Modulation of Biomarker Expression by Osimertinib: Results of the Paired Tumor Biopsy Cohorts of the AURA Phase I Trial.
Osimertinib is an oral, potent, irreversible EGFR tyrosine kinase inhibitor (TKI) selective for EGFR TKI and T790M resistance mutations. To enhance understanding of osimertinib's mechanism of action, we aimed to evaluate the modulation of key molecular biomarkers after osimertinib treatment in paired clinical samples from the phase I AURA trial. Paired tumor biopsy samples were collected before the study and after 15 plus or minus 7 days of osimertinib treatment (80 or 160 mg daily). Clinical efficacy outcomes were assessed according to whether viable paired biopsy samples could be collected; safety was also assessed. Immunohistochemical analyses assessed key pathway and tumor/immune-relevant markers (phospho-EGFR, phospho-S6, phospho-AKT, programmed death ligand 1, and CD8), with samples scored by image analysis or a pathologist blinded to treatment allocation. Predose tumor biopsy samples were collected from 61 patients with EGFR T790M tumors; 29 patients had no viable postdose biopsy sample because of tumor regression or insufficient tumor sample. Evaluable predose and postdose tumor biopsy samples were collected from 24 patients. Objective response rate (ORR) and median progression-free survival (mPFS) were improved in patients from whom a postdose biopsy sample could not be collected (ORR 62% and mPFS 9.7 months [p = 0.027]) compared with those from whom paired samples were collected (ORR 29% and mPFS 6.6 months). Osimertinib modulated key EGFR signaling pathways and led to increased immune cell infiltration. Collection of paired biopsy samples was challenging because of rapid tumor regression after osimertinib treatment, highlighting the difficulties of performing on-study biopsies in patients treated with highly active drugs.


Implementing methods in the ELEGANCE registry to increase diversity in clinical research.
Women and underrepresented minorities (URMs) who are at an increased risk of presenting with severe peripheral artery disease (PAD) and have different responses to treatment compared with non-Hispanic White males yet are underrepresented in PAD research. ELEGANCE is a global, prospective, multi-center, post-market registry of PAD patients treated with drug-eluting device that aims to enroll at least 40% women and 40% URMs. The study design incorporates strategies to increase enrollment of women and URMs. Inclusion criteria are age ≥18 years and treatment with any commercially available Boston Scientific Corporation drug-eluting device marketed for peripheral vasculature lesions; exclusion criterion is life expectancy <1 year. Of 750 patients currently enrolled (951 lesions) across 39 sites, 324 (43.2%) are female and 350 (47.3%) are URMs (21.6% Black, 11.2% Asian, 8.5% Hispanic/Latino, and 5.3% other). Rutherford classification is distributed differently between sexes (P = .019). Treatment indication differs among race/ethnicity groups (P = .003). Chronic limb-threatening ischemia was higher for Black (38.3%) and Hispanic/Latino (28.1%) patients compared with non-Hispanic White (21.8%) and Asian patients (21.4%). De-novo stenosis was higher in Asian patients (92.3%) compared with Black, non-Hispanic White, and Hispanic/Latino patients (72.2%, 68.7%, and 77.8%, respectively; P < .001). Mean lesion length was longest for Black patients (162.7 mm), then non-Hispanic White (135.2 mm), Asian (134.8 mm), and Hispanic/Latino patients (128.1 mm; P = .008). Analyses of data from the ELEGANCE registry show that differences exist in baseline disease characteristics by sex and race/ethnicity; these may be the result of other underlying factors, including time to diagnosis, burden of undermanaged comorbidities, and access to care.


Hippocampal cytogenesis abrogation impairs inter-regional communication between the hippocampus and prefrontal cortex and promotes the time-dependent manifestation of emotional and cognitive deficits.
Impaired ability to generate new cells in the adult brain has been linked to deficits in multiple emotional and cognitive behavioral domains. However, the mechanisms by which abrogation of adult neural stem cells (NSCs) impacts on brain function remains controversial. We used a transgenic rat line, the GFAP-Tk, to selectively eliminate NSCs and assess repercussions on different behavioral domains. To assess the functional importance of newborn cells in specific developmental stages, two parallel experimental timeframes were adopted: a short- and a long-term timeline, 1 and 4 weeks after the abrogation protocol, respectively. We conducted in vivo electrophysiology to assess the effects of cytogenesis abrogation on the functional properties of the hippocampus and prefrontal cortex, and on their intercommunication. Adult brain cytogenesis abrogation promoted a time-specific installation of behavioral deficits. While the lack of newborn immature hippocampal neuronal and glial cells elicited a behavioral phenotype restricted to hyperanxiety and cognitive rigidity, specific abrogation of mature new neuronal and glial cells promoted the long-term manifestation of a more complex behavioral profile encompassing alterations in anxiety and hedonic behaviors, along with deficits in multiple cognitive modalities. More so, abrogation of 4 to 7-week-old cells resulted in impaired electrophysiological synchrony of neural theta oscillations between the dorsal hippocampus and the medial prefrontal cortex, which are likely to contribute to the described long-term cognitive alterations. Hence, this work provides insight on how newborn neurons and astrocytes display different functional roles throughout different maturation stages, and establishes common ground to reconcile contrasting results that have marked this field.


[Estradiol inhibits dopamine mediated behavior in rats--an animal model of sex-specific differences in schizophrenia].
We found in a representative sample of 392 first hospital admissions for schizophrenia a higher mean age at onset in females by 3.2 to 3.9 years, whereas the lifetime risk was equal for both sexes. In males the rates of onset show a steep increase reaching the maximum value in the age group 15-24 years, followed then by a steady decrease. Females reach the first peak with a clear delay between 20 and 29 years. After the decrease a second smaller peak is observed consistently in females within the age group 45-49 years and over. After having excluded alternative explanations for this gender differences (for example, diagnosis artefacts, sociocultural factors), we hypothesized that the effect of oestradiol on the dopaminergic system enhances the vulnerability threshold for schizophrenia, which is lowered again during the menopause. Alternatively we assumed that testosterone reduces the vulnerability threshold and thus furthers the earlier onset of schizophrenia in males. We tested these hypotheses in animal models by investigating the effects of the gonadal hormones on haloperidol-induced catalepsy and on apomorphine-induced stereotypies in both neonatal and adult rats. Testosterone showed no clear influence on the tested dopamine-mediated behaviour. Oestradiol caused a significant reduction on both dopamine-agonist and dopamine-antagonist induced behaviour. These effects were stronger in neonatal animals. Since oestradiol caused a 2.8-fold reduction of dopamine receptor affinity for sulpiride, we assumed that the behavioural changes caused by oestradiol were accounted for by a down-regulation of the dopaminergic system.(ABSTRACT TRUNCATED AT 250 WORDS)


Molecular cloning of the cowpea leghemoglobin II gene and expression of its cDNA in Escherichia coli. Purification and characterization of the recombinant protein.
Cowpea (Vigna unguiculata) nodules contain three leghemoglobins (LbI, LbII, and LbIII) that are encoded by at least two genes. We have cloned and sequenced the gene that encodes for LbII (lbII), the most abundant Lb in cowpea nodules, using total DNA as the template for PCR. Primers were designed using the sequence of the soybean lbc gene. The lbII gene is 679 bp in length and codes for a predicted protein of 145 amino acids. Using sequences of the cowpea lbII gene for the synthesis of primers and total nodule RNA as the template, we cloned a cDNA for LbII into a constitutive expression vector (pEMBL19+) and then expressed it in Escherichia coli. Recombinant LbII (rLbII) and native LbII (nLbII) from cowpea nodules were purified to homogeneity using standard techniques. Properties of rLbII were compared with nLbII by partially sequencing the proteins and by sodium dodecyl sulfate- and isoelectric focusing polyacrylamide gel electrophoresis, western-blot analysis using anti-soybean Lba antibodies, tryptic and chymotryptic mapping, and spectrophotometric techniques. The data showed that the structural and spectral characteristics of rLbII and nLbII were similar. The rLbII was reversibly oxygenated/deoxygenated, showing that it is a functional hemoglobin.


Inhibition of cell migration and invasion by miR‑29a‑3p in a colorectal cancer cell line through suppression of CDC42BPA mRNA expression.
The objective of this study was to determine the effect of miR‑29a‑3p inhibitor on the migration and invasion of colorectal cancer cell lines (CRC) and the underlying molecular mechanisms. miR‑29a‑3p was detected using reverse transcription-quantitative polymerase chain reaction (RT‑qPCR) in the CRC cell lines HCT11, CaCo2, HT29, SW480 and SW620. An invasive subpopulation designated SW480‑7 was derived from the parental cell line, detected by Transwell and Transwell Matrigel assays. Cytoskeleton Regulators RT2 profiler PCR array and western blot analysis were utilized to identify the alterations in expression of downstream mRNAs. siRNA against CDC42BPA was transfected into SW480‑7 and effects on cell migration and invasion were investigated. Data obtained showed that miR‑29a‑3p was detected in these five CRC cell lines. miR‑29a‑3p inhibitor had no effect on viability but stimulated cell migration and invasion of SW480‑7 cells. In contrast, miR‑29a‑3p mimic suppressed cell migration and invasion. TargetScan miRBD and DIANA were employed to identify the potential direct target genes of miR‑29a‑3p in the Cytoskeleton Regulators RT2-Profiler PCR array. Cytoskeleton Regulators RT2-Profiler PCR array data showed that 3 out of the 5 predicted targets genes, CDC42BPA (2.33-fold), BAIAP2 (1.79-fold) and TIAM1 (1.77-fold), in the array were upregulated by miR‑29a‑3p. A significant increase in expression IQGAP2, PHLDB2, SSH1 mRNAs and downregulation of PAK1 mRNA was also detected with miR‑29a‑3p inhibition. Increase in CDC42BPA, SSH1 and IQGAP2 mRNA expression correlated with increased protein level in miR‑29a‑3p transfected SW-480-7 cells. Silencing of CDC42BPA (an enhancer of cell motility) partially abolished miR‑29a‑3p inhibitor-induced stimulation of cell migration and invasion. miR‑29a‑3p expression in stage II and III CRC is relatively lower than that of stage I CRC. However, the data need to be interpreted with caution due to the small sample size. In conclusion, inhibition of miR‑29a‑3p stimulates SW480‑7 cell migration and invasion and downstream expression IQGAP2, PHLDB2, SSH1 mRNAs are upregulated whilst PAK1 mRNA is downregulated. Silencing of CDC42BPA expression partially reduces miR29a‑3p inhibitor-induced migration and invasion of SW480‑7 cells.


Reversal of cannabinoids (delta9-THC) by the benzoflavone moiety from methanol extract of Passiflora incarnata Linneaus in mice: a possible therapy for cannabinoid addiction.
The newly reported benzoflavone moiety from the plant Passiflora incarnata Linneaus has been evaluated in light of traditional reports on the use of P. incarnata in breaking down cannabis addiction. In the modern or allopathic system of therapeutics, there has been no suitable remedy to combat the severe withdrawal effects of various cannabis products, including marihuana, marijuana, bhang, hashish, ganja, etc., the world-wide consumption of which has attained alarming proportions especially among the younger generation. Mice were given a 10-mg-kg(-1) twice-daily dose of delta9-tetrahydrocannabinol (delta9-THC) by mouth for six days to make them dependent upon cannabinoids. Concurrently, other groups of mice were administered delta9-THC along with a 10- or 20-mg-kg(-1) twice-daily dose of the benzoflavone moiety from P. incarnata orally for 6 days. Upon measuring locomotor activity during the treatment regimen, it was noticed that the mice receiving the P. incarnata extract and delta9-THC together developed significantly less tolerance and dependence, relative to the mice receiving delta9-THC alone. Upon administration of SR-141716A, a selective cannabinoid-receptor antagonist (10 mg kg(-1), p.o.) to all the groups of mice on the 7th day, an artificial withdrawal was produced due to an abrupt decline of delta9-THC levels in mouse brain. However, the typical withdrawal effects like paw tremors and head shakes were significantly less in the mice given delta9-THC+P. incarnata benzoflavone moiety for 6 days. Upon administration of 20 mg kg(-1) of the P. incarnata benzoflavone moiety to mice showing severe symptoms of withdrawal due to administration of SR-141716A, there was a marked attenuation of withdrawal effects, thereby suggesting the usefulness of the benzoflavone moiety in delta9-THC withdrawal. Thus, the benzoflavone moiety of P. incarnata, when administered concurrently with delta9-THC, prevented the development of tolerance and dependence of cannabinoids in mice. Even an acute administration of the benzoflavone moiety (20 mg kg(-1), p.o.) significantly blocked the expression of withdrawal effects in delta9-THC-dependent mice.


Hck/Fgr Kinase Deficiency Reduces Plaque Growth and Stability by Blunting Monocyte Recruitment and Intraplaque Motility.
Leukocyte migration is critical for the infiltration of monocytes and accumulation of monocyte-derived macrophages in inflammation. Considering that Hck and Fgr are instrumental in this process, their impact on atherosclerosis and on lesion inflammation and stability was evaluated. Hematopoietic Hck/Fgr-deficient, LDLr(-/-) chimeras, obtained by bone marrow transplantation, had smaller but, paradoxically, less stable lesions with reduced macrophage content, overt cap thinning, and necrotic core expansion as the most prominent features. Despite a Ly6C(high)-skewed proinflammatory monocyte phenotype, Hck/Fgr deficiency led to disrupted adhesion of myeloid cells to and transmigration across endothelial monolayers in vitro and atherosclerotic plaques in vivo, as assessed by intravital microscopy, flow cytometry, and histological examination of atherosclerotic arteries. Moreover, Hck/Fgr-deficient macrophages showed blunted podosome formation and mesenchymal migration capacity. In consequence, transmigrated double-knockout macrophages were seen to accumulate in the fibrous cap, potentially promoting its focal erosion, as observed for double-knockout chimeras. The hematopoietic deficiency of Hck and Fgr led to attenuated atherosclerotic plaque formation by abrogating endothelial adhesion and transmigration; paradoxically, it also promoted plaque instability by causing monocyte subset imbalance and subendothelial accumulation, raising a note of caution regarding src kinase-targeted intervention in plaque inflammation.


A structural equation modeling analysis of the effects of nurses' spirituality and spiritual care on professional quality of life.
Spiritual care helps nurses establish a deeper connection with patients and meet their spiritual needs. Spiritual belief is thought to enable nurses to be more effective in their profession and positively affect their quality of life. This study aimed to investigate the effects of nurses' spirituality and spiritual care on quality of life. This study was designed as a descriptive cross-sectional study using structural equation modeling. A total of 221 nurses were included. Data were collected using the Professional Quality of Life Scale, and Spirituality and Spiritual Care Scale. The data were analyzed using descriptive statistics, correlational statistics, and structural equation modeling. Spirituality and spiritual care were negative predictors of burnout and positive predictors of compassion satisfaction. Spirituality and spiritual care decreased the level of burnout in nurses and significantly increased the level of compassion satisfaction. This study suggests raising nurse awareness of spirituality and spiritual care. Supporting nurses with professional training programs, including spiritual care, may benefit their quality of life.


Prospective audit of febrile neutropenia management at a tertiary university hospital in Singapore.
Febrile neutropenia (FN) remains a major cause of morbidity and mortality in Oncology/Haematology units. We launched a new protocol for FN management that incorporates risk stratification at our institute from October 2008. An audit was performed concurrently to evaluate the protocol and to define the epidemiology of FN locally. Case records of all inpatients with FN between October 2008 and June 2009 were reviewed prospectively. Clinical and microbiological characteristics were collated along with outcomes and programme adherence. Statistical testing was performed using Stata 10.1. There were 178 FN episodes (50 in patients with solid cancers) from 131 patients. Forty-two (23.6%) episodes were classified as high-risk according to MASCC criteria. Initial blood cultures were positive in 49 (27.5%) episodes, of which gram-negative bacilli (GNB) predominated. Overall compliance to the protocol was 56.7%, with the main issue being disinclination to use oral antibiotics as fi rst-line empirical therapy for low-risk episodes. Overall mortality was 7.3% and infection-related mortality was 4.5%. High-risk FN and the presence of central venous catheters were independently associated with bacteraemia on multivariate analysis, but there were no independent predictors of infection-related mortality. GNB accounted for the majority of bloodstream infections at our institute, unlike data from developed countries. Uptake of the new FN protocol was satisfactory, although the use of oral antibiotics as fi rst-line empirical therapy can be improved. A better method for predicting infections caused by antibiotic-resistant GNB is urgently required, and antibiotic resistance trends should be monitored to enable the implementation of more appropriate antibiotic regimens over time.


Intraoperative ultrasonography with a surgeon-manipulated microtransducer during robotic radical prostatectomy.
Intraoperative transrectal ultrasonography during laparoscopic radical prostatectomy has been reported to lead to a reduction in surgical margin rates. However, the use of a surgeon-controlled ultrasound probe that allows for precise manipulation and direct interpretation of the image by a console surgeon has yet to be studied. The aim of the present study was to show initial feasibility using the microtransducer with 9-mm scan length controlled by the console surgeon during robot-assisted radical prostatectomy in 10 patients. The transducer is designed as a drop-in probe with a flexible cord for insertion through a laparoscopic port, and is controlled by a robotic arm with the ultrasonographic image shown as a console Tile-pro display. Intraoperative localization of the biopsy-proven cancerous hypoechoic lesion was feasible in four out of four cases. The microtransducer facilitated identification of the bladder neck as well as the appropriate level of neurovascular bundle release. Negative surgical margin was achieved in all 10 cases (100%), even though five of 10 patients (50%) had extraprostatic (pT3) disease. Recovery of erectile function and continence was encouraging. In conclusion, intraoperative ultrasound navigation using a drop-type microtransducer is a novel technique that could enhance the incremental value of the standard information.


Type 3 Secretion System (T3SS) of Bradyrhizobium sp. DOA9 and Its Roles in Legume Symbiosis and Rice Endophytic Association.
The Bradyrhizobium sp. DOA9 strain isolated from a paddy field has the ability to nodulate a wide spectrum of legumes. Unlike other bradyrhizobia, this strain has a symbiotic plasmid harboring nod, nif, and type 3 secretion system (T3SS) genes. This T3SS cluster contains all the genes necessary for the formation of the secretory apparatus and the transcriptional activator (TtsI), which is preceded by a nod-box motif. An in silico search predicted 14 effectors putatively translocated by this T3SS machinery. In this study, we explored the role of the T3SS in the symbiotic performance of DOA9 by evaluating the ability of a T3SS mutant (ΩrhcN) to nodulate legumes belonging to Dalbergioid, Millettioid, and Genistoid tribes. Among the nine species tested, four (Arachis hypogea, Vigna radiata, Crotalaria juncea, and Macroptilium atropurpureum) responded positively to the rhcN mutation (ranging from suppression of plant defense reactions, an increase in the number of nodules and a dramatic improvement in nodule development and infection), one (Stylosanthes hamata) responded negatively (fewer nodules and less nitrogen fixation) and four species (Aeschynomene americana, Aeschynomene afraspera, Indigofera tinctoria, and Desmodium tortuosum) displayed no phenotype. We also tested the role of the T3SS in the ability of the DOA9 strain to endophytically colonize rice roots, but detected no effect of the T3SS mutation, in contrast to what was previously reported in the Bradyrhizobium SUTN9-2 strain. Taken together, these data indicate that DOA9 contains a functional T3SS that interferes with the ability of the strain to interact symbiotically with legumes but not with rice.


Severe injury triggers antigen-specific T-helper cell dysfunction.
Although it is established that post-injury immune dysfunction involves alterations in T-cell function, the effects of injury on T-cell function in vivo are poorly understood. This study uses a mouse injury model and an antigen immunization approach to investigate the influence of injury on antigen-specific T-helper cell function. We report here that injury triggered a significant reduction in antigen-specific T-helper-1 (Th1)-dependent IgG2a antibody formation, while IgM, IgG1, and IgE production was unchanged. In addition, injury caused a reduction in cytokine production (IL-2, IFNgamma and IL-10) by antigen-stimulated T-cells. We also demonstrate that interleukin 12 (IL-12), a cytokine that promotes Th1 cell differentiation, restored IgG2a antibody formation and corrected the injury-induced reduction in antigen-stimulated cytokine production. Taken together, these findings indicate that severe injury induces a dramatic reduction in Th1 cell function in vivo and suggest that therapies designed to restore Th1 cell function may be beneficial to the injured host.


Socioeconomic status and prostate cancer incidence and mortality rates among the diverse population of California.
The racial/ethnic disparities in prostate cancer rates are well documented, with the highest incidence and mortality rates observed among African-Americans followed by non-Hispanic Whites, Hispanics, and Asian/Pacific Islanders. Whether socioeconomic status (SES) can account for these differences in risk has been investigated in previous studies, but with conflicting results. Furthermore, previous studies have focused primarily on the differences between African-Americans and non-Hispanic Whites, and little is known for Hispanics and Asian/Pacific Islanders. To further investigate the relationship between SES and prostate cancer among African-Americans, non-Hispanic Whites, Hispanics, and Asian/Pacific Islanders, we conducted a large population-based cross-sectional study of 98,484 incident prostate cancer cases and 8,997 prostate cancer deaths from California. Data were abstracted from the California Cancer Registry, a population-based surveillance, epidemiology, and end results (SEER) registry. Each prostate cancer case and death was assigned a multidimensional neighborhood-SES index using the 2000 US Census data. SES quintile-specific prostate cancer incidence and mortality rates and rate ratios were estimated using SEER*Stat for each race/ethnicity categorized into 10-year age groups. For prostate cancer incidence, we observed higher levels of SES to be significantly associated with increased risk of disease [SES Q1 vs. Q5: relative risk (RR) = 1.28; 95% confidence interval (CI): 1.25-1.30]. Among younger men (45-64 years), African-Americans had the highest incidence rates followed by non-Hispanic Whites, Hispanics, and Asian/Pacific Islanders for all SES levels. Yet, among older men (75-84 years) Hispanics, following African-Americans, displayed the second highest incidence rates of prostate cancer. For prostate cancer deaths, higher levels of SES were associated with lower mortality rates of prostate cancer deaths (SES Q1 vs. Q5: RR = 0.88; 95% CI: 0.92-0.94). African-Americans had a twofold to fivefold increased risk of prostate cancer deaths in comparison to non-Hispanic Whites across all levels of SES. Our findings suggest that SES alone cannot account for the greater burden of prostate cancer among African-American men. In addition, incidence and mortality rates of prostate cancer display different age and racial/ethnic patterns across gradients of SES.


Modelling the intradermal delivery of microneedle array patches for long-acting antiretrovirals using PBPK.
Existing HIV therapy using oral antiretrovirals (ARVs) can result in pill fatigue and sub-optimal adherence. Microneedle array patches (MAPs) offer non-invasive, blood-free and painless drug delivery, and may improve patient adherence. The objective of this study was to develop a novel physiologically-based pharmacokinetic (PBPK) model to simulate the systemic pharmacokinetics of cabotegravir and rilpivirine MAPs using the intradermal route. The developed PBPK models were qualified against observed pharmacokinetic data after intramuscular (IM) and intradermal administration of long-acting nanoformulated rilpivirine to rats, and for IM administration of both drugs to healthy adults. Qualified models were then utilised to estimate suitable MAP characteristics (e.g. nanoformulation dose and release rates) and inform dosing strategies to maintain plasma concentrations above target trough concentrations for the designated dosing interval. PBPK models simulated q4-weekly loading and maintenance doses of 360 mg and 180 mg for long-acting formulated cabotegravir between the release rates of 1 × 10-3-3 × 10-3h-1 and 1 × 10-3-1.5 × 10-3h-1 respectively, for a 70 kg adult. Estimated patch size was 60 cm2 for a 360 mg dose of cabotegravir. For q4-weekly dosing, rilpivirine required a 1080 mg loading dose and a 540 mg maintenance dose with release rates of 1.5 × 10-3-2.5 × 10-3h-1 and 5 × 10-4-1 × 10-3h-1, respectively. Weekly dosing was also evaluated to assess the potential application from a smaller patch size. The ability to self-administer via a patch that is only left in place for a short duration makes longer durations less important than for some other long-acting approaches. Weekly cabotegravir required 60 mg between release rates 7 × 10-3-9 × 10-3h-1 and rilpivirine required 270 mg and 180 mg respectively between release rates of 7 × 10-3-9 × 10-3h-1. This model estimated optimal dose and release rates for cabotegravir and rilpivirine MAPs. Our approach provides a computational platform to support rational development of intradermal administration strategies to tackle problems associated with chronic oral ARV administration.


Nicotine increased VEGF and MMP2 levels in the rat eye and kidney.
Chronic cigarette smoking affects many tissues negatively. Nicotine in tobacco has negative effects on tissues, kidneys, and eyes especially, where microcirculation is vitally important for the survival and functioning. It is known that appropriate vascular endothelial growth factor (VEGF) and (matrix metalloproteinase 2) MMP2 levels are required for suitable vascularity and enough microcirculation. The aim of this study was to investigate the effect of nicotine on VEGF and MMP2 levels in kidney and eyes, where microcirculation is very important for their function. The nicotine was given into drinking water, to male and female rats for 6 weeks. During the first 2 weeks, the nicotine concentration was 10 mg/L, then was given at a fixed dose of 20 mg/L until the end of the experiment. The VEGF and MMP2 levels were increased in kidney tissue of both genders as a result of given nicotine. MMP2 levels were also increased in the eye tissue for both genders similarly. However, VEGF levels increased in the eye tissue with nicotine in males, whereas it did not change in females. The use of nicotine made VEGF and MMP2 levels increase in kidney tissue in both genders of rats. This increase in VEGF was observed only in male eye tissue, not in females. According to our findings, it can be suggested that nicotine has negative effects on microvascular circulation by increasing VEGF and MMP2 levels. In addition, it should be pointed out that estrogen might have protective effects on female eye tissue. Further studies are necessary to understand the complex relationship between the role of nicotine and estrogen on eye and kidney tissues.


Exploiting cytokine secretion to rapidly produce multivirus-specific T cells for adoptive immunotherapy.
Viral infections remain a major cause of morbidity and mortality after hematopoietic stem cell transplantation (HSCT), and conventional small-molecule therapeutics often have modest benefit, high cost, and adverse effects. Adoptive transfer of donor-derived virus-specific T cells has proved feasible and safe after HSCT and to reconstitute immunity against cytomegalovirus, Epstein-Barr virus, and adenovirus. Current protocols to generate these cytotoxic T cell lines are lengthy, taking up to 12 weeks. As viral infections often occur <30 days after HSCT, speedy production of virus-specific cytotoxic T cells lacking alloreactivity is highly desirable. We now describe a modified rapid selection method for production and characterization of CD4 and CD8 T cells specific for cytomegalovirus, Epstein-Barr virus, and adenovirus in a single infusate. We use Ad5f35-pp65/latent membrane protein 2 vectors in a single procedure over a 48-hour time period and manufacture a product suited for clinical use. By simultaneously expanding a portion of the selected product, we can characterize phenotype and function of the infused product and link them with subsequent in vivo outcome.


Broadband near-field interference spectroscopy of metal nanoparticles using a femtosecond white-light continuum.
We demonstrate a new nanoscale spectroscopic technique that combines subwavelength near-field imaging with broadband interference spectroscopy. We apply this technique to study phase spectra of surface plasmons in individual gold nanoparticles and nanoparticle dimers. Collective plasmon oscillations in selected nanostructures are excited by a femtosecond white-light continuum transmitted through a subwavelength aperture. The interference spectra detected in the far field result from the coherent superposition of the aperture field and the secondary field re-emitted by the nanostructure. The analysis of these spectra allows us to accurately measure the positions and damping constants of single-nanostructure plasmon resonances.


IgG from patients with Guillain-Barré syndrome interact with nicotinic acetylcholine receptor channels.
In Guillain-Barré syndrome (GBS), immunoglobulin G (IgG) antibodies block neuromuscular transmission pre- and postsynaptically and thus are of potential pathogenic relevance. We investigated whether IgG from GBS patients has a direct interaction with nicotinic acetylcholine receptor (nAChR) channels. Purified IgG fractions from six GBS patients that blocked neuromuscular transmission in a previous study were analyzed by the patch-clamp technique in combination with an ultrafast system for solution exchange. Sera from three patients with other inflammatory neurological disorders were used as controls. Mouse myotubes expressing native embryonic-type nAChR channels and human embryonic kidney (HEK) 293 cells transiently transfected with recombinant adult-type nAChR channels were used. Repeated 20-ms pulses of acetylcholine (ACh) were applied to outside-out patches in the presence of GBS-IgG. IgG of the patients had a significant reversible blocking action on embryonic- and adult-type nAChR channels with some variability in the magnitude of the block. Activation and desensitization kinetics were not affected when GBS-IgG was applied. None of the control sera blocked the AChR channels. The observed postsynaptic block effect fulfills the criteria of a channel-blocking IgG antibody similar to those seen in autoimmune myasthenia and may contribute to muscle weakness during the acute phase of GBS.


[Prophylaxis of hearing loss and vision impairment in newborns].
The survival rate of low and very low birth weight infants increases as a natural consequence of the improvement in perinatal care and introducing the new methods and techniques in neonatal intensive therapy. In the face of this progress the number of patients potentially threatened with hearing and vision impairment grows. In this paper we described the most essential risk factors of the occurrence of these complications especially in premature newborns. Also the methods of prevention, early detection of disturbances or treatments of hearing and vision impairment constituted the target of this work.


The opening of the inositol 1,4,5-trisphosphate-sensitive Ca2+ channel in rat cerebellum is inhibited by caffeine.
Ins(1,4,5)P3(InsP3)-induced Ca2+ release and [3H]InsP3 binding were measured in rat cerebellar microsomes in the presence or absence of caffeine. The quantal Ca2+ release was shown to occur in an apparently co-operative fashion with a Hill coefficient (h) of 2.2. Half-maximal Ca2+ release was observed at 900 nM-InsP3. Addition of caffeine caused changes both to the concentration of InsP3 required to cause half-maximal Ca2+ release (3.9 microM at 50 mM-caffeine) and to the apparent co-operativity (h = 1.0 at 50 mM-caffeine). Under standard conditions for [3H]InsP3 binding, caffeine had no effect, and it had no effect on InsP3 metabolism. Cyclic AMP also had no effect on the quantal release induced by InsP3. These results are consistent with the view that caffeine affects the opening (Ca2+ release) events rather than the ligand-binding events in the operation of the InsP3-sensitive Ca2+ channel.


Increases in systemic and local stress: a probable mechanism of visceral fat accumulation and insulin resistance in adult catch-up growth rats?
Catch-up growth in adult (CUGA) is increasingly proposed as an important causative factor for the widespread insulin resistance (IR)-related diseases especially in developing countries/territories. We aimed to investigate the effects of CUGA to insulin sensitivity, lipid profile and stress in rats, as well as the probable relationship among them. Male Sprague-Dawley rats were randomly divided into six groups for two sampling points: caloric restriction group (R4) and normal chow controls for four weeks (NC4); CUGA re-fed with normal chow (RN4), CUGA re-fed with high-fat diet (RH4), normal chow controls (NC8) and high-fat diet controls (HF8) for eight weeks. Visceral fat accumulation (visceral adipose tissue [VAT] percentage), systemic (plasma corticosterone) and local (HSD11B1 mRNA expression in skeletal muscle [SkM] and VAT) stress, whole-body and peripheral insulin sensitivity were determined in this study. After four weeks of caloric restriction, R4 rats showed increases in systemic and local stress, decreases in visceral fat accumulation and no IR (whole-body or peripheral). Yet, after re-feeding, sustained systemic and local stress, remarkable visceral fat accumulation and IR (whole-body and peripheral) were found in RN4 compared with NC8, in RH4 compared with NC8 and HF8. Our findings demonstrated that CUGA rats were characterized by significant IR, visceral fat accumulation and stress. These changes were more severe in CUGA re-fed with high-fat diet. The interaction of sustained caloric restriction-induced stress and re-feeding might be of utmost importance in the etiology of visceral fat accumulation and IR in CUGA.


Safety and efficacy of fluconazole treatment for Candida oesophagitis in AIDS.
The efficacy and safety of fluconazole in the treatment of oesophageal candidiasis in patients with the acquired immunodeficiency syndrome (AIDS) was assessed in 36 patients. Fluconazole, 200 mg orally, was given on the first day, followed by 100 mg daily for 4 weeks. Clinical and mycological evaluation was performed in 31 patients at the end of treatment and 24 were also assessed after 8 weeks of starting treatment. In 1 patient fluconazole was discontinued, 5 patients were lost to follow-up and 6 patients died during the study. Clinical and mycological cure was achieved in all patients; in 31 of 36 patients the clinical picture resolved within a week. The cure was confirmed in 27 patients by oesophagoscopy. Two patients relapsed 1 month after stopping fluconazole but the reinstitution of therapy achieved cure. Asymptomatic fungal oropharynx colonization was evident in about 40% of patients during treatment and follow-up period. Fluconazole was well tolerated by all patients but mild to moderate increase of liver enzymes values occurred in 16. Treatment had to be discontinued in 1 patient with hepatic tuberculosis because of severe liver function abnormalities, but their relation with the drug was uncertain. Fluconazole is an effective and safe treatment of oesophageal candidiasis in AIDS patients.


[Disk-like complex formation by apolipoprotein-lipid membrane interaction and relevance to high-density lipoprotein formation].
The interaction of apolipoprotein A-I (apoA-I) with transmembrane ATP-binding cassette transporter A1 (ABCA1) is a crucial step for high-density lipoprotein (HDL) formation, however, its molecular mechanism is less well understood. Here, we used apoA-I and its model peptide, Ac-18A-NH(2), to investigate their interaction with mixed membranes of phosphatidylcholine (PC) with phosphatidylethanolamine (PE) or sphingomyelin (SM). It was shown that PE, possessing the negative spontaneous curvature, increased both the degree of hydration at the membrane interface and the acyl chain order, and that Ac-18A-NH(2) had opposite effects to PE, since the alpha-helix formation at the membrane surface induced positive curvature strain. In addition, increased PE in PC/PE large unilamellar vesicles (LUVs) enhanced the peptide's binding. Although SM significantly lowered the peptide binding capacity, the peptide's binding to PC/SM LUV led to membrane disruption. The interaction with PC/SM LUVs was then investigated using ApoA-I. The spontaneous rHDL formation from pure PC LUV proceeded very slowly at 37 degrees C, but SM-rich PC/SM LUVs, which are in a gel/liquid-disordered (L(d)) phase at this temperature, were rapidly solubilized to form rHDL by apoA-I. The addition of cholesterol decreased the rate of the rHDL formation and induced the selective extraction of lipids by apoA-I, which preferably extracted lipids of L(d) phase rather than lipids of liquid-ordered (L(o)) phase. These results suggest that heterogeneous interface of the mixed membranes facilitates the insertion of apoA-I and induces L(d) phase-selective lipid extraction to form rHDL, and are compatible with recent cell works on the apoA-I-dependent HDL generation.


Noise analysis and improvement of displacement vector estimation from angular displacements.
Elastography or elasticity imaging techniques typically image local strains or Young's modulus variations along the insonification direction. Recently, techniques that utilize angular displacement estimates obtained from multiple angular insonification of tissue have been reported. Angular displacement estimates obtained along different angular insonification directions have been utilized for spatial-angular compounding to reduce noise artifacts in axial-strain elastograms, and for estimating the axial and lateral components of the displacement vector and the corresponding strain tensors. However, these angular strain estimation techniques were based on the assumption that noise artifacts in the displacement estimates were independent and identically distributed and that the displacement estimates could be modeled using a zero-mean normal probability density function. Independent and identically distributed random variables refer to a collection of variables that have the same probability distribution and are mutually independent. In this article, a modified least-squares approach is presented that does not make any assumption regarding the noise in the angular displacement estimates and incorporates displacement noise artifacts into the strain estimation process using a cross-correlation matrix of the displacement noise artifacts. Two methods for estimating noise artifacts from the displacement images are described. Improvements in the strain tensor (axial and lateral) estimation performance are illustrated utilizing both simulation data obtained using finite-element analysis and experimental data obtained from a tissue-mimicking phantom. Improvements in the strain estimation performance are quantified in terms of the elastographic signal-to-noise and contrast-to-noise ratios obtained with and without the incorporation of the displacement noise artifacts into the least-squares strain estimator.


Transference interpretations, therapeutic alliance, and outcome in short-term individual psychotherapy.
Relationships between the proportion of transference interpretations provided by therapists and both therapeutic alliance and therapy outcome were investigated for a sample of 64 patients who had received approximately 20 sessions of short-term individual psychotherapy within a controlled, clinical trial investigation. Inverse relationships were found between the proportion of transference interpretations and both therapeutic alliance and favorable therapy outcome for patients with a history of high quality of object relations. An examination of individual sessions revealed evidence that was consistent with two different, but not mutually exclusive, causal explanations. The first concerned the negative effects of high proportions of transference interpretations; the second concerned the reaction of the therapist to the presence of a weak therapeutic alliance. While the evidence from our study was correlational, it was sufficiently strong to warrant alerting clinicians to the possibility of negative treatment effects when high levels of transference interpretations are used with certain types of patients receiving short-term individual psychotherapy.


Applications of three-dimensional (3D) printing for microswimmers and bio-hybrid robotics.
This article will focus on recent reports that have applied three-dimensional (3D) printing for designing millimeter to micrometer architecture for robotic motility. The utilization of 3D printing has rapidly grown in applications for medical prosthetics and scaffolds for organs and tissue, but more recently has been implemented for designing mobile robotics. With an increase in the demand for devices to perform in fragile and confined biological environments, it is crucial to develop new miniaturized, biocompatible 3D systems. Fabrication of materials at different scales with different properties makes 3D printing an ideal system for creating frameworks for small-scale robotics. 3D printing has been applied for the design of externally powered, artificial microswimmers and studying their locomotive capabilities in different fluids. Printed materials have also been incorporated with motile cells for bio-hybrid robots capable of functioning by cell contraction and swimming. These 3D devices offer new methods of robotic motility for biomedical applications requiring miniature structures. Traditional 3D printing methods, where a structure is fabricated in an additive process from a digital design, and non-traditional 3D printing methods, such as lithography and molding, will be discussed.


Bedside ultrasonography (US), Echoscopy and US point of care as a new kind of stethoscope for Internal Medicine Departments: the training program of the Italian Internal Medicine Society (SIMI).
In recent years, thanks to the development of miniaturized ultrasound devices, comparable to personal computers, tablets and even to smart phones, we have seen an increasing use of bedside ultrasound in internal medicine departments as a novel kind of ultrasound stethoscope. The clinical ultrasound-assisted approach has proved to be particularly useful in assessing patients with nodules of the neck, dyspnoea, abdominal pain, and with limb edema. In several cases, it has allowed a simple, rapid and precise diagnosis. Since 2005, the Italian Society of Internal Medicine and its Ultrasound Study Group has been holding a Summer School and training courses in ultrasound for residents in internal medicine. A national network of schools in bedside ultrasound was then organized for internal medicine specialists who want to learn this technique. Because bedside ultrasound is a user-dependent diagnostic method, it is important to define the limits and advantages of different new ultrasound devices, to classify them (i.e. Echoscopy and Point of Care Ultrasound), to establish appropriate different levels of competence and to ensure their specific training. In this review, we describe the point of view of the Italian Internal Medicine Society on these topics.


Management of the incidental renal mass.
Despite substantial advances in the imaging-based diagnosis of renal masses, the increased detection of incidental renal masses with cross-sectional imaging poses problems to the radiologist and referring physician. Most incidental renal masses can be diagnosed with confidence and either ignored or treated without further testing. However, some renal masses, particularly small ones, remain indeterminate and require a management strategy that is both medically appropriate and practical. In this article, the literature will be reviewed and an approach to the diagnosis and management of the incidental renal mass will be suggested. Management recommendations, derived from data regarding the probability of malignancy in cystic and solid renal masses, are provided for two types of patients, those in the general population and those with limited life expectancy or co-morbidity. The Bosniak classification is used to guide the management of cystic masses, with observation reserved for selected patients, and the presumption of benignity recommended for simple-appearing cystic masses smaller than 1 cm. Among solid renal masses, a more aggressive overall approach is taken. However, additional imaging, and in selected patients, percutaneous biopsy, is recommended to diagnose benign neoplasms. Although additional studies are needed to establish risks and benefits, observation of solid masses may be considered in selected patients. Minimally invasive treatments of renal cancer (including percutaneous ablation) show promise but at the same time challenge the radiologist to review the approach to the incidental renal mass.


USP1 deubiquitinase maintains phosphorylated CHK1 by limiting its DDB1-dependent degradation.
The maintenance of genetic stability depends on the fine-tuned initiation and termination of pathways involved in cell cycle checkpoints and DNA repair. Here, we describe a new pathway that regulates checkpoint kinase 1 (CHK1) activity, a key element controlling both checkpoints and DNA repair. We show that the ubiquitin-specific peptidase 1 (USP1) deubiquitinase participates in the maintenance of both total and phosphorylated levels of CHK1 in response to genotoxic stress. We establish that USP1 depletion stimulates the damage-specific DNA-binding protein 1-dependent degradation of phosphorylated CHK1 in both a monoubiquitinylated Fanconi anaemia, complementation group D2 (FANCD2)-dependent and -independent manner. Our data support the existence of a circuit in which CHK1 activates checkpoints, DNA repair and proliferating cell nuclear antigen and FANCD2 monoubiquitinylation. The latter two events, in turn, switch off activated CHK1 by negative feedback inhibition, which contributes to the downregulation of the DNA damage response. This pathway, which is compromised in the cancer-prone disease Fanconi anaemia (FA), likely contributes to the hypersensitivity of cells from FA patients to DNA damage and to the clinical phenotype of the syndrome; it may also represent a pharmacological target to improve patient care and develop new cancer therapies.


Coexistence of phage and bacteria on the boundary of self-organized refuges.
Bacteriophage are voracious predators of bacteria and a major determinant in shaping bacterial life strategies. Many phage species are virulent, meaning that infection leads to certain death of the host and immediate release of a large batch of phage progeny. Despite this apparent voraciousness, bacteria have stably coexisted with virulent phages for eons. Here, using individual-based stochastic spatial models, we study the conditions for achieving coexistence on the edge between two habitats, one of which is a bacterial refuge with conditions hostile to phage whereas the other is phage friendly. We show how bacterial density-dependent, or quorum-sensing, mechanisms such as the formation of biofilm can produce such refuges and edges in a self-organized manner. Coexistence on these edges exhibits the following properties, all of which are observed in real phage-bacteria ecosystems but difficult to achieve together in nonspatial ecosystem models: (i) highly efficient virulent phage with relatively long lifetimes, high infection rates and large burst sizes; (ii) large, stable, and high-density populations of phage and bacteria; (iii) a fast turnover of both phage and bacteria; and (iv) stability over evolutionary timescales despite imbalances in the rates of phage vs. bacterial evolution.


Variational denoising of partly textured images by spatially varying constraints.
Denoising algorithms based on gradient dependent regularizers, such as nonlinear diffusion processes and total variation denoising, modify images towards piecewise constant functions. Although edge sharpness and location is well preserved, important information, encoded in image features like textures or certain details, is often compromised in the process of denoising. We propose a mechanism that better preserves fine scale features in such denoising processes. A basic pyramidal structure-texture decomposition of images is presented and analyzed. A first level of this pyramid is used to isolate the noise and the relevant texture components in order to compute spatially varying constraints based on local variance measures. A variational formulation with a spatially varying fidelity term controls the extent of denoising over image regions. Our results show visual improvement as well as an increase in the signal-to-noise ratio over scalar fidelity term processes. This type of processing can be used for a variety of tasks in partial differential equation-based image processing and computer vision, and is stable and meaningful from a mathematical viewpoint.


Dynamic heat capacity and relaxation time of ultraviscous melt and glassy acetaminophen.
The real and imaginary components, C'(p) and C''(p), of the complex heat capacity, C*(p)=C'(p)-iC"(p) of supercooled, ultraviscous melt of acetaminophen have been measured at different temperatures during cooling through its vitrification range and during heating through its glass-softening range by using a modulation frequency of 3.3 mHz. From these data, the distribution of relaxation time parameter, beta, and a characteristic (calorimetric or configurational) relaxation time, tau(cal), have been determined. A constant value of 0.65 for beta fits the data, and tau(cal) varies with the temperature according to the Vogel-Fulcher-Tammann equation, tau(cal) = 10(-12.95) exp[1813/(T - 240.5)]. This relation differs significantly from the one deduced by others in which the configurational entropy theory was used to deduce tau(cal). The C'(p) and C''(p) values measured during the cooling of its ultraviscous melt and during the heating of its glassy state show a small hysteresis only at low temperatures. These investigations also provide a comparison of calorimetric and dielectric relaxation times in ultraviscous acetaminophen and highlight the role of faster modes of relaxation at low temperatures in organic, molecular glasses that can help in a better understanding of the crystal nucleation process in glasses at T below their T(g).


An extended loop in CE7 carbohydrate esterase family is dispensable for oligomerization but required for activity and thermostability.
The carbohydrate esterase family 7 (CE7) belonging to the α/β hydrolase superfamily contains a structurally conserved loop extension element relative to the canonical α/β hydrolase fold. This element called the β-interface loop contributes 20-30% of the total buried surface area at intersubunit interfaces of the functional hexameric state. To test whether this loop is an enabling region for the structure and function of the oligomeric assembly, we designed a truncation variant of the thermostable CE7 acetyl esterase from Thermotoga maritima (TmAcE). Although deletion of 26 out of 40 residues in the loop had little impact on the hexamer formation, the variant exhibited altered dynamics of the oligomeric assembly and a loss of thermal stability. Furthermore, the mutant lacked catalytic activity. Crystal structures of the variant and a new crystal form of the wild type protein determined at 2.75Å and 1.76Å, respectively, provide a rationale for the properties of the variant. The hexameric assembly in the variant is identical to that of the wild type and differed only in the lack of buried surface area interactions at the original intersubunit interfaces. This is accompanied by disorder in an extended region of the truncated loop that consequently induces disorder in the neighboring oxyanion hole loop. Overall, the results suggest that the β-interface loop in CE7 enzymes is dispensable for the oligomeric assembly. Rather, the loop extension event was evolutionarily selected to regulate activity, conformational flexibility and thermal stability.


Accuracy of indirect bracket placement with medium-soft, transparent, broad-coverage transfer trays fabricated using computer-aided design and manufacturing: An in-vivo study.
The objective of this study was to test the precision of in-vivo indirect bracket placement via medium-soft, transparent, broad-coverage, computer-aided designed and manufactured transfer trays using an automated digital method. Seventeen patients requiring vestibular fixed appliances were consecutively recruited, and bonding accuracy was measured at each bracket, evaluating 3 linear (mesiodistal, buccolingual, and vertical) and 3 angular measurements (torque, tip, and rotation) with an automated method involving digital superimposition of individual teeth. Mean and standard deviation values were calculated for both arches, single arch, and tooth type, and the percentages of single deviations over the thresholds of 0.25 mm and 1° were calculated, as well as maximum and minimum values for each deviation and directional bias. Correlations between each variable (arch, tooth type, and single tooth) and deviations were investigated through classification and regression trees (CART) predictive models. Neither mean nor single linear deviations ever exceeded the set cutoff value of 0.25 mm. Mean angular deviations never exceeded 1°, but some individual angular deviations did, specifically 8.31% of torque, 13.16% of tip, and 7.16% of rotation deviations. The highest percentage of deviation was recorded for rotation of the maxillary incisors (18.11%). No evident trend in directional deviation bias was found. Tooth type appears to influence mesiodistal and torque deviations, whereas the single tooth variable influenced the percentage of rotation deviations exceeding 1° (P <0.05). This computer-aided designed and manufactured medium-soft, transparent transfer tray provides accurate bracket placement and could be recommended for routine fixed appliance treatment.


Effect of Preoperative Home-Based Exercise Training on Quality of Life After Lung Cancer Surgery: A Multicenter Randomized Controlled Trial.
Preoperative exercise training is recommended for improvement of clinical outcomes after lung cancer (LC) surgery. However, its effectiveness in preventing postoperative decline in quality of life (QoL) remains unknown. This study investigated the effect of preoperative home-based exercise training (PHET) on QoL after LC surgery. Patients awaiting LC resection were randomized to PHET or a control group (CG). The PHET program combined aerobic and resistance exercise, with weekly telephone supervision. Primary outcome was QoL-assessed with the European Organization for Research and Treatment of Cancer (EORTC) Quality of Life Questionnaire C30 (QLQ-C30) at baseline, before surgery, and 1 month after surgery. The secondary outcomes were hospital length of stay and physical performance. The main analysis included a factorial repeated-measures analysis of variance. Additionally, the proportion of patients experiencing clinical deterioration from baseline to post-surgery was assessed. The study included 41 patients (68.1 ± 9.3 years; 68.3% male) in the intention-to-treat analysis (20 PHET patients, 21 CG patients). A significant group × time interaction was observed for global QoL (p = 0.004). Between-group differences in global QoL were statistically and clinically significant before surgery (mean difference [MD], 13.5 points; 95% confidence interval [CI], 2.4-24.6; p = 0.019) and after surgery (MD, 12.4 points; 95% CI, 1.3-23.4; p = 0.029), favoring PHET. Clinical deterioration of global QoL was reported by 71.4% of the CG patients compared with 30 % of the PHET patients (p = 0.003). Between-group differences in favor of PHET were found in pain and appetite loss as well as in physical, emotional and role functions after surgery (p < 0.05). Compared with CG, PHET was superior in improving preoperative five-times sit-to-stand and postoperative exercise capacity (p < 0.05). No between-group differences in other secondary outcomes were observed. The study showed that PHET can effectively prevent the decline in QoL after LC surgery.


Searching for a schizophrenia susceptibility gene in the 22q11 region.
To investigate a genetic association for schizophrenia within chromosome 22q11 in a Chinese Han population. The PCR-based restriction fragment length polymorphism (PCR-RFLP) analysis was used to detect three single nucleotide polymorphisms (SNPs), rs165655 (A/G base change) and rs165815 (C/T base change) present in the ARVCF (armadillo repeat gene deletion in velocardiofacial syndrome) locus, and rs756656 (A/C base change) in the LOC128979 (expressed sequence tags, EST) locus, among 100 Chinese family trios consisting of fathers, mothers and affected offspring with schizophrenia. Genotype data were analyzed by using linkage disequilibrium (LD) methods including haplotype relative risk (HRR) analysis, transmission disequilibrium test (TDT) and haplotype transmission analysis. The genotype frequency distributions of three SNPs were all in Hardy-Weinberg equilibrium (P>0.05). Both the HRR and the TDT analysis showed that rs165815 was associated with schizophrenia (chi2=6.447, df=1, P=0.011 and chi2=6.313, df=1, P=0.012, respectively), whereas the other two SNPs did not show any allelic association. The haplotype transmission analysis showed a biased transmission for the rs165655-rs165815 haplotype system (chi2=17.224, df=3, P=0.0006) and for the rs756656-rs165655-rs165815 hapoltype system (chi2=20.965, df=7, P=0.0038). Either the ARVCF gene itself or a nearby locus may confer susceptibility to schizophrenia in a Chinese Han population.


Novel Metrics in the Longitudinal Evaluation of Pain Data in Sickle Cell Disease.
Available modalities for the longitudinal capture and analysis of pain intensity in patients with sickle cell disease (SCD) limit our ability to study intraindividual and interindividual variation in pain and the factors influencing the transition from acute to chronic pain in patients with SCD. The objectives of this study were to determine the feasibility of electronic capture of longitudinal outpatient pain intensity data and to test the applicability of novel metrics in the study of intraindividual and interindividual variation in pain in patients with SCD. Twenty SCD patients aged 13 to 21 submitted 2045 diary days of pain intensity data over 229 days using a web-based electronic pain diary or through text message. Participants reported pain (11-point Numerical Rating Score >0) on 1559 diary days (76.2%) suggesting a significant outpatient pain burden. In addition to mean maximum daily pain (MMDP), using maximum daily pain (MDP) scores, we calculated the ninetieth percentile (p90) of MDP, proportion of pain-free days (PPFD), Standard Deviation (SD) of MDP and coefficient of variation (CV) of MDP. Although p50 of MDP and p90 of MDP correlated positively with MMDP, PPFD correlated negatively with MMDP and both MMDP and PPFD correlated poorly with the SD of MDP. Examination of graphic representation of pain trends demonstrated how patients with similar MMDP had varying p90, PPFD, SD/coefficient of variation, and ultimately burden of pain over time. Missing data rates were lowest in the first 30 days of reporting and increased over time. Study participants reported a positive experience with momentary pain reporting and improved communication with health care providers regarding pain. The longitudinal collection of pain data with the inclusion of hospital data during periods of hospitalization is feasible and acceptable in patients with SCD over periods of 30 to 60 days. Long-term collection of pain diary data, while informative, is associated with higher rates of missing data. Novel metrics of pain have the potential to better describe intraindividual and interindividual variation in pain, inform studies of the transition from acute to chronic pain as well as contribute patient-reported end points of pain for interventional clinical trials of pain in SCD.


Biomolecular condensates in fungi are tuned to function at specific temperatures.
Temperature can impact every reaction and molecular interaction essential to a cell. For organisms that cannot regulate their own temperature, a major challenge is how to adapt to temperatures that fluctuate unpredictability and on variable timescales. Biomolecular condensation offers a possible mechanism for encoding temperature-responsiveness and robustness into cell biochemistry and organization. To explore this idea, we examined temperature adaptation in a filamentous-growing fungus called Ashbya gossypii that engages biomolecular condensates containing the RNA-binding protein Whi3 to regulate mitosis and morphogenesis. We collected wild isolates of Ashbya that originate in different climates and found that mitotic asynchrony and polarized growth, which are known to be controlled by the condensation of Whi3, are temperature sensitive. Sequence analysis in the wild strains revealed changes to specific domains within Whi3 known to be important in condensate formation. Using an in vitro condensate reconstitution assay we found that temperature impacts the relative abundance of protein to RNA within condensates and that this directly impacts the material properties of the droplets. Finally, we found that exchanging Whi3 genes between warm and cold isolates was sufficient to rescue some, but not all, condensate-related phenotypes. Together these data demonstrate that material properties of Whi3 condensates are temperature sensitive, that these properties are important for function, and that sequence optimizes properties for a given climate.


Use of intuition by nursing students: instrument development and testing.
Intuition has been described as an important type of nursing knowledge and has gained acceptance as a valid way of knowing in clinical nursing. Use of intuition has become one way of explaining professional expertise. Measures of the use of intuition have been developed primarily for experienced nurses, but few measures of intuition use among nursing students exist. The aim of this paper is to describe the development and psychometric testing of an instrument to measure use of intuition by nursing students. Instrument development consisted of concept clarification, item development, and psychometric testing. Intuition was defined as a non-linear process of knowing perceived through physical awareness, emotional awareness, and/or through physical or spiritual connections. Initial measurement items were derived inductively from the literature and informal interviews with senior nursing students. Seven content experts established a content validity index of 0.86 for the initial 33-item measure. It was pretested with a sample of 20 senior nursing students prior to its nationwide validation with a random sample of 1000 senior bachelor of science and associate degree nursing students. Postal mail data collection resulted in 349 responses (35% response rate). Principal component analysis with orthogonal varimax rotation resulted in seven factors accounting for 66.2% of the variance: physical sensations (28.4%); premonitions (9.7%); spiritual connections (7.7%); reading of cues (6.1%); sensing energy (5.7%); apprehension (4.3%); and reassuring feelings (4%). Eigenvalues ranged from 1 to 7.1 and factor loadings ranged from 0.534 to 0.858. The validation resulted in a revised 25-item measure that demonstrated an overall Cronbach's alpha of 0.89 and a range of 0.69-0.84 for each factor. The study is limited by the use of a self-report measure and the attrition in the randomized sample. The intuition measure for use with students showed evidence of construct validity and reliability. With further testing, the measure could serve as a stimulus to foster students' intuitive abilities.


Protein kinases in human leukemic cells.
Protein kinase activities and cyclic AMP binding capacity were investigated in human peripheral blood cells from leukemic patients and normal controls. Using [gamma 32P] ATP as phosphoryldonor, the phosphorylating activities were not found to be significantly different in either normal or leukemic cells when measured on both artificial basic and acidic substrates. In contrast, the GTP-dependent casein kinase activity, CK2, which is almost undetectable in normal granulocytes, was markedly increased in highly proliferating myeloblastic cells from patients with acute myelogenous leukemia (AML) or with chronic myelogenous leukemia in blastic crisis (BC-CML). Levels of endogenous phosphotyrosine were not higher in leukemic cells than in normal peripheral lymphocytes or granulocytes. Finally, cAMP binding capacity was found to be increased in several types of proliferating leukemic cells, due to a higher amount of the R1-type regulatory subunit of the cAMP-dependent protein kinases. Specific patterns of cAMP binding proteins observed in the different types of normal blood cells were rather blurred in leukemic cells. In conclusion, modifications observed in human leukemic cells seem to be more related to proliferation or blockage in normal differentiation than to their cellular origin.


A wireless, skin-interfaced biosensor for cerebral hemodynamic monitoring in pediatric care.
The standard of clinical care in many pediatric and neonatal neurocritical care units involves continuous monitoring of cerebral hemodynamics using hard-wired devices that physically adhere to the skin and connect to base stations that commonly mount on an adjacent wall or stand. Risks of iatrogenic skin injuries associated with adhesives that bond such systems to the skin and entanglements of the patients and/or the healthcare professionals with the wires can impede clinical procedures and natural movements that are critical to the care, development, and recovery of pediatric patients. This paper presents a wireless, miniaturized, and mechanically soft, flexible device that supports measurements quantitatively comparable to existing clinical standards. The system features a multiphotodiode array and pair of light-emitting diodes for simultaneous monitoring of systemic and cerebral hemodynamics, with ability to measure cerebral oxygenation, heart rate, peripheral oxygenation, and potentially cerebral pulse pressure and vascular tone, through the utilization of multiwavelength reflectance-mode photoplethysmography and functional near-infrared spectroscopy. Monte Carlo optical simulations define the tissue-probing depths for source-detector distances and operating wavelengths of these systems using magnetic resonance images of the head of a representative pediatric patient to define the relevant geometries. Clinical studies on pediatric subjects with and without congenital central hypoventilation syndrome validate the feasibility for using this system in operating hospitals and define its advantages relative to established technologies. This platform has the potential to substantially enhance the quality of pediatric care across a wide range of conditions and use scenarios, not only in advanced hospital settings but also in clinics of lower- and middle-income countries.


Overcoming the heat-sink phenomenon: successful radiofrequency thermal ablation of liver tumors in contact with blood vessels.
To evaluate the efficacy and safety of computed tomography (CT)-guided radiofrequency thermal ablation (RFA) of liver tumors (hepatocellular carcinoma and liver metastases) > 3 mm in diameter that were in contact with blood vessels. During a 3-year period RFA was performed in 28 patients (age range, 36-83 years; male/female ratio, 17:11) with liver tumors (primary and metastatic) that were in contact with blood vessels > 3 mm in diameter. Tumor diameter ranged from 1.7 to 5.1 cm. To evaluate the immediate response, dual-phase dynamic CT images were obtained after intravenous contrast material administration. Imaging follow-up was at 1, 3, 6, and 12 months post-RFA, and every year thereafter. All of 28 patients were treated with a total of 36 sessions. In 22 (79%) of the patients, complete ablation of the tumor was achieved. The remaining 6 (21%) patients showed irregular peripheral enhancement and underwent a second session. At 1-year follow-up 2 of the tumors showed a recurrent lesion and a new ablation was performed. The local tumor progression rate at 1-year follow-up was 8.7% and disease-free survival was achieved in 82.1% of the patients. Complications occurred in 4 patients (14.3%); 2 patients presented with a small sub-capsular hematoma, and 2 patients had a partial liver infarction. RFA is a safe and effective method, even with high-risk tumors adjacent to large blood vessels, which can lead to good results with minimal complications and a low rate of tumor progression.


Cytogenetic studies of pesticide and herbicide sprayers.
Fifty-seven herbicide and pesticide sprayers were studied for the incidence of sister chromatid exchanges (SCEs) in peripheral blood lymphocyte chromosomes. Overall there was no difference between the control group and the sprayers. Five sprayers had an SCE rate three standard deviations outside the mean of the control group. Possible factors contributing to this elevated SCE rate were (1) the failure to use protective clothing and (2) the length of time spraying. Many sprayers failed to take adequate protective measures when mixing and spraying and chemicals were often improperly stored. It is recommended that all persons using herbicides and pesticides take adequate protective measures so that the risk of fatal poisoning and possible long-term genetic hazard be reduced.


Clinical and economic benefits associated with the achievement of both HbA1c and LDL cholesterol goals in veterans with type 2 diabetes.
This study compared the clinical and economic benefits associated with dual-goal achievement, glycated hemoglobin (HbA1c)<7% (53 mmol/mol) and LDL cholesterol (LDL-C)<100 mg/dL, with achievement of only the LDL-C goal or only the HbA1c goal in veterans with type 2 diabetes mellitus (T2DM). This retrospective cohort analysis evaluated electronic medical records (Veterans Integrated Service Network 16) in adult T2DM patients with two or more measurements of LDL-C and HbA1c between 1 January 2004 and 30 June 2010 (N=75,646). Cox proportional hazards models were used to compare microvascular and cardiovascular outcomes by goal achievement status; generalized linear regression models were used to assess diabetes-related resource utilization (hospitalization days and number of outpatient visits) and medical service costs. Relative to achievement of only the LDL-C goal, dual-goal achievement was associated with lower risk of microvascular complications (adjusted hazard ratio [aHR] 0.79), acute coronary syndrome (0.88), percutaneous coronary intervention (0.78), and coronary artery bypass graft (CABG) (0.74); it was also associated with fewer hospitalization days (adjusted incidence rate ratio [aIRR] 0.93) and outpatient visits (0.88), as well as lower diabetes-related annual medical costs (-$130.89). Compared with achievement of only the HbA1c goal, dual-goal achievement was associated with lower risk of the composite cardiovascular-related end point (aHR 0.87) and CABG (aHR 0.62), as well as fewer outpatient visits (aIRR 0.98). Achieving both HbA1c and LDL-C goals in diabetes care is associated with additional clinical and economic benefits, as compared with the achievement of either goal alone.


Mechanical properties of the premature lung: From tissue deformation under load to mechanosensitivity of alveolar cells.
Many preterm infants require mechanical ventilation as life-saving therapy. However, ventilation-induced overpressure can result in lung diseases. Considering the lung as a viscoelastic material, positive pressure inside the lung results in increased hydrostatic pressure and tissue compression. To elucidate the effect of positive pressure on lung tissue mechanics and cell behavior, we mimic the effect of overpressure by employing an uniaxial load onto fetal and adult rat lungs with different deformation rates. Additionally, tissue expansion during tidal breathing due to a negative intrathoracic pressure was addressed by uniaxial tension. We found a hyperelastic deformation behavior of fetal tissues under compression and tension with a remarkable strain stiffening. In contrast, adult lungs exhibited a similar response only during compression. Young's moduli were always larger during tension compared to compression, while only during compression a strong deformation-rate dependency was found. In fact, fetal lung tissue under compression showed clear viscoelastic features even for small strains. Thus, we propose that the fetal lung is much more vulnerable during inflation by mechanical ventilation compared to normal inspiration. Electrophysiological experiments with different hydrostatic pressure gradients acting on primary fetal distal lung epithelial cells revealed that the activity of the epithelial sodium channel (ENaC) and the sodium-potassium pump (Na,K-ATPase) dropped during pressures of 30 cmH2O. Thus, pressures used during mechanical ventilation might impair alveolar fluid clearance important for normal lung function.


Internet use frequency and patient-centered care: measuring patient preferences for participation using the health information wants questionnaire.
The Internet is bringing fundamental changes to medical practice through improved access to health information and participation in decision making. However, patient preferences for participation in health care vary greatly. Promoting patient-centered health care requires an understanding of the relationship between Internet use and a broader range of preferences for participation than previously measured. To explore (1) whether there is a significant relationship between Internet use frequency and patients' overall preferences for obtaining health information and decision-making autonomy, and (2) whether the relationships between Internet use frequency and information and decision-making preferences differ with respect to different aspects of health conditions. The Health Information Wants Questionnaire (HIWQ) was administered to gather data about patients' preferences for the (1) amount of information desired about different aspects of a health condition, and (2) level of decision-making autonomy desired across those same aspects. The study sample included 438 individuals: 226 undergraduates (mean age 20; SD 2.15) and 212 community-dwelling older adults (mean age 72; SD 9.00). A significant difference was found between the younger and older age groups' Internet use frequencies, with the younger age group having significantly more frequent Internet use than the older age group (younger age group mean 5.98, SD 0.33; older age group mean 3.50, SD 2.00; t436=17.42, P<.01). Internet use frequency was positively related to the overall preference rating (γ=.15, P<.05), suggesting that frequent Internet users preferred significantly more information and decision making than infrequent Internet users. The relationships between Internet use frequency and different types of preferences varied: compared with infrequent Internet users, frequent Internet users preferred more information but less decision making for diagnosis (γ=.57, P<.01); more information and more decision-making autonomy for laboratory test (γ=.15, P<.05), complementary and alternative medicine (γ=.32, P<.01), and self-care (γ=.15, P<.05); and less information but more decision-making autonomy for the psychosocial (γ=-.51, P<.01) and health care provider (γ=-.27, P<.05) aspects. No significant difference was found between frequent and infrequent Internet users in their preferences for treatment information and decision making. Internet use frequency has a positive relationship with the overall preferences for obtaining health information and decision-making autonomy, but its relationship with different types of preferences varies. These findings have important implications for medical practice.


[Appearance of entodermal antigen in the mouth mucosa of dogs in recurrent spontaneous stomatitis].
An additional component absent in the same areas of the mouth under normal conditions appeared in dogs with spontaneous stomatitis in the ectodermal derivatives (mucosa of the cheeks, tongue and the hard palate). This component proved to be fully identical to the antigens of the mucosa of the soft palate, esophagus, stomach, small and large intestine, which were entodermal derivatives. No entodermal antigen was revealed in the other organs and tissues under study.


Multi-parametric Ultrasound Evaluation of Pediatric Thyroid Dyshormonogenesis.
The aim of this study was to assess the diagnostic contribution of gray-scale ultrasonography, color Doppler, superb microvascular imaging and shear wave elastography in thyroid dyshormonogenesis (TD). From October 2017 to February 2018, the prospective study included 31 patients (13.6 y; 11-14 y) diagnosed with TD based on thyroid scintigraphy and perchlorate discharge tests and 40 healthy pediatric volunteers (12.8 y; 10-16 y). Median resistive indices (RIs), peak systolic and end-diastolic velocities, vascularity indices (VIs) via superb microvascular imaging and shear wave elastography parameters were evaluated. Median VI values were significantly higher and median RI values were significantly lower in the study group than the control group. No significant difference was found between shear wave elastography parameters of the TD and control group. VI was significantly correlated with median total thyroid gland volumes (p = 0.002, r = 0.28), medication dosage (p = 0.03, r = 0.48) and 2-h radioactive iodine uptake values (p = 0.008, r = 0.57). VI is a clinically significant and novel parameter useful for diagnosing TD.


Posttraumatic stress disorder: a model of the longitudinal course and the role of risk factors.
Posttraumatic stress disorder (PTSD) differs from other anxiety disorders in that experience of a traumatic event is necessary for the onset of the disorder. The condition runs a longitudinal course, involving a series of transitional states, with progressive modification occurring with time. Notably, only a small percentage of people that experience trauma will develop PTSD. Risk factors, such as prior trauma, prior psychiatric history, family psychiatric history, peritraumatic dissociation, acute stress symptoms, the nature of the biological response, and autonomic hyperarousal, need to be considered when setting up models to predict the course of the condition. These risk factors influence vulnerability to the onset of PTSD and its spontaneous remission. In the majority of cases, PTSD is accompanied by another condition, such as major depression, an anxiety disorder, or substance abuse. This comorbidity can also complicate the course of the disorder and raises questions about the role of PTSD in other psychiatric conditions. This article reviews what is known about the emergence of PTSD following exposure to a traumatic event using data from clinical studies.


Chlorin e6 and polydopamine modified gold nanoflowers for combined photothermal and photodynamic therapy.
Combinational photo-based approaches with enhanced efficacy for cancer therapy have garnered increasing attention in recent years. In this work, a multifunctional system for synergistic photothermal and photodynamic cancer therapy was successfully prepared. The system consists of gold nanoflowers (AuNFs) conjugated with Chlorin e6 (Ce6), and then coated with a polydopamine (PDA) layer. AuNFs with diameters around 80 nm and a broad absorbance in the visible-near infrared (Vis-NIR) range of 500 to 800 nm, were successfully synthesized by a two-step process at 0 °C, using HAuCl4, ascorbic acid (AA), and hydroxylamine hydrochloride (NH2OH·HCl) as the reactants. Glutathione (GSH) molecules chemically anchored to the gold surfaces were used to provide addressable sites for Ce6 conjugated to GSH-AuNFs through an amidation reaction. A PDA layer was then wrapped outside the Ce6-GSH-AuNFs via self-polymerization of dopamine, which provided additional chemical modification and functionalization. Finally, the multifunctional PDA-Ce6-GSH-AuNFs were obtained. The content of Ce6 incorporated into the AuNFs was 14.0 wt%, and the singlet oxygen yield of PDA-Ce6-GSH-AuNFs was approximately 91.0% of that of free Ce6. PDA-Ce6-GSH-AuNFs showed better photothermal conversion efficiency (η = 23.6%), lower cytotoxicity, and faster cell internalization. Both in vitro and in vivo investigation of the combined treatment with a near-infrared (NIR) laser (660 nm for photodynamic therapy, and 808 nm for photothermal therapy) demonstrated that PDA-Ce6-GSH-AuNFs had excellent phototoxicity and synergistic effects of killing cancer cells. Hence, PDA-Ce6-GSH-AuNFs are a dual phototherapeutic agent that exhibits photodynamic and photothermal therapeutic effects and has potential application in enhanced cancer therapy.


The oncogenic potential of an activated Hox-2.4 homeobox gene in mouse fibroblasts.
The homeobox gene Hox-2.4 is transcriptionally activated in cells of the mouse myeloid leukemia WEHI-3B. The constitutive Hox-2.4 expression in WEHI-3B cells is due to insertion of a transposable element belonging to the family of intracisternal A particles. In this study, we demonstrated the oncogenic potential of this activated homeobox gene. NIH 3T3 fibroblast clones bearing the activated Hox-2.4 gene produced fibrosarcomas in nude mice.


Using non-Gaussian density functional fits to improve relative free energy calculations.
The accurate and reliable computation of relative free energy differences remains an important long-term goal. Major stumbling blocks for achieving this goal reflect the difficulty of sampling in a known fashion along the reaction coordinate and of maximally combining information that has been collected from the simulation along the reaction coordinate. In this paper we examine the utility of a probability density functional type fit to the distribution of work events collected during a nonequilibrium sample along the reaction coordinate. This approach can readily be generalized to equilibrium sampling and has the potential to estimate the quality of a relative free energy estimate as data are being collected. The method may have the greatest utility for nonequilibrium sampling where non-Gaussian work distributions are generally present that are strongly dominated by rare event sampling in the tail region. We believe that the approach can be used to augment the design and the error analysis of relative free energy computations thus improving the ability to reliably and with known accuracy compute a relative free energy.


Biodiesel production in a semiarid environment: a life cycle assessment approach.
While the use of biodiesel appears to be a promising alternative to petroleum fuel, the replacement of fossil fuel by biofuel may not bring about the intended climate cooling because of the increased soil N2O emissions due to N-fertilizer applications. Using a life cycle assessment approach, we assessed the influence of soil nitrous oxide (N2O) emissions on the life cycle global warming potential of the production and combustion of biodiesel from canola oil produced in a semiarid climate. Utilizing locally measured soil N2O emissions, rather than the Intergovernmental Panel on Climate Change (IPCC) default values, decreased greenhouse gas (GHG) emissions from the production and combustion of 1 GJ biodiesel from 63 to 37 carbon dioxide equivalents (CO2-e)/GJ. GHG were 1.1 to 2.1 times lower than those from petroleum or petroleum-based diesel depending on which soil N2O emission factors were included in the analysis. The advantages of utilizing biodiesel rapidly declined when blended with petroleum diesel. Mitigation strategies that decrease emissions from the production and application of N fertilizers may further decrease the life cycle GHG emissions in the production and combustion of biodiesel.


The association between neuronal nitric oxide synthase and neuronal sensitivity in the brain after brain injury.
Injury to the central nervous system is the leading cause of disability in the United States. Neuronal death is one of the causes of disability. Among patients who survive this type of injury, various degrees of recovery in brain function are observed. The molecular basis of functional recovery is poorly understood. Clinical observations and research using experimental injury models have implicated several metabolites in the cascade of events that lead to neuronal degeneration. The levels of intracellular ATP (energy source) and pH are decreased, whereas levels of extracellular glutamate, intracellular calcium ions, and oxidative damage to RNA/DNA, protein, and lipid are increased. These initiating events can be associated with energy failure and mitochondrial dysfunction, resulting in functional or structural brain damage. The injured brain is known to express immediate early genes. Recent studies show that reactive oxygen species (ROS) cause lesions in genes from which mRNA is transcribed as part of the endogenous neuroprotective response. Although degenerating proteins and lipids may contribute to necrosis significantly after severe injury, abnormalities in genetic material, if not repaired, disturb cellular function at every level by affecting replication, transcription, and translation. These lesions include abnormal nucleic acids, known as oxidative lesions of DNA (ODLs) or of RNA (ORLs). In this review, we focus on our current understanding of the various effects of neuronal nitric oxide synthase on the formation of modified bases in DNA and RNA that are induced in the brain after injury, and how ODLs and ORLs affect cell function.


The geometric curvature of the lumbar spine during restricted and unrestricted squats.
The main purpose of this study was to analyze the behavior of the geometric curvature of the lumbar spine during restricted and unrestricted squats, using a novel investigative method. The rationale for our hypothesis is that the lumbar curvature has different patterns at different spine levels depending on the squat technique used. Spine motion was collected via stereo-photogrammetric analysis in nineteen participants (11 males, 8 females). The reconstructed spine points at the upright neutral position and at the deepest position of the squat exercise were projected onto the sagittal plane of the trunk, a polynomial was fitted to the data, and were quantified the two-dimensional geometric curvature at lower, central and higher lumbar levels, besides the inclination of trunk and lumbosacral region, the overall geometric curvature and overall angle of the lumbar spine. The mean values for each variable were analysed with paired t-test (P<0.05). The lumbar presents a flexion from upright neutral posture to deepest point of the movement, but for the lower lumbar the flexion is less intense if the knees travel anteriorly past the toes. The trunk and the lumbosacral region lean forward in both squat techniques and these effects are also reduced in unrestricted squats. The data collected in the study are evidence that during barbell squats the lumbar curvature has different patterns at different spinal levels depending on the exercise technique. The lower lumbar spine appears to be less overloaded during unrestricted squats.


Chinese hamster ovary cell lysosomes rapidly exchange contents.
We have used cell fusion to address the question of whether macromolecules are rapidly exchanged between lysosomes. Donor cell lysosomes were labeled by the long-term internalization of the fluid-phase pinocytic markers, invertase (sucrase), Lucifer Yellow, FITC-conjugated dextran, or Texas red-conjugated dextran. Recipient cells contained lysosomes swollen by long-term internalization of dilute sucrose or marked by an overnight FITC-dextran uptake. Cells were incubated for 1 or 2 h in marker-free media before cell fusion to clear any marker from an endosomal compartment. Recipient cells were infected with vesicular stomatitis virus as a fusogen. Donor and recipient cells were co-cultured for 1 or 2 h and then fused by a brief exposure to pH 5. In all cases, extensive exchange of content between donor and recipient cell lysosomes was observed at 37 degrees C. Incubation of cell syncytia at 17 degrees C blocked lysosome/lysosome exchange, although a "priming" process(es) appeared to occur at 17 degrees C. The kinetics of lysosome/lysosome exchange in fusions between cells containing invertase-positive lysosomes and sucrose-positive lysosomes indicated that lysosome/lysosome exchange was as rapid, if not more rapid, than endosome/lysosome exchange. These experiments suggest that in vivo the lysosome is a rapidly intermixing organellar compartment.


Species and sex-dependent toxicokinetics of 1-bromopropane: the role of hepatic cytochrome P450 oxidation and glutathione (GSH).
1. The objectives of the current studies were to evaluate the factors influencing the toxicokinetics of 1-bromopropane (1-BP) in rodents after intravenous (IV) and inhalation exposure. 2. F-344 rats were administered 1-BP via IV bolus injection at 5 and 20 mg/kg and blood concentration determined versus time. F-344 rats and B6C3F1 mice were also exposed to starting inhalation concentrations 70, 240, 800 and 2700 ppm 1-BP in a closed gas uptake system and chamber 1-BP levels were monitored for 6 h. Plasma bromide concentrations were determined to estimate total metabolized dose. Rats were pretreated with chemical inhibitors of cytochrome P450 and glutathione (GSH) synthesis, prior to exposure to 1-BP at 800 ppm within inhalation chambers. 3. Systemic clearance of 1-BP in rat was rapid and decreased with increasing dose. As inhalation chamber concentration of 1-BP increased, the terminal elimination rates decreased. Half-life of 1-BP in rats following inhibition of P450 (9.6 h) or depletion of GSH (4.1 h) increased relative to controls (2.0 h) at 800 ppm. The percentage of 1-BP metabolized decreased with increasing inhalation exposure. Hepatic levels of GSH were significantly lowered regardless of the exposure level in both rats and mice. Chamber concentration-time curves were fit to a two compartment model which was used to estimate metabolic rate constants. 4. These data suggest that in rat, 1-BP clearance is saturable and that elimination is highly dependent on both P450 and GSH-dependent metabolism. This investigation in rodents may provide an understanding of interspecies differences in toxicokinetics and eventually aid translation of animal studies to human risk assessment.


Out-of-sequence signal 3 as a mechanism for virus-induced immune suppression of CD8 T cell responses.
Virus infections are known to induce a transient state of immune suppression often associated with an inhibition of T cell proliferation in response to mitogen or cognate-antigen stimulation. Recently, virus-induced immune suppression has been linked to responses to type 1 interferon (IFN), a signal 3 cytokine that normally can augment the proliferation and differentiation of T cells exposed to antigen (signal 1) and co-stimulation (signal 2). However, pre-exposure of CD8 T cells to IFN-inducers such as viruses or poly(I∶C) prior to antigen signaling is inhibitory, indicating that the timing of IFN exposure is of essence. We show here that CD8 T cells pretreated with poly(I∶C) down-regulated the IFN receptor, up-regulated suppressor of cytokine signaling 1 (SOCS1), and were refractory to IFNβ-induced signal transducers and activators of transcription (STAT) phosphorylation. When exposed to a viral infection, these CD8 T cells behaved more like 2-signal than 3-signal T cells, showing defects in short lived effector cell differentiation, reduced effector function, delayed cell division, and reduced levels of survival proteins. This suggests that IFN-pretreated CD8 T cells are unable to receive the positive effects that type 1 IFN provides as a signal 3 cytokine when delivered later in the signaling process. This desensitization mechanism may partially explain why vaccines function poorly in virus-infected individuals.


Glucose-stimulated insulin secretion correlates with changes in mitochondrial and cytosolic Ca2+ in aequorin-expressing INS-1 cells.
Nutrient-stimulated insulin secretion is dependent upon the generation of metabolic coupling factors in the mitochondria of the pancreatic B cell. To investigate the role of Ca2+ in mitochondrial function, insulin secretion from INS-1 cells stably expressing the Ca2+-sensitive photoprotein aequorin in the appropriate compartments was correlated with changes in cytosolic calcium ([Ca2+]c) and mitochondrial calcium ([Ca2+]m). Glucose and KCl, which depolarize the cell membrane, as well as the Ca2+-mobilizing agonist, carbachol (CCh), cause substantial increases in [Ca2+]m which are associated with smaller rises in [Ca2+]c. The L-type Ca2+-channel blocker, SR7037, abolished the effects of glucose and KCl while attenuating the CCh response. Glucose-induced increases in [Ca2+]m, [Ca2+]c, and insulin secretion all demonstrate a pronounced initial peak followed by a sustained plateau. All three parameters are increased synergistically when glucose and CCh are combined. Finally, [Ca2+]m, [Ca2+]c, and insulin secretion also display desensitization phenomena following repeated additions of the three stimuli. The high sensitivity of [Ca2+]m to Ca2+ influx and the desensitization-resensitization effects can be explained by a model in which the mitochondria of INS-1 cells are strategically located to sense Ca2+ influx through plasma membrane Ca2+ channels. In conclusion, the correlation of [Ca2+]m and [Ca2+]c with insulin secretion may indicate a fundamental role for Ca2+ in the adaptation of oxidative metabolism to the generation of metabolic coupling factors and the energy requirements of exocytosis.


Simulation of longitudinal exposure data with variance-covariance structures based on mixed models.
Longitudinal data are important in exposure and risk assessments, especially for pollutants with long half-lives in the human body and where chronic exposures to current levels in the environment raise concerns for human health effects. It is usually difficult and expensive to obtain large longitudinal data sets for human exposure studies. This article reports a new simulation method to generate longitudinal data with flexible numbers of subjects and days. Mixed models are used to describe the variance-covariance structures of input longitudinal data. Based on estimated model parameters, simulation data are generated with similar statistical characteristics compared to the input data. Three criteria are used to determine similarity: the overall mean and standard deviation, the variance components percentages, and the average autocorrelation coefficients. Upon the discussion of mixed models, a simulation procedure is produced and numerical results are shown through one human exposure study. Simulations of three sets of exposure data successfully meet above criteria. In particular, simulations can always retain correct weights of inter- and intrasubject variances as in the input data. Autocorrelations are also well followed. Compared with other simulation algorithms, this new method stores more information about the input overall distribution so as to satisfy the above multiple criteria for statistical targets. In addition, it generates values from numerous data sources and simulates continuous observed variables better than current data methods. This new method also provides flexible options in both modeling and simulation procedures according to various user requirements.