounding Canada on the Move was examined at four levels: inception, production, transmission and reception. Analytic strategies of thematic and textual analysis were conducted. The press release was well received by journalists and editors and was successfully transmitted as inferred from national and local television coverage, although there was no national print pickup. Canada on the Move was perceived by sampled audience members as a useful and interesting strategy to encourage walking. A holistic approach to media analysis reveals the complex and frequently messy process of this mediated communication process. Implications for future media disseminations of Canada on the Move are discussed.


Phase I-II study of dose intensified chemotherapy with filgrastim and thymopentin in patients with advanced cancer.
Carboplatin (CBDCA), cyclophosphamide (CTX) and etoposide (VP-16) combination chemotherapy is active in many tumors. A phase I-II study was designed in order to verify toxicity, maximum tolerated dose and activity of CBDCA, CTX and VP-16 given with Granulocyte Colony Stimulating Factor (G-CSF) and thymopentin (TP5), without bone marrow support. A group of 12 heavily pretreated patients (PTS), 9 breast cancer, 2 small cell lung cancer and 1 gastric cancer, received fourteen courses of the described combination chemotherapy. Previous treatments were as follows: surgery in 10 PTS, radiotherapy in 7 PTS, chemotherapy in all PTS (median of 9 courses per patient). CBDCA, CTX, VP-16 were given over 3 days. CBDCA doses were calculated and expressed with the area under the concentration versus time curve (AUC). The dose range were as follows: CBDCA AUC 5.5-11 (400-800 mg/m2), CTX 1500-2500 mg/m2, VP-16 450-550 mg/m2, G-CSF was given 5 mg/kg/day from day 4 to day 17. TP5 was given, on alternate days, 1 mg/ kg from day 4 to day 30. 6 PTS developed fever > 38 degrees C for a median (M) duration of 4 days. 4 PTS required platelet support (M of 12 units) and 3 PTS red blood cells support (M of 2 units). Maximum tolerated dose was CBDCA AUC of 8, CTX 2000 mg/m2 and VP16500 mg/m2. No treatment related death occurred. Ten patients had responses and two had disease stabilisation. At the present time 5 PTS are alive and median overall survival is 13 months. These data indicate that dose intensified CT with the support of G-CSF and TP5 may be delivered safely and shows activity in heavily pre-treated patients.


[Virtual colonoscopy in stenosing colorectal cancer].
In 5-10% of patients it is not possible to achieve a complete endoscopic examination of the colon, because of obstructing cancer, excessive length of the colon, anatomical abnormalities or adhesions. Virtual colonoscopy is currently capable of investigating the colic lumen with a non-invasive technique, with high specificity and sensitivity. From January 2005 to July 2007 we treated 21 patients with obstructing neoplastic colorectal lesions, preventing a complete endoscopic examination. In all patients we performed a virtual colonoscopy, which revealed the presence of synchronous lesions (19%): a pedunculated polyp in two cases, a sessile polyp in one case and a right colonic vegetating lesion. In the 21 patients studied we performed a follow-up colonoscopy 3 months after the surgical treatment. No other endoluminal lesions were found, confirming the results of virtual colonoscopy. In our experience virtual colonoscopy presented 100% sensitivity and specificity. In this selected group of patients with obstructing lesions of the colon, virtual colonoscopy enables the surgeon to evaluate the entire colon, avoiding the execution of an intraoperative colonoscopy and possible surgical reintervention due to the finding of synchronous neoplastic lesions at postoperative follow-up endoscopy.


Exposure to humidifier disinfectants induces developmental effects and disrupts thyroid endocrine systems in zebrafish larvae.
Humidifier disinfectants have been widely used in Korea to prevent the growth of microorganisms in humidifier water. However, their use has been banned since 2011 after epidemiological studies reported humidifier disinfectant induced lung injury. In the present study, the developmental effects of exposure to two humidifier disinfectants (Oxy® and Wiselect) and their main component, polyhexamethylene guanidine (PHMG)-phosphate, were investigated in zebrafish embryos/larvae for seven days. The effects on triiodothyronine (T3) and thyroxine (T4) hormones, reactive oxygen species (ROS) generation, antioxidant enzyme activities, and changes in expression of the genes related to the hypothalamus-pituitary-thyroid (HPT) axis and oxidative stress were also investigated. Zebrafish embryos exposed to the highest concentration (amounts recommended for use by the manufacturers) of all tested humidifier disinfectants showed an increase in embryo coagulation, leading to death without hatching. Exposure to Oxy® and Wiselect resulted in significantly decreased body length, increased ROS generation and antioxidant enzyme activities, decreased T4, and up-regulated genes related to the HPT axis (trh, trβ, and tpo) and oxidative damage (sod2 and gpx1b). The humidifier disinfectants and PHMG-phosphate could induce oxidative stress and disrupt thyroid hormone systems in zebrafish, leading to developmental retardation when used at sub-lethal concentrations. Potential effects of long-term exposure to humidifier disinfectants and mixture effects of several major components deserve further investigation.


Pituitary hormone gene expression and secretion in human growth hormone-releasing hormone transgenic mice: focus on lactotroph function.
The human GH-releasing hormone (hGHRH) transgenic mouse has a hyperplastic anterior pituitary gland that eventually develops into an adenoma. We showed previously that the number of lactotrophs in the male hGHRH transgenic mouse is increased 2-fold, yet there is no concomitant increase in plasma levels of PRL. To further elucidate underlying changes in lactotroph function in the hGHRH transgenic mouse, the objectives of this study were to 1) examine the relative differences in PRL gene expression in transgenic mice and their siblings, 2) quantify PRL secretion at the level of the individual cell, 3) determine whether tyrosine hydroxylase gene expression and/or activity are altered in the hypothalamus of transgenic mice, and 4) assess dopamine receptor gene expression and functional sensitivity in lactotrophs of transgenic mice. Total PRL messenger RNA (mRNA) levels were increased nearly 5-fold in the hGHRH transgenic mouse, whereas the concentrations of PRL mRNA (PRL mRNA per microg total RNA) were unchanged. In contrast, total PRL contents were unchanged, whereas the concentrations of PRL (micrograms of PRL per mg total protein) were decreased 3-fold. Hypothalamic tyrosine hydroxylase steady state mRNA levels were not altered in the hGHRH transgenic mice, but hypothalamic tyrosine hydroxylase activity was increased 2-fold in transgenic mice. Dopamine D2 receptor mRNA concentrations in the anterior pituitary were increased 2.5-fold in hGHRH transgenic mice, and total pituitary D2 receptor mRNA levels were increased nearly 10-fold. Furthermore, the basal secretory capacity of lactotrophs from transgenic mice was increased significantly at the level of the single cell, and dopamine inhibited the secretion of PRL to a greater extent in hGHRH transgenic mice. Thus, although the total number of lactotrophs is increased 2-fold in hGHRH transgenic mice, the present data are consistent with the hypothesis that increased hypothalamic dopamine synthesis and release coupled with an increase in D2 dopamine receptor gene expression and functional sensitivity in the pituitary result in normal plasma levels of PRL.


Community-based dental services for patients with special needs.
Deinstitutionalization of individuals with mental retardation and developmental disabilities has increased the demand for dental services for these patients by community practitioners. There are numerous difficulties associated with the delivery of care to this population with special needs. Nevertheless, a county-by-county review for New York State indicates that in most instances, if all dentists are willing to help, there would be a relatively small number of these patients per dental practitioner.


Efficient workflow for the investigation of the catalytic cycle of water oxidation catalysts: Combining GFN-xTB and density functional theory.
Photocatalytic water oxidation remains the bottleneck in many artificial photosynthesis devices. The efficiency of this challenging process is inherently linked to the thermodynamic and electronic properties of the chromophore and the water oxidation catalyst (WOC). Computational investigations can facilitate the search for favorable chromophore-catalyst combinations. However, this remains a demanding task due to the requirements on the computational method that should be able to correctly describe different spin and oxidation states of the transition metal, the influence of solvation and the different rates of the charge transfer and water oxidation processes. To determine a suitable method with favorable cost/accuracy ratios, the full catalytic cycle of a molecular ruthenium based WOC is investigated using different computational methods, including density functional theory (DFT) with different functionals (GGA, Hybrid, Double Hybrid) as well as the semi-empirical tight binding approach GFN-xTB. A workflow with low computational cost is proposed that combines GFN-xTB and DFT and provides reliable results. GFN-xTB geometries and frequencies combined with single-point DFT energies give free energy changes along the catalytic cycle that closely follow the full DFT results and show satisfactory agreement with experiment, while significantly decreasing the computational cost. This workflow allows for cost efficient determination of energetic, thermodynamic and dynamic properties of WOCs.


Stereoselective synthesis of cycloheptanone derivatives via an intermolecular [5 + 2] cycloaddition reaction.
[reaction: see text] A [5 + 2] cycloaddition reaction of a new five-carbon unit was developed on the basis of a dicobalt hexacarbonyl propargyl cation species. Under the influence of EtAlCl(2), [5-benzoyloxy-2-(triisopropylsiloxy)-1-penten-3-yne)]dicobalt hexacarbonyl reacted with enol triisopropylsilyl ethers to yield seven-membered dicobalt acetylene complexes in good yield. The reactions with cyclic enol silyl ethers as well as acyclic enol silyl ethers exhibited remarkably high diastereoselectivity. The cycloadducts can be easily converted into various kinds of cycloheptanone derivatives.


Interface between adaptive and neoplastic growth in the pancreas.
The adaptive changes of hypertrophy and hyperplasia are diffuse and reversible responses of the pancreas to growth promoting stimuli. Early stages of neoplastic growth in the pancreas have been studied in carcinogen treated animals and preneoplastic lesions including atypical acinar cell foci and nodules, tubular ductal complexes and intraductal hyperplasia were identified. Neoplastic growth is clonal rather than diffuse and involves multiple steps through preneoplastic stages to produce a tumour. The individual steps are commonly regarded as reflecting a series of changes in the genome of the cells. Although the changes are likely to be irreversible, completion of the sequence usually requires a major portion of the lifespan of the host. The rate of progression of preneoplastic lesions to cancer may be modulated by the same factors that control adaptive growth. It follows that such factors will influence the probability that a carcinoma will develop. Cholecystokinin (CCK) seems to provide one example of a hormone/growth factor that can stimulate normal, adaptive, and neoplastic growth, and it is to be expected that other such hormones will be identified.