Effect of physical training on skeletal muscle metabolism and ultrastructure in 70 to 75-year-old men.
The effects of a 12-week program of physical training on skeletal muscle (vastus lateralis) characteristics in 5 old men were investigated. Heart rates during submaximal bicycle exercise were decreased after training, indicating an improved cardiovascular function. As judged from enzyme activity measurements, the anaerobic capacity as well as the mitochondrial oxidative capacity were increased by the training. Fatty acid oxidation capacity remained unchanged, while the glycolytic potential tended to be increased, suggesting a somwehat different pattern of adaptation as compared to that seen in young subjects. Volume fractions of mitochondria and lipid droplets were unchanged with training, supporting the view that in old men, increases in oxidative capacity take place within the existing mitochondrial volume. It was concluded, that the aging human skeletal muscle remains trainable, and that the training response is similar, but possibly not identical, to that seen in younger age groups.


Biochemical and physical characterisation of urinary nanovesicles following CHAPS treatment.
Urinary exosomes represent a precious source of potential biomarkers for disease biology. Currently, the methods for vesicle isolation are severely restricted by the tendency of vesicle entrapment, e.g. by the abundant Tamm-Horsfall protein (THP) polymers. Treatment by reducing agents such as dithiothreitol (DTT) releases entrapped vesicles, thus increasing the final yield. However, this harsh treatment can cause remodelling of all those proteins which feature extra-vesicular domains stabilized by internal disulfide bridges and have detrimental effects on their biological activity. In order to optimize exosomal yield, we explore two vesicle treatment protocols - dithiothreitol (DTT) and 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonic (CHAPS) - applied to the differential centrifugation protocol for exosomal vesicle isolation. The results show that CHAPS treatment does not affect vesicle morphology or exosomal marker distribution, thus eliminating most of THP interference. Moreover, the recovery and preservation of catalytic activity of two trans-membrane proteases, dipeptidyl peptidase IV and nephrilysin, was examined and found to be clearly superior after CHAPS treatment compared to DTT. Finally, proteomic profiling by mass spectrometry (MS) revealed that 76.2% of proteins recovered by CHAPS are common to those seen for DTT treatment, which illustrates underlining similarities between the two approaches. In conclusion, we provide a major improvement to currently-utilized urinary vesicle isolation strategies to allow recovery of urinary vesicles without the deleterious interference of abundant urinary proteins, while preserving typical protein folding and, consequently, the precious biological activity of urinary proteins which serve as valuable biomarkers.


[Lipid composition of a Corynebacterium mediolanum culture transforming steroids].
Lipid composition was studied in Corynebacterium mediolanum during its growth in complex organic media favouring a high transformation activity. Lipid composition of the media was also analyzed. Lipid composition of the culture grown in a synthetic medium was comparatively studied. C. mediolanum was shown to have a peculiar lipid spectrum which did not depend on the growth medium composition and whose main component was an unidentified glycolipid (60-80% of the total lipid content). The lipids of the culture included also phosphatidyl glycerol, diphosphatidyl glycerol, a minor unidentified glycolipid, mono-, di- and triglycerides, free fatty acids, hydrocarbons and, in trace amounts, sterols and their esters. The fatty acids were represented mainly by anteiso-C15, iso-C16 and anteiso-C17. The total lipid content varied from 3 to 6% of the dry biomass weight depending on the growth phase. A correlation was established between a change in the lipid content of C. mediolanum (in particular, phosphatidyl glycerol) and its transformation activity. The lipids present in the media were found to inhibit the transformation of 21-acetate 3 beta,17 alpha,21-trihydroxy-5-pregnenone-20 (monoacetate of substance R, MAR) into the Reichstein substance S, 17 alpha,21-dihydroxy-4-pregnenedione-3,20 performed by the culture. The rate of MAR transformation by the growing culture rose when lipids were eliminated from the composition of organic media.


Isoflavone levels in soy foods consumed by multiethnic populations in Singapore and Hawaii.
Concentrations and glucosidic conjugation patterns of isoflavones were determined in soy foods consumed by multiethnic populations in Singapore and Hawaii. Six raw and 11 cooked food groups traditionally consumed in Singapore and 8 food groups consumed in Hawaii were analyzed by reversed-phase high-pressure liquid chromatography with diode array detection. Mean total isoflavone levels varied between 35 and 7500 ppm, with the lowest values found in soy milk and burgers and the highest levels observed in soybean and its seeds and in supplements. Total isoflavone levels and conjugation patterns varied as a function of soybean variety, storage conditions, and food processing. A large contribution to the differences in total isoflavone content between food groups was due to the water content in foods and to leaching of polar analytes into the water phase during boiling. Soy protein drinks and traditional soy foods were found to possess very similar isoflavone amounts considering usual serving sizes.


Severity of insomnia correlates with cognitive impairment.
Cognitive deficits in insomnia have been already reported (5), however, a correlation between cognitive impairment and severity of insomnia was not as yet studied. Sixteen not medicated patients with primary insomnia according to DSM-IV (4), 7 men an 9 women, of mean age 40.8 year, were compared to 16 controls, matched according to age, sex and education. Standard polysomnographic data (PSG) were recorded. The next day all the subjects completed Athens Insomnia Scale (AIS) (7), Hyperarousal Scale (HS) (6), Hamilton Depression Rating Scale (HAM-D), Beck Depression Inventory (BDI), Multiple Sleep Latency Test (MSLT) (3), Continuous Attention Test (CAT) with simple reaction time (RT) (8) and Selective Reminding Test (SRT) (2). The psychophysiological differences between patients and controls are shown in Table I. HAM-D and BDI scores were elevated in patients, although none of the patients met clinical criteria of depression. Insomniacs did not differ in the immediate recall, but the number of repetitions necessary to learn all the items of SRT was greater in patients. Insomniacs usually complain of poor performance, however, learning impairment has not been documented in insomnia. Degree of the learning impairment correlated with insomnia score (Fig. 1). Cognitive deficit cannot be due to a daytime sleepiness because sleep latency in all MSLT sessions was not shorter in insomniacs. No correlations between results of SRT and standard PSG parameters were found, in accordance with the thesis that subjective feeling of nonrestorative sleep and other accompanying deficits are only symptoms of an underlying 24-hour disorder (1).


Sickle cell disease patients' perceptions of emergency department pain management.
Patients with sickle cell disease (SCD) experience painful crises that often require admission to the emergency department (ED) for pain management. Factors such as ED overcrowding and negative perception and stigmatization of SCD may impact patients' perceptions of the quality of pain management in the ED. Data from a multisite prospective cohort study was assessed to determine whether demographic (age and sex), clinical (time to administration of initial analgesia, number of analgesic doses, discharge disposition, and clinical site), or interpersonal factors (separately measured perceptions of being treated with trust and respect by ED triage nurses, nurses, and physicians) were associated with patient ratings of their pain management in the ED. Patients were adults with SCD seen at 3 EDs (2 urban and 1 rural). Demographic and clinical information was derived from medical record review; interpersonal and ED pain management ratings were derived from interviews conducted 1 week post ED visit. A total of 209 interviews by 98 patients were analyzed. Results indicated significant differences among the ED sites on the demographic, clinical, and interpersonal factors. Overall, patients reported being treated with trust and respect by ED clinicians. Adjusted logistic regression analyses indicated that ED clinical site 1 (odds ratio [OR], 10.42; 95% confidence interval [Cl], 1.44-7.36) and being treated with trust and respect by the ED physician (OR, 25.53; 95% CI, 2.07-314.96) predicted good ED pain management ratings. Interpersonal health care experiences may be an important indicator of patient satisfaction and quality of care received by patients with SCD in the ED.


Deficiency of lncRNA SNHG12 impairs ischemic limb neovascularization by altering an endothelial cell cycle pathway.
SNHG12, a long noncoding RNA (lncRNA) dysregulated in atherosclerosis, is known to be a key regulator of vascular senescence in endothelial cells (ECs). However, its role in angiogenesis and peripheral artery disease has not been elucidated. Hind-limb ischemia studies using femoral artery ligation (FAL) in mice showed that SNHG12 expression falls readily in the acute phase of the response to limb ischemia in gastrocnemius muscle and recovers to normal when blood flow recovery is restored to ischemic muscle, indicating that it likely plays a role in the angiogenic response to ischemia. Gain- and loss-of-function studies demonstrated that SNHG12 regulated angiogenesis - SNHG12 deficiency reduced cell proliferation, migration, and endothelial sprouting, whereas overexpression promoted these angiogenic functions. We identified SNHG12 binding partners by proteomics that may contribute to its role in angiogenesis, including IGF-2 mRNA-binding protein 3 (IGF2BP3, also known as IMP3). RNA-Seq profiling of SNHG12-deficient ECs showed effects on angiogenesis pathways and identified a strong effect on cell cycle regulation, which may be modulated by IMP3. Knockdown of SNHG12 in mice undergoing FAL using injected gapmeRs) decreased angiogenesis, an effect that was more pronounced in a model of insulin-resistant db/db mice. RNA-Seq profiling of the EC and non-EC compartments in these mice revealed a likely role of SNHG12 knockdown on Wnt, Notch, and angiopoietin signaling pathways. Together, these findings indicate that SNHG12 plays an important role in the angiogenic EC response to ischemia.


Modelling neural correlates of working memory: a coordinate-based meta-analysis.
Working memory subsumes the capability to memorize, retrieve and utilize information for a limited period of time which is essential to many human behaviours. Moreover, impairments of working memory functions may be found in nearly all neurological and psychiatric diseases. To examine what brain regions are commonly and differently active during various working memory tasks, we performed a coordinate-based meta-analysis over 189 fMRI experiments on healthy subjects. The main effect yielded a widespread bilateral fronto-parietal network. Further meta-analyses revealed that several regions were sensitive to specific task components, e.g. Broca's region was selectively active during verbal tasks or ventral and dorsal premotor cortex were preferentially involved in memory for object identity and location, respectively. Moreover, the lateral prefrontal cortex showed a division in a rostral and a caudal part based on differential involvement in task set and load effects. Nevertheless, a consistent but more restricted "core" network emerged from conjunctions across analyses of specific task designs and contrasts. This "core" network appears to comprise the quintessence of regions, which are necessary during working memory tasks. It may be argued that the core regions form a distributed executive network with potentially generalized functions for focussing on competing representations in the brain. The present study demonstrates that meta-analyses are a powerful tool to integrate the data of functional imaging studies on a (broader) psychological construct, probing the consistency across various paradigms as well as the differential effects of different experimental implementations.


Saline as a vehicle control does not alter ventilation in male CD-1 mice.
Saline (0.9% NaCl) is used in clinical and research settings as a vehicle for intravenous drug administration. While saline is a standard control in mouse studies, there are reports of hyperchloremic metabolic acidosis in high doses. It remains unknown if metabolic acidosis occurs in mice and/or if compensatory increases in breathing frequency and tidal volume accompany saline administration. It was hypothesized that saline administration alters blood pH and the pattern of breathing in conscious CD-1 male mice exposed to air or hypoxia (10% O2 , balanced N2 ). Unrestrained barometric plethysmography was used to quantify breathing frequency (breaths/min; bpm), tidal volume (VT; mL/breath/10 g body weight (BW)), and minute ventilation (VE; mL/min/10 g BW) in two designs: (1) 11-week-old mice with no saline exposure (n = 11) compared to mice with 7 days of 0.9% saline administration (intraperitoneal, i.p.; 10 mL/kg body mass; n = 6). and (2) 17-week-old mice tested before (PRE) and after 1 day (POST1, n = 6) or 7 days (POST7, n = 5) of saline (i.p.; 10 mL/kg body mass). There were no differences when comparing frequency, VT, or VE between groups for either design with room air or hypoxia exposures. Hypoxia increased frequency, VT, and VE compared to room air. Moreover, conscious blood sampling showed no differences in pH, paCO2 , paO2 , or HCO3- in mice without or with 7 days of saline. These findings reveal no differences in ventilation following 1 and/or 7 days of saline administration in mice. Therefore, the use of 0.9% saline as a control is supported for studies evaluating the control of breathing in mice.


Continuous EEG monitoring detects nonconvulsive seizure and Ictal-Interictal Continuum abnormalities in moderate to severe ICANS following systemic CAR-T therapy.
Immune Cell Effector Associated Neurotoxicity Syndrome (ICANS) is common amongst patients receiving CD19 targeted Chimeric Antigen Receptor T-cell (CAR-T) therapy. The purpose of this study is to characterize the incidence of seizures and ictal-interictal continuum (IIC) abnormalities in patients with ICANS. Retrospective review of consecutive patients treated with axicabtagene ciloleucel (axi-cel) for recurrent high-grade systemic lymphoma at Stanford Medical Center between 2/2016-6/2019. Electronic medical records (EMR) were reviewed for clinical features, treatment information, EEG data, CRS (cytokine release syndrome)/ICANS severity, and clinical outcomes. Fifty-six patients met inclusion criteria. 85.7% of patients developed CRS, and 58.9% developed ICANS. Twenty-eight patients had EEG monitoring, of whom 26 had ICANS. Median duration of EEG monitoring was 30 hours (range .5-126 hours). Four patients (7.1%) had seizures (1 patient had a clinical generalized seizure, 2 patients had clinical and nonconvulsive seizures, and 1 patient had an isolated non-convulsive seizure). Ictal-interictal continuum abnormalities were common, of which generalized periodic discharges (GPDs) with triphasic morphology and GPDs with epileptiform morphology were most frequently seen. Generalized periodic discharges with triphasic wave morphology were found across Grade 2-3 peak ICANS severity, however the majority (86%) of patients with epileptiform GPDs had Grade 3 peak ICANS severity. Among patients receiving axi-cel, seizure occurred in 7.1% of the total cohort, representing 12% of patients with ICANS. Ictal-interictal continuum abnormalities are also seen in patients with ICANS, most commonly GPDs. 75% of patients with seizures had nonconvulsive seizures supporting the use of continuous video EEG monitoring in this population.


Contrasting patterns of nucleotide polymorphism suggest different selective regimes within different parts of the PgiC1 gene in Festuca ovina L.
Phosphoglucose isomerase (PGI, EC 18.104.22.168) is an essential metabolic enzyme in all eukaryotes. An earlier study of the PgiC1 gene, which encodes cytosolic PGI in the grass Festuca ovina L., revealed a marked difference in the levels of nucleotide polymorphism between the 5' and 3' portions of the gene. In the present study, we characterized the sequence polymorphism in F. ovina PgiC1 in more detail and examined possible explanations for the non-uniform pattern of nucleotide polymorphism across the gene. Our study confirms that the two portions of the PgiC1 gene show substantially different levels of DNA polymorphism and also suggests that the peptide encoded by the 3' portion of PgiC1 is functionally and structurally more important than that encoded by the 5' portion. Although there was some evidence of purifying selection (dN/dS test) on the 5' portion of the gene, the signature of purifying selection was considerably stronger on the 3' portion of the gene (dN/dS and McDonald-Kreitman tests). Several tests support the action of balancing selection within the 5' portion of the gene. Wall's B and Q tests were significant only for the 5' portion of the gene. There were also marked peaks of nucleotide diversity, Tajima's D and the dN/dS ratio at or around a PgiC1 codon site (within the 5' portion of the gene) that a previous study had suggested was subject to positive diversifying selection. Our results suggest that the two portions of the gene have been subject to different selective regimes. Purifying selection appears to have been the main force contributing to the relatively low level of polymorphism within the 3' portion of the sequence. In contrast, it is possible that balancing selection has contributed to the maintenance of the polymorphism within the 5' portion of the gene.


Multiple-ligament knee injuries: a systematic review of the timing of operative intervention and postoperative rehabilitation.
Traumatic knee dislocations that result in multiple-ligament knee injuries are unusual and are poorly studied. We are not aware of any prospective data regarding their treatment. Both the optimum timing of surgery for repair or reconstruction and the aggressiveness of rehabilitation are debated. The purpose of this systematic review was to compare the outcomes of early, delayed, and staged procedures as well as the subsequent rehabilitation protocols. We surveyed the literature and retrieved twenty-four retrospective studies, involving 396 knees, dealing with the surgical treatment of the most severe multiple-ligament knee injuries (those involving both cruciate ligaments and either or both collateral ligaments). Data were extracted, and surgical timing was categorized as acute, chronic, or staged. Early postoperative mobility and immobilization were also compared. We found that acute treatment was associated with residual anterior knee instability when compared with chronic treatment (odds ratio, 2.58; 95% confidence interval, 1.2 to 5.8; p = 0.018). Significantly more patients who were managed acutely were found to have more flexion deficits when compared with those who were managed chronically (odds ratio, 5.18; 95% confidence interval, 1.5 to 17.5; p = 0.004). Staged treatments yielded the highest percentage of excellent and good subjective outcomes (79%; 95% confidence interval, 62.2% to 89.3%). Additional treatment for joint stiffness was significantly more likely in association with acute treatment (17%; 95% confidence interval, 13.0% to 22.4%; p < 0.001) and staged treatment (15%; 95% confidence interval, 7.6% to 28.2%; p = 0.001) when each was compared with chronic treatment (0% [zero of seventy-one]; 95% confidence interval, 0.0% to 5.1%). Early mobility was not associated with increased joint instability in acutely managed patients. Early mobility yielded fewer range-of-motion deficits but did not reduce the rate of follow-up manipulation or arthrolysis. This review of the available literature suggests that delayed reconstructions of severe multiple-ligament knee injuries could potentially yield equivalent outcomes in terms of stability when compared with acute surgery. However, in the acutely managed patient, early mobility is associated with better outcomes in comparison with immobilization. Acute surgery is highly associated with range-of-motion deficits. Staged procedures may produce better subjective outcomes and a lower number of range-of-motion deficits but are still likely to require additional treatment for joint stiffness. More aggressive rehabilitation may prevent this from occurring in multiple-ligament knee injuries that are treated acutely.


Fundamental questions and concepts about photoreception and the case of Euglena gracilis.
The ability to sense light can be considered the most fundamental and presumably the most ancient property of visual systems. This ability is the basis of phototaxis, one of the most striking behavioral responses of motile photosynthetic microorganisms (i.e. microalgae) to light stimuli, which allows them to move toward or away directional light. In order to fully exploit the information content of light (intensity, direction, distribution) microorganisms need proper perceiving devices, termed photoreceptors, which must act as sensors, to perceive wavelength and direction of light, as transducers, to convert the light signal into chemical and/or electrical information, but also as amplifiers and eventually as transmitters. This review describes the universal structural, behavioral and physiological features necessary for the proper functioning of these devices in algae, and how these features have been investigated by means of different analytical techniques such as for example microspectroscopy, digital fluorescence microscopy, two photons FLIM. The insight of the photoreceptive response mechanism is explained using the unicellular alga Euglena gracilis, in which the different structural, behavioral and physiological features combine to achieve a concerted, efficient response to light stimuli.


Phosphorylation of PSII proteins in maize thylakoids in the presence of Pb ions.
Lead is potentially toxic to all organisms including plants. Many physiological studies suggest that plants have developed various mechanisms to contend with heavy metals, however the molecular mechanisms remain unclear. We studied maize plants in which lead was introduced into detached leaves through the transpiration stream. The photochemical efficiency of PSII, measured as an Fv/Fm ratio, in the maize leaves treated with Pb was only 10% lower than in control leaves. The PSII activity was not affected by Pb ions in mesophyll thylakoids, whereas in bundle sheath it was reduced. Protein phosphorylation in mesophyll and bundle sheath thylakoids was analyzed using mass spectrometry and protein blotting before and after lead treatment. Both methods clearly demonstrated increase in phosphorylation of the PSII proteins upon treatment with Pb(2+), however, the extent of D1, D2 and CP43 phosphorylation in the mesophyll chloroplasts was clearly higher than in bundle sheath cells. We found that in the presence of Pb ions there was no detectable dephosphorylation of the strongly phosphorylated D1 and PsbH proteins of PSII complex in darkness or under far red light. These results suggest that Pb(2+) stimulates phosphorylation of PSII core proteins, which can affect stability of the PSII complexes and the rate of D1 protein degradation. Increased phosphorylation of the PSII core proteins induced by Pb ions may be a crucial protection mechanism stabilizing optimal composition of the PSII complexes under metal stress conditions. Our results show that acclimation to Pb ions was achieved in both types of maize chloroplasts in the same way. However, these processes are obviously more complex because of different metabolic status in mesophyll and bundle sheath chloroplasts.


Quantitative 3D analysis of complex single border cell behaviors in coordinated collective cell migration.
Understanding the mechanisms of collective cell migration is crucial for cancer metastasis, wound healing and many developmental processes. Imaging a migrating cluster in vivo is feasible, but the quantification of individual cell behaviours remains challenging. We have developed an image analysis toolkit, CCMToolKit, to quantify the Drosophila border cell system. In addition to chaotic motion, previous studies reported that the migrating cells are able to migrate in a highly coordinated pattern. We quantify the rotating and running migration modes in 3D while also observing a range of intermediate behaviours. Running mode is driven by cluster external protrusions. Rotating mode is associated with cluster internal cell extensions that could not be easily characterized. Although the cluster moves slower while rotating, individual cells retain their mobility and are in fact slightly more active than in running mode. We also show that individual cells may exchange positions during migration.


Sarcomere length control in striated muscle.
A system that makes control of muscle length (ML), sarcomere length (SL), and force (F) possible in striated muscle preparations is described. SL was measured by light diffraction techniques and two diffractometers. Control was performed by influencing ML with a penmotor system with a frequency response of 190 Hz. SL or F could be controlled by interrupting the internal position (i.e., ML) feedback of the motor and by closing the respective loop. Velocity feedback of the motor through an internal velocity coil was maintained in all cases for optimal damping. Steady-state error of the system was minimized by an integrating loop filter. The feedback path was selected by means of potentiometers or analog switches. Electronic stops in the circuit protected the muscle against excessive stretch and load. A microprocessor-based average-response computer could be used for feedforward control to eliminate noise or to analyze longitudinal uniformity of the muscle. Responses of rat cardiac trabeculae during SL and F control are shown. Transient behavior of SL and F during control and measures to eliminate the transients are discussed.


Sexual behaviours and sexually transmitted infection outcomes in a cohort of HIV-negative men who have sex with men attending sexual health clinics in England.
Men who have sex with men (MSM) attending sexual health (SH) clinics are at high risk for HIV acquisition and are disproportionately affected by sexually transmitted infections (STIs). We collected standardised behavioural data from MSM attending clinics to characterise sexual behaviours and identify predictors for HIV and STIs. In 2012–2013, HIV-negative MSM attending five SH clinics in England reported sexual behaviours in the previous three months via a self-administered questionnaire. Behaviours were linked to the individual's clinical records using national surveillance. The prevalence and incidence of bacterial STIs (gonorrhoea, Chlamydia, lymphogranuloma venereum and syphilis) and incidence of HIV were calculated. Adjusted odds ratios and hazard ratios with 95% confidence interval (CI) were reported for significant predictors. Of 1278 HIV-negative MSM, 54% were of white ethnicity and UK-born and 43% were 25–34 years old. Almost all men reported at least one partner in the last three months. Half reported condomless anal sex and 36% condomless receptive anal intercourse (CRAI). Incidence of bacterial STIs was 46/100 (95%CI 39–54) person years (py) and of HIV was 3.1/100 (95%CI 1.7–5.6) py. A STI at baseline and CRAI with increasing numbers of partners were associated with both incident infections. In this cohort of MSM high-risk behaviours and STIs were prevalent. Engagement in CRAI increased the likelihood of subsequent infection, while men diagnosed with a bacterial STI were at increased risk of a future STI. Clinical and behavioural risk assessments to determine an individual's risk of infection could allow a more nuanced prevention approach that has greater success in reducing transmission.


What is the public appetite for healthy eating policies? Evidence from a cross-European survey.
World Health Organization estimates that obesity accounts for 2-8% of health care costs in different parts of Europe, and highlights a key role for national policymaking in curbing the epidemic. A variety of healthy-eating policy instruments are available, ranging from more paternalistic policies to those less intrusive. Our aim is to measure and explain the level of public support for different types of healthy eating policy in Europe, based on data from a probabilistic sample of 3003 respondents in five European countries. We find that the main drivers of policy support are attitudinal factors, especially attribution of obesity to excessive availability of unhealthy foods, while socio-demographic characteristics and political preferences have little explanatory power. A high level of support for healthy eating policy does not translate into acceptance of higher taxes to fund them, however.


Modeling sound transmission through the pulmonary system and chest with application to diagnosis of a collapsed lung.
A theoretical and experimental study was undertaken to examine the feasibility of using audible-frequency vibro-acoustic waves for diagnosis of pneumothorax, a collapsed lung. The hypothesis was that the acoustic response of the chest to external excitation would change with this condition. In experimental canine studies, external acoustic energy was introduced into the trachea via an endotracheal tube. For the control (nonpneumothorax) state, it is hypothesized that sound waves primarily travel through the airways, couple to the lung parenchyma, and then are transmitted directly to the chest wall. In contradistinction, when a pneumothorax is present the intervening air presents an added barrier to efficient acoustic energy transfer. Theoretical models of sound transmission through the pulmonary system and chest region to the chest wall surface are developed to more clearly understand the mechanisms of intensity loss when a pneumothorax is present, relative to a baseline case. These models predict significant decreases in acoustic transmission strength when a pneumothorax is present, in qualitative agreement with experimental measurements. Development of the models, their extension via finite element analysis, and comparisons with experimental canine studies are reviewed.


Emergence of new pandemic GII.4 Sydney norovirus strain correlates with escape from herd immunity.
GII.4 noroviruses are a significant source of acute gastroenteritis worldwide, causing the majority of human norovirus outbreaks. Evolution of the GII.4 major capsid protein occurs rapidly, resulting in the emergence of new strains that produce successive waves of pandemic disease. A new pandemic isolate, GII.4 2012 Sydney, largely replaced previously circulating strains in late 2012. We compare the antigenic properties of GII.4 2012 Sydney with previously circulating strains. To determine whether GII.4-2012 Sydney is antigenically different from recently circulating strains GII.4-2006 Minerva and GII.4-2009 New Orleans in previously identified blockade epitopes, we compared reactivity and blockade profiles of GII.4-2006, GII.4-2009, and GII.4-2012 virus-like particles in surrogate neutralization/blockade assays using monoclonal antibodies and human polyclonal sera. Using monoclonal antibodies that map to known blockade epitopes in GII.4-2006 and GII.4-2009 and human outbreak polyclonal sera, we demonstrate either complete loss or significantly reduced reactivity and blockade of GII.4.2012 compared to GII.4-2006 and GII.4-2009. GII.4-2012 Sydney is antigenically different from GII.4-2006 Minerva and GII.4-2009 New Orleans in at least 2 key blockade epitopes. Viral evolution in key potential neutralization epitopes likely allowed GII.4-2012 to escape from human herd immunity and emerge as the new predominant strain.


Toxicity potential of absorbed-retained ethylene oxide residues in culture dishes on embryo development in vitro.
Four hundred eight-cell mouse embryos were cultured in vitro in either polystyrene (plastic) dishes (Exp. 1) or watch glasses (Exp. 2) to analyze the toxicity potential of absorbed-retained ethylene oxide (EtO). Culture dishes were gas-sterilized with Anprolene equipment and allowed to aerate (21 C) for varying durations. Post-sterilization EtO residues, as determined by weight measurement, were eluted from polystyrene dishes at an exponential rate. After 1 wk of aeration, .625 mg EtO was retained/g of polystyrene product. To determine the effect of EtO residue on in vitro culture, embryos were collected from superovulated donor mice (C57BL/6N), pooled in a modified Dulbecco's phosphate buffered saline (PBI) medium and then randomly allotted to dishes subjected to various aeration durations. Embryos were cultured in Whitten's medium +3 mg/ml bovine serum albumin (BSA) under a humidified 5% CO2 in air atmosphere at 37 C. Gross assessments of embryo development were made using standard morphology and quality grading systems and a fluorescein diacetate staining assay. In Exp. 1, embryo development was retarded at the 8- to 16-cell stage when less than or equal to 12 h aeration of polystyrene dishes was permitted. Aeration for 24 and 36 h resulted in suboptimal embryo development with fewer (P less than .01) blastocysts and greater degeneration rates at 48 h of in vitro culture compared with the control (manufacturer-packaged dishes, no EtO) and 1-wk aeration treatment groups.(ABSTRACT TRUNCATED AT 250 WORDS)


Comparison of the fecal microbiota of two monogastric herbivorous and five omnivorous mammals.
Fecal microbiota in seven different monogastric animal species, elephant, horse, human, marmoset, mouse, pig and, rat were compared using the same analytical protocol of 16S rRNA metagenome. Fecal microbiota in herbivores showed higher alpha diversity than omnivores except for pigs. Additionally, principal coordinate analysis based on weighted UniFrac distance demonstrated that herbivores and pigs clustered together, whereas other animal species were separately aggregated. In view of butyrate- and lactate-producing bacteria, predominant genera were different depending on animal species. For example, the abundance of Faecalibacterium, a known butyrate producer, was 8.02% ± 3.22% in human while it was less than 1% in other animal species. Additionally, Bifidobacterium was a predominant lactate producer in human and marmoset, while it was rarely detected in other omnivores. The abundance of lactate-producing bacteria in herbivores was notably lower than omnivores. On the other hand, herbivores as well as pig possess Fibrobacter, a cellulolytic bacterium. This study demonstrated that fecal microbiota in herbivorous animals is similar, sharing some common features such as higher alpha diversity and higher abundance of cellulolytic bacterium. On the other hand, omnivorous animals seem to possess unique fecal microbiota. It is of interest that pigs, although omnivore, have fecal microbiota showing some common features with herbivores.


Does HIV infection modify the relationship between pre-pregnancy body mass index and adverse birth outcomes?
South Africa faces dual epidemics of HIV and obesity; however, little research has explored whether HIV status influences associations between pre-pregnancy body mass index (BMI) and adverse birth outcomes. To examine associations between pre-pregnancy body mass index (BMI) and adverse birth outcomes, and if they differ by HIV status. We followed HIV-uninfected and -infected pregnant women initiating antiretroviral therapy (ART) from first antenatal visit through delivery. HIV-infected women initiated ART (tenofovir-emtricitabine/lamivudine-efavirenz) in pregnancy. Estimated pre-pregnancy BMI (kg/m2 ) was categorised as underweight (<18.5), normal (18.5-24.9), overweight (25.0-29.9), and obese (≥30.0). We used modified Poisson regression to estimate risk ratios (RR) for associations between pre-pregnancy BMI and adverse birth outcomes and explored modification by HIV status. Among 1116 women (53% HIV-infected), 44% of HIV-uninfected women and 36% of HIV-infected women were classified as obese; 4% of women were underweight. Overall, 12% of infants were delivered preterm (<37 weeks), 10% small for gestational age (SGA, <10th percentile), and 9% large for gestational age (LGA, >90th percentile). Compared to HIV-uninfected women, HIV-infected women on ART had less LGA (5% vs 13%) but more SGA (12% vs 8%), and a similar proportion of preterm (13% vs 11%) infants. Pre-pregnancy BMI was not associated with preterm birth. Among HIV-uninfected women, obesity modestly increased the risk of LGA (RR 1.34, 95% confidence interval [CI] 0.82, 2.19), and underweight modestly elevated the risk of SGA (RR 1.66, 95% CI 0.79, 3.46). These associations were attenuated among HIV-infected women (RR 1.07, 95% CI 0.44, 2.64 for LGA, and RR 1.34, 95% CI 0.49, 3.64 for SGA). In this urban African setting of high HIV prevalence, pre-pregnancy obesity was common and did not vary by HIV status. In HIV-uninfected women, obesity increased the risk of LGA and being underweight the risk of SGA, compared with among HIV-uninfected women.


Precision medicines for B-cell leukaemias and lymphomas; progress and potential pitfalls.
There is now a plethora of new precision medicines for B-cell malignancy including 'classical' kinase inhibitors, rationally designed inhibitors of anti-apoptotic proteins and antibody or antibody drug/toxin conjugates with functional properties. Some are showing spectacular single agent activity in early phase clinical studies and may reduce or, in combination, even obviate the need for chemotherapy. Nevertheless, significant problems remain if these medicines are to be introduced into routine clinical practice in a rational and affordable manner. Firstly, precision medicines must be carefully matched in a mechanistic fashion with specific subtypes of disease. Whilst sensitivity may be predicted by the detection of key mutations or by expression of target molecules, for therapies that depend on intact intracellular signalling pathways, functional assessment on viable primary malignant cells will be necessary using assays that faithfully mimic in vivo conditions. A second, but no less important challenge is to define mechanism-based synergistic combinations associated with minimal toxicities rather than simply adding new precision medicines to existing chemotherapeutic regimens. Finally, a closer, open, two-way interaction between academic medicine and the pharmaceutical industry will be necessary to achieve these aims. Implementing such changes would change radically how and where patients with B-cell malignancies are managed.


Some observations on the kindling process.
Kindling is an animal model of epilepsy in which repeated administration of a subconvulsant stimulus, electrical or chemical, produces a gradually increasing electroencephalographic and behavioral response which culminates in a behavioral seizure. The biological basis of the kindling effect remains unknown but alterations in neurotransmission figure prominently in most hypotheses. Several factors of kindling are considered as central to the phenomenon. It is most important to recognize that the development of kindling and a kindled seizure are not necessarily the same phenomenon. Kindling is essentially a permanent change in the sensitivity of the brain to a stimulus. It therefore follows that the biological basis for kindling must be a permanent change. Permanent changes in neurotransmitter levels or receptor parameters have not been conclusively demonstrated in kindling. Other possible permanent changes include changes in Ca++ activated mechanisms which alter neuronal structures such as dendritic spines. An essential component to the riddle of kindling is the absolute requirement for an inter-stimulus interval of at least 1-2 hours. This suggests that the biological process which leads to kindling occurs in this critical period. Recent experiments with cysteamine suggest that the events in this critical period can be manipulated chemically. An understanding of these events will help to clarify the biological basis of kindling.


CMV-encoded Fcγ receptors: modulators at the interface of innate and adaptive immunity.
The constant region of IgG antibodies mediates antiviral activities upon engaging host Fcγ receptors (FcγRs) expressed by a variety of immune cells, such as antibody-dependent cellullar cytotoxcity (ADCC) executed by natural killer (NK)cells. Human cytomegalovirus (HCMV) is unique among viruses by encoding also an array of several Fcγ-binding glycoproteins with cell surface disposition and concomitant incorporation into the virion. Evidence is increasing that the virus-encoded Fcγ receptors differ in their Fcγ binding mode but effectively operate as adversaries of host FcγRs since they are able to prevent IgG-mediated triggering of activating host FcγRs, i.e., FcγRI, FcγRIIA, and FcγRIIIA. Here we discuss virus-encoded FcγRs as the first known HCMV inhibitors of IgG-mediated immunity which could account for the limited efficacy of HCMV hyperimmune globulin in clinical settings. A better understanding of their molecular mode of action opens up new perspectives for improving IgG therapies against HCMV disease.


A case of cutaneous ulcerative alternariosis: rare association with diabetes mellitus and unusual failure of itraconazole treatment.
Alternaria species are ubiquitous dematiaceous fungi that are increasingly recognized as pathogens in immunocompromised patients or those with significant underlying disease, but they are also pathogens in otherwise healthy hosts. We describe a case of dermal cutaneous ulcerative alternariosis in a frail 83-year-old patient with diet-controlled diabetes mellitus. Histological analysis revealed hyphal morphology in tissue sections that was initially confused with that of a zygomycetous fungus, and multiple positive culture results were necessary to identify the organism. Treatment with oral itraconazole and surgical debridement were ineffective; clinical improvement was achieved by means of treatment with intravenous amphotericin B lipid complex. We review the literature regarding the role of diabetes mellitus in cutaneous alternariosis and regarding the efficacy of treatment with itraconazole, which has been used very successfully. To our knowledge, this is only the second case report noting diabetes mellitus uncomplicated by steroid administration as a possible predisposing factor for cutaneous infection.


Validation of the CO2 rebreathing method for measuring cardiac output in patients with hypertension or heart failure.
The CO2 rebreathing method, a noninvasive indirect Fick procedure, has been previously validated in normal subjects at supine rest and during exercise. In the present study 29 nearly simultaneous measurements of cardiac output were made by CO2 rebreathing and dye dilution in 17 patients with cardiovascular disease, 11 with hypertension, and six with congestive heart failure. Cardiac output at supine rest averaged 5.52 +/- 0.31 L. per minute by CO2 and 5.62 +/- 0.32 L. per minute by dye dilution (r = 0.93). Successive cardiac output measurements by rebreathing varied 6.0 +/- 1.1 per cent (r = 0.96) and by dye dilution, 6.5 +/- 0.9 per cent. Changes in cardiac output during exercise or after beta-adrenergic blockade were reflected by the CO2 method. It is concluded that the CO2 method provides reliable measurements of cardiac output both at rest and during exercise in patients with hypertension or congestive heart failure. Further validation of the method is needed in various disease states before it can be widely applied.


Cryo-EM analyses reveal the common mechanism and diversification in the activation of RET by different ligands.
RET is a receptor tyrosine kinase (RTK) that plays essential roles in development and has been implicated in several human diseases. Different from most of RTKs, RET requires not only its cognate ligands but also co-receptors for activation, the mechanisms of which remain unclear due to lack of high-resolution structures of the ligand/co-receptor/receptor complexes. Here, we report cryo-EM structures of the extracellular region ternary complexes of GDF15/GFRAL/RET, GDNF/GFRα1/RET, NRTN/GFRα2/RET and ARTN/GFRα3/RET. These structures reveal that all the four ligand/co-receptor pairs, while using different atomic interactions, induce a specific dimerization mode of RET that is poised to bring the two kinase domains into close proximity for cross-phosphorylation. The NRTN/GFRα2/RET dimeric complex further pack into a tetrameric assembly, which is shown by our cell-based assays to regulate the endocytosis of RET. Our analyses therefore reveal both the common mechanism and diversification in the activation of RET by different ligands.


Road safety perspectives among employees of a multinational corporation in urban India: local context for global injury prevention.
In rapidly developing economies, like urban India, where road traffic injury rates are among the world's highest, the corporate workplace offers a non-traditional venue for road safety interventions. In partnership with a major multinational corporation (MNC) with a large Indian workforce, this study aimed to elicit local employee perspectives on road safety to inform a global corporate health platform. The safety attitudes and behaviours of 75 employees were collected through self-report survey and focus groups in the MNC offices in Bangalore and Pune. Analysis of these data uncovered incongruity between employee knowledge of safety strategies and their enacted safety behaviours and identified local preference for interventions and policy-level actions. The methods modelled by this study offer a straightforward approach for eliciting employee perspective for local road safety interventions that fit within a global strategy to improve employee health. Study findings suggest that MNCs can employ a range of strategies to improve the road traffic safety of their employees in settings like urban India including: implementing corporate traffic safety policy, making local infrastructure changes to improve road and traffic conditions, advocating for road safety with government partners and providing employees with education and access to safety equipment and safe transportation options.


Effects of chronic prenatal ethanol exposure on mitochondrial glutathione and 8-iso-prostaglandin F2alpha concentrations in the hippocampus of the perinatal guinea pig.
It is hypothesised that oxidative stress is a key mechanism of ethanol neurobehavioural teratogenicity, resulting in altered endogenous antioxidant status and increased membrane lipid peroxidation in the hippocampus of chronic prenatal ethanol exposure (CPEE) offspring. To test this hypothesis, timed pregnant guinea-pigs (term, approximately gestational day (GD) 68) received chronic daily oral administration of (i) 4 g ethanol kg(-1) maternal bodyweight, (ii) isocaloric sucrose with pair feeding, or (iii) water. At GD 65 (term fetus) and postnatal day (PD) 0 (neonate), individual offspring were killed, the brain was excised and the hippocampi were dissected. Glutathione (GSH) concentration was measured in the cytosolic and mitochondrial fractions of hippocampal homogenate. The occurrence of lipid peroxidation was determined by measuring the concentration of 8-iso-prostaglandin F2+/- (8-iso-PGF2+/-). There was CPEE-induced decreased brain weight and hippocampal weight at GD 65 and PD 0, decreased mitochondrial GSH concentration in the hippocampus at PD 0, with no change in mitochondrial GSH concentration at GD 65 or cytosolic GSH concentration at GD 65 or PD 0, and no change in mitochondrial or whole-homogenate 8-iso-PGF2+/- concentration in the hippocampus at GD 65 or PD 0. The data demonstrate that CPEE produces selective mitochondrial dysfunction in the hippocampus of the neonatal guinea-pig, involving GSH depletion.


Sexually dimorphic characters of the ultimate legs in lithobiid centipedes (Myriapoda, Chilopoda, and Lithobiomorpha): Morphology and implications for reproductive behavior.
Many species of lithobiomorph centipedes present a pronounced sexual dimorphism reflected in remarkable structural modifications on the ultimate legs of males. Most records of these male secondary sexual characters addressed taxonomy, helping to identify and characterize species or diagnose genera, but information on their diversity, detailed morphology and possible function(s) is scarce. In this study, nine species of the two lithobiid genera Lithobius Leach, 1814 and Eupolybothrus Verhoeff, 1907 were investigated, using light and scanning electron microscopy to document the detailed morphology of secondary sexual characters of male ultimate legs. Secondary sexual characters affecting the cuticle of the ultimate legs are described in detail and found to often be associated with sensilla, interpreted here as chemo- and mechanoreceptors, and with clusters of pores, a hitherto undescribed pore-distribution for this group. The tibial nodule of the species Lithobius nodulipes Latzel, 1880, was additionally examined with histological semi-thin sections. These results revealed that the clustered pores are connected to glandular tissue, and are, based on their morphology, interpreted as openings of flexo-canal epidermal glands. The presence of various sensory and glandular structures associated with sexual dimorphism indicates a likely role during courtship and mating. The closely related species examined in this research show comparable dimorphic structures, which are otherwise species-specific. Morphological observations on secondary sexual structures inform on reproductive biology in groups like lithobiomorphs for which there are limited behavioral data.


A comparative study of extracellular vesicle-associated and cell-free DNA and RNA for HPV detection in oropharyngeal squamous cell carcinoma.
This study compares the detection sensitivity of two separate liquid biopsy sources, cell-free (cf) DNA/RNA and extracellular vesicle (EV)-associated DNA/RNA (EV-DNA/RNA), to identify circulating Human Papilloma Virus (HPV) DNA/RNA in plasma obtained from patients with oropharyngeal squamous cell carcinoma (OPCSCC). We also report on the longitudinal changes observed in HPV-DNA levels in response to treatment. A prospective study was conducted that included 22 patients with locally advanced disease and six patients with metastatic OPCSCC. Twenty-three patients had HPV-related OPCSCC defined by p16 immunohistochemistry. Levels of circulating HPV-DNA and HPV-RNA from plasma-derived cf-DNA/RNA and EV-DNA/RNA were quantified using digital droplet PCR. Circulating HPV-DNA was detected with higher sensitivity in cf-DNA compared to EV-DNA at 91% vs. 42% (p = <0.001). Similarly, circulating tumoral HPV-RNA was detected at a higher sensitivity in cf-RNA compared to EV-RNA, at 83% vs. 50% (p = 0.0019). In the locally advanced cohort, 100% (n = 16) of HPV-OPCSCC patients demonstrated a reduction in circulating HPV-DNA levels in cf-DNA following curative treatment, with 81% of patients demonstrating complete clearance to undetectable levels. However, in metastatic HPV-OPCSCC patients (n = 4), HPV-DNA levels did not correlate with treatment response. Our study demonstrates that although HPV-DNA/RNA can be detected in EV associated DNA/RNA, cf-DNA/RNA is the more sensitive liquid biopsy medium. As circulating HPV-DNA levels were found to only correlate with treatment response in the locally advanced but not metastatic setting in our small cohort of patients, the use of HPV-DNA as a dynamic biomarker to monitor treatment response requires further evaluation.


The synthesis of fatty acids in avocado mesocarp and cauliflower bud tissue.
1. Plastid and mitochondrial preparations were obtained by density-gradient centrifugation of homogenates made by gentle disintergration of avocado fruit mesocarp and cauliflower bud tissue. 2. The mitochondrial preparations had respiratory activity but did not incorporate [1-14C]acetate into fatty acids. 3. The plastid preparations incorporated [1--14C]acetate into the range of fatty acids found in the parent tissue. No fatty acid synthetase activity could be detected in the 12000g supernatant of these homogenates. 4. Homogenates produced by rupture of the tissue in an Ato-Mix blender and plastid preparations disintegrated by ultrasonic treatment both had fatty acid synthetase activity which did not sediment at 105000g and which formed mainly [14-C]stearate from [2-14C]malonyl-CoA. 5. It is concluded that the plastids are the principal site of fatty acid biosynthesis in the tissues studied.


catena-Poly[[aqua-bis-(3-chloro-benzoato-κ(2) O,O')cadmium]-μ-N,N-di-ethyl-nico-tin-amide-κ(2) N (1):O].
In the crystal of the title Cd(II) polymeric complex, [Cd(C7H4ClO2)2(C10H14N2O)(H2O)] n , the Cd(II) cation is chelated by two chloro-benzoate anions and coordinated by two N,N-di--ethyl-nicotinamide (DENA) ligands and one water mol-ecule in a distorted NO6 penta-gonal-bipyramidal geometry. The Cd(II) cations are bridged by the pyridine N atom and carbonyl O atom of the DENA ligand to form a polymeric chain running along the b axis. Inter-molecular O-H⋯O hydrogen bonds between coordinating water mol-ecules and carboxyl-ate groups link adjacent chains into layers parallel to the bc plane. π-π contacts between benzene rings [shortest centroid-centroid distance = 3.912 (2) Å] further stabilizes the crystal structure. In the mol-ecule, weak C-H⋯O hydrogen bonds occur between the pyridine ring and carboxyl-ate groups; the dihedral angles between the carboxyl-ate groups and adjacent benzene rings are 4.6 (3) and 12.8 (3)°, while the benzene rings are oriented at a dihedral angle of 1.89 (13)°.


Visualization of endogenous glycine in cat retina: an immunocytochemical study with Fab fragments.
Fab fragments of a glycine antiserum were prepared and used for immunocytochemical visualization of glycine in the cat retina. The use of Fab fragments in conjunction with Fab-specific secondary and tertiary antisera improved tissue penetration and made it possible to identify a number of the immunoreactive neurons. Staining was observed in several subpopulations of amacrine cells, including the A7(AII) rod amacrine. Multiple subpopulations of cone bipolar cells were also seen to be immunoreactive. Many neurons exhibited no detectable immunostaining, indicating that general metabolic levels of glycine do not interfere with the visualization of those cells that contain large amounts of endogenous glycine. The distribution of immunostaining appears to parallel the pattern of glycine labeling seen previously with autoradiographic techniques and implicates these cells in glycine-mediated neurotransmission.


Photosynthetic biomass and H2 production by green algae: from bioengineering to bioreactor scale-up.
The development of clean borderless fuels is of vital importance to human and environmental health and global prosperity. Currently, fuels make up approximately 67% of the global energy market (total market = 15 TW year(-1)) (Hoffert et al. 1998). In contrast, global electricity demand accounts for only 33% (Hoffert et al. 1998). Yet, despite the importance of fuels, almost all CO(2) free energy production systems under development are designed to drive electricity generation (e.g. clean-coal technology, nuclear, photovoltaic, wind, geothermal, wave and hydroelectric). In contrast, and indeed almost uniquely, biofuels also target the much larger fuel market and so in the future will play an increasingly important role in maintaining energy security (Lal 2005). Currently, the main biofuels that are at varying stages of development include bio-ethanol, liquid carbohydrates [e.g. biodiesel or biomass to liquid (BTL) products], biomethane and bio-H(2). This review is focused on placing bio-H(2) production processes into the context of the current biofuels market and summarizing advances made both at the level of bioengineering and bioreactor design.


Whole-genome sequencing reveals oncogenic mutations in mycosis fungoides.
The pathogenesis of mycosis fungoides (MF), the most common cutaneous T-cell lymphoma (CTCL), is unknown. Although genetic alterations have been identified, none are considered consistently causative in MF. To identify potential drivers of MF, we performed whole-genome sequencing of MF tumors and matched normal skin. Targeted ultra-deep sequencing of MF samples and exome sequencing of CTCL cell lines were also performed. Multiple mutations were identified that affected the same pathways, including epigenetic, cell-fate regulation, and cytokine signaling, in MF tumors and CTCL cell lines. Specifically, interleukin-2 signaling pathway mutations, including activating Janus kinase 3 (JAK3) mutations, were detected. Treatment with a JAK3 inhibitor significantly reduced CTCL cell survival. Additionally, the mutation data identified 2 other potential contributing factors to MF, ultraviolet light, and a polymorphism in the tumor suppressor p53 (TP53). Therefore, genetic alterations in specific pathways in MF were identified that may be viable, effective new targets for treatment.


Amphibian thalamic nuclear organization during larval development and in the adult frog Xenopus laevis: Genoarchitecture and hodological analysis.
The early patterning of the thalamus during embryonic development defines rostral and caudal progenitor domains, which are conserved from fishes to mammals. However, the subsequent developmental mechanisms that lead to the adult thalamic configuration have only been investigated for mammals and other amniotes. In this study, we have analyzed in the anuran amphibian Xenopus laevis (an anamniote vertebrate), through larval and postmetamorphic development, the progressive regional expression of specific markers for the rostral (GABA, GAD67, Lhx1, and Nkx2.2) and caudal (Gbx2, VGlut2, Lhx2, Lhx9, and Sox2) domains. In addition, the regional distributions at different developmental stages of other markers such as calcium binding proteins and neuropeptides, helped the identification of thalamic nuclei. It was observed that the two embryonic domains were progressively specified and compartmentalized during premetamorphosis, and cell subpopulations characterized by particular gene expression combinations were located in periventricular, intermediate and superficial strata. During prometamorphosis, three dorsoventral tiers formed from the caudal domain and most pronuclei were defined, which were modified into the definitive nuclear configuration through the metamorphic climax. Mixed cell populations originated from the rostral and caudal domains constitute most of the final nuclei and allowed us to propose additional subdivisions in the adult thalamus, whose main afferent and efferent connections were assessed by tracing techniques under in vitro conditions. This study corroborates shared features of early gene expression patterns in the thalamus between Xenopus and mouse, however, the dynamic changes in gene expression observed at later stages in the amphibian support mechanisms different from those of mammals.


Development and validation of a product acceptability questionnaire for intranasal Q-Griffithsin COVID-19 prophylaxis (SPRAY PAL).
Patient experiences are critical when determining the acceptability of novel interventional pharmaceuticals. Here, we report the development and validation of a product acceptability questionnaire (SPRAY PAL) assessing feasibility, acceptability and tolerability of an intranasal Q-Griffithsin (Q-GRFT) drug product designed for COVID-19 prophylaxis. SPRAY PAL validation was undertaken as part of an ongoing phase 1 clinical trial designed to test the safety, pharmacokinetics and tolerability of intranasally administered Q-GRFT for the prevention of SARS-CoV-2 infection. The phase 1 clinical trial took place at a University Outpatient Clinical Trials Unit from November 2021 to September 2023. The initial SPRAY PAL questionnaire was piloted among healthy volunteers ages 25 to 55 in phase 1a of the clinical trial (N=18) and revised for administration in phase 1b for participants ages 24-59 (N=22). Spearman correlations tested convergent and discriminant validity. Internal consistency was assessed using Cronbach's alpha, and test-retest reliability was assessed using intraclass correlation coefficients of responses collected from three repeated questionnaire administrations. The initial version demonstrated excellent internal consistency. The revised version demonstrated very good internal consistency after removal of one item (alpha=0.739). Excellent test-retest reliability (intraclass coefficient=0.927) and adequate convergent (r's=0.208-0.774) and discriminant (r's=0.123-0.392) validity were achieved. Subscales adequately distinguished between the constructs of acceptability, feasibility and tolerability. The SPRAY PAL product acceptability questionnaire is a valid and reliable patient-reported outcomes measure that can be considered a credible tool for assessing patient-reported information about product acceptability, feasibility of use, tolerability of product and side effects and cost of product for novel intranasal drug formulations. The SPRAY PAL is generalisable, and items may be readily adapted to assess other intranasal formulations. NCT05122260 and NCT05437029.


The genetics of cutaneous melanoma.
Although germline mutations in CDKN2A are present in approximately 25% of large multicase melanoma families, germline mutations are much rarer in the smaller melanoma families that make up most individuals reporting a family history of this disease. In addition, only three families worldwide have been reported with germline mutations in a gene other than CDKN2A (i.e., CDK4). Accordingly, current genomewide scans underway at the National Human Genome Research Institute hope to reveal linkage to one or more chromosomal regions, and ultimately lead to the identification of novel genes involved in melanoma predisposition. Both CDKN2A and PTEN have been identified as genes involved in sporadic melanoma development; however, mutations are more common in cell lines than uncultured tumors. A combination of cytogenetic, molecular, and functional studies suggests that additional genes involved in melanoma development are located to chromosomal regions 1p, 6q, 7p, 11q, and possibly also 9p and 10q. With the near completion of the human genome sequencing effort, combined with the advent of high throughput mutation analyses and new techniques including cDNA and tissue microarrays, the identification and characterization of additional genes involved in melanoma pathogenesis seem likely in the near future.


Auditory brainstem response in a harbor porpoise show lack of automatic gain control for simulated echoes.
The auditory brainstem response (ABR) response to simulated echolocation clicks was studied in a harbor porpoise, Phocoena phocoena, to determine the relationship between the animal's perceived echo strength and the simulated target distance. In one experiment the click level at the listening post was kept constant while delay was changed, in another, the level was varied to approximate spreading losses. Results of both experiments indicate that there is no automatic gain control in the hearing system of this harbor porpoise.


The Functional Role of Prostate Cancer Metastasis-related Micro-RNAs.
The mortality of patients with hormone-resistant prostate cancer can be ascribed to a large degree to metastasis to distant organs, predominantly to the bones. In this review, we discuss the contribution of micro-RNAs (miRs) to the metastatic process of prostate cancer. The criteria for selection of miRs for this review were the availability of preclinical in vivo metastasis-related data in conjunction with prognostic clinical data. Depending on their function in the metastatic process, the corresponding miRs are up- or down-regulated in prostate cancer tissues when compared to matching normal tissues. Up-regulated miRs preferentially target suppressors of cytokine signaling or tumor suppressor-related genes and metastasis-inhibitory transcription factors. Down-regulated miRs promote epithelial-mesenchymal transition or mesenchymal-epithelial transition and diverse pro-metastatic signaling pathways. Some of the discussed miRs exert their function by simultaneously targeting epigenetic pathways as well as cell-cycle-related, anti-apoptotic and signaling-promoting targets. Finally, we discuss potential therapeutic options for the treatment of prostate cancer-related metastases by substitution or inhibition of miRs.


A curve evolution approach to object-based tomographic reconstruction.
In this paper, we develop a new approach to tomographic reconstruction problems based on geometric curve evolution techniques. We use a small set of texture coefficients to represent the object and background inhomogeneities and a contour to represent the boundary of multiple connected or unconnected objects. Instead of reconstructing pixel values on a fixed rectangular grid, we then find a reconstruction by jointly estimating these unknown contours and texture coefficients of the object and background. By designing a new "tomographic flow", the resulting problem is recast into a curve evolution problem and an efficient algorithm based on level set techniques is developed. The performance of the curve evolution method is demonstrated using examples with noisy limited-view Radon transformed data and noisy ground-penetrating radar data. The reconstruction results and computational cost are compared with those of conventional, pixel-based regularization methods. The results indicate that the curve evolution methods achieve improved shape reconstruction and have potential computation and memory advantages over conventional regularized inversion methods.


Adaptive independent vector analysis for multi-subject complex-valued fMRI data.
Complex-valued fMRI data can provide additional insights beyond magnitude-only data. However, independent vector analysis (IVA), which has exhibited great potential for group analysis of magnitude-only fMRI data, has rarely been applied to complex-valued fMRI data. The main challenges in this application include the extremely noisy nature and large variability of the source component vector (SCV) distribution. To address these challenges, we propose an adaptive fixed-point IVA algorithm for analyzing multiple-subject complex-valued fMRI data. We exploited a multivariate generalized Gaussian distribution (MGGD)- based nonlinear function to match varying SCV distributions in which the MGGD shape parameter was estimated using maximum likelihood estimation. To achieve our de-noising goal, we updated the MGGD-based nonlinearity in the dominant SCV subspace, and employed a post-IVA de-noising strategy based on phase information in the IVA estimates. We also incorporated the pseudo-covariance matrix of fMRI data into the algorithm to emphasize the noncircularity of complex-valued fMRI sources. Results from simulated and experimental fMRI data demonstrated the efficacy of our method. Our approach exhibited significant improvements over typical complex-valued IVA algorithms, especially during higher noise levels and larger spatial and temporal changes. As expected, the proposed complex-valued IVA algorithm detected more contiguous and reasonable activations than the magnitude-only method for task-related (393%) and default mode (301%) spatial maps. The proposed approach is suitable for decomposing multi-subject complex-valued fMRI data, and has great potential for capturing additional subject variability.


Sympathetic activation in the pathogenesis of hypertension and progression of organ damage.
Although animal models of hypertension have clearly shown that high blood pressure is associated with and is probably caused by an increase in sympathetic cardiovascular influences, a similar demonstration in humans has been more difficult to obtain for methodological reasons. There is now evidence, however, of increased sympathetic activity in essential hypertension. This article will review this evidence by examining data showing that plasma norepinephrine is increased in essential hypertension and that this is also the case for systemic and regional norepinephrine spillover, as well as for the sympathetic nerve firing rate in the skeletal muscle nerve district. Evidence will also be provided that sympathetic activation is a peculiar feature of essential hypertension, particularly in its early stages, with secondary forms of high blood pressure not usually characterized by an increased central sympathetic outflow. Humoral, metabolic, reflex, and central mechanisms are likely to be the factors responsible for the adrenergic activation characterizing hypertension, which may also promote the development and progression of the cardiac and vascular alterations that lead to hypertension-related morbidity and mortality, independent of blood pressure values. This represents the rationale for considering sympathetic deactivation one of the major goals of antihypertensive treatment.


Clustered lot quality assurance sampling: a pragmatic tool for timely assessment of vaccination coverage.
To evaluate oral poliovirus vaccine (OPV) coverage of the November 2009 round in five Northern Nigeria states with ongoing wild poliovirus transmission using clustered lot quality assurance sampling (CLQAS). We selected four local government areas in each pre-selected state and sampled six clusters of 10 children in each Local Government Area, defined as the lot area. We used three decision thresholds to classify OPV coverage: 75-90%, 55-70% and 35-50%. A full lot was completed, but we also assessed in retrospect the potential time-saving benefits of stopping sampling when a lot had been classified. We accepted two local government areas (LGAs) with vaccination coverage above 75%. Of the remaining 18 rejected LGAs, 11 also failed to reach 70% coverage, of which four also failed to reach 50%. The average time taken to complete a lot was 10 h. By stopping sampling when a decision was reached, we could have classified lots in 5.3, 7.7 and 7.3 h on average at the 90%, 70% and 50% coverage targets, respectively. Clustered lot quality assurance sampling was feasible and useful to estimate OPV coverage in Northern Nigeria. The multi-threshold approach provided useful information on the variation of IPD vaccination coverage. CLQAS is a very timely tool, allowing corrective actions to be directly taken in insufficiently covered areas.


Biomechanical characterization of myofibrillar myopathies.
Myofibrillar myopathies (MFMs) are a group of sporadic and hereditary skeletal muscle diseases, which lead to severe physical disability and premature death. Most MFMs are caused by mutations in genes encoding desmin, plectin, VCP, filamin C, BAG3, FHL-1, αB-crystallin, DNAJB6, myotilin, and ZASP. Biomechanical studies on primary human myoblasts carrying desmin and plectin mutations showed increased stiffness and reduced mechanical stress tolerance i.e., higher mechanical vulnerability compared to control cells. Higher stiffness of mutant cells may lead to higher intracellular stress at physiologic stretch and shear deformation, which in turn could trigger muscle fiber degeneration.


[Clinical and experimental studies on effects of chronic hepatitis B treated with astragali composita].
Eighty-seven cases of chronic persistent hepatitis (CPH) and 29 cases of chronic active hepatitis (CAH) were treated with Astragali Composita (AC), the clinical markedly effective rates were 69.0% and 55.2%, the total effective rates were 90.8% and 89.7% respectively; the sero-negative conversion rates of HBeAg and HBV-DNA were 27.7% and 28.0% respectively, they were higher than that of control group (P < 0.05-0.01). Eight ducks infected hepatitis B virus were treated with AC, sero-negative conversion of DHBV-DNA was seen in 3 ducks, DHBV-DNA levels of duck sera were lower after treatment than that before treatment (P < 0.05); mild inflammatory changes in liver tissues were seen in 3 ducks, its pathologic changes were milder than that of control groups; DHBV-DNA in liver tissues of 2 ducks was negative in situhybridization. The results indicated that AC had obvious effects of protecting liver tissues and some effects of inhibiting sera HBV and HBV replication.


Oxygen supersaturation protects coastal marine fauna from ocean warming.
Ocean warming affects the life history and fitness of marine organisms by, among others, increasing animal metabolism and reducing oxygen availability. In coastal habitats, animals live in close association with photosynthetic organisms whose oxygen supply supports metabolic demands and may compensate for acute warming. Using a unique high-frequency monitoring dataset, we show that oxygen supersaturation resulting from photosynthesis closely parallels sea temperature rise during diel cycles in Red Sea coastal habitats. We experimentally demonstrate that oxygen supersaturation extends the survival to more extreme temperatures of six species from four phyla. We clarify the mechanistic basis of the extended thermal tolerance by showing that hyperoxia fulfills the increased metabolic demand at high temperatures. By modeling 1 year of water temperatures and oxygen concentrations, we predict that oxygen supersaturation from photosynthetic activity invariably fuels peak animal metabolic demand, representing an underestimated factor of resistance and resilience to ocean warming in ectotherms.


Naturally Occurring Genetic Variants of Human Acetylcholinesterase and Butyrylcholinesterase and Their Potential Impact on the Risk of Toxicity from Cholinesterase Inhibitors.
Acetylcholinesterase (AChE) is the physiologically important target for organophosphorus toxicants (OP) including nerve agents and pesticides. Butyrylcholinesterase (BChE) in blood serves as a bioscavenger that protects AChE in nerve synapses from inhibition by OP. Mass spectrometry methods can detect exposure to OP by measuring adducts on the active site serine of plasma BChE. Genetic variants of human AChE and BChE do exist, but loss of function mutations have been identified only in the BCHE gene. The most common AChE variant, His353Asn (H322N), also known as the Yt blood group antigen, has normal AChE activity. The most common BChE variant, Ala567Thr (A539T) or the K-variant in honor of Werner Kalow, has 33% reduced plasma BChE activity. The genetic variant most frequently associated with prolonged response to muscle relaxants, Asp98Gly (D70G) or atypical BChE, has reduced activity and reduced enzyme concentration. Early studies in young, healthy males, performed at a time when it was legal to test nerve agents in humans, showed that individuals responded differently to the same low dose of sarin with toxic symptoms ranging in severity from minimal to moderate. Additionally, animal studies indicated that BChE protects from toxicants that have a higher reactivity with AChE than with BChE (e.g., nerve agents) but not from toxicants that have a higher reactivity with BChE than with AChE (e.g., OP pesticides). As a corollary, we hypothesize that individuals with genetic variants of BChE may be at increased risk of toxicity from nerve agents but not from OP pesticides.


Increased cellular free cholesterol in macrophage-specific Abca1 knock-out mice enhances pro-inflammatory response of macrophages.
Macrophage-specific Abca1 knock-out (Abca1(-)(M)(/-)(M)) mice were generated to determine the role of macrophage ABCA1 expression in plasma lipoprotein concentrations and the innate immune response of macrophages. Plasma lipid and lipoprotein concentrations in chow-fed Abca1(-)(M)(/-)(M) and wild-type (WT) mice were indistinguishable. Compared with WT macrophages, Abca1(-)(M)(/-)(M) macrophages had a >95% reduction in ABCA1 protein, failed to efflux lipid to apoA-I, and had a significant increase in free cholesterol (FC) and membrane lipid rafts without induction of endoplasmic reticulum stress. Lipopolysaccharide (LPS)-treated Abca1(-)(M)(/-)(M) macrophages exhibited enhanced expression of pro-inflammatory cytokines and increased activation of the NF-kappaB and MAPK pathways, which could be diminished by silencing MyD88 or by chemical inhibition of NF-kappaB or MAPK. In vivo LPS injection also resulted in a higher pro-inflammatory response in Abca1(-)(M)(/-)(M) mice compared with WT mice. Furthermore, cholesterol depletion of macrophages with methyl-beta-cyclodextrin normalized FC content between the two genotypes and their response to LPS; cholesterol repletion of macrophages resulted in increased cellular FC accumulation and enhanced cellular response to LPS. Our results suggest that macrophage ABCA1 expression may protect against atherosclerosis by facilitating the net removal of excess lipid from macrophages and dampening pro-inflammatory MyD88-dependent signaling pathways by reduction of cell membrane FC and lipid raft content.


Selection of cyclic peptide aptamers to HCV IRES RNA using mRNA display.
The hepatitis C virus (HCV) is a positive strand RNA flavivirus that is a major causative agent of serious liver disease, making new treatment modalities an urgent priority. Because HCV translation initiation occurs by a mechanism that is fundamentally distinct from that of host mRNAs, it is an attractive target for drug discovery. The translation of HCV mRNA is initiated from an internal ribosomal entry site (IRES), independent of cap and poly(A) recognition and bypassing eIF4F complex formation. We used mRNA display selection technology combined with a simple and robust cyclization procedure to screen a peptide library of >10(13) different sequences and isolate cyclic peptides that bind with high affinity and specificity to HCV IRES RNA. The best peptide binds the IRES with subnanomolar affinity, and a specificity of at least 100-fold relative to binding to several other RNAs of similar length. The peptide specifically inhibits HCV IRES-initiated translation in vitro with no detectable effect on normal cap-dependent translation initiation. An 8-aa cyclic peptide retains most of the activity of the full-length 27-aa bicyclic peptide. These peptides may be useful tools for the study of HCV translation and may have potential for further development as an anti-HCV drug.


Cardiobacterium valvarum infective endocarditis and phenotypic/molecular characterization of 11 Cardiobacterium species strains.
Cardiobacterium valvarum is a newly recognized human pathogen related to infective endocarditis. Cardiobacterium species are, however, only rarely the aetiology of infective endocarditis. An infective endocarditis case is presented and, additionally, phenotypic and phylogenetic comparison of a further 10 collection strains, representing the two species within the genus, was performed. C. valvarum was isolated from the blood and DNA was present in valvular tissue (partial 16S rRNA gene analysis) from a 64-year-old man with infective endocarditis of the mitral valve, rupture of chordae and prolapse of pulmonary valves in addition to a fluttering excrescence. A mechanical mitral valve and neochordae were inserted successfully. Phenotypically, the two species within the genus Cardiobacterium resemble each other greatly. When using the Vitek 2 Neisseria-Haemophilus identification card, the reaction for phenylphosphonate was positive for all Cardiobacterium hominis strains, but negative for all C. valvarum strains, thereby separating the two species. The two species made up two separate clusters by phylogenetic examination using 16S rRNA gene sequence analysis.


Chromatin fluorescence after carmine staining.
After staining with dilute solutions (0.1 mg/ml in distilled water) of commercial carmine, a strong reddish orange fluorescence was observed in nuclei from cell smears and frozen and paraffin tissue sections. Optimal exciting light was 436 nm (violet-blue) or 450-490 nm (blue). Compact chromatin from interphase nuclei, mitotic and meiotic chromosomes and the kinetoplast of Trypanosoma cruzi showed the highest fluorescence, while the basophilic cytoplasm appeared weakly fluorescent. No emission was observed in cartilage matrix, mast cell granules or goblet cell mucin. This selective method could be valuable in microscopic and cytochemical studies on chromatin because the carmine fluorescence is stable and preparations can be dehydrated and mounted permanently without changes in the fluorescence pattern.


Analysis of circulating microRNAs during adjuvant chemotherapy in patients with luminal A breast cancer.
Breast cancer is a complex disease displaying different profiles involving genetic as well as epigenetic factors. MicroRNAs (miRNAs) are small, non-coding RNAs that regulate gene expression. Recent studies demonstrated that miRNAs may display great potential for the development of novel biomarkers and therapeutic targets. In the present study, the levels of miR-21 and miR-145 were analyzed in the peripheral blood of 52 patients with luminal A breast cancer. miRNA expression was determined in serum samples from matched pre- and post-treatment patients with breast cancer by quantitative polymerase chain reaction. There were no statistically significant differences in miR-145 and miR-21 levels between pre- and post-treatment samples. In addition, the miRNA levels were not found to be associated with the clinical parameters.


Role of HMGB1 in proliferation and migration of human gingival and periodontal ligament fibroblasts.
High mobility group box 1 (HMGB1) was originally defined as a nuclear protein. However, later studies showed that HMGB1 was released from damaged cells into the extracellular milieu and functioned as a danger signaling molecule. HMGB1 has also been shown to exert proliferative and chemoattractant effects on many cell types. In this study, we investigated the in vitro effect of human recombinant HMGB1 on the proliferation and migration of human gingival fibroblasts (HGF) and human periodontal ligament fibroblasts (HPDLF). For the proliferation assay, HGF and HPDLF were cultured in the presence of 5, 10, and 50 ng/mL HMGB1. After a period of 6 days, cell proliferation was determined by MTT assay. The migration assay was performed by culturing the two cell types in Transwells with HMGB1 in the lower chamber as a chemoattractant. Cell migration during 16 h was determined by crystal violet staining of the cells that migrated across the membrane. The results showed that HMGB1, at 50 ng/mL, was able to significantly induce proliferation of HGF by up to 171.4 ± 17.1%. No such proliferation induction was seen for HPDLF. In the migration assay, however, 100 ng/mL HMGB1 induced migration of both cell types. The counts of cells that migrated across the membrane, as compared with the control, were increased to 273 ± 24.1% and 410.3 ± 158% for HGF and HPDLF, respectively. Since proliferation and migration are basic abilities of cells required for proper tissue repair, these data suggest that HMGB1 plays an important role in these functions of periodontal cells.


In defense of the salience map: salience rather than visibility determines selection.
The aim of the present study was to investigate whether time-dependent biases of oculomotor selection as typically observed during visual search are better accounted for by an absolute-processing-speed account (J. P. de Vries, I. T. C. Hooge, M. A. Wiering, & F. A. J. Verstraten, 2011, How longer saccade latencies lead to a competition for salience. Psychological Science, 22, 916-923) or a relative-salience account (e.g., M. Donk, & W. van Zoest, 2008, Effects of salience are short-lived. Psychological Science, 19, 733-739; M. Donk & W. van Zoest, 2011, No control in orientation search: The effects of instruction on oculomotor selection in visual search. Vision Research, 51, 2156-2166). In order to test these two models, we performed an experiment in which participants were instructed to make a speeded eye movement to any of two orientation singletons presented among a homogeneous set of vertically oriented background lines. One singleton, the fixed singleton, remained identical across conditions, whereas the other singleton, the variable singleton, varied such that its orientation contrast relative to the background lines was either smaller or larger than that of the fixed singleton. The results showed that the proportion of eye movements directed toward the fixed singleton varied substantially depending on the orientation contrast of the variable singleton. A model assuming selection behavior to be determined by relative salience provided a better fit to the individual data than the absolute processing speed model. These findings suggest that relative salience rather than the visibility of an element is crucial in determining temporal variations in oculomotor selection behavior and that an explanation of visual selection behavior is insufficient without the concept of a salience map.


Urinary excretion of advanced glycation end products in dogs and cats.
The present study was conducted with privately owned dogs and cats to investigate whether a relationship exists between the dietary AGEs and the urinary excretion of AGEs, as indication of possible effective absorption of those compounds in the intestinal tract of pet carnivores. For this purpose, data were collected from both raw fed and dry processed food (DPF) fed to dogs and cats, through spot urine sampling and questionnaires. Raw pet food (RF, low in AGE diets) was fed as a primary food source to 29 dogs and DPF to 28 dogs. Cats were categorized into 3 groups, which were RF (n = 15), DPF (n = 14) and dry and wet processed pet food (DWF, n = 25). Urinary-free carboxymethyllysine (CML), carboxyethyllysine (CEL) and lysinoalanine (LAL) were analysed using ultrahigh-performance liquid chromatography (UHPLC)-mass spectrometry, and were standardized for variable urine concentration by expressing the AGE concentrations as a ratio to urine creatinine (Ucr) concentration (µg/µmol Ucr). Urinary excretion of CML, CEL and LAL in dogs fed with DPF was 2.03, 2.14 and 3 times higher compared to dogs fed with RF (p < .005). Similar to the dogs, a significant difference in CML:Ucr, CEL:Ucr and LAL:Ucr between the three diet groups was observed in cats (p-overall < 0.005, ANOVA), in which the RF fed group excreted less AGEs than the other groups. Linear regression coefficients and SE of CML:Ucr, CEL:Ucr and LAL:Ucr showed that body weight and neuter status were significantly correlated with CML and CEL excretion, but not to LAL excretion. Our results revealed a significant correlation between dietary AGEs and urinary excretion of free CML, CEL and LAL, and also showed that endogenous formation of these AGEs occurs in both dogs and cats under physiological conditions.


Steroidogenesis in adrenal tumor cells: influence of cell shape.
Y-1 adrenal tumor cells were grown on plastic, or plastic treated with poly(2-hydroxyethyl methacrylate) (polyHEMA) to produce concentration-dependent rounding (10(-5)-3 X 10(-4) M) of the cells or on plastic treated with poly-D-lysine (polylysine) to produce flat cells, in order to determine whether or not cell shape is correlated with steroid synthesis. The degree of rounding of cells was measured by determining mean cell height and longest cell diameter. Three measurements of steroid production were made: production of 20 alpha-dihydroprogesterone, transport of cholesterol to the inner mitochondrial membrane, and production of pregnenolone by isolated mitochondria. Cells grown on poly(HEMA) showed increase in mean cell height, decrease in longest diameter (i.e. rounding), and increase in all three measurements of steroidogenesis. In the case of synthesis of 20 alpha-dihydroprogesterone, the response was dependent on the concentration of poly(HEMA), being greater with higher concentrations (up to 10(-4) M), of this agent. Moreover the degree of rounding (cell height) was correlated with production of 20 alpha-dihydroprogesterone at three concentrations of poly(HEMA) (r = 0.93. ACTH at a supramaximal concentration produced increases in all of these responses to the poly(HEMA) surface. Polylysine produced flatter cells (lower mean height and greater longest diameter) than plastic and also inhibited all three steroidogenic responses to ACTH. (Bu)2cAMP exerted the same effects as ACTH. Growing cells on poly(HEMA) or polylysine did not affect production of cyclic AMP by the cells. Addition of poly(HEMA) or polylysine to the medium in which the cells were incubated, at the same concentrations as those used for influencing cell shape, was without effect on steroid synthesis or the response to ACTH. Cells grown on poly(HEMA) show decreased incorporation of [3H] thymidine into DNA. It is concluded that cell shape influences the delivery of cholesterol to inner mitochondrial membrane and in this way, increases the production of steroids by Y-1 cells and that the effects of poly(HEMA) on cell shape, cholesterol transport, and synthesis of DNA may involve microfilaments.


Which Characteristics are Associated with the Timing of the First Healthcare Consultation, and Does the Time to Care Influence the Duration of Compensation for Occupational Back Pain?
Purpose To identify the characteristics associated with the timing of the first healthcare consultation and to measure the impact of that timing on the duration of the first episode of compensation for occupational back pain following the injury. Methods We analyzed data from a cohort of workers with compensated back pain in 2005 in Ontario obtained from the Workplace Safety and Insurance Board. Cox multivariable survival models were performed to identify factors associated with the time to care and to measure its association with the length of the first episode of 100 % compensation. Results Among the 5520 claims analyzed, 93.7 % of workers accessed care within the first week (average = 2.1 days; median = 1 day). Time to care was shorter for males, for workers who had received previous compensation and for those with access to an early return to work program. Age, number of employees in the company and personal earnings were positively associated with the time to care. More severe nature of injury, employers doubt about the work-relatedness of the injury and consulting a physiotherapist as the first healthcare provider were also associated with longer time to care. Considering potential confounders, longer time to care was significantly associated with a delay in the end of the first episode of compensation (hazard ratio = 0.98; P < 0.001). Conclusion Temporal access to a source of care is not problematic for the vast majority of Ontarian workers who receive compensation for occupational back pain; however, for the minority of workers who do not rapidly access care, the timing of the first healthcare consultation is a significant predictor of the duration of the first episode of compensation.


Clinical Characteristics and Risk Factors of Cerebral Hemorrhage in Patients with Occult Malignant Tumors.
To investigate the clinical characteristics and potential risk factors of the first onset of cerebral hemorrhage in patients with occult malignant tumors. In this retrospective study, 23 patients with occult malignant tumors with the first onset of cerebral hemorrhage were enrolled in the tumor group, and 92 patients without occult tumors in the same period were enrolled in the control group. There were no statistical differences in age and sex between both groups by propensity score matching. Collected clinical data included age, sex, smoking history, drinking history, hypertension history, diabetes history, past medical history, routine blood tests, neutrophil-to-lymphocyte ratio (NLR), liver and kidney function, fasting blood glucose level, coagulation function, tumor markers, imaging examinations, National Institute of Health stroke scale (NIHSS) score on admission, modified Rankin Scale (mRS) score 90 days after intracerebral hemorrhage and final mRS score. Compared with the control group, the tumor group had fewer patients with hypertension (52.2% vs 81.5%, P<0.05), and the NLR was significantly decreased in the tumor group (2.74 vs 5.46, P<0.05). The tumor group had a greater number of patients with the bleeding sites located in the lobar regions (43.5% vs.19.6%, P<0.05) and a higher coagulation dysfunction (52.2% vs 29.3%, P<0.05) than the control group. Multivariate logistic regression analysis revealed that no history of hypertension (OR: 3.141, 95% CI: 1.107-8.916), lobar cerebral hemorrhage (OR: 3.465 95% CI:1.172-10.243), and coagulation dysfunction (OR: 3.176, 95% CI: 1.131-8.913) were independent predictors of occult tumors, and the receiver operating characteristic (ROC) curve showed that the area under the curve of the three-index combined diagnosis was 0.748, C-statistic analysis also showed the same result. No history of hypertension, lobar cerebral hemorrhage, and coagulation dysfunction may be predictors of the risk of occult malignancies in patients with cerebral hemorrhage.


Induction of cells with phenotypic features of neuronal cells by treatment with dibutyryl cyclic adenosine 3',5'-monophosphate in a human parotid gland adenocarcinoma cell line in culture.
A human parotid gland adenocarcinoma cell line, with an intercalated duct cell phenotype of the salivary gland and expression of vasoactive intestinal polypeptide and amylase, was cultivated in the presence of dibutyryl cyclic adenosine 3',5'-monophosphate (dB-cAMP). Morphological changes occurred; cells formed long cytoplasmic processes densely packed with ample microfibrils, as well as microtubules, and grew in a netlike appearance. In addition, it has been found by the immunofluorescence staining technique, immunoblotting, or immunoelectron microscopy that the cells treated with dB-cAMP express neurofilaments, neuron-specific enolase, synaptophysin, and HNK-1 antigen, as well as the alpha- and beta-chains of tubulin, whereas these antigens are not detected in untreated cells. The expression of vasoactive intestinal polypeptide detected diffusely in the cytoplasm of untreated cells was restricted to the cell membranes during the cultivation of cells in the presence of dB-cAMP, while expression of amylase persisted in the treated cells in a fashion similar to that in untreated cells. Moreover, both anchorage-independent and anchorage-dependent growth of the cells was markedly suppressed in the presence of dB-cAMP. After removal of dB-cAMP from the culture, the treated cells returned rapidly to the phenotype and growth rate of the untreated cells. These findings indicate that reversible conversion into cells with phenotypic features of neuronal cells of a human parotid adenocarcinoma cell line occurs in growth medium containing dB-cAMP.


Contrasting Patterns of Virus Protection and Functional Incompatibility Genes in Two Conspecific Wolbachia Strains from Drosophila pandora.
Wolbachia infections can present different phenotypes in hosts, including different forms of reproductive manipulation and antiviral protection, which may influence infection dynamics within host populations. In populations of Drosophila pandora two distinct Wolbachia strains coexist, each manipulating host reproduction: strain wPanCI causes cytoplasmic incompatibility (CI), whereas strain wPanMK causes male killing (MK). CI occurs when a Wolbachia-infected male mates with a female not infected with a compatible type of Wolbachia, leading to nonviable offspring. wPanMK can rescue wPanCI-induced CI but is unable to induce CI. The antiviral protection phenotypes provided by the wPanCI and wPanMK infections were characterized; the strains showed differential protection phenotypes, whereby cricket paralysis virus (CrPV)-induced mortality was delayed in flies infected with wPanMK but enhanced in flies infected with wPanCI compared to their respective Wolbachia-cured counterparts. Homologs of the cifA and cifB genes involved in CI identified in wPanMK and wPanCI showed a high degree of conservation; however, the CifB protein in wPanMK is truncated and is likely nonfunctional. The presence of a likely functional CifA in wPanMK and wPanMK's ability to rescue wPanCI-induced CI are consistent with the recent confirmation of CifA's involvement in CI rescue, and the absence of a functional CifB protein further supports its involvement as a CI modification factor. Taken together, these findings indicate that wPanCI and wPanMK have different relationships with their hosts in terms of their protective and CI phenotypes. It is therefore likely that different factors influence the prevalence and dynamics of these coinfections in natural Drosophila pandora hosts.IMPORTANCEWolbachia strains are common endosymbionts in insects, with multiple strains often coexisting in the same species. The coexistence of multiple strains is poorly understood but may rely on Wolbachia organisms having diverse phenotypic effects on their hosts. As Wolbachia is increasingly being developed as a tool to control disease transmission and suppress pest populations, it is important to understand the ways in which multiple Wolbachia strains persist in natural populations and how these might then be manipulated. We have therefore investigated viral protection and the molecular basis of cytoplasmic incompatibility in two coexisting Wolbachia strains with contrasting effects on host reproduction.