Characterization of Phytoestrogens in Medicago sativa L. and Grazing Beef Cattle.
Phytoestrogens are plant-produced bioactive secondary metabolites known to play an integral role in plant defense that frequently accumulate in times of stress and/or microbial infection. Phytoestrogens typically belong to two distinct chemical classes; flavonoids (isoflavones) and non-flavonoids (lignans and coumestans). Upon consumption by livestock, high concentrations of phytoestrogens can cause long-term disruption in reproduction due to structural similarities with mammalian estrogens and their tendency to bind estrogen receptors. Wide variation in phytoestrogen concentration has been reported in pasture legumes and corresponding silage or hay. Lucerne is a common perennial pasture legume in temperate climates, but information on phytoestrogen production or accumulation in grazing livestock is currently limited. Therefore, metabolic profiling using UHPLC-MS-QToF was performed to identify and quantitate key phytoestrogens in both fresh and dried lucerne fodder from replicated field or controlled glasshouse environments. Phytoestrogens were also profiled in the blood plasma of Angus cattle grazing field-grown lucerne. Results revealed that phytoestrogens varied quantitatively and qualitatively among selected lucerne cultivars grown under glasshouse conditions. Fresh lucerne samples contained higher concentrations of coumestans and other phytoestrogenic isoflavones than did dried samples for all cultivars profiled, with several exceeding desirable threshold levels for grazing cattle. Coumestans and isoflavones profiled in plasma of Angus heifers grazing lucerne increased significantly over a 21-day sampling period following experimental initiation. Currently, threshold concentrations for phytoestrogens in plasma are unreported. However, total phytoestrogen concentration exceeded 300 mg·kg-1 in fresh and 180 mg·kg-1 in dried samples of selected cultivars, suggesting that certain genotypes may upregulate phytoestrogen production, while others may prove suitable sources of fodder for grazing livestock.


Characteristic mutations found in the ML0411 gene of Mycobacterium leprae isolated in Northeast Asian countries.
Genome analysis of Mycobacterium leprae strain Kyoto-2 in this study revealed characteristic nucleotide substitutions in gene ML0411, compared to the reference genome M. leprae strain TN. The ML0411 gene of Kyoto-2 had six SNPs compared to that of TN. All SNPs in ML0411 were non-synonymous mutations that result in amino acid replacements. In addition, a seventh SNP was found 41 bp upstream of the start codon in the regulatory region. The seven SNP sites in the ML0411 region were investigated by sequencing in 36 M. leprae isolates from the Leprosy Research Center in Japan. The SNP pattern in 14 of the 36 isolates showed similarity to that of Kyoto-2. Determination of the standard SNP types within the 36 stocked isolates revealed that almost all of the Japanese strains belonged to SNP type III, with nucleotide substitutions at position 14676, 164275, and 2935685 of the M. leprae TN genome. The geographical distribution pattern of east Asian M. leprae isolates by discrimination of ML0411 SNPs was investigated and interestingly turned out to be similar to that of tandem repeat numbers of GACATC in the rpoT gene (3 copies or 4 copies), which has been established as a tool for M. leprae genotyping. All seven Korean M. leprae isolates examined in this study, as well as those derived from Honshu Island of Japan, showed 4 copies of the 6-base tandem repeat plus the ML0411 SNPs observed in M. leprae Kyoto-2. They are termed Northeast Asian (NA) strain of M. leprae. On the other hand, many of isolates derived from the Okinawa Islands of Japan and from the Philippines showed 3 copies of the 6-base tandem repeat in addition to the M. leprae TN ML0411 type of SNPs. These results demonstrate the existence of M. leprae strains in Northeast Asian region having characteristic SNP patterns.


Prepregnancy lipids related to preterm birth risk: the coronary artery risk development in young adults study.
Preterm birth is associated with maternal cardiovascular risk, but mechanisms are unknown. We considered that dyslipidemia may predispose women to both conditions and that prepregnancy lipids may be related to preterm birth risk. We hypothesized that low or high prepregnancy plasma lipids would be associated with preterm birth. A total of 1010 women (49% black) enrolled in the multicenter, prospective Coronary Artery Risk Development in Young Adults study with at least one singleton birth during 20 yr of follow-up were evaluated. Postbaseline preterm births less than 34 wk or 34 to less than 37 wk vs. greater than 37 wk gestation. We detected a U-shaped relationship between prepregnancy cholesterol concentrations and preterm birth risk. Women with prepregnancy cholesterol in the lowest quartile compared with the second quartile (<156 vs. 156-171 mg/dl) had an increased risk for preterm birth 34 to less than 37 wk (odds ratio 1.86; 95% confidence interval 1.10, 3.15) and less than 34 wk (odds ratio 3.04; 1.35, 6.81) independent of race, age, parity, body mass index, hypertension during pregnancy, physical activity, and years from measurement to birth. Prepregnancy cholesterol in the highest quartile (>195 mg/dl) was also associated with preterm birth less than 34 wk among women with normotensive pregnancies (odds ratio 3.80; 95% confidence interval 1.07, 7.57). There were no associations between prepregnancy triglycerides, low-density lipoprotein cholesterol, or high-density lipoprotein cholesterol and preterm birth. Both low and high prepregnancy cholesterol were related to preterm birth risk. These may represent distinct pathways to the heterogeneous outcome of preterm birth. Additional studies are needed to elucidate mechanisms that link low or high cholesterol to preterm birth and later-life sequelae.


End or means--the "what" and "how" of observed intentional actions.
Action understanding and learning are suggested to be mediated, at least in part, by the human mirror neuron system (hMNS). Static images as well as videos of actions with the outcome occluded have been shown to activate the hMNS. However, whether the hMNS preferentially responds to end or means of an action remains to be investigated. We, therefore, presented subjects with videos of intentional actions that were shown from two perspectives (factor 1, perspective: first vs. third person) while subjects directed their attention to the means or the end thereof (factor 2, task: means vs. end). End- or means-related changes in BOLD signal and corticospinal excitability (CSE) were assessed using fMRI and TMS, respectively. Judging the means of an action compared with its end differentially activated bilateral ventral premotor (vPMC) and inferior parietal cortex (IPL), that is, the core regions of the hMNS. The reverse contrast revealed left precuneus and bilateral superior frontal, angular, and middle temporal gyrus activity. In accordance, the two tasks, although identically in stimulus properties, modulated CSE differentially. Although recent studies suggest that the hMNS may prefer the presence of a goal or context, our data show that within the same context, it responds preferentially when attention is directed to the action means. Consequently, in addition to inferring action goals, a key function of the hMNS may be to anticipate the trajectories and dynamics of observed actions, which is a prerequisite for any timely interaction.


Compensation of blue phase I by blue phase II in optoeletronic device.
Compensation effect of blue phase I (BP I) with blue phase II (BP II) liquid crystal was demonstrated. BP I and BP II were co-exist in the optoeletronic device by polymer stabilization. Consequently, disadvantages of BP I and BP II were greatly improved by compensation effect and resulted in high contrast ratio, low hysteresis and fast falling time. Mechanism of compensation effect was explained by relaxation ability of lattice structure under electrical field and compensation structure was well confirmed by Bragg's reflectance spectrum and Commission International de l'Éclairage chromaticity diagram.


Ectogalactosyltransferase. Presence of enzyme and acceptors on the rat lymphocyte cell surface.
Experiments are described to demonstrate the existence of ectogalactosyltransferase activity on the lymphocyte surface. The procedures described enable us to exclude the possibility of misleading results due to precursor hydrolysis and intracellular utilization of the free galactose. This depicted transferase is able to catalyse the transfer of a galactosyl residue from UDP-galactose to a nonphagocytosable exogenous acceptor and to endogenous membrane acceptors. The cells galactosylated in this way acquired new agglutinating properties with soybean agglutinin, which proves the external position of the galactosyl residues incorporated on the cell surface.


Nanoscale π-conjugated ladders.
It is challenging to increase the rigidity of a macromolecule while maintaining solubility. Established strategies rely on templating by dendrons, or by encapsulation in macrocycles, and exploit supramolecular arrangements with limited robustness. Covalently bonded structures have entailed intramolecular coupling of units to resemble the structure of an alternating tread ladder with rungs composed of a covalent bond. We introduce a versatile concept of rigidification in which two rigid-rod polymer chains are repeatedly covalently associated along their contour by stiff molecular connectors. This approach yields almost perfect ladder structures with two well-defined π-conjugated rails and discretely spaced nanoscale rungs, easily visualized by scanning tunnelling microscopy. The enhancement of molecular rigidity is confirmed by the fluorescence depolarization dynamics and complemented by molecular-dynamics simulations. The covalent templating of the rods leads to self-rigidification that gives rise to intramolecular electronic coupling, enhancing excitonic coherence. The molecules are characterized by unprecedented excitonic mobility, giving rise to excitonic interactions on length scales exceeding 100 nm. Such interactions lead to deterministic single-photon emission from these giant rigid macromolecules, with potential implications for energy conversion in optoelectronic devices.


Improvement in preoperative staging of gastric adenocarcinoma with positron emission tomography.
Positron emission tomography (PET) with 18- fluorodeoxyglucose (FDG) has been used to both detect and stage a variety of malignancies. The current study examined the value of PET for preoperative staging of gastric adenocarcinoma. Sixty-eight patients (49 males and 19 females) with gastric adenocarcinoma, who were referred for preoperative FDG-PET scans, were enrolled in this study. The patients underwent spiral-computed tomography (CT) within 1 week of referral. The final diagnosis in all patients was made by histologic and surgical findings. For quantitative PET analysis, the regional tumor FDG uptake was measured by the standardized uptake value (SUV). For the primary tumor of a gastric adenocarcinoma, PET demonstrated an increased uptake in 64 of 68 patients (sensitivity, 94%), with a mean SUV of 7.0 (range, 0.9-27.7). A comparison of FDG uptake and clinicopathologic features showed significant association between FDG uptake and macroscopic type, tumor size, lymph node metastasis, histologic type, and TNM stage. The PET scan had a similar accuracy with that of CT for diagnosing local and distant lymph node metastases as well as peritoneal status. In assessing local lymph node status, however, PET had a higher specificity than CT (92% vs. 62%, P = 0.000). Moreover, PET had additional diagnostic value in 10 (15%) of 68 patients by upstaging 4 (6%) and downstaging 6 (9%) patients. PET combined with CT was more accurate for preoperative staging than either modality alone (66% vs. 51%, 66% vs. 47%, respectively; P = 0.002). FDG-PET improves the preoperative TNM staging of gastric adenocarcinoma. Based on its superior specificity, FDG-PET can facilitate the selection of patients for a curative resection by confirming a nodal status identified by CT.


[Reversible hypertransaminasemia after diet therapy].
An increase in transaminases may be due to multiple causes. We present a case of reversible hypertransaminasemia following specific dietary therapy. The various potential aetiopathogenies are reviewed to explain the raising of transaminase levels and the improvement in analytical results after the institution of this dietary treatment.


[Fish nutrition. Brief review for the practitioner regarding food and ornamental fish].
In behalf of the practitioner a short survey is given about nutrition of fish and shrimps. Interrelations between fish development and influences by their water ecosystem concerning temperature, mineral and gas levels, waterflow et cetera together with the supply of food are mentioned. Further comments are given about common nutritional needs of different species in connection with growth, bodyweight et cetera and purpose of fishkeeping for instance for consumption, reproduction or ornamentally in different systems from aquarium to lagoon. More specifically feed formulation from raw materials to complete diets and nutritional analyses in respect to allowances for maintenance and growth are discussed. Data about fresh- and saltwater species from tropical to arctic circumstances mentioning trout, salmon, catfish, carp, sea bass, shrimp et cetera are given. Some remarks about manufacturing, and application of complete compound feeds are added to the survey. In behalf of further study or for enhancement of knowledge a number of books are recommended in a list of references.


TSLP is involved in expansion of early thymocyte progenitors.
Thymic stromal derived lymphopoietin (TSLP) is preferentially and highly expressed in the thymus, but its function in T cell development is not clear. We report here that TSLP, independently or in combination with IL-7, enhances thymopoiesis in the murine fetal thymic organ culture (FTOC) model. Furthermore, TSLP preferentially increases the number and proliferation of the (DN1 and DN2) pro-T progenitor cells, and FTOC lobes from TSLP receptor-null mice show a decreased number of these cells. Finally, DN1-DN2 cells expanded with TSLP in vitro are functional T progenitors that are able to differentiate into mature T cells in fetal or adult thymus organs. Together, these data suggest that TSLP plays an important role in expansion of thymocyte progenitors and may be of value for expanding T progenitor cells in vitro.


Considerations for personalized surgery in patients with papillary thyroid cancer.
Personalizing treatment for papillary thyroid cancer (PTC) requires a multidisciplinary approach. The surgical management of PTC has long been based on retrospective studies focusing on endpoints that are of debatable significance. There is considerable debate in the literature regarding the optimal initial treatment for PTC. Many of these issues are discussed in this review. These debates have hindered the development of a tailored treatment strategy. The ability to optimally personalize a surgical plan for the treatment of PTC is ultimately dependent on an understanding of the biological behavior of that individual patient's tumor. We are at the genesis of an age where molecular biology advances endeavor to profile a patient's tumor behavior. This review summarizes current strategies for managing PTC, where we are with personalizing surgery for these patients, and where we hope to go. Thyroid surgery is one of the newest fields for the application of minimally invasive techniques and can now be accomplished endoscopically or with robotic assistance in many patients who therefore may benefit from these alternative approaches. When treating a patient with PTC, it seems prudent to weigh the many factors discussed in this review to individualize the most optimal surgical plan.


Phytochemicals protect L02 cells against hepatotoxicity induced by emodin via the Nrf2 signaling pathway.
Dihydromyricetin (DMY), hyperoside and silybin are phytochemicals that belong to a class called flavonoids, and they have been used in liver protection pharmaceutical preparations, but the specific mechanism of these chemicals is still unclarified. This study aims to investigate the hepatoprotective effects and potential mechanism of these phytochemicals. The immortalized human hepatocyte cell line L02 was treated with 200 μM emodin for 48 h, and this was used as a hepatocyte injury model. The L02 cells were treated with both 200 μM emodin and different concentrations of DMY/hyperoside/silybin for 48 h to investigate the protective effects of these phytochemicals. The CCK-8 assay was used to detect cell viability. RT-qPCR and western blotting were performed to examine the mRNA and protein expression, respectively, of the classic bile acid synthetic pathway gene CYP7A1, the bile acid efflux transporter bile salt export pump (BSEP), the nuclear factor erythroid-2-related factor 2 (Nrf2) and the drug processing gene CYP1A2. DMY, hyperoside and silybin prevented the impairment of cell viability that was caused by emodin-induced hepatotoxicity in a dose-dependent manner, and at a low concentration (10 μM), the protective effect followed the order hyperoside > DMY > silybin, while at a high concentration (160 μM), the protective effect followed the order DMY > hyperoside > silybin. These phytochemicals reduced the expression of CYP7A1 at both the mRNA and protein levels. BSEP was not influenced by the phytochemical intervention. When 200 μM emodin was used for 48 h with the addition of the phytochemicals at 200 μM, the nuclear protein expression of Nrf2 significantly increased and CYP1A2 expression decreased. DMY, hyperoside and silybin prevented the hepatotoxicity induced by emodin in the L02 cells, potentially, via the Nrf2 signaling pathway.


Enteric Pathogens Exploit the Microbiota-generated Nutritional Environment of the Gut.
Host bacterial associations have a profound impact on health and disease. The human gastrointestinal (GI) tract is inhabited by trillions of commensal bacteria that aid in the digestion of food and vitamin production and play crucial roles in human physiology. Disruption of these relationships and the structure of the bacterial communities that inhabit the gut can contribute to dysbiosis, leading to disease. This fundamental relationship between the host and microbiota relies on chemical signaling and nutrient availability and exchange. GI pathogens compete with the endogenous microbiota for a colonization niche (1, 2). The ability to monitor nutrients and combine this information with the host physiological state is important for the pathogen to precisely program the expression of its virulence repertoire. A major nutrient source is carbon, and although the impact of carbon nutrition on the colonization of the gut by the microbiota has been extensively studied, the extent to which carbon sources affect the regulation of virulence factors by invading pathogens has not been fully defined. The GI pathogen enterohemorrhagic E. coli (EHEC) gages sugar sources as an important cue to regulate expression of its virulence genes. EHEC senses whether it is in a gluconeogenic versus a glycolytic environment, as well as fluctuations of fucose levels to fine tune regulation of its virulence repertoire.


Long-term Exercise Adherence After High-intensity Interval Training in Cardiac Rehabilitation: A Randomized Study.
Exercise adherence in general is reported to be problematic after cardiac rehabilitation. Additionally, vigorous exercise is associated with impaired exercise adherence. As high-intensity interval training (HIT) is frequently used as a therapy to patients with coronary artery disease in cardiac rehabilitation, the objective was to assess long-term exercise adherence following an HIT cardiac rehabilitation programme. A multicentre randomized study was carried out. Eligible participants were adults who had previously attended a 12-week HIT cardiac rehabilitation programme, as either a home-based or hospital-based HIT (treadmill exercise or group exercise). The primary outcome was change in peak oxygen uptake; secondary outcomes were self-reported and objectively measured physical activity. Out of 83 eligible participants, 76 were available for assessment (68 men/8 women, mean age 59 (8) years) at a one-year follow-up. Peak oxygen uptake was significantly elevated above baseline values, (treadmill exercise: 35.8 (6.4) vs. 37.4 (7.4) ml kg(-1) min(-1) , group exercise: 32.7 (6.5) vs. 34.1 (5.8) ml kg(-1) min(-1) and home-based exercise: 34.5 (4.9) vs. 36.7 (5.8) ml kg(-1) min(-1) at baseline and follow-up, respectively), with no significant differences between groups. The majority of the participants (>90%) met the recommended daily level of 30 minutes of moderate physical activity. The home-based group showed a strong trend towards increased physical activity compared with the hospital-based groups. The results from this study have shown that both home-based and hospital-based HIT in cardiac rehabilitation induce promising long-term exercise adherence, with maintenance of peak oxygen uptake significantly above baseline values at a one-year follow-up. The implication for physiotherapy practice is that HIT in cardiac rehabilitation induces satisfactory long-term exercise adherence.


Real-time monitoring of apoptosis by caspase-3-like protease induced FRET reduction triggered by amyloid aggregation.
Amyloid formation is cytotoxic and can activate the caspase cascade. Here, we monitor caspase-3-like activity as reduction of fluorescence resonance energy transfer (FRET) using the contstruct pFRET2-DEVD containing enhanced cyan fluorescent protin (EYFP) linked by the caspase-3 specific cleavage site residues DEVD. Beta-TC-6 cells were transfected, and the fluoorescence was measured at 440 nm excitation and 535 nm (EYFP) and 480 nm (ECFP) emission wavelength. Cells were incubated with recombinant pro lset Amyloid Polypeptide (rec prolAPP) or the processing metabolites of prolAPP; the N-terminal flanking peptide withIAPP (recN+IAPP); IAPP with the C-terminal flanking peptied (recIAPP+C) and lslet Amyloid Polypeptide (recIAPP) . Peptides were added in solubilized from (50 microM) or as performed amyloid-like fibrils, or as a combination of these. FRET was measured and incubation with a mixture of solubilized peptide and performed fibrils resulted in loss of FRET and apoptosis was determined to occure in cells incubated with recproIAPP (49%), recN+IAPP (46%), recIAPP (72%) and recIAPP+C (59%). These results show that proIAPP and the processing intermediates reside the same cell toxic capacity as IAPP, and they can all have a central role in the reduction of beta-cell number in type 2 diabetes.


Incidental traumatic pseudoaneurysm of the middle meningeal artery: case report and literature review.
Traumatic pseudoaneurysm of the middle meningeal artery is rare and is associated with high mortality. Skull fracture is usually an associated feature of this entity. An elderly male was brought to our hospital in a stage of coma. The details of ictus were not known. The Glasgow coma scale score was 9/15. Examination revealed a pseudo-aneurysm arising from the posterior branch of the left middle meningeal artery which was excised. The case is presented for its rarity and its characteristic radiology. Traumatic pseudoaneurysm of middle meningeal artery is rare. It is important to recognize this treatable entity.


Research actuality in the genetics of stroke.
Stroke can be viewed as a paradigm for late-onset, complex polygenic diseases. There are two main clinical phenotypes for stroke: ischemic stroke, responsible for 80-90% of stroke events, and hemorrhagic stroke, responsible for the remaining 10-20%. Stroke may either be the outcome of a number of monogenic disorders or, more commonly, a polygenic multifactorial disease. Cerebral autosomal dominant arteriopathy with subcortical infarcts and leucoencephalopathy (CADASIL), due to mutations in the Notch 3 gene, is the best example of monogenic pathology leading to stroke. The identification of individual causative mutations for polygenic stroke is problematic due to the complexity of such condition. The two main methods of genetic investigation are linkage analyses and association studies, each with advantages and limitations. Associations with polymorphisms in a variety of candidate genes have been investigated, including hemostatic genes, genes controlling homocystein metabolism, the angiotensin-converting enzyme gene, and the endothelial nitric oxide synthase gene. The combination of linkage and association approaches has led to the identification of the first putative gene associated with common polygenic stroke, PDE4D, mapped to chromosome 5q21. The biological revolution of the past years, spurred by the Human Genome Project, promises the advent of novel technologies supported by bioinformatics, which will transform the study of polygenic disorders such as stroke. Understanding the causes of stroke and its effect will allow definition of high-risk populations and make possible specific programs of primary and secondary prevention as well as new therapeutic approaches where prevention has failed.


Understanding the Mysterious M2 Macrophage through Activation Markers and Effector Mechanisms.
The alternatively activated or M2 macrophages are immune cells with high phenotypic heterogeneity and are governing functions at the interface of immunity, tissue homeostasis, metabolism, and endocrine signaling. Today the M2 macrophages are identified based on the expression pattern of a set of M2 markers. These markers are transmembrane glycoproteins, scavenger receptors, enzymes, growth factors, hormones, cytokines, and cytokine receptors with diverse and often yet unexplored functions. This review discusses whether these M2 markers can be reliably used to identify M2 macrophages and define their functional subdivisions. Also, it provides an update on the novel signals of the tissue environment and the neuroendocrine system which shape the M2 activation. The possible evolutionary roots of the M2 macrophage functions are also discussed.


How HIV diagnosis and disclosure affect sexual behavior and relationships in Ugandan fishing communities.
In this article we examine how members of fishing communities on the shores of Lake Victoria in Uganda respond to HIV diagnosis in terms of disclosure to sexual partners. We then explore the subsequent changes in sexual behavior and relationships. To access this information, we collected life history data from 78 HIV-positive individuals in five fishing communities. We found that the strength of the sexual relationships shaped how and why individuals disclosed to partners, and that these relationships tended to be stronger when partners shared familial responsibility. Those who perceived their current sexual partnership to be weak sought to conceal their status by maintaining prediagnosis patterns of sexual behavior. The majority of the study's participants rarely changed their sexual behavior following HIV diagnosis, regardless of their relationship's strength. These findings elucidate barriers to disclosure and behavior change, and suggest that a life-course approach might enhance individual-level counseling so that counselors can provide tailored support to individuals regarding disclosure decisions and outcomes.


Using collective intelligence to fine-tune public health policy.
The European Union Future Internet Assembly, the roadmap for the Web heading towards semantic interoperability and building on the UK's adoption of the Internet and social media are accelerating the development of Web 3.0. A number of health portals are opening, some with facilities for the capture of Patient Based Records. Collective Intelligence will be generated that, applied to health, has potential to support Public Health policy. By using the Internet, millions of people in the course of their daily activities contribute to uncertified data stores, some explicitly collaborating to create collective knowledge bases, some contributing implicitly through the patterns of their choices and actions. An application of soft computing, called Collective Health Intelligence, that reasons uncertified and certified data could enhance the social pool of existing health knowledge available to the public health agencies. Collective Health Intelligence could be used to complement national programmes by employing innovative sampling techniques, cost-effectively generating anonymous data trends that would quantify policy, indicate epidemiological effects and supply metrics to test policy efficacy.


The interaction between strigolactones and other plant hormones in the regulation of plant development.
Plant hormones are small molecules derived from various metabolic pathways and are important regulators of plant development. The most recently discovered phytohormone class comprises the carotenoid-derived strigolactones (SLs). For a long time these compounds were only known to be secreted into the rhizosphere where they act as signaling compounds, but now we know they are also active as endogenous plant hormones and they have been in the spotlight ever since. The initial discovery that SLs are involved in the inhibition of axillary bud outgrowth, initiated a multitude of other studies showing that SLs also play a role in defining root architecture, secondary growth, hypocotyl elongation, and seed germination, mostly in interaction with other hormones. Their coordinated action enables the plant to respond in an appropriate manner to environmental factors such as temperature, shading, day length, and nutrient availability. Here, we will review the current knowledge on the crosstalk between SLs and other plant hormones-such as auxin, cytokinin, abscisic acid (ABA), ethylene (ET), and gibberellins (GA)-during different physiological processes. We will furthermore take a bird's eye view of how this hormonal crosstalk enables plants to respond to their ever changing environments.


Physiological properties of central medial and central lateral amygdala neurons.
Mounting evidence implicates the central (CE) nucleus of the amygdala in the mediation of classically conditioned fear responses. However, little data are available regarding the intrinsic membrane properties of CE amygdala neurons. Here, we characterized the physiological properties of CE medial (CE(M)) and CE lateral (CE(L)) amygdala neurons using whole cell recordings in brain slices maintained in vitro. Several classes of CE neurons were distinguished on the basis of their physiological properties. Most CE(M) cells (95%), here termed "late-firing neurons," displayed a marked voltage- and time-dependent outward rectification in the depolarizing direction. This phenomenon was associated with a conspicuous delay between the onset of depolarizing current pulses and the first action potential. During this delay, the membrane potential (V(m)) depolarized slowly, the steepness of this depolarizing ramp increasing as the prepulse V(m) was hyperpolarized from -60 to -90 mV. Low extracellular concentrations of 4-aminopyridine (30 microM) reversibly abolished the outward rectification and the delay to firing. Late-firing CE(M) neurons displayed a continuum of repetitive firing properties with cells generating single spikes at one pole and high-frequency (> or =90 Hz) spike bursts at the other. In contrast, only 56% of CE(L) cells displayed the late-firing behavior prevalent among CE(M) neurons. Moreover, these CE(L) neurons only generated single spikes in response to membrane depolarization. A second major class of CE(L) cells (38%) lacked the characteristic delay to firing observed in CE(M) cells, generated single spikes in response to membrane depolarization, and displayed various degrees of inward rectification in the hyperpolarizing direction. In both regions of the CE nucleus, two additional cell types were encountered infrequently (< or =6% of our samples). One type of neurons, termed "low-threshold bursting cells" had a behavior reminiscent of thalamocortical neurons. The second type of cells, called "fast-spiking cells," generated brief action potentials at high rates with little spike frequency adaptation in response to depolarizing current pulses. These findings indicate that the CE nucleus contains several types of neurons endowed with distinct physiological properties. Moreover, these various cell types are not distributed uniformly in the medial and lateral sector of the CE nucleus. This heterogeneity parallels anatomic data indicating that these subnuclei are part of different circuits.


Aesthetic rehabilitation of oligodontia in primary dentition with adhesive partial denture.
The primary teeth are essential for bone development and establishment of the arches on occlusion. Thus, the congenitally absence of teeth may trigger a shift in the balance of the occlusion, promoting disharmony in the structures of the maxilla-mandibular system. However, some interventions are possible to be performed in these cases even in pediatric patients, to redirect growth, preventing growth deviations and reestablishing the aesthetic. The aim of this paper is to report the treatment of a 4-year-old child presenting congenitally absence of mandibular central and lateral incisors and maxilla lateral incisors, which consequently compromises aesthetics, occlusal function, and the development and the functional growth of the bones. The oral rehabilitation was performed with an adhesive partial denture, which was able to restore the aesthetic and the occlusal function, therefore being a viable alternative in the treatment of this patient of little age.


PaLS Study: Tobacco, Alcohol and Drugs Usage among Polish University Students in the Context of Stress Caused by the COVID-19 Pandemic.
The COVID-19 pandemic and imposed restrictions were strong stress factors for young people, especially students. Increased alcohol consumption, smoking cigarettes, usage of heated tobacco products, and other stimulants are common methods of coping with anxiety. However, they can have serious negative health effects. A survey consisting of 12 questions related to mental health and psychoactive substance taking habits was distributed among Polish students between 22 February 2021 and 3 April 2021. A total of 1323 participants met all inclusion criteria (nfemales = 1021, nmales = 297, nother gender = 5). The mean age was 22 years old (±4.17); 47.62% were medical university students. A total of 71.92% reported negative impact, 8.25% did not notice changes, and 12.58% declared a positive pandemic impact on their mental health. A total of 12.58% declared an increase, 70.22% did not see any differences, and 17.20% reported a decrease in their psychoactive substance usage tendency due to the pandemic. Worse perceived psychologic well-being was correlated with a higher tendency to use tobacco (p < 0.001) and alcohol (p < 0.001), and not with marijuana and products containing tetrahydrocannabinol (p = 0.136), and hard drugs (p = 0.799). The majority of participants declared a negative pandemic impact on mental health and did not report significant changes in psychoactive substance taking habits. Medical personnel should be aware of the current situation and apply for proper prevention and treatment programs.


Dose-response study of ultraviolet radiation for induction of photoallergy to tetrachlorosalicylanilide in guinea pigs.
The dose of ultraviolet light (UV) required to induce photoallergy to tetrachlorosalicylanilide (T4CS) in guinea pigs was studied. A simple induction technique consisting of only one treatment with adjuvant, chemical and UV was used. The concentration of T4CS, which was administered either topically or intradermally, was low to reduce self-screening of UV by the chemical. Doses of UVA from fluorescent blacklamps ranged from 0.4 to 300 kJ/m2. All animals were challenged with topically applied T4CS and 40 kJ/m2 UV. The results showed that incidence of photoallergy increased with increasing dose of UV and reached a plateau between 4.5-45 kJ/m2 following intradermal T4CS and 4-120 kJ/m2 following topical T4CS. The studies demonstrated for the first time decreased response following larger induction doses of UV; photoallergy was not induced by the largest induction dose of 300 kJ/m2. Attenuation of UVB during induction did not affect the decrease. The lack of response may be attributable to further photochemical reactions of a T4CS-protein conjugate with loss of antigenic properties, or to immune suppression initiated by either UVA or an interaction of T4CS and UVA.


Forced treadmill running suppresses postincisional pain and inhibits upregulation of substance P and cytokines in rat dorsal root ganglion.
Exercise causes a variety of psychophysical effects (eg, alterations in pain sensation). Tissue injury induces mediator releases in the spinal cord resulting in pain hypersensitivity; however, the contribution of the dorsal root ganglion (DRG) is poorly understood. In this study, we tested if forced treadmill running can attenuate postoperative pain and alter substance P (SP) or proinflammatory cytokine level in the DRG by using a rat model of skin/muscle incision and retraction (SMIR). We evaluated mechanical sensitivity to von Frey stimuli (6 and 15 g) and expression of SP, interleukin-1β, and interleukin-6 in the DRG of sham-operated sedentary rats, SMIR sedentary rats, sham-operated rats with forced treadmill running, and SMIR rats with forced treadmill running. At postoperative day 8, trained rats ran for 5 days per week for 4 weeks on a treadmill 70 minutes/d with an intensity of 18 m/min. On postoperative day 6, SMIR sedentary rats displayed a significant mechanical hypersensitivity that persisted until postoperative day 35. By comparison, SMIR-operated rats, which received forced treadmill running, exhibited a quick recovery from mechanical hypersensitivity. SMIR sedentary rats showed an upregulation of SP, interleukin-1β, and interleukin-6 in the DRG at postoperative days 14 and 28, whereas SMIR-operated rats receiving forced treadmill running reversed this upregulation at postoperative day 28. We concluded that forced treadmill running alleviated persistent postincisional pain caused by SMIR surgery. This appears to be protective against postoperative pain, which probably relates to the downturn in excess SP, interleukin-1β, and interleukin-6 in the DRG. Controlling the expression of SP, interleukin-6, and interleukin-1β in the DRG can help manage postoperative pain. This finding could potentially help clinicians and physical therapists who seek to examine how exercise may attenuate postsurgical pain and its mechanism.


Interleukin-10 genotypes are associated with systemic sclerosis and influence disease-associated autoimmune responses.
Systemic sclerosis (SSc; scleroderma) is a connective tissue disease, characterized by fibrotic, immunological, and vascular abnormalities. Interleukin-10 (IL-10) is an anti-inflammatory cytokine that modulates collagen production and B-cell survival. To determine if certain IL-10 genotypes are risk factors for the development of SSc and influence disease-associated autoimmune responses, 248 Caucasian and 264 Japanese SSc patients and controls were genotyped for three loci: -3575, -2849, and -2763. Sera from patients were characterized for SSc-associated autoantibodies. In Caucasians, at -3575 and -2763, the frequency of AA homozygotes was higher in patients as compared with controls (P=0.0005; P=0.002). In Japanese subjects, the frequency of AC heterozygotes at -2763 was higher, and that of CC homozygotes lower, in patients with diffuse SSc as compared to controls (P=0.04). Particular IL-10 genotypes were associated with SSc-related autoantibodies. These results suggest that IL-10 genotypes contribute to the etiology of scleroderma.


Structure prediction of physiological bis(amino acidato)copper(II) species in aqueous solution: The copper(II) compounds with l-glutamine and l-histidine.
Neutral (l-histidinato)(l-glutaminato)copper(II) [Cu(His)(Gln)] has been established as the most abundant ternary copper(II) amino acid compound of the exchangeable copper(II) pool in blood plasma. The experimental studies of Cu(His)(Gln) and bis(glutaminato)copper(II) [Cu(Gln)2] in solutions did not specify their complete geometries. To determine the geometries, this paper investigates the conformers, energy landscapes, and a structure-magnetic parameters relation of Cu(Gln)2 and Cu(His)(Gln) by the density functional theory (DFT) calculations. We assume a glycine-like coordination of Gln (other coordination patterns are dismissed because of steric reasons), and three His in-plane copper(II) binding modes. The conformational analyses are performed in the gas phase and implicitly modeled aqueous solution. The reliability of the DFT relative electronic and Gibbs free energies of the Cu(His)(Gln) conformers is confirmed by benchmarking against the corresponding energies obtained by the domain-based local pair natural orbital coupled-cluster method with singles, doubles, and perturbative triples [DLPNO-CCSD(T)]. Several cis- and trans-Cu(His)(Gln) conformers with His in the histaminate-like and glycine-like modes have low Gibbs free energies, and the greatest estimated metal-binding affinities. The DFT-calculated magnetic parameters of the low-energy conformers reproduce best the experimental electron paramagnetic resonance parameters measured in aqueous solutions for trans- and cis-Cu(Gln)2 conformers having two oxygen atoms (either from Gln or water molecules) at the apical positions, and Cu(His)(Gln) conformers having His in the histaminate-like mode with an apically placed carboxylato oxygen atom. The predicted conformational flexibility of His‑copper(II)-amino acid compounds may be connected with their physiological abundance, and the role in copper(II) exchange reactions in blood plasma.


Periodontal ligament stem cell-mediated treatment for periodontitis in miniature swine.
Periodontitis is a periodontal tissue infectious disease and the most common cause for tooth loss in adults. It has been linked to many systemic disorders, such as coronary artery disease, stroke, and diabetes. At present, there is no ideal therapeutic approach to cure periodontitis and achieve optimal periodontal tissue regeneration. In this study, we explored the potential of using autologous periodontal ligament stem cells (PDLSCs) to treat periodontal defects in a porcine model of periodontitis. The periodontal lesion was generated in the first molars area of miniature pigs by the surgical removal of bone and subsequent silk ligament suture around the cervical portion of the tooth. Autologous PDLSCs were obtained from extracted teeth of the miniature pigs and then expanded ex vivo to enrich PDLSC numbers. When transplanted into the surgically created periodontal defect areas, PDLSCs were capable of regenerating periodontal tissues, leading to a favorable treatment for periodontitis. This study demonstrates the feasibility of using stem cell-mediated tissue engineering to treat periodontal diseases.


Metabolic adverse events associated with systemic corticosteroid therapy-a systematic review and meta-analysis.
To assess the risk of new-onset or worsening hyperglycaemia, hypertension, weight gain and hyperlipidaemia with systemic corticosteroid therapy (CST) as reported in published randomised control trial (RCT) studies. Literature search using MEDLINE, EMBASE, Cochrane library, Web of Science and Scopus STUDY ELIGIBILITY CRITERIA: Published articles on results of RCT with a systemic CST arm with numerical data presented on adverse effect (AE). Reports of hyperglycaemia, hypertension, weight gain and hyperlipidaemia associated with systemic CST in patients or healthy volunteer's ≥17 years of age. Risk of bias tool, assessment at the level of AE and key study characteristics. A total of 5446 articles were screened to include 118 studies with 152 systemic CST arms (total participants=17 113 among which 8569 participants treated with CST). Pooled prevalence of hyperglycaemia in the CST arms within the studies was 10% (95% CI 7% to 14%), with the highest prevalence in respiratory illnesses at 22% (95% CI 9% to 35%). Pooled prevalence of severe hyperglycaemia, hypertension, weight gain and hyperlipidaemia within the corticosteroid arms was 5% (95% CI 2% to 9%), 6% (95% CI 4% to 8%), 13% (95% CI 8% to 18%), 8% (95% CI 4% to 17%), respectively. CST was significantly associated hyperglycaemia, hypertension and weight gain as noted in double-blinded placebo-controlled parallel-arms studies: OR of 2.13 (95% CI 1.66 to 2.72), 1.68 (95% CI 0.96 to 2.95) and 5.20 (95% CI 2.10 to 12.90), respectively. Intravenous therapy posed higher risk than oral therapy: OR of 2.39 (95% CI 1.16 to 4.91). There was significant heterogeneity in the AE definitions and quality of AE reporting in the primary studies and patient populations in the studies. The impact of cumulative dose effect on incidental AE could not be calculated. Systemic CST use is associated with increased risk of metabolic AEs, which differs for each disease group and route of administration. CRD42020161270.


Nonlinear optical imaging and spectral-lifetime computational analysis of endogenous and exogenous fluorophores in breast cancer.
Multiphoton laser scanning microscopy (MPLSM) utilizing techniques such as multiphoton excitation (MPE), second harmonic generation (SHG), and multiphoton fluorescence lifetime imaging and spectral lifetime imaging (FLIM and SLIM, respectively) are greatly expanding the degree of information obtainable with optical imaging in biomedical research. The application of these nonlinear optical approaches to the study of breast cancer holds particular promise. These noninvasive, multidimensional techniques are well suited to image exogenous fluorophores that allow relevant questions regarding protein localization and signaling to be addressed both in vivo and in vitro. Furthermore, MPLSM imaging of endogenous signals from collagen and fluorophores such as nicotinamide adenine dinucleotide (NADH) or flavin adenine dinucleotide (FAD), address important questions regarding the tumor-stromal interaction and the physiologic state of the cell. We demonstrate the utility of multimodal MPE/SHG/FLIM for imaging both exogenous and/or endogenous fluorophores in mammary tumors or relevant 3-D systems. Using SLIM, we present a method for imaging and differentiating signals from multiple fluorophores that can have overlapping spectra via SLIM Plotter-a computational tool for visualizing and analyzing large spectral-lifetime data sets.


Cloning of human Bcl-2 homologue: inflammatory cytokines induce human A1 in cultured endothelial cells.
Bcl-2 is an intracellular membrane-associated protein that functions to block programmed cell death. Despite recurrent exposure to cellular toxins from the circulation and tissue, endothelial cells are remarkably resistant to cell death. Because Bcl-2 protein levels are low or undetectable in endothelial cells, we postulated that other members of the growing Bcl-2 family would be present in endothelial cells to provide protection against apoptosis. Degenerate primers to two conserved regions of the Bcl-2 family were used to amplify potential homologues in endothelial cells. This strategy resulted in the isolation of a human Bcl-2 homologue related to murine Al, a recently identified member of this family. We show here that, in endothelial cells, human Al is rapidly inducible by phorbol ester and the inflammatory cytokines, tumor necrosis factor-alpha and interleukin-1beta, but not by the growth factors, basic fibroblast growth factor or vascular endothelial growth factor. Al is the only known Bcl-2 family member that is inducible by inflammatory cytokines, suggesting that it may play a protective role during inflammation. Additionally, vascular smooth muscle cells and various nonhematopoietic tissues express human Al, indicating that human Al is a widely expressed Bcl-2 homologue.


Effect of the length of the cycle on biodegradable polymer production and microbial community selection in a sequencing batch reactor.
The effect of the length of the cycle on the enrichment and selection of mixed cultures in sequencing batch reactors (SBRs) has been studied, with the aim of biodegradable polymers (namely, polyhydroxyalkanoates (PHAs)) production from organic wastes. At a fixed feed concentration (20 gCOD/L) and organic loading rate (20 gCOD/L/day), the SBR was operated at different lengths of the cycle, in the range 1-8 h. Process performance was measured by considering the rates and yields of polymer storage and of the competing phenomenon of growth. The selected biomass was enriched with microorganisms that were able to store PHAs at high rates and yields only when the length of the cycle was 2 or 4 h, even though in these conditions the process was unstable. On the other hand, when the length of the cycle was 1 or 8 h, the dynamic response of the selected microorganisms was dominated by growth. The best process performance was characterized by storage rates in the range 500-600 mgCOD/gCOD/h and storage yields of 0.45-0.55 COD/COD. The corresponding productivity of the process was in the range 0.25-0.30 gPHA/L/h, the highest values obtained until now for mixed cultures. The microbial composition of the selected biomasses was analyzed through denaturing gradient gel electrophoresis (DGGE) and reverse-transcriptase denaturing gradient gel electrophoresis (RT-DGGE). The instability of the runs characterized by high storage rate was associated with a higher microbial heterogeneity compared to the runs with a stable growth response.


Quantification method of size distribution of polysilicic acid in supersaturated silicic acid solution.
In order to estimate the absolute size distribution of polysilicic acid particles in geothermal waters, the distribution coefficient (K(av)) of gel permeation chromatography (GPC) for polysilicic acid particles and the hydrodynamic radius for the same polysilicic acid particles from Dynamic Light Scattering (DLS) are combined to quantify the particle size. From the combination, a quantitative relationship between the K(av) from GPC and the hydrodynamic radius for polysilicic acid from DLS was built up. Using this relationship, the change in particle size of polysilicic acid formed during the polymerization of silicic acid at pH 8 and 9 (initial silicic acid concentration: 800 ppm as SiO2) was examined. The result showed that polysilicic acid grew to 500 and 1000 nm by 5 h at pH 9 and 8, respectively. It was found that aluminum affects the growth of polysilicic acid particles, and that the effect depends on the pH. The proposed method in this study has been proved to be valid to measure the size of polysilicic acid during the polymerization of silicic acid in solutions with relatively low silicic acid concentration, such as geothermal water.


Enzyme linked direct antiglobulin tests in patients with autoimmune haemolysis.
Enzyme linked and agglutination direct antiglobulin tests were carried out on blood samples from 219 patients suspected of having autoimmune haemolysis. The enzyme linked tests were more sensitive: they could detect the small amounts of IgG, IgA, and IgM which are normally present on red cells and showed increased amounts of cell bound immunoglobulins in patients with Coombs test negative autoimmune haemolysis. Many patients had immunoglobulins of more than one class bound to their red cells; considering the degree of haemolysis in individual patients, it appeared that the different immunoglobulin classes acted synergistically in effecting red cell destruction, even in amounts too small to be detected by the agglutination tests. In patients with cold reacting autoantibodies and complement coating of the red cells active haemolysis was found (with one exception) where IgM was detected on the cells by the enzyme linked method. Elution studies indicated that immunoglobulins detected just by the enzyme linked techniques were red cell antibodies. Both enzyme linked and agglutination tests were negative in 66 patients: 61 of these had no evidence of haemolysis, and in the other five the haemolysis was not autoimmune in origin.


Separable Markov random field model and its applications in low level vision.
This brief proposes a continuously-valued Markov random field (MRF) model with separable filter bank, denoted as MRFSepa, which significantly reduces the computational complexity in the MRF modeling. In this framework, we design a novel gradient-based discriminative learning method to learn the potential functions and separable filter banks. We learn MRFSepa models with 2-D and 3-D separable filter banks for the applications of gray-scale/color image denoising and color image demosaicing. By implementing MRFSepa model on graphics processing unit, we achieve real-time image denoising and fast image demosaicing with high-quality results.


Semaphorin 7A is protective during inflammatory peritonitis through integrin receptor signaling.
The study explores the role of endothelial Semaphorin 7A (SEMA7A) in inflammatory processes. SEMA7A is known for enhancing inflammation during tissue hypoxia and exhibiting anti-inflammatory properties in the intestinal system during colitis. This research extends the understanding of SEMA7A's function by examining its role in inflammatory peritonitis and intestinal inflammation. The research involved inducing peritonitis in SEMA7A knockout (SEMA7A-/-) and wild-type (WT) animals through Zymosan A (ZyA) injection. The inflammatory response was assessed by measuring cell count and cytokine release. In parallel, the study investigated the expression of SEMA7A in intestinal epithelial cells under inflammatory stimuli and its impact on interleukin 10 (IL-10) production using an in vitro co-culture model of monocytes and epithelial cells. Additionally, the distribution of SEMA7A target receptors, particularly ITGAV/ITGB1 (CD51/CD29), was analyzed in WT animals. The results revealed that SEMA7A-/- animals exhibited increased inflammatory peritonitis compared to the WT animals. Inflammatory conditions in intestinal epithelial cells led to the induction of SEMA7A. The co-culture experiments demonstrated that SEMA7A induced IL-10 production, which depended on integrin receptors and was independent of PLXNC1 expression. Furthermore, ITGAV/ITGB1 emerged as the predominant SEMA7A receptor in the intestinal area of WT animals. These findings underscore the multifaceted role of SEMA7A in inflammatory processes. The differential responses in peritonitis and intestinal inflammation suggest that SEMA7A's function is significantly influenced by the expression and distribution of its target receptors within different organ systems. The study highlights the complex and context-dependent nature of SEMA7A in mediating inflammatory responses.


Stepping in Place While Voluntarily Turning Around Produces a Long-Lasting Posteffect Consisting in Inadvertent Turning While Stepping Eyes Closed.
Training subjects to step in place on a rotating platform while maintaining a fixed body orientation in space produces a posteffect consisting in inadvertent turning around while stepping in place eyes closed (podokinetic after-rotation, PKAR). We tested the hypothesis that voluntary turning around while stepping in place also produces a posteffect similar to PKAR. Sixteen subjects performed 12 min of voluntary turning while stepping around their vertical axis eyes closed and 12 min of stepping in place eyes open on the center of a platform rotating at 60°/s (pretests). Then, subjects continued stepping in place eyes closed for at least 10 min (posteffect). We recorded the positions of markers fixed to head, shoulder, and feet. The posteffect of voluntary turning shared all features of PKAR. Time decay of angular velocity, stepping cadence, head acceleration, and ratio of angular velocity after to angular velocity before were similar between both protocols. Both postrotations took place inadvertently. The posteffects are possibly dependent on the repeated voluntary contraction of leg and foot intrarotating pelvic muscles that rotate the trunk over the stance foot, a synergy common to both protocols. We propose that stepping in place and voluntary turning can be a scheme ancillary to the rotating platform for training body segment coordination in patients with impairment of turning synergies of various origin.


Humoral immune response in human tuberculosis: immunoglobulins G, A, and M directed against the purified P32 protein antigen of Mycobacterium bovis bacillus Calmette-Guérin.
The P32 protein antigen of Mycobacterium bovis BCG, identified as antigen 85A in the BCG reference system, was used to investigate the humoral immune response in human tuberculosis (TB). Immunoglobulin G (IgG), IgA, and IgM directed against P32 were measured by an enzyme-linked immunosorbent assay. Mean IgG and IgA antibody levels differed significantly (P less than 0.001) between active-TB patients (50 untreated and 52 treated) and healthy control subjects (111 unvaccinated tuberculin negative, 38 unvaccinated tuberculin positive, and 72 recently BCG vaccinated). Mean IgG antibody levels, but not mean IgA antibody levels, were higher (P less than 0.05) in patients with positive microscopic examination for acid-fast bacilli than in patients with negative microscopic examination. A positive relation was found between mean levels and the extent of disease. There was no difference in mean IgM antibody levels between patients and controls. By setting the upper normal limit at the 95th percentile of the 221 healthy subjects, the sensitivities were 46% in untreated and 63% in treated patients for IgG and 30 and 50%, respectively, for IgA. Of the untreated patients, 56% were positive for either IgG or IgA antibodies. Among the untreated patients with negative direct smear, 35% were positive for IgG and 24% were positive for IgA. When both immunoglobulin classes were combined, the serological test was positive in 47% of those patients. Neither naturally acquired tuberculin hypersensitivity nor BCG vaccination affected positivity frequencies in healthy subjects. Only active TB seemed to induce significant anti-P32 antibody levels and to be associated with positivity. A serological test with P32 as the antigen might therefore be helpful for the rapid diagnosis of TB.


Research supervision.
Nurses are expected to base their practice on research, and to carry out research themselves. Whereas research supervision is useful to the experienced researcher, it is vital to the novice. It is surprising, therefore, that there is little published work on research supervision. Practical advice based on first-hand experience is the focus for most work and, ironically, very little of that available is research based. This literature review is categorised into three main sections: the supervision relationship, good practice in supervision, and the context of supervision. A review is necessary to address what information is known, to provide guidance both to supervisors and supervisees, and to highlight the need for further work. The little research evidence that is available suggests a way forward based on practical experience of both supervising and being supervised. The implications of this are briefly outlined.


Transitions Study of predictors of illness progression in young people with mental ill health: study methodology.
An estimated 75% of mental disorders begin before the age of 24 and approximately 25% of 13-24-year-olds are affected by mental disorders at any one time. To better understand and ideally prevent the onset of post-pubertal mental disorders, a clinical staging model has been proposed that provides a longitudinal perspective of illness development. This heuristic model takes account of the differential effects of both genetic and environmental risk factors, as well as markers relevant to the stage of illness, course or prognosis. The aim of the Transitions Study is to test empirically the assumptions that underpin the clinical staging model. Additionally, it will permit investigation of a range of psychological, social and genetic markers in terms of their capacity to define current clinical stage or predict transition from less severe or enduring to more severe and persistent stages of mental disorder. This paper describes the study methodology, which involves a longitudinal cohort design implemented within four headspace youth mental health services in Australia. Participants are young people aged 12-25 years who have sought help at headspace and consented to complete a comprehensive assessment of clinical state and psychosocial risk factors. A total of 802 young people (66% female) completed baseline assessments. Annual follow-up assessments have commenced. The results of this study may have implications for the way mental disorders are diagnosed and treated, and progress our understanding of the pathophysiologies of complex mental disorders by identifying genetic or psychosocial markers of illness stage or progression.


Phase I investigator-initiated study of the safety of MTC001 in patients with chronic ischemic heart failure.
: Heart failure (HF) is a global pandemic most commonly caused by coronary artery disease. Despite coronary revascularization, the infarcted myocardium can develop into an irreversible scar toward chronic ischemic HF. This is due to the limited regenerative capacity of the adult human heart. Recently, the vascular cell adhesion molecule 1 positive cardiac fibroblast (VCF) has been shown to directly improve cardiac contractility in addition to promoting myocardial growth in preclinical studies. This clinical trial aims to explore the safety and, in part, the efficacy of autologous VCF therapy for chronic ischemic HF. : This first-in-human trial is an open-label, single-arm, phase 1 study conducted at a single center. This study will include 6 patients with chronic ischemic HF in stage C and NYHA class II or III despite receiving the standard of care, including coronary revascularization. Participants will undergo cardiac biopsy to manufacture autologous VCFs expressing CD90 and CD106. Under electro-anatomical mapping guidance, participants will receive a transendocardial injection of VCF in a modified 3 + 3 design. The first 3 patients will receive a standard dose (2 × 107 cells) of VCF with a 4-week interval for safety assessment before subsequent enrollment. In the absence of safety issues, the final 3 patients will receive the standard dose of VCF without a 4-week interval. In the presence of safety issues, the final 3 patients will receive a reduced dose (1.5 × 107 cells) of VCF with the 4-week interval. This is the first clinical study of cardiac regeneration using VCFs for the treatment of chronic ischemic HF. The study results will contribute to the development of a minimally invasive cell therapy for patients with HF failed by the standard of care. This study was registered with the Japan Registry of Clinical Trials (jRCT2033210078).


Longitudinal associations of blood markers of insulin and glucose metabolism and cancer mortality in the third National Health and Nutrition Examination Survey.
Insulin and glucose may influence cancer mortality via their proliferative and anti-apoptotic properties. Using longitudinal data from the nationally representative Third National Health and Nutrition Examination Survey (NHANES III; 1988-1994), with an average follow-up of 8.5 years to death, we evaluated markers of glucose and insulin metabolism, with cancer mortality, ascertained using death certificates or the National Death Index. Plasma glucose, insulin, C-peptide, and lipid concentrations were measured. Anthropometrics, lifestyle, medical, and demographic information was obtained during in-person interviews. After adjusting for age, race, sex, smoking status, physical activity, and body mass index, for every 50 mg/dl increase in plasma glucose, there was a 22% increased risk of overall cancer mortality. Insulin resistance was associated with a 41% (95% confidence interval (CI) (1.07-1.87; p = 0.01) increased risk of overall cancer mortality. These associations were stronger after excluding lung cancer deaths for insulin-resistant individuals (HR: 1.67; 95% CI: 1.15-2.42; p = 0.01), specifically among those with lower levels of physical activity (HR: 2.06; 95% CI: 1.4-3.0; p = 0.0001). Similar associations were observed for other blood markers of glucose and insulin, albeit not statistically significant. In conclusion, hyperglycemia and insulin resistance may be 'high-risk' conditions for cancer mortality. Managing these conditions may be effective cancer control tools.


Dairy farmer and farm staff attitudes and perceptions regarding daily milk allowance to calves.
The benefits of feeding calves more milk are increasingly being recognized by dairy farmers. However, most producers have still not implemented higher feeding plans. The aim of the present study was to gain a deeper understanding of farmer and farm staff attitudes, and the perceptions and factors considered in their decision-making regarding daily milk allowances. We collected data through focus group interviews with dairy farmers, farm managers, and calf-care workers who were selected using purposive and snowball sampling. In total, 40 persons (24 women and 16 men) joined a focus group interview (6 in all, each with 5-8 participants). Interviews were recorded, and recordings were transcribed and analyzed thematically. Participants had contrasting opinions about the minimum, maximum, and recommended daily milk allowances to their calves. Their suggested lowest daily milk allowance to sustain animal welfare ranged from 4 to 8-10 L and the maximum allowance from 6 to 15 L. We found that farmers' and farm staff's choices and recommendations of milk-feeding protocols were influenced by a large number of factors that could be grouped into 4 themes: (1) Life beyond work, (2) Farm facilities and equipment, (3) Care of the calves, and (4) Profitability and production. Participants' considerations were similar and aimed to maximize daily milk allowance based on farm conditions. However, the allowances they described as optimal for their calves often differed from what they considered practically feasible. We found that the care of the calves and the well-being of the owners and the staff was central in the participants' decision-making, but that this care perspective was challenged by the social and economic sustainability of the farm. Most participants fed their calves twice daily and did not think that increasing that number would be practically feasible. Our results indicate that the participants' viewpoints regarding calves were important for their decision-making about milk allowances. We suggest that a more holistic perspective should be used when advising farmers about milk allowances, putting particular emphasis on the caring and social sustainability aspects of the individual farm.


Roadmap for C9ORF72 in Frontotemporal Dementia and Amyotrophic Lateral Sclerosis: Report on the C9ORF72 FTD/ALS Summit.
A summit held March 2023 in Scottsdale, Arizona (USA) focused on the intronic hexanucleotide expansion in the C9ORF72 gene and its relevance in frontotemporal dementia (FTD) and amyotrophic lateral sclerosis (ALS; C9ORF72-FTD/ALS). The goal of this summit was to connect basic scientists, clinical researchers, drug developers, and individuals affected by C9ORF72-FTD/ALS to evaluate how collaborative efforts across the FTD-ALS disease spectrum might break down existing disease silos. Presentations and discussions covered recent discoveries in C9ORF72-FTD/ALS disease mechanisms, availability of disease biomarkers and recent advances in therapeutic development, and clinical trial design for prevention and treatment for individuals affected by C9ORF72-FTD/ALS and asymptomatic pathological expansion carriers. The C9ORF72-associated hexanucleotide repeat expansion is an important locus for both ALS and FTD. C9ORF72-FTD/ALS may be characterized by loss of function of the C9ORF72 protein and toxic gain of functions caused by both dipeptide repeat (DPR) proteins and hexanucleotide repeat RNA. C9ORF72-FTD/ALS therapeutic strategies discussed at the summit included the use of antisense oligonucleotides, adeno-associated virus (AAV)-mediated gene silencing and gene delivery, and engineered small molecules targeting RNA structures associated with the C9ORF72 expansion. Neurofilament light chain, DPR proteins, and transactive response (TAR) DNA-binding protein 43 (TDP-43)-associated molecular changes were presented as biomarker candidates. Similarly, brain imaging modalities (i.e., magnetic resonance imaging [MRI] and positron emission tomography [PET]) measuring structural, functional, and metabolic changes were discussed as important tools to monitor individuals affected with C9ORF72-FTD/ALS, at both pre-symptomatic and symptomatic disease stages. Finally, summit attendees evaluated current clinical trial designs available for FTD or ALS patients and concluded that therapeutics relevant to FTD/ALS patients, such as those specifically targeting C9ORF72, may need to be tested with composite endpoints covering clinical symptoms of both FTD and ALS. The latter will require novel clinical trial designs to be inclusive of all patient subgroups spanning the FTD/ALS spectrum.


Trends in Premature Mortality From Acute Myocardial Infarction in the United States, 1999 to 2019.
Background Evaluating premature (<65 years of age) mortality because of acute myocardial infarction (AMI) by demographic and regional characteristics may inform public health interventions. Methods and Results We used the Centers for Disease Control and Prevention's WONDER (Wide-Ranging Online Data for Epidemiologic Research) death certificate database to examine premature (<65 years of age) age-adjusted AMI mortality rates per 100 000 and average annual percentage change from 1999 to 2019. Overall, the age-adjusted AMI mortality rate was 13.4 (95% CI, 13.3-13.5). Middle-aged adults, men, non-Hispanic Black adults, and rural counties had higher mortality than young adults, women, NH White adults, and urban counties, respectively. Between 1999 and 2019, the age-adjusted AMI mortality rate decreased at an average annual percentage change of -3.4 per year (95% CI, -3.6 to -3.3), with the average annual percentage change showing higher decline in age-adjusted AMI mortality rates among large (-4.2 per year [95% CI, -4.4 to -4.0]), and medium/small metros (-3.3 per year [95% CI, -3.5 to -3.1]) than rural counties (-2.4 per year [95% CI, -2.8 to -1.9]). Age-adjusted AMI mortality rates >90th percentile were distributed in the Southern states, and those with mortality <10th percentile were clustered in the Western and Northeastern states. After an initial decline between 1999 and 2011 (-4.3 per year [95% CI, -4.6 to -4.1]), the average annual percentage change showed deceleration in mortality since 2011 (-2.1 per year [95% CI, -2.4 to -1.8]). These trends were consistent across both sexes, all ethnicities and races, and urban/rural counties. Conclusions During the past 20 years, decline in premature AMI mortality has slowed down in the United States since 2011, with considerable heterogeneity across demographic groups, states, and urbanicity. Systemic efforts are mandated to address cardiovascular health disparities and outcomes among nonelderly adults.


Sudden infant death syndrome, bedsharing, parental weight, and age at death.
To assess the role of parental bedsharing in sudden infant death syndrome (SIDS)-like deaths, this study examines the hypothesis that, compared with other SIDS cases, the age distribution of deaths associated with bedsharing should be lower in younger, less vigorous infants. For 84 SIDS cases in Cleveland, Ohio, 1992 to 1996, age at death, maternal weight, and other risk factors for SIDS were compared for cases grouped according to bedsharing status. Mean ages at death were 9.1 weeks for 30 bedsharing and 12.7 for 54 nonbedsharing cases, counting 10 with missing information as nonbedsharing. Mean pregravid weights of bedsharing mothers exceeded those of nonbedsharing mothers (84.1 vs 67.0 kg). Mean ages at death for nonbedsharing infants, bedsharing infants of smaller mothers, and bedsharing infants of larger mothers were 12.7, 10.3, and 7.6 weeks, respectively. Large maternal size did not affect age at death in the absence of bedsharing. By demonstrating that among an urban population at high risk for SIDS, bedsharing is strongly associated with a younger age at death, independent of any other factors, this study provides evidence of a relationship between some SIDS-like deaths and parent-infant bedsharing, particularly if the parent is large.


Usefulness of a decision tree model for the analysis of adverse drug reactions: Evaluation of a risk prediction model of vancomycin-associated nephrotoxicity constructed using a data mining procedure.
Several publications concerning decision tree (DT) analysis in medical fields have recently demonstrated its usefulness for defining prognostic factors in various diseases. However, there are minimal reports on the predictors of adverse drug reactions. We attempted to use DT analysis to discover combinations of multiple risk factors that would increase the risk of nephrotoxicity associated with vancomycin (VCM). To demonstrate the usefulness of DT analysis, we compared its predictive performance with that of multiple logistic regression analysis. A single-centre, retrospective study was conducted at Hokkaido University Hospital. A total of 592 patients, who received intravenous administrations of VCM between November 2011 and April 2016, were enrolled. Nephrotoxicity was defined as an increase in serum creatinine of ≥0.5 mg/dL or a ≥50% increase in serum creatinine from the baseline. Risk factors for VCM nephrotoxicity were extracted from previous reports. In the DT analysis, a chi-squared automatic interaction detection algorithm was constructed. For evaluating the established algorithms, a 10-fold cross validation method was adopted to calculate the misclassification risk of the model. Moreover, to compare the accuracy of the DT analysis, multiple logistic regression analysis was conducted. Eighty-seven (14.7%) patients developed nephrotoxicity. A VCM trough concentration of ≥15.0 mg/L, concomitant medication (vasopressor drugs and furosemide), and a duration of therapy ≥14 days were extracted to build the DT model, in which the patients were divided into 6 subgroups based on variable rates of nephrotoxicity, ranging from 4.6 to 69.6%. The predictive accuracies of the DT and logistic regression models were similar (87.3%, respectively), indicating that they were accurate. This study suggests the usefulness of DT models for the evaluation of adverse drug reactions.


Principles in laboratory animal research for experimental purposes.
The present work contains information about proper husbandry and care of laboratory animals, microbiological monitoring of their health and protecting them against suffering and distress. The author also gives some advice on the improvement and unification of experimental research results through the standardisation of laboratory animals used for the experiments as well as imposing proper conditions for animal husbandry.


Characterization of reproductive steroid receptors and response to estrogen in a rat serotonergic cell line.
Study of the cellular and molecular consequences of steroid hormone action in the serotonin neural system will provide new avenues for pharmacotherapeutic intervention in mental illness related to reproductive function. However, it is difficult to probe intracellular mechanisms with whole animal models. We sought the steroid receptor compliment and estrogen response of two rat serotonin cell lines in order to determine if they could be of future assistance in this matter. Immunohistochemistry with a panel of antibodies, RT-PCR and a serotonin ELISA were utilized to characterize the RN46A-V1 cells (herein called RN46A), and the subclone RN46A-B14 (herein called B14) that is stably transfected with brain derived neurotrophic factor (BDNF). RN46A and B14 cells express estrogen receptor beta (ERbeta), androgen receptors (AR) and nuclear factor kappa B (NFkappaB) but not estrogen receptor alpha (ERalpha) or progestin receptors (PR). RT-PCR confirmed the presence of ERbeta and the absence of ERalpha and PR in both cell lines. B14 cells contain more immunodetectable BDNF and serotonin than the RN46A parent line. In addition, immunofluorescence for the serotonin reuptake transporter (SERT) was observed in the cell body region of undifferentiated B14 cells. After differentiation at a nonpermissive temperature, SERT immunostaining was observed in both the cell body region and along the extent of the axons. Serotonin content as determined by ELISA was higher in B14 than RN46A cells. Estrogen (0.1 and 1.0 nM) stimulated serotonin in the B14 cells in serum free medium. In summary, the RN46A cells and the B14 subclone contain the same compliment of nuclear steroid receptors as rat raphe serotonin neurons and thus may provide a convenient in vitro model for study of intracellular mechanisms of action of steroid hormones in the context of a serotonin neuron.


Comparing evolutionary strategies on a biobjective cultural algorithm.
Evolutionary algorithms have been widely used to solve large and complex optimisation problems. Cultural algorithms (CAs) are evolutionary algorithms that have been used to solve both single and, to a less extent, multiobjective optimisation problems. In order to solve these optimisation problems, CAs make use of different strategies such as normative knowledge, historical knowledge, circumstantial knowledge, and among others. In this paper we present a comparison among CAs that make use of different evolutionary strategies; the first one implements a historical knowledge, the second one considers a circumstantial knowledge, and the third one implements a normative knowledge. These CAs are applied on a biobjective uncapacitated facility location problem (BOUFLP), the biobjective version of the well-known uncapacitated facility location problem. To the best of our knowledge, only few articles have applied evolutionary multiobjective algorithms on the BOUFLP and none of those has focused on the impact of the evolutionary strategy on the algorithm performance. Our biobjective cultural algorithm, called BOCA, obtains important improvements when compared to other well-known evolutionary biobjective optimisation algorithms such as PAES and NSGA-II. The conflicting objective functions considered in this study are cost minimisation and coverage maximisation. Solutions obtained by each algorithm are compared using a hypervolume S metric.


Controllable synthesis of hierarchical porous Fe3O4 particles mediated by poly(diallyldimethylammonium chloride) and their application in arsenic removal.
Hierarchical porous Fe3O4 particles with tunable grain size were synthesized based on a facile poly (diallyldimethylammonium chloride) (PDDA)-modulated solvothermal method. The products were characterized with scanning electron microscopy (SEM) and transmission electron microscopy (TEM), X-ray photoelectron spectroscopy (XPS), Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD), N2 adsorption-desorption technique, vibrating sample magnetometer (VSM), and dynamic light scattering (DLS). The results show that increasing the PDDA dosage decrease the grain size and particle size, which increased the particle porosity and enhanced the surface area from 7.05 to 32.75 m(2) g(-1). Possible mechanism can be ascribed to the PDDA function on capping the crystal surface and promoting the viscosity of reaction medium to mediate the growth and assembly of grain. Furthermore, the arsenic adsorption application of the as-obtained Fe3O4 samples was investigated and the adsorption mechanism was proposed. High magnetic Fe3O4 particles with increased surface area display improved arsenic adsorption performance, superior efficiency in low-level arsenic removal, high desorption efficiency, and satisfactory magnetic recyclability, which are very promising compared with commercial Fe3O4 particles.


Differential expression of extracellular matrix‑related genes DCN, EPHA4, FN1, SPARC, SPON2 and SPP1 in colorectal carcinogenesis.
During colorectal carcinogenesis, a spectrum of lesions is formed with different malignant potentials and occasionally with ambiguous morphologic features. To identify novel biomarkers, we previously performed a bioinformatics study to detect differentially expressed genes between colorectal adenoma (CRA) and colorectal carcinoma (CRC). The present study validated the diagnostic and prognostic significance of 6 components of the extracellular matrix (ECM) that were identified in the previous bioinformatics analysis [decorin (DCN), erythropoietin‑producing hepatoma receptor A4 (EPHA4), fibronectin 1 (FN1), secreted protein acidic and cysteine rich (SPARC), spondin 2 (SPON2) and secreted phosphoprotein 1 (SPP1)], by analyzing their gene and protein expression levels in all samples. A total of 60 formalin‑fixed paraffin‑embedded biopsy samples were included in the present study, from 40 patients with CRA, malignant polyps and CRC with and without lymph node metastases. The expression of the ECM‑related genes was evaluated on the mRNA level using reverse transcription‑quantitative PCR (RT‑qPCR) and on the protein level using immunohistochemistry. RT‑qPCR results revealed differential expression among the groups for all genes, except for EPHA4. Their expression proportionally increased with the level of malignancy. Among them, SPARC, SPON2 and SPP1 were differentially expressed between CRA and malignant polyps. Immunohistochemistry analysis revealed two patterns: Positive staining of SPON2 and SPP1 in epithelial cells of healthy mucosa and dysplastic glands of CRA and CRC, and positive staining of DCN and SPARC in the lamina propria in healthy mucosa and stroma of CRA and CRC. The intensity of staining increased with the severity of lesions. The present results suggested that ECM‑related proteins may have an important role in the development of CRC, with a possible diagnostic use for differentiation among various lesions, such as between CRA and malignant polyps. However, no significant potential was detected for these genes to predict lymph node metastases in CRC.


Multiple osteosclerotic lesions in an Iron Age skull from Switzerland (320-250 BC)--an unusual case.
The single Hochdorf burial was found in 1887 during construction work in the Canton of Lucerne, Switzerland. It dates from between 320 and 250 BC. The calvarium, the left half of the pelvis and the left femur were preserved. The finding shows an unusual bony alteration of the skull. The aim of this study was to obtain a differential diagnosis and to examine the skull using various methods. Sex and age were determined anthropologically. Radiological examinations were performed with plain X-ray imaging and a multislice computed tomography (CT) scanner. For histological analysis, samples of the lesion were taken. The pathological processing included staining after fixation, decalcification, and paraffin embedding. Hard-cut sections were also prepared. The individual was female. The age at death was between 30 and 50 years. There is an intensely calcified bone proliferation at the right side of the os frontalis. Plain X-ray and CT imaging showed a large sclerotic lesion in the area of the right temple with a partly bulging appearance. The inner boundary of the lesion shows multi-edged irregularities. There is a diffuse thickening of the right side. In the left skull vault, there is a mix of sclerotic areas and areas which appear to be normal with a clear differentiation between tabula interna, diploë and tabula externa. Histology showed mature organised bone tissue. Radiological and histological findings favour a benign condition. Differential diagnoses comprise osteomas which may occur, for example, in the setting of hereditary adenomatous polyposis coli related to Gardner syndrome.


Using infection prevalence, seroprevalence and case report data to estimate chlamydial infection incidence.
To measure the effectiveness of chlamydia control strategies, we must estimate infection incidence over time. Available data, including survey-based infection prevalence and case reports, have limitations as proxies for infection incidence. We therefore developed a novel method for estimating chlamydial incidence. We linked a susceptible infectious mathematical model to serodynamics data from the National Health and Nutritional Examination Survey, as well as to annual case reports. We created four iterations of this model, varying assumptions about how the method of infection clearance (via treatment seeking, routine screening or natural clearance) relates to long-term seropositivity. Using these models, we estimated annual infection incidence for women aged 18-24 and 25-37 years in 2014. To assess model plausibility, we also estimated natural clearance for the same groups. Of the four models we analysed, the model that best explained the empirical data was the one in which longer-lasting infections, natural clearance and symptomatic infections all increased the probability of long-term seroconversion. Using this model, we estimated 5910 (quartile (Q)1, 5330; Q3, 6500) incident infections per 100 000 women aged 18-24 years and 2790 (Q1, 2500; Q3, 3090) incident infections per 100 000 women aged 25-37 years in 2014. Furthermore, we estimated that natural clearance rates increased with age. Our method can be used to estimate the number of chlamydia infections each year, and thus whether infection incidence increases or decreases over time and after policy changes. Furthermore, our results suggest that clearance via medical intervention may lead to short-term or no seroconversion, and the duration of untreated chlamydial infection may vary with age, underlining the complexity of chlamydial infection dynamics.


An open-label study on the effect of flax seed powder (Linum usitatissimum) supplementation in the management of diabetes mellitus.
Diabetes mellitus is characterized by hyperglycemia and associated with aberrations in the metabolism of carbohydrate, protein, and lipid that result in development of secondary complications. Extensive studies have indicated that nutritional therapy plays a pivotal role in the controlling or postponing of development of these secondary complications. Several functional foods have been shown to possess hypoglycemic and hypolipidemic properties. Flax seed (FS) is a functional food that is rich in omega 3 fatty acids and antioxidants and is low in carbohydrates. In exploratory studies, FS was incorporated in recipes, which resulted in a reduction in the glycemic index of the food items. These observations prompted us to investigate the efficacy of FS supplementation in type 2 diabetics (n = 29). Subjects were assigned to the experimental (n = 18) or the control group (n = 11) on the basis of their desire to participate in the study. The experimental group's diet was supplemented daily with 10 g of FS powder for a period of 1 month. The control group received no supplementation or placebo. During the study, diet and drug intake of the subjects remained unaltered. The efficacy of supplementation with FS was evaluated through a battery of clinico-biochemical parameters. Supplementation with FS reduced fasting blood glucose by 19.7% and glycated hemoglobin by 15.6%. A favorable reduction in total cholesterol (14.3%), triglycerides (17.5%), low-density lipoprotein cholesterol (21.8%), and apolipoprotein B and an increase in high-density lipoprotein cholesterol (11.9%) were also noticed. These observations suggest the therapeutic potential of FS in the management of diabetes mellitus.


Eight of fourteen gvp genes are sufficient for formation of gas vesicles in halophilic archaea.
The minimal number of genes required for the formation of gas vesicles in halophilic archaea has been determined. Single genes of the 14 gvp genes present in the p-vac region on plasmid pHH1 of Halobacterium salinarum (p-gvpACNO and p-gvpDEFGHIJKLM) were deleted, and the remaining genes were tested for the formation of gas vesicles in Haloferax volcanii transformants. The deletion of six gvp genes (p-gvpCN, p-gvpDE, and p-gvpHI) still enabled the production of gas vesicles in H. volcanii. The gas vesicles formed in some of these gvp gene deletion transformants were altered in shape (Delta I, Delta C) or strength (Delta H) but still functioned as flotation devices. A minimal p-vac region (minvac) containing the eight remaining genes (gvpFGJKLM-gvpAO) was constructed and tested for gas vesicle formation in H. volcanii. The minvac transformants did not form gas vesicles; however, minvac/gvpJKLM double transformants contained gas vesicles seen as light refractile bodies by phase-contrast microscopy. Transcript analyses demonstrated that minvac transformants synthesized regular amounts of gvpA mRNA, but the transcripts derived from gvpFGJKLM were mainly short and encompassed only gvpFG(J), suggesting that the gvpJKLM genes were not sufficiently expressed. Since gvpAO and gvpFGJKLM are the only gvp genes present in minvac/JKLM transformants containing gas vesicles, these gvp genes represent the minimal set required for gas vesicle formation in halophilic archaea. Homologs of six of these gvp genes are found in Anabaena flos-aquae, and homologs of all eight minimal halobacterial gvp genes are present in Bacillus megaterium and in the genome of Streptomyces coelicolor.


The hypolipidemic effect of locust bean gum food products in familial hypercholesterolemic adults and children.
Seventeen adults and 11 children, a group of 18 familial hypercholesterolemic (FHC) and 10 normal subjects, were fed products with and without locust bean gum (LBG) (8 to 30 g/day) to assess the hypolipidemic effect of LBG. Identical food products with and without LBG were consumed by two groups (A and B) of arbitrarily assigned patients using a cross-over design. Plasma cholesterol, low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol, very low-density lipoprotein cholesterol and triglycerides were measured at 2-wk intervals and compared to control feeding periods. In group A, FHC C decreased 10% and LDL-C 11%, normal subjects decreased C 6% and LDL-C 10% (p less than 0.001). In group B, FHC C decreased 17% and LDL-C 19%, normal subjects decreased cholesterol 11%, and LDL-C 6% (p less than 0.001). Cholesterol and LDL-C were lowered in FHC children in both groups. High-density lipoprotein/LDL ratios increased in both groups. The use of food products with LBG in children and adults is a unique approach to treating FHC. LBG food acceptance was good, and there were no significant side effects. LBG in food products appears to be an effective, safe approach to controlling hyperlipidemia.


Immune activation by a sterile aqueous extract of Cordyceps sinensis: mechanism of action.
Cordyceps sinensis is a fungus that has been used for over 2,000 years in China as a treatment for a variety of conditions including infectious diseases. The available evidence suggests a hypothesis that any efficacy of C. sinensis as an anti-infective therapeutic would be related to a role as an activator of innate immune responses. The objectives of this study were first to investigate the ability of C. sinensis to activate pro-inflammatory responses in macrophages in vitro and induce protective responses against intracellular pathogens in vivo, and second to characterize a method of action. We found that C. sinensis activates murine macrophages to produce a variety of pro-inflammatory cytokines. IFN-gamma synergizes with C. sinensis to amplify this response. Bacterial endotoxin contamination was ruled out as a potential artefact. The evidence presented in this study supports a hypothesis that C. sinensis activates macrophages by engaging Toll-like receptors and inducing mitogen-activated protein kinase (MAPK) pathways characteristic of inflammatory stimuli.


Reduced sequence variability on the Neo-Y chromosome of Drosophila americana americana.
Sex chromosomes are generally morphologically and functionally distinct, but the evolutionary forces that cause this differentiation are poorly understood. Drosophila americana americana was used in this study to examine one aspect of sex chromosome evolution, the degeneration of nonrecombining Y chromosomes. The primary X chromosome of D. a. americana is fused with a chromosomal element that was ancestrally an autosome, causing this homologous chromosomal pair to segregate with the sex chromosomes. Sequence variation at the Alcohol Dehydrogenase (Adh) gene was used to determine the pattern of nucleotide variation on the neo-sex chromosomes in natural populations. Sequences of Adh were obtained for neo-X and neo-Y chromosomes of D. a. americana, and for Adh of D. a. texana, in which it is autosomal. No significant sequence differentiation is present between the neo-X and neo-Y chromosomes of D. a. americana or the autosomes of D. a. texana. There is a significantly lower level of sequence diversity on the neo-Y chromosome relative to the neo-X in D. a. americana. This reduction in variability on the neo-Y does not appear to have resulted from a selective sweep. Coalescent simulations of the evolutionary transition of an autosome into a Y chromosome indicate there may be a low level of recombination between the neo-X and neo-Y alleles of Adh and that the effective population size of this chromosome may have been reduced below the expected value of 25% of the autosomal effective size, possibly because of the effects of background selection or sexual selection.


Chemical biology--identification of small molecule modulators of cellular activity by natural product inspired synthesis.
The aim of this tutorial review is to introduce the reader to the concept, synthesis and application of natural product-inspired compound collections as an important field in chemical biology. This review will discuss how potentially interesting scaffolds can be identified (structural classification of natural products), synthesized in an appropriate manner (including stereoselective transformations for solid phase-bound compounds) and tested in biological assays (cell-based screening as well as biochemical in vitro assays). These approaches will provide the opportunity to identify new and interesting compounds as well as new targets for chemical biology and medicinal chemistry research.


Preliminary evidence for the conversion of dog renin into a higher-molecular-weight form by cold storage.
1. A soluble fraction of renal cortical homogenate (cytosol) and renin extracted from isolated renin granules of the dog kidney were kept at 0 degrees C. 2. Although the molecular weight of the renin in the cytosol on day 1 was estimated to be about 40 000 by gel filtration, the renin was completely converted into a higher-molecular-weight form (60 000) by day 7. The renin in the granules kept its molecular size of 40 000 during cold storage. 3. This type of molecular-weight conversion could be performed without protease inhibitors. 4. Dithiothreitol neither inhibited the conversion into the higher-molecular-weight form of renin during cold storage nor led to a reduction in the molecular weight of renin, although the oxidation of thiol groups has been proposed as the mechanism for the molecular-weight conversion of renin. 5. Keeping a mixture of renin from the granules and crude renin-binding substance at 0 degrees C for 7 days resulted in the conversion of the renin into the higher-molecular-weight form, indicating that the renin-binding substance we have previously described is required for the conversion during cold storage. 6. Acidification caused the higher-molecular-weight form of renin formed in the cytosol to change to the lower-molecular-weight form, with a concomitant increase in activity of about 50%.


Amycolatopsis sacchari sp. nov., a moderately thermophilic actinomycete isolated from vegetable matter.
The taxonomic position of a group of moderately thermophilic actinomycetes isolated from vegetable matter was determined using a suite of genotypic and phenotypic properties. The organisms were found to share a range of chemical and morphological markers typical of members of the genus Amycolatopsis. A representative of the group, strain K24T, formed a distinct phyletic line within the range of variation occupied by the genus Amycolatopsis in the 16S rDNA tree. The strains have many phenotypic properties in common and some of these distinguish the group from representatives of the validly described species of Amycolatopsis. It is clear from the combined datasets that the strains merit recognition as a new species of Amycolatopsis. The name proposed for the new species is Amycolatopsis sacchari; the type strain is K24T (= DSM 44468T = KCTC 9863T).