Exploring the potential of salivary and blood immune biomarkers to elucidate physical frailty in institutionalized older women.
Identification of older populations at increased risk of physical frailty using biochemical approaches could improve screening accuracy. The aim of this study was to study the relationship between immune markers and independent components of physical frailty in institutionalized older women. A sample of 358 institutionalized-dwelling women, aged 75 years and older, were assessed for biosocial factors and general health status, pro and anti-inflammatory cytokines, sex steroid hormones, salivary anti-microbial proteins, blood cells counts and the five Fried's physical frailty components that allowed for classification of the sample into frail, prefrailty and not-frail subgroups. Results showed that cytokines IL-6, IL-10, IL-1β, TNF-α, and the TNF-α/IL-10 ratio, mean corpuscular haemoglobin, salivary cortisol and α-amylase were all associated with frailty. Weakness and Exhaustion were the frailty components that were most strongly associated with these biomarkers. Salivary α-amylase was the biomarker that best explained frailty, as it was associated with all five components of physical frailty, and could be used as a potential screening tool. Future research needs to investigate the causal-effect association between salivary innate immune makers, susceptibility to infection and frailty.


Expression of phosphorylated eIF4E-binding protein 1, but not of eIF4E itself, predicts survival in male breast cancer.
Male breast cancer is rare and treatment is based on data from females. High expression/activity of eukaryotic initiation factor 4E (eIF4E) denotes a poor prognosis in female breast cancer, and the eIF4E pathway has been targeted therapeutically. Eukaryotic initiation factor 4E activity in female breast cancer is deregulated by eIF4E overexpression and by phosphorylation of its binding protein, 4E-BP1, which relieves inhibitory association between eIF4E and 4E-BP1. The relevance of the eIF4E pathway in male breast cancer is unknown. We have assessed expression levels of eIF4E, 4E-BP1, 4E-BP2 and phosphorylated 4E-BP1 (p4E-BP1) using immunohistochemistry in a large cohort of male breast cancers (n=337) and have examined correlations with prognostic factors and survival. Neither eIF4E expression nor estimated eIF4E activity were associated with prognosis. However, a highly significant correlation was found between p4E-BP1 expression and disease-free survival (DFS), linking any detectable p4E-BP1 with poor survival (univariate log rank P=0.001; multivariate HR 8.8, P=0.0001). Our data provide no support for direct therapeutic targeting of eIF4E in male breast cancer, unlike in females. However, as p4E-BP1 gives powerful prognostic insights that are unrelated to eIF4E function, p4E-BP1 may identify male breast cancers potentially suitable for therapies directed at the upstream kinase, mTOR.


The Xmas-2 Protein of Drosophila melanogaster Undergoes Cleavage into Two Fragments.
The TREX-2 complex integrates several stages of gene expression, such as transcriptional activation and mRNA export. In D. melanogaster, TREX-2 consists of four major proteins: Xmas-2, ENY2, PCID2, and Sem1p. The Xmas-2 protein is the core subunit of the complex, with which other TREX-2 subunits interact. Xmas-2 homologues were found in all higher eukaryotes. Previously, it was shown that the human Xmas-2 homologue, GANP protein, can undergo cleavage into two parts, probably during apoptosis. We showed that the Xmas-2 protein of D. melanogaster can also split into two fragments. The resulting fragments of the protein correspond to the two large Xmas-2 domains. Protein splitting is observed both in vivo and in vitro. However, Xmas-2 cleavage in D. melanogaster is observed under normal conditions and is probably a part of the mechanism of transcription and mRNA export regulation in D. melanogaster.


Treatment with an apolipoprotein A-1 mimetic peptide in combination with pravastatin inhibits collagen-induced arthritis.
To evaluate the therapeutic potential of an apolipoprotein A-1 (apoA-1) mimetic peptide, D-4F, in combination with pravastatin in collagen-induced arthritis (CIA), syngeneic Louvain rats were immunized with type II collagen and randomized to vehicle control, D-4F monotherapy, pravastatin monotherapy, or D-4F + pravastatin combination therapy. Clinical arthritis activity was evaluated and radiographs, type II collagen antibody titers, cytokine/chemokine levels, and HDL function analysis were obtained. There was significant reduction in clinical severity scores in the high and medium dose D-4F + pravastatin groups compared to controls (p< or =0.0001). Reduction in erosive disease occurred in the medium/high dose combination groups compared to non-combination groups (p< or =0.01). Favorable changes in cytokines/chemokines were noted with treatment, and response to combination D-4F/pravastatin therapy was associated with improvement in HDL's anti-inflammatory properties. Combination D-4F/pravastatin significantly reduced clinical disease activity in CIA, and may have dual therapeutic potential in other autoimmune diseases with increased cardiovascular morbidity and mortality.


Inactivation of a slow Ca2+ current in CA1 neurones of the adult rat hippocampal slice.
CA1 neurones of the adult rat hippocampal slice preparation were voltage clamped at or near -40 mV membrane potential using a single electrode clamp method. Depolarizing voltage commands from a holding potential of -40 mV elicited voltage-dependent inward Ca2+ currents comprising a fast and a slow component. The latter one was investigated for its susceptibility to inactivation, which was maximally expressed at around 0 mV membrane potential. When extracellular Ca2+ was replaced by Ba2+, inward currents became much larger and were followed by long tail currents. Similar data were observed in neurones injected with the Ca2+ chelator BAPTA. It is suggested that inactivation of the slow Ca2+ current depends at least partly on the levels of intracellular free Ca2+ in hippocampal neurones.


Image analysis of pubic bone for sex determination in a computed tomography sample.
Radiology has demonstrated great utility for sex determination, but most studies are based in metrical and morphological methods in order to perform an identification profile. It is presented an easy image analysis-based method aimed to demonstrate the relationship between the bony tissue ultrastructure and sex by using several grey level histogram variables obtained from computed tomography images using sagittal sections of the pubic symphysis surface and the body of the pubis. The CT sample consisted of 169 hospital DICOM archives of known sex and age. The multivariate binary logistic regression models calculated showed reliabilities for sex determination between 87.4 and 95.8 %, with a high intra and inter-observer agreement. The suggested method is not only considered useful to perform an identification profile during Virtopsy, but even for application in further studies in order to attach a quantitative correlation for tissue ultrastructure characteristics without complex and expensive methods beyond image analysis.


Evidence supporting the independent inheritance of primary affective disorders and primary alcoholism in the families of bipolar patients.
This study explored the nature of the association between bipolar disorder and alcoholism. The authors studied 814 first-degree relatives of 121 bipolar patients, divided on the basis of response to lithium prophylaxis. Logistic regression analysis was used to analyze the contribution of demographic, familial and clinical variables to the risk of primary alcoholism in the relatives. The risk of primary alcoholism in relatives was not related to the degree of affective loading in the family or to the proband's lithium response. This study does not support a shared genetic liability between bipolar disorder and alcoholism. This study lacked a control group, but the analysis accounted for this. These disorders are not alternative forms of the same illness.


Prenatal O3 exposure increases the severity of OVA-induced asthma in offspring.
Accumulating epidemiological studies showed that prenatal and early life exposure to ambient air pollution was important contributor to the development of childhood asthma. However, the effects and mechanisms of prenatal exposure to ozone (O3), a type of ambient air pollution, on the progression of asthma in offspring remain unclear. This study aimed to determine the effects and mechanism of asthma in offspring after prenatal O3 exposure. Pregnant BALB/c mice were exposed to O3 or air on gestational days (GDs) 13-18. Their offspring were sensitized and challenged to ovalbumin (OVA) to establish asthma model, and the asthma features were evaluated. The splenic natural killer (NK) cells in the offspring were measured to explore the mechanism on the effects of asthma in the offspring. The responses of the pregnant mice and dams after O3 exposure were evaluated. Airway inflammation, mucus secretion, OVA-specific immunoglobulin (Ig) E, T helper (Th) 2-skewed response, the frequency of CD3ε-CD49b+ splenic NK cells, the expression of tumor necrosis factor (TNF)-α, and IL (interleukin)-17 were significantly exacerbated in the OVA-induced asthma offspring after prenatal O3 exposure. In addition, airway inflammation, a lower number of CD3ε-CD49b+ splenic NK cells, and systemic oxidative stress were caused at the end of pregnancy after O3 exposure, which did not recover at the end of lactation for the first two responses. Prenatal O3 exposure increased the severity of OVA-induced asthma in the offspring, which might be directly induced by CD3ε-CD49b+ splenic NK cells in the offspring and indirectly related to the damaged maternal immune system.


Utilizing preoperative arterial embolization to minimize blood loss at time of vulvar sarcoma resection: A case report.
Selective arterial embolization has long been utilized in the field of obstetrics and gynecology as both a prophylactic and therapeutic measure. We present a case of a highly vascularized, poorly differentiated, radiation-related vulvar sarcoma that was resected immediately following selective arterial embolization. Arterial embolization in patients with gynecologic malignancies has been shown to compromise blood supply to the tumor and decrease the risk of hemorrhage and related surgical complications. In this case, the use of embolization prior to surgery appeared to result in a less morbid procedure for the patient with decreased blood loss and improved quality of life thereafter. Based on the presented case, we believe that arterial embolization can be considered a safe preoperative intervention to decrease surgical risk, particularly hemorrhage, at time of resection of highly vascularized vulvar tumors.


The insulin-dependent diabetes mellitus-associated ICA 105 autoantigen in stiff-man syndrome patients.
The pathogenetic role of anti-glutamic acid decarboxylase (GAD) antibodies found in up to 60% of patients with stiff-man syndrome (SMS) is still controversial. GAD, in fact, is also one of the major target antigen of insulin-dependent diabetes mellitus (IDDM), a disease affecting one third of anti-GAD antibody-positive patients with SMS. To better define the role of autoimmunity in SMS we looked for molecular and immunological evidence of an autoimmune recognition of a second IDDM-associated autoantigen, the pancreatic 37/40 kDa IDDM-autoantigen, whose gene called ICA 105 has been recently cloned. By Northern blot analysis we found that tissue distribution of human ICA 105 is restricted to pancreas and brain and within the central nervous system (CNS) its distribution is similar to GAD. We also measured anti-ICA 105 antibodies in 11 SMS patients and 56 control patients with other neurological diseases (OND). Anti-ICA 105 antibodies were found in 4/11 (36%) patients with SMS (a frequency similar to that of anti-GAD-antibodies in our SMS population) but in only 2/56 (3%) patients with OND (P < 0.001). Anti-ICA 105 and anti-GAD antibodies were associated in 3/4 (75%) patients with SMS but in none of the 2 anti-ICA 105 antibody-positive OND patients. Among anti-ICA 105 antibody-positive patients with SMS, only 1 suffered also from IDDM. In contrast, the only 2 anti-ICA 105 antibody-positive with OND had IDDM. Our results indicate that ICA 105 represents another putative neuroendocrine autoantigen in SMS. The presence of circulating anti-GAD and/or anti-ICA 105 antibodies might help the diagnosis of SMS. The absence, however, of antibodies recognising specific CNS autoantigens (e.g. GAD, ICA 105) does not rule out SMS.


Early destruction of cockroach respiratory system and heart by emerald jewel wasp larvae.
In the 1880s, Henri Fabre was captivated by the "special art of eating", whereby a parasitoid wasp larva fed selectively on host internal organs, avoiding the heart (dorsal vessel) and tracheal system (respiratory system) to preserve life. In Fabre's words: "The ruling feature in this scientific method of eating, which proceeds from parts less to the parts more necessary to preserve a remnant of life, is none the less obvious"1. Subsequent investigators have reported the same for many parasitoid wasps2,3, including for the emerald jewel wasp (Ampulex compressa)4. Here it is reported that larval jewel wasps destroy the dorsal vessel and tracheae (respiratory system) in the thorax of their cockroach host (Periplaneta americana) at their earliest opportunity. Moreover, the broken tracheae release air into the host, which the larval jewel wasp inspires. An increase in larval chewing rate, cotemporaneous with the sudden release of air from the host's broken tracheae, suggests the larva taps into the host respiratory system to support its metabolism while rapidly consuming the host. VIDEO ABSTRACT.


Exploring the fMRI based neural correlates of the dot probe task and its modulation by sex and body odor.
The dot probe task implicitly cues attention via emotional information, an effect which is especially pronounced for threat-related cues. However, several questions remain unexplored. The first one is whether chemosignals like the androgen-derivative androstadienone can influence such attentional biases. Second, few studies have addressed sex differences regarding attentional biases. Finally, the neural correlates of these potential behavioral effects based on functional magnetic resonance imaging (fMRI) are not known. In two experiments we aimed to answer these questions. A total of 159 healthy individuals (58 oral-contraceptive-users, 42 luteal women, 59 men) were tested. In experiment 1 (behavioral study) we examined attentional biases behaviorally, while in experiment 2 (fMRI study) the dot probe task was complemented by fMRI. Our results provide robust evidence that in healthy participants fearful but not angry or happy faces lead to a strong general attentional bias. Elucidating the neural basis of this effects points to an early processing advantage in bilateral thalamus for valid compared to invalid cued fear. However, this finding was limited to those participants with the strongest attentional biases and was not linked to behavioral measures. Furthermore, no consistent sex or group differences existed neither did the putative human chemosignal androstadienone reliably modulate attentional biases or change neural processing.


Effectiveness of an educational program to promote pesticide safety among pesticide handlers of South India.
Occupational poisoning with pesticides is common in developing countries because farmers are often under trained, illiterate and consider it impractical and expensive to use safety equipment, especially in tropical climates. Greater benefit of education programs on prevention can be obtained if initiated in areas having higher occurrence of poisoning. Hence, the present study evaluated occurrence of poisoning and effectiveness of educational interventions among pesticide handlers in areas having high occurrence of occupational poisoning. Two villages of Udupi district of South India were identified by spot mapping and targeted for a public education program on safe handling of pesticides, the impact of which was assessed using a knowledge attitude and practice (KAP) questionnaire. Education was provided using a structured individualized training program to 74 pesticide handlers. Three point KAP assessments were carried out at baseline, immediately after training and after 1 month of training. Nonparametric Kruskal-Wallis tests and Friedmann tests were used to compare scores at different time points and between groups. Occurrence of occupation related poisoning was 33% and common in three villages of the district. The average baseline KAP score of 30.88 +/- 10.33 improved after education significantly (P < 0.001) at first follow-up 45.03 +/- 9.16 and at second follow-up 42.9 +/- 9.54. A decline of score between the first and second follow-up may be attributed to decline in knowledge retention. Demographics like gender, literacy and presence of children affected KAP score and there was no influence of geography, age or frequency of pesticide use. Educational intervention among pesticide handlers improved the KAP score for safe pesticide handling. We recommend that continuous education and training programs for agricultural workers will promote awareness and minimize the hazards of occupational pesticide exposure.


Echocardiographic evaluation in asymptomatic relatives of patients with dilated cardiomyopathy reveals preclinical disease.
Idiopathic dilated cardiomyopathy is often familial, and apparently healthy relatives may have latent, early, or undiagnosed established disease. To determine the prevalence and natural history of asymptomatic cardiac abnormalities among sampled relatives of unselected patients referred for management of dilated cardio-myopathy. Prospective cohort study. 767 asymptomatic relatives of 189 consecutive unselected patients with dilated cardiomyopathy. Clinical evaluation, including history, physical examination, electrocardiography, and echocardiography, was performed. Participants were classified in accordance with published echocardiographic criteria. Sampled relatives who did not have evidence of dilated cardiomyopathy at the initial evaluation were followed for a median of 57 months (range, 1 to 133 months). Of the 767 patients evaluated, 592 (77.2%) were assessed as healthy, 35 (4.6% [95% CI, 3.7% to 7.6%]) had dilated cardiomyopathy, 119 (15.5% [CI, 12.5% to 18.8%]) had left ventricular enlargement without systolic dysfunction, and 21 (2.7% [CI, 1.9% to 4.9%]) had depressed fractional shortening without ventricular dilatation. At follow-up, progression to dilated cardiomyopathy occurred in 13 (10%) relatives with left ventricular enlargement or depressed fractional shortening versus 3 (1.3%) healthy relatives. In a multivariate model, only left ventricular enlargement or depressed fractional shortening independently predicted progression to dilated cardiomyopathy (hazard ratio, 10.0 [CI, 2.8 to 35.5]; P < 0.001). Because relatives had to be willing to participate and be available geographically, selection bias may have occurred. Treatable asymptomatic dilated cardiomyopathy was identified in 4.6% of asymptomatic relatives. In addition, left ventricular enlargement and depressed fractional shortening were common in asymptomatic relatives of patients with dilated cardiomyopathy and were associated with a statistically significant medium-term risk for disease progression. Evaluation of relatives of patients with cardiomyopathy is recommended.


Erectile dysfunction in patients with nonalcoholic fatty liver disease.
There is a lack of studies on erectile dysfunction (ED) in patients diagnosed with nonalcoholic fatty liver disease (NAFLD). The present study aimed to estimate the prevalence of ED in patients with NAFLD and to determine the independent predictors of ED in these patients. We conducted a prospective, hospital-based study of 192 consecutive male patients with NAFLD. All patients underwent clinical evaluation; abdominal ultrasonography; test for viral hepatitis markers; and estimation of liver chemistry panel, complete blood count, prothrombin time, serum lipids panel, serum testosterone, and fasting serum levels of glucose, insulin, and C-peptide. The mean age of the study population was 42.4±7.7years (79.1%≥40years). Of the 192 patients with NAFLD, 88 (45.8%) had ED, 28 (14.6%) had metabolic syndrome, 25 (13%) had type-2 diabetes mellitus (DM), and 131 (68.2%) had insulin resistance (IR). The mean level of serum testosterone was 3.17±2.94ng/mL, while the mean insulin resistance index was 2.9±1.7. Mild ED (38.6%) was the most frequent grade of ED. Age≥40years (odds ratio [OR] 6.4; 95% confidence interval [CI] 1.7-24.1; p- 0.006), IR (OR 5.9; 95% CI 1.7-20.6; p- 0.005), and low serum testosterone (OR 5.1; 95% CI 1.5-17.1; p- 0.009) were the predictors of ED. ED is a common disorder in male patients with NAFLD; both IR and low serum testosterone contribute to its development. Treatment of IR may carry a dual benefit of improving erectile function and decreasing the grade of hepatic steatosis.


Modeling windthrow effects on water runoff and hillslope stability in a mountain catchment affected by the VAIA storm.
Windthrows seriously affect forest landscapes, causing several issues in hydrological and geomorphological terms. In this regard, Airborne Laser Scanning (ALS) topographic data recently increased the opportunity to investigate in detail physical processes at the catchment scale. Moreover, topographically based hydrological and geomorphological models allow quantifying runoff alteration due to windthrows-driven land cover changes and detect the occurrence of land degradative processes at the sub-catchment scale. In this connection, accurate investigations about windthrows role in varying local runoff regimes over time are still obscure, as well as the possibility of predicting terrain instabilities due to windstorm occurrence. This research aims to investigate the interaction between windthrows, runoff alterations and hillslope failures affecting a landslide-prone mountain catchment (northern Italy). Hydrological HEC-HMS and geomorphological RESS models were applied. Windthrows' role in altering runoff regimes and hillslope stability was investigated starting from the elaboration of ALS-derived points clouds acquired before and after the occurrence of the Vaia storm. Digital Terrain Models (DTMs) were elaborated for the two scenarios to compare daily runoff variations and predict the activation of terrain instabilities by looking at land cover changes driven by the blowdown event at the sub-catchment detail. Results attested the key role of windstorms in altering local runoff, with a maximum relative runoff increment equal to 2.56 % and a maximum runoff difference equal to 3.12 mmh-1, as well as in encouraging the activation of the observed shallow landslide. The correlation between windthrows occurrence and runoff alterations was validated by performing regression analysis (R2 = 0.76), while the accuracy of instabilities predictions was tested through the Distance to Perfect Classification (D2PC) index and True Skill Statistic (TSS) score, respectively resulted equal to 0.076 and 0.898. This research represents a valid tool for investigating similar issues at a wider scale, also providing suggestions for promoting interventions in wind-disturbed forest areas.


[Severe poisoning with the organophosphorus insecticide thionazine--2 cases with the development of adult respiratory distress syndrome (ARDS)].
Among the insecticides, organophosphorous compounds are the agents most involved in deliberate and involuntary intoxications. The toxic effect is mainly due to the inhibition of cholinesterase at the cholinergic synapses. Two cases of severe involuntary intoxication are described which were characterized by the development of ARDS.


Effect of somatostatin on presynaptic and postsynaptic glutamate receptors and postsynaptic GABA receptors in the neurons of rat brain.
We studied the effect of somatostatin on presinaptic NMDA receptors and postsinaptic GABA, NMDA, and AMPA receptors in rat brain. It was shown that somatostatin inhibits NMDA-induced (45)Ca(2+) uptake into synaptosomes isolated from rat brain cortex (IC50=2.8×10(-11) M). Somatostatin potentiates AMPA receptors and inhibits hippocampal NMDA receptors in the entire range of examined concentrations (10(-14)-10(-7) M); it also potentiates or inhibits GABA receptor currents in a concentration-dependent manner. Our results suggest that somatostatin modulates the function of ionotropic glutamate and GABA receptors and is involved in cognitive and neurodegenerative processes in the mammalian brain.


Are Biomphalaria snails resistant to Schistosoma mansoni?
Among Biomphalaria glabrata/Schistosoma mansoni snail-trematode combinations, it appears that some parasites succeed whilst others fail to infect snails. Snails that become infected are termed susceptible hosts. Those which are not infected are traditionally determined as 'resistant'. Here the concept of B. glabrata resistance to S. mansoni is re-examined in the light of additional observations. It is suggested that, in B. glabrata/S. mansoni, compatibility is tested independently for each individual miracidium and host, and that the success or failure of an infection does not depend on the snail susceptibility/resistance status, but on the 'matched' or 'mismatched' status of the host and parasite phenotypes.


Evidence for multiple genes coding for the isozymes of hexokinase in the highly glycolytic AS-30D rat hepatoma.
We have compared Southern blots of rat hepatoma DNA probed with Types I, II and III hexokinase cDNAs isolated from normal rat tissues. Hybridization patterns show several fragments recognized by both the Type I and II clones while no resemblance is observed between the Type III probe and the other two isozymes. It therefore appears that the Type I-like and Type II-like hepatoma isozymes are coded for by similar yet separate genes, while a dissimilar third gene codes for the Type III-like isozyme. In addition, a loss of heterozygosity was detected at the Type III locus in the AS-30D hepatoma when compared to normal tissue. As only the Type II-like isozyme is highly expressed in highly glycolytic tumors, these data have implications for differential gene regulation between the tumor isozymes.


The energy budget of Porcellio spinicornis Say (Porcellionidae, Isopoda) under laboratory conditions.
The rate of energy flow in Porcellio spinicornis was investigated in a select group of a laboratory population. The consumption, assimilation and production were calculated to be 1 233, 1 002 and 304 cal/year, respectively, in the male, and 1 261, 1 198 and 429 cal/year, respectively, in the female. Ignoring loss due to mortality, and applying the population density of 2 295 m2 in the laboratory colony, the isopod population should consume 2 861.8 kcal m-2 year-1, resulting in an assimilation of 2 524.5 kcal m-2 year-1 and a total productivity of 841.1 kcal m-2 year-1.


(Queer) Family Values and "Reciprocal IVF": What Difference Does Sexual Identity Make?
In this paper I employ the case of "reciprocal IVF" (R-IVF)-in which a female-female couple uses in vitro fertilization to allow one woman to be the genetic mother and the other the gestational mother of their child(ren)-to illuminate the role sexual identity might productively play in bioethics. Bioethicists who have taken up this issue have largely focused on the moral permissibility and availability of the technology, and so defend R-IVF through analogy to commonly accepted different-sex uses of IVF. In this way, they position sexual identity as largely irrelevant to the primary bioethical questions raised by R-IVF. My approach diverges on these counts as I focus on the ethics of R-IVF through the lens of queer family values specifically. I ask what the practice of R-IVF as an option might mean for the queer community at large, and so view sexual identity as integral to a full bioethical evaluation of the practice.


Dental services during the COVID-19 pandemic: A tertiary care hospital experience.
The Aga Khan University is the largest tertiary care hospital in the biggest metropolis of Pakistan, in this paper six weeks of experience during the COVID-19 pandemic is shared, we will discuss the measures that were taken to mitigate the spread of COVID-19 amongst dental health care providers (DHCPS) and provide a neoteric workflow for the provision of safe dental care as dental services move towards normalcy. Furthermore topic such as fit testing, contact tracing, employee health and training are discussed and experience of such measures is shared. It is hoped that till the time new evidence based data is generated these communications may help spread awareness to help setup clinical protocols for other tertiary care settings.


Growth and analysis of crystal forms of toxic shock syndrome toxin 1.
Native toxic shock syndrome toxin 1 (TSST-1) purified from Staphylococcus aureus has been crystallized in four different forms. The highest resolution data (2.05 A) was collected from orthorhombic crystals belonging to the space group C222(1). The unit cell dimensions are a = 108.7 A, b = 177.5 A, c = 97.6 A. Rotation function analysis of this form indicates that there is trimer of toxin molecules in the asymmetric unit with a local 3-fold axis parallel to the crystallographic c axis. Crystals of a double mutant of TSST-1 have been grown which has a single molecule in the asymmetric unit and diffract to 1.9 A. The space group is P2(1) with unit cell parameters of a = 44.4 A, b = 34.0 A, c = 55.2 A, beta = 93.0 degrees.


Analysis of the molecular species of hydrogenase in the cells of an obligately chemolithoautotrophic, marine hydrogen-oxidizing bacterium, Hydrogenovibrio marinus.
Hydrogenovibrio marinus was suggested to have only membrane-bound hydrogenase (MBH). The change of cultivation pO2 did not affect the molecular species of hydrogenase expressed. We propose the MBH is grouped in class I [NiFe] MBH according to the subunit composition, size (Mw 38,000 and Mw 74,000 subunits) and N-terminal sequences of the subunits, and arrangement of the structural genes. Ni-requirement for the autotrophic growth on H2 also suggested the MBH is the Ni-containing type. Southern hybridization analysis using a part of the MBH gene showed a possibility of the presence of two highly homologous MBHs which were not separated by SDS-PAGE.


The influence of changing cataract techniques on the ophthalmologist concerned with retinal diseases.
This paper briefly presents a review of some of the retinal problems the ophthalmologist faces because of changing techniques in cataract surgery. It emphasizes the need for controlled clinical trials to properly evaluate the benefits or disadvantages of these changing techniques.


Distribution and chemical coding of phoenixin-immunoreactive nerve structures in the spinal cord of the pig.
Phoenixin (PNX) is a newly described peptide found in both neural and non-neural tissues. Until now, no attempts have been made to investigate the expression of PNX in the nervous system of animals other than laboratory rodents, in which an enzyme immunoassay revealed the highest quantity of the substance in the spinal cord. Since the domestic pig, due to its anatomical and histological resemblance to humans, is often used as an animal model in biomedical investigations, the present study was designed to examine PNX-immunoreactivity in the spinal cords of female pigs (n=5). The spinal cords were dissected and divided into the cervical, thoracic, lumbar, sacral and coccygeal segments, which were sectioned transversally into 10-μm-thick serial sections. The sections from each spinal cord segment were processed for double-labelling immunohistochemistry using antibodies against PNX in a mixture with those against calcitonin gene-related peptide (CGRP), substance P (SP) or choline acetyltransferase (CHAT). The PNX-immunoreactivity had a similar distribution in the grey matter of all the spinal cord sections examined and was mainly observed in varicose nerve fibres (NF) that formed a dense plexus in laminae I and II of the dorsal horn. Nearly all of the PNX-immunoreactive NF stained also for CGRP or SP and, interestingly, many of them were CHAT-positive. The present study has provided for the first time the detailed information on the arrangement and chemical features of nerve structures expressing PNX-immunoreactivity in the spinal cord of a large mammal. The exact function of PNX in the spinal cord is not known yet. However, the distribution pattern and immunohistochemical characteristics of PNX-IR NF clearly suggest that this peptite most likely plays a role in spinal noxious signalling.


Secular trends in weight, weight-for-height and triceps skinfold thickness in primary schoolchildren in England and Scotland from 1972 to 1980.
Measurements of height, weight and triceps skinfold thickness were obtained from English and Scottish schoolchildren from 15 birth cohorts over the period 1972-1980. Positive trends in weight were found for boys and girls in both countries, but in England they were less than expected from the increases in height. The increase in weight-for-height in Scotland was paralleled by a greater increase in triceps skinfold in Scottish than in English children. Scottish children remained shorter, lighter and thinner than English children, but similar in weight-for-height. Separate monitoring of trends in obesity for English and Scottish children should continue.


The Influence of Covid-19 on Perceived Health Effects of Wetland Parks in China.
Wetland parks are designed to support urban ecological protection, flood control and human well-being. Existing research mainly focuses on their influence on ecology and economy. However, their influence on human well-being and health is rarely studied. In China, during the peak of the COVID-19 pandemic (Peak), people were very concerned about health, while at the same time wetland parks which are generally considered beneficial to health were closed. Thus, this study explores the public's perception of the health effects of visiting wetland parks and the impact of the pandemic on the perception. From March 5th to 8th, 2020, before the Peak in China was over, 1,400 respondents participated in a nationwide online survey. It was found that the perceived benefits from visiting wetland parks were higher in terms of mental health than in physical health. Also, the perceived health benefits of wetland parks after the Peak were slightly higher than before the pandemic. The results highlight that wildlife habitat services were considered to be the most important ecosystem services that promote the perceived health benefits. Interestingly, the perceived health benefits of wetland parks by health experts appear to be lower than in other groups, indicating that the health benefits of visiting wetland parks may be overestimated by lay-people or underestimated by health experts. The results provide empirical evidence for managing ecosystem services as delivered by these urban wetlands, in the context of COVID-19 or potential future pandemics, for promoting public health.


The Society of Thoracic Surgeons Adult Cardiac Surgery Database: 2016 Update on Outcomes and Quality.
The Society of Thoracic Surgeons Adult Cardiac Database is one of the longest-standing, largest, and most highly regarded clinical data registries in health care. It serves as the foundation for all quality measurement and improvement activities of The Society of Thoracic Surgeons. This report summarizes current aggregate national outcomes in adult cardiac surgery and reviews database-related activities in the areas of quality measurement and performance improvement.


Treatment of severe Henoch-Schönlein and immunoglobulin A nephritis. A single center experience.
Our aim was to report the effect of two treatment regimens in 43 cases of severe Henoch-Schönlein nephritis (HSN) and immunoglobulin A nephritis (IgAN) (24 HSN, 19 IgAN). Group A, 11 HSN and 7 IgAN, 88% with an International Study of Kidney Disease in Children (ISKDC) biopsy grade > or = III and severe clinical features, were treated with corticosteroids, cyclophosphamide (CYC-P) and angiotensin-converting enzyme inhibitor/angiotensin receptor blocker (ACEi/ARB). Group B, 12 HSN and 13 IgAN, 72% with biopsy findings as above and 52% with severe clinical features, were treated with ACEi/ARB +/- corticosteroids. The outcome classification was: (a) healthy; (b) mild proteinuria, normal glomerular filtration rate (GFR); (c) active renal disease; (d) chronic renal failure. Twenty-six patients had a good outcome (a + b). The 17 children with poor outcome (c + d) had lower GFR at onset and at follow-up, higher albumin excretion at follow-up, and higher percentage of segmental glomerulosclerosis in the renal biopsy, than those with good outcome. Treatment with corticosteroids, CYC-P and ACEi/ARB was effective in increasing GFR, reducing proteinuria and decreasing the disease activity index. The proteinuria had decreased at follow-up in both groups. In group A, GFR increased and histopathological activity index declined after treatment. The outcome did not differ between groups A and B. The effects of treatment did not differ between HSN and IgAN.


The VeA regulatory system and its role in morphological and chemical development in fungi.
In fungi, the velvet gene, or veA, is involved in the regulation of diverse cellular processes, including control of asexual and sexual development as well as secondary metabolism. This global regulator is conserved in numerous fungal species. Interestingly, in Aspergilli, where most of the studies on veA have been carried out, this gene has been described to mediate development in response to light. In recent years the knowledge of this important regulatory system has expanded through the use of Aspergillus nidulans as a model organism, and through the study of veA orthologs across fungal genera. This review includes information on the current understanding of veA function and its mechanism of action. The fact that veA has only been found in fungi, together with advances in the elucidation of the veA mechanism, might be useful in designing future control strategies to decrease the detrimental effects of fungi while enhancing those qualities that are beneficial.


Lymph Node Dissection in Intrahepatic Cholangiocarcinoma: a Critical and Updated Review of the Literature.
Lymphatic spread of intrahepatic cholangiocarcinoma (iCCA) is common and negatively impacts survival. However, the precise role of lymph node dissection (LND) in oncologic outcomes for patients with intrahepatic cholangiocarcinoma remains to be established. Updated evidence on the preoperative diagnosis and prognostic value of lymph node metastasis is reviewed, as well as the potential benefit of LND in patients with iCCA. The ability to accurately determine nodal status for iCCA with current imaging modalities is equivocal. LND has prognostic value for both survival and disease recurrence. However, execution rates of LND are highly varied in the literature, ranging from 26.9 to 100%. At least 6 lymph nodes should be examined from nodal stations of the hepatoduodenal ligament and hepatic artery as well as based on the location of the primary tumor. Neoadjuvant therapies may be beneficial if lymph node metastases at diagnosis are suspected. Surgeons performing a minimally invasive approach should focus on increasing LND rates and harvesting ≥ 6 lymph nodes. Lymph node negativity is required in patients with iCCA being considered for liver transplantation under investigational protocols. Despite an upward trend in the LND rate, the reality is that only 10% of patients with iCCA receive an adequate LND. This review underscores the importance of routinely increasing the rate of adequate LND in these patients in order to achieve accurate staging, appropriately select patients for adjuvant therapy, and improve the prognosis of clinical outcomes. While prospective data is lacking, the therapeutic impact of LND remains unknown.


Transcutaneous carbon dioxide pressure for monitoring patients with severe croup.
In a prospective investigation of 17 children with severe croup, we analyzed the effect of epinephrine inhalations and mild sedation with chloral hydrate on transcutaneous carbon dioxide pressure (tcPCO2), pulse oximetry measurements, and croup scores. There was a highly significant reduction (p less than 0.001) in the tcPCO2 values and croup scores after inhalation of epinephrine. The changes in the tcPCO2 values correlated with the clinical findings. Mild sedation also significantly improved the croup scores but failed to influence the tcPCO2 values. There was not statistically significant difference in pulse oximetry saturation, fraction of administered oxygen, heart rate, or respiratory rate before and after inhalation of epinephrine or chloral hydrate administration. Monitoring tcPCO2 appears to be a reliable and objective tool for managing patients with upper airway obstruction, whereas croup scores may be misleading.


Impact of omeprazole on the plasma clearance of methotrexate.
Omeprazole inhibits the gastric hydrogen pump and is an effective treatment for peptic ulcers. Methotrexate is a chemotherapeutic agent that inhibits dihydrofolate reductase and is eliminated by a hydrogen-ion-dependent mechanism in the kidney. We present evidence that omeprazole inhibits methotrexate clearance and may result in potentially toxic methotrexate levels.


The biosynthesis of 11-ketotestosterone by the testis of the Siamese fighting fish Betta splendens Regan (Anabantoidei, Belontiidae).
Testicular tissues of the Siamese fighting fish were incubated with [14C]pregnenolone for 10, 20, 30, 50, 80, and 120 min, and with [14C]progesterone, [14C]11 beta-hydroxyandrostenedione, [14C]11 beta-hydroxytestosterone, and [14C]androstenetrione for 120 min. 11-Ketotestosterone was the main metabolite in all 120-min incubations. No 11-oxygenated C21 steroids were found as metabolites of either pregnenolone or progesterone. The biosynthesis of 11-ketotestosterone proceeded through both the delta 5- and the delta 4- pathways as judged from the shape of the yield-time curves of the metabolites of pregnenolone. 11-Ketotestosterone formation from 11-oxygenated precursors increased in the order 11 beta-hydroxytestosterone less than 11 beta-hydroxyandrostenedione less than androstenetrione.


Esophageal motor function in Graves' disease.
Esophageal motor function was studied in 10 patients with untreated Graves' disease and 15 healthy volunteers who served as controls. A noncompliant recording system with a triple-lumen assembly was used. Resting upper- and lower-esophageal-sphincter pressures, amplitude, duration, and velocity of peristaltic contractions in the body of the esophagus, and the number of repetitive and simultaneous contractions were measured. There was no significant difference in peristaltic amplitude and duration between the controls and the patients with Graves' disease. However, there was a significant increase in velocity of contractions in Graves' disease as compared with controls. On restudy of 5 patients who became euthyroid after treatment with 131I, all the measured parameters were unchanged except for velocity of the peristaltic contractions. The value for this measurement decreased essentially to the same as that found in the control subjects. It is concluded that: (1) thyrotoxicosis increases the propagation velocity of esophageal contraction, and (2) thyroid hormones may play a physiologic role in the control of esophageal motor function.


Selective sweep of a newly evolved sperm-specific gene in Drosophila.
The pattern of genetic variation across the genome of Drosophila melanogaster is consistent with the occurrence of frequent 'selective sweeps', in which new favourable mutations become incorporated into the species so quickly that linked alleles can 'hitchhike' and also become fixed. Because of the hitchhiking of linked genes, it is generally difficult to identify the target of any putative selective sweep. Here, however, we identify a new gene in D. melanogaster that codes for a sperm-specific axonemal dynein subunit. The gene has a new testes-specific promoter derived from a protein-coding region in a gene encoding the cell-adhesion protein annexin X (AnnX), and it contains a new protein-coding exon derived from an intron in a gene encoding a cytoplasmic dynein intermediate chain (Cdic). The new transcription unit, designated Sdic (for sperm-specific dynein intermediate chain), has been duplicated about tenfold in a tandem array. Consistent with the selective sweep of this gene, the level of genetic polymorphism near Sdic is unusually low. The discovery of this gene supports other results that point to the rapid molecular evolution of male reproductive functions.


Morphological changes due to Lead exposure and the influence of DMSA on the gill tissues of the freshwater fish, Catla catla.
Catla catla fingerlings were reared in freshwater and exposed to 15.5 ppm concentration of lead for 60 days. The morphological changes on the gill of the C. catla fingerlings due to lead intoxication and the effect of DMSA (meso 2,3-dimercaptosuccinic acid) on the affected tissues were observed using Scanning Electron Microscope. It has been found that the lead treated gill tissues showed certain marked changes, such as cell hypertrophy, alteration in the lamellar surfaces, epithelial hyperplasia and the fusion of adjacent lamellae. The antidote DMSA treatment reduces the toxic effects and helps the recovery of gill tissue and its return to the level of the control/normal.


Subperiosteal intraorbital haematoma following minor head trauma. Case report.
Subperiosteal haematoma of the orbit following minor head trauma is extremely rare. A 5-year-old girl is presented with bilateral proptosis, chemosis, extraocular palsy, and progressive visual loss after minor head trauma. She had no evidence of fracture or abnormality of coagulation. The intraorbital haematoma resulted most probably from the oozings of a subgaleal haemorrhage which entered the subperiosteal space and then dissected over the supraorbital ridge into the orbit.


Association of depression symptoms with receipt of healthcare provider advice on physical activity among US adults.
Given the high burden and prevalence of depression, various guidelines underscore the role of healthcare providers in supplying advice on physical activity (PA) as a potential modifying factor influencing the incidence and severity of depressive symptoms in adults. We aimed to investigate the extent to which healthcare providers provide PA advice to adults with depressive symptoms in the US. Data on adults aged 20-64 years (n = 4971) in the National Health and Nutrition Examination Study between 2011 and 2016 were analysed. Depressive symptoms were assessed using the Patient Health Questionnaire and response options were categorised as "none or minimal", "mild", "moderate-severe". Receipt of PA advice from a healthcare provider was self-reported. We restricted our study sample to adults free from chronic diseases. Higher odds of receiving advice to exercise were reported among adults with mild (OR = 1.7, 95% CI: 1.3-2.3) and moderate-severe depressive symptoms (OR = 1.7, 95% CI: 1.0-2.8). Furthermore, exercise advice was more commonly reported among adults who were overweight, obese, Hispanic, Asian, being insured with private insurance, with education higher than high school, and had access to a routine place for health care. Social and culutral aspects of overweight/obesity may prohibit generalizations. Cross sectional design does not allow for causal realtionships. In the US, fewer than one in three adults experiencing symptoms of depression report having received exercise advice from a healthcare provider. Providing such advice may be a sustainable clinical strategy in reducing the incidence and severity of depression symptoms.


Radiation therapy, an important mode of treatment for head and neck chemodectomas.
Between 1970 and 1990, 22 patients with 44 chemodectomas in the head and neck region were seen at the Netherlands Cancer Institute in Amsterdam. All patients were treated with radiation therapy (17 patients with radiation therapy only and 5 in combination with surgery). One patient was treated two times with an interval of 12 years at each side of the neck. Standard dose was 50 Gy in 25 fractions over 5 weeks. A radiation portal arrangement with oblique fields with paired wedges was used most frequently. The follow-up period ranged from 1 year to 20 years. Two recurrences at 2 and 9 years after treatment were observed. The actuarial local control rate was 88% at 10 years follow-up. Comparison of the results of surgery and radiotherapy demonstrates that radiation therapy is an effective treatment modality without mutilation or severe late morbidity for chemodectomas in the head and neck region.


Nucleus accumbens carbachol disrupts olfactory and contextual fear-potentiated startle and attenuates baseline startle reactivity.
Although the nucleus accumbens (NAc) typically is not considered a primary component of the circuitry underlying either the acquisition or retrieval of conditioned fear, evidence suggests that this region may play some role in modulating fear-related behaviors. The goal of the present study was to explore a potential role for NAc cholinergic receptors in the expression of fear-potentiated startle (FPS) and baseline startle reactivity. Intra-NAc infusion of the broad-acting cholinergic receptor agonist, carbachol, suppressed FPS elicited by re-exposure to both a discrete odor previously paired with footshock and the conditioning context. Although carbachol elevated spontaneous motor activity, activity bouts did not account for startle suppression in carbachol-treated Ss. In addition, intra-NAc carbachol suppressed baseline startle over a range of acoustic pulse intensities in the absence of explicit fear conditioning. Collectively, these findings suggest that NAc cholinergic receptors play a role in the modulation of baseline startle reactivity, rather than in the retrieval of learned fear, and that this role is independent of overt motor activity.


Two endogenous protein kinase activities in heterogeneous nuclear ribonucleoprotein particles (hnRNP).
Two protein kinase activities which differ in their catalytic activity towards endogenous and exogenous substrates have been detected and partially purified from heterogeneous nuclear ribonucleoprotein particles (hnRNP).


Regional cerebrovascular reactivity to carbon dioxide during cardiopulmonary bypass in patients with cerebrovascular disease.
In patients with cerebrovascular disease, hypercarbia may cause redistribution of regional cerebral blood flow from marginally perfused to well-perfused regions (intracerebral steal), as evidenced by regional cerebral blood flow studies during carotid endarterectomy. During hypothermic cardiopulmonary bypass, the pH-stat method of acid-base management produces relative hypercarbia. To determine whether pH-stat management produces relative hypercarbia. To determine whether pH-stat management induces intracerebral steals, we investigated nine patients with cerebrovascular disease undergoing coronary artery bypass grafting. During hypothermic cardiopulmonary bypass, arterial carbon dioxide tension was varied in random order between 40 mm Hg and 60 mm Hg (uncorrected for body temperature). Regional cerebral blood flow was measured by clearance of 133 xenon injected into the arterial inflow cannula. Nasopharyngeal temperature (26.8 degrees-28.0 degrees +/- 2.2 degrees-3.0 degrees C), perfusion flow rate (2.14-2.18 +/- 0.70-0.73 L/min/m2), mean arterial pressure (67-68 +/- 6-9 mm Hg), arterial carbon dioxide tension (302-308 +/- 109-113 mm Hg), and hematocrit (23% +/- 4%) were maintained within narrow limits in each patient during arterial carbon dioxide tension manipulation. Global mean cerebral blood flow values were similar to previously reported values in patients free of cerebrovascular disease; patients in this study averaged 15.2 +/- 2.5 ml/100 gm/min at an arterial carbon dioxide tension of 46.1 +/- 8.4 mm Hg and 25.3 +/- 6.1 ml/100 gm/min at an arterial carbon dioxide tension of 71.1 +/- 11.8 mm Hg. Carbon dioxide reactivity, defined as mean global cerebral blood flow (in ml/100 gm/min) divided by arterial carbon dioxide tension (in mm Hg), was similar in the region having the lowest regional cerebral blood flow and in the brain as a whole. No patient developed evidence of an intracerebral steal at the higher arterial carbon dioxide tension. During hypothermic cardiopulmonary bypass, higher levels of arterial carbon dioxide tension, such as those associated with the pH-stat management technique, are apparently not associated with potentially harmful redistribution of cerebral blood flow in patients with cerebrovascular disease.


Absorption and metabolism of the mycotoxins alternariol and alternariol-9-methyl ether in Caco-2 cells in vitro.
Alternariol (AOH) and alternariol-9-methyl ether (AME) are major toxins produced by fungi of the genus Alternaria. In order to simulate their in vivo intestinal absorption and metabolism, AOH and AME have been studied in differentiated Caco-2 cells and in the Caco-2 Millicell® system in vitro. AOH was found to be readily conjugated to two glucuronides and one sulfate, whereas AME gave rise to one major glucuronide and one sulfate. Whereas the glucuronides of AOH and AME were sequestered about equally well into the basolateral and the apical compartment, the sulfates of both toxins were preferentially released to the apical side. Unconjugated AOH but not AME aglycone reached the basolateral chamber. The apparent permeability coefficients (Papp values) were calculated for the aglycones as well as total mycotoxin-associated compounds using an initial apical concentration of 20 µmol/l AOH or AME. Based on these Papp values, AOH must be expected to be extensively and rapidly absorbed from the intestinal lumen in vivo and reach the portal blood both as aglycone and as glucuronide and sulfate. In contrast, intestinal absorption of AME appears to be poor and sluggish, with no AME agylcone and only AME conjugates reaching the portal blood.


Versatile Product Detection via Coupled Assays for Ultrahigh-Throughput Screening of Carbohydrate-Active Enzymes in Microfluidic Droplets.
Enzyme discovery and directed evolution are the two major contemporary approaches for the improvement of industrial processes by biocatalysis in various fields. Customization of catalysts for improvement of single enzyme reactions or de novo reaction development is often complex and tedious. The success of screening campaigns relies on the fraction of sequence space that can be sampled, whether for evolving a particular enzyme or screening metagenomes. Ultrahigh-throughput screening (uHTS) based on in vitro compartmentalization in water-in-oil emulsion of picoliter droplets generated in microfluidic systems allows screening rates >1 kHz (or >107 per day). Screening for carbohydrate-active enzymes (CAZymes) catalyzing biotechnologically valuable reactions in this format presents an additional challenge because the released carbohydrates are difficult to monitor in high throughput. Activated substrates with large optically active hydrophobic leaving groups provide a generic optical readout, but the molecular recognition properties of sugars will be altered by the incorporation of such fluoro- or chromophores and their typically higher reactivity, as leaving groups with lowered pKa values compared to native substrates make the observation of promiscuous reactions more likely. To overcome these issues, we designed microdroplet assays in which optically inactive carbohydrate products are made visible by specific cascades: the primary reaction of an unlabeled substrate leads to an optical signal downstream. Successfully implementing such assays at the picoliter droplet scale allowed us to detect glucose, xylose, glucuronic acid, and arabinose as final products of complex oligosaccharide degradation by glycoside hydrolases by absorbance measurements. Enabling the use of uHTS for screening CAZyme reactions that have been thus far elusive will chart a route toward faster and easier development of specific and efficient biocatalysts for biovalorization, directing enzyme discovery by challenging catalysts for reaction with natural rather than model substrates.


The Impact of Head-Up Tilt Sleeping on Orthostatic Tolerance: A Scoping Review.
To systematically summarize the evidence of head-up tilt sleeping (HUTS) on orthostatic tolerance, we conducted a systematic, predefined search in PubMed, OVID Embase, Cochrane and Web of Science. We included studies assessing the effect of HUTS on orthostatic tolerance and other cardiovascular measures and rated the quality with the American Academy of Neurology risk of bias tool. We included 10 studies (n = 185) in four groups: orthostatic hypotension (OH; 6 studies, n = 103), vasovagal syncope (1 study, n = 12), nocturnal angina pectoris (1 study, n = 10) and healthy subjects (2 studies, n = 58). HUTS duration varied (1 day-4 months) with variable inclinations (5°-15°). In two of six OH studies, HUTS significantly improved standing systolic blood pressure. Orthostatic tolerance was consistently enhanced in OH studies with higher angles (≥12°), in 2 out of 3 with smaller angles (5°) but also in one studying horizontal sleeping. In vasovagal syncope, HUTS significantly augmented resilience to extreme orthostatic stress. One study was rated as a class II risk of bias, one of Class II/III and eight of Class IV. The evidence favouring HUTS to improve orthostatic tolerance is weak due to variable interventions, populations, small samples and a high risk of bias. Despite this, we found some physiological signs suggesting a beneficial effect.


Gene expression profiling of resting and activated vascular smooth muscle cells by serial analysis of gene expression and clustering analysis.
Migration and proliferation of vascular smooth muscle cells (SMCs) are key events in atherosclerosis. However, little is known about alterations in gene expression upon transition of the quiescent, contractile SMC to the proliferative SMC. We performed serial analysis of gene expression (SAGE) of cultured, human SMCs, either grown under resting circumstances or activated with an atherogenic stimulus. Analysis of tags, representing 47,209 and 47,259 mRNAs from a library of resting and activated SMCs, respectively, identified 105 tags induced and 52 tags repressed greater than fivefold. To evaluate the relevance in SMC biology of unmatched, regulated tags, we performed hierarchical clustering analysis, based on their expression profiles in public SAGE databases, and clustered these novel genes in distinct groups. The regulation in SMCs was confirmed by Northern blotting for representative genes of these groups. Plasminogen activator inhibitor-2 has not been associated with atherosclerosis before and was localized to atherosclerotic lesions.


Enzymatic microreactors for the determination of ethanol by an automatic sequential injection analysis system.
A sequential injection analysis system with two enzymatic microreactors for the determination of ethanol has been designed. Alcohol oxidase and horseradish peroxidase were separately immobilized on glass aminopropyl beads, and packed in 0.91-mL volume microreactors, working in line with the sequential injection analysis system. A stop flow of 120 s was selected for a linear ethanol range of 0.005-0.04 g/L +/- 0.6% relative standard deviation with a throughput of seven analyses per hour. The system was applied to measure ethanol concentrations in samples of distilled and nondistilled alcoholic beverages, and of alcoholic fermentation with good performance and no significant difference compared with other analytical procedures (gas chromatography and high-performance liquid chromatography).


C-reactive protein level as a predictor of mortality in liver disease patients with bacteremia.
C-reactive protein (CRP) is synthesized in the liver in response to inflammation, and CRP is a widely used marker of sepsis. In bacteremia the initial CRP level is an independent predictor of mortality. Since the CRP response in patients with chronic liver disease is lower than in patients without liver disease the objective was to assess whether CRP levels in chronic liver disease and bacteremia was associated with case fatality. The study enrolled 105 patients with chronic liver disease and bacteremia as well as 202 patients with bacteremia and no recorded liver disease from the same region and time period. Retrospective review of medical records with registration of demography, co-morbidity, bacteriological, biochemical and clinical findings, and Child-Turcotte-Pugh scores. The primary outcome was 30-day mortality. Mortality was significantly higher in patients with chronic liver disease (mortality rate ratio 2.2; 95% confidence interval 1.2-3.9) and it was correlated to Child-Turcotte-Pugh scores. CRP levels were not different between the three Child-Turcotte-Pugh classes (p = 0.33), and no linear correlation with 30-day mortality was observed. Mortality associated with bacteremia is increased in patients with chronic liver disease and it is correlated with Child-Turcotte-Pugh score. The prognostic information of initial CRP levels in patients with chronic liver disease is weak. The clinical management of patients with chronic liver disease and suspected infection should initiate antimicrobial therapy based on clinical, radiological and microbiological findings, whereas the measurement of CRP in bacteremia is less helpful as compared with patients without liver disease.


Setup uncertainties for inter-fractional head and neck cancer in radiotherapy.
The aim of this study is to determine the inter-fractional motion of cervical spine in radiotherapy (RT). Eleven localized head and neck cancer patients who were treated from April 2014 to September 2015 were evaluated. Every patient underwent 3 times of computed tomography (CT) simulation with equivalent setting. Left-right (LR, x) and antero-posterior (AP, z) directional shift of cervical spine were evaluated using 33 number of CT image. In regard to random error, geometric changes were evaluated by 22 data set (compared the first obtained CT to second or third CT) by one-sample T test. Systemic error was evaluated by each patients' data set (11 pairs) by paired T test. The mean random error of LR and AP translational shift of cervical spine were -0.39 ± 3.24 mm and -0.57 ± 0.99 mm respectively. The mean random error of translational change of AP direction showed statistical significance (p = 0.014). The mean random error of x and z rotational shift were -0.07 ± 0.29° and -0.05 ± 0.35°, respectively. The mean systemic error of translational shift of LR and AP direction were -0.64 ± 2.57 mm and -0.33 ± 1.22 mm, respectively. The mean systemic error of rotational shift of x and z were 0.01 ± 0.18° and -0.27 ± 0.33°, respectively. The mean systemic error of rotational changes of z direction showed statistical difference (p = 0.022). We have to be aware of the inter-fractional motion of cervical spine in head and neck RT and give enough margins in RT planning.


Genetic epidemiology of an institutionalized cohort of mental retardates.
By criterion scaling and principal component analysis of performance, social class, symptoms, institutionalization, and medical history, a cohort of mental defectives has been divided into medical, biological, and sociofamilial categories. This division, corresponding to differences in etiology and severity, reveals changing patterns of admission and provides evidence that male excess is not primarily due to sex linkage. The incidence of mental retardation increases about 5% with first-cousin marriage, in agreement with other studies. The decline of IQ with inbreeding appears to be due entirely to rare recessive genes, not to dominance deviations of polygenes.


Preliminary studies on the composition and rheological properties of the extracellular polysaccharide synthesized by Pseudomonas PBI (NCIB 11264).
A microorganism isolated from a carbohydrate-rich industrial effluent synthesized an exocellular slime polysaccharide composed of glucose and galactose in a molar ratio of 7.45 +/- 0.68 : 1, and two non-carbohydrate substituents acetate (3--4%) and pyruvate (5--9%). Contamination by rhamnose and mannose was detectable in crude polysaccharide samples. Solutions of the polysaccharide were pseudoplastic, but not thixotropic, and formed gels in the presence of certain trivalent cations.


Enhanced degradation of few-layer black phosphorus by fulvic acid: Processes and mechanisms.
The oxidation of the emerging nanomaterial black phosphorus (BP) affected by pH and oxygen has been carefully documented. However, in natural waters, there is a large amount of chemically reactive organic matters like fulvic acid (FA), whose impacts on degradation and stability of few-layer BP or BP nanosheets (BPNS) are scarcely disclosed. Hence, we investigated the kinetics of BPNS degradation products (H2PO2-, HPO32-, and PO43-) in the presence of FA. The results showed that the apparent reaction rate constants of BPNS were 0.026, 0.050, and 0.060 d-1 under oxygen-and-light condition and 0.005, 0.016, and 0.023 d-1 under hypoxia-and-darkness condition at FA gradients of 0, 2.5, and 5 mgC/L, respectively. Microscopic observations, simple molecular simulation experiment, and density functional theory computation explained that FA significantly enhanced the degradation of P atoms on the BPNS surface through the indirect pathway of reducing the energy barrier of O2 dissociative adsorption and the direct pathway of chemical adsorption, which caused the P-P bond on the BPNS surface to break down and formed P-O bonds or C-P bonds. This study revealed for the first time the degradation mechanism of BPNS in the presence of FA, which is a chemical mechanism of the BPNS transformation behavior. It helps to make a more scientific risk assessment of BP in natural waters.


Studies on the blood of a Co (a-b-) proposita and her family.
The serum of a pregnant woman whose red cells typed as Co(a-b-) contained an alloantibody that, at the time of the infants delivery, was hemolytic in vitro and had a titer of 32,000 by the antiglobulin test. This antibody, which reacted with cells of all Colton-phenotypes except the proposita's own cells, showed very weak reactions with other Co(a-b-) cells and therefore cannot be called anti-Co3. The red cells of the proposita may carry a very weak Co3 antigen. No other persons of the Co(a-b-) phenotype were found in her members, among three of the four generations tested, suggested that an inhibitor gene may be responsible for the unusual Colton phenotypes in this family. The proposita's infant required one exchange transfusion with red cells obtained from the proposita. Red cells from 40,000 donors reacted with the serum of the proposita.


The effects of nerve transection on the endoneurial collagen fibril sheaths.
The connective tissue changes during Wallerian degeneration and subsequent regeneration were studied in the distal stump of transected sciatic nerves of Wistar rats. In half of the animals regeneration was prevented by suturing the distal stump to muscle and in the rest spontaneous regeneration was allowed. Intact contralateral nerves served as controls. By 4 weeks after transection the Schwann cell columns became surrounded by a layer of thin collagen fibrils that were, on average, 25-30 nm in diameter. This was only half of the fibril diameter observed elsewhere in the endoneurium or in control nerves. The layer of thin fibrils diminished in thickness when axonal regeneration reached the distal stump, especially as the axons became myelinated. At all stages of the experiment the fibril diameter distribution in the surrounding normal endoneurial stroma was comparable with that observed in control nerves. Segments of Schwann cell basement membrane were observed to be closely associated with collagen fibrils both in freely regenerating, as well as in non-regenerating, nerves. The diameter of these fibrils corresponded to that observed in the zone of thin fibrils surrounding the Schwann cell columns. Such areas were not found in control nerves. The data obtained show that deposition of thin collagen fibrils occurs around the Schwann cell columns as a reaction to transection. Our observations on the regenerating nerves indicate that this connective tissue reaction does not prevent regeneration in the early phases following injury and that its progression is limited concomitantly with axonal regeneration.


Growth and antigenic properties of a spontaneously regressing subline of leukemia L1210.
A subline (1210/MR) of the lethal parental L1210 leukemia cell line has been demonstrated to undergo dose-dependent spontaneous regression in both syngeneic and hybrid mice, although the regressing and non-regressing cells differed only slightly in growth rate. The regressed mice, like those actively immunized, were specifically protected from the subsequent challenge with L1210 cells. In addition, L1210/MR cells when coinjected could protect CD2F1 mice from low but lethal doses of L1210 cells. These two cell lines shared at least one cross-reacting cell surface antigen as determined by an isologous anti-L1210/MR immune serum. Such immune serum after being absorbed by L1210 cells still retained its in vitro complement-dependent cytotoxicity against L1210/MR cells. We conclude that the cell lines are antigenically related but L1210/MR cells may possess additional novel antigens, and that spontaneous regression of L1210 may be an L1210-specific immunological phenomenon.


Motor demands influence conflict processing in a mouse-tracking Simon task.
Previous studies have shown incorrect motor activation when making perceptual decisions under conflict, but the potential involvement of motor processes in conflict resolution is still unclear. The present study tested whether the effects of distracting information may be reduced when anticipated motor processing demands increase. Specifically, across two mouse-tracking Simon experiments, we manipulated blockwise motor demands (high vs. low) by requiring participants to move a mouse cursor to either large versus small (Experiment 1) or near versus far (Experiment 2) response boxes presented on the screen. We reasoned that participants would increase action control in blocks with high versus low motor demands and that this would reduce the distracting effect of location-based activation. The results support this hypothesis: Simon effects were reduced under high versus low motor demands and this modulation held even when controlling for time-varying fluctuations in distractor-based activation via distributional analyses (i.e., delta plots). Thus, the present findings indicate that anticipation of different motor costs can influence conflict processing. We propose that the competition between distractor-based and target-based activation is biased at premotor and/or motor stages in anticipation of motor demands, but also discuss alternative implementations of action control.


[Pelvic exenteration as a means of improving the resectability of advanced cancer of the rectum].
The paper discusses the results of treatment of 10 male patients who underwent pelvic evisceration for advanced rectal cancer. Surgical technique is described. In 9 cases, an isolated ileocecal "urinary bladder" was formed; in 1 patient urine was passed into a small intestine segment. The involvement of the urinary bladder and/or the prostate were observed in 6 patients. One patient died shortly after surgery, three--within the first 12 months and another 2--during the second year. Four cases have survived over 5 years, 3 of them surviving over 10 years.


Genomic structure of the gene for the SH2 and pleckstrin homology domain-containing protein GRB10 and evaluation of its role in Hirschsprung disease.
Hirschsprung disease (HSCR), or congenital aganglionic megacolon, is the most frequent cause of congenital bowel obstruction. Germline mutations in the RET receptor tyrosine kinase have been shown to cause HSCR. Mice that carry null alleles for RET or for its ligand, glial cell line-derived neurotrophic factor (GDNF), both exhibit complete intestinal aganglionosis and renal defects. Recently, the Src homology 2 (SH2) domain-containing protein Grb10 has been shown to interact with RET in vitro and in vivo, early in development. We have confirmed the map location of GRB10 on human chromosome 7, isolated human BACs containing the gene, elucidated its genomic structure, isolated a highly polymorphic microsatellite marker adjacent to exon 14 and scanned the gene for mutations in a large panel of HSCR patients. No evidence of linkage was detected in HSCR kindreds and no mutations were found in patients. These data suggest that while GRB10 may be important for signal transduction in developing embryos, it does not play an obvious role in HSCR.


[Multidisciplinary management in the first few hours after treatment of a stroke].
The first 24 hours of post-treatment management of stroke in a neurovascular unit are critical. Combined assistance from nurses, nursing assistants and physiotherapists ensures patient safety, preventing complications and recurrence and allowing prompt initiation of rehabilitation therapy. Multidisciplinary management focuses on patients and their specific needs based on values shared by all members of the team.


Symptomatic Hallucal Interphalangeal Sesamoid Bones Successfully Treated with Ultrasound-guided Injection - A Case Report -.
The hallucal interphalangeal sesamoid bone is usually asymptomatic, but it is not uncommon for it to be symptomatic in cases of undue pressure, overuse, or trauma. Even in symptomatic cases, however, patients often suffer for extended periods due to misdiagnosis, resulting in depression and anxiety that can steadily worsen to the extent that symptoms are sometimes mistaken for a somatoform disorder. Dynamic ultrasound-guided evaluations can be an effective means of detecting symptomatic sesamoid bones, and a simple injection of a small dose of local anesthetics mixed with steroids is an easily performed and effective treatment option in cases, for example, of tenosynovitis.


Competitive reversals and environment-dependent resource partitioning in Erodium.
Effects of drought and varying plant density on the competitive coexistence of two winter annual Erodium species were studied using multiple regression analysis. Significant indications of resource partitioning were detected for interspecific mixtures under spring drought. Competitive superiority also was environment-dependent with E. botrys dominating with drought in autumn, while E. brachycarpum dominated with drought in spring. The results suggest that competitive coexistence in Erodium is promoted by processes both equilibrial (e.g., resource partitioning) and nonequilibrial (e.g. competitive reversals).


Deterministic early endosomal maturations emerge from a stochastic trigger-and-convert mechanism.
Endosomal maturation is critical for robust and timely cargo transport to specific cellular compartments. The most prominent model of early endosomal maturation involves a phosphoinositide-driven gain or loss of specific proteins on individual endosomes, emphasising an autonomous and stochastic description. However, limitations in fast, volumetric imaging long hindered direct whole cell-level measurements of absolute numbers of maturation events. Here, we use lattice light-sheet imaging and bespoke automated analysis to track individual very early (APPL1-positive) and early (EEA1-positive) endosomes over the entire population, demonstrating that direct inter-endosomal contact drives maturation between these populations. Using fluorescence lifetime, we show that this endosomal interaction is underpinned by asymmetric binding of EEA1 to very early and early endosomes through its N- and C-termini, respectively. In combination with agent-based simulation which supports a 'trigger-and-convert' model, our findings indicate that APPL1- to EEA1-positive maturation is driven not by autonomous events but by heterotypic EEA1-mediated interactions, providing a mechanism for temporal and population-level control of maturation.


Correlation of SV2C rs1423099 single nucleotide polymorphism with sporadic Parkinson's disease in Han population in Southern China.
The synaptic vesicle glycoprotein 2 (SV2) has been implicated in synaptic function throughout the brain. Accumulating evidence investigated that SV2C contributed to dopamine release and the disrupted expression of SV2C was considered to be a unique feature of PD that may facilitate dopaminergic neuron dysfunction. This study aimed to examine the relationship between the SV2C rs1423099 single nucleotide polymorphism and sporadic Parkinson's disease (PD) in the Chinese Han population. This study enrolled 351 patients with sporadic PD and 240 normal controls in Chinese Han population. Peripheral blood DNA was extracted by DNA extraction kits and the rs1423099 genotype was analyzed by Agena MassARRAY DNA mass spectrometry. The differences in genotype and allele distribution frequencies between PD patients and control groups were compared using chi-squared tests or Fisher's exact tests. No statistical difference was revealed in age and sex distribution between the cases and control groups, and the distribution of genotype and allele frequencies was consistent with the Hardy-Weinberg equilibrium test. In SV2C rs1423099 dominant model, the frequency of the CC/CT genotype was significantly higher in the PD group compared to the control group (OR = 4.065,95% CI: 2.801-10.870, p = 0.002). Nevertheless, in the recessive model, CC or CT/TT genotypes have no statistical difference in the two groups (p = 0.09). Additionally, in allelic analysis, the C allele was investigated to increase the risk of PD (OR = 1.346, 95% CI: 1.036-1.745, p = 0.026); Furthermore, subgroup analysis suggested that those carrying the C allele in the male subgroup were at a higher risk to afflicted with PD (OR = 1.637, 95% CI: 1.147-2.336, p = 0.006). SV2C rs1423099 single nucleotide polymorphism was associated with sporadic Parkinson's disease in the Chinese Han population, particularly in males.


A D-pathway mutation decouples the Paracoccus denitrificans cytochrome c oxidase by altering the side-chain orientation of a distant conserved glutamate.
Asparagine 131, located near the cytoplasmic entrance of the D-pathway in subunit I of the Paracoccus denitrificans aa(3) cytochrome c oxidase, is a residue crucial for proton pumping. When replaced by an aspartate, the mutant enzyme is completely decoupled: while retaining full cytochrome c oxidation activity, it does not pump protons. The same phenotype is observed for two other substitutions at this position (N131E and N131C), whereas a conservative replacement by glutamine affects both activities of the enzyme. The N131D variant oxidase was crystallized and its structure was solved to 2.32-A resolution, revealing no significant overall change in the protein structure when compared with the wild type (WT), except for an alternative orientation of the E278 side chain in addition to its WT conformation. Moreover, remarkable differences in the crystallographically resolved chain of water molecules in the D-pathway are found for the variant: four water molecules that are observed in the water chain between N131 and E278 in the WT structure are not visible in the variant, indicating a higher mobility of these water molecules. Electrochemically induced Fourier transform infrared difference spectra of decoupled mutants confirm that the protonation state of E278 is unaltered by these mutations but indicate a distinct perturbation in the hydrogen-bonding environment of this residue. Furthermore, they suggest that the carboxylate side chain of the N131D mutant is deprotonated. These findings are discussed in terms of their mechanistic implications for proton routing through the D-pathway of cytochrome c oxidase.


The potential for QT prolongation by antiepileptic drugs in children.
Cardiac arrhythmia may be one of the major causes of sudden unexpected death in children with epilepsy. We assessed drug-induced QT prolongation to establish whether the use of antiepileptic drugs contributes to sudden unexpected death. A total of 178 children with epilepsy (93 males and 85 females, with ages ranging from 1 month to 18.9 years; mean age 7.0 +/- 4.1 years) were involved in the study. The QT intervals were manually measured and corrected using Fridericia's formula (QTFc = QT/RR(1/3)). The mean corrected QT interval (QTc) of 152 children on antiepileptic drugs during the study period was 0.40 +/- 0.03 s, and for 26 age-matched, antiepileptic drug-free control patients it was 0.40 +/- 0.03 s. The mean QTc of the children with monotherapy was 0.40 +/- 0.03 s for the valproate group (n = 42), 0.39 +/- 0.02 s for the carbamazepine/oxcarbazepine group (n = 34), and 0.40 +/- 0.02 s for the topiramate group (n = 26), respectively. There was no statistically significant difference among the groups as assessed by analysis of variance. In addition, there was no significant difference between the monotherapy group (n = 109; 0.40 +/- 0.02 s) and the polytherapy group (n = 43; 0.39 +/- 0.03 s). Major antiepileptic drugs may not precipitate prolongation of the QT interval into sudden unexpected death in children with epilepsy, however further studies are required.


Agricultural residues for cellulolytic enzyme production by Aspergillus niger: effects of pretreatment.
Different agricultural residues were considered in this study for their ability to support cellulolytic enzyme production by Aspergillus niger. A total of eleven agricultural residues including finger millet hulls, sorghum hulls, soybean hulls, groundnut husk, banana peels, corn stalk, cassava peels, sugarcane bagasse, saw dust, rice straw and sheanut cake were subjected to three pretreatment (acid, alkali and oxidative) methods. All the residues supported the growth and production of cellulases by A. niger after 96 h of incubation. Maximum cellulase production was found in alkali-treated soybean hulls with CMCase, FPase and β-glucosidase yields of 9.91 ± 0.04, 6.20 ± 0.13 and 5.69 ± 0.29 U/g, respectively. Further studies in assessing the potential of soybean hulls are being considered to optimize the medium composition and process parameters for enhanced cellulase production.


Pharmacokinetics and bioequivalence evaluation of acamprosate calcium tablets in healthy Chinese volunteers.
Few pharmacokinetic data of acamprosate were available in Chinese population and no medication is approved for alcohol dependence in China. 1. Investigate the pharmacokinetic properties of acamprosate calcium in healthy Chinese male volunteers on single- and multiple-dose administration. 2. Compare the bioequivalence of two formulations of acamprosate calcium tablets both under fasting and fed conditions. This open-label, randomized study included 3 stages. In each stage, a 2-way crossover bioequivalence study was conducted to study the pharmacokinetic properties and bioequivalence of acamprosate calcium tablets on multiple dosing after standardized meals, single dosing under fasting conditions and fed conditions, respectively. The washout period between each treatment in a stage and between each stage was 1week. Plasma acamprosate calcium was quantified by a validated liquid chromatography-tandem mass spectrometry (LC-MS/MS) method. Tolerability was evaluated by monitoring adverse events, physical examinations, 12-lead ECG, and laboratory tests. Totally, 36 male subjects were enrolled in the study and all of them completed the whole 3 study stages. Main pharmacokinetic parameters of test and reference formulations were as follows: multiple dosing, Tmax 9.94±6.59 and 9.47±5.47h, Cmax 435.74±348.10 and 346.54±155.66ng·mL(-1), AUC0-t 8600.52±5264.77 and 9315.10±6820.03ng·mL(-1)·h, AUC0-∞ 8845.38±5838.18 and 9669.24±7326.53ng·mL(-1)·h, t1/2 10.06±8.83 and 9.87±10.35h; single dosing under fasting conditions, Tmax 7.29±4.87 and 6.57±1.85h, Cmax 247.85±110.05 and 244.64±132.43ng·mL(-1), AUC0-t 3385.41±1418.92 and 3496.24±1767.29ng·mL(-1)·h, AUC0-∞ 3781.53±1556.96 and 3829.56±1981.25ng·mL(-1)·h, t1/2 13.07±17.24 and 10.26±7.78h; single dosing under fed conditions, Tmax 17.72±9.42 and 19.50±9.84h, Cmax 183.90±74.52 and 168.14±60.67ng·mL(-1), AUC0-t 3181.71±1368.24 and 3575.11±1416.39ng·mL(-1)·h, AUC0-∞3442.39±2002.53 and 3624.44±1418.12ng·mL(-1)·h, t1/2 8.76±12.28 and 6.67±4.84h, respectively. In all three stages, 90% CIs for the test/reference ratio of AUC0-t and AUC0-∞ were located within 80%-125%, 90% CI for Cmax was within 70%-143%. Similar pharmacokinetic results of acamprosate calcium tablets in healthy Chinese volunteers were found as those in Caucasic population. In all three stages, the two formulations met the regulatory criteria for bioequivalence. Chictr.org identifier: ChiCTR-TTRCC-14004853.


Small changes approach promotes initial and continued weight loss with a phone-based follow-up: nine-month outcomes from ASPIRES II.
To examine the impact of a small-changes weight loss program across a 3-month intervention followed by a 6-month follow-up program. A one-group pre-post intervention study. Medium-sized Southwestern university. Twenty-five obese adult women (mean body mass index [BMI] = 31.8 kg/m(2), standard deviation [SD] = 4.9). Participants were asked to choose and adopt small changes in their diet and physical activity relative to baseline during weekly group-based meetings over 3 months. Participants then received bi-weekly phone calls across a 6-month follow-up period. Weight change was the primary outcome. Secondary outcomes included waist circumference, daily step count, and caloric intake. Intention-to-treat analysis of change from baseline and completers-only analysis (n = 22) for secondary outcomes. Participants achieved clinically significant weight loss (mean [M] = -3.2 kg, standard error [SE] = .47 kg, p < .001) across the initial small changes treatment program. Moreover, participants continued to lose weight across the 6-month phone-based follow-up program (M = -2.1 kg, SE = .83 kg, p < .017), totaling >5% weight loss across the 9-month program (M = 5.3 kg, SE = 1.1 kg, p < .001). Using a small changes approach, participants achieved weight loss in an initial group-based program, which continued with minimal phone-based follow-up. Larger randomized studies comparing a small changes approach to traditional obesity treatment are warranted.


Authors' response to the commentary by Bonaventura and Montecucco on: 'Characterization of circulating leukocytes and correlation of leukocyte subsets with metabolic parameters 1 and 5 years after diabetes diagnosis'.


[Purification and characteristics of methanol dehydrogenase of Methylobacterium nodulans rhizosphere phytosymbionts].
Methanol dehydrogenase (MDG) of the facultative methylotrophic phytosymbiont Methylobacterium nodulans has been purified for the first time to an electrophoretically homogeneous state and characterized. The native protein with a molecular mass of 70 kDa consists of large (60 kDa) and small (6 kDa) subunits. The purified protein displayed a specter identical to that of pyrochinolinchinon (PCC)-containing MDGs (pI 8.7, pH optimum in the range 9-10). The enzyme was inactive in the absence of ammonium or methylamine and exhibited a wide substrate specificity with regard to C1-C2 alcohols with the highest affinity to methanol (K(M) = 70 mM), but it did not oxidize benzyl and secondary alcohols. The apparent values of K(M) to primary alcohols increased with the length of the carbonic chain. The enzyme was characterized by a high stability level even in the absence of a substrate. An immobilized enzyme was used for amperometric methanol detection.


Construction of a native promoter-containing transposon vector for the stable monitoring of the denitrifying bacterium Pseudomonas stutzeri LYS-86 by chromosomal-integrated gfp.
Green fluorescent protein (GFP) is the most potential useful marker for the in situ monitoring of biofilm microbes. The objective of this study was to construct and compare the efficacy of transposon vectors containing native and foreign promoters in monitoring the denitrifying bacterium Pseudomonas stutzeri LYS-86 by chromosomal-integrated gfp. The promoter of nitrite reductase (Pnir) was cloned from LYS-86 and utilized to construct the transposon vector pUT/mini-Tn5-km2-Pnir-gfp. Another transposon vector, pUT/mini-Tn5-km2-Plac-gfp, containing the lactose promoter Plac was also constructed. These two transposon vectors and pUT-luxAB-gfp containing the promoter PpsbA were individually inserted into the chromosome of P. stutzeri LYS-86 by conjugation. Three GFP-tagged recombinant strains, LYS-Plac-gfp, LYS-Pnir-gfp, and LYS-PpsbA-gfp, were selected from the conjugants. Green fluorescence was observed only in LYS-Pnir-gfp, suggesting that the native promoter Pnir may be more suitable for GFP expression in P. stutzeri than the foreign promoters Plac and PpsbA. Indeed, LYS-Pnir-gfp maintained stable GFP fluorescence over 16 subcultures without significant changes in the denitrifying capacity.


Total Tumor Load to assist in the decision for additional axillary surgery in the positive sentinel node breast cancer patients.
The Total Tumor Load (TTL) concept has been demonstrated to accurately predict the status of the non-sentinel lymph nodes (NSLN) in breast cancer patients. In 2019, our center implemented the TTL cut-off of 30,000 CK19 mRNA copies/μL as sole criterion for deciding on performing ALND. This retrospective, unicentric, study analyzed 87 cT1-3N0 breast cancer patients treated consecutively in a period of two years and aimed to evaluate the performance of this criterion. Secondary objectives included the comparison of the criterion versus our previous Clinical Decision Rule (CDR) versus ACOSOG Z0011 criteria for avoiding an ALND in proportion of patients spared an ALND and in proportion of patients left with a surgically untreated metastasized axilla. An interim analysis revealed new TTL cut-offs for deciding on performing an ALND. The 30,000 CK19 mRNA copies/μL criterion yielded an area under the ROC Curve (AUC) of 0.849, a false positive (FP) rate of 30.1% and a positive predictive value (PPV) of 38.9%. The 30,000 CK19 mRNA copies/μL criterion spared 58.6% of the patients an ALND versus 41.4% with CDR versus 73.6% with Z0011 and left 0.0% patients with a surgically untreated metastasized axilla versus 21.4% with CDR versus 42.9% with Z0011. The new TTL cut-off of 260,000 CK19 mRNA copies/μL for deciding on an ALND yielded an AUC of 0.753, a FP rate of 13.7% and a PPV of 47.4%. This new criterion spared 78.2% of the study sample an ALND and left 35.7% of metastasized axillae surgically untreated. This study emphasizes the need to find a new balance between locoregional control and the morbidity associated with Berg levels I + II axillary lymph node dissection.


Patchiness in patterns of growth and survival of two grasses.
Individuals of two perennial bunch grasses, Bouteloua rigidiseta and Aristida longiseta, were transplanted into otherwise undisturbed natural short-grass vegetation in which these species are dominants. Significant differences in survivorship and growth rate were found among quadrats for both species. The relative favorability of quadrats for Bouteloua, but not for Aristida, changed during the course of the two year study. For neither species was a consistent relationship between pre-existing density and transplant performance found. However, one patch type was identified in which Bouteloua transplants grow well, although this species is not naturally present there, apparently because it cannot establish. Microsite differences, created by different soil in transplanted peat pots, were also found to be important. Bouteloua had a higher survivorship the first year, Aristida the second, which suggests that Aristida is the more tolerant of drier years.


Macrophage-targeting gene silencing orchestrates myocardial microenvironment remodeling toward the anti-inflammatory treatment of ischemia-reperfusion (IR) injury.
Ischemia-reperfusion (IR) injury represents a major cause of myocardial dysfunction after infarction and thrombolytic therapy, and it is closely related to the free radical explosion and overwhelming inflammatory responses. Herein, macrophage-targeting nanocomplexes (NCs) are developed to mediate efficient co-delivery of siRNA against MOF (siMOF) and microRNA-21 (miR21) into myocardial macrophages, cooperatively orchestrating the myocardial microenvironment against IR injury. Bioreducible, branched poly(β-amino ester) (BPAE-SS) is designed to co-condense siMOF and miR21 into NCs in a multivalency-reinforced approach, and they are surface-decorated with carboxylated mannan (Man-COOH) to shield the positive surface charges and enhance the serum stability. The final MBSsm NCs are efficiently internalized by myocardial macrophages after systemic administration, wherein BPAE-SS is degraded into small segments by intracellular glutathione to promote the siMOF/miR21 release, finally provoking efficient gene silencing. Thus, cardiomyocyte protection and macrophage modulation are realized via the combined effects of ROS scavenging, inflammation inhibition, and autophagy attenuation, which ameliorates the myocardial microenvironment and restores the cardiac function via positive cellular crosstalk. This study renders promising solutions to address the multiple systemic barriers against in vivo nucleic acid delivery, and it also offers new options for IR injury by manipulating multiple reciprocal bio-reactions.


Laparoscopic vs open living donor nephrectomy: a contemporary series from one centre.
To report our experience of laparoscopic living donor nephrectomy (LDN) vs open donor nephrectomy (ODN), as LDN offers potential advantages to the donor and has become a routine procedure for live kidney procurement worldwide. Between February 2000 and August 2005 we performed 183 donor-recipient operations at our institution (ODN, 83; LDN, 100). We prospectively collected information on all donors and recipients for the same period to audit our experience with the first 100 LDNs. Patients made their operative choice after discussions that included unit experience and published information. We present our findings with the emphasis on donor operative details and early recipient graft outcome. Donor and recipient age, gender, body mass index, human leukocyte antigen mismatches, and vascular anastomotic times did not differ significantly between the groups. There were two conversions to an open operation in the LND group; neither affected recipient-graft outcome. The mean (sd) operative duration was 178 (38) min for the LDN and 159 (34) min for the ODN (P < 0.05). The mean (sd) hospital stay was 4.7 (1.2) days in the LND group and 6.8 (1.5) days in the ODN group (P < 0.05). There was one case of delayed graft function in both groups. Serum creatinine levels at 1, 6 and 12 months after transplantation did not differ significantly between the groups. Our contemporaneous series shows the safe introduction of a laparoscopic living-donor programme without compromising donor patient safety or allograft outcome.


[A micro-computer program for statistical analysis of data from twin studies].
A program of statistical processing of data obtained while examining twins is described. Transformed algorithms, directions for programme use and a control model are presented.


The leucine 10 residue in the pleckstrin homology domain of ceramide kinase is crucial for its catalytic activity.
Ceramide kinase (CERK) converts ceramide (Cer) to ceramide-1-phosphate (C1P), a newly recognized bioactive molecule capable of regulating diverse cellular functions. The N-terminus of the CERK protein encompasses a sequence motif known as a pleckstrin homology (PH) domain. However, little is known regarding the functional roles of this domain in CERK. In this study, we have demonstrated that the PH domain of CERK is essential for its enzyme activity. Using site-directed mutagenesis, we have further determined that Leu10 in the PH domain has an important role in CERK activity. Replacing this residue with a neutral alanine or isoleucine, caused a dramatic decrease in CERK activity to 1% and 29%, respectively, compared to CERK, but had no effect on substrate affinity. The study presented here suggests that the PH domain of CERK is not only indispensable for its activity but also act as a regulator of CERK activity.


Crosstalk between Noncoding RNAs and the Epigenetics Machinery in Pediatric Tumors and Their Microenvironment.
According to the World Health Organization, every year, an estimated 400,000+ new cancer cases affect children under the age of 20 worldwide. Unlike adult cancers, pediatric cancers develop very early in life due to alterations in signaling pathways that regulate embryonic development, and environmental factors do not contribute much to cancer development. The highly organized complex microenvironment controlled by synchronized gene expression patterns plays an essential role in the embryonic stages of development. Dysregulated development can lead to tumor initiation and growth. The low mutational burden in pediatric tumors suggests the predominant role of epigenetic changes in driving the cancer phenotype. However, one more upstream layer of regulation driven by ncRNAs regulates gene expression and signaling pathways involved in the development. Deregulation of ncRNAs can alter the epigenetic machinery of a cell, affecting the transcription and translation profiles of gene regulatory networks required for cellular proliferation and differentiation during embryonic development. Therefore, it is essential to understand the role of ncRNAs in pediatric tumor development to accelerate translational research to discover new treatments for childhood cancers. This review focuses on the role of ncRNA in regulating the epigenetics of pediatric tumors and their tumor microenvironment, the impact of their deregulation on driving pediatric tumor progress, and their potential as effective therapeutic targets.