A comparison of autorefractor performance.
To compare the agreement between subjective refraction and autorefraction using two commercially available autorefractors. Prospective data were collected for 190 subjects using either the Nidek ARK-700A (Fremont, CA) or the Topcon KR-8000 (Paramus, NJ) and subjective refraction (masked to autorefraction). Refractions were compared in terms of spherical equivalent using Bland-Altman limits of agreement and astigmatic vector difference using median and 95th percentile. The two groups were similar for age, gender, spherical equivalent, and astigmatic power. The differences in spherical equivalent between subjective and autorefraction were significantly different (mean +/- SD; Nidek, -0.03 +/- 0.36 D; Topcon, +0.11 +/- 0.34 D; analysis of variance, F = 7.84; p < 0.01). However, the 95% limits of agreement were similar: Nidek, -0.74 to +0.68 D; Topcon, -0.55 to +0.77 D. The median differences in astigmatic vector difference were also similar: Nidek, 0.27 D and Topcon, 0.25 D. However, the 95th percentile was 0.67 D for Nidek and 1.09 D for Topcon. There was a low frequency of large (>1.00 D) differences in spherical equivalent, 1.1% with each autorefractor. There were five cases with astigmatic vector difference >1 D, all with the Topcon KR-8000 (5.3%). Both autorefractors show excellent agreement with subjective refraction. Despite a statistically significant difference in mean spherical equivalent (0.14 D), near identical limits of agreement (0.10 D difference) suggest clinical equivalence. Conversely, for astigmatism, despite similar median scores, major outliers were more likely with the Topcon, reflected in a 0.42 D larger 95th percentile, which suggests a small advantage for the Nidek for avoiding large astigmatic errors.


Expression of Vibrio salmonicida virulence genes and immune response parameters in experimentally challenged Atlantic salmon (Salmo salar L.).
The Gram-negative bacterium Vibrio salmonicida is the causative agent of cold-water vibriosis (CV), a hemorrhagic septicemia that primarily affects farmed Atlantic salmon (Salmo salar L.). The mechanisms of disease development, host specificity and adaptation, as well as the immunogenic properties of V. salmonicida are largely unknown. Therefore, to gain more knowledge on the pathogenesis of CV, 90 Atlantic salmon parr were injected intraperitoneally with 6 × 10(6) CFU of V. salmonicida LFI1238. Samples from blood and spleen tissue were taken at different time points throughout the challenge for gene expression analysis by two-step reverse transcription (RT) quantitative real-time polymerase chain reaction. Out of a panel of six housekeeping genes, accD, gapA, and 16S rDNA were found to be the most suitable references for expression analysis in Vibrio salmonicida. The bacterial proliferation during challenge was monitored based on the expression of the 16S rRNA encoding gene. Before day 4, the concentrations of V. salmonicida in blood and spleen tissue demonstrated a lag phase. From day 4, the bacterial proliferation was exponential. The expression profiles of eight genes encoding potential virulence factors of V. salmonicida were studied. Surprisingly, all tested virulence genes were generally highest expressed in broth cultures compared to the in vivo samples. We hypothesize that this general muting of gene expression in vivo may be a strategy for V. salmonicida to hide from the host immune system. To further investigate this hypothesis, the expression profiles of eight genes encoding innate immune factors were analyzed. The results demonstrated a strong and rapid, but short-lasting innate immune response against V. salmonicida. These results suggest that the bacterium possesses mechanisms that inhibit and/or resist the salmon innate immune system until the host becomes exhausted of fighting the on-going and eventually overwhelming infection.


Entrapped central venous catheter after mitral valve replacement and its surgical retrieval.
Central venous pressure monitoring line insertion is routine prior to the conduct of cardiac surgery, and in rare instances, malposition can contribute to operative complications. We describe here how a central venous line lying in the right atrium became caught in a left atrial (LA) closure suture during a mitral valve replacement. The opening of the LA suture line is highly unsafe without cardiopulmonary bypass (CPB) because of the possibility of systemic air embolism, but by employing an ingenious method of suturing over and unravelling the continuous sutures closing the left atrium, it was possible to surgically retrieve it without the use of a CPB.


Adrenal surgery before and after the introduction of laparoscopic adrenalectomy.
The widespread introduction of laparoscopic adrenalectomy (LA) may have broadened the indications for a surgical approach to adrenal lesions. The aim of this study was to evaluate how the introduction of the laparoscopic technique changed the profile of patients referred for adrenal surgery. Some 243 patients, who underwent adrenal surgery between November 1986 and December 1999, were divided in two groups: patients in group 1 underwent surgery before the introduction of LA (November 1986 to May 1993) and those in group 2 were operated on after the introduction of LA (June 1993 to December 1999). The groups were compared with respect to gender, age, preoperative diagnosis and lesion size. There were 88 patients in group 1 and 155 in group 2. The mean number of adrenal operations per month was 1.1 during the first period and 2.0 during the second. No differences were found in gender and lesion size. The statistical analysis showed a significant difference in indications for surgery (P = 0.005). The patients in group 2 were older but, after excluding those with metastases, the difference between the two groups was not significant. The introduction of LA resulted in an increase in the number of patients referred. The criteria for patient selection did not change, but more patients with adrenal metastases and incidentaloma were operated on laparoscopically.


A Conserved Amino Acid in the C Terminus of Human Papillomavirus E7 Mediates Binding to PTPN14 and Repression of Epithelial Differentiation.
The human papillomavirus (HPV) E7 oncoprotein is a primary driver of HPV-mediated carcinogenesis. The E7 proteins from diverse HPVs bind to the host cellular nonreceptor protein tyrosine phosphatase type 14 (PTPN14) and direct it for degradation through the activity of the E7-associated host E3 ubiquitin ligase UBR4. Here, we show that a highly conserved arginine residue in the C-terminal domain of diverse HPV E7 mediates the interaction with PTPN14. We found that disruption of PTPN14 binding through mutation of the C-terminal arginine did not impact the ability of several high-risk HPV E7 proteins to bind and degrade the retinoblastoma tumor suppressor or activate E2F target gene expression. HPVs infect human keratinocytes, and we previously reported that both PTPN14 degradation by HPV16 E7 and PTPN14 CRISPR knockout repress keratinocyte differentiation-related genes. Now, we have found that blocking PTPN14 binding through mutation of the conserved C-terminal arginine rendered both HPV16 and HPV18 E7 unable to repress differentiation-related gene expression. We then confirmed that the HPV18 E7 variant that could not bind PTPN14 was also impaired in repressing differentiation when expressed from the complete HPV18 genome. Finally, we found that the ability of HPV18 E7 to extend the life span of primary human keratinocytes required PTPN14 binding. CRISPR/Cas9 knockout of PTPN14 rescued keratinocyte life span extension in the presence of the PTPN14 binding-deficient HPV18 E7 variant. These results support the model that PTPN14 degradation by high-risk HPV E7 leads to repression of differentiation and contributes to its carcinogenic activity.IMPORTANCE The E7 oncoprotein is a primary driver of HPV-mediated carcinogenesis. HPV E7 binds the putative tumor suppressor PTPN14 and targets it for degradation using the ubiquitin ligase UBR4. PTPN14 binds to a C-terminal arginine highly conserved in diverse HPV E7. Our previous efforts to understand how PTPN14 degradation contributes to the carcinogenic activity of high-risk HPV E7 used variants of E7 unable to bind to UBR4. Now, by directly manipulating E7 binding to PTPN14 and using a PTPN14 knockout rescue experiment, we demonstrate that the degradation of PTPN14 is required for high-risk HPV18 E7 to extend keratinocyte life span. Our data show that PTPN14 binding by HPV16 E7 and HPV18 E7 represses keratinocyte differentiation. HPV-positive cancers are frequently poorly differentiated, and the HPV life cycle depends upon keratinocyte differentiation. The finding that PTPN14 binding by HPV E7 impairs differentiation has significant implications for HPV-mediated carcinogenesis and the HPV life cycle.


Sensory reactions of nasal pungency and odor to volatile organic compounds: the alkylbenzenes.
Research assessed the independent contribution of the trigeminal and olfactory nerves to the detection of airborne chemicals by measuring nasal detection thresholds in subjects clinically diagnosed as lacking a functional sense of smell (anosmics) and in matched normal controls (normosmics). Anosmics can provide only odor-unbiased pungency thresholds. Normosmics provided odor thresholds. Stimuli included homologous alkylbenzenes (from toluene to octylbenzene), chlorobenzene, 1-octene, and 1-octyne. As seen before with homologous alcohols, acetates, and ketones, both types of threshold declined with increasing carbon chain length. Anosmics failed to detect alkylbenzenes above propylbenzene. The strong linear correlation between pungency thresholds and saturated vapor concentration for all tested compounds, as a whole, and the constancy of pungency thresholds expressed as percent of vapor saturation, suggests that nasal pungency from these substances relies heavily on a broadly tuned physicochemical interaction with a susceptible biophase within the cell membrane. Through such a nonspecific mechanism, low, subthreshold levels of a wide variety of volatile organic compounds of low reactivity-as found in many polluted indoor spaces-could add their sensory impact to precipitate noticeable sensory irritation.


Cyclooxygenase-2 protein expression modulates cell proliferation and apoptosis in solid ameloblastoma and odontogenic keratocyst. An immunohistochemical study.
Cyclooxygenase-2 protein is a critically important mediator in inflammation that influences proliferation, apoptosis, angiogenesis and metastasis. Previous works showed a relationship between cyclooxygenase-2 and tumourigenesis in humans and animal models. In epithelial odontogenic tumours and cysts, increased cell proliferation and survival have been linked to its pathogenesis and tumour development. The aim of the present study was to analyse the immunohistochemical expression of cyclooxygenase-2 in solid ameloblastoma and odontogenic keratocyst and its association with proteins related to cell proliferation and apoptosis. This study was conducted on 40 cases from the Pathological Anatomy Service, University of Chile. The cases were diagnosed as solid ameloblastoma (n = 21) and odontogenic keratocyst (n = 19) according to WHO 2017. Slides prepared from paraffin-embedded sections were immunohistochemically stained for cyclooxygenase-2, cyclin D1, Ki-67, p63 and Bcl-2. Statistical evaluation was performed by the Shapiro-Wilk test, ANOVA Mann-Whitney test and Spearman's correlation coefficient (p < 0.05). There were significant differences in the immunoexpression of cyclin D1, Ki-67 and Bcl-2 between solid ameloblastoma and odontogenic keratocyst. Likewise, there was a significant difference in the immunoexpression of p63 between follicular and plexiform histological types/subtypes of solid ameloblastoma. Lastly, there were statistical associations between cyclooxygenase-2 and Ki-67 for solid ameloblastoma and between cyclooxygenase-2 and p63 for odontogenic keratocyst. A high level of cyclooxygenase-2 is related to increased cell survival and proliferative activity in solid ameloblastoma and odontogenic keratocyst. This event might contribute to tumoural progression and local invasiveness in these lesions.


Comparative synergistic activity of nafcillin, oxacillin, and methicillin in combination with gentamicin against.
The effectiveness of three semisynthetic, penicillinase-resistant penicillins alone and in combination with gentamicin was tested against 29 clinical isolates of enterococci. The minimal inhibitory concentrations of nafcillin were considerably lower than those of oxacillin and methicillin but were slightly higher than those of penicillin. At clinically achievable concentrations, the combination of nafcillin plus gentamicin produced enhanced killing against 13 of 14 strains of enterococci and was synergistic (by very rigid criteria) against 10 of 14 strains. In contrast, combinations of oxacillin plus gentamicin were synergistic against only 3 of 14 strains, and methicillin plus gentamicin produced synergistic killing against only 1 of 14 strains.


Spatio-Temporal Fluctuations of Neural Dynamics in Mild Cognitive Impairment and Alzheimer's Disease.
An accurate characterization of neural dynamics in mild cognitive impairment (MCI) is of paramount importance to gain further insights into the underlying neural mechanisms in Alzheimer's disease (AD). Nevertheless, there has been relatively little research on brain dynamics in prodromal AD. As a consequence, its neural substrates remain unclear. In the present research, electroencephalographic (EEG) recordings from patients with dementia due to AD, subjects with MCI due to AD and healthy controls (HC) were analyzed using relative power (RP) in conventional EEG frequency bands and a novel parameter useful to explore the spatio-temporal fluctuations of neural dynamics: the spectral flux (SF). Our results suggest that dementia due to AD is associated with a significant slowing of EEG activity and several significant alterations in spectral fluctuations at low (i.e. theta) and high (i.e. beta and gamma) frequency bands compared to HC (p < 0.05). Furthermore, subjects with MCI due to AD exhibited a specific frequency-dependent pattern of spatio-temporal abnormalities, which can help identify neural mechanisms involved in cognitive impairment preceding AD. Classification analyses using linear discriminant analysis with a leave-one-out cross-validation procedure showed that the combination of RP and within-electrode SF at the beta band was useful to obtain a 77.3 % of accuracy to discriminate between HC and AD patients. In the case of comparison between HC and MCI subjects, the classification accuracy reached a value of 79.2 %, combining within-electrode SF at beta and gamma bands. SF has proven to be a useful measure to obtain an original description of brain dynamics at different stages of AD. Consequently, SF may contribute to gain a more comprehensive understanding into neural substrates underlying MCI, as well as to develop potential early AD biomarkers.


Stem cells and rat liver carcinogenesis: contributions of confocal and electron microscopy.
Microscopic analysis in combination with cytochemistry and immunocytochemistry has revealed the presence of four cell types not previously described in the portal area and parenchyma of the liver from an experimental rodent hepatocarcinogenic rat model. Within the intrahepatic bile ductules, which proliferate after administration of chemical carcinogens and partial hepatectomy, small, undifferentiated nonpolarized, nonepithelial cells with a blast-like phenotype and polarized epithelial cells different from the polarized epithelial cells that typically line the walls of the bile ductules were found. In the connective tissue stroma surrounding the bile ductules, nonpolarized epithelial cells with hepatocyte phenotype were found. In the parenchyma, subpopulations of bile ductule epithelial cells that established ATPase-positive bile canalicular structures, including the formation of desmosomes and tight junctions, with parenchymal hepatocytes within the hepatic lobule were found. These observations raise the following questions in this model. Are there undifferentiated progenitor cells with stem cell-like properties within bile ductules? What are the interrelations of the newly described cell types with each other, with parenchymal hepatocytes, with preneoplastic nodules, and with hepatomas? Do the heterogeneous cell types within the bile ductules, in the surrounding connective tissue, and within the hepatic cords represent intermediate stages of single or multiple cell lineage pathways leading to hepatocyte differentiation, liver regeneration, and/or preneoplastic nodule formation?


Physical capacity in women with primary Sjögren's syndrome: a controlled study.
To examine physical capacity (aerobic capacity, joint mobility, muscle function, and standing balance) in women with primary Sjögren's syndrome (primary SS) and to examine the correlation of aerobic capacity with fatigue, functional disability, and mental aspects (anxiety and depression). Fifty-one women who fulfilled the European Community criteria for primary SS and who had anti-SSA/SSB antibodies or a positive lip biopsy were compared with 51 age-matched controls. Physical capacity, fatigue, functional disability, anxiety, and depression were investigated by means of questionnaires and clinical examinations. The women with primary SS had significantly decreased aerobic capacity (VO(2max) = 28.7 versus 32.4 ml/kg/minute; P = 0.013), shoulder mobility (58 versus 59 scale points; P = 0.003), grip strength (214 versus 259 N; P = 0.000), isokinetic strength of the knee flexors (51 versus 56 Nm; P = 0.049), endurance of the knee flexors (620 versus 712 J; P = 0.008), and standing balance (25 versus 28 seconds; P = 0.006) when compared with the reference group. For the primary SS patients, greater effort was needed to carry out the test of aerobic capacity, and they experienced more pain during the shoulder mobility test. Aerobic capacity correlated with the fatigue experienced (r = -0.33, P = 0.022) but not with functional disability or mental aspects. The results indicate that women with primary SS have decreased physical capacity, which may be related to the experience of fatigue.


Evaluation of reverse transcription-PCR assays for rapid diagnosis of severe acute respiratory syndrome associated with a novel coronavirus.
The reverse transcription (RT)-PCR protocols of two World Health Organization (WHO) severe acute respiratory syndrome (SARS) network laboratories (WHO SARS network laboratories at The University of Hong Kong [WHO-HKU] and at the Bernhard-Nocht Institute in Hamburg, Germany [WHO-Hamburg]) were evaluated for rapid diagnosis of a novel coronavirus (CoV) associated with SARS in Hong Kong. A total of 303 clinical specimens were collected from 163 patients suspected to have SARS. The end point of both WHO-HKU and WHO-Hamburg RT-PCR assays was determined to be 0.1 50% tissue culture infective dose. Using seroconversion to CoV as the "gold standard" for SARS CoV diagnosis, WHO-HKU and WHO-Hamburg RT-PCR assays exhibited diagnostic sensitivities of 61 and 68% (nasopharyngeal aspirate specimens), 65 and 72% (throat swab specimens), 50 and 54% (urine specimens), and 58 and 63% (stool specimens), respectively, with an overall specificity of 100%. For patients confirmed to have SARS CoV and from whom two or more respiratory specimens were collected, testing the second specimen increased the sensitivity from 64 and 71% to 75 and 79% for the WHO-HKU and WHO-Hamburg RT-PCR assays, respectively. Testing more than one respiratory specimen will maximize the sensitivity of PCR assays for SARS CoV.


Optically Triggered Stepwise Double-Proton Transfer in an Intramolecular Proton Relay: A Case Study of 1,8-Dihydroxy-2-naphthaldehyde.
1,8-Dihydroxy-2-naphthaldehyde (DHNA), having doubly intramolecular hydrogen bonds, was strategically designed and synthesized in an aim to probe a long-standing fundamental issue regarding synchronous versus asynchronous double-proton transfer in the excited state. In cyclohexane, DHNA shows the lowest lying S0 →S1 (π-π*) absorption at ∼400 nm. Upon excitation, two large Stokes shifted emission bands maximized at 520 and 650 nm are resolved, which are ascribed to the tautomer emission resulting from the first and second proton-transfer products, denoted by TA* and TB*, respectively. The first proton transfer (DHNA* → TA*) is ultrafast (< system response of 150 fs), whereas the second proton transfer is reversible, for which the rates of forward (TA* → TB*) and backward (TA* ← TB*) proton transfer were determined to be (1.7 ps)(-1) and (3.6 ps)(-1), respectively. The fast equilibrium leads to identical population lifetimes of ∼54 ps for both TA* and TB* tautomers. Similar excited-state double-proton transfer takes place for DHNA in a single crystal, resulting in TA* (560 nm) and TB* (650 nm) dual-tautomer emission. A comprehensive 2D plot of reaction potential energy surface further proves that the sequential two-step proton motion is along the minimum energetic pathway firmly supporting the experimental results. Using DHNA as a paradigm, we thus demonstrate unambiguously a stepwise, proton-relay type of intramolecular double-proton transfer reaction in the excited state, which should gain fundamental understanding of the multiple proton transfer reactions.


[Diagnostic difficulties in herpes simplex encephalitis patients: report of three cases].
Herpes simplex encephalitis (HSE) is a rare infection of the central nervous system. The prognosis is poor, even in those receiving anti-herpetic therapy (about 20% of patients die, and in 40% of cases neurological sequelae are severe). The diagnosis is based on clinical signs of encephalitis, typical pathological changes in MRI, and on the presence of anti-HSV antibody and/or HSV-DNA in the cerebrospinal fluid. Three cases of encephalitis are presented. The clinical course and pathological changes in the temporal lobes seen in MRI were characteristic of the HSV etiology. Early intrathecal synthesis of anty-HSV antibodies were not detected despite an increase in their serum level in the course of the disease. HSV-DNA detection with PCR was not performed. Nevertheless in all cases aciclovir therapy was introduced on the first day of hospitalization. Possibilities of diagnosing HSE on the basis of clinical signs and MRI scans during the first week of the disease are discussed, as well as the tissue whether the course of treatment would be changed accordingly to the PCR test results.


The use of brief psychotherapy with substance abusers in early recovery.
This preliminary clinical report describes the usefulness of brief individual psychotherapy as a technique to engage relapse-prone substance abusers in addiction treatment. The authors propose that Mann's model of brief psychotherapy in the context of Brown's developmental model of recovery can be used during the early stages of recovery to improve retention in treatment. The use of Mann's model involves the identification of a central issue that is related to the substance abuse. Six commonly seen central issues are discussed: negative affective states, traumatic events, character pathology, unresolved grief, developmental arrest, and persistent denial. It is around one of these issues that a brief psychotherapeutic intervention is structured.


Fundamental insights into the correlation between chromosome configuration and transcription.
Eukaryotic chromosomes exhibit a hierarchical organization that spans a spectrum of length scales, ranging from sub-regions known as loops, which typically comprise hundreds of base pairs, to much larger chromosome territories that can encompass a few mega base pairs. Chromosome conformation capture experiments that involve high-throughput sequencing methods combined with microscopy techniques have enabled a new understanding of inter- and intra-chromosomal interactions with unprecedented details. This information also provides mechanistic insights on the relationship between genome architecture and gene expression. In this article, we review the recent findings on three-dimensional interactions among chromosomes at the compartment, topologically associating domain, and loop levels and the impact of these interactions on the transcription process. We also discuss current understanding of various biophysical processes involved in multi-layer structural organization of chromosomes. Then, we discuss the relationships between gene expression and genome structure from perturbative genome-wide association studies. Furthermore, for a better understanding of how chromosome architecture and function are linked, we emphasize the role of epigenetic modifications in the regulation of gene expression. Such an understanding of the relationship between genome architecture and gene expression can provide a new perspective on the range of potential future discoveries and therapeutic research.


Biphasic external defibrillation for adults in ventricular fibrillation or pulseless ventricular tachycardia.
Cardiac arrest, as a result of ventricular fibrillation or pulseless ventricular tachycardia, is a common phenomenon, and the only treatment available is defibrillation. Currently, defibrillators deliver either a monophasic or biphasic shock, depending on the device used. In 2005, the American Heart Association published new cardiac arrest management guidelines, which included directions about energy selection for both types of defibrillators. These guidelines created a platform to address misconceptions that exist in the practice setting with regard to the use of biphasic defibrillators. The purpose of this literature review was to highlight the issues related to the practical use of biphasic energy, including optimal energy selection and safety.


[Effect of gamma radiation on the components of hepatitis B virus and the delta antigen].
The influence of ionizing irradiation on the serological activity of purified HBsAg, receptors to polymerized albumin, DNA-polymerase activity, delta agent, and delta antigen was studied. Different radiosensitivity of hepatitis B virus components and delta-system was demonstrated. The possibility of sterilization by irradiation of purified HBsAg preparations suitable for construction of a vaccine against hepatitis B was shown.


Prophylactic Treatment with Adlay Bran Extract Reduces the Risk of Severe Acute Radiation Dermatitis: A Prospective, Randomized, Double-Blind Study.
Acute radiation dermatitis is a frequent adverse effect in patients with breast cancer undergoing radiotherapy, but there are only a small number of studies providing evidence-based interventions for this clinical condition. Adlay is a cereal crop that has been previously shown to have anti-inflammatory and antioxidant properties. In this study, we seek to evaluate the effectiveness of oral prophylactic treatment with adlay bran extract in reducing the risk of severe acute radiation dermatitis. A total of 110 patients with breast cancer undergoing radiotherapy were analyzed. Using a prospective, randomized, double-blind design, 73 patients received oral treatment with adlay bran extract and 37 patients received olive oil (placebo). Treatment was started at the beginning of radiation therapy and continued until the termination of radiation treatment. Our results showed that the occurrence of severe acute radiation dermatitis (RTOG grade 2 or higher) was significantly lower in patients treated with oral adlay bran extract compared to placebo (45.2% versus 75.7%, adjusted odds ratio 0.24). No serious adverse effects from adlay bran treatment were noted. In conclusion, prophylactic oral treatment with adlay bran extract reduces the risk of severe acute radiation dermatitis and may have potential use in patients with breast cancer undergoing radiotherapy.


Pulmonary fibrosis in experimental acute respiratory disease.
Changes in collagen metabolism were examined in 3 models of acute respiratory disease in rats. Fibrotic changes in the lungs of rats were provoked by exposing them to paraquat (intraperitoneal), ozone (inhaled), or bleomycin (intratracheally injected). After an interval sufficient to allow histologically discernible fibrosis to occur (6 to 7 days), lungs were removed from the rats, and apparent collagen synthesis rates were determined with cultured lung minces incubated in medium containing 3H-proline. There was a significant increase (severalfold in the apparent collagen synthesis rates by lung minces from all the pneumotoxin-exposed rats in this study. Portions of the 3H-proline-labeled lung minces were then used for quantifying ratios of Type I to Type III collagen. Using CNBr mapping techniques and a combination of carboxymethylcellulose chromatography and polyacrylamide gel electrophoresis, we quantified Type I/Type III collagen for newly synthesized, 3H-labeled collagen as well as for total unlabeled collagen. In lung minces from normal rats, the ratio was 2:1 (65 to 70% Type I collagen) for both newly synthesized and total collagen. On the other hand, lung minces prepared from fibrotic rats accumulated a mixture of newly synthesized collagens that was substantially enriched for Type I collagen (80 to 85% Type I). There was no change in Type I/Type III collagen for total unlabeled collagen, nor was there any detectable increase of total collagen per lung after 1 wk. We conclude that an early event in experimental acute respiratory disease is a marked increase in the relative synthesis of Type I collagen; this shift occurs before there is observable increased accumulation of collagen in the lung.


Association between airflow limitation severity and arterial stiffness as determined by the brachial-ankle pulse wave velocity: a cross-sectional study.
Objective Chronic obstructive pulmonary disease (COPD) is often associated with concomitant systemic manifestations and comorbidities, such as cardiovascular disease. There are limited data regarding airflow limitation (AL) and atherosclerosis in Japanese patients, and the potential association between AL and arterial stiffness has not yet been investigated in Japanese patients. Therefore, the purpose of this study was to investigate the association between AL severity and arterial stiffness using the brachial-ankle pulse wave velocity (baPWV). Methods This cross-sectional study included 1,356 subjects aged 40-79 years without clinical cardiovascular diseases who underwent a comprehensive health screening that included spirometry, the baPWV measurement, and blood sampling during medical check-ups in 2009 at the Japanese Red Cross Kumamoto Health Care Center. AL was defined in accordance with the Global Initiative for COPD criteria (forced expiratory volume in one second / forced vital capacity of < 0.7). A cut-off baPWV value of >1,400 cm/s was used for risk prediction and screening. Results The average baPWV (SD) results were 1,578.0 (317.9), 1,647.3 (374.4), and 1,747.3 (320.1) cm/s in the patients with a normal pulmonary function, mild AL, and moderate-to-severe AL, respectively (p< 0.001). Using logistic regression models adjusted for the age, body mass index, smoking status, hypersensitive C-reactive protein levels, hypertension, hyperglycemia, and dyslipidemia, an increased baPWV (>1,400 cm/s) was significantly associated with moderate-to-severe AL compared with a normal pulmonary function (odds ratio=2.76; 95% confidence intervals, 1.37-5.55; p=0.004). Conclusion Our results indicated an association between AL and increased arterial stiffness. Arterial stiffness may therefore worsen with an increase in the severity of AL.


Assessment of the Second-Ionization Potential of Lawrencium: Investigating the End of the Actinide Series with a One-Atom-at-a-Time Gas-Phase Ion Chemistry Technique.
Experiments were performed at the Lawrence Berkeley National Laboratory 88-Inch Cyclotron facility to investigate the electron-transfer reduction reaction of dipositive Lr (Z = 103) with O2 gas. Ions of 255Lr were produced in the fusion-evaporation reaction 209Bi(48Ca,2n) 255Lr and were studied with a novel gas-phase ion chemistry technique. The produced 255Lr2+ ions were trapped and O2 gas was introduced, such that the charge-exchange reaction to reduce 255Lr2+ to 255Lr1+ was observed and the reaction rate constant was determined to be k = 1.5(7) × 10-10 cm3/mol/s. The observation that this reaction proceeds establishes the lower limit on the second ionization potential of Lr to be 13.3(3) eV. This gives further support that the actinide series terminates with Lr. Additionally, this result can be used to better interpret the situation concerning the placement of Lu and Lr on the periodic table within the current framework of the actinide hypothesis. The success of this experimental approach now identifies unique opportunities for future gas-phase reaction studies on actinide and super heavy elements.


[Diagnosis and treatment of aplastic anemia].
Severe aplastic anemia is a rare disorder in childhood. Among various therapeutical strategies bone marrow transplantation (BMT) and immunosuppressive treatment with antithymocyte globulin (ATG) have proven to be most successful. Priority should be given to BMT over ATG treatment for patients with HLA-identical donors. Patients with Fanconi's anemia require a reduced conditioning program with cyclophosphamide and irradiation for BMT. Own experiences indicate that cooperative studies are highly needed to improve medical care for patients with severe aplastic anemia.


[Injuries of the cervical spine--surgical therapy].
Surgical treatment of unstable cervical spine injuries has justifiably gained acceptance when based on definite indications and methods of internal fixation. Direct anterior screw fixation of the odontoid process is the appropriate procedure for stabilizing articulation fractures. Many instability problems of the upper cervical spine can be solved with posterior transarticular fixation C1/C2 according to Magerl. Of the posterior procedures for the lower cervical spine, hook-plate fixation according to Magerl is preferable to wirings. Plate fixation with hollow titanium screws (Morscher) has proved a successful anterior procedure. The two methods can be combined to treat severe osteoligamentous injuries.


Target-controlled Infusion Propofol Versus Sevoflurane Anaesthesia for Emergency Traumatic Brain Surgery: Comparison of the Outcomes.
The choice of anaesthetic techniques is important for the outcome of traumatic brain injury (TBI) emergency surgery. The objective of this study was to compare patient outcomes for target-controlled infusion (TCI) of propofol and sevoflurane anaesthesia. A total of 110 severe TBI patients, aged 18-60, who underwent emergency brain surgery were randomised into Group T (TCI) (n = 55) and Group S (sevoflurane) (n = 55). Anaesthesia was maintained in Group T with propofol target plasma concentration of 3-6 μg/mL and in Group S with minimum alveolar concentration (MAC) of sevoflurane 1.0-1.5. Both groups received TCI remifentanil 2-8 ng/mL for analgesia. After the surgery, patients were managed in the intensive care unit and were followed up until discharge for the outcome parameters. Demographic characteristics were comparable in both groups. Differences in Glasgow Outcome Scale (GOS) score at discharge were not significant between Group T and Group S (P = 0.25): the percentages of mortality (GOS 1) [27.3% versus 16.4%], vegetative and severe disability (GOS 2-3) [29.1% versus 41.8%] and good outcome (GOS 4-5) [43.6% versus 41.8%] were comparable in both groups. There were no significant differences in other outcome parameters. TCI propofol and sevoflurane anaesthesia were comparable in the outcomes of TBI patients after emergency surgery.


[A new oxytocin infusion rate recorder (author's transl)].
A new oxytocin-infusion rate recorder system is introduced. It provides continuous plotting dosages on the labor channel of the cardiotocogram. So far 82 conductions of labour were monitored with the new system and evaluated for oxytocin overstimulation. The rate of overstimulations could be reduced resulting in better fetal outcome.


Gas chromatography-mass spectrometry profiles of urinary organic acids in healthy captive cheetahs (Acinonyx jubatus).
In captivity, cheetahs (Acinonyx jubatus) frequently suffer from several unusual chronic diseases that rarely occur in their free-ranging counterparts. In order to develop a better understanding of their metabolism and health we documented the urine organic acids of 41 apparently healthy captive cheetahs, in an untargeted metabolomic study, using gas chromatography-mass spectrometry. A total of 339 organic acids were detected and annotated. Phenolic compounds, thought to be produced by the anaerobic fermentation of aromatic amino acids in the distal colon, as well as their corresponding glycine conjugates, were present in high concentrations. The most abundant organic acids in the cheetahs' urine were an as yet unidentified compound and a novel cadaverine metabolite, tentatively identified as N1,N5-dimethylpentane-1,5-diamine. Pantothenic acid and citramalic acid concentrations correlated negatively with age, while glutaric acid concentrations correlated positively with age, suggesting possible dysregulation of coenzyme A metabolism in older cheetahs. This study provides a baseline of urine organic acid reference values in captive cheetahs and suggests important avenues for future research in this species.


Novel mutations in genes causing hereditary spastic paraplegia and Charcot-Marie-Tooth neuropathy identified by an optimized protocol for homozygosity mapping based on whole-exome sequencing.
Homozygosity mapping is an effective approach for detecting molecular defects in consanguineous families by delineating stretches of genomic DNA that are identical by descent. Constant developments in next-generation sequencing created possibilities to combine whole-exome sequencing (WES) and homozygosity mapping in a single step. Basic optimization of homozygosity mapping parameters was performed in a group of families with autosomal-recessive (AR) mutations for which both single-nucleotide polymorphism (SNP) array and WES data were available. We varied the criteria for SNP extraction and PLINK thresholds to estimate their effect on the accuracy of homozygosity mapping based on WES. Our protocol showed high specificity and sensitivity for homozygosity detection and facilitated the identification of novel mutations in GAN, GBA2, and ZFYVE26 in four families affected by hereditary spastic paraplegia or Charcot-Marie-Tooth disease. Filtering and mapping with optimized parameters was integrated into the HOMWES (homozygosity mapping based on WES analysis) tool in the GenomeComb package for genomic data analysis. We present recommendations for detection of homozygous regions based on WES data and a bioinformatics tool for their identification, which can be widely applied for studying AR disorders.Genet Med 18 6, 600-607.


Implications of multiple mechanisms of carcinogenesis for short-term testing.
The attempt has been made recently to categorize carcinogens into two mechanistic types based on their mechanism of action: genotoxic (capable of reacting with and damaging DNA) and epigenetic (unable to damage DNA to any detectable extent). By requiring that a given chemical fit into one or the other of these narrowly defined categories for regulatory purposes, we are probably oversimplifying potential biological effects. In fact, based on our limited understanding of carcinogenic mechanisms, this artificial distinction should probably be abandoned in favor of a more precise statement of each chemical's mechanism or relative potency of initiating and promoting effects. Since the standard short-term tests by which carcinogenicity of chemicals is screened were designed to detect certain chemical classes with active electrophilic intermediates, weak or specialized carcinogens may be missed and may be assumed erroneously to be nongenotoxic. The mechanisms of carcinogenicity for such carcinogens may include particulate deposition, active radical formation, liver toxicity, and hormonal interactions. Not all of these secondary mechanisms depend upon a detectable level of binding to DNA, damage to DNA, or modification of the DNA sequence, even though they may demonstrate other characteristics of a complete carcinogen (that is, irreversibility and lack of a threshold). Certain agents have been labeled as epigenetic. However, a consideration of the literature on sample agents (diethylstilbestrol, asbestos, and urethane) reveals that these are not epigenetic carcinogens despite their being labeled as such. Agents with irreversibility and no threshold have initiating potential and, as such, are genotoxic, whereas carcinogens that are classified as nongenotoxic are largely agents that promote the growth of liver tumors. Even promotion can be a mechanistically specialized phenomenon. For example, some promoters are cytotoxic to the liver, but not all liver toxins are liver tumor promoters or liver carcinogens. Further, the carcinogens commonly labeled as epigenetic might cause a unique specialized genotoxicity not detected by common screening tests routinely used for detecting genotoxicity. If we assume that this unrecognized but necessary initiating potential is mediated by some specialized genotoxicity, extra care must be taken to establish a genuine lack of genotoxicity before an agent can be classified (and regulated) as a promoter (lacking the ability to initiate tumor growth but still enhancing tumor development).(ABSTRACT TRUNCATED AT 400 WORDS)


Motor vehicle collision-related injuries and deaths among Indigenous Peoples in Canada: Meta-analysis of geo-structural factors.
Indigenous Peoples are much more likely than non-Indigenous Peoples to be seriously injured or die in motor vehicle collisions (MVCs). This study updates and extends a previous systematic review, suggesting that future research ought to incorporate social-environmental factors. We conducted a systematic review and meta-analysis of the published and grey literature on MVCs involving Indigenous Peoples in Canada between 2010 and 2020. We focussed on personal (e.g. driving an old vehicle) and community social-environmental-economic factors (e.g. prevalent low socioeconomic status). Eleven comparative cohorts that resulted in 23 at minimum, age-standardised, mortality or morbidity rate outcomes were included in our meta-analysis. Indigenous Peoples were twice as likely as non-Indigenous Peoples to be seriously injured (rate ratio [RRpooled] = 2.18) and more than 3 times as likely to die (RRpooled = 3.40) in MVCs. Such great risks to Indigenous Peoples do not seem to have diminished over the past generation. Furthermore, such risks were greater on-reserves and in smaller, rural and remote, places. Such places may lack community resources, including fewer transportation and healthcare infrastructural investments, resulting in poorer road conditions in Indigenous communities and longer delays to trauma care. This seems to add further evidence of geo-structural violence (geographical and institutional violence) perpetrated against Indigenous Peoples in yet more structures (i.e. institutions) of Canadian society. Canada's system of highways and roadways and its remote health-care system represent legitimate policy targets in aiming to solve this public health problem.


Complications of computer-assisted navigation in total knee replacement: retrospective cohort of eight hundred and seventy eight consecutive knees.
The main objective was to analyze the computer-assisted navigation (CAN)-specific complications that forced to switch to conventional procedure in primary total knee replacement (TKR). The secondary objective was to determine the influence of those complications on TKR short-term survival. Retrospective study of 878 primary TKR in 753 patients. Two consecutive versions of the OrthoPilot navigation system (Braun Aesculap, Germany) were used during the study time. Specific complications of CAN were defined as those due to the instrumentation (hardware or software failures), which were classified in one of two categories according to whether they occurred during the registration or tracking process. There were 20 (2.3%) complications related to the navigation system use that forced to switch to conventional surgery: in 11 (1.2%) knees due to loosening of the tracking pins, and in the other nine (1.0%) there were information system failures. There was a trend for a higher conversion rate to conventional surgery with the use of the first version of the software. There were no fractures, infections, or nerve injuries at the pin site. We found no differences in the distribution of baseline variables among those with or without conversion to conventional surgery. There was no significant difference (p = 0.488) in the two year survival between patients with or without conversion. CAN for primary TKR is a safe method with few specific complications that forced to switch from the navigated to the conventional procedure. Conversion to conventional surgery did not affect the short-term survival of TKR.


Two new arylsulfatase A (ARSA) mutations in a juvenile metachromatic leukodystrophy (MLD) patient.
Fragments of the arylsulfatase A (ARSA) gene from a patient with juvenile-onset metachromatic leukodystrophy (MLD) were amplified by PCR and ligated into MP13 cloning vectors. Clones hybridizing with cDNA for human ARSA were selected, examined for appropriate size inserts, and used to prepare single-stranded phage DNA. Examination of the entire coding and most of the intronic sequence revealed two putative disease-related mutations. One, a point mutation in exon 3, resulted in the substitution of isoleucine by serine. Introduction of this alteration into the normal ARSA cDNA sequence resulted in a substantial decrease in ARSA activity on transient expression in cultured baby hamster kidney cells. About 5% of the control expression was observed, suggesting a small residual activity in the mutated ARSA. The second mutation, a G-to-A transition, occurred in the other allele and resulted in an altered splice-recognition sequence between exon 7 and the following intron. The mutation also resulted in the loss of a restriction site. Apparently normal levels of mRNA were generated from this allele, but no ARSA activity or immuno-cross-reactive material could be detected. A collection of DNA samples from known or suspected MLD patients, members of their families, and normal controls was screened for these mutations. Four additional individuals carrying each of the mutations were found among the nearly 100 MLD patients in the sample. Gene segregation in the original patient's family was consistent with available clinical and biochemical data. No individuals homozygous for either of these two mutations were identified. However, combinations with other MLD mutations suggest that the point mutation in exon 3 does result in some residual enzyme activity and is associated with late-onset forms of the disease. The splice-site mutation following exon 7 produces late-infantile MLD when combined with other enzyme-null mutations, implying that it is completely silent enzymatically.


Single nucleotide polymorphisms of the haptoglobin gene in non-small cell lung cancer treated with personalized peptide vaccination.
The present study analyzed polymorphisms of the 5' flanking region (from nt -840 to +151) of the haptoglobin gene in 120 patients with advanced non-small cell lung cancer (NSCLC) receiving personalized peptide vaccinations. In the region, six single nucleotide polymorphisms (SNPs) were confirmed, of which two, rs5472 and rs9927981, were completely linked to each other. The minor allele frequencies of rs5472/rs9927981 and rs4788458 were higher than those of the other three SNPs. The genotype frequencies of rs5472 or rs9927981 were A/A or C/C (42.5%, n=51), A/G or C/T (40.8%, n=49), and G/G or T/T (16.7%, n=20), respectively; and those of rs4788458 were T/T (34.2%, n=41), T/C (40.0%, n=48), and C/C (25.8%, n=31). The association between polymorphism rs5472/rs9927981 and prognosis, or between rs4788458 and prognosis, was analyzed further. However, no correlation was found between these SNPs and overall survival, regardless of subgroup analysis of gender, histology or concurrent therapy. These results suggest that the polymorphisms rs5472/rs9927981 and rs4788458 are not useful prognostic tools for patients with NSCLC treated with personalized peptide vaccination.


Effect of bacteriophage infection in combination with tobramycin on the emergence of resistance in Escherichia coli and Pseudomonas aeruginosa biofilms.
Bacteriophage infection and antibiotics used individually to reduce biofilm mass often result in the emergence of significant levels of phage and antibiotic resistant cells. In contrast, combination therapy in Escherichia coli biofilms employing T4 phage and tobramycin resulted in greater than 99% and 39% reduction in antibiotic and phage resistant cells, respectively. In P. aeruginosa biofilms, combination therapy resulted in a 60% and 99% reduction in antibiotic and PB-1 phage resistant cells, respectively. Although the combined treatment resulted in greater reduction of E. coli CFUs compared to the use of antibiotic alone, infection of P. aeruginosa biofilms with PB-1 in the presence of tobramycin was only as effective in the reduction of CFUs as the use of antibiotic alone. The study demonstrated phage infection in combination with tobramycin can significantly reduce the emergence of antibiotic and phage resistant cells in both E. coli and P. aeruginosa biofilms, however, a reduction in biomass was dependent on the phage-host system.


Clinical outcomes at 12 months and risk of inflammatory bowel disease in patients with an intermediate raised fecal calprotectin: a 'real-world' view.
A recent systematic review confirmed the usefulness of fecal calprotectin (FC) in distinguishing organic (inflammatory bowel disease (IBD)) from non-organic gastrointestinal disease (irritable bowel syndrome (IBS)). FC levels <50 μg/g have a negative predictive value >92% to exclude organic gastrointestinal (GI) disease. Levels >250 μg/g correlate with endoscopic IBD disease activity; sensitivity 90%. We aimed to determine clinical outcomes in intermediate raised FC results (50-250 μg/g). Primary care general practices in Coventry and Warwickshire, and 3 secondary care hospitals. 443 FC results in adults (>16 years old) were reviewed from July 2012 to October 2013. Clinical data was collected from hospital databases and general practitioners. Long-term clinical data was available in 41 patients (out of 48). The number of new diagnoses of IBD, IBS and other diagnoses for the intermediate group. The number referred and discharged from secondary care. A new IBD diagnosis was made in 19% (n=8) of intermediate results (1% of normal and 38% of raised results). 5% (n=2) of intermediate results had known IBD in remission. A new IBS diagnosis was made in 27% (n=11) of intermediate results, while 34% (n=14) remained undiagnosed, although 8 of these were not referred to secondary care. FC testing remains useful in aiding diagnosis of organic GI conditions. However, unlike negative and strongly positive FC results, intermediate FC results lead to a mixture of diagnoses. The OR of a new diagnosis of IBD for an intermediate result compared to normal FC result was 26.6, while an intermediate FC result gave an OR of 0.54 for a new IBS diagnosis compared to normal FC. For intermediate FC results, 1 in 3 patients remained in secondary care after 12 months with an OR of 3.6 compared to a normal FC result.


Proteome of endothelial cell-derived procoagulant microparticles.
Microparticles (MP) are small membrane vesicles that are released from cells upon activation or during apoptosis. Cellular MP in body fluids constitute a heterogeneous population, differing in cellular origin, numbers, size, antigenic composition and functional properties. MP support coagulation by exposure of tissue factor (TF), the initiator of coagulation in vivo. Moreover, MP may transfer bioactive molecules to other cells, thereby stimulating them to produce cytokines, cell-adhesion molecules, growth factors and TF, and modulate endothelial functions. However, a comprehensive characterization of the antigenic composition of MP has been poorly defined. This study describes the protein composition of endothelial cell (EC)-derived MP (EMP) using a proteomic approach. MS analysis indicated the presence of newly described protein such as metabolic enzymes, proteins involved in adhesion and fusion processes, members of protein folding event, cytoskeleton associated proteins and nucleosome. In conclusion, circulating EMP behave as an actual storage pool, able to disseminate blood-borne TF activity and other bioactive effectors, as confirmed by our experiments showing an increased procoagulant activity of EC exposed to EMP.


Effect of Whole Body Hypothermia on Surfactant Function When Amniotic Fluid Is Meconium Stained.
We provide the first complete biophysical study of surfactant by captive bubble surfactometry from a neonate with meconium-stained amniotic fluid under controlled whole body hypothermia. Surfactant function improves after 48 hours of hypothermia, as surfactant cholesterol decreases. These findings partially explain positive effect of hypothermia on respiratory outcomes during meconium aspiration syndrome. A larger study including several neonates with or without lung disease is being conducted to better define the effect of hypothermia on surfactant function.


Clinical effectiveness and clinical toxicity associated with platinum-based doublets in the first-line setting for advanced non-squamous non-small cell lung cancer in Chinese patients: a retrospective cohort study.
Real-world evidence lacks for clinical effectiveness and clinical toxicity associated with platinum-based doublets in the first-line setting for advanced non-squamous non-small cell lung cancer (advNS-NSCLC) in Chinese patients. Patients receiving first-line chemotherapy for advNS-NSCLC in four Chinese tertiary care hospitals from 2007 to 2012 were retrospectively identified for chart review. Propensity score methods created best matched pairs for platinum/pemetrexed versus other platinum-based doublets for head-to-head comparisons of early treatment discontinuation (completed treatment cycles <4), treatment failure (progressive disease or early treatment discontinuation), and adverse events (AE). Conventional multiple logistic regression analyses were also performed to confirm the impact of the studied platinum-based doublets on early treatment discontinuation, treatment failure, and hematological AE using vinorelbine/platinum as reference. 1,846 patients were included to create propensity score matched treatment groups for platinum/pemetrexed versus docetaxel (95 pairs), paclitaxel (118 pairs), gemcitabine (199 pairs), and vinorelbine (72 pairs)-contained doublet, respectively. Platinum/pemetrexed was associated with significantly lower risks of early treatment discontinuation (odds ratio (OR) ranged from 0.239, p = 0.001 relative to platinum/docetaxel to 0.389, p = 0.003 relative to platinum/paclitaxel) and treatment failure (OR ranged from 0.257, p < 0.001 relative to platinum/paclitaxel to 0.381, p < 0.001 relative to platinum/gemcitabine) than the other four studied doublets. Platinum/pemetrexed was also associated with several significantly lower hematological AE rates, such as versus platinum/paclitaxel (any hematological AE: OR 0.508, p = 0.032), platinum/gemcitabine (i.e., any hematological AE: OR 0.383, p < 0.001; anemia: OR 0.357, p < 0.001; thrombocytopenia: OR 0.345, p < 0.001) or platinum/vinorelbine (i.e., neutropenia: OR 0.360, p = 0.046; anemia: OR 0.181, p = 0.014) in matched patients. Further conventional logistic regression analyses indicated that pemetrexed/platinum was ranked lowest for the risks of early treatment discontinuation (OR 0.326, p < 0.001), treatment failure (OR 0.460, p < 0.001), and any hematological AE (OR 0.329, p < 0.001). Pemetrexed plus platinum had significantly superior clinical effectiveness as compared to the other platinum-based doublets with third-generation cytotoxic agents and was also associated with several lower hematological toxicity rates than gemcitabine or vinorelbine-based doublet in the first-line setting for advNS-NSCLC in Chinese patients.


A reanalysis of cluster randomized trials showed interrupted time-series studies were valuable in health system evaluation.
There is often substantial uncertainty about the impacts of health system and policy interventions. Despite that, randomized controlled trials (RCTs) are uncommon in this field, partly because experiments can be difficult to carry out. An alternative method for impact evaluation is the interrupted time-series (ITS) design. Little is known, however, about how results from the two methods compare. Our aim was to explore whether ITS studies yield results that differ from those of randomized trials. We conducted single-arm ITS analyses (segmented regression) based on data from the intervention arm of cluster randomized trials (C-RCTs), that is, discarding control arm data. Secondarily, we included the control group data in the analyses, by subtracting control group data points from intervention group data points, thereby constructing a time series representing the difference between the intervention and control groups. We compared the results from the single-arm and controlled ITS analyses with results based on conventional aggregated analyses of trial data. The findings were largely concordant, yielding effect estimates with overlapping 95% confidence intervals (CI) across different analytical methods. However, our analyses revealed the importance of a concurrent control group and of taking baseline and follow-up trends into account in the analysis of C-RCTs. The ITS design is valuable for evaluation of health systems interventions, both when RCTs are not feasible and in the analysis and interpretation of data from C-RCTs.


Effects of freezing and drying grass products prior to fatty acid extraction on grass fatty acid and lipid class composition--a technical note.
Grass and grass silage represent a rich and natural source of omega-3 polyunsaturated fatty acids, in particular linolenic acid, for ruminants. Recent research, focusing on improving the content of these beneficial fatty acids in grass, requires storage of the forage samples prior to analysis. In this study, we evaluated whether conservation of fresh grass and grass silage by freezing (1 and 4 weeks,--18 degrees C) and/or drying (24h, 50 degrees C) affected its fatty acid content and induced shifts between lipid classes. FA were extracted using chloroform/methanol (2/1, v/v) and triacylglycerols (TAG), free fatty acids (FFA) and polar lipids (PL) were separated by thin layer chromatography. Fatty methyl esters (FAME) were identified by gas chromatography. Loss of thawing liquor might provoke a dramatic decrease in extractable lipid after frozen storage of both grass and grass silage. Morever, after frozen storage, fatty acids in grass but not in grass silage seem subjected to a higher rate o f lipolysis and oxidation, as suggested by increased quantities of FFA (3.1, 7.6, 8.4 % of total FAME) and reduced proportions of poly-unsaturated fatty acids (79.5, 73.6 and 74.1 % of total FAME) when analysing fresh grass samples directly or after 1 and 4 weeks of frozen storage, respectively. Drying of fresh grass did not provoke changes in FA composition, but distribution of FA over lipid classes was significantly altered, with an increase in TAG (5.1 to 17.9 % of total FAME) and FFA (2.4 to 14.9 % of total FAME) and lower proportions of PL (90.7 to 55.7 % of total FAME).


SETDB1 accelerates tumourigenesis by regulating the WNT signalling pathway.
We investigated the oncogenic role of SETDB1, focusing on non-small cell lung cancer (NSCLC), which has high expression of this protein. A total of 387 lung cancer cases were examined by immunohistochemistry; 72% of NSCLC samples were positive for SETDB1 staining, compared to 46% samples of normal bronchial epithelium (106 cases) (p <0.0001). The percentage of positive cells and the intensity of staining increased significantly with increased grade of disease. Forced expression of SETDB1 in NSCLC cell lines enhanced their clonogenic growth in vitro and markedly increased tumour size in a murine xenograft model, while silencing (shRNA) SETDB1 in NSCLC cells slowed their proliferation. SETDB1 positively stimulated activity of the WNT-β-catenin pathway and diminished P53 expression, resulting in enhanced NSCLC growth in vitro and in vivo. Our finding suggests that therapeutic targeting of SETDB1 may benefit patients whose tumours express high levels of SETDB1.


Metformin improves cardiac function in rats via activation of AMP-activated protein kinase.
1. Metformin is one of the most commonly used drugs for the treatment of Type 2 diabetes. Accumulating evidence suggests that metformin also has cardioprotective effects. In the present study, we investigated the cardioprotective effects of metformin and the mechanisms involved. 2. A rat model of chronic heart failure was established by permanent left coronary artery occlusion. Heart failure rats were randomly divided into four groups: (i) a saline-treated group given 4 mL/kg day via intragastric gavage; (ii) a metformin-treated group, given 100 mg/kg metformin once daily via intragastric gavage; (iii) a group treated with 5 mg/kg 5'-aminoimidazole-4-carboxyamide-ribonucleoside (AICAR), an AMP-activated protein kinase (AMPK) agonist, every second day; and (iv) a group treated with 100 mg/kg per day metformin + 20 mg/kg, i.p., compound C (an AMPK antagonist). After 4 weeks treatment, echocardiography was used to assess left ventricular (LV) dimensions and function. Expression of AMPK, endothelial nitric oxide synthase (eNOS) and transforming growth factor (TGF)-β1 was determined by reverse transcription-polymerase chain reaction and western blot analysis. 3. Metformin administration significantly improved cardiac function and LV remodelling, as evidenced by increases in LV systolic pressure and LV ejection fraction and decreases in LV end-diastolic diameter and LV end-systolic diameter. These beneficial effects of metformin were associated with increased AMPK and eNOS phosphorylation, as well as reductions in insulin, TGF-β1, basic fibroblast growth factor and tumour necrosis factor-α levels in the circulation and/or myocardium. 4. The results indicate that chronic low-dose metformin confers significant cardioprotective effects against chronic heart failure by activating the AMPK-eNOS pathway.


Comparison of Clinical Effects of Temozolomide Single Agent and Combined Doxorubicin in the Treatment of Glioma.
In this article, we have compared and analyzed the clinical effects of temozolomide single agent and combined doxorubicin in the treatment of glioma. To evaluate this, a total of 70 patients diagnosed with glioma in our hospital from July 2019 to July 2020 were randomly divided into two groups, the observation group and the control group, with 35 patients in each group. The control group received temozolomide capsules orally. The observation group was treated with temozolomide single agent and doxorubicin. After treatment, the clinical efficacy, adverse reactions, and KPS score of the two groups were observed. After treatment, the total response rate of the control group was 31.43%, and the total response rate of the observation group was 62.86%. The difference between the two groups was statistically significant (P < 0.05). Before treatment, there was no significant difference in KPS scores between the two groups (P > 0.05). After treatment, the KPS scores of both groups were improved, and the KPS scores of the observation group and the control group were significantly better, with statistical significance (P < 0.05). In the observation group, 17 cases had adverse reactions, including 10 cases of nausea and vomiting, 2 cases of leucopenia, and 5 cases of thrombocytopenia, with a total incidence of 48.57%. In the control group, there were 31 cases of adverse reactions, including 22 cases of nausea and vomiting, 6 cases of leucopenia, and 4 cases of thrombocytopenia, with a total incidence of 91.43%. The difference between the two groups was statistically significant (P < 0.05). The efficacy of temozolomide single agent and combined doxorubicin in the treatment of glioma was significant. Moreover, it can significantly improve clinical efficacy, reduce the incidence of adverse reactions, and improve the health status of patients, which is worthy of further clinical application.


Cardiopulmonary Resuscitation Training in Schools Following 8 Years of Mandating Legislation in Denmark: A Nationwide Survey.
School cardiopulmonary resuscitation (CPR) training has become mandatory in many countries, but whether legislation has translated into implementation of CPR training is largely unknown. We assessed CPR training of students following 8 years of legislative mandates in Denmark. A nationwide cross-sectional survey of Danish school leadership (n=1240) and ninth-grade homeroom teachers (n=1381) was carried out for school year 2013-2014. Qualitative interviews and the Theory of Planned Behavior were used to construct the survey. Logistic regression models were employed to identify factors associated with completed CPR training. Information from 63.1% of eligible schools was collected: 49.3% (n=611) of leadership and 48.2% (n=665) of teachers responded. According to teachers, 28.4% (95% CI 25.0% to 32.0%) and 10.3% (95% CI 8.1% to 12.8%) of eligible classes had completed CPR and automated external defibrillator training, respectively. Among leadership, 60.2% (95% CI 56.2% to 64.1%) reported CPR training had occurred during the 3 years prior to the survey. Factors associated with completed CPR training included believing other schools were conducting training (odds ratio [OR] 9.68 [95% CI 4.65-20.1]), awareness of mandating legislation (OR 4.19 [95% CI 2.65-6.62]), presence of a school CPR training coordinator (OR 3.01 [95% CI 1.84-4.92]), teacher feeling competent to conduct training (OR 2.78 [95% CI 1.74-4.45]), and having easy access to training material (OR 2.08 [95% CI 1.57-2.76]). Despite mandating legislation, school CPR training has not been successfully implemented. Completed CPR training was associated with believing other schools were conducting training, awareness of mandating legislation, presence of a school CPR training coordinator, teachers teacher feeling competent to conduct training, and having easy access to training material. Facilitating these factors may increase rates of school CPR training.


Surface roughness-induced absorption acts as an ovarian cancer cells growth sensor-monitor.
Uncontrolled growth of ovarian cancer cells is the fifth leading cause of female cancer deaths since most ovarian cancer patients are diagnosed at an advanced stage of metastatic disease. Here, we report on the sensor for monitoring the cancer treatment efficiency in real-time. We measure the optical interaction between the evanescent fields of microfiber and ovarian cancer inter-cellular medium at different treatment stages. Spectral absorption signatures are correlated with optical micrographs and western blot tests. We found that the treatment of tumor cells with induces both cells growth arrest and alter the spectral lines in a dose-dependent manner. These observations are mediated by surface roughness out of silica glass material, form an essential step toward the development of early detection of response to cancer therapy.


Histopathology and external examination of heavily parasitized Lost River Sucker Deltistes luxatus (Cope 1879) and Shortnose Sucker Chasmistes brevirostris (Cope 1879) from Upper Klamath Lake, Oregon.
Shortnose Sucker (Chasimistes brevirostris) and Lost River Sucker (Deltistes luxatus) are endemic to the Upper Klamath Basin of Southern Oregon and Northern California, and their populations are in decline. We used histopathology and external examination of 140 and external examination only of 310 underyearling suckers collected in 2013, 2015 and 2016 to document pathological changes, particularly those relating to parasites. The most severe infection was caused by a Contracaecum sp., infecting the atrium of 8%-33% of Shortnose Suckers. The most prevalent infections were caused by Bolbophorus sp. metacercariae in the muscle of Shortnose Suckers (21%-63%) and Lernaea cyprinacea in the skin and muscle of Lost River Suckers (30%-81%). Histology detected Bolbophorus in only 5% of cases where it was not seen externally. Three myxozoans were observed; a Parvicapsula sp. in the renal tubules (10%), a Myxobolus sp. in the intestinal mucosa (2%) and an unusual multicellular, presporogonic myxozoan in the intestinal lumen of one sucker. Severe gill epithelial hyperplasia was observed in several fish collected in 2016. Trichodinids and Ichthyobodo sp. were observed on some of the gills, but absent in many of the fish with severe lesions. A histiocytic sarcoma was observed in sucker.


Blood glucose determinations in newborns: four instruments compared.
Four portable analyzers, HemoCue B-Glucose (I), Accu-Check III (II), One-touch II (III), and Glucometer Elite (IV), with different measuring principles were tested for their suitability for measuring blood glucose in neonates. Precision of all instruments is satisfactory. In the analysis of capillary blood from newborns, two instruments show an excellent accuracy; however, the scatter of the results for instrument (II) is about 1.6 times greater than for instrument (I). The inaccuracy of instruments (III) and (IV) is not acceptable from a clinical point of view. All devices show an influence of hematocrit, the magnitude of which varies between 5% (I) and 12% (III) for every 10% change of hematocrit. Instruments II and IV show that temperature has a marked influence on the readings; the same is true for oxygen in instrument IV. In conclusion, only instrument (I) has met the requirements of accurate and precise blood glucose determinations in neonates.


Mifepristone: contraceptive and non-contraceptive uses.
Mifepristone is an orally active progesterone antagonist. It can be used for both contraceptive and non-contraceptive clinical indications. It is a very effective drug for emergency contraception with a low incidence of side effects. There is a potential for mifepristone to be used as a once-a-month pill. There is a need, however, for a simple, inexpensive and accurate method to identify the luteinizing hormone surge before this method can be used in clinical practice. The daily administration of mifepristone offers promise as an effective method of contraception but more studies need to be done. The combination of mifepristone with a prostaglandin analogue is a well-established method for termination of pregnancy of up to 9 weeks. Recent data suggest that this combination may also be used up to 9-13 weeks of pregnancy. Although mifepristone is effective in dilating the cervix before vacuum aspiration, misoprostol is probably the drug of choice in most situations. In the second trimester, mifepristone is effective in shortening the abortion process induced by prostaglandin analogues. The combination of mifepristone and prostaglandin also offers a medical method for management of miscarriages. Mifepristone has been used for a number of other indications, but further studies are needed before such treatment can be recommended.


Singularity solution of Lanir's osmoelasticity: verification of discontinuity simulations in soft tissues.
The literature characterizes cartilaginous tissues as osmoviscoelastic. Understanding the damage and failure of these tissues is essential for designing treatments. To determine tissue strength and local stresses, experimental studies-both clinical and animal-are generally supported by computational studies. Verification methods for computational studies of ionized porous media including cracks are hardly available. This study provides a method for verification and shows its performance. For this purpose, shear loading of a finite crack is addressed analytically and through a commercial finite element code. Impulsive shear loading by two-edge dislocation of a crack was considered in a 2D plane strain model for an ionized porous medium. To derive the analytical solution, the system of equation is decoupled by stress functions. The shear stress distribution at the plane of the crack is derived using Fourier and Laplace transformations. The analytical solution for the shear stress distribution is compared with computer simulations in ABAQUS version 6.4-5. Decoupling of the equations makes it possible to solve some boundary value problems in porous media taking chemical effects into account. The numerical calculations underestimate the shear stress at the crack-tips. Mesh refinement increases accuracy, but is still low in the neighborhood of the crack-tips.


Age and Social Support Seeking: Understanding the Role of Perceived Social Costs to Others.
We examined age differences in the use of different types of social support and the reasons for these differences. We found that older adults (age 60+) seek explicit social support less compared with young adults (age 18-25), but there is no difference in implicit social support seeking. Concerns about the potential social costs of seeking explicit support mediate the age differences in explicit social support seeking. Whereas young adults view this strategy as conferring more benefits than costs, older adults have a more balanced view of the costs and benefits of explicit social support seeking. Older and young adults do not differ in perceptions of the relative costs versus benefits of implicit social support seeking. Finally, we found older adults benefit more from implicit (vs. explicit) social support emotionally than young adults, which further explains why age groups differ in their use of explicit versus implicit social support.


Prospective association between adherence to the Mediterranean diet and risk of depressive symptoms in the French SU.VI.MAX cohort.
This study examines whether adherence to the Mediterranean Diet (MD) measured by several dietary indexes was associated with incident depressive symptoms in a large French cohort. The study sample consisted of 3523 participants from the Supplémentation en Vitamines et Minéraux Antioxydants (SU.VI.MAX) cohort who had at least three dietary records at baseline during the first 2 years of follow-up (1994-1996), free of depression at the beginning of the study (1996-1997) and available Center for Epidemiologic Studies Depression Scale (CES-D) data at the end of follow-up (2007-2009). The rMED was computed. Incident depressive symptoms were defined by a CES-D score ≥17 for men and ≥23 for women in 2007-2009. Odds ratios (OR) and 95% confidence intervals (95% CI) were estimated using multivariable logistic regression models. Several sensitivity analyses were performed. In the present study, 172 incident cases of depressive symptoms were identified during the follow-up (mean = 12.6 years). After adjustment for a wide range of potential confounders, adherence to the rMED score (continuous variable) was significantly associated with incident depressive symptoms in men (OR 0.91; 95% CI 0.83-0.99; p = 0.03), but not in women. Use of the Literature-Based Adherence Score to the Mediterranean Diet (LAMD) and the classic MD score (MDS) provide similar findings. In the current study, higher adherence to the Mediterranean Diet at midlife was associated with a lower risk of incident depressive symptoms, particularly in men, increasing scientific evidence for a beneficial role of Mediterranean Diet on health. Further investigations in particular among women are needed.


Combinatorial chemistry in food research.
This mini-review is concerned with emerging applications of combinatorial chemistry relevant to the needs of the food industry. More specifically, recent advances in the use of combinatorial methods for the identification and analysis of flavours, "food chemical" libraries as a potential source of enzyme inhibitors for the food industry and the utility of biocatalysis for the generation of molecular diversity are discussed.


The new evidence on cardiorenal benefits of sodium-glucose cotransporter 2 inhibitors in type 1 diabetes: one step closer to their use in this setting?
Chronic kidney disease (CKD) affects 30-40% of persons with type 1 diabetes mellitus (T1DM), markedly increasing the risk of kidney failure and cardiovascular events. The excessive mortality observed in T1DM compared to the general population can be attributed to the presence of CKD, with cardiovascular disease as the leading cause of premature death. A recently published, robust real-world study investigated the impact of sodium-glucose cotransporter 2 inhibitors (SGLT2i) and glucagon-like peptide-1 receptor agonists (GLP-1 RA) adjunctive therapy on blood glucose levels, adverse events, and cardio-renal outcomes among individuals with T1DM. GLP-1 RA provided greater reduction in glycated hemoglobin in comparison to SGLT2i therapy, whereas the latter reduced the risk of CKD, heart failure, and hospitalization for any cause. However, the SGLT2i treated cohort had a higher risk of diabetic ketoacidosis (DKA). The study provides promising evidence that the protective cardiorenal effects of SGLT2i, previously confirmed in people with and without type 2 diabetes mellitus, might also be present in T1DM. However, the benefits and risks, especially the risk of DKA, should be further examined in dedicated large-scale randomized controlled trials.


Stereochemistry in the synthesis and reaction of exo-glycals.
Two general methods are explored for the stereoselective synthesis of exo-glycals. One method utilizes a nucleophilic addition of fully protected sugar lactones of gluco-, galacto-, and manno-types, followed by the subsequent dehydration, to give the desired exo-glycals with (Z)-configuration. The other method proceeds with selenylation of C-glycosides in a stereoselective manner. The subsequent selenoxide elimination also provides (Z)-exo-glycals. The prepared exo-glycal conjugated esters of either gluco- or manno-type react with allyl alcohol to give exclusively alpha-anomers.


Dose dependence of CTL precursor frequency induced by a DNA vaccine and correlation with protective immunity against influenza virus challenge.
Intramuscular injection of BALB/c mice with a DNA plasmid encoding nucleoprotein (NP) from influenza virus A/PR/8/34 (H1N1) provides cross-strain protection against lethal challenge with influenza virus A/HK/68 (H3N2). CTL specific for the H-2Kd-restricted epitope NP147-155 are present in these mice and are thought to play a role in the protection. To assess the effectiveness of NP DNA immunization in comparison with influenza virus infection in the induction of CTL responses, we monitored the frequency of CTL precursors (CTLp) in mice following i.m. injection with NP DNA or intranasal infection with influenza virus and showed that the CTLp frequency in NP DNA-immunized mice can reach levels found in mice that had been infected with influenza virus. We also measured the CTLp frequency, anti-NP Ab titers, and T cell proliferative responses in mice that were injected with titrated dosages of NP DNA and documented a correlation of the CTLp frequency and the Ab titers, but not proliferative responses, with the injection dose. Furthermore, we observed a positive correlation between the frequency of NP147-155 epitope-specific CTLp and the extent of protective immunity against cross-strain influenza challenge induced by NP DNA injection. Collectively, these results and our early observations from adoptive transfer experiments of in vitro activated lymphocytes from NP DNA-immunized mice suggest a protective function of NP-specific CTLp in mice against cross-strain influenza virus challenge.


[Corneal transplantation at the beginning of the 21th century].
Corneal transplantation (keratoplasty) is necessary when various disorders result in corneal opacities with severe visual loss that cannot be treated conservatively, or the regular structure and curvature of the cornea is distorted, and its function is lost. Among human transplantation, keratoplasty is the most successful surgical procedure. In recent decades, penetrating keratoplasties have been increasingly replaced by lamellar techniques, where only the abnormal layer of the cornea is transplanted. The anterior form is deep anterior lamellar keratoplasty (DALK), recommended mainly for keratoconus. The main forms of posterior lamellar keratoplasty are Descemet's stripping automated endothelial keratoplasty (DSAEK) and Descemet's membrane endothelial keratoplasty (DMEK). The indications are bullous keratopathy after cataract surgery with endothelial destruction and Fuchs endothelial dystrophy. Lamellar keratoplasty has several advantages over penetrating surgery. Postoperative visual acuity is better, wound healing, patient rehabilitation are faster and the course of any immune rejection is milder and can be better managed. Orv Hetil. 2023; 164(28): 1087-1093.


Differing requirements for CCR4, E-selectin, and α4β1 for the migration of memory CD4 and activated T cells to dermal inflammation.
CCR4 on T cells is suggested to mediate skin homing in mice. Our objective was to determine the interaction of CCR4, E-selectin ligand (ESL), and α(4)β(1) on memory and activated T cells in recruitment to dermal inflammation. mAbs to rat CCR4 were developed. CCR4 was on 5-21% of memory CD4 cells, and 20% were also ESL(+). Anti-TCR-activated CD4 and CD8 cells were 40-55% CCR4(+), and ∼75% of both CCR4(+) and CCR4(-) cells were ESL(+). CCR4(+) memory CD4 cells migrated 4- to 7-fold more to dermal inflammation induced by IFN-γ, TNF, TLR agonists, and delayed-type hypersensitivity than CCR4(-) cells. CCR4(+) activated CD4 cells migrated only 5-50% more than CCR4(-) cells to these sites. E-selectin blockade inhibited ∼60% of CCR4(+) activated CD4 cell migration but was less effective on memory cells where α(4)β(1) was more important. Anti-α(4)β(1) also inhibited CCR4(-) activated CD4 cells more than CCR4(+) cells. Anti-E-selectin reduced activated CD8 more than CD4 cell migration. These findings modify our understanding of CCR4, ESL, α(4)β(1), and dermal tropism. There is no strict relationship between CCR4 and ESL for skin homing of CD4 cells, because the activation state and inflammatory stimulus are critical determinants. Dermal homing memory CD4 cells express CCR4 and depend more on α(4)β(1) than ESL. Activated CD4 cells do not require CCR4, but CCR4(+) cells are more dependent on ESL than on α(4)β(1), and CCR4(-) cells preferentially use α(4)β(1). The differentiation from activated to memory CD4 cells increases the dependence on CCR4 for skin homing and decreases the requirement for ESL.


[Clinical education in nursing].
There are two types of training programmes open to nurses to enable them to become a nurse clinician: certification-based programmes and masters degree. Based on the clinical nursing raisonning, these training courses enable professionals to develop their knowledge in nursing sciences and their expertise, notably in the framework of complex care pathways.


[Sero-epidemiological study of the hepatitis epidemic in Mitrovica in the aftermath of the war in Kosovo (1999)].
In July 1999, after the return of the Kosovar refugees to their country, an outbreak of acute hepatitis A (HAV) and E (HEV) occurred. Epidemic hepatitis and diarrhoea are closely monitored communicable diseases which had been singled out as priorities by the health authorities of the former Republic of Yougoslavia. Several field surveys were undertaken: description of reported cases, serological study of clinical patients, study of anti-HAV seroprevalence in a school and a case control study to assess risk factors. The analysis of the reports indicates an epidemic peak at the end of September, 2 or 3 weeks after the start of the new school year, with an increase of cases relative to the age of the children in school. In a serologic study of 104 samples, we found an anti-HAV IgM positivity in 88% of the cases. Children were more likely to be positive when compared to teenagers and adults; likewise, consumers of well-water as versus those drinking network water (p = 0.03). The study of seroprevalence showed that transmission had taken place within the school. The case control study, in spite of its imperfections, indicated that consumption of water melon, the only fruit cultivated locally, is a factor which facilitates the HAV transmission. Circulation of the hepatitis E virus was confirmed for 4 sick persons (including 2 co-infected HAV-HEV) and in 4 persons without clinical symptoms. These results show the permanent risk of faeco-oral infections in this area and the need to develop a relevant prevention policy.


Associations between biosecurity and outbreaks of canine distemper on Danish mink farms in 2012-2013.
During 8 months from July 2012 to February 2013, a major outbreak of canine distemper involving 64 mink farms occurred on the Danish peninsula of Jutland. The canine distemper outbreak was associated with exposure of farmed mink to infected wild carnivores and could represent a deficit in biosecurity on the mink farms. The aim of this study was to investigate the extent and association of specific biosecurity measures with the outbreak. The study was carried out in an epidemiological case-control design. The case group consisted of the 61 farms, which had a confirmed outbreak of canine distemper from July 2012 to February 2013. The control group included 54 farms without an outbreak of canine distemper in 2012 or 2013, selected as the closest geographical neighbour to a case farm. The results showed that significantly more control than case farms had vaccinated their mink against canine distemper virus. Mortality was only assessed on the case farms, and there was a non-significantly lower mortality on vaccinated farms than on the non-vaccinated farms. Furthermore, the proportion of farms with observations of wild red foxes (Vulpes vulpes) inside the farm enclosures were larger for case farms, indicating that the control farms had a better biosecurity or were not equally exposed to canine distemper virus. Generally, all farms had very few specific precautions at the gate entrance in respect to human visitors as well as animals. The use of biosecurity measures was very variable in both case and control farms. Not using plastic boot covers, presence of dogs and cats, presence of demarcated area for changing clothes when entering and leaving the farm area and presence of hand washing facilities significantly lowered the odds of the farm having a canine distemper virus outbreak. The results of the study indicate that consistent use of correct vaccination strategies, implementation of biosecurity measures and limiting human and animal access to the mink farm can be important factors in reducing the risk for canine distemper outbreaks.


Perinatal protein malnutrition alters expression of miRNA biogenesis genes Xpo5 and Ago2 in mice brain.
Due to its widespread incidence, maternal malnutrition remains one of the major non-genetic factors affecting the development of newborn's brain. While all nutrients have certain influence on brain maturation, proteins appear to be the most critical for the development of neurological functions. An increasing number of studies point out that the effects of early-life nutritional inadequacy has long lasting effects on the brain and lead to permanent deficits in learning and behavior. Epigenetic mechanisms provide a potential link between the nutrition status during critical periods and changes in gene expression that may lead to disease phenotypes. Among those epigenetic mechanisms microRNAs (miRNAs) emerge as promising molecules for the link between nutrition and gene expression due to their relevance in many central nervous system functions. The objective of the current study was to evaluate the impact of perinatal protein malnutrition on the development of male and female mice offspring and to analyze the expression of the genes involved in the miRNA biogenesis pathway in different mouse brain structures. We demonstrated that early nutritional stress such as exposition to a protein-deficient diet during gestation and lactation reduced the hippocampal weight, delayed offspring's development and deregulated the expression of Xpo5 and Ago2 genes in hippocampus and hypothalamus of weanling mice. Moreover, an overall increase in mature miRNAs was consistent with the induction of Xpo5 mRNA. Altered miRNA biogenesis could modify the availability and functionality of miRNA becoming a causal factor of the adverse effects of protein malnutrition.


A study of the relationship between cornea permeability and eye irritation using membrane-interaction QSAR analysis.
A methodology termed membrane-interaction QSAR (MI-QSAR) analysis has been used to develop QSAR models to predict drug permeability coefficients across cornea and its component layers (epithelium, stroma, and endothelium). From a training set of 25 structurally diverse drugs, significant QSAR models are constructed and compared for the permeability of the cornea, epithelium, and stroma plus endothelium. Cornea permeability is found to depend on the measured distribution coefficient of the drug, the cohesive energy of the drug, the total potential energy of the drug-membrane "complex," and three other energy refinement descriptor terms. The endothelium may be a more important barrier in cornea permeation than the stroma. Moreover, an investigation of the correlation between cornea permeation and eye irritation is presented as an example of a cross study on different ADMET properties using MI-QSAR analysis. Thirteen structurally diverse drugs, whose molar-adjusted eye irritation scores (MES) have been measured using the Draize rabbit-eye test, were chosen as an eye irritation comparison set. A poor correlation (R(2) = 0.0232) between the MES measures and the predicted cornea permeability coefficients for the drugs in the eye irritation set suggests there is no significant relationship between eye irritation potency and the cornea permeability.


[Relation between myocardial perfusion, myocardial necrosis and residual glucose metabolism as a vitality parameter in the post-ischemic myocardium following coronary reperfusion].
The assessment of myocardial viability following interventions aiming at restitution of antegrade blood flow by PTCA and thrombolysis has become one of the most important goals to judge interventional success. Conventional markers as ECG, the extent of the regional wall motion abnormality and enzymes have conceptual limitations. We had shown previously that increased residual glucose metabolism in reperfused myocardium can be assessed by F-18 2-deoxy-glucose (FDG) and positron emission tomography (PET) and will predict functional recovery in reperfused myocardium. The present study introduces a novel concept for the evaluation of myocardial viability by the application of multiple tracer techniques and investigates tracer behaviour in relation to blood flow, regional wall motion, coronary anatomy and histology. The experimental data compare segmental uptake of F-18 FDG with the current most clinically important perfusion marker Tl-201 and Tc-99m (Sn) pyrophosphate (Tc-99mPPi) as a marker of myocardial necrosis in a canine model following a 3 hour intracoronary balloon occlusion and subsequent reperfusion and therefore in a clinically attributable occlusion and reperfusion scenario. Myocardial blood flow was assessed by microspheres, regional fractional tracer retention fractions were evaluated from tissue tracer concentrations, the arterial input function and quantitative blood flow measurements. The extent of ischemic damage was evaluated by TTC post mortem staining, histology, and histochemistry. Additional studies to illucidate the mechanism of increased glucose utilization were done by analysis of other substrates as lactate. The result within the area at risk was assessed in relation to the metabolic behaviour. PET methods were validated in the same dog model as a quantitative biochemical in-vivo assay and imaging procedure and demonstrate the ability of this noninvasive techniques to measure blood flow and myocardial metabolism, especially exogenous glucose utilization. Time dependent and repetitive PET studies may indicate a delayed recovery of myocardial metabolism following an ischemic event with a pattern of a blood flow-metabolism mismatch, characteristic for persistent viability. Metabolic data by imaging correlate with reference techniques as arterio-venous sampling and post mortem histochemical staining method using TTC, PAS, and glycogen techniques. The clinical part applies the proposed tracer concept to patients with acute myocardial infarction and following reperfusion interventions as well using conventional markers and again positron markers. These studies indicate possibilities to assess myocardial viability by the same simultaneou


X-Cells Are Globally Distributed, Genetically Divergent Fish Parasites Related to Perkinsids and Dinoflagellates.
"X-cells" have long been associated with tumor-like formations (xenomas) in marine fish, including many of commercial interest. The name was first used to refer to the large polygonal cells that were found in epidermal xenomas from flatfish from the Pacific Northwest [1]. Similar looking cells from pseudobranchial xenomas had previously been reported from cod in the Atlantic [2] and Pacific Oceans [3]. X-cell pathologies have been reported from five teleost orders: Pleuronectiformes (flatfish), Perciformes (perch-like fish), Gadiformes (cods), Siluriformes (catfish), and Salmoniformes (salmonids). Various explanations have been elicited for their etiology, including being adenomas or adenocarcinomas [4, 5], virally transformed fish cells [6-8], or products of coastal pollution [9, 10]. It was hypothesized that X-cells were protozoan parasites [1, 11-13], and although recent molecular analyses have confirmed this, they have failed to place them in any phylum [14-18], demonstrating weak phylogenetic associations with the haplosporidians [16] or the alveolates [15]. Here, we sequenced rRNA genes from European and Japanese fish that are known to develop X-cell xenomas. We also generated a metagenomic sequence library from X-cell xenomas of blue whiting and Atlantic cod and assembled 63 X-cell protein-coding genes for a eukaryote-wide phylogenomic analysis. We show that X-cells group in two highly divergent clades, robustly sister to the bivalve parasite Perkinsus. We formally describe these as Gadixcellia and Xcellia and provide a phylogenetic context to catalyze future research. We also screened Atlantic cod populations for xenomas and residual pathologies and show that X-cell infections are more prevalent and widespread than previously known.


The promise of prevention: the effects of four preventable risk factors on national life expectancy and life expectancy disparities by race and county in the United States.
There has been substantial research on psychosocial and health care determinants of health disparities in the United States (US) but less on the role of modifiable risk factors. We estimated the effects of smoking, high blood pressure, elevated blood glucose, and adiposity on national life expectancy and on disparities in life expectancy and disease-specific mortality among eight subgroups of the US population (the "Eight Americas") defined on the basis of race and the location and socioeconomic characteristics of county of residence, in 2005. We combined data from the National Health and Nutrition Examination Survey and the Behavioral Risk Factor Surveillance System to estimate unbiased risk factor levels for the Eight Americas. We used data from the National Center for Health Statistics to estimate age-sex-disease-specific number of deaths in 2005. We used systematic reviews and meta-analyses of epidemiologic studies to obtain risk factor effect sizes for disease-specific mortality. We used epidemiologic methods for multiple risk factors to estimate the effects of current exposure to these risk factors on death rates, and life table methods to estimate effects on life expectancy. Asians had the lowest mean body mass index, fasting plasma glucose, and smoking; whites had the lowest systolic blood pressure (SBP). SBP was highest in blacks, especially in the rural South--5-7 mmHg higher than whites. The other three risk factors were highest in Western Native Americans, Southern low-income rural blacks, and/or low-income whites in Appalachia and the Mississippi Valley. Nationally, these four risk factors reduced life expectancy at birth in 2005 by an estimated 4.9 y in men and 4.1 y in women. Life expectancy effects were smallest in Asians (M, 4.1 y; F, 3.6 y) and largest in Southern rural blacks (M, 6.7 y; F, 5.7 y). Standard deviation of life expectancies in the Eight Americas would decline by 0.50 y (18%) in men and 0.45 y (21%) in women if these risks had been reduced to optimal levels. Disparities in the probabilities of dying from cardiovascular diseases and diabetes at different ages would decline by 69%-80%; the corresponding reduction for probabilities of dying from cancers would be 29%-50%. Individually, smoking and high blood pressure had the largest effect on life expectancy disparities. Disparities in smoking, blood pressure, blood glucose, and adiposity explain a significant proportion of disparities in mortality from cardiovascular diseases and cancers, and some of the life expectancy disparities in the US. Please see later in the article for the Editors' Summary.