Chromogranin A and the tumor microenvironment.
Chromogranin A (CgA) is an acidic glycoprotein belonging to a family of regulated secretory proteins stored in the dense core granules of the adrenal medulla and of many other neuroendocrine cells and neurons. This protein is frequently used as a diagnostic and prognostic serum marker for a range of neuroendocrine tumors. Circulating CgA is also increased in patients with other diseases, including subpopulations of patients with non-neuroendocrine tumors, with important prognostic implications. A growing body of evidence suggests that CgA is more than a diagnostic/prognostic marker for cancer patients. Indeed, results of in vitro experiments and in vivo studies in animal models suggest that this protein and its fragments can affect several elements of the tumor microenvironment, including fibroblasts and endothelial cells. In this article, recent findings implicating CgA as a modulator of the tumor microenvironment and suggesting that abnormal secretion of CgA could play important roles in tumor progression and response to therapy in cancer patients are reviewed and discussed.


Mechanisms of deposition of epicuticular wax in leaves of broccoli, Brassica oleracea var. capitata L.
Broceeli (Brassika oleraceo L. var. capitata L.) leaf waxes were labelled with [2-14 Cjacetate and examined during synthesis and deposition using microautoradiography. Waxes were viewed with light microscopy and scanning electron microscopy (SEM), including cryo-SEM. In microautoradiography, silver grains resulting from radioactive decay of labelled waxes seemed to be distributed randomly throughout the cuticle. This random distribution supports the hypothesis of diffusion or complex anastomosing pores as the mechanism of epicuriculur wax position. The hypothesis of wax-depositing pores running perpendicular to the leaf surface is not supported.


The outward transport of catecholamines mediated by uptake2 of the rat heart.
The efflux of 3H-catecholamines from the extra-neuronal tissue of the rat heart was analysed (after inhibition of vesicular and neuronal uptake, monoamine oxidase and catechol-O-methyl transferase). In most experiments, hearts were first loaded with a tracer concentration of a 3H-catecholamine and then washed out. For all four catecholamines [3H-(+/-)-isoprenaline, 3H-(+/-)-adrenaline, 3H-(-)-noradrenaline, and 3H-dopamine] the loading period resulted in virtually the same distribution pattern: most of the radioactivity distributed into "compartment III". However, the rate constants for efflux from compartment III increased in the order 3H-(-)-noradrenaline less than 3H-dopamine less than 3H-(+/-)isoprenaline = 3H-(+/-)-adrenaline. O-methyl-isoprenaline (OMI, a potent inhibitor of uptake2) caused a concentration-dependent and partial inhibition of the efflux of all 3H-catecholamines; its IC50 (half-maximal inhibition of OMI-sensitive efflux) was very close to that for half-maximal inhibition of inward transport by uptake2. It is concluded that there is not only (OMI-resistant) diffusional efflux of 3H-catecholamines, but also (OMI-sensitive) outward transport of 3H-catecholamines. The contribution by each of these processes to total efflux differed considerably from one 3H-catecholamine to the next. U-0521 (the COMT inhibitor used in this study) inhibited the OMI-sensitive efflux of 3H-noradrenaline with an IC50 of about 100 mumol/l. However, no inhibitory effect was found for 10 mumol/l U-0521. During the wash-out period (see above) various unlabelled substrates of uptake2 were added to the perfusion fluid at a concentration equalling 2 X Km.(ABSTRACT TRUNCATED AT 250 WORDS)


Minimally invasive synovium harvest for potential use in meniscal tissue engineering.
Tissue engineering is being investigated as a means for treating avascular meniscal injury or total meniscal loss in human and veterinary patients. The purpose of this study was to determine if an arthroscopic tissue shaver can be used to collect viable synoviocytes for in vitro culture during therapeutic stifle arthroscopy, with the long term goal of producing autologous meniscal fibrocartilage for meniscal tissue engineering. Synovium was harvested arthroscopically from 13 dogs with naturally occurring cranial cruciate ligament deficiency and obtained from 5 dogs with patellar luxation via arthrotomy. Cells harvested via arthroscopy and arthrotomy were treated with a chondrogenic growth factor protocol and analyzed for meniscal-like matrix constituents including collagens type I, II, and glycosaminoglycans. Arthrotomy and Arthroscopic origin cells formed contracted tissues containing collagen I, II and small amounts of GAG. These surgical methods provide clinically relevant access to synoviocytes for potential use in meniscal tissue engineering.


Eicosapentaenoic acid stimulates leptin receptor gene expression in the hypothalamus of newborn rats.
Body weight and obesity are controlled by the binding leptin (Ob) receptor, but in newborn rats, despite high Ob levels, hypothalamic leptin receptors (Ob-Rb) are only weakly expressed. In this study we have attempted to stimulate expression of the Ob-Rb gene by administering 2 polyunsaturated fatty acids (PUFAs) recommended for the maternal diet and known as gene regulators: docosahexaenoic acid and eicosapentaenoic acid (EPA). We studied the effects of a single dose injected into a cerebral ventricle of newborn rats on postnatal day 2. On days 1, 2, and 3 after administration, we dissected the hypothalamus and analyzed Ob-Rb and Ob messenger RNAs by polymerase chain reaction and protein expression by Western blot immunoassay. Our results demonstrate that EPA, but not docosahexaenoic acid, caused an early messenger RNA expression of the gene, 24 hours earlier than in the controls, and the protein was also detected earlier. Our data corroborate the observations regarding the role of PUFAs, EPA in particular, in the regulation of gene expression. In addition, they support the recommendation to enrich the maternal diet with fish and seafood rich in n-3 PUFA, because the concentrations of n-6 and n-3 PUFA in human milk reflect the composition of fat in the mother's diet.


Unsupervised machine learning model to predict cognitive impairment in subcortical ischemic vascular disease.
It is challenging to predict which patients who meet criteria for subcortical ischemic vascular disease (SIVD) will ultimately progress to subcortical vascular cognitive impairment (SVCI). We collected clinical information, neuropsychological assessments, T1 imaging, diffusion tensor imaging, and resting-state functional magnetic resonance imaging from 83 patients with SVCI and 53 age-matched patients with SIVD without cognitive impairment. We built an unsupervised machine learning model to isolate patients with SVCI. The model was validated using multimodal data from an external cohort comprising 45 patients with SVCI and 32 patients with SIVD without cognitive impairment. The accuracy, sensitivity, and specificity of the unsupervised machine learning model were 86.03%, 79.52%, and 96.23% and 80.52%, 71.11%, and 93.75% for internal and external cohort, respectively. We developed an accurate and accessible clinical tool which requires only data from routine imaging to predict patients at risk of progressing from SIVD to SVCI. Our unsupervised machine learning model provides an accurate and accessible clinical tool to predict patients at risk of progressing from subcortical ischemic vascular disease (SIVD) to subcortical vascular cognitive impairment (SVCI) and requires only data from imaging routinely used during the diagnosis of suspected SVCI. The model yields good accuracy, sensitivity, and specificity and is portable to other cohorts and to clinical practice to distinguish patients with SIVD at risk for progressing to SVCI. The model combines assessment of diffusion tensor imaging and functional magnetic resonance imaging measures in patients with SVCI to analyze whether the "disconnection hypothesis" contributes to functional and structural changes and to the clinical presentation of SVCI.


Forgotten Joint Score for early outcome assessment after total knee arthroplasty: Is it really useful?
Forgotten Joint Score (FJS) has become a popular tool for total knee arthroplasty (TKA), but almost all studies had assessment performed 1 year after surgery. There is a need for a sensitive tool for earlier outcome assessment. The aim of this study was to investigate the usefulness of FJS within the first year after TKA. This was a cross-sectional study. Patients within the first year after primary TKA were recruited. FJS was translated into the local language with a cross-cultural adaptation and was validated by assessing the correlation with the Western Ontario and McMaster Universities Arthritis Index score (WOMAC). Ceiling and floor effects (highest or lowest 10% or 15%) of both scores were compared. Skewness of scores was assessed with a histogram. One hundred sixty-three subjects were recruited: 84 (51.5%) had evaluation at 3 months after the operation, 56 (34.4%) at 6 months, and 23 (14.1%) at 12 months. FJS had fewer patients at the highest 10% (10.7% vs. 16.1%, P = 0.046) or 15% (19.6% vs. 32.1%, P = 0.027) at 6 months and within the first year overall (6.7% vs. 13.5%, P <0.001; 14.1% vs. 22.7%, P <0.001). Also, it had more patients at the lowest 10% (16.7% vs. 0%, P <0.001) or 15% (21.4% vs. 0%, P <0.001) at 3 months, 6 months (10.7% vs. 0%, P <0.001), and overall (12.9% vs. 0%, P <0.001; 16.6% vs. 0%, P <0.001). The skewness was much less than WOMAC (0.09 vs. -0.56). FJS has a low ceiling effect but a high floor effect in the first year after TKA. Such characteristics make it less useful for the general assessment of early patient report outcome after operation.


Occupational contact dermatitis.
The two commonest forms of occupational skin disease are irritant and allergic contact dermatitis. Morphology and history are used to establish the diagnosis and the relation of the disorder to work exposure, respectively. Diagnostic patch tests are used to define further the diagnosis and aetiology of the dermatitis.


Dataset on nicotine-free, nontransgenic tobacco (Nicotiana tabacuml.) edited by CRISPR-Cas9.
This dataset in brief is related to the research letter entitled "Nicotine-free, nontransgenic tobacco (Nicotiana tabacuml.) edited by CRISPR-Cas9" [1]. Cured tobacco products with a significantly reduced nicotine content helps people to overcome their nicotine addiction. Here we summarize additional data and method descriptions of the generation process of a nicotine-free, nontransgenic tobacco plant. This included the cloning, transformation and regeneration of transgenic tobacco plants, followed by the analysis of the nicotine content and genomic modifications caused by CRISPR-Cas9 mediated gene editing. Subsequently, nicotine-free plants were screened for loss of T-DNA cassette, i.e. nontransgenity. Finally, a metabolic footprint was recorded by 1H NMR analysis.


Cell cycle control by homeobox genes in development and disease.
Homeobox-containing transcription factors are among the most studied players during normal embryonic development. In the last few years, they have also been shown to act as modulators of cell proliferation. In the present review, we highlight recent studies that demonstrated the influences of homeobox genes on crucial components of the cell cycle machinery. Homeobox genes may have direct or indirect influence on their transcription levels but can also interact with those components via protein-protein interaction. In many cases, these studies were done in the context of pathological states like cancer as well as during vertebrate development.


Treating the Chronic Disease of Obesity.
Obesity is a treatable chronic disease. Primary care providers play an essential role in diagnosis, treatment, and comprehensive care of patients with obesity. In recent years, treatment approaches have rapidly evolved, increasing effective and safe therapies. In this review, we provide practical information on the care of patients with obesity with a focus on antiobesity pharmacotherapy within the context of currently available therapeutic modalities such as intensive lifestyle interventions and bariatric surgery.


Exploration of doctor and patient agendas in general practice consultations.
A method for the interactional analysis of doctor/patient consultations is described and applied to six naturally occurring general practice interviews by three raters. The method is reliable given sufficient training and satisfies the stringent criteria for any method of analysis for medical interviews proposed by Wasserman and Inui [1: Wasserman R.C. and Inui R.S. Med. Care 21, 279, 1983].


Primary non-Hodgkins lymphoma of the ovary in the background of human immunodeficiency virus (HIV): a bold and curative approach to treatment.
Non-Hodgkins lymphoma of the ovary is a rare disease and there is only one previously documented case arising in a patient with human immunodeficiency virus (HIV). In this report, the authors discuss the management of a case of non-Hodgkins Lymphoma of the ovary occurring in a patient with HIV and demonstrate that treatment regimens may be successfully implemented in this immunocompromised population without an increase in adverse effects.


Aging well--the role of minimally invasive aesthetic dermatological procedures in women over 65.
The western world is getting older. Aging well has become the new target of preventative medicine. Aesthetic dermatology can contribute to this quest. Females over 65 represent an important and growing group of consumers of cosmetic procedures. In this group, there is a paucity of scientific evaluation of aesthetic procedures. To review the use of minimally invasive procedures for facial rejuvenation in women over 65. Drawing from both the literature and personal experience, the opportunities, modifications, and limitations of minimally invasive techniques for facial rejuvenation in older women are considered. In this older age group, dermal fillers, chemical and laser peels, and nonablative photorejuvenation remain useful and can each be used as stand-alone treatments. In this age group, botulinum toxin (BTX) injections are more often used in combination with other procedures. With respect to aesthetic procedures, women over 65 are different from younger women. More scientific investigation is necessary to better meet needs of this growing part of the population. Available data suggest that aesthetic dermatology can make a major contribution to the complex matter of aging well.


The Use of Concentrates Rich in Orange By-Products in Goat Feed and Its Effects on Physico-Chemical, Textural, Fatty Acids, Volatile Compounds and Sensory Characteristics of the Meat of Suckling Kids.
We analysed how replacing cereal concentrates with dehydrated orange pulp (DOP) in the diet of mother goats affects the meat quality of suckling kids. Three experimental diets for mother goats were designed. The DOP-0 diet contained commercial concentrates and alfalfa hay. In the DOP-40 and DOP-80 diets, 40% and 80% (respectively) of the cereal in the concentrate was replaced with pellets of DOP (the alfalfa hay component was unchanged). We evaluated the chemical composition, texture, water holding capacity, colour, fatty acids (FAs) profile, volatile compounds, and sensorial appraisal of the meat from 30 male suckling kids (cold carcass weight 4.74 kg, 4.82 kg, and 4.65 kg for DOP-0, DOP-40, and DOP-80, respectively) of the Payoya breed (n = 10 for each diet). Meat from kids in the DOP-40 and DOP-80 groups exhibited characteristics favourable for human health, including the meat's thrombogenicity index, PUFA/SFA ratio (0.60 index), and n-6 PUFA/n-3 PUFA ratio (approximately 7.50). The meat also exhibited reduced MUFA content (around 460 mg/100 g fresh meat). An increase in ethyl furan, dimethyl disulphide and heptane was observed in grilled meat from goats that were fed using DOP. The inclusion of DOP in goat feed improved consumers' sensory appreciation of the kid's meat.


[Quality control in the therapy of chronic pain. Results obtained by a task force of the German Section of the International Association for the Study of Pain on psychological assessment of chronic pain. XI. Assessment and multiaxial classification of pain].
This paper reviews instruments in German language for the psychological assessment and classification of pain. Usually chronic pain syndromes are classified within the International Classification of Diseases (ICD). Instead of the psychiatric chapter of the ICD, it is possible to use the Diagnostic and Statistical Manual of Mental Disorders (DSM). The proposed classification system of the International Association of the Study of Pain (IASP) is based on a multiaxial solution. The numerous ways to classify chronic pain include many problems and limits, especially in the case of an interdisciplinary assessment. They provide no specific system for classifying pain syndromes. They are impractical to handle and restrict classifying pain as either somatogenic or psychogenic. It is not possible to describe both sides in one diagnosis without loss of information. As a result of this situation, a task force of the German Chapter of the International Association for the Study of Pain developed a Multiaxial Classification of Pain (Multiaxiale Schmerzklassifikation; MASK) as an advanced system of pain assessment and an alternative to the common classification systems. MASK embraces a somatic (MASK-S) and a psychological (MASK-P) part. Both parts constitute an 'interdisciplinary diagnosis'. MASK-S enables classifying a pain syndrome using hierarchical levels, with progredient specific degrees of differentiation. In addition there are 6 axes to describe pain syndromes qualitatively and quantitatively (e.g. localization, quality). The psychosocial part (MASK-P) embraces 5 main levels (1. behavioral, 2. emotional, 3. cognitive, 4. stress-related, 5. habitual personal factors) and 2 additional levels (6. functional coherence, 7. ICD/DSM-diagnosis additional). The MASK-P part of the diagnosis is composed of graduation on these levels. Differential axis of the pain syndromes are described phenomenologically and specifically. MASK provides the possibility of establishing an integrative, interdisciplinary diagnosis.


Targeted codelivery of doxorubicin and IL-36γ expression plasmid for an optimal chemo-gene combination therapy against cancer lung metastasis.
Cancer metastasis is the main cause for the high mortality in breast cancer patients. In this work we developed a polymer POEG-st-Pmor for targeted co-delivery of IL-36γ expression plasmid and doxorubicin (Dox) to lung metastasis of breast cancer. The polymer readily formed micelles that were effective in loading Dox and simultaneously forming complexes with IL-36γ plasmid. Interestingly, particles co-loaded with Dox and plasmid was significantly smaller and more stable than the particles loaded with Dox only. Gene transfection in both lungs and s.c. tumors was significantly higher with our polymer compared to PEI. In addition, the Dox + IL-36γ/POEG-st-Pmor not only could bring improved anti-metastatic effect but synergistically enhance the type I immune response by increasing the IFN-γ positive CD4+ and CD8+ T cells and simultaneously decreasing the immunosuppressive myeloid-derived suppressor cells in the lung. POEG-st-Pmor may represent a simple and effective delivery system for an optimal chemo-gene combination therapy.


Discriminative stimulus effects of N-allylnormetazocine in rats trained to discriminate a kappa from a sigma agonist.
Rats were trained to discriminate ethylketocyclazocine (EKC) from vehicle, phencyclidine (PCP) from vehicle, or ethylketocyclazocine from phencyclidine on a two-lever operant task with a fixed-ratio 30 schedule of food-reinforcement on the appropriate lever. The three groups were tested with the training drugs (i.e., EKC and PCP), N-allylnormetazocine (NANM), and the (+)- and (-)-isomers of N-allylnormetazocine. EKC produced EKC-appropriate responding in the EKC-vehicle group and in the EKC-PCP group; it produced vehicle-appropriate responding in the PCP-vehicle group. Similarly, PCP produced PCP-appropriate responding in the PCP-vehicle group and in the EKC-PCP group but vehicle-appropriate responding in the EKC-vehicle group. The (+)-isomer of NANM produced PCP-appropriate responding in both the PCP-VEH and EKC-PCP groups, whereas the (-)-isomer produced EKC appropriate responding in the EKC-VEH and EKC-PCP groups. The results of this study demonstrate that rats can be trained to discriminate a kappa-agonist from a PCP/sigma-agonist and can differentiate these discriminative stimulus properties of other test compounds. These results also indicate that the (-)-isomer of NANM has kappa-agonist discriminative stimulus properties, whereas PCP/sigma-like effects are produced by the (+)-isomer.


Human Trophectoderm Spheroid Derived from Human Embryonic Stem Cells.
The use of human embryos for studying the early implantation processes and trophoblast is restricted by ethical concerns. The development of models mimicking the peri-implantation embryos is critical for understanding the physiology of human embryos and many pathophysiological disorders including recurrent implantation failure and miscarriage. Three-dimensional (3D) models of trophoblastic spheroids have been successfully derived from human embryonic stem cells (hESC). Simultaneous activation of the BMP pathway and blockage of the Activin/Nodal pathway favor the differentiation of hESC into trophoblast. Here we describe a 3D trophectoderm differentiation protocol with the use of BAP (BMP4, A83-01, and PD173074) to generate hESC-derived trophectoderm spheroids (BAP-EB). BAP-EB is highly reproducible and exhibits morphological and transcriptomic similarities to human early blastocysts.


Uprooting defects to enable high-performance III-V optoelectronic devices on silicon.
The monolithic integration of III-V compound semiconductor devices with silicon presents physical and technological challenges, linked to the creation of defects during the deposition process. Herein, a new defect elimination strategy in highly mismatched heteroepitaxy is demonstrated to achieve a ultra-low dislocation density, epi-ready Ge/Si virtual substrate on a wafer scale, using a highly scalable process. Dislocations are eliminated from the epilayer through dislocation-selective electrochemical deep etching followed by thermal annealing, which creates nanovoids that attract dislocations, facilitating their subsequent annihilation. The averaged dislocation density is reduced by over three orders of magnitude, from ~108 cm-2 to a lower-limit of ~104 cm-2 for 1.5 µm thick Ge layer. The optical properties indicate a strong enhancement of luminescence efficiency in GaAs grown on this virtual substrate. Collectively, this work demonstrates the promise for transfer of this technology to industrial-scale production of integrated photonic and optoelectronic devices on Si platforms in a cost-effective way.


Creating Community and Engaging Community: The Foundations of the Estate Little Princess Archaeology Project in St. Croix, United States Virgin Islands.
This article discusses how Co-Principal Investigators that designed and executed the Estate Little Princess Archaeology Project (ELPAP) came together as a community, to demonstrate how such a formation within the discipline, with all its ups and downs, facilitates the skills needed to conduct community archaeology. By using the ELPAP as a case study, this article provides a multiscale examination of the ELPAP, expanding the discourse on community archaeology to include community building practices among archaeologists, between organizations, and with communities impacted by archaeological work.


The Fluctuations of Blood Oxygen Level-Dependent Signals as a Method of Brain Tumor Characterization: A Preliminary Report.
In this study we present the nature and characteristic of the fluctuation of blood oxygen level-dependent (BOLD) signals measured from brain tumors. Supratentorial astrocytomas, which were neither operated nor previously managed with chemotherapy or radiotherapy, were segmented, and the time series of the BOLD signal fluctuations were extracted. The mean (across patients) power spectra were plotted for the different World Health Organization tumor grades. One-way analysis of variance (ANOVA) was performed to identify significant differences between the power spectra of different tumor grades. Results were considered significant at P < 0.05. A total of 58 patients were included in the study. This group of patients included 1 patient with grade I glioma; 15 with grade II; 12 with grade III; and 30 with grade IV. The power spectra of the tumor time series were individually inspected, and all tumors exhibited high peaks at the lower frequency signals, but these were more pronounced in high-grade tumors. ANOVA showed a significant difference in power spectra between groups (P = 0.000). Post hoc analysis with Bonferroni correction showed a significant difference between grade II and grade III (P = 0.012) and grade IV (P = 0.000). There was no significant power spectra difference between grade III and IV tumors (P = 1). The power spectra of BOLD signals from tumor tissue showed fluctuations in the low-frequency signals and were significantly correlated with tumor grade. These signals could have a misleading effect when analyzing resting state functional magnetic resonance imaging and could be also viewed as a potential method of tumor characterization.


Effect of counterion on the structural switchover and binding of piroxicam with sodium dodecyl sulfate (SDS) micelles.
Addition of an electrolyte such as NaCl to ionic micelles such as sodium dodecyl sulfate (SDS) alters the ionic atmosphere of the bulk solvent, thereby changing both the micellar properties and the interaction pattern of micelles with a molecule in the solvent. In this study, we show how added NaCl in the presence of SDS micelles modulates the surface charge of the micelles, which in turn fine-tunes the switchover equilibrium between anionic and global neutral forms of piroxicam. The presence of salt alters the CMC and aggregation number of SDS micelles. The binding of the global neutral form of piroxicam with SDS is found to be strongly modulated by the presence of counterion in the bulk solvent.


Efficacy of bacteriocin-containing cell-free culture supernatants from lactic acid bacteria to control Listeria monocytogenes in food.
Consumer demands have led to an increased interest in the use of natural antimicrobials for food protection. With the objective of developing novel products for enhancing the microbial safety of food, we have tested cell-free culture supernatants (CFS's) of eight antagonistic bacterial strains for their efficacy to inhibit Listeria monocytogenes in different food matrices. The antagonistic strains represented different members of the order Lactobacillales as well as one isolate of Staphylococcus sciuri and all showed strong inhibition of L. monocytogenes on agar plates. Cell-free supernatants were obtained after growing the bacteria in a yeast extract-glucose broth. In six of the CFS's, different class IIa bacteriocins, namely leucocin A, leucocin B, mundticin L, pediocin PA-1, sakacin A, and sakacin X, were identified as the major anti-listerial compounds. For the other two strains, the active substances could not be ascertained conclusively. The minimal effective concentration (MEC) of the individual CFS's to achieve a 2.3 log(10) reduction of L. monocytogenes was determined in culture broth, whole milk, and ground beef at 4°C. While all bacteriocin-containing CFS's were effective in broth at concentrations from 52 to 205 AU/ml, significant higher concentrations were needed when applied in food. Best results were obtained using CFS's containing pediocin PA-1, that displayed only three- and ten-times higher MEC's in milk (307 AU/ml) and ground meat (1024 AU/g) compared to broth, respectively. A twenty-fold increase in the MEC (2048 AU/ml) was observed for a mundticin L-containing fermentate, and a CFS containing leucocin A and B was inactivated more than fifty-fold (>1280 AU/ml) in both food matrices. Remarkably, the sakacin A and sakacin X containing CFS's displayed very selective inactivation rates, in which sakacin A was only effective in meat (512 AU/g), while sakacin X was only effective in milk (2048 AU/ml). In all cases, inhibition of L. monocytogenes was only transient and surviving or resistant bacteria started growing after prolonged storage. These results highlight the importance of careful testing the effectiveness of bacteriocins in the food systems for which they are intended to be applied against the selected target and non-target bacteria. Furthermore, the outgrowth of surviving or resistant bacterial populations points out that the tested bacteriocins are not suited to assure full inhibition of L. monocytogenes in a food product, if not applied in combination with additional preservative measures.


Bilateral destruction of neocortical and perirhinal projection targets of the acoustic thalamus does not disrupt auditory fear conditioning.
The present study examined whether complete bilateral destruction of auditory cortex would interfere with auditory fear conditioning in rats. Complete destruction of auditory cortex required lesions of temporal neocortical and perirhinal periallocortical areas. Fear conditioning was assessed by measuring freezing and arterial pressure responses elicited by an acoustic stimulus after pairing with footshock. Animals with complete bilateral lesions of auditory cortex showed conditioned arterial pressure and freezing responses comparable to those of unoperated controls. In contrast, bilateral destruction of the acoustic thalamus interfered with the conditioning of both responses. These results demonstrate that the auditory cortex is not required for the conditioning of fear responses to simple acoustic stimuli and add to the growing body of evidence that fear conditioning can be mediated by subcortical (amygdaloid) projections of the acoustic thalamus.


Prolonged haloperidol and reduced haloperidol plasma concentrations after decanoate withdrawal.
Haloperidol and reduced haloperidol plasma concentrations were measured in twelve schizophrenic patients upon cessation of haloperidol decanoate (HLD) treatment. Each patient received HLD 100 mg every 4 weeks for five injections. After the fifth injection, HLD was discontinued. Haloperidol and reduced haloperidol plasma concentrations were obtained prior to cessation and at weeks 1, 3, 4, 5, 7, 9, 11, and 13 post-injection. Haloperidol and reduced haloperidol plasma concentrations were assayed by HPLC. Both haloperidol and reduced haloperidol plasma concentrations were detectable 13 weeks post HLD discontinuation. Maximal haloperidol plasma concentrations were observed at one week post cessation and gradually declined. The mean elimination half-life for haloperidol was 27.4 +/- 8.6 days (range 19.0-47.0 days). Reduced haloperidol plasma concentrations declined very slowly. Our results show that both haloperidol and reduced haloperidol plasma concentrations can remain for extended time periods after HLD is discontinued.


Locus of control as a mediator of negative divorce-related events and adjustment problems in children.
Beliefs about whether locus of control mediates the relationship between negative divorce-related events and children's adjustment were explored in a group of 78 children and their primary residential parents. In the children's self-reports of adjustment, locus of control was found to be a partial mediator in the relationship. Analyses of the parents' reports of the children's adjustment did not support a mediational model.


Content of 5-alpha-reductase (type 1 and type 2) mRNA in dermal papillae from the lower abdominal region in women with hirsutism.
Androgens influence the growth of terminal hair. The dermal papilla contains androgen receptors and the enzymes 5-alpha-reductase types 1 and 2. Both of these enzymes convert testosterone to the more active androgen, 5-alpha-dihydrotestosterone. The male distribution pattern of terminal hair in females is termed hirsutism. It is most common among women with hyperandrogenism; however, it may also affect patients with normal androgen levels (idiopathic hirsutism). The aim of this study was to assess the expression of 5-alpha-reductase types 1 and 2 mRNA in dermal papillae from the lower abdominal skin in women with hirsutism. The study included 42 subjects, 24 with a diagnosis of polycystic ovary syndrome (PCOS) and 18 with idiopathic hirsutism (IH). In all patients, free serum testosterone was measured. The mean +/- SD concentration of free serum testosterone was 7.2 +/-5.3 pmol/L in the total group of patients, 10.8 +/- 4.0 pmol/L in patients with PCOS, and 2.5 +/- 1.7 pmol/L in patients with IH. Quantitative analysis was then performed for the mRNA of 5-alpha-reductase types 1 and 2, both of which were found within the dermal papillae from the lower abdominal skin region. The number of mRNA copies/microg of total RNA for 5-alpha-reductase type 1 was statistically significantly higher than that for type 2 in both groups of examined patients. We also demonstrated a positive correlation between the number of mRNA copies/microg of total RNA for 5-alpha-reductase types 1 and 2 and the concentration of free serum testosterone in women with PCOS and IH. Considering all patients together, we found a positive correlation between the number of mRNA copies/microg of total RNA for 5-alpha-reductase type 2 and the concentration of free serum testosterone. There was also a tendency towards a positive correlation between the number of mRNA copies/microg of total RNA for 5-alpha-reductase type 1 and the concentration of free serum testosterone. The results of our study suggest that testosterone increases expression of 5-alpha-reductase types 1 and 2 in dermal papillae from the lower abdominal region in patients with hirsutism.


Finger and forearm vasodilatatory changes after local cold acclimation.
To determine the vascular changes induced by local cold acclimation, post-ischaemia and exercise vasodilatation were studied in the finger and the forearm of five subjects cold-acclimated locally and five non-acclimated subjects. Peak blood flow was measured by venous occlusion plethysmography after 5 min of arterial occlusion (PBFisc), after 5 min of sustained handgrip at 10% maximal voluntary contraction (PBFexe), and after 5 min of both treatments simultaneously (PBFisc + exe). Each test was performed at room temperature (25 degrees C, SE 1 C) (non-cooled condition) and after 5 min of 5 degrees C cold water immersion (cooled condition). After the cold acclimation period, the decrease in skin temperature was more limited in the cold-acclimated compared to the non-acclimated (P less than 0.01). The PBFisc was significantly reduced in the cooled condition only in the cold-acclimated subjects (finger: 8.4 ml.100 ml-1.min-1, SE 1.1, P less than 0.01; forearm: 5.8 ml.100 ml-1.min-1, SE 1.5, P less than 0.01) compared to the non-cooled condition. Forearm PBFexe was significantly decreased in the cooled condition only in the cold-acclimated subjects (non-cooled: 7.4 ml.100 ml-1.min-1, SE 1.2; cooled: 3.9 ml.100 ml-1.min-1, SE 2.6, P less than 0.05) indicating that muscle blood flow was also reduced.(ABSTRACT TRUNCATED AT 250 WORDS)


Causes of laryngeal cartilage and hyoid bone fractures found at postmortem.
The principal aims of this retrospective study were to assess the prevalence and causes of laryngo-hyoid fractures found in coroners' adult postmortem examinations over a five-year period. In 78 of 1930 cases (4%), there was a fracture of the larynx (thyroid or cricoid cartilage) or hyoid bone. The thyroid cartilage alone was fractured in 38 cases; the hyoid bone alone was fractured in 19 cases; the larynx and hyoid bone were both fractured in 21 cases, including one which involved the cricoid cartilage. There was no evidence of surface injury to the neck in 14.1% of cases. The majority (44/78; 56.4%) involved classical manual or ligature 'pressure to the neck', i.e. hanging (32) or strangulation (12) but a significant minority (35.9%) involved other circumstances: road/rail traffic collision (12; 15.4%); falls (6; 7.7%); assaults involving blunt force trauma to the head and neck (4; 5.1%); incised wounds (3; 3.8%); gunshot wounds (2; 2.6%); and explosion (1; 1.3%). The circumstances of death and cause of fracture(s) were 'unascertained' in four cases (5.1%). Postmortem artefact accounted for two cases (2.6%).


Quality of life among survivors of adolescent and young adult cancer in Canada: A Young Adults With Cancer in Their Prime (YACPRIME) study.
The quality of life (QoL) among survivors of adolescent and young adult (AYA) cancer may be compromised compared with that in the general population. In this study, the authors: 1) assessed for differences in QoL among a national study of AYA cancer survivors compared with the Canadian population and 2) explored the factors associated with poorer QoL in AYA cancer survivors. For the current research, data from the Young Adults With Cancer in Their Prime study were used. QoL was measured using physical and mental component scores from a 12-item short-form health status measure. A comparison group was derived from the Canadian Community Health Survey. AYAs (n = 195; 17.8% male; mean ± SD: 35.62 ± 6.89 years on study, 6.48 ± 5.73 years from treatment) were compared with a comparison sample (n = 665; 21.2% male). Among survivors, 31.8% reported poor physical health, and 49.7% reported poor mental health. Compared with the general population, AYAs had significantly lower physical health (F[1,818] = 52.80; P = .00) and mental health (F[1,818] = 83.54; P = .00), controlling for sex and age. Multivariable logistic regression analyses revealed that an annual income level <$40,000 (adjusted odds ratio [AOR], 8.32; 95% CI, 2.85-24.30), poor sleep quality (AOR, 1.19; 95% CI, 1.06-1.33), worse body image (AOR, 1.08; 95% CI, 1.02-1.14), and higher social support (AOR, 1.02; 95% CI, 1.00-1.05) were significantly associated with poor physical health. Poor sleep quality (AOR, 1.22; 95% CI, 1.08-1.38), body image (AOR, 1.06; 95% CI, 1.01-1.12), fear of cancer recurrence (AOR, 1.13; 95% CI, 1.06-1.21) were associated with poor mental health. The QoL of AYAs requires urgent attention. Sleep, body image, and social support may be important modifiable targets for intervention to improve their QoL.


Mouse brain contains a high molecular weight nonfunctional protein with antigenic homology to dihydrofolate reductase.
The cytosol of brain tissue from mature BDF1 mice contains very little dihydrofolate reductase activity but it does contain a high molecular weight nonfunctional protein which cross-reacts in a radioimmunoassay for the active enzyme. Liver cytosol contains less of this high molecular weight cross-reacting protein and more of the functional dihydrofolate reductase. The cytosol from kidney contains very little of the high molecular weight cross-reacting protein, 95% of the immunoreactive proteins being the functional form of dihydrofolate reductase. The modification of dihydrofolate reductase into a nonfunctional form may be an intrinsic property of some cells and this finding could explain the variability in measuring the activity of this enzyme in brain tissue of mature animals.


Skeletal hormones and the C/EBP and Runx transcription factors: interactions that integrate and redefine gene expression.
Systemic hormones and local growth factors have significant and often complex roles in normal tissue development, growth, remodeling, and repair. Early efforts in skeletal tissue attempted to define active panels of these agents and their direct effects on cell proliferation, matrix production, and secretion of other soluble mediators of differentiated cell function. Initial results resolved many of these questions and began to unveil functional interactions between specific hormones and growth factors. More recent evidence suggests that interactions between individual hormone systems also occur in less anticipated but probably not less meaningful ways. In some cases, these interactions may help to define a spectrum of effects on gene expression by focusing, refocusing, or integrating the activity of previously recognized transcription regulators. Other studies in isolated osteoblasts predict that certain steroid hormones have distinctive effects on specific transcription factors with important roles in bone growth and repair. In this review, we focus on studies that define functional and physical interactions between molecular mediators of hormone activity that could directly effect skeletal growth factor biology.


Higher food intake and appreciation with a new food delivery system in a Belgian hospital. Meals on Wheels, a bedside meal approach: a prospective cohort trial.
A new system of meal distribution called Meals on Wheels, allowing food ordering at mealtime and providing guidance by trained nutritional assistants, might show benefit in offering nutritional support. This study investigates whether Meals on Wheels improves total food intake per day and yielded improved appreciation of food quality and increased access to food and mealtimes. In a prospective cohort trial where control and intervention groups were taken from all patients hospitalized at the respiratory disease department, age, sex, BMI, admission weight, height, reason for admission and discharge weight were noted, as was food intake, supplements, waste per meal and daily total. For food appreciation the questionnaire developed by Naithani et al. was used. The study included 83 patients in the control group and 106 patients in the Meals on Wheels group. Mean total daily food intake was 236 g higher in patients in the Meals on Wheels than in controls. There was higher intake of oral nutritional supplements in the Meals on Wheels group compared to controls, resulting in significantly less oral nutritional supplements wasted. There was also significantly less waste in the Meals on Wheels group. For food access and appreciation, patients appreciated Meals on Wheels more than the old system in terms of choice, hunger, food quality and organization. Meals on Wheels resulted in higher food intake during each meal, less waste and better use of oral nutritional supplements. Patients appreciated Meals on Wheels more than the old system in terms of choice, hunger, food quality and organization.


Social Interactions of Dat-Het Epi-Genotypes Differing for Maternal Origins: The Development of a New Preclinical Model of Socio-Sexual Apathy.
Social interaction is essential for life but is impaired in many psychiatric disorders. We presently focus on rats with a truncated allele for dopamine transporter (DAT). Since heterozygous individuals possess only one non-mutant allele, epigenetic interactions may unmask latent genetic predispositions. Homogeneous "maternal" heterozygous offspring (termed MAT-HET) were born from dopamine-transporter knocked-out (DAT-KO) male rats and wild-type (WT) mothers; "mixed" heterozygous offspring (termed MIX-HET) were born from both DAT-heterozygous parents. Their social behavior was assessed by: partner-preference (PPT), social-preference (SPT) and elicited-preference (EPT) tests. During the PPT, focal MIX-HET and MAT-HET males had a choice between two WT females, one in estrous and the other not. In the SPT, they met as stimulus either a MIX-HET or a WT male. In the EPT, the preference of focal male WT rats towards either a MIX- or a MAT-HET stimulus was tested. MIX-HET focal males showed an abnormal behavior, seeming not interested in socializing either with a female in estrous or with another male if MIX-HET. Focal MAT-HET males, instead, were very attracted by the female in estrous, but totally ignored the MIX-HET male. We assessed the expression of noradrenaline transporter (NET) in prefrontal cortex, hippocampus and hypothalamus, finding differences between the two offspring. MIX-HETs' hypothalamus and hippocampus showed less NET than MAT-HETs, while the latter, in turn, showed higher NET than WTs. These behavioral differences between heterozygous groups may be attributed to different maternal cares received. Results allow preclinical understanding of epigenetic factors involved in social-behavior abnormalities, typical of many psychiatric disorders.


Prediction value of procalcitonin combining CURB-65 for 90-day mortality in community-acquired pneumonia.
Background: Due to its high mortality rate, immediate and reliable severity assessment and accurate prediction of prognosis at hospital admission is critical for the management of community-acquired pneumonia (CAP) patients.Methods: Consecutive patients with primary diagnosis of CAP and hospitalized at our hospital from January 2013 to December 2015 were screened for this retrospective study. Demographic information, clinical and laboratory examination, severity model scoring, and 90-day outcomes were studied. Area under the curve (AUC) of receiver operating characteristic curve (ROC) was analyzed to compare the predictive value of different prognostic scoring methods.Results: 2099 CAP patients with a median age of 60 (IQR 44.0-73.0) years-old were included in this study. Median length of stay was 10 days (IQR 8.0-13.0). The all-cause 90-day mortality was found in 2.19% (46/2099) of all patients. PCT was identified as an independent predictor for the prognosis of CAP patients. CURB-65 in combination with PCT outperformed other predictive methods in 90-day mortality with the optimal AUC of 0.900 and Youden's Index of 0.706.Conclusions: PCT is a good marker for the assessment of severity and 90-day mortality of CAP patients. The combination of PCT and CURB-65 was more accurate than other prognostic models in predicting 90-day mortality.


Respiratory outcomes in high-risk children 7 to 10 years after prophylaxis with respiratory syncytial virus immune globulin.
Respiratory syncytial virus infections have been implicated in the development of asthma. We evaluated the long-term effects of respiratory syncytial virus immune globulin, an effective prophylactic agent for the prevention of these infections in children, on respiratory and allergic outcomes in children at high risk of chronic airway disease. Thirteen children at high risk of respiratory disease (mean [+/-SD] age, 8.6 +/- 1.1 years) were evaluated using pulmonary function and allergy skin testing 7 to 10 years after they had received prophylaxis with respiratory syncytial virus immune globulin. For comparison, 26 high-risk control children (mean age, 8.5 +/- 0.9 years) were also evaluated. Health outcomes data were collected from all subjects. The children were matched for age and gestational age. There were more boys, and a lesser frequency of a lower respiratory tract infection with respiratory syncytial virus (P <0.001) in the group that had been treated prophylactically than in the controls. The ratio of the forced expiratory volume in 1 second to forced vital capacity was significantly better in children who had received immune globulin (median, 0.88; interquartile range, 0.81 to 0.91) than in the controls (median, 0.76; interquartile range, 0.67 to 0.86; P = 0.02). Children were also less atopic (2 of 13) in the respiratory syncytial virus immune globulin group than in the control group (13 of 26, P <0.04) and were less likely to have missed school (P = 0.006) or have had an asthma attack (P = 0.03). The results suggest that prophylaxis of respiratory syncytial virus infections in infancy may have long-term effects on respiratory and immunologic parameters relevant to the development of asthma. Larger-scale studies are needed.


Ultrasound-determined fetal subcutaneous tissue thickness for a birthweight prediction model.
The aim of this study was to explore a birthweight prediction model using ultrasound determined tissue thickness (SCTT) parameters. We measured routine ultrasonographic biometric parameters and in addition, fetal SCTT in 201 healthy singleton pregnancies. Mid-arm fat and lean mass, mid-thigh fat and lean mass, subscapular fat mass and abdominal fat mass (AFM) were measured in order to calculate a birthweight prediction model. Ultrasound measurements were analyzed using an 'anovarepeated measures model'. The growth rate (beta-slope) of the selected parameters was computed and the correlation coefficient with the birthweight and the Kendall rank correlation tau, were calculated. From the ultrasound determined SCTT parameters, only abdominal circumference (AC), AFM, and MTLM showed a statistically significant trend. The beta-slope of mid-thigh lean mass was excluded since it exhibited significant correlation with the beta-slope of AFM. The final regression model could be calculated as: birthweight (gr.) = intercept +alpha(1)(AFM beta-slope) + alpha(2)(AC beta-slope), where alpha(1), alpha(2) represent regression coefficients. We provide a graphical birthweight prediction model for clinical practice using conventional and specific ultrasound measurements of fetal subcutaneous tissue thickness. This model is based upon an overall analysis of the ultrasound estimated body components.


Diuretic, natriuretic and potassium-sparing effect of nothofagin isolated from Leandra dasytricha (A. Gray) Cogn. leaves in normotensive and hypertensive rats.
Active constituents from natural origin have long been used for the treatment of patients suffering from cardiovascular and renal diseases. This study therefore aimed to investigate the diuretic and natriuretic properties of nothofagin, a dihydrochalcone isolated from Leandra dasytricha (A. Gray) Cogn. leaves in normotensive and hypertensive rats. Male Wistar normotensive rats were orally treated with vehicle (1 ml/kg); hydrochlorothiazide (HCTZ; 25 mg/kg); ethyl acetate fraction from L. dasytricha (EALD; 3-30 mg/kg) and nothofagin (NOT; 0.3-3 mg/kg). Spontaneously hypertensive rats (SHR) received NOT (1 mg/kg), HCTZ (25 mg/kg) or vehicle. The cumulative diuretic index, urinary electrolytes excretion (Na+ and K+), pH, density and conductivity were measured at the end of the experiment (after 8 h). A7r5 and L929 cell lines were used to measure cell viability after exposure to NOT. Nitric oxide generation was quantified in A7r5 cell supernatant, and DPPH assay was used for evaluating the antioxidant properties of NOT. The urinary volume of normotensive rats were increased after the treatment with EALD, without any changes in Na+ or K+ excretion. NOT was able to induce diuresis and natriuresis, but not kaliuresis, in both normotensive and hypertensive rats. The reduction in prostanoids generation through cyclooxygenase inhibition, as well as the muscarinic receptor antagonism, fully avoided NOT-induced increases in diuretic index. NOT, which did not interfere with L929 or A7r5 cell viability, was able to stimulate nitric oxide generation in A7r5 cell, besides showing an antioxidant effect in scavenging the free-radical DPPH. Taken together, our study shows, for the first time, the diuretic, natriuretic and potassium-sparing effect of nothofagin in rats, which was associated with prostanoids generation, muscarinic receptor activation and antioxidant properties.


Longitudinal variance of Donor-Derived Cell-Free DNA (dd-cfDNA) in Stable Kidney Transplant (KTx) patients are influenced by donor/recipient variables.
The longitudinal time-course of dd-cfDNA after kidney transplant (KTx) is not well-described. The cut off values of dd-cfDNA in KTx derive from biopsy-coupled single measurements. Meaningful interpretation necessitates understanding of: (1) time variance of dd-cfDNA levels post-KTx, (2) factors determining biologic variability, and (3) relationship to donor and recipient characteristics. We hypothesized that an understanding of the aforementioned factors would better inform clinical decision-making using dd-cfDNA. One hundred and twenty five KTx patients with dd-cfDNA obtained longitudinally were included. Univariate analyses were directed at inter-patient variability and intra-patient inter-occasion variability of dd-cfDNA. Multivariate linear regression was used in analyses accounting for repeat measures. At 1-month post KTx median dd-cfDNA: (1) were higher in repeat KTx (.57%, P < .001), and dual KTx (1.10%, P = ns) versus a first KTx (.31%); (2) showed a significant difference in donor after cardiac death (DCD [.45%]) versus living related (LRD [.27%]) donors (P = .036). Longitudinal (1-3 months) dd-cfDNA measurements showed a significant downtrend for all donor types. Panel-reactive antibodies (PRA) were positively correlated with dd-cfDNA. Repeat Tx, dual Tx, DCD, and PRA are associated with a higher dd-cfDNA. Incorporation of donor/recipient variables and time down post transplant is material for rational interpretation of dd-cfDNA.


Serratus anterior plane versus paravertebral nerve blocks for postoperative analgesia after non-mastectomy breast surgery: a randomized controlled non-inferiority trial.
Paravertebral and serratus plane blocks are both used to treat pain following breast surgery. However, it remains unknown if the newer serratus block provides comparable analgesia to the decades-old paravertebral technique. Subjects undergoing unilateral or bilateral non-mastectomy breast surgery were randomized to a single-injection serratus or paravertebral block in a subject-masked fashion (ropivacaine 0.5%; 20 mL unilateral; 16 mL/side bilateral). We hypothesized that (1) analgesia would be non-inferior in the recovery room with serratus blocks (measurement: Numeric Rating Scale), and (2) opioid consumption would be non-inferior with serratus blocks in the operating and recovery rooms. In order to claim that serratus blocks are non-inferior to paravertebral blocks, both hypotheses must be at least non-inferior. Within the recovery room, pain scores for participants with serratus blocks (n=49) had a median (IQR) of 4.0 (0-5.5) vs 0 (0-3.0) for those with paravertebral blocks (n=51): 0.95% CI -3.00 to -0.00; p=0.001. However, the difference in morphine equivalents did not reach statistical significance for superiority with the serratus group consuming 14 mg (10-19) vs 10 mg (10-16) for the paravertebral group: 95% CI -4.50 to 0.00, p=0.123. Since the 95% CI lower limit of -4.5 was less than our prespecified margin of -2.0, we failed to conclude non-inferiority of the serratus block with regard to opioid consumption. Serratus blocks provided inferior analgesia compared with paravertebral blocks. Without a dramatic improvement in safety profile for serratus blocks, it appears that paravertebral blocks are superior to serratus blocks for postoperative analgesia after non-mastectomy breast surgery. NCT03860974.


Pharmacokinetic Comparison of Chitosan-Derived and Biofermentation-Derived Glucosamine in Nutritional Supplement for Bone Health.
Glucosamine and chondroitin sulfate have been used as nutritional supplementation for joint tissues and osteoarthritis (OA). Biofermented glucosamine is of great interest in the supplement industry as an alternative source of glucosamine. The purpose of this study is to compare the pharmacokinetics of chitosan-derived glucosamine and biofermentation-derived glucosamine as nutritional supplementation. In a randomized, double-blind and cross-over study design, we recruited subjects of healthy men and women. The pharmacokinetics of glucosamine were examined after a single dose of glucosamine sulfate 2KCl (1500 mg) with two different sources of glucosamine (chitosan-derived glucosamine and biofermentation-derived glucosamine) to male and female subjects fitted with intravenous (iv) catheters for repeated blood sampling up to 8 h. According to plasma concentration-time curve of glucosamine after an oral administration of 1500 mg of glucosamine sulfate 2KCl, AUC0-8h and AUC0-∞ values of glucosamine following oral administration of chitosan-derived and biofermentation-derived glucosamine formulations were within the bioequivalence criteria (90% CI of ratios are within 0.8-1.25). The mean Cmax ratios for these two formulations (90% CI of 0.892-1.342) did not meet bioequivalence criteria due to high within-subject variability. There were no statistically significant effects of sequence, period, origin of glucosamine on pharmacokinetic parameters of glucosamine such as AUC0-8h, AUC0-∞, Cmax. Our findings suggest that biofermentation-derived glucosamine could be a sustainable source of raw materials for glucosamine supplement.


High-resolution cryo-EM structure of photosystem II reveals damage from high-dose electron beams.
Photosystem II (PSII) plays a key role in water-splitting and oxygen evolution. X-ray crystallography has revealed its atomic structure and some intermediate structures. However, these structures are in the crystalline state and its final state structure has not been solved. Here we analyzed the structure of PSII in solution at 1.95 Å resolution by single-particle cryo-electron microscopy (cryo-EM). The structure obtained is similar to the crystal structure, but a PsbY subunit was visible in the cryo-EM structure, indicating that it represents its physiological state more closely. Electron beam damage was observed at a high-dose in the regions that were easily affected by redox states, and reducing the beam dosage by reducing frames from 50 to 2 yielded a similar resolution but reduced the damage remarkably. This study will serve as a good indicator for determining damage-free cryo-EM structures of not only PSII but also all biological samples, especially redox-active metalloproteins.


Frontal Sinus Repair Using Polymethyl Methacrylate After Bifrontal Craniotomy.
In many cases in which bifrontal craniotomy is performed, the frontal sinus is opened, and postoperative complications occur. Various methods to close the frontal sinus have been reported. However, all these methods require skill to perform and take time. The aim of this study was to report results obtained with closure of the frontal sinus using polymethyl methacrylate, which is a simpler method. From December 2008 to June 2018, 122 patients with a ruptured anterior communicating aneurysm were transported to the authors' facility, and 52 patients consecutively underwent bifrontal craniotomy with opening of the frontal sinuses, which were then filled with polymethyl methacrylate. Mean follow-up period was 21.8 months; the longest follow-up was 116 months. No cerebrospinal fluid leakage or other complications were observed. The method of filling the frontal sinus with polymethyl methacrylate reported in this study was safe and had no complications. This method should be considered as a method of frontal sinus closure.


Psychological distress and mental health literacy in university students.
This study aimed to assess university students' levels of psychological distress and mental health literacy. This descriptive and cross-sectional study using self-assessment instruments with 417 volunteer university students was conducted in the fall semester of the 2019-2020 academic year. More than half of the participants had diagnosable psychological distress in terms of mental illness. The participants had a low level of mental health literacy, females and people with a mental illness had higher mental health literacy scores. The results indicated that the scales could be used to develop interventions to assist Turkish students' transition to healthy adulthood. Determining individuals' psychological distress allows early detection of mental problems.


Is Coronoidectomy Superior to Coronoidotomy for Improving Maximum Incisal Opening in Patients With Oral Submucous Fibrosis?
In the management protocol of the oral submucous fibrosis (OSMF), multiple studies have advocated that an additional step of coronoidectomy or coronoidotomy helps achieve better and more consistent postoperative results. But, there are no studies that validate if one has an advantage over the other. This study aimed to compare the outcome of the coronoidectomy with coronoidotomy for OSMF cases. A randomized controlled trial was designed in patients with OSMF requiring operative intervention. Patients were randomized into 2 groups based on a computer-generated randomization table: group I coronoidectomy and group II coronoidotomy. The primary predictor variables were the 2 different adjunct surgical steps - coronoidectomy and coronoidotomy. The primary outcome variable was the postoperative maximum incisal opening (MIO). Secondary outcome variables were duration of surgery and blood loss. The study sample was composed of 32 patients (16 each group). Both the groups were comparable in terms of demographic profile. The MIO after surgery was comparable throughout the study between the groups. Repeated measures analysis of variance for comparison of mouth opening within the group showed that there was a significant improvement in postoperative MIO in both the groups (group I P value <.001 and group II P value 0.004). A statistically significant difference was found in the duration of the surgery (3.5 ± 0.73 hours vs 2.06 ± 0.87 hours) and blood loss (393 .75 ± 278.6 mL vs 90.62 ± 58.36 mL) with the shorter time and less blood loss in coronoidotomy compared with coronoidectomy. All these cases were followed for 1 year. Coronoidotomy as an adjunctive treatment in OSMF provides comparable treatment outcome in terms of MIO compared with coronoidectomy with the added advantage of shorter operating time and less blood loss.


[Pathomorphological assessment of the course of tuberculosis in Tashkent].
Fatal cases of tuberculosis were analyzed in 3 groups: 1) Tashkent citizens registered in tuberculosis dispensaries (83.39%); 2) those unregistered there (11.1%); 3) Tashkent migrants (5.6%). Main clinical entities of tuberculosis, such as fibrocavenous tuberculosis (88.9%), infiltrative tuberculosis (3.7%), and disseminated tuberculosis (3.7%) were found to be causes of death.


Idebenone induces apoptotic cell death in the human dopaminergic neuroblastoma SHSY-5Y cells.
Idebenone is a coenzyme Q10 analog and an antioxidant that has been used clinically to treat Friedreich Ataxia. Being an antioxidant, idebenone could have potential therapeutic potential to treat other neurodegenerative diseases such as Parkinson's disease in which oxidative stress plays a role in their pathogenesis. But whether idebenone can be used to treat Parkinson's disease has not been evaluated. In this study, we found that exposure of the dopaminergic neuroblastoma SHSY-5Y cells to 1-10 μM idebenone for 72 h had no effect on the cell viability revealed by trypan blue exclusion assay and MTT assay. However, cells exposed to 25 μM or higher concentrations of idebenone showed extensive trypan blue-positive staining and significant reduction in cell viability revealed by MTT assay indicating that most of the cells were no longer viable. Idebenone-induced cell death was characterized by genomic DNA fragmentation and accumulation of cytochrome c in the cytosol indicating that the death was apoptotic in nature. In addition, idebenone induced an increase in the total RNA of the pro-apoptosis protein BAX, it also increased the caspase-3 activity in the cell lysates when compared with the untreated control cells or cells exposed to 10 μM or lower concentrations of idebenone. The detrimental effect of idebenone was attenuated by glutathione, an antioxidant, suggesting that oxidative stress contributed to the idebenone-induced cell death. In conclusion, our results suggest that antioxidant idebenone induced apoptosis when used in high concentrations.


Quantification of ghrelin and des-acyl ghrelin in human plasma by using cubic-selected reaction-monitoring LCMS.
Ghrelin is a peptide hormone generally measured in plasma by immunoassays. LCMS/MS was investigated as an alternative method in particular for the quantification of the two forms of the peptide with improved selectivity. A LCMS assay using a cubic-selected reaction-monitoring (LCSRM(3)/MS) mode was developed for the quantification of ghrelin and des-acyl ghrelin in human plasma. The LCSRM(3)/MS method was found to be linear from 50-75 to 2500 pg/ml for the ghrelins using a 0.5-ml plasma sample. The accuracies and precisions at LOQ for des-acyl ghrelin (50 pg/ml) and ghrelin (75 pg/ml) were found to be better than 91 and 2%, respectively. Blood and plasma stabilization was found to be essential for good assay performance. Compared to the LCSRM/MS method the addition of an additional MS step did significantly improve the selectivity and therefore the sensitivity. The LCSRM(3)/MS method could be successfully applied for the quantification of ghrelin and des-acyl ghrelin in human plasma samples.


Autoantibodies to GAD, IA-2 and insulin in ICA-positive first-degree relatives of children with type 1 diabetes: a comparison between parents and siblings.
Islet cell antibodies (ICA) represent a heterogenous group of autoantibodies to diabetes-associated antigens, including glutamic acid decarboxylase (GAD) and the IA-2 protein. The objectives of the present study were to compare the prevalence of autoantibodies to known biochemically characterized autoantigens between ICA-positive non-diabetic parents and siblings of children with type 1 diabetes and to evaluate how such antibodies explain ICA reactivity. The presence and levels of GAD antibodies (GADA), IA-2 antibodies (IA-2A) and insulin autoantibodies (IAA) were analyzed in the sera of 184 ICA-positive first-degree relatives (79 parents and 105 siblings). The prevalences of GADA (61.9% in siblings vs 32.9% in parents), IA-2A (55.2% vs 15.2%) and IAA (41.0% vs 0%) were increased among ICA-positive siblings relative to ICA-positive parents (p<0.001). The siblings had higher ICA titers (p<0.001) than the parents but tended to have lower GADA levels (p=0.12). IA-2A levels did not differ between the two groups. IA-2A levels explained a higher proportion of the ICA reactivity in the siblings than in the parents (44% vs 12%, p=0.004), and GADA levels had the same tendency (27% vs 10%, p=0.11). In a multiple regression analysis, GADA and IA-2A were found to explain together 16% of the ICA reactivity in parents and 49% in siblings (p=0.003 for the difference). These results indicate that the increased frequency of additional diabetes-associated autoantibodies in ICA-positive siblings when compared to their ICA-positive parents may reflect the increased risk of progression to clinical type 1 diabetes previously reported in young ICA-positive relatives. We conclude that ICA immunofluorescence is not only due to GADA and IA-2A, but there are other additional antigens contributing to the ICA reactivity. Antibodies to such antigens appear to be more common among adults than in children.


Thyroid carcinoma metastasized to the sternum: resection of the sternum and reconstruction with acrylic resin.
Recently we resected and reconstructed the sternum in two female patients with thyroid carcinoma metastatic to the sternum. Histological diagnosis of the metastatic lesion was follicular carcinoma in the first case and papillary carcinoma in the second case. Reconstruction of the sternum was accomplished by using an acrylic resin plate alone in the first case and an acrylic resin plate sandwiched between layers of Marlex mesh (Marlex sandwich procedure) in the second case. In the first case the acrylic resin plate was directly fixed to the ribs with metallic wires, most of which snapped off later, and a small amount of exudate accumulated around the plate for a short period of time. In contrast, the postoperative course in the second case was uneventful and the Marlex sandwich procedure seemed superior in the chest wall reconstruction following resection of the sternum.


Therapeutic inhibition of keratinocyte TRPV3 sensory channel by local anesthetic dyclonine.
The multimodal sensory channel transient receptor potential vanilloid-3 (TRPV3) is expressed in epidermal keratinocytes and implicated in chronic pruritus, allergy, and inflammation-related skin disorders. Gain-of-function mutations of TRPV3 cause hair growth disorders in mice and Olmsted syndrome in humans. Nevertheless, whether and how TRPV3 could be therapeutically targeted remains to be elucidated. We here report that mouse and human TRPV3 channel is targeted by the clinical medication dyclonine that exerts a potent inhibitory effect. Accordingly, dyclonine rescued cell death caused by gain-of-function TRPV3 mutations and suppressed pruritus symptoms in vivo in mouse model. At the single-channel level, dyclonine inhibited TRPV3 open probability but not the unitary conductance. By molecular simulations and mutagenesis, we further uncovered key residues in TRPV3 pore region that could toggle the inhibitory efficiency of dyclonine. The functional and mechanistic insights obtained on dyclonine-TRPV3 interaction will help to conceive therapeutics for skin inflammation.


Nasal nitric oxide and other diagnostic procedures in seasonal allergic rhinitis: elderly vs juvenile patients.
The type of allergy and other diagnostic procedures were analyzed in a group of elderly seasonal allergic rhinitis (SAR) patients and were compared with young SAR people and a control group. Study group consisted of 248 patients with mean age 64.1 ± 4.2 years. They were compared with 289 young SAR people (mean age, 23.7 ± 5.8 years) and with a control group. Allergic sensitization was assessed using the skin prick test (SPT) and specific immunoglobulin (Ig) E measurements. The SAR symptoms were monitored using visual graphic scale. Measurement of nasal fractional exhaled nitric oxide (FeNO) level obtained with handheld chemiluminescence analyzer during and after pollen season was compared with control groups. In the elderly patient group, a prevalence of allergy to grass, rye, and mugwort was noticed. In 134 (54%) patients, SPT results to grass were positive; and the specific IgE level was elevated (mean value, 22.8 ± 13.1 IU/mL), whereas in 83 (33%) subjects, positive SPT results to mugwort and elevated serum specific IgE concentration (mean value, 31.9 ± 9.1 IU/mL) were observed. During the pollen season, a significantly higher FeNO level was noted in both young and elderly SAR patients in comparison with their respective control groups. In young patients, the mean FeNO level was 61.8 ± 17.2 parts per billion, whereas in the elderly group, it was 58.2 ± 11.8 parts per billion. SAR in elderly patients is a problem that cannot be neglected. The natural course of SAR in the elderly is similar to other age groups. However, hypersensitivity to different pollens in the analyzed group needs further study.


Clostridium septicum growth from a total knee arthroplasty associated with intestinal malignancy: a case report.
Previous reports of infection with Clostridium septicum have identified an unexpected association with concurrent malignancy. The reported rate of associated malignancy has been found to be as high as 81 percent. The purpose of this case report was to present a case of a total knee arthroplasty infected with C. septicum and the subsequent finding of an occult colonic malignancy. A 74 year-old man underwent uneventful bilateral total knee arthroplasties. Two weeks post-operatively, he presented with acute swelling, redness and pain of the left knee. Aspiration of the knee was sent for cell count and culture. The cell count demonstrated 39,000 white blood cells per cubic millimeter with 71% of white blood cells identified as neutrophils. Synovial fluid cultures identified the presence of C. septicum, Enterobacter and coagulase negative Staphylococcus. After urgent irrigation and debridement and polyethylene exchange of the affected knee, the patient was placed on intravenous Penicillin G for a period of six weeks. Two weeks into his course of antibiotics, the patient developed hematochezia and was found to have an obstructive colonic malignancy. The patient underwent hemi-colectomy and has since made a complete recovery of both his malignancy and total knee arthroplasty infection. Recognition of the association between C. septicum and malignancy is especially important considering the large predicted increase in total joint arthroplasty procedures over the coming decades. In addition to the standard treatment for infection after total joint arthroplasty, identification of Clostridium septicum should initiate a search for associated occult malignancy.


How does air quality affect residents' life satisfaction? Evidence based on multiperiod follow-up survey data of 122 cities in China.
Since entering the new era, China's socialist contradiction has been transformed into the contradiction between the people's growing need for a better life and the unbalanced and inadequate development. How to improve the quality of people's life through the improvement of air quality has become an important content restricting social development and a key problem to be solved. Based on the life satisfaction (LS) method, this study takes air quality into the individual utility function, and through matching China Health and Retirement Longitudinal Study (CHARLS), two phases of microindividual tracking data with 122 urban environmental quality data innovatively investigate the impact of air quality on residents' LS and its income substitution effect. The results show that air quality significantly reduces residents' LS, among which, different air pollutants and comprehensive air quality AQI have significant negative effects. And PM10 has the highest marginal effect on different LS evaluation, SO2 has the smallest marginal effect, and AQI marginal effect is close to PM10. In terms of group heterogeneity, NO2 and SO2 have group influence differences in age group, regional economic group, gender group, and family per capita income group. But PM10 and AQI do not show group influence heterogeneity, and air quality has significant negative effect on LS of different groups. In addition, the interaction between air quality and income level shows that air quality strengthens the difference of residents' LS caused by income level difference. According to the equilibrium condition of residents' individual utility function, the improvement of air quality by 1% is equivalent to the improvement of residents' LS by 23.4402% of income. Firstly, air quality has an important impact on residents' LS, and different air pollutants have different effects. Secondly, the impact of air quality on LS of different groups is heterogeneous and mainly diversified in age group, regional economic group, gender group, and family per capital income group. Finally, there is substitution effect between air quality and regional GDP growth and household income, which affects residents' LS. Thirdly, the conclusion shows that the improvement of air quality is difficult to be replaced by other ways. Good air quality can not only directly improve residents' LS, but also has economic effect.


Impact of communication modality on caregiver ratings for deaf and hard of hearing children.
Literature on children who are deaf or hard of hearing (DHH) suggests overall increased rates of difficulties in emotional/behavioral and adaptive functioning. However, limitations of this literature include the failure to integrate issues unique to the experience of children who are DHH, such as home and school communication modalities and the consistency of modalities across settings. This study examined de-identified data from a clinical database. Data included caregiver ratings of emotional/behavioral and adaptive functioning in a diverse sample of clinically referred children who are DHH (N = 177). Caregivers also reported home and school communication modalities (e.g., match, partial match, different modalities). We examined mean score differences between our sample and normative samples and compared functioning across subgroups of children with various home-school communication modality combinations. Consistent with the literature, we found overall increased rates of emotional/behavioral and adaptive functioning concerns on parent rating scales. Emotional/behavioral concerns did not differ among children with spoken language match, sign language match, or partial match communication modalities combinations. Within adaptive functioning, communication and functional academics were significantly lower among children with partial match home-school communication modalities. Adaptive functioning did not differ between spoken language match and sign language match groups. Our findings suggest possible benefits to adaptive functioning among children who are DHH when home and school communication modalities match, regardless of which modality is used.


Date of marine annulus formation in Atlantic salmon (Salmo salar) and implications for retrospective growth analyses using scales.
Fish scales have increasingly been used to quantify annual and seasonal growth trends and in efforts to relate growth to environmental conditions. Understanding the timing of formation of an annulus (a group of narrowly spaced circuli) is critical when assessing the influence of marine ecosystem conditions on seasonal growth patterns of Atlantic salmon, yet the literature does not provide consistent answers regarding the timing or drivers of marine annulus formation. This study demonstrates a novel method for estimating stock-specific annulus formation timing based on marked individuals with known emigration and return/recovery dates. An equation was applied to estimate the date of annulus completion for Atlantic salmon (Salmo salar) using known dates, number of circuli after the most recent annulus and marine circulus deposition rate. Five marine circulus deposition rate scenarios were tested, some of which accounted for individual, seasonal and age-related variability and others which use previously published marine circulus deposition rates. Based on these results, an argument is presented to reconsider the practice of assigning annulus formation dates to winter solstice in favour of dates estimated by a scenario that accounts for individual, seasonal and age-related variation in circulus deposition. This scenario suggests that annulus formation occurs between mid-February and late March. In this case, the annulus would be formed during the coldest part of the year in the primary overwintering area for North American Atlantic salmon.


Geographical Distribution and Pattern of Pesticides in Danish Drinking Water 2002-2018: Reducing Data Complexity.
Pesticides are a large and heterogenous group of chemicals with a complex geographic distribution in the environment. The purpose of this study was to explore the geographic distribution of pesticides in Danish drinking water and identify potential patterns in the grouping of pesticides. Our data included 899,169 analyses of 167 pesticides and metabolites, of which 55 were identified above the detection limit. Pesticide patterns were defined by (1) pesticide groups based on chemical structure and pesticide-metabolite relations and (2) an exploratory factor analysis identifying underlying patterns of related pesticides within waterworks. The geographic distribution was evaluated by mapping the pesticide categories for groups and factor components, namely those detected, quantified, above quality standards, and not analysed. We identified five and seven factor components for the periods 2002-2011 and 2012-2018, respectively. In total, 16 pesticide groups were identified, of which six were representative in space and time with regards to the number of waterworks and analyses, namely benzothiazinone, benzonitriles, organophosphates, phenoxy herbicides, triazines, and triazinones. Pesticide mapping identified areas where multiple pesticides were detected, indicating areas with a higher pesticide burden. The results contribute to a better understanding of the pesticide pattern in Danish drinking water and may contribute to exposure assessments for future epidemiological studies.


Saccharides in atmospheric particulate and sedimentary organic matter: Status overview and future perspectives.
Saccharides are omnipresent compounds in terrestrial and marine ecosystems. Since the 2000s, their role in environmental and geochemical studies has significantly increased, but only anhydrosaccharides (mainly levoglucosan) have been reviewed. Here we present the wider knowledge about saccharides in organic matter of aerosols, bottom sediments, soils, dust, and sedimentary rocks. The main purpose here is to characterize the possible sources of saccharides, as well as sacharol formation, seasonal variability, and the possible applications in environmental and paleoenvironmental interpretations. Different saccharide sources were designated, including biomass burning, and particulate matter such as pollen, spores, lichen, and fungi, as well as polysaccharide decomposition as possible inputs of monosaccharides. The main focus was on the most common saccharides encountered in environmental samples and sedimentary rocks. These are the mono- and disaccharides glucose, fructose, sucrose, and trehalose, and sacharols arabitol and mannitol. The anhydrosaccharides levoglucosan, mannosan, and galactosan were evaluated as ancient wildfire indicators and industrialization tracers found in lacustrine sediments starting from Pleistocene to contemporary deposits. However, other anhydrosaccharides like xylosan and arabinosan were also found as products of fossil wood burning. These anhydrosaccharides have the potential to be further tracers of hemicellulose burning. Additional recommendations are proposed for future research, including environmental and paleoenvironmental topics that need to be addressed.


The value of lymphadenectomy in surgical resection of perihilar cholangiocarcinoma: a systematic review and meta-analysis.
Surgical resection is the only potentially curative treatment for patients with resectable perihilar cholangiocarcinoma (PHC). There is still no consensus on the value of lymphadenectomy despite evidence indicating lymph node (LN) status is an important prognostic indicator for postoperative long-term survival. We sought to perform a meta-analysis to summarize the current evidence on the value of lymphadenectomy among patients undergoing surgery for PHC. The PubMed (OvidSP), Embase and Cochrane Library were systematically searched for studies published before July 2020 that reported on lymphadenectomy at the time of surgery for PHC after curative surgery. 7748 patients from 28 studies were included in the meta-analysis. No survival benefit was identified with increased number of LN resected (all P > 0.05). Meanwhile, overall LN status was an important prognostic factor. Patients with lymph node metastasis had a pooled estimate hazard ratio of death that was over two-fold higher than patients without lymph node metastasis (HR 2.07, 95% CI 1.65-2.59, P < 0.001). The examination of 5 LNs on histology was associated with better staging of lymph node status and stratification of patients into positive or negative LN groups. While the extent of LN dissection was not associated with a survival benefit, examination of more than 5 LNs better staged patients into positive or negative LN groups with a lower risk of nodal understaging.


Cytogenetic, cross-mating and molecular evidence of four cytological races of Anopheles crawfordi (Diptera: Culicidae) in Thailand and Cambodia.
Twenty-nine isolines of Anopheles crawfordi were established from wild-caught females collected from cow-baited traps in Thailand and Cambodia. Three types of X (X1, X2, X3) and four types of Y (Y1, Y2, Y3, and Y4) chromosomes were identified, according to differing amounts of extra heterochromatin. These sex chromosomes represent four metaphase karyotypes, i.e., Forms A (X1, X2, X3, Y1), B (X1, X2, X3, Y2), C (X2, Y3) and D (X2, Y4). Forms C and D are novel metaphase karyotypes confined to Thailand, whereas forms A and B appear to be common in both Thailand and Cambodia. Cross-mating experiments between the four karyotypic forms indicated genetic compatibility in yielding viable progenies and synaptic salivary gland polytene chromosomes. The results suggest that the forms are conspecific and A. crawfordi comprises four cytological races, which is further supported by very low intraspecific variation (mean genetic distance=0.000-0.018) of the nucleotide sequences in ribosomal DNA (ITS2) and mitochondrial DNA sequences (COI, COII).


Epidemiology and treatment of heart failure in Spain: the HF-PATHWAYS study.
To describe the epidemiology and treatment of a large contemporary cohort of patients with heart failure (HF). Observational, retrospective, population-based study using the BIG-PAC database, which includes people aged ≥ 18 years seeking care for HF between 2017 and 2019. The main variables were the prevalence/annual incidence rate, comorbidities, clinical variables, and medication administered. We identified 19 762 patients with HF from a total of 1 189 003 persons seeking medical attention from 2017 to 2019 (2019: mean age, 78.3 years; 53.0% men). Distribution by type of left ventricular ejection fraction (LVEF) was as follows: 51.7% reduced, 40.2% preserved, and 8.1% mid-range. In 2019, the prevalence was 1.89% (95%CI, 1.70-2.08), with an incidence rate of 2.78 new cases per 1000 persons/y. No statistically significant differences were observed in prevalence and/or incidence from 2017 to 2019. Among patients with HF with reduced ejection fraction (HFrEF), 64% received beta-blockers, 80.5% angiotensin-converting enzyme inhibitor/angiotensin receptor blockers or sacubitril-valsartan, and 29.8% an aldosterone antagonist. In addition, from the diagnosis (baseline) to 24 months of follow-up, there was discreet treatment optimization, which was notable in the first 3 to 6 months. Epidemiological data on HF remained stable during the study period, with a lower prevalence than that reported in non-population-based studies. There is wide room for improvement in the optimization of medical treatment of HFrEF.


Evidence for cell surface control of macronuclear DNA synthesis in Stentor.
In cell grafts, Stentor macronuclei associated with separate regions of cell surface can be made asynchronous with regard to morphology and DNA synthesis even though they demonstrably share a common endoplasm. These results suggest a mechanism for nuclear differentiation within a single cytoplasmic compartment, based on cell surface differences.


Mercury, cadmium and lead concentrations in five species of freshwater fish from eastern England.
A total of 146 samples of five species of fish were examined between March and November 1986 in four sites from the Rivers Brett and Chelmer in eastern England. Variations in heavy metals concentrations between sites and species were related mainly to size differences of fish. Mercury levels in the flesh were higher than in the liver, while cadmium and lead levels were higher in the liver.


Functional reconstitution of carrier proteins by removal of detergent with a hydrophobic ion exchange column.
A method has been developed for the functional reconstitution of membrane proteins in phospholipid vesicles. This method is an extension of a previously published procedure (Ueno, M., Tanford, C. and Reynolds, A. (1984) Biochemistry 23, 3070-3076) for the formation of unilamellar vesicles from mixed micelles of egg phosphatidylcholine and dodecyl octaoxyethylene ether. Mixed micelles are formed from detergent-solubilized protein and egg-yolk phospholipid vesicles. These micelles are subjected to repeated passage through small columns filled with Amberlite XAD-2 beads. Several carrier proteins from the inner mitochondrial membrane have been reconstituted in this way; experimental data are shown for the aspartate/glutamate carrier and the ADP/ATP carrier. Certain parameters proved to be important for optimal efficiency of reconstitution: the ratio of detergent/phospholipid in the mixed micelles, the concentration of phospholipid during the hydrophobic chromatography, the ratio of phospholipid/protein, (d) the ratio of detergent/Amberlite XAD 2 beads, the number of column passages, and the type of detergent. After optimization of these parameters, phospholipid vesicles with a diameter of about 150 nm were obtained. The main advantage of this procedure, however, lies in the fact that high amounts of membrane protein can be incorporated into the phospholipid vesicles, i.e. up to 15% (w/w).


Regulated expression of proteins in yeast using the MAL61-62 promoter and a mating scheme to increase dynamic range.
The ability to express heterologous genes in yeast has become indispensable for many biological research techniques. Expression systems that can be regulated are particularly useful because they allow an experimenter to control the timing and levels of gene expression. Despite their many advantages, however, surprisingly few conditional expression systems are available for yeast. Moreover, of those that have been described, many are not ideal either because they have high background expression levels, low induced levels, or because they require restrictive growth conditions. Here we describe a conditional expression system that takes advantage of the yeast MAL62 promoter (MAL62p), which can be controlled by adding maltose or glucose to the growth medium to induce or repress transcription, respectively. In addition, we use a mating scheme to dramatically increase the dynamic range of expression levels possible. We show that MAL62p background activity can be effectively eliminated by maintaining expression constructs in a mal(-) yeast strain. High-level expression can be induced in diploids formed by mating the mal(-) strain with a MAL(+) strain. A similar mating scheme may be useful for other conditional expression systems as well. Among other uses, this approach should aid high throughput yeast two-hybrid assays, which rely on maintaining large libraries of expression strains, which are eventually mated to conduct assays for protein interactions. We demonstrate a two-hybrid system in which MAL62p is used in conjunction with the yeast GAL1 promoter to independently regulate expression of both hybrid proteins, and to allow detection of interactions involving toxic proteins.


Aromatase in axonal processes of early postnatal hypothalamic and limbic areas including the cingulate cortex.
It has been shown that sexual dimorphic morphology of certain hypothalamic and limbic areas underlie gender-specific sexual behavior and neuroendocrine mechanisms. The key role played by locally formed estrogen in these developmental events has been revealed during a critical perinatal period. In this study, we aimed to document the presence of estrogen-synthetase (aromatase)-immunoreactive elements in the involved limbic system and hypothalamus of the developing rat brain. On postnatal day 5, animals of both sexes were perfusion-fixed, and sections from the forebrain and hypothalamus were immunolabelled for aromatase using an antiserum that was generated against a 20 amino acid sequence of placental aromatase. Aromatase-immunoreactivity was present in neuronal perikarya and axonal processes in the following limbic structures: the central and medial nuclei of the amygdala, stria terminalis, bed nucleus of the stria terminalis (BNST), lateral septum, medial septum, diagonal band of Broca, lateral habenula and all areas of the limbic (cingulate) cortex. In the hypothalamus, the most robust labelling was observed in the medial preoptic area, periventricular regions, ventromedial and arcuate nuclei. The most striking feature of the immunostaining with this antiserum was its intracellular distribution. In contrast to the heavy perikaryal labelling that can be observed with most of the currently available aromatase antisera, in the present experiments, immunoperoxidase was predominantly localized to axons and axon terminals. All the regions with fiber staining corresponded to the projection fields of neuron populations that have previously been found to express perikaryal aromatase. Our results confirm the presence of aromatase-immunoreactivity in developing limbic and hypothalamic areas. The massive expression of aromatase in axonal processes raises the possibility that estrogen formed locally by aromatase may not only regulate the growth, pathfinding and target recognition of its host neuronal processes, but may also exert paracrine actions on structures in close proximity, including the target cells.


Synthesis, characterization and anti-diabetic therapeutic potential of a new benzyl acid-derivatized kojic acid vanadyl complex.
Vanadium complexes are potent hypoglycemic agents and of great potential for therapeutical treatment of diabetes. In the present work, a novel vanadium compound, bis ((5-hydroxy-4-oxo-4H-pyran-2-yl)methyl benzoatato) oxovanadium (IV) (BBOV) has been synthesized. Treatment of STZ-induced diabetic rats with BBOV restored the blood glucose to normal level and ameliorated glucose tolerance. The hypoglycemic effect of BBOV is similar to that of bis (maltolato) oxovanadium but is less toxic in median lethal dose. Overall, the present work will provide useful information for further development of new anti-diabetic vanadium compounds.


A systematic review of magnetic versus conventional ureteric stents for short term ureteric stenting.
Ureteric stents play an essential role in urology. However, patients can suffer a range of stent-related symptoms with stent in situ and during removal. Conventional ureteric stents are removed using a flexible cystoscopy, whereas magnetic stents may be rapidly removed with a smaller catheter-like retrieval device. The primary aim of this systematic review was to compare the morbidity including pain associated with conventional versus magnetic ureteric stents. The secondary aim was cost comparison. Searches were performed across databases, including Medline, Scopus, Embase and Cochrane. This review was performed in accordance with Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA). The search from the 5 databases returned a total of 358 articles. After duplicates were removed as well as the inclusion and exclusion criteria applied, a total of 6 studies were included in the final review. Ureteric Stent Symptoms Questionnaire (USSQ) and Visual Analogue Score (VAS) were used in most of the studies. All the studies reported that magnetic ureteric stents resulted in a reduction in the pain on the removal of magnetic ureteric stents, and no statistically significant difference with indwelling ureteric stents. Furthermore, majority of the studies reported a reduction in the cost associated with magnetic ureteric stents. There is no significant difference in pain from indwelling ureteric stents. There is a reduction in pain with the removal of magnetic ureteric stents compared to conventional removal via cystoscopy and an associated reduction in cost.


Direct measurement of electrical transport through single DNA molecules of complex sequence.
Seemingly contradicting results raised a debate over the ability of DNA to transport charge and the nature of the conduction mechanisms through it. We developed an experimental approach for measuring current through DNA molecules, chemically connected on both ends to a metal substrate and to a gold nanoparticle, by using a conductive atomic force microscope. Many samples could be made because of the experimental approach adopted here, which enabled us to obtain reproducible results with various samples, conditions, and measurement methods. We present multi-leveled evidence for charge transport through 26-bp-long dsDNA of a complex sequence, characterized by S-shaped current-voltage curves that show currents >220 nA at 2 V. This significant observation implies that a coherent or band transport mechanism takes over for bias potentials leading to high currents (>1 nA).


CircLIFR synergizes with MSH2 to attenuate chemoresistance via MutSα/ATM-p73 axis in bladder cancer.
Cisplatin (CDDP) has become a standard-of-care treatment for muscle-invasive bladder cancer (MIBC), while chemoresistance remains a major challenge. Accumulating evidence indicates that circular RNAs (circRNAs) are discrete functional entities. However, the regulatory functions as well as complexities of circRNAs in modulating CDDP-based chemotherapy in bladder cancer are yet to be well revealed. Through analyzing the expression profile of circRNAs in bladder cancer tissues, RNA FISH, circRNA pull-down assay, mass spectrometry analysis and RIP, circLIFR was identified and its interaction with MSH2 was confirmed. The effects of circLIFR and MSH2 on CDDP-based chemotherapy were explored by flow cytometry and rescue experiments. Co-IP and Western blot were used to investigate the molecular mechanisms underlying the functions of circLIFR and MSH2. Biological implications of circLIFR and MSH2 in bladder cancer were implemented in tumor xenograft models and PDX models. CircLIFR was downregulated in bladder cancer and expression was positively correlated with favorable prognosis. Moreover, circLIFR synergizing with MSH2, which was a mediator of CDDP sensitivity in bladder cancer cells, positively modulated sensitivity to CDDP in vitro and in vivo. Mechanistically, circLIFR augmented the interaction between MutSα and ATM, ultimately contributing to stabilize p73, which triggered to apoptosis. Importantly, MIBC with high expression of circLIFR and MSH2 was more sensitive to CDDP-based chemotherapy in tumor xenograft models and PDX models. CircLIFR could interact with MSH2 to positively modulate CDDP-sensitivity through MutSα/ATM-p73 axis in bladder cancer. CircLIFR and MSH2 might be act as promising therapeutic targets for CDDP-resistant bladder cancer.


Rad23 and Rpn10: perennial wallflowers join the melee.
Substrate ubiquitination is highly regulated; by contrast, substrate targeting to the proteasome has been considered a stochastic process that is mediated primarily by high-affinity interaction with multi-ubiquitin chains. However, recent findings have shown that substrate recognition by the proteasome is also regulated, and requires Rad23, Dsk2 and Rpn10. These studies suggest that the engagement of protein ubiquitination and translocation with degradation by the proteasome is coordinated by a series of regulated events.


TDP-43 Puts the STING in ALS.
In a recent study, Yu et al. demonstrated that TAR DNA-binding protein of 43 kDa (TDP-43) causes inflammation in amyotrophic lateral sclerosis (ALS) by triggering mitochondrial (mt)DNA release into the cytoplasm, which subsequently activates the cytoplasmic DNA-sensing cyclic GMP-AMP synthase (cGAS)/stimulator of interferon genes (STING) pathway. These results suggest that inhibition of cGAS/STING could help mitigate inflammation-related neuropathology in ALS.