Caloric restriction upregulates the expression of DNMT3.1, lacking the conserved catalytic domain, in Daphnia magna.
DNA methylation plays an important role in many aspects of biology, including development, disease, and phenotypic plasticity. In the branchiopod crustacean, Daphnia, de novo DNA methylation has been detected in specific environmental contexts. However, fundamental information on de novo DNA methyltransferase DNMT3 orthologs, including domain organization, developmental expression, and response to environmental stimuli, is lacking. In this study, we examined two DNMT3 orthologs in Daphnia magna, DapmaDNMT3.1 and DapmaDNMT3.2. Amino acid sequence alignment revealed that DapmaDNMT3.1 and DapmaDNMT3.2 lack the conserved methyltransferase motifs of the catalytic domain and the PWWP domain, respectively. We profiled the expression of the two orthologs during embryogenesis and under various feeding levels. During embryogenesis, in contrast to the low DapmaDNMT3.1 expression, DapmaDNTM3.2 was highly expressed at specific stages, that is, in the one cell-stage and at 48 hr post ovulation. In nutrient-rich condition, both genes were lowly expressed, whereas DapmaDNMT3.1 was upregulated at the lower food levels, suggesting a potential role of DapmaDNMT3.1 in gene regulation in response to caloric restriction. These findings provide a basis for understanding the developmental stage- and stress-dependent function of DNMT3 orthologs in D. magna.


The longitudinal association between lifetime mental disorders and first onset or recurrent suicide ideation.
Although the cross-sectional association between mental disorders and suicide ideation is well studied, less is known about the prospective association. In this paper, we estimated among those without 12-month suicide ideation at baseline, the association between a wide variety of common mental disorders at baseline and suicide ideation within the 6-year follow-up period, after controlling for history of other mental disorders and demographic variables. Data were used from the Netherlands Mental Health Survey and Incidence Study-2 (NEMESIS-2), a prospective representative adult cohort study with baseline (n = 6646) with a 6-year follow-up period. Lifetime mental disorders were assessed at baseline with the Composite International Diagnostic Interview 3.0. Within the longitudinal design, participants with first time or recurrent suicide ideation were defined follows: having no suicide ideation in the 12 months before the baseline assessment, and reporting to have had seriously thought about suicide between baseline and the 6-year follow-up period. Multiple logistical regression was used to estimate the longitudinal association between suicide ideation and a specific mental disorder while controlling for comorbidity and baseline variables. To account for the prevalence of a disorder in the population, for each disorder, the population attributable risk proportion (PARP) was calculated. 2.9% (n = 132) of the participants that did not report suicide ideation in the past 12 months at baseline reported suicide ideation at follow-up. Of these 132 cases, 81 (61%) experienced suicide ideation for the first time in their lives and could be viewed as first onset cases. 51 (39%) reported recurrent suicide ideation. After controlling for comorbidity, the only two disorders that were significantly related to suicide ideation at follow-up were lifetime major depressive disorder (MDD) and generalized anxiety disorder (GAD). PARP for MDD was 47.8 and 16.6% for GAD. After controlling for all other mental disorders, a lifetime history of MDD and GAD were related to suicide ideation at follow-up. For clinical practice, this indicates that patients with a history of MDD or GAD stay vulnerable for suicide ideation, even though they did not report suicide ideation in the past year.


[Blood vasopressin concentration is patients of different ages with hypertension].
The concentration of blood vasopressin was investigated in apparently healthy persons and in patients with I--II degree hypertension, aged from 20 to 80 years. Vasopressin concentration was determined by the biological method according to the antidiuretic effect of ethanol-anesthetized and constantly hydrated rats on an original 5-channel apparatus. The results obtained showed the blood vasopressin concentration to increase with age. In patients with the I--II degree hypertensive disease the mentioned concentration was significantly higher than in healthy persons of the same age. Close correlation coefficient was revealed between the blood vasopressin concentration and minimal arterial blood pressure values.


Autonomic neuropathy in streptozotocin diabetic rats: effect of acetyl-L-carnitine.
The present study was designed to characterize cardiac autonomic neuropathy in streptozotocin-induced (45 mg/kg i.v.) diabetic rat by analysis of heart rate variability (HRV), and to assess, in this model, the effects of treatment with acetyl-L-carnitine (ALC). Heart rate was reduced in diabetic rats (332+/-22 vs. 411+/-35 beat per min; P<0.0001). This bradycardia was partly reversed with ALC (369+/-52 beat per min; P<0.05 vs. untreated). Both time- and frequency-domain parameters of HRV were significantly reduced in diabetic rats. The reduction of spectral power was around 50% at high frequencies and about 70% at low frequencies, suggesting a decrease of parasympathetic activity. Low/high frequency ratio was significantly decreased in diabetic rats suggesting decreased sympathetic tone, while nonlinear analysis indicated a reduction of the chaotic complexity of heart rate dynamics in diabetic rats. Standard deviation of heart rate in ALC-treated rats was significantly higher than in untreated diabetic rats (P<0.0001). ALC counteracts the reduction of the power spectrum observed in diabetic animals (P<0.0005) normalizing the spectra profile. ALC restored chaotic complexity of heart rate dynamics. These results on the whole indicate that both sympathetic and parasympathetic cardiac tone were reduced significantly in diabetic rats and that ALC treatment prevents the development of autonomic neuropathy in streptozotocin-induced diabetes in rats.


The origins and evolution of vertebrate metamorphosis.
Metamorphosis, classically defined as a spectacular post-embryonic transition, is well exemplified by the transformation of a tadpole into a frog. It implies the appearance of new body parts (such as the limbs), the resorption of larval features (such as the tail) and the remodelling of many organs (such as the skin or the intestine). In vertebrates, metamorphosis has been well characterized in anuran amphibians, where thyroid hormones orchestrate the intricate and seemingly contradictory changes observed at the cellular and tissue levels. Thyroid hormones control a complex hierarchical cascade of target genes via binding to specific receptors, TRα and TRβ, ligand-activated transcription factors belonging to the nuclear receptor superfamily. Metamorphosis is actually widespread in the vertebrates, though quite diverse in the way it manifests in a particular species. Furthermore, evolutionary and ecological variations of this key event, from paedomorphosis to direct development, provide an excellent illustration of how tinkering with a control pathway can lead to divergent life histories. The study of invertebrate chordates has also shed light on the origin of metamorphosis. The available data suggest that post-embryonic remodelling governed by thyroid hormones is an ancestral feature of chordates. According to this view, metamorphosis of the anurans is an extreme example of a widespread life history transition.


The transcription factor early growth response 1 (Egr-1) advances differentiation of pre-B and immature B cells.
In mature B lymphocytes, the zinc finger transcription factor early growth response 1 (Egr-1) is one of the many immediate-early genes induced upon B cell antigen receptor engagement. However, its role during earlier stages of lymphopoiesis has remained unclear. By examining bone marrow B cell subsets, we found Egr-1 transcripts in pro/pre-B and immature B lymphocytes, and Egr-1 protein in pro/pre-B-I cells cultivated on stroma cells in the presence of interleukin (IL)-7. In recombinase-activating gene (RAG)-2-deficient mice overexpressing an Egr-1 transgene in the B lymphocyte lineage, pro/pre-B-I cells could differentiate past a developmental block at the B220(low) BP-1(-) stage to the stage of B220(low) BP-1(+) pre-B-I cells, but not further to the B220(low) BP-1(+) CD25(+) stage of pre-B-II cells. Therefore, during early B lymphopoiesis progression from the B220(low) BP-1(-) IL-2R- pro/pre-B-I stage to the B220(low) BP-1(+) IL-2R+ pre-B-II stage seems to occur in at least two distinct steps, and the first step to the stage of B220(low) BP-1(+) pre-B-I cells can be promoted by the overexpression of Egr-1 alone. Wild-type mice expressing an Egr-1 transgene had increased proportions of mature immunoglobulin (Ig)M+ B220(high) and decreased proportions of immature IgM+ B220(low) bone marrow B cells. Since transgenic and control precursor B cells show comparable proliferation patterns, overexpression of Egr-1 seems also to promote entry into the mature B cell stage. Analysis of changes in the expression pattern of potential Egr-1 target genes revealed that Egr-1 enhances the expression of the aminopeptidase BP-1/6C3 in pre-B and immature B cells and upregulates expression of the orphan nuclear receptor nur77 in IgM+ B cells.


Role of macrophages in mesothelial healing.
The role of macrophages in rat mesothelial healing was evaluated in vivo and in vitro. In animals whose circulating monocytes were severely depressed, the healing rate was retarded; however, this effect was partly reversed by the introduction of macrophages into the injured site, suggesting that macrophages are of fundamental importance in the healing of mesothelial wounds. Furthermore, it was found that wound exudates stimulated DNA synthesis in normal quiescent mesothelial cells and the component principally responsible for this activity was the macrophage. Cultured macrophage-conditioned media also induced an increase in mesothelial replication. Moreover, the mitogenic activity present in both wound exudates and macrophage-conditioned media was increased by dialysis and diminished by heating at 80 degrees C for 1 h. The putative mesothelial mitogenic factor in the supernatant of wound exudates and macrophage cell cultures has a molecular size greater than 7000 dalton and is stable after mild heating (60 degrees C, 1 h). It is postulated that exudate macrophages secrete mitogenic factor(s) which stimulate mesothelial proliferation, and initiate healing.


[Primary treatment of acute extensor tendon injuries of the hand].
Reconstruction of extensor functions after extensor tendon injuries of the hand. Acute injuries of extensor mechanism with corresponding loss of function. Complex injuries with loss of soft tissue. Limited possibility of extensor tendon reconstruction with combined injuries of the interphalangeal joints (in situations with irreparable joints: primary arthrodesis). The treatment of extensor tendon injuries depends on the various levels of tendon laceration. Zones 1 and 2: in case of tendon disruption close to the base of the distal phalanx, refixation of tractus terminalis using a pull-out suture. In case of disruption more proximally, primary repair using mattress sutures. Temporary pinning of the distal interphalangeal joint in extension using a single transarticular Kirschner wire. Zone 3: mattress sutures of the tractus intermedius. Temporary pinning of the proximal interphalangeal joint in extension using a single transarticular Kirschner wire. Zone 4: reconstruction of the central slip and the lateral slip of extensor tendon using modified Becker sutures and mattress sutures. Temporary pinning of the proximal interphalangeal joint in extension using a single transarticular Kirschner wire. Zones 5 and 6: four-strand modified Becker sutures with additional epitendinous suture. Zones 7 and 8: core sutures using modified Kirchmayr techniques with additional epitendinous suture. Zones 1-4: immobilization of the finger for 6 weeks with removal of the transarticular wire at 4 weeks. Zones 5-8: dynamic postoperative treatment in intrinsic-plus splint for 6 weeks. It is postulated that dynamic postoperative treatment leads to improved functional outcome after extensor tendon injuries. While for zones 1-4 no better final clinical results are observed using the dynamic postoperative protocol, early protected motion for zones 5-8 is superior to static post operative treatment.


R-wave amplitude changes during exercise stress testing: patients with dilative cardiomyopathy compared to patients with coronary artery disease.
R-wave amplitude (RWA) depends to a large extent on the left ventricular filling volume. Changes of RWA are attributed to the Brody-effect. Exercise has been shown to induce a decrease of RWA in a healthy population and an increase in patients with coronary artery disease (CAD). No clear data exist for cardiomyopathy (CMP). Controls (n = 12), patients with CMP (n = 32) and CAD (n = 58) were compared. Alterations of RWA (Wilson lead V5) were correlated with parameters of a bicycle exercise test including resting and exercise hemodynamics and parameters of LV-function including EF, LVEDV and LVEDP. CMP compared to CAD had smaller RWA at rest (0.78 +/- 0.47 vs 1.32 +/- 0.72 mV, P less than 0.01). During comparable levels of exercise CMP (EF 35 +/- 14%) showed no significant changes of RWA. CAD (EF 57 +/- 16%) presented an increase of RWA by +0.11 +/- 0.23 mV (P less than 0.01), while controls showed a straight decline of RWA (-0.31 +/- 0.24 mV). In patients with CAD delta RWA (RWA max exercise - RWA rest) was a more sensitive parameter for detection of disease (assuming delta RWA greater than or equal to +0.1 mV, 36/58 patients) than maximal ST-segment changes (ST-segment-depression in lead V5 greater than or equal to 0.2 mV at 0.08 sec after J-point, 22/58 patients). Precordial leads V2, V4, V5 and V6 showed similar changes. This paper supports the theory that changes of wall thickness or changes in the amount of air respective to the amount of fluid in the lungs are responsible for RWA changes. These changes are clearly dependent on the severity of the disease and on left ventricular function. Therefore measurement of RWA changes during exercise may offer additional information in patients with CAD as well as in patients with CMP.


Influence of dose frequency on the therapeutic efficacies of ciprofloxacin and ceftazidime in experimental Klebsiella pneumoniae pneumonia and septicemia in relation to their bactericidal activities in vitro.
The antibacterial activities of ciprofloxacin versus ceftazidime against Klebsiella pneumoniae in vitro and in vivo were compared. Although there was only a minor difference in MBC values between both drugs ciprofloxacin demonstrated a high and dose-dependent bacterial killing rate in vitro and in lungs of leukopenic rats in contrast to the more time-dependent bactericidal activity of ceftazidime. After treatment of a K.pneumoniae pneumonia and septicemia the efficacy of ciprofloxacin was only slightly influenced by the mode of administration, either at 6-h intervals or continuously, whereas ceftazidime was far more effective after continuous administration. This resulted in a superior efficacy of ciprofloxacin after intermittent treatment as compared to ceftazidime, whereas ceftazidime was more effective after continuous administration as compared to ciprofloxacin. Also ciprofloxacin proved to be bactericidal against bacteria that were not actively growing, both in vitro and in vivo, whereas ceftazidime was not.


Event-related potentials during naming and mental rotation.
The biphasic late negative wave (Nx-Ny) evoked during the naming of pictures of objects was investigated in a series of experiments. The monitoring of several possible non-cerebral generators indicated that these components were probably cerebral in origin. The Ny wave differed in scalp distribution from the preceding CNV. It was unaffected by stimulus duration or by stimulus repetition. The Nx-Ny complex occurred in both a naming task and a mental rotation task. The slow waves following the biphasic negative wave differed significantly between these two tasks. In the mental rotation task, there was a large prolonged negativity in both parietal regions. The Nx wave (at approximately 250 msec) of the biphasic complex may represent the initial registration of the stimulus. The Ny wave (at approximately 420 msec) does not appear to be specifically related to semantic processing. Rather it may index some process involved in initiating the further evaluation of a complex stimulus. The slow waves that follow the biphasic negative complex may reflect the actual perceptual processing of the stimulus.


[Genetics of male infertility].
Genetic causes of male infertility increase in frequency with decreasing sperm concentration (oligo-/azoospermia). The decision about genetic tests should be made after a complete andrological work-up. Common causes comprise chromosomal aberrations (including Klinefelter syndrome), microdeletions of the AZF loci of the Y chromosome, mutations in the gene responsible for cystic fibrosis (CFTR) causing CBAVD and in genes involved in hypogonadotropic hypogonadism (including Kallmann syndrome). Every genetic investigation should be accompanied by comprehensive genetic counselling to help with the interpretation of results and support the patient/the couple concerning consequences for their family planning and treatment options.


The modulation effects of repeated transcutaneous auricular vagus nerve stimulation on the functional connectivity of key brainstem regions along the vagus nerve pathway in migraine patients.
Previous studies have shown a significant response to acute transcutaneous vagus nerve stimulation (taVNS) in regions of the vagus nerve pathway, including the nucleus tractus solitarius (NTS), raphe nucleus (RN) and locus coeruleus (LC) in both healthy human participants and migraine patients. This study aims to investigate the modulation effect of repeated taVNS on these brainstem regions by applying seed-based resting-state functional connectivity (rsFC) analysis. 70 patients with migraine were recruited and randomized to receive real or sham taVNS treatments for 4 weeks. fMRI data were collected from each participant before and after 4 weeks of treatment. The rsFC analyses were performed using NTS, RN and LC as the seeds. 59 patients (real group: n = 33; sham group: n = 29) completed two fMRI scan sessions. Compared to sham taVNS, real taVNS was associated with a significant reduction in the number of migraine attack days (p = 0.024) and headache pain intensity (p = 0.008). The rsFC analysis showed repeated taVNS modulated the functional connectivity between the brain stem regions of the vagus nerve pathway and brain regions associated with the limbic system (bilateral hippocampus), pain processing and modulation (bilateral postcentral gyrus, thalamus, and mPFC), and basal ganglia (putamen/caudate). In addition, the rsFC change between the RN and putamen was significantly associated with the reduction in the number of migraine days. Our findings suggest that taVNS can significantly modulate the vagus nerve central pathway, which may contribute to the potential treatment effects of taVNS for migraine.Clinical Trial Registration: http://www.chictr.org.cn/hvshowproject.aspx?id=11101, identifier ChiCTR-INR-17010559.


Biomarker variance component estimation for exposure surrogate selection and toxicokinetic inference.
Biomarkers are useful exposure surrogates given their ability to integrate exposures through all routes and to reflect interindividual differences in toxicokinetic processes. Also, biomarker concentrations tend to vary less than corresponding environmental measurements, making them less-biasing surrogates for exposure. In this article, urinary PAH biomarkers (namely, urinary naphthalene [U-Nap]; urinary phenanthrene [U-Phe]; 1-hydroxypyrene [1-OH-Pyr]; and 1-, (2+3)-, 4-, and 9-hydroxyphenanthrene [1-, (2+3)-, 4-, and 9-OH-Phe]) were evaluated as surrogates for exposure to hot asphalt emissions using data from 20 road-paving workers. Linear mixed-effects models were used to estimate the within- and between-person components of variance for each urinary biomarker. The ratio of within- to between-person variance was then used to estimate the biasing effects of each biomarker on a theoretical exposure-response relationship. Mixed models were also used to estimate the amounts of variation in Phe metabolism to individual OH-Phe isomers that could be attributed to Phe exposure (as represented by U-Phe concentrations) and covariates representing time, hydration level, smoking status, age, and body mass index. Results showed that 1-OH-Phe, (2+3)-OH-Phe, and 1-OH-Pyr were the least-biasing surrogates for exposure to hot asphalt emissions, and that effects of hydration level and sample collection time substantially inflated bias estimates for the urinary biomarkers. Mixed-model results for the individual OH-Phe isomers showed that between 63% and 82% of the observed biomarker variance was collectively explained by Phe exposure, the time and day of sample collection, and the hydration level, smoking status, body mass index, and age of each worker. By difference, the model results also showed that, depending on the OH-Phe isomer, a maximum of 6-23% of the total biomarker variance was attributable to differences in unobserved toxicokinetic processes between the workers. Therefore, toxicokinetic processes are probably less influential on urinary biomarker variance than are exposures and observable covariate effects. The methods described in this analysis should be considered for the selection and interpretation of biomarkers as exposure surrogates in future exposure investigations.


Auranofin, an oral chrysotherapeutic agent, inhibits histamine release from human basophils.
The effect of auranofin, an oral chrysotherapeutic agent, on histamine release from human basophils was studied. Auranofin inhibited IgE-mediated, anti-IgE-induced histamine release in a dose-dependent fashion with a concentration of drug required to produce 50% inhibition of 1.3 micrograms/ml. This compound also inhibited calcium ionophore A23187 and 12-0-tetradecanoyl-phorbol-13-acetate-induced histamine release with a concentration of drug required to produce 50% inhibition of 1.7 micrograms/ml and 0.9 micrograms/ml, respectively. Auranofin was less active for formyl-L-methionyl-L-leucyl-L-phenylalanine-induced release of histamine with 32 +/- 10% (mean +/- SEM) inhibition at 2 micrograms/ml. Auranofin effects were reversible when leukocytes preincubated with auranofin was washed with buffer. In contrast, gold sodium thiomalate enhanced IgE-mediated histamine release, suggesting that these two compounds exert their actions at different levels. These results suggest that auranofin may be beneficial to patients with allergic diseases such as bronchial asthma.


MRI of fast-relaxing spins.
MR imaging of extremely fast-relaxing spins is currently a topic of much interest due to recent technical innovations and groundbreaking studies demonstrating its utility in biomedical research applications. From a technical perspective, this article examines the different classes of pulse sequences currently available to image spins with ultra-short transverse relaxation times (T2 and T2*), with particular attention focused on the newest member of the class, sweep imaging with Fourier transformation (SWIFT).


HPV-18 E2^E4 chimera: 2 new spliced transcripts and proteins induced by keratinocyte differentiation.
The Human Papillomavirus (HPV) E4 is known to be synthesized as an E1^E4 fusion resulting from splice donor and acceptor sites conserved across HPV types. Here we demonstrate the existence of 2 HPV-18 E2^E4 transcripts resulting from 2 splice donor sites in the 5' part of E2, while the splice acceptor site is the one used for E1^E4. Both E2^E4 transcripts are up-regulated by keratinocyte differentiation in vitro and can be detected in clinical samples containing low-grade HPV-18-positive cells from Pap smears. They give rise to two fusion proteins in vitro, E2^E4-S and E2^E4-L. Whereas we could not differentiate E2^E4-S from E1^E4 in vivo, E2^E4-L could be formally identified as a 23 kDa protein in raft cultures in which the corresponding transcript was also found, and in a biopsy from a patient with cervical intraepithelial neoplasia stage I-II (CINI-II) associated with HPV-18, demonstrating the physiological relevance of E2^E4 products.


Predictors of retained hemothorax after trauma and impact on patient outcomes.
Hemo/pneumothoraces are a common result of thoracic injury. Some of these injuries will be complicated by retained hemothorax (RH), which has previously been shown to be associated with longer hospitalizations. It has been proposed that early versus delayed intervention with video-assisted thoracoscopic surgery can reduce the duration of mechanical ventilation, hospital and ICU LOS, and costs in patients with RH. However, little is known regarding the effect of RH on these outcomes relative to patients with uncomplicated hemo/pneumothoraces. The aim of our study was to characterize factors present on admission that may be associated with RH and assess the impact of RH on outcomes. A retrospective chart review was conducted and included all patients who underwent tube thoracostomy (TT) for traumatic hemo/pneumothorax admitted to a single urban adult and pediatric level I trauma center from January 2008 to September 2013. The study cohort included 398 patients, 17.6 % developed RH. RH was associated with significantly longer total duration of TT drainage (p < 0.001), hospital LOS (p < 0.001), and total hospital charges (p < 0.001). These associations remained significant in a subgroup analysis excluding patients with traumatic brain injury. Patients with bilateral injuries (OR 4.25, p < 0.001) and patients intubated on the day of admission (OR 2.30, p = 0.002) were significantly more likely to develop RH. There was also a small, but highly significant, association between increasing ISS and the development of RH (OR 1.07, p < 0.001). Our study suggests patients requiring ventilator support on admission and those with bilateral injuries are at increased risk of developing RH. Early identification of patients at risk for RH may allow for earlier intervention and potential benefits to the patient.


Comparative studies of the dorsal surface of the tongue in three mammalian species by scanning electron microscopy.
Comparative features of the dorsal tongue epithelia in musk shrews, mongooses and rats were described. The shapes of the filiform papillae were different in each of the species. The distribution pattern of filiform papillae was similar both in the musk shrews and mongoose, in that the form of filiform papillae changed gradually from the lingual apex to the posterior part of the lingual body. By contrast, the different types of filiform papillae were distributed on definite areas of the dorsal lingual surface in the rat. Microridges on the interpapillar surface in the musk shrew and mongoose presented a clear outline, but those of the rat were not so distinct. In all species, the upper surface of filiform papillae did not show any distinct microridges.


After-effects of stochastic synaptic Renshaw cell excitation on their discharge probability.
We have studied Renshaw cell (RC) responses to pseudo-Poisson stimulus sequences (small intervals below 5 ms excluded; mean rates between 10 and 45 pulses/s) delivered to motor axons in ventral roots or muscle nerves. Average RC responses to stimuli in a single stimulus channel depended upon the preceding stimulation history. The responses to pairs of stimuli separated by variable intervals (irrespective of intervening stimuli) generally showed a long depressant effect (from 25 to more than 250 ms) of the first ('conditioning') stimulus; in a number of cases a briefer facilitating effect preceded the depressant action. Several possible causes of these conditioning effects are discussed.


A testicular seminoma in a rabbit.
The histological, ultrastructural and biological features are described of a spontaneously occurring seminoma in the testis of a male domestic rabbit. The tumour was structurally similar to seminomas in other animal species.


Relationship between different measures of pain-related fear and physical capacity of the spine in patients with chronic low back pain.
It has been controversially stated that pain-related fear is a more important determining factor for disability in chronic low back pain (CLBP) than pain or physical impairment in itself. So far, the relationship between psychological and physiological determinants of chronic pain, that is, pain-related fear and physiological abilities, remains unclear. To evaluate whether pain-related fear assessed by different tools (both task specific and non task specific) is related to physical capacity measured by specific spine tests and, secondarily, to explore the relationship between different pain-related fear assessment tools. Cross-sectional study. Fifty patients with CLBP (50% women; meanage [standard deviationage]: 44.2 [9.5 years]). Physical capacity by means of three specific spine tests, that is, the finger-floor distance test (flexibility), a maximal isometric strength test of trunk extensor muscles (strength), and the Sorensen test (endurance). Pain-related fear by means of self-report measures, that is, the Tampa Scale for Kinesiophobia (TSK), the Photograph Series of Daily Activities (PHODA), and a fear visual analog scale (FVAS) tailored to the spine tests. Participants were asked to complete the TSK and PHODA and to perform the three spine tests. Right before performing each of the spine tests, an FVAS was filled out. Linear regression analyses controlling for gender and age were performed to study the association between the pain-related fear measurements and the results of the spine tests. To investigate the relationship between the pain-related fear measurements, correlation tests were performed. The linear regression analyses revealed that neither the TSK and PHODA scores nor the FVAS scores were significantly related to the physical capacity measurements. The correlational tests showed no significant correlation between the PHODA, TSK, and FVAS scores. The present study shows that neither the task-specific tool (FVAS) nor the non task-specific questionnaires (TSK and PHODA) were significantly correlated to the spine tests in patients with CLBP. This is contrary to earlier evidence according to which physical capacity is inversely related to the level of pain-related fear, and it suggests that one should not draw conclusions about physical capacity based on pain-related fear scores. Furthermore, the different assessment tools for pain-related fear were surprisingly not correlated with each other.


A p38 mitogen-activated protein kinase inhibitor attenuates cardiotonic steroids-induced apoptotic and stress signaling in a Sw-71 cytotrophoblast cell line.
Preeclampsia (preE) is characterized by abnormal placentation. Marinobufagenin (MBG), a cardiotonic steroid (CTS), inhibits the function of cytotrophoblast cells (CTBs). We demonstrated that CTSs induce anti-angiogenic and anti-proliferative effects in Sw-71 CTBs. This study tests that CTSs induce apoptotic and stress signaling. Human extravillous Sw-71 CTBs were incubated with 0, 0.1, 1, 10, and 100 nM of each of three CTSs (MBG, cinobufatalin (CINO) and ouabain (OUB)) for 48 h. Some cells were pretreated with 10 μM p38 mitogen-activated protein kinase (p38 MAPK) inhibitor (SB203580) for 2 h prior to CTSs treatment. We analyzed p38 MAPK phosphorylation, expression of pro-inflammatory protein cyclooxygenase-2 (Cox-2) and ratio of pro-apoptotic Bcl-2-associated X protein (Bax) to anti-apoptotic Bcl-2 protein by western blot in CTSs-treated CTBs lysates. Levels of vascular endothelial growth factor (VEGF), placental growth factor (PlGF), soluble fms-like tyrosine kinase-1 (sFlt-1) and soluble endoglin (sEng) were measured in culture media using ELISA kits. Statistical comparisons were performed using analysis of variance with Duncan's post hoc test. p38 MAPK phosphorylation, expression of Cox-2 and Bax/Bcl-2 was upregulated (*p < 0.05) in CTBs exposed to ≥ 0.1 nM CTSs. Secretion of sFlt-1 and sEng were increased while VEGF and PIGF were decreased in Sw-71 CTBs treated ≥1 nM of each CTSs (*p < 0.01 for each). The SB203580 pretreatment of CTBs significantly attenuated CTS-induced effects. Exposure of Sw-71 CTBs to CTSs induced apoptotic and stress signaling and causing anti-angiongenic effect. The observed diminution of CTS-induced signaling by SB203580 pretreatment implicates p38 MAPK as a regulator of these pathways.


Intestinal tract injury by drugs: Importance of metabolite delivery by yellow bile road.
Drug secretion into bile is typically considered a safe route of clearance. However, biliary delivery of some drugs or their reactive metabolites to the intestinal tract evokes adverse consequences due to direct toxic actions or indirect disruption of intestinal homeostasis. Biliary concentration of the chemotherapy agent 5-fluorodeoxyuridine (FUDR) and other compounds is associated with bile duct damage while enterohepatic cycling of antibiotics contributes to the disruptions of gut flora that produce diarrhea. The goal of this review is to describe key evidence that biliary delivery is an important factor in the intestinal injury caused by representative drugs. Emphasis will be given to 3 widely used drugs whose reactive metabolites are plausible causes of small intestinal injury, namely the nonsteroidal anti-inflammatory drug (NSAID) diclofenac, the immunosuppressant mycophenolic acid (MPA), and the chemotherapy agent irinotecan. Capsule endoscopy and other sensitive diagnostic techniques have documented a previously unappreciated, high prevalence of small intestinal injury among NSAID users. Clinical use of MPA and irinotecan is frequently associated such severe intestinal injury that dosage must be reduced. Observations from clinical and experimental studies have defined key events in the pathogenesis of these drugs, including roles for multidrug resistance-associated protein 2 (MRP2) and other transporters in biliary secretion and adduction of enterocyte proteins by reactive acyl glucuronide metabolites as a likely mechanism for intestinal injury. New strategies for minimizing the adverse intestinal consequences of irinotecan chemotherapy illustrate how basic information about key events in the biliary secretion of drugs and the nature of their proximate toxicants can lead to safer protocols for drugs.


Annual and intradiurnal variation of dominant airborne pollen and the effects of meteorological factors in Çeşme (Izmir, Turkey).
In this present study, airborne pollen in Çeşme was investigated between February 17, 2012 and February 17, 2014 using the volumetric method. Çeşme, one of Turkey's most important tourism centers, which attracts numerous local and foreign tourists each year, is a district of Izmir, a province in the western part of Turkey. During the 2-year study, 12,905 pollen grains belonging to 64 taxa (33 arboreal, 31 non-arboreal plants) were detected. However, the 2-year data results revealed that the taxa with the pollen concentration more than 4% in the atmosphere were Cupressaceae/Taxaceae (4268 pollen, 33.07%), Olea europaea (1614 pollen grains, 12.51%), Pinaceae (1085 pollen grains, 8.41%), Quercus spp. (1081 pollen grains, 8.38%), Pistacia spp. (743 pollen grains, 5.76%), and Poaceae (557 pollen grains, 4.32%), all of which comprised 72.44% of the total count. The relationship between the daily pollen counts belonging to these six taxa and the hourly average temperature (°C), daily precipitation (mm), relative humidity (%), and wind speed (km/h) was assessed using the Spearman correlation test, and significant results were determined. During the study, the intradiurnal distribution of the aforementioned pollen varied. The highest pollen concentration was detected between 11:00 a.m. and 6:00 p.m. (first year 30.3%; second year 30.1%).


Functional cell-based uHTS in chemical genomic drug discovery.
The availability of genomic information significantly increases the number of potential targets available for drug discovery, although the function of many targets and their relationship to disease is unknown. In a chemical genomic research approach, ultra-high throughput screening (uHTS) of genomic targets takes place early in the drug discovery process, before target validation. Target-selective modulators then provide drug leads and pharmacological research tools to validate target function. Effective implementation of a chemical genomic strategy requires assays that can perform uHTS for large numbers of genomic targets. Cell-based functional assays are capable of the uHTS throughput required for chemical genomic research, and their functional nature provides distinct advantages over ligand-binding assays in the identification of target-selective modulators.


Familial androgenetic alopecia in siblings with normal endocrinological status.
Androgenetic alopecia (AGA) is the most common type of hair loss in adults, but it has been rarely reported in children. We report two cases of AGA in two siblings, aged 6 and 8, whose mother had the same condition, without any association with other skin diseases or abnormalities in endocrinologic examinations.


Cyclic adenosine monophosphate regulates vasoactive intestinal polypeptide and enkephalin biosynthesis in cultured bovine chromaffin cells.
When bovine chromaffin cells are dissociated from the adult adrenal gland and placed in culture, they begin to synthesize vasoactive intestinal polypeptide (VIP); they also contain high levels of enkephalin peptides. The regulation of expression of VIP and enkephalin in these cells by cyclic nucleotides was examined. Exposure of cultured chromaffin cells to cholera toxin (CT), forskolin (F), isobutylmethylxanthine (IBMX), 8-bromo-cyclic AMP (8-Br-cAMP) or dibutyryl cyclic AMP (dbcAMP) increases VIP and met-enkephalin biosynthesis as measured by an increase in total (cellular + secreted) VIP and met-enkephalin in treated versus untreated cells. Enkephalin levels increase 1.5 to 3.0 -fold while VIP levels increase 10 to 20 -fold after 48 hours of exposure to the compounds above. Increased enkephalin levels are maximum by 48 hours of exposure; VIP levels are elevated by forskolin or cholera toxin already at 17 hours of exposure. Neither forskolin nor cholera toxin alter catecholamine levels in cultured cells even after 72 hours of exposure. Thus, VIP and ME expression in cultured chromaffin cells can be regulated by intracellular cAMP. Measurement of preproenkephalin messenger RNA (mRNAenk) by quantitative Northern blot hybridization with a preproenkephalin complementary DNA probe revealed that exposure of chromaffin cells to forskolin elicits a greater than two-fold increase in mRNAenk, suggesting that the regulation of neuropeptide expression by elevated intracellular cAMP occurs at a transcriptional locus.


Patulin biosynthesis: enzymatic and nonenzymatic transformations of the mycotoxin (E)-ascladiol.
A mycotoxin, (E)-ascladiol, was established as a direct precursor of patulin in cell-free preparations of Penicillium urticae patulin-minus mutants J1 and S11, but not S15. Isomerization to a side product, (Z)-ascladiol, was nonenzymatically catalyzed by sulfhydryl compounds.


Primary and Metastatic Bone Tumors of the Patella: Literature Review and Institutional Experience.
Patellar tumors are rare but certainly must be considered in the differential diagnosis in patients with knee pain. Diagnosis can be challenging as often patellar neoplasms are confused with benign conditions and their clinical presentation is usually not specific. We performed an institutional and a literature review to determine what are the most common tumors affecting the patella and what is the best management. This is a case series from our institution including all patients with benign, malignant, and metastatic patellar neoplasms. Charts were reviewed for patient demographics, clinical presentation, pathology characteristics, radiographic classification, and oncologic and functional outcomes. Twenty-four patients were identified; twelve patients had benign lesions, 10 metastatic and 2 primary malignant tumors. Chondroblastoma and Giant Cell Tumor were the most common tumors. Management of benign lesions with intralesional curettage and packing with bone graft or cement demonstrated excellent results with no local recurrence. In terms of malignant tumors, the spectrum of treatment is variable; it could range from medical management alone or in combination with surgical procedures to total patellectomy with reconstruction of the extensor mechanism. Patellar tumors should be part of the differential in patients with chronic knee pain that does not respond to initial conservative interventions. Recurrence rate with intralesional curettage and bone grafting or cement packing is very low and therefore should be the treatment of choice for benign intraosseous neoplasms. Resection with negative margins in malignant neoplasms or bone metastasis decreases local recurrence but only in the former group there is a potential impact in survival.


Membrane-associated, fucose-containing glycoproteins and glycolipids of cultured epithelial cells from human colonic adenocarcinoma and fetal intestine.
Fucose-containing glycoproteins and glycolipids were compared in three human colon cancer cell lines and five human fetal intestinal epithelial cell lines. Cells were labeled by culturing cells in the presence of L-[3H]-fucose. Fucose was incorporated into both the membrane and cytoplasmic fractions of all three colon cancer cell lines to a much lesser extent than into fetal cells. When the relative fucose labeling of glycolipids and glycoproteins were examined, a much greater proportion of fucose labeling in the membrane was associated with lipid in colon cancer cells (11.6-16.7%) compared to fetal intestinal cells (1.3-2.5%). Fluorographic analysis of SDS-polyacrylamide gel electrophoresis of fucose-labeled glycoproteins revealed a rather uniform labeling pattern of fetal intestinal cells which was distinct from those of colon cancer cells. Thin-layer chromatographic analysis of fucose labeled glycolipids of all three colon cancer cell lines indicated the presence of fucose-containing glycolipids with carbohydrate chain lengths greater than five sugars. Glycolipids of the SKCO-1 cells in particular appear to consist predominantly of complex fucose-containing glycolipids. These results indicate that significant qualitative differences in the fucose-containing glycoproteins and glycolipids exist between the membranes of human colon cancer cells and fetal intestinal cells and that complex fuco-glycolipids with long carbohydrate side chains are present in the three human colon cancer cell lines.


Phenamacril and carbendazim regulate trichothecene mycotoxin synthesis by affecting ROS levels in F. asiaticum.
Fusarium head blight caused by Fusarium asiaticum is an important cereal crop disease, and the trichothecene mycotoxins produced by F. asiaticum can contaminate wheat grain, which is very harmful to humans and animals. To effectively control FHB in large areas, the application of fungicides is the major strategy; however, the application of different types of fungicides has varying influences on the accumulation of trichothecene mycotoxins in F. asiaticum. In this study, phenamacril inhibited trichothecene mycotoxin accumulation in F. asiaticum; however, carbendazim (N-1H-benzimidazol-2-yl-carbamic acid, methyl ester) induced trichothecene mycotoxin accumulation. Additionally, phenamacril led to a lower level of reactive oxygen species (ROS) by inducing gene expression of the catalase and superoxide dismutase (SOD) pathways in F. asiaticum, whereas carbendazim stimulated ROS accumulation by inhibiting gene expression of the catalase and SOD pathways. Based on these results, we conclude that phenamacril and carbendazim regulate trichothecene mycotoxin synthesis by affecting ROS levels in F. asiaticum.


Identification of gallotannins and ellagitannins in aged wine spirits: A new perspective using alternative ageing technology and high-resolution mass spectrometry.
This research was focused on identifying gallotannins and ellagitannins degradation pathways to better understand their behavior in complex media such as wine spirits (WS). A WS was aged with chestnut wood staves with three levels of micro-oxygenation, nitrogen, and using wooden barrels. Gallotannins and ellagitannins were identified by LC-ESI-HRMS/MS using a Q-TOF in samples collected at 8, 21, 60, 180, 270, and 365 days of ageing, allowed comparing their relative abundances according to the ageing technology. It was established for the first time, the importance of oxygen in gallotannins and ellagitannins formation/degradation pathways in WS and shading light into the explanation for the steady increase of gallic and ellagic acid contents on WS during ageing. The results also highlighted the presence of penta-O-galloyl-β-d-glucose, tetra-O-galloyl-β-d-glucose, tri-O-galloyl-β-d-glucose, di-O-galloyl-β-d-glucose, and mono-O-galloyl-β-d-glucose, 2,3-(S)-hexahydroxydiphenoyl-β-d-glucose, pedunculagin, isomers vescalagin/castalagin and two products stemming from ethanol-promoted oxidation of castalagin/vescalagin and vescalin/castalin, in the composition WS aged with chestnut wood.


Dating the demise: neandertal extinction and the establishment of modern humans in the southern Caucasus.
This paper considers the recent radiometric dating (14C-AMS, TL, ESR) of 76 late Middle and early Upper Paleolithic samples from Ortvale Klde Rockshelter, located in the Republic of Georgia. We present a critical evaluation of each date based on its stratigraphic and archaeological context, its pretreatment and contamination history, and its resulting accuracy and precision, the goal being to establish a sound chronology for the site. Only by systematically identifying aberrant dates within a data set and isolating them from further analysis can we hope to understand cultural and biological phenomena on an accurate temporal scale. Based on the strict discard protocol outlined here, we omit 25% of the dated samples from the analysis. The remaining data speak to the lengthy tenure of Neandertals in the region, but also to their relatively rapid demise and the establishment of modern human populations approximately 38-34 ka 14C BP (42-39 kacalBP(Hulu)). We compare these chronometric data with those from the neighboring sites of Bronze and Dzudzuana caves, as well as Mezmaiskaya Cave, located in the northern Caucasus. While the lack of key contextual information limit our ability to subject these other data sets to the same critical evaluation procedure, they provide the first interregional temporal assessment of the Middle to Upper Paleolithic "transition," the results of which suggest an initial expansion of modern humans into the southern Caucasus followed by expansion along the Black Sea coast and into the northern Caucasus.


Circulating vitamin C and digestive system cancers: Mendelian randomization study.
Vitamin C is an antioxidant with a potential role in the prevention of digestive system cancers, but there is yet no consensus whether vitamin C has a causal role in these cancers. The aim of this study was to utilize Mendelian randomization to decipher the potential causal associations of vitamin C with risk of digestive system cancers. Ten genetic variants previously found to be significantly associated with circulating vitamin C were used as instrumental variables. Effect size estimates for the genetic associations of the vitamin C-associated genetic variants with six major malignancies of digestive system were obtained from the FinnGen (N = 309 154) and UK Biobank (N = 367 542) studies. Results from the two studies were combined using meta-analysis. Genetically predicted higher circulating vitamin C showed a suggestive association with lower risk of small intestine and colorectal cancer after accounting for multiple testing. The odds ratio per 1 standard deviation increment in circulating vitamin C was 0.55 (95% confidence interval 0.32-0.94; P = 0.029) for small intestine cancer and 0.84 (95% confidence interval 0.73-0.96; P = 0.013) for colorectal cancer. There was a suggestive association between genetically predicted higher circulating vitamin C with lower risk of liver cancer in FinnGen but no association in the meta-analysis (odds ratio 0.69; 95% CI 0.36-1.32; P = 0.265). Genetically predicted circulating vitamin C was not associated with cancers of the esophagus, stomach, or pancreas. This Mendelian randomization study indicates that vitamin C might play a role in the prevention of small intestine and colorectal cancer.


Advanced age is an independent prognostic factor of disease progression in high-risk prostate cancer: results in 180 patients treated with robot-assisted radical prostatectomy and extended pelvic lymph node dissection in a tertiary referral center.
This study aimed to assess more clinical and pathological factors associated with prostate cancer (PCa) progression in high-risk PCa patients treated primarily with robot-assisted radical prostatectomy (RARP) and extended pelvic lymph node dissection (ePLND) in a tertiary referral center. In a period ranging from January 2013 to October 2020, RARP and ePLND were performed on 180 high-risk patients at Azienda Ospedaliera Universitaria Integrata of Verona (Italy). PCa progression was defined as biochemical recurrence/persistence and/or local recurrence and/or distant metastases. Statistical methods evaluated study endpoints, including Cox's proportional hazards, Kaplan-Meyer survival curves, and binomial logistic regression models. The median age of included patients was 66.5 [62-71] years. Disease progression occurred in 55 patients (30.6%), who were more likely to have advanced age, palpable tumors, and unfavorable pathologic features, including high tumor grade, stage, and pelvic lymph node invasion (PLNI). On multivariate analysis, PCa progression was predicted by advanced age (≥ 70 years) (HR = 2.183; 95% CI = 1.089-4377, p = 0.028), palpable tumors (HR = 3.113; 95% CI = 1.499-6.465), p = 0.002), and PLNI (HR = 2.945; 95% CI = 1.441-6.018, p = 0.003), which were associated with clinical standard factors defining high-risk PCa. Age had a negative prognostic impact on elderly patients, who were less likely to have palpable tumors but more likely to have high-grade tumors. High-risk PCa progression was independently predicted by advanced age, palpable tumors, and PLNI, which is associated with standard clinical prognostic factors. Consequently, with increasing age, the prognosis is worse in elderly patients, who represent an unfavorable age group that needs extensive counseling for appropriate and personalized management decisions.


Insurance coverage and financial burden for families of children with special health care needs.
To examine the role of insurance coverage in protecting families of children with special health care needs (CSHCN) from the financial burden associated with care. Data from the 2001 National Survey of Children with Special Health Care Needs were analyzed. We built 2 multivariate regression models by using "work loss/cut back" and "experiencing financial problems" as the dependent variables, and insurance status as the primary independent variable of interest while adjusting for income, race/ethnicity, functional limitation/severity, and other sociodemographic predictors. Approximately 29.9% of CSHCN live in families where their condition led parents to report cutting back on work or stopping work completely. Families of 20.9% of CSHCN reported experiencing financial difficulties due to the child's condition. Insurance coverage significantly reduced the likelihood of financial problems for families at every income level. The proportion of families experiencing financial problems was reduced from 35.7% to 23.0% for the poor and 44.9% to 24.5% for low-income families with continuous insurance coverage (P < .01 for both comparisons). Similarly, the proportion of parents having to cut back or stop work was reduced from 42.8% to 35.9% for the poor (P < .05) and 43.5% to 33.9% for low-income families (P < .01). Continuous health insurance coverage provides protection from financial burden and hardship for families of CSHCN in all income groups. This evidence is supportive of policies designed to promote universal coverage for CSHCN. However, many poor and low-income families continue to experience work loss and financial problems despite insurance coverage. Hence, health insurance should not be viewed as a solution in itself, but instead as one element of a comprehensive strategy to provide financial safety for families with CSHCN.


Paeonol, the active component of Cynanchum paniculatum, ameliorated schizophrenia-like behaviors by regulating the PI3K-Akt-GSK3β-NF-κB signalling pathway in MK-801-treated mice.
Cynanchum paniculatum (Bunge) Kitag. ex H. Hara (Asclepiadaceae) have been traditionally used in East Asia as analgesic or antiviral agents. Interestingly, some Chinese and Korean traditional medicinal books reported that the use of C. paniculatum in the treatment of psychotic symptoms, such as hallucinations and delusions. In this study, we aimed to investigate whether C. paniculatum could improve sensorimotor gating disruption in mice with MK-801-induced schizophrenia-like behaviors. We also aimed to identify the active component of C. paniculatum that could potentially serve as a treatment for schizophrenia and found that paeonol, the major constituent compound of C. paniculatum, showed potential as a treatment for schizophrenia. To assess the effect of paeonol on mice with MK-801-induced schizophrenia-like behaviors, we carried out a series of behavioral tests related with symptoms of schizophrenia. In addition, we utilized Western blotting and ELISA techniques to investigate the antipsychotic actions of paeonol. C. paniculatum extract (100 or 300 mg/kg) and paenol (10 or 30 mg/kg) significantly reversed MK-801-induced prepulse deficits in acoustic startle response test. In addition, paeonol (10 or 30 mg/kg) attenuated social novelty preference and novel object recognition memory on MK-801-induced schizophrenia-like behaviour in mice. Furthermore, the phosphorylation levels of PI3K, Akt, GSK3β and NF-κB, as well as related pro-inflammatory cytokine, such as IL-1β and TNF-α, were significantly reversed by the administration of paeonol (10 or 30 mg/kg) in the prefrontal cortex of MK-801-treated mice. Collectively, these data show that paeonol can potentially be used as an agent for treating sensorimotor gating deficits, negative symptoms, and cognitive deficits, such as those observed in schizophrenia with few adverse effects.


High resolution CryoFESEM of microbial surfaces.
The outer surfaces of three microorganisms, Giardia lamblia, Enterococcus faecalis, and Proteus mirabilis, were investigated by cryo-immobilization followed by sublimation of extracellular ice and cryocoating with either Pt alone or Pt plus carbon. Cryocoated samples were examined at -125 degrees C in either an in-lens field emission SEM or a below-the-lens field emission SEM. Cryocoating with Pt alone was sufficient for low magnification observation, but attempts to do high-resolution imaging resulted in radiolysis and cracking of the specimen surface. Double coating with Pt and carbon, in combination with high resolution backscatter electron detectors, enabled high-resolution imaging of the glycocalyx of bacteria, revealing a sponge-like network over the surface. High resolution examination of bacterial flagella also revealed a periodic substructure. Common artifacts included radiolysis leading to "cracking" of the surface, and insufficient deposition of Pt resulting in the absence of detectable surface topography.


Reduced quantitative ultrasound bone mineral density in HIV-infected patients on antiretroviral therapy in Senegal.
Bone status in HIV-infected patients on antiretroviral treatment (ART) is poorly documented in resource-limited settings. We compared bone mineral density between HIV-infected patients and control subjects from Dakar, Senegal. A total of 207 (134 women and 73 men) HIV-infected patients from an observational cohort in Dakar (ANRS 1215) and 207 age- and sex-matched controls from the general population were enrolled. Bone mineral density was assessed by quantitative ultrasound (QUS) at the calcaneus, an alternative to the reference method (i.e. dual X-absorptiometry), often not available in resource-limited countries. Mean age was 47.0 (±8.5) years. Patients had received ART for a median duration of 8.8 years; 45% received a protease inhibitor and 27% tenofovir; 84% had undetectable viral load. Patients had lower body mass index (BMI) than controls (23 versus 26 kg/m(2), P<0.001). In unadjusted analysis, QUS bone mineral density was lower in HIV-infected patients than in controls (difference: -0.36 standard deviation, 95% confidence interval (CI): -0.59;-0.12, P = 0.003). Adjusting for BMI, physical activity, smoking and calcium intake attenuated the difference (-0.27, CI: -0.53;-0.002, P = 0.05). Differences in BMI between patients and controls explained a third of the difference in QUS bone mineral density. Among patients, BMI was independently associated with QUS bone mineral density (P<0.001). An association between undetectable viral load and QUS bone density was also suggested (β = 0.48, CI: 0.02;0.93; P = 0.04). No association between protease inhibitor or tenofovir use and QUS bone mineral density was found. Senegalese HIV-infected patients had reduced QUS bone mineral density in comparison with control subjects, in part related to their lower BMI. Further investigation is needed to clarify the clinical significance of these observations.


Meta-tyrosine modulates the immune response induced by bacterial endotoxins.
Sepsis is characterized by an early pro-inflammatory phase followed by compensatory anti-inflammatory mechanisms that lead to a late generalized immunosuppression, period where most deaths occur. Immunotherapy approaches to recover the immunocompetence in sepsis are similar to those used in cancer. Meta-tyrosine (m-Tyr) is a product of oxidative stress present in circulation during the sepsis and cancer-associated pro-inflammatory stages. In this work, considering its potential participation in pro-inflammatory processes, we evaluate the effect of m-Tyr during LPS induced immunosuppression phase in a murine model. In addition, we examine the effect of m-Tyr in a vaccination strategy using a weakly immunogenic tumor model. Our results showed that m-Tyr could prevent the establishment of immunosuppression and rescue the host from an installed immunosuppression induced by LPS. These effects were parallel to the ability of m-Tyr to improve the pro-inflammatory effects induced by LPS and inhibit the anti-inflammatory action of dexamethasone. Also, m-Tyr treatment prevents both the reduction of splenic lymphocytes and the increase of the expression of programmed death ligand-1 in splenic myeloid cells associated with immunosuppression. Besides, treatment with m-Tyr increased the protective effect of an anti-tumor vaccine, suggesting that m-Tyr could improve the immune response. In summary, we suggest that m-Tyr can modulate critical immunological indicators through the inflammatory context, which could improve the management of diseases, such as sepsis and cancer, in which immunosuppression is a significant clinical problem.


Flow dynamics and energy efficiency of flow in the left ventricle during myocardial infarction.
Cardiovascular disease is a leading cause of death worldwide, where myocardial infarction (MI) is a major category. After infarction, the heart has difficulty providing sufficient energy for circulation, and thus, understanding the heart's energy efficiency is important. We induced MI in a porcine animal model via circumflex ligation and acquired multiple-slice cine magnetic resonance (MR) images in a longitudinal manner-before infarction, and 1 week (acute) and 4 weeks (chronic) after infarction. Computational fluid dynamic simulations were performed based on MR images to obtain detailed fluid dynamics and energy dynamics of the left ventricles. Results showed that energy efficiency flow through the heart decreased at the acute time point. Since the heart was observed to experience changes in heart rate, stroke volume and chamber size over the two post-infarction time points, simulations were performed to test the effect of each of the three parameters. Increasing heart rate and stroke volume were found to significantly decrease flow energy efficiency, but the effect of chamber size was inconsistent. Strong complex interplay was observed between the three parameters, necessitating the use of non-dimensional parameterization to characterize flow energy efficiency. The ratio of Reynolds to Strouhal number, which is a form of Womersley number, was found to be the most effective non-dimensional parameter to represent energy efficiency of flow in the heart. We believe that this non-dimensional number can be computed for clinical cases via ultrasound and hypothesize that it can serve as a biomarker for clinical evaluations.


Electrophysiologic and pharmacokinetic profile of the new antiarrhythmic drug TYB-3823 in humans.
In an open-label study, electrophysiology and pharmacokinetics of TYB-3823, a new antiarrhythmic compound, were investigated. Sixteen patients underwent an electrophysiologic study before and after intravenous (i.v.) administration of TYB-3823. Two patients each received the following increasing doses: 0.2, 0.4, 0.8, and 1 mg/kg. Eight patients received 1.2-mg/kg. TYB-3823 concentrations followed a biexponential decrease with a terminal half-life (t1/2) of 3.88 +/- 0.87 h. Clearance was 47.2 +/- 18.5 L/h, and volume of distribution was 250 +/- 77 L. Dose-dependent pharmacokinetics were evident. Significant effects of TYB-3823 were apparent at doses > or = 0.8 mg/kg (n = 12), including increase in the AH and HV interval from 92 +/- 17 to 105 +/- 19 ms (p < 0.002) and 47 +/- 7 to 60 +/- 12 ms (p < 0.005), respectively. QRS duration was prolonged from 100 +/- 16 to 126 +/- 22 ms (p < 0.001), accompanied by an increase of the corrected QT interval from 425 +/- 28 to 465 +/- 37 ms (p < 0.002). The corrected JT interval remained unchanged, however, refractoriness did not change, but monophasic action potential duration (APD) tended to decrease. TYB-3823 appeared effective against reinduction of all arrhythmias observed during the control study [atrial fibrillation, atrioventricular (AV) nodal tachycardias]. TYB-3823 depresses conduction velocity significantly without prolonging refractoriness. Therefore, TYB-3823 may be classified as a class 1C antiarrhythmic. On the basis of the present results, additional class 1B activity cannot be excluded. TYB-3823 has antiarrhythmic properties, appears to be devoid of proarrhythmic effects, and is well tolerated.


Reproductive responses of early postpartum dairy cattle to continuous treatment with a GnRH agonist (deslorelin) for 28 days to delay the resumption of ovulation.
An experiment was conducted to investigate the potential of chronic delivery of a potent GnRH agonist (deslorelin) via subcutaneous implants to delay the resumption of ovulatory cycles in postpartum dairy cattle. Cows received either a single deslorelin implant (n=40; DES) within 7 days of calving or were untreated (n=24; CON). Blood samples were collected thrice weekly during the period the implants were in place. Plasma concentrations of progesterone (P4) and 17beta-oestradiol (E2) were measured along with selected serum metabolites. Implants were removed after 28 days and cattle monitored daily for behavioral oestrus. Serial weekly blood samples were collected to detect the occurrence of ovulation. Cows were artificially inseminated as they were detected in oestrus from 30 days after implant removal. Pregnancy status was subsequently determined by manual palpation of uterine contents at strategic intervals. Insertion of implants induced ovulation in 3/40 cows as determined by a rise in progesterone 7 days later. Deslorelin implants delayed the onset of ovulatory cycles compared with untreated herdmates (mean 43.4+/-4.2 versus 57.3+/-1.6 days postpartum; P<0.001). A noticeable delay of at least 12 days was observed between implant removal and the first animals ovulating. Mean plasma E2 concentrations during the period the implants were in place were similar for DES and CON cows that experienced a prolonged spontaneous postpartum anoestrus (low P4 >60 days), although both groups had concentrations only 20% of CON cows that had ovulated prior to 30 days postpartum. The patterns of recovery following implant removal were highly variable. A number of DES cows showed a low and transient rise in plasma progesterone around 21 days after implant removal. Some cows displayed oestrus but did not appear to form a fully functional corpus luteum with this phenomenon being more prevalent among DES cows (7 of 37 versus 1 of 21; P<0.05). Overall, significantly more DES cows were detected in oestrus without ovulating compared to CON cows. Final pregnancy rates did not differ between DES and CON groups. The mean time to conception for DES cows was longer (21.2+/-5.6 versus 41.1+/-7.4 days, CON versus DES; P<0.01). This difference was not present if the time from first ovulation to conception was compared (50.5+/-5.3 versus 43.5+/-9.3 days, CON versus DES; P>0.05). Deslorelin implants provided a reliable method of inducing anoestrus when treatment was initiated prior to 3 days postpartum. A variable pattern of recovery was observed which delayed conception but did not ultimately reduce the final proportion pregnant at the completion of mating. The study demonstrates the potential of GnRH agonists to control postpartum reproductive function to manipulate the fertility of dairy cows.


Assessment of DNA damage and mRNA/miRNA transcriptional expression profiles in hyperglycemic versus non-hyperglycemic patients with type 2 diabetes mellitus.
The development of type 2 diabetes mellitus (T2D) is associated with a number of genetic and environmental factors. Hyperglycemia, a T2D hallmark, is related to several metabolic complications, comorbidities and increased DNA damage. However, the molecular alterations of a proper glucose control are still unclarified. In this study, we aimed to evaluate DNA damage (comet assay), as well as to compare the transcriptional expression (mRNA and miRNA analyzed by the microarray technique) displayed by peripheral blood mononuclear cells (PBMCs) from three distinct groups: hyperglycemic T2D patients (T2D-H, n=14), non-hyperglycemic T2D patients (T2D-N, n=15), and healthy non-diabetic individuals (n=16). The comet assay revealed significantly (p<0.05) higher levels of DNA damage in T2D-H group compared to both T2D-N and control groups, while a significant difference was not observed between the control and T2D-N groups. After bioinformatics analysis, the differentially expressed mRNAs were subjected to functional enrichment analysis (DAVID) and inflammatory response was among the enriched terms found when comparing T2D-N with controls and T2D-H with T2D-N. Concerning the gene set enrichment and gene set analyses, among the differentially expressed gene sets, three were of interest: regulation of DNA repair (T2D-H versus T2D-N), superoxide response (T2D-H versus control group), and response to endoplasmic reticulum stress (T2D-H versus control group). We also identified miRNAs related with T2D and hyperglycemia not yet associated with these conditions in the literature. Some of the differentially expressed mRNAs were among the predicted targets of the differentially expressed miRNAs. Our results showed the association of hyperglycemia with increased DNA damage and aberrant expression of miRNAs and genes related to several biological processes, such as inflammation, DNA repair, ROS production and antioxidant defense, highlighting the importance of proper glycemic control. Moreover, the transcriptional expression of miRNAs provided novel information for understanding the regulatory mechanisms involved in the T2D progression.


Custom fit 3D-printed brain holders for comparison of histology with MRI in marmosets.
MRI has the advantage of sampling large areas of tissue and locating areas of interest in 3D space in both living and ex vivo systems, whereas histology has the ability to examine thin slices of ex vivo tissue with high detail and specificity. Although both are valuable tools, it is currently difficult to make high-precision comparisons between MRI and histology due to large differences inherent to the techniques. A method combining the advantages would be an asset to understanding the pathological correlates of MRI. 3D-printed brain holders were used to maintain marmoset brains in the same orientation during acquisition of ex vivo MRI and pathologic cutting of the tissue. The results of maintaining this same orientation show that sub-millimeter, discrete neuropathological features in marmoset brain consistently share size, shape, and location between histology and ex vivo MRI, which facilitates comparison with serial imaging acquired in vivo. Existing methods use computational approaches sensitive to data input in order to warp histologic images to match large-scale features on MRI, but the new method requires no warping of images, due to a preregistration accomplished in the technique, and is insensitive to data formatting and artifacts in both MRI and histology. The simple method of using 3D-printed brain holders to match brain orientation during pathologic sectioning and MRI acquisition enables rapid and precise comparison of small features seen on MRI to their underlying histology.


Characterization and anticancer effects of extracellular polysaccharide from DHA-producing microalga Crypthecodinium sp. SUN.
The Characterization and anticancer effects of extracellular polysaccharide (EPS) from DHA-producing microalga Crypthecodinium sp. SUN were studied in the present research. Results showed that EPS from C. sp. SUN have a molecular weight of 1.118 × 106 g/mol. EPS significantly inhibited the proliferation and migration of LA795 lung adenocarcinoma cells, and the apoptosis rate decreased in a concentration-dependent manner, reached 52 % at 15 mg/mL. C. sp. SUN EPS also significantly decreased reactive oxygen species (ROS) level by over 50 %, superoxide dismutase (SOD) activity by 76 %, and catalase (CAT) activity by 34 % at 10 mg/mL, indicating that EPS may inhibit tumor cell growth instead of killing tumor cells. Additionally, C. sp. SUN EPS suppressed cell proliferation by downregulating the expression of adhesion proteins and cyclin D1 in LA795 cells. In vivo experiments demonstrated that C. sp. SUN EPS inhibited the growth of lung adenocarcinoma tumors without affecting the normal body weight of nude mice. Collectively, the present study showed that C. sp. SUN EPS could be a potential substance for cancer treatment, which provided a research basis for future study on EPS and expanded the application of Crypthecodinium.


Thermographic assessment of impaired sympathetic function in peripheral nerve injuries.
Twenty-three patients with focal, strictly unilateral lesions of the peripheral nervous system were examined by infrared-thermography. The Minor sweat test was used to determine if sympathetic outflow was disturbed. In fifteen patients without a concomitant sympathetic lesion (controls) thermosymmetry was not disturbed. Eight patients had evidence of abnormal sweat secretion. In these patients thermoregulation was severely disturbed. During the first 5 to 8 months, affected skin areas were hyperthermic, whereas later only hypothermia was observed. Cold stimuli increased temperature differences in patients with disturbed sympathetic function, but not in controls. Thermography is a reliable, non-invasive technique to detect a lesion of sympathetic outflow and permits an estimation of the time-course of the lesion.


[ISGF3, a critical factor of the IFN-alpha pathway in the antiviral action of HBV].
To study the mechanism of signal transduction in anti-HBV action of IFN-alpha. The HBV DNA in HepG 2.2.15 cell line supernatant with/without IFNalpha-2b were monitored by fluorescence real-time quantitive PCR. STAT1, STAT2, ISGF3-gamma, PKR, 2'5'-OAS mRNA levels from HepG 2 and HepG 2.2.15 cell lines that were treated with/without IFNalpha-2b at different times were detected by semi-quantitive RT-PCR. And the ISGF3-gamma protein was detected by Western blot. Then, these items were detected again after inhibiting the JAK-STAT pathway with genistein. The HBV DNA in 2215 supernatant that were treated with IFNalpha-2b for 8 hours decreased 0.72 log 10 copies/ml. But the basal levels of DNA in cells pretreated with genistein? followed by IFNalpha-2b did not decrease. The STAT1, STAT2, ISGF3-gamma, 2'5'-OAS, PKR mRNA levels were upregulated by IFNalpha-2b. The same phenomena were observed with STAT1, STAT2, ISGF3-gamma mRNA when pretreated with genistein then treated with IFNalpha-2b, but the levels of 2'5'-OAS, PKR mRNA were decreased in this situation. The expression of the protein of ISGF3-gamma was also augmented by IFNalpha-2b, and was blocked by genistein. The JAK-STAT pathway seems to be a critical pathway in IFNalpha-2b action against HBV? and ISGF3 is most probably a key factor of the route.


Genetic modification strategies for enhancing plant resilience to abiotic stresses in the context of climate change.
Enhancing the resilience of plants to abiotic stresses, such as drought, salinity, heat, and cold, is crucial for ensuring global food security challenge in the context of climate change. The adverse effects of climate change, characterized by rising temperatures, shifting rainfall patterns, and increased frequency of extreme weather events, pose significant threats to agricultural systems worldwide. Genetic modification strategies offer promising approaches to develop crops with improved abiotic stress tolerance. This review article provides a comprehensive overview of various genetic modification techniques employed to enhance plant resilience. These strategies include the introduction of stress-responsive genes, transcription factors, and regulatory elements to enhance stress signaling pathways. Additionally, the manipulation of hormone signaling pathways, osmoprotectant accumulation, and antioxidant defense mechanisms is discussed. The use of genome editing tools, such as CRISPR-Cas9, for precise modification of target genes related to stress tolerance is also explored. Furthermore, the challenges and future prospects of genetic modification for abiotic stress tolerance are highlighted. Understanding and harnessing the potential of genetic modification strategies can contribute to the development of resilient crop varieties capable of withstanding adverse environmental conditions caused by climate change, thereby ensuring sustainable agricultural productivity and food security.


Certified reference materials for the determination of mycotoxins.
A continuing fundamental trend in the area of quality assurance in analytical laboratories are measures that enable trace-ability of analytical results to internationally renowned units, like e.g. the mole. Especially certified reference materials (CRMs) are important tools for achieving this goal. What are CRMs? Why do we need them in the field of mycotoxin analysis? Which kind of CRMs are currently available for the determination of mycotoxins and how should they be used? These are questions, which are increasingly being asked. This article attempts to answer these questions.


Respiration and substrate transport rates as well as reactive oxygen species production distinguish mitochondria from brain and liver.
Aberrant mitochondrial function, including excessive reactive oxygen species (ROS) production, has been implicated in the pathogenesis of human diseases. The use of mitochondrial inhibitors to ascertain the sites in the electron transport chain (ETC) resulting in altered ROS production can be an important tool. However, the response of mouse mitochondria to ETC inhibitors has not been thoroughly assessed. Here we set out to characterize the differences in phenotypic response to ETC inhibitors between the more energetically demanding brain mitochondria and less energetically demanding liver mitochondria in commonly utilized C57BL/6J mice. We show that in contrast to brain mitochondria, inhibiting distally within complex I or within complex III does not increase liver mitochondrial ROS production supported by complex I substrates, and liver mitochondrial ROS production supported by complex II substrates occurred primarily independent of membrane potential. Complex I, II, and III enzymatic activities and membrane potential were equivalent between liver and brain and responded to ETC. inhibitors similarly. Brain mitochondria exhibited an approximately two-fold increase in complex I and II supported respiration compared with liver mitochondria while exhibiting similar responses to inhibitors. Elevated NADH transport and heightened complex II-III coupled activity accounted for increased complex I and II supported respiration, respectively in brain mitochondria. We conclude that important mechanistic differences exist between mouse liver and brain mitochondria and that mouse mitochondria exhibit phenotypic differences compared with mitochondria from other species.


Pulmonary cement embolism complicating percutaneous kyphoplasty: A case report.
Vertebral cement augmentation procedures, as kyphoplasty (KP) or percutaneous vertebroplasty (PVP), are commonly used for the management of pain of the vertebral column usually due to fractures related to traumatic injury, osteoporosis or metastatic lesion. It is a useful and safe technique with few complications. Among them, symptomatic pulmonary cement embolism (PCE) can happened, even rarely described in the literature, leading to severe cardio-respiratory manifestations depending on the location and size of the cement emboli. A 55 yo woman presented with atypical chest pain and presyncope three weeks after a motor vehicle accident resulting in an L1 compression fracture treated with kyphoplasty. She was hemodynamically stable. Blood tests showed D-Dimer 0.29 μg/mL, troponin <5ng/mL, Brain Natriuretic Protein 14 ng/mL and a PaO2 of 99 mmHg on arterial blood gas analysis. A computed tomography scan of chest showed linear hyperdense foreign bodies in two segmental pulmonary arteries at the level of middle lobe and right lower lobe compatible with pulmonary cement embolism. Anticoagulation with rivaroxaban for sixth months resulted in resolution of symptoms. In this case, the pulmonary cement embolism occurred after kyphoplasty, which is associated with less risk of PCE than vertebroplasty. The procedure was done under biplanar fluoroscopy and no leakage of cement was noted, which would raise suspicion for CPE. Repeat imagine after this procedure is not routinely done. This case demonstrates that systematic imaging post procedure should be considered.An anticoagulation with rivaroxaban seems to be effective in our patient.


Smart vaccine delivery based on microneedle arrays decorated with ultra-pH-responsive copolymers for cancer immunotherapy.
Despite the tremendous potential of DNA-based cancer vaccines, their efficacious delivery to antigen presenting cells to stimulate both humoral and cellular response remains a major challenge. Although electroporation-based transfection has improved performance, an optimal strategy for safe and pain-free vaccination technique remains elusive. Herein, we report a smart DNA vaccine delivery system in which nanoengineered DNA vaccine was laden on microneedles (MNs) assembled with layer-by-layer coating of ultra-pH-responsive OSM-(PEG-PAEU) and immunostimulatory adjuvant poly(I:C), a synthetic double stranded RNA. Transcutaneous application of MN patches onto the mice skin perforate the stratum corneum with minimal cell damage; subsequent disassembly at the immune-cell-rich epidermis/dermis allows the release of adjuvants and DNA vaccines, owing to the ultra-sharp pH-responsive nature of OSM-(PEG-PAEU). The released adjuvant and DNA vaccine can enhance dendritic cell maturation and induce type I interferons, and thereby produce antigen-specific antibody that can achieve the antibody-dependent cell-mediated cytotoxicity (ADCC) and CD8+ T cell to kill cancer cells. Strikingly, transcutaneous application of smart vaccine formulation in mice elicited 3-fold greater frequencies of Anti-OVA IgG1 serum antibody and 3-fold excess of cytotoxic CD8+ T cell than soluble DNA vaccine formulation. As a consequence, the formulation rejected the murine B16/OVA melanoma tumors in C57BL/6 mice through the synergistic activation of antigen-specific ADCC and cytotoxic CD8+ T cells. The maneuvered use of vaccine and adjuvant poly(I:C) in MNs induces humoral and cellular immunity, which provides a promising vaccine technology that shows improved efficacy, compliance, and safety.


Pan-cancer single-cell analysis reveals the heterogeneity and plasticity of cancer-associated fibroblasts in the tumor microenvironment.
Cancer-associated fibroblasts (CAFs) are the predominant components of the tumor microenvironment (TME) and influence cancer hallmarks, but without systematic investigation on their ubiquitous characteristics across different cancer types. Here, we perform pan-cancer analysis on 226 samples across 10 solid cancer types to profile the TME at single-cell resolution, illustrating the commonalities/plasticity of heterogenous CAFs. Activation trajectory of the major CAF types is divided into three states, exhibiting distinct interactions with other cell components, and relating to prognosis of immunotherapy. Moreover, minor CAF components represent the alternative origin from other TME components (e.g., endothelia and macrophages). Particularly, the ubiquitous presentation of endothelial-to-mesenchymal transition CAF, which may interact with proximal SPP1+ tumor-associated macrophages, is implicated in endothelial-to-mesenchymal transition and survival stratifications. Our study comprehensively profiles the shared characteristics and dynamics of CAFs, and highlight their heterogeneity and plasticity across different cancer types. Browser of integrated pan-cancer single-cell information is available at https://gist-fgl.github.io/sc-caf-atlas/ .


Functional improvement in chronic stroke patients when following a supervised home-based computerized cognitive training.
Computerized Cognitive Training (CCT) is an effective treatment for cognitive impairment in the post-acute stage of stroke. However, it is still not clear if it is suitable for chronic stage. To explore if patients with cognitive deficit following stroke may benefit from CCT. Thirty patients post-stroke between 24 and 62 years old were randomized into two groups (A and B) to receive two different types of CCT. All patients were tested with a neuropsychological battery and functional questionnaires, before and after each CCT and also 6 months after the end of the study. In phase I, Group A received a customized CCT and Group B received a non-customized CCT, over 6 weeks. Three months after, each group received the other intervention (phase II). After phase I, between-group analyses revealed that Group A showed a relative decrease in subjective complaints. In contrast, Group B showed improvement in performance-based measures. After phase II, the decrease in subjective complaints continued in Group A, and both groups showed improvement in performance-based measures. Patients with chronic stroke improved cognitive functioning after performing supervised home-based multi-domain computerized cognitive training.


[Levamisol therapy in chronic prostatistis and epididymitis].
Eight patients suffering from chronic epididymitis, prostatis and urethritis were treated with levamisole. A definite and lasting improvement was seen six of the eight patients.


Immunogenicity of Mix-and-Match CoronaVac/BNT162b2 Regimen versus Homologous CoronaVac/CoronaVac Vaccination: A Single-Blinded, Randomized, Parallel Group Superiority Trial.
(1) Background: This study aimed to compare the immunogenicity of the mix-and-match CoronaVac/BNT162b2 vaccination to the homologous CoronaVac/CoronaVac regimen. (2) Methods: We conducted a simple-blinded randomized superiority trial to measure SARS-CoV-2 neutralization antibodies and anti-spike receptor binding domain (RBD) IgG concentrations in blood samples of participants who had received the first dose of CoronaVac vaccine followed by a dose of BNT162b2 or CoronaVac vaccine. The primary endpoint for immunogenicity was the serum-neutralizing antibody level with a percentage of inhibition at 90% at 21-35 days after the boost. A difference of 25% between groups was considered clinically relevant. (3) Results: Among the 240 eligible participants, the primary endpoint data were available for 100 participants randomly allocated to the mix-and-match group versus 99 participants randomly allocated to the homologous dose group. The mix-and-match regimen elicited significantly higher levels of neutralizing antibodies (median level of 96%, interquartile range (IQR) (95-97) versus median level of 94%, IQR (81-96) and anti-spike IgG antibodies (median level of 13,460, IQR (2557-29,930) versus median level of 1190, IQR (347-4964) compared to the homologous group. Accordingly, the percentage of subjects with a percentage of neutralizing antibodies > 90% was significantly higher in the mix-and-match group (90.0%) versus the homologous (60.6%). Interestingly, no severe events were reported within 30 days after the second dose of vaccination in both groups. (4) Conclusions: Our data showed the superiority of the mix-and-match CoronaVac/BNT162b2 vaccination compared to the CoronaVac/CoronaVac regimen in terms of immunogenicity, thus constituting a proof-of-concept study supporting the use of inactivated vaccines in a mix-and-match strategy while ensuring good immunogenicity and safety.


Preprocedural Thrombolysis in Myocardial Infarction (TIMI) flow significantly affects the extent of ST-segment resolution and myocardial blush in patients with acute anterior myocardial infarction treated by primary angioplasty.
Although the benefits of primary angioplasty for ST-segment elevation myocardial infarction (STEMI) have been demonstrated, the prognostic role of early recanalization in these patients has yet to be investigated. The aim of the study was to evaluate the impact of preprocedural Thrombolysis in Myocardial Infarction (TIMI) flow on the extent of myocardial reperfusion in patients with anterior STEMI treated with primary angioplasty. Our population consisted of 754 consecutive patients with anterior STEMI treated by primary angioplasty from April 1997 to October 2001. All angiographic, clinical, and follow-up data were prospectively collected. Preprocedural TIMI flow was related to postprocedural TIMI 3 flow (P < .0001), ST-segment resolution (P = .009), myocardial blush grade (P < .0001), enzymatic infarct size (P < .0001), and predischarge ejection fraction (P < .0001), even in the analysis restricted to patients with postprocedural TIMI 3 flow. These data explain the observed significant impact of preprocedural TIMI flow on 1-year mortality. This study shows that in patients with anterior STEMI, poor preprocedural TIMI flow is associated with impaired perfusion, larger infarct size, and 1-year mortality. This study suggests that all efforts should be made to obtain optimal restoration of antegrade flow as early as possible before angioplasty. Further studies are needed to investigate the impact of early adjunctive pharmacological therapy on preprocedural TIMI flow and mortality in these high-risk patients.


Manganese-enhanced MRI of the optic visual pathway and optic nerve injury in adult rats.
To evaluate manganese (Mn2+)-enhanced MRI in a longitudinal study of normal and injured rat visual projections. MRI was performed 24 hours after unilateral intravitreal injection of MnCl2 (150 nmol) into adult Fischer rats that were divided into four groups: 1) controls (N = 5), 2) dose-response (N = 10, 0.2-200 nmol), 3) time-response with repeated MRI during 24-168 hours post injection (N = 4), and 4) optic nerve crush (ONC) immediately preceding the MnCl2 injection (N = 7). Control and ONC animals were reinjected with MnCl2 20 days after the first injection, and MRI was performed 24 hours later. In the control group, the optic projection was visualized from the retina to the superior colliculus, with indications of transsynaptic transport to the cortex. There was a semilogarithmic relationship between the Mn2+ dose and Mn2+ enhancement from 4 to 200 nmol, and the enhancement decayed gradually to 0 by 168 hours. No Mn2+-enhanced signal was detected distal to the ON crush site. In the control group, similar enhancement was obtained after the first and second MnCl2 injections, while in the ONC group the enhancement proximal to the crush site was reduced 20 days post lesion (20 dpl). Mn2+-enhanced MRI is a viable method for temporospatial visualization of normal and injured ON in the adult rat. The observed reduction in the Mn2+ signal proximal to the ONC is probably a result of retrograde damage to the retinal ganglion cells, and not of Mn2+ toxicity.


Ethics instruction in an issues-oriented course on public health genetics.
Public health ethics and genetics in public health have in the last decade emerged as fields in their own right. This article describes the ethics component of an "Issues in Public Health Genetics" course uniting the fields of genetics, ethics, and public health. As the third member of the University of Michigan Interdepartmental Concentration in Genetics, the course emphasizes the translation of ethical-social issues into substantive policies. A central tension repeated throughout the course exists between respect for individuals' libertarian interests and the welfare of the entire population. Students become familiar with the process of resolving conflicting values through interactive exercises addressing the health needs of diverse communities. Key lessons in course design are described.


Pregnenolone metabolism in Y-1 mouse adrenal cells: HPLC analysis and identification of metabolites by gas chromatography-mass spectrometry.
The Y-1 mouse adrenocortical tumor cells secrete into the culture medium some metabolites of pregnenolone. The long term basic steroid secretion was studied for 24 h by high performance liquid chromatography (HPLC). After 6 h incubation, only one chromatographic peak was shown to be quantitatively important. When the cells were incubated with tritium labeled pregnenolone and carrier, the analysis of radioactivity of each HPLC fraction indicated a rapid incorporation of the exogenous precursor and successive secretion/reabsorption kinetics of the metabolites. The final product, accumulated in the medium, was characterized by gas chromatography-mass spectrometry as 11 beta,20 alpha-dihydroxy-4-pregnen-3-one.


MicroRNA 155 modulates megakaryopoiesis at progenitor and precursor level by targeting Ets-1 and Meis1 transcription factors.
MicroRNAs (miRNAs) control basic biological functions and are emerging as key regulators of haematopoiesis. This study focused on the functional role of MIRN155 on megakaryocytic (MK) differentiation of human cord blood CD34+ haematopoietic progenitor cells (HPCs). MIRN155, abundantly expressed in early HPCs, decreases sharply during MK differentiation. Functional studies showed that enforced expression of MIRN155 impairs proliferation and differentiation of MK cells. Furthermore, HPCs transfected with MIRN155 showed a significant reduction of their MK clonogenic capacity, suggesting that down-modulation of this miRNA favours MK progenitor differentiation. Consistent with this observation, MIRN155 downregulates, by directly binding to their 3'-UTR, the expression of Ets-1 and Meis1, two transcription factors with well-known functions in MK cells. These results show that the decline of MIRN155 is required for MK proliferation and differentiation at progenitors and precursors level and indicate that sustained expression of MIRN155 inhibits megakaryopoiesis.


Proteomic biomarkers in lung cancer.
The correct understanding of tumour development relies on the comprehensive study of proteins. They are the main orchestrators of vital processes, such as signalling pathways, which drive the carcinogenic process. Proteomic technologies can be applied to cancer research to detect differential protein expression and to assess different responses to treatment. Lung cancer is the number one cause of cancer-related death in the world. Mostly diagnosed at late stages of the disease, lung cancer has one of the lowest 5-year survival rates at 15 %. The use of different proteomic techniques such as two-dimensional gel electrophoresis (2D-PAGE), isotope labelling (ICAT, SILAC, iTRAQ) and mass spectrometry may yield new knowledge on the underlying biology of lung cancer and also allow the development of new early detection tests and the identification of changes in the cancer protein network that are associated with prognosis and drug resistance.


Image: Extensive Pneumocephalus.
A 53-year-old man was admitted to our hospital following a traffic accident. He had been riding a bicycle and was knocked down by a motorcycle, injuring the right side of his forehead. Upon arrival at our department, he was conscious and we found no cerebrospinal fluid rhinorrhea or otorrhea. His Glasgow Coma Scale score was 15. Neurologic examination was normal, except for reduced vision in his right eye. Head computed tomography showed extensive pneumocephalus involving the cisterna ambiens, prepontine and suprasellar cisterns, and temporal, frontal, parietal, and occipital lobes. A right frontal skull fracture was evident. The patient was treated conservatively, with bed rest with the head of the bed elevated at 30 degrees. The patient was instructed to avoid any movements that might increase intracranial pressure, and he was placed on conventional concentration oxygen therapy of about 40%. Repeat computed tomography 9 hours later showed partial absorption of the pneumocephalus, which was almost completely absorbed 4 days later. He recovered well and was discharged after 7 days. At the 1-month follow-up, the patient was well, although the reduced vision in his right eye persisted.


Structural in silico dissection of the collagen V interactome to identify genotype-phenotype correlations in classic Ehlers-Danlos Syndrome (EDS).
Collagen V mutations are associated with Elhers-Danlos syndrome (EDS), a group of heritable collagenopathies. Collagen V structure is not available and the disease-causing mechanism is unclear. To address this issue, we manually curated missense mutations suspected to promote classic type EDS (cEDS) insurgence from the literature and performed a genotype-phenotype correlation study. Further, we generated a homology model of the collagen V triple helix to evaluate the pathogenic effects. The resulting structure was used to map known protein-protein interactions enriched with in silico predictions. An interaction network model for collagen V was created. We found that cEDS heterogeneous manifestations may be explained by the involvement in two different extracellular matrix pathways, related to cell adhesion and tissue repair or cell differentiation, growth and apoptosis.


Predictability of what or where reduces brain activity, but a bottleneck occurs when both are predictable.
Detecting regularities in the sensory environment licenses predictions that enable adaptive behaviour. However, it is unclear whether predictions about object category, location, or both dimensions are mediated by overlapping systems, and relatedly, whether constructing predictions about both category and location is associated with processing bottlenecks. To examine this issue, in an fMRI study, we presented participants with image-series in which non-deterministic transition probabilities enabled predictions about either the location of the next image, its semantic category, both dimensions, or neither (the latter forming a "no-regularity" random baseline condition). Speaking to a common system, all three predictable conditions resulted in reduced BOLD activity in four clusters: left rostral anterior cingulate cortex; bilateral putamen, caudate and thalamus; right precentral gyrus, and left visual cortex. Pointing to a processing bottleneck, in some regions, a significant interaction between the two factors was found whereby category-predictable series were associated with lower activity - but only when location regularity was absent. Finally, category regularity decreased activation in areas of the ventral visual stream and semantic areas of lateral temporal cortex, and location regularity decreased activation in a dorsal fronto-parietal cluster, long implicated in the endogenous control of spatial attention. Our findings confirm and expand a role for dACC/dmPFC and striatum in monitoring or responding to uncertainty in the environment and point to a limited capacity bottleneck when multiple predictions are concurrently licensed.


Reproductive system and colonizing strategy of two species of Carduus (compositae).
Carduus pycnocephalus and C. tenuiflorus are two important weed species in mediterranean-type ecosystems. They were studied in their native habitat in southern France from April to December 1981.These species both produce two types of seed (achenes) in each capitulum. Those in the center are not dormant and are easily dispersed, while the outer ones can be dormant and have no apparent means of dispersal. Our observations on pollen transfer and self fertilization show that the dispersable, non dormant seeds have a low probability of inbreeding, while the potentially dormant, non dispersed seeds have a high probability of inbreeding. The significance of this dimorphism is discussed.