Developmental nicotine exposure alters glycinergic neurotransmission to hypoglossal motoneurons in neonatal rats.
We tested the hypothesis that nicotine exposure in utero and after birth [developmental nicotine exposure (DNE)] disrupts development of glycinergic synaptic transmission to hypoglossal motoneurons (XIIMNs). Glycinergic spontaneous and miniature inhibitory postsynaptic currents (sIPSC/mIPSC) were recorded from XIIMNs in brain stem slices from 1- to 5-day-old rat pups of either sex, under baseline conditions and following stimulation of nicotinic acetylcholine (ACh) receptors with nicotine (i.e., an acute nicotine challenge). Under baseline conditions, there were no significant effects of DNE on the amplitude or frequency of either sIPSCs or mIPSCs. In addition, DNE did not alter the magnitude of the whole cell current evoked by bath application of glycine, consistent with an absence of change in postsynaptic glycine-mediated conductance. An acute nicotine challenge (bath application of 0.5 μM nicotine) increased sIPSC frequency in the DNE cells, but not control cells. In contrast, nicotine challenge did not change mIPSC frequency in either control or DNE cells. In addition, there were no significant changes in the amplitude of either sIPSCs or mIPSCs in response to nicotine challenge. The increased frequency of sIPSCs in response to an acute nicotine challenge in DNE cells reflects an enhancement of action potential-mediated input from glycinergic interneurons to hypoglossal motoneurons. This could lead to more intense inhibition of hypoglossal motoneurons in response to exogenous nicotine or endogenous ACh. The former would occur with smoking or e-cigarette use while the latter occurs with changes in sleep state and with hypercapnia. NEW & NOTEWORTHY Here we show that perinatal nicotine exposure does not impact baseline glycinergic neurotransmission to hypoglossal motoneurons but enhances glycinergic inputs to hypoglossal motoneurons in response to activation of nicotinic acetylcholine (ACh) receptors with acute nicotine. Given that ACh is the endogenous ligand for nicotinic ACh receptors, the latter reveals a potential mechanism whereby perinatal nicotine exposure alters motor function under conditions where ACh release increases, such as the transition from non-rapid-eye movement to rapid-eye movement sleep, and during hypercapnia.


Doppler Ultrasound Monitoring of Echogenicity in Asymptomatic Subcritical Carotid Stenosis and Assessment of Response to Oral Supplementation of Vitamin K2 (PLAK2 Randomized Controlled Trial).
Plaque composition may predict the evolution of carotid artery stenosis rather than its sole extent. The grey scale median (GSM) value is a reproducible and standardized value to report plaque echogenicity as an indirect measure of its composition. We monitored plaque composition in asymptomatic subcritical carotid stenosis and evaluated the effect of an oral modulating calcification factor (vitamin K2). Carotid plaque composition was assessed by GSM value. Monitoring the effects of standard therapy (acetylsalicylic acid and low-medium dosage statin) (acetylsalicylic acid (ASA) arm) or standard therapy plus vitamins K2 oral supplementation (ASA + K2 arm) over a 12 months period was conducted using an ultrasound scan in a prospective, open-label, randomized controlled trial (PLAK2). Sixty patients on low-medium dosage statin therapy were enrolled and randomized (30 per arm) to either ASA + K2 or ASA alone. Thirty-seven patients (61.6%) showed at 12 months a stable plaque with a mean increase in the GSM value in respect to the baseline of 2.6% with no differences between the two study arms (p = 0.66). Fifteen patients (25%) showed an 8% GSM value reduction respect the baseline with no differences between the two study arms (p = 0.99). At multivariable analysis, the adjusted mean (95% confidence interval) GSM change per month from baseline was greater in the ASA + K2 arm (-0.55 points, p = 0.048) compared to ASA alone (-0.18 points, p = 0.529). Carotid plaque composition monitoring through GSM value represents a laborious procedure. Although its use may not be applied to everyday practice, a specific application consists in evaluating the effect of pharmacological therapy on plaque composition. This 12 months randomized trial showed that the majority of subcritical asymptomatic carotid plaque on treatment with low-medium dosage statin presented a stable or increased echogenicity. Although vitamin K2 beyond standard therapy did not determine a significant change in plaque composition, for those who presented with GSM reduction it did enhance a GSM monthly decline.


[Active form of dihydropteridine reductase in human chorion cells. Possibility of prenatal diagnosis].
Activity of dihydropteridine reductase was studied in human chorion cells for development of accurate methods for prenatal diagnosis of phenylketonuria (especially its most severe form known as lethal hyperphenylalaninemia).


Does replacing intravenous pyelography with noncontrast helical computed tomography benefit patients with suspected acute urolithiasis?
To determine if replacing intravenous pyelography with noncontrast helical computed tomography (NHCT) scanning of the abdomen for the investigation of suspected acute urolithiasis results in shorter stays in the Emergency Department, reduced hospital admissions or fewer interventions. A retrospective review of the charts of all patients who were discharged from the Emergency Department with a diagnosis of acute urolithiasis or renal colic was conducted. Length of stay, hospital admissions and the number of therapeutic interventions were compared for the 5-month period before and the 5-month period after the implementation of NHCT scanning of the abdomen as the primary investigation of suspected acute urolithiasis. Of 230 cases reviewed, 119 met all of the inclusion criteria (61 in the intravenous pyelography group and 58 in the NHCT group). No significant differences were found between the 2 groups on median length of stay in the Emergency Department (7.6 h v. 6.2 h), hospital admission rates or post-test therapeutic interventions. Replacing intravenous pyelography with NHCT scanning for the investigation of suspected acute urolithiasis does not result in significantly shorter stays, reduced hospital admissions or fewer interventions for patients.


Comparison of early season suppressive anthelmintic prophylactic methods for parasitic gastroenteritis and bronchitis in calves.
A comparison was made of the material costs and effectiveness of three methods of early season suppression by anthelmintic medication of Ostertagia species and two of Dictyocaulus viviparus in calves, each method suppressing faecal egg output for different lengths of time from the start of spring grazing. The anthelmintics used were: Morantel bolus administered five days before going to grazing; oxfendazole given three times at three, six and nine weeks after the start of grazing and ivermectin injected three, eight and 13 weeks after going to pasture. The effectiveness of each was estimated by comparison with worm numbers in untreated control calves. Oxfendazole, which was active for the shortest time (about 65 days) from the start of grazing (May 1), produced a 78.1 per cent reduction in Ostertagia species and an 84.4 per cent reduction in D viviparus. The morantel bolus was estimated to be active for 90 days and resulted in a 94.3 per cent reduction of Ostertagia species. The ivermectin treatment, which, because of the prolonged excretion of the chemical and different sensitivity of worm species, was estimated to suppress Ostertagia species for 105 days and D viviparus for 119 days, caused reductions of 98.7 per cent of the former and 97.4 per cent of the latter species. Material costs per calf were estimated at pounds 1.25 for oxfendazole, pounds 2.00 for ivermectin and pounds 10 for the morantel bolus.


Tracking control of a rolling disk.
The tracking control of a disk rolling without slipping on the horizontal (X, Y)-plane is considered. The motion of the disk can be controlled via a tilting torque and a pedaling torque. The concept of path controllability of the disk is introduced and then used to calculate control laws such that the disk tracks a given path in the (X, Y)-plane.


Methylated bismuth, but not bismuth citrate or bismuth glutathione, induces cyto- and genotoxic effects in human cells in vitro.
Bismuth compounds are widely used in industrial processes and products. In medicine, bismuth salts have been applied in combination with antibiotics for the treatment of Helicobacter pylori infections, for the prevention of diarrhea, and in radioimmunotherapy. In the environment, bismuth ions can be biotransformed to the volatile bismuth compound trimethylbismuth (Me3Bi) by methanobacteria. Preliminary in-house studies have indicated that bismuth ions are methylated in the human colon by intestinal microflora following ingestion of bismuth-containing salts. Information concerning cyto- and genotoxicity of these biomethylated products is limited. In the present study, we investigated the cellular uptake of an organic bismuth compound [monomethylbismuth(III), MeBi(III)] and two other bismuth compounds [bismuth citrate (Bi-Cit) and bismuth glutathione (Bi-GS)] in human hepatocytes, lymphocytes, and erythrocytes using ICP-MS. We also analyzed the cyto- and genotoxic effects of these compounds to investigate their toxic potential. Our results show that the methylbismuth compound was better taken up by the cells than Bi-Cit and Bi-GS. All intracellularly detected bismuth compounds were located in the cytosol of the cells. MeBi(III) was best taken up by erythrocytes (36%), followed by lymphocytes (17%) and hepatocytes (0.04%). Erythrocytes and hepatocytes were more susceptible to MeBi(III) exposure than lymphocytes. Cytotoxic effects of MeBi(III) were detectable in erythrocytes at concentrations >4 microM, in hepatocytes at >130 microM, and in lymphocytes at >430 microM after 24 h of exposure. Cytotoxic effects for Bi-Cit and Bi-GS were much lower or not detectable in the used cell lines up to a tested concentration of 500 microM. Exposure of lymphocytes to MeBi(III) (250 microM for 1 h and 25 microM/50 microM for 24 h) resulted in significantly increased frequencies of chromosomal aberrations (CA) and sister chromatid exchanges (SCE), whereas Bi-Cit and Bi-GS induced neither CA nor SCE. Our study also showed an intracellular production of free radicals caused by MeBi(III) in hepatocytes but not in lymphocytes. These data suggest that biomethylation of bismuth ions by the intestinal microflora of the human colon leads to an increase in the toxicity of the primary bismuth salt.


influence of Mg++ and reserpine on the reactivity of rat seminal vesicle to 5-hydroxytryptamine.
The effects of varying the concentration of Mg++ in the incubation medium on the contractile responses of rat isolated seminal vesicle to 5-hydroxytryptamine (5-HT) were investigated in the absence and presence of EDTA. Preparations incubated in Mg++-free and Mg++ excess media showed supersensitivity and subsensitivity to 5-HT respectively. In the presence of EDTA alterations in the sensitivity to 5-HT produced by varying the concentrations of Mg++ were comparatively less. Preparations obtained from reserpinized animals showed subsensitivity in normal and Mg++ excess media and supersensitivity in Mg++-free medium. In the presence of EDTA, reserpinized preparations showed slight supersensitivity in normal Mg++ medium, marked supersensitivity in Mg++-free medium and lesser subsensitivity in Mg++ excess medium. Probably EDTA by more effectively removing Mg++ from the membrane binding sites by chelation makes the membrane permeable to Ca++ leading to supersensitivity to 5-HT (observed only in the presence of EDTA). These results suggest that the failure of reserpine to induce supersensitivity to 5-HT in rat seminal vesicle may be due to an enhanced antagonism of Mg++ on Ca++ movements in this preparation due to the poor capacity of rat tissue to retain Ca++ (Krishnamurty and Grollman, 1976).


Heterodimers of zinc and free-base chlorophyll derivatives co-assembled in biomimetic chlorosomal J-aggregates.
Light-harvesting antennas are one of the most important types of apparatus that use solar energy for natural and artificial photosynthesis. Bacteriochlorophyll (BChl) and chlorophyll (Chl) pigments play key roles in absorbing photons from the sun, migrating/transferring the singlet excitation energy, and transferring electrons. Chlorosomes, the main light-harvesting antennas of photosynthetic green bacteria, contain self-aggregates composed of a large number of BChl-c, d, e, and f molecules without any assistance from peptides, and such J-aggregates transfer the harvested energy to BChl-a pigments of baseplate proteins. In building such green bacterial light-harvesting antennas in artificial systems, self-aggregative model compounds of chlorosomal BChl-c-f pigments and their energy transferring systems have been reported to use synthetic (B)Chl derivatives in organic and aqueous solutions. In this study, we report the co-assembly and energy transfer of chlorin and porphyrin model compounds with covalently linked synthetic zinc and free-base (B)Chl heterodimers having different π-skeletons (bacteriochlorin, chlorin, and porphyrin). The co-assemblies of the self-aggregating BChl-d model having a porphyrin π-skeleton with heterodimers were first studied and showed singlet excitation energy transfer from the photoexcited chlorosomal J-aggregates to a free-base bacteriochlorin or chlorin moiety of the heterodimers. The present co-assembly is a good light-harvesting model system that mimics chlorosomes of photosynthetic green bacteria.


Central neurogenic neuroprotection: central neural systems that protect the brain from hypoxia and ischemia.
The brain can protect itself from ischemia and/or hypoxia by two distinct mechanisms which probably involve two separate systems of neurons in the CNS. One, which mediates a reflexive neurogenic neuroprotection, emanates from oxygen-sensitive sympathoexcitatory reticulospinal neurons of the RVLM. These cells, excited within seconds by reduction in blood flow or oxygen, initiate the systemic vascular components of the oxygen conserving (diving) reflex. They profoundly increase rCBF without changing rCGU and, hence, rapidly and efficiently provide the brain with oxygen. Upon cessation of the stimulus the systemic and cerebrovascular adjustments return to normal. The system mediating reflex protection projects via as-yet-undefined projections from RVLM to upper brainstem and/or thalamus to engage a small population of neurons in the cortex which appear to be dedicated to transducing a neuronal signal into vasodilation. It also appears to relay the central neurogenic vasodilation elicited from other brain regions, including excitation of axons innervating the FN. This mode of protection would be initiated under conditions of global ischemia and/or hypoxemia because the signal is detected by medullary neurons. The second neuroprotective system is represented in intrinsic neurons of the cerebellar FN and mediates a conditioned central neurogenic neuroprotection. The response can be initiated by excitation of intrinsic neurons of the FN and does not appear dependent upon RVLM. The pathways and transmitters that mediate the effect are unknown. The neuroprotection afforded by this network is long-lasting, persisting for almost two weeks, and is associated with reduced excitability of cortical neurons and reduced immunoreactivity of cerebral microvessels. This mode of neuroprotection, moreover, is not restricted to focal ischemia, as we have demonstrated that it also protects the brain against global ischemia and excitotoxic cell death. That the brain may have neuronal systems dedicated to protecting itself from injury, at first appearing to be a novel concept, is, upon reflection, not surprising since the brain is not injured in naturalistic behaviors characterized by very low levels of rCBF, diving and hibernation. An understanding of the pathways, transmitters, and molecules engaged in such protection may provide new insights into novel therapies for a range of disorders characterized by neuronal death.


HHV-8 prevalence, immunosuppression and Kaposi's sarcoma in South Italy.
The identification of HHV-8 has opened the way for numerous epidemiological studies aimed at determining both the prevalence of HHV-8 in various sub-groups of the population (affected or not by KS) and at identifying possible cofactors necessary for the development of KS. We set up a study to evaluate the prevalence of HHV-8 in the South of Italy in KS cases, hospital patients and blood donors and to verify the role of immunosuppression in KS. In KS patients the prevalence of lytic and latent antigens were both 91% (29 positive cases). Lytic and latent antigens have prevalence rates of 20% and 15% respectively in hospital patients. In the donor group the rates were 16% for lytic antigens and 2% for latent antigens. The most recurrent chronic pathology in KS patients was cardiopathy (5 cases). The pathological case histories report 4 cases of Herpes Zoster, 6 of diabetes, one case of hepatitis C who had also had gonorrea. There was also a case, negative to HHV-8, who had had malaria after residing for three years in Oristano in Sardinia (a zone with high endemic malaria). Our study confirms that in Southern Italy there are relatively high prevalences of HHV-8 both in the general population and in blood donors and that immunodysregulation may be involved in the pathogenesis of KS. Other studies are necessary to confirm the sexual transmission of the HHV-8 virus and to better understand the natural history of HHV-8 infection.


Relationship between sporulation medium and germination ability of Mucor racemosus sporangiospores.
Asexual sporangiospores of Mucor racemosus produced on a minimal sporulation medium (M spores) germinated only if glucose, mannose or a complex substrate such as peptone, yeast extract or Casamino acids was present. Once germinated, growth was supported by a wide range of substrates including amino acids, carbohydrates or organic acids. Sporangiospores produced on a nutritionally complex sporulation medium (C-spores) germinated on a wide range of carbon sources. C-spore phenotype was pleiotropic in that sporangiospores capable of germinating on cellobiose could always germinate on glutamate or xylose; but C-spores capable of germinating on xylose or glutamate did not always germinate on cellobiose. There was a hierarchy of substrates capable of initiating germination with glucose = mannose greater than xylose greater than glutamate greater than cellobiose. C-spores also differed from M-spores by initiating germination in the presence of the non-metabolizable glucose analogue 3-O-methylglucose. These results suggest that at least two sporangiospore phenotypes are produced depending upon the concentration and type of ingredients present in the sporulation medium.


Optical pattern processing utilizing nematic liquid crystals.
This paper describes a new type of optical processing based on the dynamic scattering effect in liquid crystals. It is possible to realize a binary spatial filter with nematic liquid crystals and to control the optical modes by voltage signals. A two-dimensional Walsh function is used for extracting properties of the input patterns. The realization of an 8 x 8 Walsh function filter with nematic liquid crystals is demonstrated. The contrast ratio is over 30 to 1. The variation in transparency is within +/-3%. Experimental results show that it is possible to extract the properties of handwritten characters by detecting the intensity of light transmitted through both the input pattern and the liquid crystal filter.


Integrated therapeutic approach to giant solitary fibrous tumor of the pleura: report of a case and review of the literature.
The fibrous tumors of the pleura are rare primary tumors, accounting for 5% of malignant pleural neoplasms, which generally originate from sub-mesothelial mesenchymal tissue of the visceral pleura. These tumours generally exhibit clinical benign behavior although 12% of solitary fibrous tumors can be malignant and have worse outcomes. These tumors are considered "giant" when the lesion > 15 cm. Surgical treatment is the best choice for both benign and malignant neoplasms. We retrospectively analyzed the main case series of giant fibrous tumors of the pleura. In addition we report our experience of a 76-year-old woman treated by pre-surgical embolization involving implantation of vascular plugs. Surgery was successfully carried out without complications; imaging and functional assessment 6 months post intervention demonstrated both the absence of recurrence and improvement of lung function parameters.


Assessing early-onset hallucinations in the touch-screen generation.
The increasing development of apps for digital devices provides an opportunity for new instruments to assess hallucinations in young individuals. Here we present the Multisensory HAllucinations Scale for Children (MHASC), dedicated to assessing complex early-onset hallucinations. The MHASC will soon be translated into multilanguage versions with the support of the International Consortium of Hallucination Research.


Low-Level MHC Class II Expression Leads to Suboptimal Th Cell Response, Increased Autoaggression, and Heightened Cytokine Inducibility.
The development and activation of MHC class II (MHC-II)-restricted CD4+ T cells are distinct immunological processes that are strictly MHC-II-dependent. To address their relative dependence on MHC-II, we established a novel ENU-induced mutant mouse on the C57BL/6 background, named I-A12%, with ∼8-fold reduced I-A expression on the surface of B cells, dendritic cells, cortical thymic epithelial cells, and medullary thymic epithelial cells. I-A100% and I-A12% mice are highly similar with respect to the numbers of double-positive thymocytes, CD4+CD8- T cells, regulatory T cells, CD4+ T cell marker expression, lifespan, and Th/regulatory T cell function. Despite the demonstration of functional intrathymic negative selection in I-A12% mice, transfer of I-A12% CD25-CD4+ T cells into RAG-knockout hosts revealed increased autoaggression activity against the liver. Compared to I-A100% mice, infection of I-A12% mice with graded doses of Listeria monotcytogenes or influenza virus revealed comparable and significantly reduced generation of Ag-specific CD4+ T cells at high and low infection doses, respectively. A significantly weakened Ag-specific recall cytokine production response was also found for I-A12% mice previously infected with a relative low dose of L. monocytogenes CD44hiCD4+ T cells from I-A100% and I-A12% mice previously infected with a relatively high L. monocytogenes dose displayed highly similar Ag-specific multicytokine production profiles. In contrast, polyclonal activation of endogenous memory-like I-A12% CD44hiCD4+ T cells revealed highly elevated production of multiple cytokines. Our results demonstrate that there exist distinct thresholds for different MHC-II-dependent immunological processes. The I-A12% mutant mouse model we describe in the present study is a valuable tool for investigations on the quantitative cause-effect relationship in MHC-II-dependent normal and autoimmune responses.


Consuming sunflower pollen reduced pathogen infection but did not alter measures of immunity in bumblebees.
Certain diets can benefit bee health by reducing pathogens, but the mechanism(s) driving these medicinal effects are largely unexplored. Recent research found that sunflower (Helianthus annuus) pollen reduces the gut pathogen Crithidia bombi in the common eastern bumblebee (Bombus impatiens). Here, we tested the effects of sunflower pollen and infection on two bee immune metrics to determine whether sunflower pollen diet drives changes in host immunity that can explain this medicinal effect. Bees were infected with C. bombi or not and given either sunflower or wildflower pollen. Subsequently, bees received a benign immune challenge or were left naive to test the induced and constitutive immune responses, respectively. We measured haemolymph phenoloxidase activity, involved in the melanization cascade, and antibacterial activity. Sunflower pollen reduced C. bombi infection, but we found no significant pollen diet effect on either immune measure. Phenoloxidase activity was also not affected by C. bombi infection status; however, uninfected bees were more likely to have measurable constitutive antibacterial activity, while infected bees had higher induced antibacterial activity. Overall, we found that sunflower pollen does not significantly affect the immune responses we measured, suggesting that the mechanisms underlying its medicinal effect do not involve these bee immune parameters. This article is part of the theme issue 'Natural processes influencing pollinator health: from chemistry to landscapes'.


[Acute appendicitis in children with atypical location of the vermiform process].
The analysis of case records of 19,966 children operated on for acute appendicitis at the Belorussian centre of pediatric surgery for 15 years (from 1970 till 1984) is presented. In 2796 (14%) children, the atypical location of the vermiform process was diagnosed at operation. In atypical location of the vermiform process, the clinical manifestations of acute appendicitis differ from those in its common location: abdominal pain is most frequently (47.8% of cases) localized outside the inguinal region, less often, tensity of the anterior abdominal wall muscles, symptoms of peritoneal irritation are noted. Children with acute appendicitis in atypical location of the vermiform process twice later (when compared with patients with its typical location) are admitted to the hospital, and as well twice longer are observed before the operation. The incidence of complications after the operation is more than two-fold greater.


Deficient approaches to human neuroimaging.
Functional magnetic resonance imaging (fMRI) is the workhorse of imaging-based human cognitive neuroscience. The use of fMRI is ever-increasing; within the last 4 years more fMRI studies have been published than in the previous 17 years. This large body of research has mainly focused on the functional localization of condition- or stimulus-dependent changes in the blood-oxygenation-level dependent signal. In recent years, however, many aspects of the commonly practiced analysis frameworks and methodologies have been critically reassessed. Here we summarize these critiques, providing an overview of the major conceptual and practical deficiencies in widely used brain-mapping approaches, and exemplify some of these issues by the use of imaging data and simulations. In particular, we discuss the inherent pitfalls and shortcomings of methodologies for statistical parametric mapping. Our critique emphasizes recent reports of excessively high numbers of both false positive and false negative findings in fMRI brain mapping. We outline our view regarding the broader scientific implications of these methodological considerations and briefly discuss possible solutions.


An efficient chronic unpredictable stress protocol to induce stress-related responses in C57BL/6 mice.
Exposure to chronic stress can have broad effects on health ranging from increased predisposition for neuropsychiatric disorders to deregulation of immune responses. The chronic unpredictable stress (CUS) protocol has been widely used to study the impact of stress exposure in several animal models and consists in the random, intermittent, and unpredictable exposure to a variety of stressors during several weeks. CUS has consistently been shown to induce behavioral and immunological alterations typical of the chronic stress-response. Unfortunately C57BL/6 mice, one of the most widely used mouse strains, due to the great variety of genetically modified lines, seem to be resistant to the commonly used 4-week-long CUS protocol. The definition of an alternative CUS protocol allowing the use of C57BL/6 mice in chronic stress experiments is a need. Here, we show that by extending the CUS protocol to 8 weeks is possible to induce a chronic stress-response in C57BL/6 mice, as revealed by abrogated body weight gain, increased adrenals weight, and an overactive hypothalamic-pituitary-adrenal axis with increased levels of serum corticosterone. Moreover, we also observed stress-associated behavioral alterations, including the potentiation of anxious-like and depressive-like behaviors and a reduction of exploratory behavior, as well as subtle stress-related changes in the cell population of the thymus and of the spleen. The present protocol for C57BL/6 mice consistently triggers the spectrum of CUS-induced changes observed in rats and, thus, will be highly useful to researchers that need to use this particular mouse strain as an animal model of neuropsychiatric disorders and/or immune deregulation related to CUS.


Recreational fishing, health and well-being: findings from a cross-sectional survey.
We evaluated the associations between marine recreational fishing, stress, seafood consumption, and sleep quality in a cross-sectional questionnaire-based survey of a convenience sample of 244 fishers recruited in 2019 in Spain. Fishers' stress levels were moderate, with a mean stress index score of 36.4 units on a scale from 14 (very low stress) to 70 (very high). Their average emotional condition was positive, with a mean index of negative affect of 7.8 units on a scale from 5 (very low negative affect) to 25 (very high). Seafood intake was low, with a mean index of seafood in diets of 38.0 units on a scale from 20 (very low seafood consumption) to 160 (very high). Fishers' perceived quality of night sleep was good because the mean index of sleep problems was 39.5 units on a scale from 21 (very low sleep problems) to 107 (very high). Each hour of self-reported monthly fishing activity was associated with 0.016 units of lower stress score. Thus, the most engaged fishers reported up to 15.4% lower stress score than less avid fishers. Since recreational fishing is a highly accessible outdoor activity for people in older age groups, it is possible that public health could be improved by access to sustainably managed recreational fisheries. Fishing engagement was positively associated with seafood intake. Each hour of fishing per month was associated with one-unit higher seafood consumption. The higher seafood consumption observed among avid recreational fishers compared with less avid fishers might have health implications.


Provider-Level Barriers to Human Papillomavirus Vaccination in Survivors of Childhood and Young Adult Cancers.
Purpose: We sought to understand clinician-level barriers to providing HPV vaccination to survivors of childhood and young adult cancers (CYACs). Methods: We conducted 30-minute qualitative interviews with primary care and specialty clinicians who care for survivors of CYACs at our academic medical center. Blinded reviewers analyzed transcripts and used an inductive approach to identify barriers to vaccination in this population. Results: We conducted 24 interviews (n = 11 primary care clinicians, n = 13 oncology clinicians). Thematic analysis revealed that primary care clinicians are universally viewed as holding ultimate responsibility for human papillomavirus (HPV) vaccination among survivors of CYACs. Both primary care and oncology clinicians believed vague, inconsistent HPV guidelines engendered uncertainty toward HPV vaccination's role and timing following completion of CYAC therapies. As such, compared with other vaccines, the HPV vaccination is not as consistently offered to survivors. Respondents identified direct guidance from oncologists to primary care clinicians and to patients as a potential strategy for improving HPV vaccination rates in this population. Finally, oncology clinicians frequently deprioritize the issue of preventing second, noniatrogenic cancers and consequently miss opportunities to discuss vaccination's merits with their patients. Conclusions: Despite not holding ultimate responsibility for vaccination, oncology clinicians have an opportunity to play an important role in ensuring access and overcoming hesitancy among survivors of CYACs. Developing clearer and more collaborative guidelines, helping to integrate vaccination into institutional electronic health record protocols, offering direct guidance to primary care colleagues, and participating in conversations with survivors of CYACs may help improve vaccination rates.


C9orf72 BAC Mouse Model with Motor Deficits and Neurodegenerative Features of ALS/FTD.
To define how the C9orf72 GGGGCC expansion mutation causes ALS/FTD and to facilitate therapy development, a mouse model that recapitulates the molecular and phenotypic features of the disease is urgently needed. Two groups recently reported BAC mouse models that produce RNA foci and RAN proteins but, surprisingly, do not develop the neurodegenerative or behavioral features of ALS/FTD. We now report a BAC mouse model of C9orf72 ALS/FTD that shows decreased survival, paralysis, muscle denervation, motor neuron loss, anxiety-like behavior, and cortical and hippocampal neurodegeneration. These mice express C9orf72 sense transcripts and upregulated antisense transcripts. In contrast to sense RNA foci, antisense foci preferentially accumulate in ALS/FTD-vulnerable cell populations. RAN protein accumulation increases with age and disease, and TDP-43 inclusions are found in degenerating brain regions in end-stage animals. The ALS/FTD phenotypes in our mice provide a unique tool that will facilitate developing therapies targeting pathways that prevent neurodegeneration and increase survival.


Clinical manifestation of norovirus gastroenteritis in health care settings.
Noroviruses are generally believed to cause relatively mild gastroenteritis of short duration in otherwise healthy adults. However, outbreaks in health care settings are common and affect vulnerable populations. During 2002-2003, a total of 4 major hospitals, 11 community hospitals, and 135 nursing homes in the county of Avon, England, were prospectively monitored for outbreaks of gastroenteritis. For 482 hospital staff, 166 nursing home staff, and 266 nursing home residents, the median duration of norovirus gastroenteritis was 2 days, with 75% achieving complete recovery within 3 days. The median duration of norovirus gastroenteritis for 730 hospital patients was 3 days (75% of the patients achieved complete recovery within 5 days), which was significantly longer than that for all other groups (P<.001). Therefore, infection in hospitalized persons may be more severe than that in other groups in the community at large. This increased duration of acute illness should be considered when implementing measures to prevent transmission in hospital settings.


In-silico homology assisted identification of inhibitor of RNA binding against 2019-nCoV N-protein (N terminal domain).
The N terminal domain (NTD) of Nucleocapsid protein (N protein) of coronavirus (CoV) binds to the viral (+) sense RNA and results in CoV ribonucleoprotien (CoV RNP) complex, essential for the virus replication. In this study, the RNA-binding N terminal domain (NTD) of the N protein was targeted for the identification of possible inhibitors of RNA binding. Two NTD structures of N proteins were selected (2OFZ and 1SSK, 92% homology) for virtual screening of 56,079 compounds from Asinex and Maybridge library to identify top 15 hits for each of the targets based on 'docking score'. These top-hits were further screened for MM-GBSA binding free energy, pharmacokinetic properties (QikProp) and drug-likeness (SwissADME) and subjected to molecular dynamics (MD) studies. Two suitable binders (ZINC00003118440 and ZINC0000146942) against the target 2OFZ were identified. ZINC00003118440 is a theophylline derivative under the drug class 'bronchodilators' and further screening with approved bronchodilators was also studied to identify their ability to bind to the RNA binding region on the N protein. The other identified top hit is ZINC0000146942, which is a 3,4dihydropyrimidone class molecule. Hence this study suggests two important class of compounds, theophylline and pyrimidone derivaties as possible inhibitors of RNA binding to the N terminal domain of N protein of coronavirus, thus opening new avenues for in vitro validations. Communicated by Ramaswamy H. Sarma.


Privacy-preserving techniques for decentralized and secure machine learning in drug discovery.
Data availability, data security, and privacy concerns often hamper optimal performance efficiency of machine learning (ML) techniques. Therefore, novel techniques for the utilization of private/sensitive data in the field of drug discovery have been proposed for ML model-building tasks. Some examples of the different techniques are secure multiparty computation, distributed deep learning, homomorphic encryption, blockchain-based peer-to-peer networking, differential privacy, and federated learning, as well as combinations of such techniques. In this paper, we present an overview of these techniques for decentralized ML to illustrate its benefits and drawbacks in the field of drug discovery.


Spontaneous bacterial peritonitis in Wilson's disease.
Spontaneous bacterial peritonitis associated with Wilson's disease has been previously reported in only one case. We report two cases where this infection developed and seriously complicated the course of illness. As with spontaneous bacterial peritonitis occurring with other underlying diseases, early diagnosis and treatment are critical to improved outcome. Recognition that it can occur with Wilson's disease is important, and paracentesis should be performed without delay when suspicious features are present.


On the peculiarities of CBED pattern formation revealed by multislice simulation.
A modified multislice method has been developed for calculations of Convergent Beam Electron Diffraction (CBED) patterns. The validity of the method for HOLZ- and Kikuchi-line calculations has been proofed by comparison to Bloch-wave calculations. The application of the method leads to the new understanding of CBED patterns formation. Dynamical scattering of weak HOLZ reflections plays the key role in the appearance of deficient lines in the central CBED disk. Different HOLZ lines do have significantly different and extended scattering areas; the central 000 CBED disk, consequently, contains structural information from an area around the primary beam which is determined by the Bragg angle of HOLZ reflections and the thickness of the sample. A variation of lattice parameters, if present within this area, results in artificial symmetry violations of the pattern and in changes of line profiles.


Coupling between physiological TSPO expression in brain and myocardium allows stabilization of late-phase cerebral [18F]GE180 PET quantification.
PET imaging of the 18 kDa translocator protein (TSPO), a biomarker of microglial activity, receives growing interest in clinical and preclinical applications of neuroinflammatory and neurodegenerative brain diseases. In globally affected brains, intra-cerebral pseudo reference regions are not feasible. Consequently, many brain-independent approaches have been attempted, including SUV analysis and normalization to muscle- or heart uptake, aiming to stabilize quantitative analysis. In this study, we systematically compared different image normalization methods for static late phase TSPO-PET imaging of rodent brain. We first obtained gamma counter measurements for gold standard quantitation of [18F]GE180 uptake in brain of C57Bl/6 mice (N = 10) after PET, aiming to identify factors contributing significantly to the quantitative results. Subsequently, data from a large cohort of C57Bl/6 mice (N = 79) were compiled to precisely determine the weighted influence and variance attributable these factors by regression analysis. Scan-rescan variability and agreement with histology were used to validate the tested normalization methods in an Alzheimer's disease (AD) mouse model with pathologically increased TSPO expression (PS2APP; N = 24). Longitudinal data from AD model mice (N = 10) scanned at four different ages were used to challenge and validate the different normalization methods in a practical application. Gamma counter results revealed that injected dose, body weight and PET-measured radioactivity concentration in the ventral myocardium all significantly accounted for [18F]GE180 activity in the brain. Skeletal muscle activity had high test-retest variance in this PET only application and was therefore pursued no further. Regression analysis of the large scale evaluation showed that scaling to injected dose or SUV analysis accounted for little variance in brain activity (R2 < 0.5), but inclusion of myocardial activity together with injected dose and body weight in the regression model accounted for most of the variance in brain uptake (R2 = 0.94). Scan-rescan stability, correlation with histology and applicability for longitudinal examination in the disease model were also significantly improved by inclusion of myocadial uptake in the quantitative model. Cerebral and myocardial TSPO expression are highly coupled under physiological conditions. Myocardial uptake has great potential for stabilization of static late phase [18F]GE180 quantification in brain in the absence of a valid intra-cerebral pseudo-reference region.


Syphilitic proctitis mimicking rectal cancer: A case report.
Syphilitic proctitis is a rare disease. It usually presents as proctitis, ulcer and neoplasm but lacks pathognomonic clinical symptoms. It is, therefore, difficult to diagnose and is occasionally treated inappropriately. We report the case of a 51-year-old man who had a hard, ulcerated mass, which occupied the circumference of the rectal wall and which mimicked a rectal tumor. Fortunately, positive finding from routine toluidine red unheated serum test and treponema pallidum particle agglutination tests made us reevaluate the patient and led us to suspect syphilitic proctitis. This diagnosis was finally confirmed after successful penicillin G benzathine therapy which made surgery unnecessary.


Wisdom of patients: predicting the quality of care using aggregated patient feedback.
The Care Quality Commission (CQC) is responsible for ensuring the quality of healthcare in England. To that end, CQC has developed statistical surveillance tools that periodically aggregate large numbers of quantitative performance measures to identify risks to the quality of care and prioritise its limited inspection resource. These tools have, however, failed to successfully identify poor-quality providers. Facing continued budget cuts, CQC is now further reliant on an 'intelligence-driven', risk-based approach to prioritising inspections and a new effective tool is required. To determine whether the near real-time, automated collection and aggregation of multiple sources of patient feedback can provide a collective judgement that effectively identifies risks to the quality of care, and hence can be used to help prioritise inspections. Our Patient Voice Tracking System combines patient feedback from NHS Choices, Patient Opinion, Facebook and Twitter to form a near real-time collective judgement score for acute hospitals and trusts on any given date. The predictive ability of the collective judgement score is evaluated through a logistic regression analysis of the relationship between the collective judgement score on the start date of 456 hospital and trust-level inspections, and the subsequent inspection outcomes. Aggregating patient feedback increases the volume and diversity of patient-centred insights into the quality of care. There is a positive association between the resulting collective judgement score and subsequent inspection outcomes (OR for being rated 'Inadequate' compared with 'Requires improvement' 0.35 (95% CI 0.16 to 0.76), Requires improvement/Good OR 0.23 (95% CI 0.12 to 0.44), and Good/Outstanding OR 0.13 (95% CI 0.02 to 0.84), with p<0.05 for all). The collective judgement score can successfully identify a high-risk group of organisations for inspection, is available in near real time and is available at a more granular level than the majority of existing data sets. The collective judgement score could therefore be used to help prioritise inspections.


Psychosocial and behavioral functioning and their relationship to seizure timing in children with benign epilepsy with centrotemporal spikes.
Psychosocial and behavioral problems have been reported in children with benign epilepsy with centrotemporal spikes (BECTS). Distinctive features of typical BECTS associated with cognitive and behavioral problems have not clearly been defined. We aimed to identify psychosocial and behavioral functioning and their relationship to seizure timing in BECTS. Consecutive patients with BECTS were recruited from the pediatric neurology outpatient clinic between May 2015 and May 2016. The patients were divided into two subgroups in according to seizure timing; group 1 consisted of patients with seizures only in the morning short before awakening, and group 2 consisted of patients with seizure shortly after falling asleep or in both time periods. Neuropsychological and behavioral evaluation in patients and healthy controls were examined using the Wechsler Intelligence Scale for Children-Revised test and the Turkish version of Strengths and Difficulties Questionnaire (SDQ). The participants comprised 46 children with BECTS and 49 healthy controls aged 7-16years. There was no significant difference between group 1, group 2, and control group regarding intelligence quantity in full-scale or verbal and performance subscales. Behavioral scores for overall stress significantly differed between group 2 and controls on the SDQ test, while group 1 and control group had no difference on the SDQ scores. Patients with BECTS who have seizure shortly after falling asleep may have a tendency towards behavior difficulties.


Benchmarking a Fast Proton Titration Scheme in Implicit Solvent for Biomolecular Simulations.
pH is a key parameter for technological and biological processes, intimately related to biomolecular charge. As such, it controls biomolecular conformation and intermolecular interactions, for example, protein/RNA stability and folding, enzyme activity, regulation through conformational switches, protein-polyelectrolyte association, and protein-RNA interactions. pH also plays an important role in technological systems in food, brewing, pharma, bioseparations, and biomaterials in general. Predicting the structure of large proteins and complexes remains a great challenge experimentally, industrially, and theoretically, despite the variety of numerical schemes available ranging from Poisson-Boltzmann approaches to explicit solvent based methods. In this work we benchmark a fast proton titration scheme against experiment and several theoretical methods on the following set of representative proteins: [HP36, BBL, HEWL (triclinic and orthorhombic), RNase, SNASE (V66K/WT, V66K/PHS, V66K/Δ+PHS, L38D/Δ+PHS, L38E/Δ+PHS, L38K/Δ+PHS), ALAC, and OMTKY3]; routinely used in similar tests due to the diversity of their structural features. Our scheme is rooted in the classical Tanford-Kirkwood model of impenetrable spheres, where salt is treated at the Debye-Hückel level. Treating salt implicitly dramatically reduces the computation time, thereby circumventing sampling difficulties faced by other numerical schemes. In comparison with experimental measurements, our calculated pKa values have the average, maximum absolute, and root-mean-square deviations of 0.4-0.9, 1.0-5.2, and 0.5-1.2 pH units, respectively. These values are within the ranges commonly observed in theoretical models. They are also in the large majority of the cases studied here more accurate than the NULL model. For BBL, ALAC, and OMTKY3, the predicted pKa are closer to experimental results than other analyzed theoretical data. Despite the intrinsic approximations of the fast titration scheme, its robustness and ability to properly describe the main system physics is confirmed.


HOMER3 promotes non-small cell lung cancer growth and metastasis primarily through GABPB1-mediated mitochondrial metabolism.
Cancer metabolism has emerged as a major target for cancer therapy, while the state of mitochondrial drugs has remained largely unexplored, partly due to an inadequate understanding of various mitochondrial functions in tumor contexts. Here, we report that HOMER3 is highly expressed in non-small cell lung cancer (NSCLC) and is closely correlated with poor prognosis. Lung cancer cells with low levels of HOMER3 are found to show significant mitochondrial dysfunction, thereby suppressing their proliferation and metastasis in vivo and in vitro. At the mechanistic level, we demonstrate that HOMER3 and platelet-activating factor acetylhydrolase 1b catalytic subunit 3 cooperate to upregulate the level of GA-binding protein subunit beta-1 (GABPB1), a key transcription factor involved in mitochondrial biogenesis, to control mitochondrial inner membrane genes and mitochondrial function. Concurrently, low levels of HOMER3 and its downstream target GABPB1 led to mitochondrial dysfunction and decreased proliferation and invasive activity of lung cancer cells, which raises the possibility that targeting mitochondrial synthesis is an important and promising therapeutic approach for NSCLC.


Youth's Daily Activities and Situational Triggers of Gunshot Assault in Urban Environments.
Although previous research has made progress in identifying individuals predicted to face an elevated risk of being shot, it is not clear how that risk varies within individuals based on the contexts they encounter as they navigate daily life. The current study examines how the convergence of individual risk activity and neighborhood disadvantage and disorder triggers the risk of being shot. Using a novel geographic information system application, 123 male gunshot assault victims between 10 and 24 years old in Philadelphia, Pennsylvania, described their minute-by-minute movements over the course of the day of the gunshot assault. Through latent class analysis, the primary exposure was real-life circumstance where nine theoretically informed risk factors converged, compared with two other circumstances. Case-crossover analyses of subjects' 10-minute segments of full-day activities compared gunshot assault victims at the time of assault with themselves earlier in the day. Compared to when individuals were exposed to minimal situational risk or were mainly exposed to neighborhood disadvantage and disorder, the concurrence of risk activity and neighborhood disadvantage and disorder was associated with a 9.90 (95% CI: 2.72-36.14) and 6.06 (95% CI: 2.78-13.22) times higher risk of being shot. Importantly, the likelihood of being in the high-risk circumstance increased systematically over the course of the day leading up to the time when young individuals were shot. After controlled individual's propensity to be shot (e.g., inherent traits), the concurrence of situational risks emerged as significant triggers of gunshot assault. The findings suggest potential for community-based gunshot violence interventions.


Residents' Concerns Regarding Schools Designated as Evacuation Shelters.
The purpose of this study was to investigate the perceptions of residents who are knowledgeable about the schools used as shelters. The target group was comprised of teachers and guardians of children enrolled in 4 schools, which were selected in the vicinity of District B, which was severely damaged by heavy rain in 2014. A qualitative descriptive study design was used. The number of surveys collected was 1702 (collection rate 62.2%). A total of 1017 clauses were entered, and 85 codes were generated for 7 categories. The guardians and teachers believed that there was a discrepancy between shelter capacity and the real situation; citing a lack of information, and ill preparedness of the schools for disasters as some of the problems. Based on the knowledge and experiences from previous disasters, they responded that there was inadequate management, and evacuees faced difficulty living comfortably in the shelters. In order to use school facilities during any disaster, it is important for residents to recognize the need to solve problems, and to work with local governments to support improvements. This realization reveals a sense of ownership of emergency shelters and prevents confusion among residents. Also, this will help people to prepare for disasters and prevent disruption during evacuation.


Disaggregating Mass Public Shootings: A Comparative Analysis of Disgruntled Employee, School, Ideologically motivated, and Rampage Shooters.
This study compares the demographic, background, motivation, and pre-event and event-level behaviors across four types of mass public shooters: disgruntled employee, school, ideologically motivated, and rampage offenders. Using a database containing detailed information on 318 mass public shootings that occurred in the United States between 1966 and 2017, we find systematic differences in the characteristics, motivations, target selection, planning, and incident-level behaviors among these offenders. The results show that ideologically motivated shooters to be the most patient, and methodical, and as a result the most lethal. Conversely, disgruntled employees, who are driven by revenge, tend to have little time to plan and consequently are the least lethal shooters. These, among other differences, underscore the need for prevention strategies and policies to be tailored to specific types of offenders. Furthermore, the results also highlight commonalities across offender type, suggesting that the social and psychological pathways to violence are universal across offenders.


A new method for the measurement of protein turnover.
A new technique for the determination of rate constants of protein degradation is described. By using the method, half-lives of total soluble protein of Lemna minor during growth on full culture medium and distilled water were measured. The method involves incubating Lemna on a growth medium containing 3H2O. After a short exposure (20 min) to 3H-labelled culture medium, 3H was found in soluble amino acids, especially aspartate, glutamate, glutamine and alanine. After transfer to a 3H-free medium for 30 min, 80% of the 3H originally present in soluble amino acids was lost. These results suggest that 3H enters and leaves amino acids at the alpha-carbon atom, a conclusion supported by the observed labelling of glutamates. The exchange of H and 3H on the alpha-carbon atom is catalysed by transaminases and the speed of this exchange ensures that when the 3H2O is removed, the 3H in free amino acids is rapidly lost, thereby eliminating problems connected with metabolic pools and recycling. After an exposure of 20 min to 3H-labelled medium all protein amino acids, except for arginine, were found to be radioactive. The loss of radioactivity from protein amino acids was used to measure protein degradation.


Comparison of Dorsal and Buccal Approaches for Surgical Extraction of the Mandibular Canine Tooth in Cat Specimens Using Radiographic and Computed Tomographic Analysis.
The purpose of this study was to compare the buccal and dorsal approaches for surgical extraction of mandibular canine teeth in the cat. Parameters evaluated included extraction time, surgical wound closure time, volume of bone loss, and incidence of complications. Both mandibular canine teeth were extracted from each of 12 feline cadaver specimens, using a different surgical approach for each side, and recording the time for extraction and closure. Computed tomography was used to measure the volume of each rostral mandible before and after extraction, and the bone lossfor each approach was computed. The mean extraction time and volume of bone loss was less for the buccal approach, but the results were not statistically significant (P = 0.22 and P = 0.32, respectively). The closure time for the dorsal approach was significantly less than for the buccal approach (P = 0.023). Total time (extraction plus closure time) was not significantly different between the approaches (P = 0.40). There were no complications associated with either approach.


Diagnosis and management of Parkinson's disease dementia.
Parkinson's disease (PD) has long been considered predominantly a motor disorder. However, its frequent association with dementia, which contributes significantly to the morbidity and mortality of the condition, is gaining increasing recognition. PD dementia (PDD) has a unique clinical profile and neuropathology, distinct from Alzheimer's disease (AD). Cholinergic deficits, a feature of both AD and PDD, underlie the rationale for cholinesterase inhibitor therapy in both conditions. In clinical practice, it is important that PDD should be recognised and appropriately treated. This review aims to outline the recently proposed clinical diagnostic criteria for PDD and to summarise the guidelines/recommendations published since 2006 on the use of cholinesterase inhibitors in the management of PDD. Although the cholinesterase inhibitor rivastigmine has recently been approved for the management of PDD, there remains a need for the development of novel therapies that can affect key mechanisms of the disease or prevent/delay patients with PD and mild cognitive impairment from progressing to PDD.


Primary dural lymphoma: a novel concept of heterogeneous disease.
Spinal primary dural lymphoma (PDL) is uncommon with a total of 37 previous well-documented cases reported, including one diagnosed in the authors' institution. More recently we encountered an additional case of spinal PDL that, similarly to our previous case, was grade 1-2 follicular B-cell PDL. Our two cases were diagnosed over a 3-year interval in a 72-year-old female and a 74-year-old male, respectively. An exhaustive literature review on PDL was performed consequently to reveal that: (i) spinal and cerebral sites of involvement by PDL are constantly mutually exclusive; and (ii) unlike cerebral PDL, which is usually of marginal zone B-cell type, only two of the 38 cases of spinal PDL were diagnosed as such, diffuse large B-cell lymphoma being the most commonly encountered type in the spine. This divergence infers that, in contrast to the prevailing concept that PDL is a unique disease group, PDL appears to be rather heterogeneous with a difference in predilection of lymphoma type for the anatomical site of dural involvement. Such a site-specific lymphoma-type predilection phenomenon, well-recognized in other organ systems, has not been acknowledged in PDL. This report brings new insights into PDL, and may contribute to a better understanding of nervous system pathophysiology and lymphoma classification.


[Effect of different types of excessive feeding on the state of vascular walls in rats].
Immature male rats were overfed for a long time with an excess of fats or carbohydrates; this caused obesity. Carbohydrate overfeeding led to a greater body weight gain and a greater increase in the weight of epididymal fat and of the adipose cell volume than fat overfeeding, but failed to decrease the lipolytic activity of the aorta. Prolonged fat overfeeding decreased the lipolytic activity of the aorta considerably. Thus, the results of the mentioned experiments showed that the predisposition of the aortic wall to atherogenesis did not correlate with the gain in weight and depended on the character of nutrition.


Constructing the suitable initial configuration of the membrane-protein system in molecular dynamics simulations.
A method for constructing the suitable initial configuration of the membrane-protein system for molecular dynamics (MD) simulations is presented. This method could provide some hydrated initial configurations and help us to determine the best surface area of the system by contracting the surface area and comparing the optimized lowest energy of the system by energy minimization. The gramicidin A (GA) channel in;the fully hydrated dimyristoylphosphatidylcholine (DMPC) bilayer was used as our model. Three configurations with different surface areas were selected and applied for one 400 ps and two 300 ps MD simulations at constant pressure and temperature. All simulations were fairly stable without any constraints. Through analysis of the MD trajectories we found that the system with the best surface area was more stable than the other two systems, whose sizes were changed in the simulations. Further analysis of the bilayer normal length and the order parameters of the lipid alkyl tails indicates that the system with the best surface area shows some characteristics of the L(alpha) phase, while both the smaller and the larger size systems have distinct deviations from the L(alpha) phase that we expect. This illustrates that the correct surface area and the suitable initial configuration have an important influence on the phase of the membrane in the MD simulation. In addition, by comparing the root mean square differences of GA relative to the initial structure and interaction energy between different components of the system for all three systems, we find that the state of the DMPC bilayer has exerted a significant influence on the structure of GA. All these results demonstrate the validity of our method for constructing the initial configuration of the membrane-protein system for MD simulations.


Variation in indicators of respiratory functions among Warsaw adolescents in relation to ambient air pollution and smoking.
This work presents results of studies on secondary school adolescents inhabiting two regions of Warsaw: Sródmiescie district (Downtown) and Miedzylesie (Wawer district - Vicinity), conducted in 2006. These two districts significantly differ according to air pollution, which is much higher in Downtown than in Vicinity. The sample consists of 219 boys and 225 girls aged 13-16 years. The measurements included body height and weight and 8 respiratory variables of lung function: VC, FEV1, TV, MV, IRV, ERV, Ap, RR. Additionally information about education of parents, number of children in family, number of rooms in a house/apartment, smoking habits of pupils and their parents were provided. Sib-sib size, parents education level and number of rooms in apartment or house were included in a principal component analysis (PCA) to obtain a common factor representing general socio-economic status (SES) for families. First factor scores were used as covariates in the further analysis. The influence of air pollution on respiratory variables was evaluated using the analysis of covariance (ANCOVA). The results show that there are significant relations of air pollution to Ap, MV and IRV and smoking habits of individuals on their MV and TV. In the heavier polluted region there are observed longer time of Ap and higher values of IRV There is an opposite reaction of MV on air pollution and smoking. The MV values are lower in the heavier polluted area but presents greater values in smoking individuals. Also values of TV are greater if individuals smoke comparing with non smokers.


Isolation and expression in Escherichia coli of a cDNA clone encoding human beta-glucuronidase.
Mucopolysaccharidosis type VII is a lysosomal storage disease resulting from a deficiency of beta-glucuronidase (BG) activity. To facilitate the investigation of mutation in the disease and provide molecular diagnostic tools for affected families, we have isolated human BG cDNA clones. The SV40-transformed human fibroblast cDNA library of Okayama and Berg [Mol. Cell. Biol. 3 (1982) 280-289] was screened with a fragment of a murine BG cDNA clone (pGUS-1). The 17 human cDNA clones (pHUG) isolated were identical by restriction mapping, varying only in length. The pHUG clones show 80% DNA sequence homology with pGUS-1 in a 198-bp PvuII-SstI restriction fragment. Both pGUS-1 and the pHUG clones contained an open reading frame (ORF) throughout the sequenced region with a predicted amino acid sequence homology of 73%. Expression in Escherichia coli of a 1150-bp fragment of pHUG-1 subcloned in pUC9 resulted in an isopropyl-thio-beta-galactoside (IPTG)-inducible 35-kDal fusion protein which was specifically immunoprecipitated by goat anti-human BG immunoglobulin G (IgG). This evidence provides direct confirmation that the pHUG cDNA clones correspond to human BG.


Modern theories of metabolic control and their applications (review).
Existing, qualitative notions with respect to the way in which enzyme properties control metabolism are discussed in the light of the control analysis developed by H. Kacser and J. A. Burns ( (1973) in: Rate Control of Biological Processes, Davies DD, ed., Cambridge University Press, pp. 65-104) and R. Heinrich and T. A. Rapoport ( (1974) Eur. J. Biochem. 42, 89-95), and recent experimental data. Points at which the existing notions should be adjusted are: Metabolic control is shared by enzymes rather than confined to one rate-limiting enzyme per pathway. Whether an enzyme exercises strong control on a flux cannot be deduced solely from its own properties, nor is it directly related to its distance from equilibrium. With respect to metabolic control, enzymes should be classified into four groups, rather than two (reversible versus irreversible). The distribution of control among the enzymes depends on the metabolic conditions. Control structures of metabolic pathways probably differ with the function of that pathway.


A non-socially-sensitive predictive model of prostate cancer for Asian males with benign prostatic hyperplasia: A multi-site cross-sectional case-control study.
Benign prostatic hyperplasia (BPH) is common in aging Asian males and is associated with an excess risk of developing prostate cancer (PCa). However, discussions about socially-sensitive experiences such as sexual activity, which can significantly predict PCa risk, may be considered stigmatized in Asian culture. This study aimed to develop a predictive model for PCa risk in Asian males with BPH using non-socially-sensitive information. A cross-sectional case-control study, with PCa patients as the cases and remaining as the controls, was conducted on a cohort of Taiwanese males with BPH from four medical institutions. Patients who met the inclusion criteria were enrolled, excluding those aged over 86 years or who had received human papillomavirus (HPV) vaccination. Non-socially-sensitive variables such as obesity, occupational exposure, HPV infection, and PCa family history score (FH score) were included in a fully adjusted logistic regression model, and depicted using a nomogram. Among 236 BPH patients, 45.3% had PCa. Obesity, occupational exposure, HPV infection, and family history of PCa were significantly associated with PCa risk. The FH score (OR = 1.89, 95% CI = 1.03-3.47, P = 0.041) had the highest impact, followed by HPV infection (OR = 1.47, 95% CI = 1.03-2.11, P = 0.034), occupational exposure (OR = 1.32, 95% CI = 1.15-1.51, P <0.001), and obesity (OR = 1.22, 95% CI = 1.07-1.41, P = 0.005). The nomogram accurately depicted the predictive risk, and the model demonstrated robust performance compared to individual factors. In addition, the subgroup analysis results showed elderly age group could obtain more favorable predictive performance in our proposed model (AUC = 0.712). This non-socially-sensitive predictive model for PCa risk in Taiwanese males with BPH integrates multiple factors that could provide acceptable PCa risk-predictive performance, especially for elderly BPH patients over 70 years, aiding clinical decision-making and early cancer detection.


Characterization of seed storage proteins in Populus and their homology with Populus vegetative storage proteins.
We investigated the synthesis and accumulation of vegetative storage proteins (VSPs) in poplar plantlets and the homology between poplar seed storage proteins (SSPs) and VSPs. One-dimensional SDS polyacrylamide gel electrophoresis confirmed that both seed and vegetative storage proteins contained two predominant polypeptides of MW 32 and 36 kDa, but the subunit composition of the polypeptides differed. The 32- and 36-kDa polypeptides were highly abundant in basal leaves, stems, and roots of poplar plantlets. The 36-kDa subunit was synthesized in all plantlet tissues examined, but the 32-kDa subunit was not, suggesting that the 36-kDa polypeptide is a precursor of the 32-kDa polypeptide. The 36- and 32-kDa polypeptides of both SSPs and VSPs were glycosylated and both were found to be albumins. In addition, both polypeptides cross-reacted with a VSP antibody. Protein fingerprint patterns generated with two different proteolytic enzymes were identical for the 36-kDa polypeptide isolated from seeds or from stem tissue. Our study provides evidence that poplar SSPs and VSPs exhibit homology, and that expression is neither tissue-specific nor regulated solely by photoperiod.


Quantitative analysis of glutathione and cysteine S-conjugates of microcystin-LR in the liver, kidney and muscle of common carp (Cyprinus carpio) in Lake Taihu.
Tissue distribution of microcystin (MC)-LR-GSH, MC-LR-Cys and MC-LR of omnivorous fish in Lake Taihu was investigated. MC-LR and MC-LR-Cys were detected in liver, kidney and muscle. The concentration of MC-LR in liver and kidney was 0.052 μg g-1 DW and 0.067 μg g-1 DW, respectively. MC-LR-Cys appeared to be an important metabolite with average contents of 1.104 μg g-1 DW and 0.724 μg g-1 DW in liver and kidney, and the MC-LR-Cys/MC-LR ratio in liver and kidney reaching as high as 21.4 and 10.8. High MC-LR-Cys/MC-LR ratio and a significant correlation between MC-LR-Cys and MC-LR concentration in liver, suggest that liver is more active in detoxification of MC-LR by formation of MC-LR-Cys for omnivorous fish. Furthermore, there might be a balance between the accumulation and depuration/metabolism of MC-LR-Cys in kidney. The MC-LR-Cys can be formed in kidney directly, or transported from liver or other tissues, while the MC-LR-Cys in kidney might be dissociated to MC-LR or excreted. Although MC-LR and its metabolites were scarcely detected in muscle, it is necessary to investigate the distribution of toxic metabolites in edible muscle.


The roots of plant defenses: integrative multivariate analyses uncover dynamic behaviors of gene and metabolic networks of roots elicited by leaf herbivory.
High-throughput analyses have frequently been used to characterize herbivory-induced reconfigurations in plant primary and secondary metabolism in above- and below-ground tissues, but the conclusions drawn from these analyses are often limited by the univariate methods used to analyze the data. Here we use our previously described multivariate time-series data analysis to evaluate leaf herbivory-elicited transcriptional and metabolic dynamics in the roots of Nicotiana attenuata. We observed large, but transient, systemic responses in the roots that contrasted with the pattern of co-linearity observed in the up- and downregulation of genes and metabolites across the entire time series in treated and systemic leaves. Using this newly developed approach for the analysis of whole-plant molecular responses in a time-course multivariate data set, we simultaneously analyzed stress responses in leaves and roots in response to the elicitation of a leaf. We found that transient systemic responses in roots resolved into two principal trends characterized by: (i) an inversion of root-specific semi-diurnal (12 h) transcript oscillations and (ii) transcriptional changes with major amplitude effects that translated into a distinct suite of root-specific secondary metabolites (e.g. alkaloids synthesized in the roots of N. attenuata). These findings underscore the importance of understanding tissue-specific stress responses in the correct day-night phase context and provide a holistic framework for the important role played by roots in above-ground stress responses.


[1-year results after posterior lamellar keratoplasty with manually dissected donor tissue].
Posterior lamellar keratoplasty (PLK) is a reasonable alternative to penetrating keratoplasty (PK) for pathological changes of the corneal endothelium. The lamellar preparation of the donor tissue can be done in an automated fashion with a microkeratome, Descemet Stripping Automated Keratoplasty (DSAEK) or manually, Descemet Stripping Endothelial Keratoplasty (DSEK). Here, we describe our clinical results with one year follow-up using manually dissected donor lamellae for PLK. For 28 eyes (28 patients) best-corrected visual acuity, corneal topography, esthesiometry, pachymetry, as well as endothelial cell count were measured before, 1 day, 1 and 6 weeks, as well as 3, 6 and 12 months after surgery. 3 of 28 grafts showed primary graft failure. For 19 eyes (no other vision-limiting diseases), the best-corrected visual acuity rose on average from 1.0 +/- 0.53 to 0.4 +/- 0.20 (logMAR) 1 year after surgery. The surgically induced astigmatism changed from 3.8 +/- 2.8 dpt preoperatively to 1.4 +/- 1.0 dpt 12 months after DSEK. The pachymetry measured by slit lamp showed an averaged thickness of 649 +/- 54 mm pre- and 776 +/- 75 mm one week and 636 +/- 79 mm 1 year after surgery was performed. The endothelial cell count of the grafts was 2615 +/- 156 cells/mm (2), after one year postoperatively the mean endothelial cell count was 2084 +/- 536 cells/mm (2). The esthesiometry showed a slight rise from 0.96 +/- 0.34 g/mm (2) pre- to 0.96 +/- 0.03 g/mm (2) one year post-surgery. The postoperative course of our patients shows that improvement of visual acuity by PLK with manually dissected donor tissue is possible.


An analytical study of drug utilization, disease progression, and adverse events among 165 COVID-19 patients.
The coronavirus disease 2019 (COVID-19) epidemic has lasted for nearly 4 months by this study was conducted. We aimed to describe drug utilization, disease progression, and adverse drug events of COVID-19. A retrospective, single-center case series study enrolled 165 consecutive hospitalized COVID-19 patients who were followed up until March 25, 2020, from a designated hospital in Wuhan. Patients were grouped by a baseline degree of severity: non-severe and severe. An analytical study of drug utilization, disease progression, and adverse events (AEs) of COVID-19 was conducted. Of the 165 COVID-19 cases, antivirals, antibacterials, glucocorticoids, and traditional Chinese medicine (TCM) were administered to 92.7%, 98.8%, 68.5%, and 55.2% of patients, respectively. The total kinds of drugs administered to the severe subgroup [26, interquartile range (IQR) 18-39] were 11 more than the non-severe subgroup (15, IQR 10-24), regardless of comorbidities. The 2 most common combinations of medications in the 165 cases were 'antiviral therapy + glucocorticoids + TCM' (81, 49.1%) and 'antiviral therapy + glucocorticoids' (23, 13.9%). Compared with non-severe cases, severe cases received more glucocorticoids (88.5% vs. 66.2%, P=0.02), but less TCM (50.0% vs. 63.3%, P=0.20), and suffered a higher percentage of death (34.6% vs. 7.2%, P=0.001). At the end of the follow-up, 130 (78.8%) patients had been discharged, and 24 (14.5%) died. There were 13 patients (7.9%) who had elevated liver enzymes, and 49 patients (29.7%) presented with worsening kidney function during the follow-up. Of the 165 COVID-19 patients, the fatality rate remained high (14.5%). Drug utilization for COVID-19 was diverse and generally complied with the existing guidelines. Combination regimens containing antiviral drugs might be beneficial to assist COVID-19 recovery. Additionally, liver and kidney AEs should not be ignored.


Malignant papillary renal tumors with extensive clear cell change: a molecular analysis by microsatellite analysis and fluorescence in situ hybridization.
Histologic subtyping of renal cell carcinomas (RCCs) is based not only on cytoarchitectural pattern but also on distinct cytogenetic abnormalities. Some renal tumors demonstrate overlapping morphologic features, rendering histologic subtyping difficult. One such group of tumors is papillary renal neoplasms with extensive clear cell change. Because histologic subtyping has been shown to be of prognostic value, it is important that malignant epithelial renal tumors be accurately subtyped. It is not known if these tumors should be classified as papillary RCC (PRCC) or as conventional/(clear cell) RCC (CRCC). To ascertain if this subgroup of renal neoplasms demonstrates the cytogenetic abnormalities seen typically in PRCC, that is, trisomy 7 and 17 or CRCC, that is, loss of 3p, using microsatellite analysis for loss of heterozygosity (LOH), and fluorescence in situ hybridization (FISH) for trisomies. Seven RCCs from 6 patients that showed more than 75% papillary architecture and more than 75% clear cell change were included in the study. Tumor size ranged from 2.5 to 7.0 cm (mean 4.7 cm) and all were confined to the kidney (stage I). DNA was extracted from formalin-fixed paraffin-embedded tissue. FISH was done using In Situ Kits for centromere probes for chromosomes 7 and 17. For LOH, microsatellite analysis using labeled primers for 4 markers in the 3p13 through 3p24.2 region were used. The amplified polymerase chain reaction products were analyzed using an automated DNA sequencer. As compared with normal DNA, LOH in tumor was recognized as a loss of 1 allele, and microsatellite instability as the addition of an extra allele. LOH in at least 1 of the markers spanning for 3pl3 through 3p24.2 was detected in 6 of 7 specimens (86%), of which 1 also showed concomitant microsatellite instability. FISH did not demonstrate trisomy for either chromosome 7 or 17. Instead, monosomy 7 was observed in 4 of 6 tumors (67%) and monosomy 17 in all tumors (100%). Because malignant papillary renal tumors with extensive clear cell change show molecular changes identical to CRCC, this subgroup of tumors may have to be classified as CRCC. This study underscores the utility of molecular studies in refining light-microscopic criteria in accurate histologic subtyping of RCCs.


Effects of Intermittent Calf Muscle Electrical Stimulation on Walking Capacity in Claudicants Living with Type 2 Diabetes.
First-line therapy for diabetic patients presenting with intermittent claudication includes supervised exercise programs to improve walking distance. However, exercise comes with a variety of barriers and may be contraindicated in certain conditions. The aim of this study was to evaluate whether calf muscle electrostimulation improves claudication distance. A prospective, one-group, pretest-posttest study design was used on 40 participants living with type 2 diabetes mellitus, peripheral artery disease (ankle brachial pressure index, <0.90), and calf muscle claudication. Calf muscle electrostimulation of varying frequencies (1-250 Hz) was prescribed on both limbs for 1-hour daily sessions for 12 consecutive weeks. The absolute claudication distance (ACD) was measured at baseline and after the intervention. The recruited cohort (30 men and ten women; mean age, 71 years; mean ankle brachial pressure index, 0.70) registered a mean ± SD baseline ACD of 333.71 ± 208 m. After a mean ± SD of 91.68 ± 6.23 days of electrical stimulation, a significant mean ± SD increase of 137 ± 136 m in the ACD (P = .001, Wilcoxon signed rank test) was registered. Improvement was found to be sex independent, but age was negatively correlated with proportion of improvement (r = -0.361; P = .011, Pearson correlation test). Electrical stimulation of varying frequencies on ischemic calf muscles significantly increased the maximal walking capacity in claudicants with type 2 diabetes. This therapeutic approach should be considered in patients with impaired exercise tolerance or as an adjunct treatment modality.


Prevention and Management of Hypertension and Diabetes Using Social Capital and Physical Activity Among Socioeconomically Disadvantaged Populations.
The purpose of this study is to examine the association between physical activity adherence and social capital among uninsured primary care patients with or without hypertension and/or diabetes. Uninsured adults using a free clinic (N = 374) participated in a self-administered survey from January to April in 2016. The percentage of participants who were adherent to physical activity was low regardless of having hypertension and/or diabetes, or not. Individuals who reported a high sense of community were associated with a higher percentage of physical activity adherence. Good weight management was associated with less likelihood of physical activity adherence.


Poliovirus proteinase 3C: large-scale expression, purification, and specific cleavage activity on natural and synthetic substrates in vitro.
Proteinase 3C of poliovirus type 2 (Sabin) was expressed at 4% total protein in Escherichia coli. The protein was soluble and could be purified by a simple scheme. It was weakly active on the capsid precursor P1 (expressed in vitro), which contains two cleavage sites. The products of processing P1 were 1ABC and 1D (VP1). The activity was insensitive to Triton X-100. Crude extracts of cells infected with poliovirus type 1 (Mahoney) gave strong processing and yielded 1AB (VP0), 1C (VP3), and 1D in the same assay system but were sensitive to detergent. 3C from cell extracts that was separated from its precursors resembled the recombinant proteinase in its activity. Recombinant 3C cleaved the peptide dansyl-Glu-Glu-Glu-Ala-Met-Glu-Gln-Gly-Ile-Thr-Asn-Lys-NH2 at the Gln-Gly bond. We conclude that 3C is merely the core of the Gln-Gly-cleaving activity which processes P1 in vivo and that there is probably a hydrophobic contact between a larger 3C precursor and its P1 substrate which allows the second processing reaction: 1ABC, 1D----1AB, 1C, 1D.


Immunosuppressant-Related Lower Eyelid Edema in Transplant Patients.
Solid organ transplantation is the preferred method of treatment for a number of advanced medical conditions, but it requires systemic immunosuppression to prevent transplant rejection. The authors report 2 unique cases of persistent eyelid edema following solid organ transplantation believed to be related to their systemic immunosuppression. The eyelid findings developed after initiation of the immunosuppressant sirolimus. In 1 patient, the eyelid edema has persisted despite discontinuation of the medication. In the second patient, the immunosuppression could not be altered; therefore, he underwent surgical excision of the edematous lower eyelid. Sirolimus associated eyelid edema is an important medication side effect for ophthalmic and eyelid specialists to consider when a patient with a history of organ transplantation presents with localized noninflamed eyelid edema. This edema can persist despite discontinuation of the medication. Surgical excision of the edematous eyelid can achieve good results.


Configuration and interaction of the polar head group in gangliosides.
1. The interactions of gangliosides with Ca(2+) and some polar-head-group requirements for establishment of particular interactions with phosphatidylcholine were studied in monolayers at the air/145mm-NaCl interface. 2. Ganglioside-Ca(2+) interactions, as revealed by surface-potential measurements, depended on the position occupied by sialosyl residues in the oligosaccharide chain. The interactions with Ca(2+) of the single sialosyl residue of monosialogangliosides occurred above 0.1mm-CaCl(2), whereas the interaction of the cation with additional sialosyl groups in di- or tri-sialogangliosides depended on the carbohydrate residue to which the sialosyl moiety was attached. The sialosyl residue bound in sialosyl-sialosyl linkage interacted very little with Ca(2+). The sialosyl residue attached to the terminal galactose of the neutral tetrasaccharide chain interacted with Ca(2+) above 1mum-CaCl(2). 3. Experiments with mixed monolayers containing dihexadecyl phosphate and hexadecyltrimethylammonium indicated that for the occurrence of interactions of polysialogangliosides with phosphatidylcholine characterized by reductions in molecular packing and surface potential both charged groups of the phospholipid and sialosyl residues with particular dipolar properties in the ganglioside are participating. 4. Possible configurations that can explain the behaviour in monolayers were inspected with space-filling molecular models. The position of the carboxylate group of sialosyl residues with respect to the interface and to the sialosyl molecular plane can explain the different orientation of the dipole-moment vector of this residue, which depends on the position to which it is linked in the oligosaccharide chain. Favoured interactions of polysialogangliosides with phosphatidylcholine may result from a configuration allowing a partial matching of two oppositely oriented electrical vectors contributed by the zwitterionic phosphocholine group and particular sialosyl groups.


Psychosocial safety climate, emotional exhaustion, and work injuries in healthcare workplaces.
Preventing work injuries requires a clear understanding of how they occur, how they are recorded, and the accuracy of injury surveillance. Our innovation was to examine how psychosocial safety climate (PSC) influences the development of reported and unreported physical and psychological workplace injuries beyond (physical) safety climate, via the erosion of psychological health (emotional exhaustion). Self-report data (T2, 2013) from 214 hospital employees (18 teams) were linked at the team level to the hospital workplace injury register (T1, 2012; T2, 2013; and T3, 2014). Concordance between survey-reported and registered injury rates was low (36%), indicating that many injuries go unreported. Safety climate was the strongest predictor of T2 registered injury rates (controlling for T1); PSC and emotional exhaustion also played a role. Emotional exhaustion was the strongest predictor of survey-reported total injuries and underreporting. Multilevel analysis showed that low PSC, emanating from senior managers and transmitted through teams, was the origin of psychological health erosion (i.e., low emotional exhaustion), which culminated in greater self-reported work injuries and injury underreporting (both physical and psychological). These results underscore the need to consider, in theory and practice, a dual physical-psychosocial safety explanation of injury events and a psychosocial explanation of injury underreporting.


Asymmetric Michael Addition of 2-Acetyl Azaarenes to β-CF3-β-(3-indolyl)nitroalkenes Catalyzed by a Cobalt(II)/(imidazoline-oxazoline) Complex.
The first enantioselective Michael addition of 2-acetyl azaarenes to β-CF3-β-(3-indolyl)nitroalkenes has been successfully achieved in the presence of a Co(II)/(imidazoline-oxazoline) complex as the catalyst. The reaction affords a series of CF3- and 3-indole-containing adducts featuring a trifluoromethylated all-carbon quaternary stereocenter in good to excellent yields (up to 99%) and enantioselectivities (up to 96% ee). Furthermore, the functional groups in the adducts including C═O, NO2, and the azaarene provide a large variety of useful transformations, leading to the formation of valuable intermediates such as optically active secondary alcohol, pyrroline, ester, and pyrrolidinone which all contain a 3-substituted indole and a trifluoromethylated all-carbon quaternary stereocenter.


Laparoendoscopic single-site cholecystectomy in a pregnant patient.
Feasibility and safety of laparoscopic cholecystectomy during pregnancy for patients with symptomatic or complicated gallstone disease is well established. Laparoendoscopic single-site cholecystectomy (LESS-chole) is a new modality in which the entire surgery is undertaken via a transumbilical incision. We describe a 33-year-old patient who underwent a LESS-chole in the 20(th) week of pregnancy for gallstone disease complicated by episodes of obstructive jaundice and acute pancreatitis. This is the first reported case of LESS-chole performed using conventional laparoscopic instruments. The technical aspects as well as the various perioperative measures utilized to undertake this procedure safely are outlined.


Model-based Bayesian clustering (MBBC).
The program MBBC 2.0 clusters time-course microarray data using a Bayesian product partition model. The Bayesian product partition model in Booth et al. (2007) simultaneously searches for the optimal number of clusters, and assigns cluster memberships based on temporal changes of gene expressions. MBBC 2.0 to makes this method easily available for statisticians and scientists, and is built with three free computer language software packages: Ox, R and C++, taking advantage of the strengths of each language. Within MBBC, the search algorithm is implemented with Ox and resulting graphs are drawn with R. A user-friendly graphical interface is built with C++ to run the Ox and R programs internally. Thus, MBBC users are not required to know how to use Ox, R or C++, but they must be pre-installed. A self-extractable zip file, MBBC20zip.exe, is available at the MBBC webpage www.stat.ufl.edu/~casella/mbbc/, which contains MBBC.exe, source files, and all other related files. The current version works only in the Windows operating system. A free installation program and overview for Ox is available at www.doornik.com. A detailed installation guide for Ox is provided by MBBC, and is accessible without installing Ox. R is available at www.r-project.org/.


Photophysical properties of open-framework germanates templated by nickel complexes.
Open-framework germanates are a group of germanium oxides with a well-defined porous structure, suitable for ion-exchange and gas adsorption applications. Recently, Ni incorporation into the porous structure by establishing Ge-O-Ni bonds with the molecular complexes [Ni(H2N(CH2)2NH2)2] was realized. We investigate the optical and electronic features of these systems (SUT-1 and SUT-2) from first principles. To describe the photophysical behavior, we analyze the bonding between the Ni and nearest-neighboring atoms and simulate the absorption spectra. Because of their optical characteristics, germania-based nanomaterials are expected to be essential components of future optical and electronic devices. We discuss to what extent molecular transition-metal complexes embedded into porous germanium oxide can modify the optical response to potentially expand the area of applications.


Modulation of the respiratory supercomplexes in yeast: enhanced formation of cytochrome oxidase increases the stability and abundance of respiratory supercomplexes.
Yeast cells deficient in the Rieske iron-sulfur subunit (Rip1) of ubiquinol-cytochrome c reductase (bc1) accumulate a late core assembly intermediate, which weakly associates with cytochrome oxidase (CcO) in a respiratory supercomplex. Expression of the N-terminal half of Rip1, which lacks the C-terminal FeS-containing globular domain (designated N-Rip1), results in a marked stabilization of trimeric and tetrameric bc1-CcO supercomplexes. Another bc1 mutant (qcr9Δ) stalled at the same assembly intermediate is likewise converted to stable supercomplex species by the expression of N-Rip1, but not by expression of intact Rip1. The N-Rip1-induced stabilization of bc1-CcO supercomplexes is independent of the Bcs1 translocase, which mediates Rip1 translocation during bc1 biogenesis. N-Rip1 induces the stabilization of bc1-CcO supercomplexes through an enhanced formation of CcO. The association of N-Rip1 with the late core bc1 assembly intermediate appears to confer stabilization of a CcO assembly intermediate. This induced stabilization of CcO is dependent on the Rcf1 supercomplex stabilization factor and only partially dependent on the presence of cardiolipin. N-Rip1 exerts a related induction of CcO stabilization in WT yeast, resulting in enhanced respiration. Additionally, the impact of CcO stabilization on supercomplexes was observed by means other than expression of N-Rip1 (via overexpression of CcO subunits Cox4 and Cox5a), demonstrating that this is a general phenomenon. This study presents the first evidence showing that supercomplexes can be stabilized by the stimulated formation of CcO.


Effects of intensive exercise combined with dapagliflozin on body composition in patients with type 2 diabetes: a randomized controlled trial.
This study was aimed to evaluate the effects of intensive exercise in addition to the administration of sodium-glucose cotransporter 2 inhibitor dapagliflozin (DAPA) on body composition, including fat-free mass, in type 2 diabetes. We randomly assigned 146 patients to 24 weeks of treatment with intensive exercise, including resistance training, plus 5 mg (up to 10 mg) of DAPA daily (IT group) or DAPA alone (CT group). The primary endpoint was the difference in the change in fat-free mass from baseline to 24 weeks between the groups. The skeletal muscle mass index (SMI); metabolic profile, including HbA1c; and regional fat mass were also determined. ANCOVA was used for the group comparison, with least squares mean (LSM) differences and 95% confidence interval (CI). There was no significant difference in the change in fat-free mass (LSM difference -0.1 kg (95% CI: -0.5 to 0.4) and SMI (LSM difference -0.1 kg (95% CI: -0.2 to 0.1) between the groups. In contrast, the reduction of trunk fat mass was significantly higher in the IT group than in the CT group ((LSM difference -0.5 kg [95% CI -0.9 to -0.1]). Higher adherence to the resistance training tended to be associated with changes in HbA1c and high-sensitivity CRP levels. Our study suggests that intensive exercise do not prevent the reduction of fat-free mass after administration of SGLT2 inhibitors but can increase the reduction in abdominal fat, presumably leading to further improvements of hyperglycemia and chronic inflammation than DAPA alone in type 2 diabetes patients.


Histological evaluation of brain damage caused by crude quinolizidine alkaloid extracts from lupines.
The effects of the intracerebroventricular (ICV) administration of crude extracts of lupin quinolizidine alkaloids (LQAs) were studied in adult rat brain tissue. Mature L. exaltatus and L. montanus seeds were collected in western Mexico, and the LQAs from these seeds were extracted and analyzed by capillary gas chromatography. This LQA extract was administered to the right lateral ventricle of adult rats through a stainless steel cannula on five consecutive days. While control animals received 10 microl of sesame oil daily (vehicle), the experimental rats (10 per group) received 20 ng of LQA from either L. exaltatus or from L. montanus. All the animals were sacrificed 40 h after receiving the last dose of alkaloids, and their brains were removed, fixed and coronal paraffin sections were stained with haematoxylin and eosin. Immediately after the administration of LQA the animals began grooming and suffered tachycardia, tachypnea, piloerection, tail erection, muscular contractions, loss of equilibrium, excitation, and unsteady walk. In the brains of the animals treated with LQA damaged neurons were identified. The most frequent abnormalities observed in this brain tissue were "red neurons" with shrunken eosinophilic cytoplasm, strongly stained pyknotic nuclei, neuronal swelling, spongiform neuropil, "ghost cells" (hypochromasia), and abundant neuronophagic figures in numerous brain areas. While some alterations in neurons were observed in control tissues, unlike those found in the animals treated with LQA these were not significant. Thus, the histopathological changes observed can be principally attributed to the administration of sparteine and lupanine present in the alkaloid extracts.


Epidermal hepcidin is required for neutrophil response to bacterial infection.
Novel approaches for adjunctive therapy are urgently needed for complicated infections and patients with compromised immunity. Necrotizing fasciitis (NF) is a destructive skin and soft tissue infection. Despite treatment with systemic antibiotics and radical debridement of necrotic tissue, lethality remains high. The key iron regulatory hormone hepcidin was originally identified as a cationic antimicrobial peptide (AMP), but its putative expression and role in the skin, a major site of AMP production, have never been investigated. We report here that hepcidin production is induced in the skin of patients with group A Streptococcus (GAS) NF. In a GAS-induced NF model, mice lacking hepcidin in keratinocytes failed to restrict systemic spread of infection from an initial tissue focus. Unexpectedly, this effect was due to its ability to promote production of the CXCL1 chemokine by keratinocytes, resulting in neutrophil recruitment. Unlike CXCL1, hepcidin is resistant to degradation by major GAS proteases and could therefore serve as a reservoir to maintain steady-state levels of CXCL1 in infected tissue. Finally, injection of synthetic hepcidin at the site of infection can limit or completely prevent systemic spread of GAS infection, suggesting that hepcidin agonists could have a therapeutic role in NF.


The lymphocyte transformation test in allergic contact dermatitis: New opportunities.
Allergic contact dermatitis (ACD) is driven by the activation and proliferation of allergen-specific memory T-lymphocytes and is currently diagnosed by patch testing with a selected panel of chemical allergens. The lymphocyte transformation test (LTT) can be used to monitor ex vivo T-lymphocyte responses to antigens, including contact allergens. The LTT is not viewed as being an alternative to patch testing, but it does seek to reflect experimentally skin sensitization to specific chemicals. The LTT is based on stimulation in vitro of antigen-driven T-lymphocyte proliferation. That is, exposure in culture of primed memory T-lymphocytes to the relevant antigen delivered in an appropriate configuration will provoke a secondary response that reflects the acquisition of skin sensitization. The technical aspects of this test and the utility of the approach for investigation of immune responses to contact allergens in humans are reviewed here, with particular emphasis on further development and refinement of the protocol. An important potential application is that it may provide a basis for characterizing those aspects of T-lymphocyte responses to contact allergens that have the greatest influence on skin sensitizing potency and this will be considered in some detail.