Prevalence, patterns, associated factors and severity of substance use among psychotic patients in southern Thailand.
Co-occurring substance use in psychotic patients causes many subsequences including increased illness severity, decreased medication compliance, higher relapse rates, more hospitalizations, and legal problems. We aim to investigate the prevalence, patterns, associated factors and severity of substance use risk among psychotic patients in southern Thailand. Psychotic out-patients were screened with the Alcohol, Smoking, Substance Involvement Screening Test (ASSIST) for their history of substance use in the past three months and categorized as None-to-Low Risk (NLR) or Moderate-to-High Risk (MHR) levels. Multivariate logistic regression was used to examine the associated factors of substance use risk-level. The associations between substance use risk-level and emotional and behavioural symptoms, functional status and family functional status were examined using multivariate linear regression analysis. Of 663 participants screened, 322 (48.6%) used at least one substance in the past three months. Tobacco was the most common substance used (47.2%). The factors associated with a higher risk of any substance use were male gender, young age group, low level of education, being employed and being diagnosed with schizophrenia. A higher number of emotional and behavioural symptoms was significantly associated with higher substance use risk-level. In conclusion, the prevalence of substance use in psychotic patients was high and associated with their emotional and behavioural symptoms. Recommendations for implementation of screening and early intervention programs of substance-related problems in psychotic patients are important for preventing unwanted outcomes.


The Differential Expression of the Inflammasomes in Adipose Tissue and Colon Influences the Development of Colon Cancer in a Context of Obesity by Regulating Intestinal Inflammation.
Inflammasomes maintain tissue homeostasis and their altered regulation in the colon, and the adipose tissue (AT) leads to chronic activation of inflammatory pathways promoting colon cancer (CC) development. We aimed to analyze the potential involvement of inflammasomes in obesity-associated CC. Ninety-nine volunteers [61 with obesity (OB) and 38 normoponderal (NP)] further subclassified according to the approved protocol for the diagnosis of CC (58 without CC and 41 with CC) were included in the case-control study. CC (P<0.01) and obesity (P<0.01) were accompanied by increased mRNA levels of NLRP3, NLRP6, ASC, IL1B and NOD2 in VAT. Contrarily, patients with CC exhibited a downregulation of NLRP6 and IL18 in their colon. Additionally, we revealed that the decreased Nlrp1 (P<0.05), Nlrp3 (P<0.01) and Nlrp6 (P<0.01) mRNA levels in the colon from obese rats significantly increase (P<0.05) after caloric restriction. Adipocyte-conditioned media obtained from subjects with obesity reduced (P<0.01) the mRNA of NLRP3 as well as molecules involved in maintaining the intestinal integrity (MUC2, CLDN1 and TJP1) and the anti-inflammatory factors FGF21, KLF4, and IL33 and in HT-29 cells. We also found that the knockdown of NLRP6 in HT-29 cells significantly upregulated (P<0.05) the mRNA of NLRP1 and NLRP3 and inhibited (P<0.05) the expression levels of MUC2. Finally, we showed that the incubation of HT-29 with Akkermansia muciniphila influence (P<0.05) the inflammasome expression profile as well as intestinal integrity-related genes and aberrant inflammation. These findings provide evidence that the downregulated levels of NLRP6 and IL18 in the colon from patients with CC may be responsible for a reduced intestinal-barrier integrity, triggering local inflammation, which in turn acts on the dysfunctional AT in obesity, increasing the expression of different inflammasome components and flaring up a vicious cycle of uncontrollable inflammatory cascades that favours a pro-tumorigenic microenvironment.


Effect of Endoxylanase and Iron Oxide Nanoparticles on Performance and Histopathological Features in Broilers.
Endoxylanase enzyme is used as poultry feed additive to degrade anti-nutritional factors like non-starch polysaccharides. Moreover, iron is one of the most important trace elements, and its deficiency can lead to various pathological conditions and stunted growth. In the current study, a combination of xylanase and iron nanoparticles is used to assess the overall effect on poultry growth. Endoxylanase enzyme was obtained from Aspergillus awamori SAIB-17 (identified on the basis of internal transcribed spacer (ITS) sequence analysis). The characterization of purified endoxylanase revealed that the Km and Vmax were 0.25 g/ml and 833.33 nkat/ml/s, respectively. Effect of pH and temperature showed that pH 4.5 and temperature 45 °C was best for enzyme activity. Iron nanoparticles were synthesized by co-precipitation of ferric chloride and ferrous chloride. Characterization of nanoparticles using X-ray diffraction (XRD) and SEM revealed that the mean diameter of synthesized iron nanoparticles was around 50 nm. These nanoparticles have no inhibitory effect on endoxylanase up to concentration of 20 ppm. Iron oxide nanoparticles along with endoxylanase were used as additives in different concentrations and were fed to the groups of broiler chicks. It was observed that the group fed with 40 nkat of endoxylanase and 15 ppm of iron nanoparticles showed 54.5% (2010 ± 103.58) more weight gain by the fifth week as compared with the control group. The iron analysis in the muscles showed no increase in iron concentration while histopathology slides showed no morphological changes in liver cells. The combination of iron oxide nanoparticles and xylanase proved to have great potential to be used in poultry feed for large-scale meat production without any toxicological effects.


Accurate stacked-sheet counting method based on deep learning.
The accurate counting of laminated sheets, such as packing or printing sheets in industry, is extremely important because it greatly affects the economic cost. However, the different thicknesses, adhesion properties, and breakage points and the low contrast of sheets remain challenges to traditional counting methods based on image processing. This paper proposes a new stacked-sheet counting method with a deep learning approach using the U-Net architecture. A specific dataset according to the characteristics of stack side images is collected. The stripe of the center line of each sheet is used for semantic segmentation, and the complete side images of the slices are segmented via training with small image patches and testing with original large images. With this model, each pixel is classified by multi-layer convolution and deconvolution to determine whether it is the target object to be detected. After the model is trained, the test set is used to test the model, and a center region segmentation map based on the pixel points is obtained. By calculating the statistical median value of centerline points across different sections in these segmented images, the number of sheets can be obtained. Compared with traditional image algorithms in real product counting experiments, the proposed method can achieve better performance with higher accuracy and a lower error rate.


Keratin 79 identifies a novel population of migratory epithelial cells that initiates hair canal morphogenesis and regeneration.
The formation of epithelial tubes underlies the development of diverse organs. In the skin, hair follicles resemble tube-like structures with lumens that are generated through poorly understood cellular rearrangements. Here, we show that creation of the hair follicle lumen is mediated by early outward movement of keratinocytes from within the cores of developing hair buds. These migratory keratinocytes express keratin 79 (K79) and stream out of the hair germ and into the epidermis prior to lumen formation in the embryo. Remarkably, this process is recapitulated during hair regeneration in the adult mouse, when K79(+) cells migrate out of the reactivated secondary hair germ prior to formation of a new hair canal. During homeostasis, K79(+) cells line the hair follicle infundibulum, a domain we show to be multilayered, biochemically distinct and maintained by Lrig1(+) stem cell-derived progeny. Upward movement of these cells sustains the infundibulum, while perturbation of this domain during acne progression is often accompanied by loss of K79. Our findings uncover previously unappreciated long-distance cell movements throughout the life cycle of the hair follicle, and suggest a novel mechanism by which the follicle generates its hollow core through outward cell migration.


Do tax and subsidy on unhealthy food induce consumer consumption for healthy food? Evidence from experiment in Surabaya, Indonesia.
This study investigates the impact of price changes on decision to buy particular foods among adults in Surabaya, East Java, Indonesia. Tax and subsidy were introduced for influencing decision buy particular foods, especially unhealthy foods that are predicted have consequences on health outcome. Unhealthy foods such as fast foods or junk foods, sugar sweetened beverages (SSB), salty, and fatty foods are reported as one of main causes of obesity incidence in most countries, such as Indonesia. Computer laboratory experimental is designed for investigating the impacts of difference rates of taxes and subsidy on decision to buy foods. Taxes and subsidies rate are designed from low (5%), moderate (15%), and high rates (25%). The findings are as follows. Firstly, participants do not respond immediately to price changes, that is, higher prices due to taxes and lower prices due to subsidies. Economic theory suggests that consumers demand for basic need such as foods is sensitive with change in price, they respond inversely to the price changes. However, the finding in this experimental study do not support this theory prediction. Secondly, 15% and subsidies are the threshold and the larger taxes and rate for changing consumer's choices on targeted foods. This study concludes that low rates of taxes and subsidies for unhealthy and healthy foods make these targeted foods affordable for majority of consumers. Besides introducing higher rates of taxes and subsidies, promoting and encouraging healthy life style such as consuming fresh and healthy foods is another alternative policy option.


An examination of linkage of schizophrenia and schizoaffective disorder to the pseudoautosomal region (Xp22.3)
We investigated linkage between schizophrenia and the loci DXYS14, DXYS17, and MIC2 within the pseudoautosomal region in 85 families with two or more siblings suffering from schizophrenia or schizoaffective disorder. A maximum lod score of 2.44 was reached at MIC2, with a dominant model of inheritance at a recombination fraction of 0.367 in females and 0.046 in males (a F:M sex ratio > 1, i.e. opposite to that expected with a pseudoautosomal locus). Evidence consistent with linkage (P = 0.01) was also obtained with a sibling pair analysis at the MIC2 locus. These data do not support (although they do not definitively exclude) a locus within the pseudoautosomal region; they are consistent with the presence of a gene that predisposes to schizophrenia in the sex-specific regions of the X and Y chromosomes.


Relaxation dispersion NMR spectroscopy as a tool for detailed studies of protein folding.
Characterization of the mechanisms by which proteins fold into their native conformations is important not only for protein structure prediction and design but also because protein misfolding intermediates may play critical roles in fibril formation that are commonplace in neurodegenerative disorders. In practice, the study of folding pathways is complicated by the fact that for the most part intermediates are low-populated and short-lived so that biophysical studies are difficult. Due to recent methodological advances, relaxation dispersion NMR spectroscopy has emerged as a particularly powerful tool to obtain high-resolution structural information about protein folding events on the millisecond timescale. Applications of the methodology to study the folding of SH3 domains have shown that folding proceeds via previously undetected on-pathway intermediates, sometimes stabilized by nonnative long-range interactions. The relaxation dispersion approach provides a detailed kinetic and thermodynamic description of the folding process as well as the promise of obtaining an atomic level structural description of intermediate states. We review the concerted application of a variety of recently developed NMR relaxation dispersion experiments to obtain a "high-resolution" picture of the folding pathway of the A39V/N53P/V55L Fyn SH3 domain.


Ultrasound guided axillary brachial plexus plexus block. Part 2 - technical issues.
Axillary brachial plexus block is one of the most frequently employed peripheral blocks. The popularity of axillary block stems from its success as a safe and relatively easy technique with numerous applications. The technique of axillary block has evolved. It was modified after the development of precise nerve localization modalities. Currently, ultrasound is the most important localization technique for regional anaesthesia. Ultrasound-guided axillary block encompasses a spectrum of techniques. The selection of a specific technique can be adjusted to an operator's individual level of skill and proficiency. Axillary block under US-guidance can be performed using a traditional perivascular method and by placing a selective blockade of individual nerves that supply the surgical area. Regardless of the selected method, it enables the incorporation of individual patient anatomical variation in an anaesthesia plan. This paper discusses the technical details and efficacy issues of US-guided axillary brachial plexus block techniques.


C6-ceramide enhances Interleukin-12-mediated T helper type 1 cell responses through a cyclooxygenase-2-dependent pathway.
Ceramides, lipid molecules located predominantly within the plasma membrane of a cell, can function as second messengers, and have been known to carry out a number of cellular functions. T helper type 1 (Th1) immune responses are known to be involved in the cellular immunity, which is crucial in the cancer and allergy immunotherapy. This study was designed to evaluate the effects of ceramides on T helper cell responses and their underlying mechanisms. We demonstrated that a cell-permeable C6-ceramide (C6) together with IL-12 enhanced Th1 cell differentiation, whereas C6 alone had no effects, as demonstrated by the increased populations of IFN-γ expressing CD4(+) T cells and the up-regulation of IFN-γ production from CD4(+) T cells. In contrast, C2-ceramide and long chain ceramides (C16 and C24) did not affect the Th1 responses. C6 treatment was shown to increase the expression of T-bet, a master transcription factor of Th1 responses, in a dose-dependent fashion. Furthermore, C6 increased the expression of cyclooxygenase-2 (COX-2) in CD4(+) T cells. The C6-mediated increase of IFN-γ production and IFN-γ expressing CD4(+) T cell populations were significantly suppressed by a COX-2 specific inhibitor (NS-398) in a dose-dependent manner. T-bet expression was also decreased by NS-398 treatment, thereby indicating that C6 ceramide enhances Th1 responses via a COX-2 dependent pathway. This result demonstrates that C6 may be utilized in therapies for the treatment of immune diseases such cancer and allergy by enhancing the Th1 activity.


Incidence of Attention Deficit Hyperactivity Disorder (ADHD) Diagnoses in Navarre (Spain) from 2003 to 2019.
(1) Background: Attention deficit hyperactivity disorder (ADHD) constitutes one of the leading mental health and behavioral disorders in childhood and adolescence. The main objective of this study was to analyze the time trend in the incidence of ADHD diagnoses in Navarre (Spain) from 2003 to 2019 in children and adolescents from 5 to 19 years old. Additionally, the seasonal trends of ADHD incidence and ADHD prevalence were determined. (2) Methods: A population-based observational retrospective study, which included people born between 1991 and 2011 and who attended compulsory education between 2007 and 2017 in Navarre (Spain), was carried out with data from both the Education and Health Department databases. (3) Results: The incidence rate increased from 4.18 cases per 1000 person-years in 2003 to 7.43 cases per 1000 person-years in 2009, before decreasing progressively to 2.1 cases per 1000 person-years in 2019. A peak incidence rate at 7-8 years of age was observed, which is consistent across the study period and for both genders. Males were more than twice as likely to be diagnosed with ADHD than females, with similar time trends in both. A seasonal pattern in ADHD diagnosis was found, with peaks in February-March and the lowest rates in the summer months. Inattentive cases were much more frequent than hyperactive cases, whereas combined cases remained low across the study period. (4) Conclusions: In this age-period-cohort analysis, a clear period and age effect was observed. We found a decreasing trend in the ADHD incidence rate since 2015. Further research is needed to confirm whether a change of trend is occurring globally.


Two thumbs up: using popular films in introductory aging courses.
Good teaching requires thoughtful planning and creative thinking, especially when trying to engage students in material that is unfamiliar to them or encumbered by stereotypes, like aging. Classic and contemporary media can provide unique teaching opportunities in gerontology classrooms. Popular films can have a powerful influence over viewers' attitudes and perceptions, and spur in-depth discussions of aging-related topics common to introductory aging courses (e.g., ageism, abuse, inequality, caregiving, healthy aging, and intimate relationships). Additionally, films appeal to multiple learning styles, engaging a variety of learners. This article examines the value of using films in introductory aging courses, offers strategies for incorporating films in the gerontology classroom, suggests sample activities and assignments that pair popular films with aging course topics, identifies challenges of using film in various classrooms settings, and provides a detailed typology of films on each of the following aging topics: ageism and stereotypes, cognitive impairment, death and dying, diversity, family relationships, health and wellness, sexuality and intimacy, and work and retirement.


Relating Divisive Normalization to Neuronal Response Variability.
Cortical responses to repeated presentations of a sensory stimulus are variable. This variability is sensitive to several stimulus dimensions, suggesting that it may carry useful information beyond the average firing rate. Many experimental manipulations that affect response variability are also known to engage divisive normalization, a widespread operation that describes neuronal activity as the ratio of a numerator (representing the excitatory stimulus drive) and denominator (the normalization signal). Although it has been suggested that normalization affects response variability, we lack a quantitative framework to determine the relation between the two. Here we extend the standard normalization model, by treating the numerator and the normalization signal as variable quantities. The resulting model predicts a general stabilizing effect of normalization on neuronal responses, and allows us to infer the single-trial normalization strength, a quantity that cannot be measured directly. We test the model on neuronal responses to stimuli of varying contrast, recorded in primary visual cortex of male macaques. We find that neurons that are more strongly normalized fire more reliably, and response variability and pairwise noise correlations are reduced during trials in which normalization is inferred to be strong. Our results thus suggest a novel functional role for normalization, namely, modulating response variability. Our framework could enable a direct quantification of the impact of single-trial normalization strength on the accuracy of perceptual judgments, and can be readily applied to other sensory and nonsensory factors.SIGNIFICANCE STATEMENT Divisive normalization is a widespread neural operation across sensory and nonsensory brain areas, which describes neuronal responses as the ratio between the excitatory drive to the neuron and a normalization signal. Normalization plays a key role in several important computations, including adjusting the neuron's dynamic range, reducing redundancy, and facilitating probabilistic inference. However, the relation between normalization and neuronal response variability (a fundamental aspect of neural coding) remains unclear. Here we develop a new model and test it on primary visual cortex responses. We show that normalization has a stabilizing effect on neuronal activity, beyond the known suppression of firing rate. This modulation of variability suggests a new functional role for normalization in neural coding and perception.


Hybridization-based in situ sequencing (HybISS) for spatially resolved transcriptomics in human and mouse brain tissue.
Visualization of the transcriptome in situ has proven to be a valuable tool in exploring single-cell RNA-sequencing data, providing an additional spatial dimension to investigate multiplexed gene expression, cell types, disease architecture or even data driven discoveries. In situ sequencing (ISS) method based on padlock probes and rolling circle amplification has been used to spatially resolve gene transcripts in tissue sections of various origins. Here, we describe the next iteration of ISS, HybISS, hybridization-based in situ sequencing. Modifications in probe design allows for a new barcoding system via sequence-by-hybridization chemistry for improved spatial detection of RNA transcripts. Due to the amplification of probes, amplicons can be visualized with standard epifluorescence microscopes for high-throughput efficiency and the new sequencing chemistry removes limitations bound by sequence-by-ligation chemistry of ISS. HybISS design allows for increased flexibility and multiplexing, increased signal-to-noise, all without compromising throughput efficiency of imaging large fields of view. Moreover, the current protocol is demonstrated to work on human brain tissue samples, a source that has proven to be difficult to work with image-based spatial analysis techniques. Overall, HybISS technology works as a targeted amplification detection method for improved spatial transcriptomic visualization, and importantly, with an ease of implementation.


Ammonium hydroxide hydrolysis: a valuable support in the MALDI-TOF mass spectrometry analysis of Lipid A fatty acid distribution.
Lipid A is the lipophilic moiety of lipopolysaccharides (LPSs), the major components of the external membrane of almost all gram-negative bacteria. It is responsible for the toxicity of LPS and has a heterogeneous structure composed of a bis-phosphorylated glucosamine disaccharide backbone that is acylated at the positions 2, 3 of the GlcN I (proximal) and GlcN II (distal) residue with O- and N-linked 3-hydroxy fatty acids (primary substitution). These fatty acids are further acylated by means of their 3-hydroxy groups (secondary substitution). The toxicity of Lipid A is dependent on its primary structure; the number, the length, and the distribution of the fatty acids on the disaccharide backbone strongly influence the endotoxic activity. In this paper a general and easy methodology to obtain secondary fatty acid distribution, which is one of the most difficult issues in the structural determination of Lipid A, is proposed. The method combines ammonium hydroxide hydrolysis and matrix assisted laser desorption ionization (MALDI)-mass spectrometry analysis and has been successfully proven with five different Lipid A species. The procedure exploits the lower stability under mild alkaline conditions of acyl and acyloxyacyl esters with respect to that of the acyl and acyloxyacyl amides. The partially degraded Lipid A species obtained are analyzed by MALDI-MS. The generality of this approach was tested on five Lipid As, namely those arising from Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca, Pseudomonas reactans, and Burkholderia caryophylli.


Microstructural mechanisms of hysteresis and transformation width in NiTi alloy from molecular dynamics simulations.
Martensitic transformations in shape memory alloys are often accompanied by thermal hysteresis, and engineering this property is of prime scientific interest. The martensitic transformation can be characterized as thermoelastic, where the extent of the transformation is determined by a balance between thermodynamic driving force and stored elastic energy. Here we used molecular dynamics simulations of the NiTi alloy to explore hysteresis-inducing mechanisms and thermoelastic behavior by progressively increasing microstructural constraints from single crystals to bi-crystals to polycrystals. In defect-free single crystals, the austenite-martensite interface moves unimpeded with a high velocity. In bi-crystals, grain boundaries act as significant obstacles to the transformation and produce hysteresis by requiring additional nucleation events. In polycrystals, the transformation is further limited by the thermoelastic balance. The stored elastic energy can be converted to mechanisms of non-elastic strain accommodation, which also produce hysteresis. We further demonstrated that the thermoelastic behavior can be controlled by adjusting microstructural constraints.


Internal cranial expansion surgery for the treatment of refractory idiopathic intracranial hypertension.
Idiopathic intracranial hypertension (IIH) may be refractory to available medical and surgical therapies. Patients with this condition may suffer from intractable headaches, experience visual deterioration, or have other symptoms related to elevated intracranial pressure. Internal cranial expansion (ICE) is a novel surgical procedure that the authors have developed for the treatment of patients with this condition. Here, they describe ICE and present their initial experience in using this surgical procedure for the treatment of patients with refractory IIH. The authors conducted a retrospective review of 10 consecutive patients who underwent ICE for the treatment of IIH during a 5-year period. Preoperative and postoperative clinical parameters including patient symptoms, presence of papilledema, and available ICP or CSF opening pressures were compared. Procedural details and complications were noted. Intracranial volume increases were calculated using available pre- and postoperative CT scans. Follow-up for the 10 patients in this series ranged from 1 to 39.6 months (mean 15.5 months). Technically successful ICE was performed in all patients within the cohort. Surgical complications included a single postoperative seizure in one patient and a sagittal sinus tear with no clinical sequelae in another patient. At the time of last follow-up, 7 (70%) of 10 patients were either symptomatically improved or asymptomatic. Six (67%) of 9 patients with preoperative headaches had reduction or resolution of this symptom, and all patients (4 of 4) with preoperative papilledema had a reduction in or complete resolution of this sign. Postoperative ICP or CSF opening pressures were normal in all patients (4 of 4) tested. Postoperative intracranial volume expansion ranged between 3.8% and 12%. Internal cranial expansion is a safe and effective surgery for the treatment of patients with refractory IIH. This surgery expands the intracranial volume and thus promotes ICP normalization, which may lead to the reduction or complete resolution of the signs and symptoms of IIH. Internal cranial expansion may be used as part of a multidisciplinary management approach in the treatment of refractory IIH.


BT cationic peptides: small peptides that modulate innate immune responses of chicken heterophils and monocytes.
Neonatal poultry exhibit a transient susceptibility to infectious diseases during the first week of life that stems from inefficient host defense mechanisms. Yet, the initial host immune response to pathogens is a critical determinant of disease resistance and susceptibility. With this context in mind, novel ways to stimulate or modulate the hosts' natural immune response is emerging as an important area of interest for food animal producers including the poultry industry. Specifically, we have been investigating new modulation strategies tailored around the selective stimulation of the host's immune system, and particularly rapid acting innate immunity, as an alternative to direct targeting of microbial pathogens. One such approach that we have been investigating is the use of a group of cationic peptides produced by a Gram-positive soil bacterium, Brevibacillus texasporus (BT peptides). We have previously shown that, provided as a feed additive, BT peptides significantly induced a concentration-dependent protection against cecal colonization and extraintestinal colonization by Salmonella enterica serovar Enteritidis (SE). This protection is not the result of direct antibacterial activity of the BT peptides on the SE since the concentrations used were below the minimum inhibitory concentration for SE. We also found that BT are not absorbed in the intestine, but still induce a significant up-regulation in the functional efficiency of peripheral blood heterophils and monocytes. The mechanisms of this immune modulation are unknown. Here, using in vitro models for measuring: (1) leukocyte oxidative burst, (2) changes in leukocyte cytokine and chemokines gene expression profiles, and (3) phosphorylation of the mitogen activated protein kinases (MAPKs) in leukocytes, we evaluated the role of BT peptides as priming mediators for heterophil and monocyte responses at the level of cell function, gene transcription/expression, and cell phosphorylation following stimulation with inflammatory agonists. BT peptides primed both heterophils and monocytes for an increased oxidative burst and up-regulation in transcription of the pro-inflammatory cytokines IL-1β and IL-6 and inflammatory chemokines CXCLi1 and CXCLi2 induced by inflammatory agonists. In addition, BT peptides induced a rapid (10min) phosphorylation and activation of the extracellular signal-regulated kinase (ERK1/2) and p38 kinase pathways in primary chicken heterophils. Taken together, we conclude that BT peptides, acting through MAPK pathways, enhance leukocyte functional and pro-inflammatory cytokine and chemokine gene transcription activities. These small cationic peptides may prove useful as immune modulators in neonatal poultry.


Mechanisms that regulate Epstein-Barr virus EBNA-1 gene transcription during restricted latency are conserved among lymphocryptoviruses of Old World primates.
Epstein-Barr virus (EBV), the only known human lymphocryptovirus (LCV), displays a remarkable degree of genetic and biologic identity to LCVs that infect Old World primates. Within their natural hosts, infection by these viruses recapitulates many key aspects of EBV infection, including the establishment of long-term latency within B lymphocytes, and is therefore a potentially valuable animal model of EBV infection. However, it is unclear whether these LCVs have adopted or maintained the same mechanisms used by EBV to express essential viral proteins, such as EBNA-1, in the face of cell-mediated repression of EBV gene expression that occurs upon establishment of the asymptomatic carrier state. To address this issue, we determined whether the endogenous LCVs of baboon (Cercopithecine herpesvirus 12) and rhesus macaque (Cercopithecine herpesvirus 15) have the functional equivalent of the EBV promoter Qp, which mediates exclusive expression of EBNA-1 during the restricted programs of EBV latency associated with the carrier state. Our results indicate that (i) both the baboon and rhesus macaque LCVs have a genomic locus that is highly homologous to the EBV Qp region, (ii) key cis-regulatory elements of Qp are conserved in these LCV genomes and compose promoters that are functionally indistinguishable from EBV Qp, and (iii) EBNA-1 transcripts identical in structure to EBV Qp-specific EBNA-1 mRNAs are present in nonhuman LCV-infected cells, demonstrating that these Qp homologs are indeed utilized as alternative EBNA-1 promoters. These observations indicate that the molecular mechanisms which regulate EBV gene expression during restricted latency have been conserved among the LCVs. The contribution of these mechanisms to viral persistence in vivo can now be experimentally tested in nonhuman primate models of LCV infection.


Occupational Safety and Health Among Young Workers in the Nordic Countries: A Systematic Literature Review.
This review aimed to identify risk factors for occupational accidents and illnesses among young workers in the Nordic countries and to attain knowledge on specific vulnerable groups within the young working force that may need special attention. We conducted a systematic review from 1994 to 2014 using five online databases. Of the 12,528 retrieved articles, 54 met the review criteria and were quality assessed, in which data were extracted focusing on identifying occupational safety, health risk factors, and vulnerable groups among the young workers. The review shows that mechanical factors such as heavy lifting, psychosocial factors such as low control over work pace, and organizational factors such as safety climate are all associated with increased injury risk for young Nordic workers. Results show that exposures to chemical substances were associated with skin reactions, e.g., hand eczema. Heavy lifting and awkward postures were risk factors for low back pain, and high job demands were risk factors for mental health outcomes. The review identified young unskilled workers including school drop-out workers as particularly vulnerable groups when it comes to occupational accidents. In addition, apprentices and young skilled workers were found to be vulnerable to work-related illnesses. It is essential to avoid stereotyping young Nordic workers into one group using only age as a factor, as young workers are a heterogeneous group and their vulnerabilities to occupational safety and health risks are contextual. Politicians, researchers, and practitioners should account for this complexity in the education, training and organization of work, and workplace health and safety culture.


Predicting COVID-19 Vaccination Intention: The Roles of Institutional Trust, Perceived Vaccine Safety, and Interdependent Self-Construal.
The purpose of this study is to explore the process through which institutional trust influences individuals' vaccination intentions for a coronavirus vaccine. We collected data from a national representative sample of the Spanish population (N = 500) from an online research panel to test the proposed conceptual model. Results revealed a positive association of institutional trust with coronavirus vaccination intentions. This effect was mediated by perceived vaccine safety and positive attitude toward the vaccine. Furthermore, findings corroborate the moderating effect of interdependent self-construal on the indirect relationship between institutional trust and vaccination intention, implying that this effect was stronger for individuals with higher levels of interdependent orientation. Findings have implications for the development of evidence-based health communication strategies for encouraging COVID-19 vaccination.


Comparative effectiveness of homoeopathic vs. conventional therapy in usual care of atopic eczema in children: long-term medical and economic outcomes.
One in five children visiting a homeopathic physician suffers from atopic eczema. We aimed to examine the long-term effectiveness, safety and costs of homoeopathic vs. conventional treatment in usual medical care of children with atopic eczema. In this prospective multi-centre comparative observational non-randomized rater-blinded study, 135 children (48 homoeopathy, 87 conventional) with mild to moderate atopic eczema were included by their respective physicians. Depending on the specialisation of the physician, the primary treatment was either standard conventional treatment or individualized homeopathy as delivered in routine medical care. The main outcome was the SCORAD (SCORing Atopic Dermatitis) at 36 months by a blinded rater. Further outcomes included quality of life, conventional medicine consumption, safety and disease related costs at six, 12 and 36 months after baseline. A multilevel ANCOVA was used, with physician as random effect and the following fixed effects: age, gender, baseline value, severity score, social class and parents' expectation. The adjusted mean SCORAD showed no significant differences between the groups at 36 months (13.7 95% CI [7.9-19.5] vs. 14.9 [10.4-19.4], p = 0.741). The SCORAD response rates at 36 months were similar in both groups (33% response: homoeopathic 63.9% vs. conventional 64.5%, p = 0.94; 50% response: 52.0% vs. 52.3%, p = 0.974). Total costs were higher in the homoeopathic versus the conventional group (months 31-36 200.54 Euro [132.33-268.76] vs. 68.86 Euro [9.13-128.58], p = 0.005). Taking patient preferences into account, while being unable to rule out residual confounding, in this long-term observational study, the effects of homoeopathic treatment were not superior to conventional treatment for children with mild to moderate atopic eczema, but involved higher costs.


Hypocotyl expression and light downregulation of the soybean tubulin gene, tubB1.
The tubB1 beta-tubulin gene of Glycine max (previously named s beta 1) is highly expressed only in rapidly elongating regions of etiolated seedling hypocotyls and this expression is strongly downregulated when the seedlings are exposed to light. Primer extension demonstrated that the gene was transcribed in these tissues and contained two sites of transcriptional initiation. To determine the mechanism regulating tubB1 expression, a chimeric reporter gene was constructed by fusing 5' upstream regions of tubB1 to a promoterless beta-glucuronidase (GUS) gene and these constructs were introduced into protoplasts by electroporation. Strong transient expression of the reporter gene was obtained after electroporation of chimeric constructs containing 1 kb of tubB1 5' upstream sequence into tobacco protoplasts. Deletion of the distal most 300 bp from the 5' sequence of tubB1 enhanced expression, suggesting the possibility of a negative transcriptional regulator in this region. Additional deletions of the 5' sequence reduced expression substantially. Constructs containing a tubB1 3' terminus were expressed at much lower levels than those containing a nopaline synthase (NOS) 3' terminus. The tubB1-GUS chimeric gene also was introduced into tobacco by Agrobacterium-mediated Ti plasmid transformation and the organ-specific expression pattern of the chimeric gene was determined in seedlings of the transgenic plants. Hypocotyls exhibited strong GUS activity when the seedlings were germinated in darkness, but lacked the GUS enzyme when the seedlings were germinated in the light.(ABSTRACT TRUNCATED AT 250 WORDS)


The WHO maternal near miss approach: consequences at Malawian District level.
WHO proposes a set of organ-failure based criteria for maternal near miss. Our objective was to evaluate what implementation of these criteria would mean for the analysis of a cohort of 386 women in Thyolo District, Malawi, who sustained severe acute maternal morbidity according to disease-based criteria. A WHO Maternal Near Miss (MNM) Tool, created to compare disease-, intervention- and organ-failure based criteria for maternal near miss, was completed for each woman, based on a review of all available medical records. Using disease-based criteria developed for the local setting, 341 (88%) of the 386 women fulfilled the WHO disease-based criteria provided by the WHO MNM Tool, 179 (46%) fulfilled the intervention-based criteria, and only 85 (22%) the suggested organ-failure based criteria. In this low-resource setting, application of these organ-failure based criteria that require relatively sophisticated laboratory and clinical monitoring underestimates the occurrence of maternal near miss. Therefore, these criteria and the suggested WHO approach may not be suited to compare maternal near miss across all settings.


[A new material in neurosurgery].
A new alcoholic prolamine solution is now available (ETHIBLOC, supplied by Ethicon, Norderstedt) which can be used in neurosurgery for a variety of purposes. This material hardens quickly in an aqueous medium. Animal experiments have not shown any damaging action on the brain, meninges, or bone. The excellent tolerance of the material was seen in a total of 22 patients operated on with the help of Ethibloc. It is particularly well suited for closing frontal CSF fistulas, incised frontal sinuses, and for rearranging or shunting in case of cerebral vascular deformities. It can also be used as a carrier of antibiotics in osteomyelitis, for covering up minor defects of the dura mater, for dura sealing, and for closing the bone after trephination. The preparation is transformed completely to connective tissue within a period of 40 to 90 days. New techniques of surgery are described which have become possible by the use of Ethibloc.


Interaction of Mycoplasma dispar and Mycoplasma agalactiae subsp. bovis with bovine alveolar macrophages and bovine lacteal polymorphonuclear leukocytes.
Mycoplasma dispar and Mycoplasma agalactiae subsp. bovis survived or grew in cultures of bovine lacteal polymorphonuclear leukocytes or bovine alveolar macrophages. In the presence of specific bovine antibody, macrophages and polymorphonuclear leukocytes appeared to kill both species of mycoplasma. Specific rabbit antisera also promoted the killing of these mycoplasmas by bovine macrophages but had no demonstrable activity for bovine polymorphonuclear leukocytes. It is suggested that phagocytosis of these mycoplasmas by bovine cells occurs only in the presence of specific antibody. The experiments also indicate that differences exist between bovine polymorphonuclear leukocytes and macrophages with regard to their receptor sites for immunoglobulins.


Analysis of cortisol secretion in hormonally inactive adrenocortical incidentalomas: study of in vitro steroid secretion and immunohistochemical localization of steroidogenic enzymes.
Adrenal incidentalomas have recently increased in incidence, and thus it has become important to establish clinical management of these patients. It is also important to evaluate whether these tumors are different from preclinical or overt Cushing's syndrome in their steroidogenesis. In this study, we therefore examined steroidogenesis of hormonally inactive adrenal incidentalomas via short-term culture of tumor specimens, in addition to an immunohistochemical study of steroidogenic enzymes. Five patients (two men and three women) diagnosed with adrenocortical incidentaloma without any clinical signs of adrenocortical hormonal excess except for hypertension and disturbed glucose tolerance, were recruited for this study. Hormonal findings, including circadian rhythms for cortisol and ACTH secretion, the response of ACTH to CRH infusion and results of dexamethasone suppression test were all within normal limits in these patients. Immunoreactivity for all steroidogenic enzymes involved in cortisol production was detected in tumor cells in all cases examined. Results of in vitro steroidogenesis analysis using short-term culture revealed that levels of cortisol secretion varied among the cases. There were no differences in the immunolocalization of steroidogenic enzymes and/or the levels of cortisol secretion between these hormonally inactive tumors and preclinical and/or overt Cushing's syndrome. Dehydroepiandrosterone-sulfotransferase (DHEA-ST) immunoreactivity in nonneoplastic regions was suppressed in one case in which the tumor secreted cortisol similar to preclinical and/or overt Cushing's syndrome. These results demonstrate that the levels of in vitro steroid production and/or the immunolocalization of steroidogenic enzymes in hormonally inactive adrenocortical tumors vary markedly and are not overtly different from those of preclinical and/or overt Cushing's syndrome.


High prevalence of anti-HGV/E2 antibodies in HCV-positive patients with non Hodgkin's lymphoma.
We evaluated in a series of 33 HCV positive (both RT-PCR and HCV RIBA 2 assays) B cell non-Hodgkin's lymphomas (NHL) patients the prevalence of active and inactive HGV infection by HGV RNA assays (RT-PCR) and anti HGV antibodies directed against E2 structural protein (immunoenzimatic method), a reliable serologic marker of past HGV infection followed by viral clearance. We found only one patient with HGV positivity at RT-PCR (3%). Twenty-six of 33 patients were positive for anti HGV/E2 antibodies (78.8%) suggesting past infection. If confirmed, our preliminary data seem to suggest a higher incidence of HGV past infection in our group of HCV positive patients with B cell NHL.


Probiomimetics-Novel Lactobacillus-Mimicking Microparticles Show Anti-Inflammatory and Barrier-Protecting Effects in Gastrointestinal Models.
There is a lack of efficient therapies to treat increasingly prevalent autoimmune diseases, such as inflammatory bowel disease and celiac disease. Membrane vesicles (MVs) isolated from probiotic bacteria have shown tremendous potential for treating intestinal inflammatory diseases. However, possible dilution effects and rapid elimination in the gastrointestinal tract may impair their application. A cell-free and anti-inflammatory therapeutic system-probiomimetics-based on MVs of probiotic bacteria (Lactobacillus casei and Lactobacillus plantarum) coupled to the surface of microparticles is developed. The MVs are isolated and characterized for size and protein content. MV morphology is determined using cryoelectron microscopy and is reported for the first time in this study. MVs are nontoxic against macrophage-like dTHP-1 and enterocyte-like Caco-2 cell lines. Subsequently, the MVs are coupled onto the surface of microparticles according to facile aldehyde-group functionalization to obtain probiomimetics. A significant reduction in proinflammatory TNF-α level (by 86%) is observed with probiomimetics but not with native MVs. Moreover, it is demonstrated that probiomimetics have the ability to ameliorate inflammation-induced loss of intestinal barrier function, indicating their potential for further development into an anti-inflammatory formulation. These engineered simple probiomimetics that elicit striking anti-inflammatory effects are a key step toward therapeutic MV translation.


[Proteolytic activity in rat liver mitochondria].
An enzymatic fraction, possessing the proteolytic activity, was isolated from destructed rat liver mitochondria. The preparation obtained exhibited the distinct activity with membrane proteins and mitochondrial structural proteins used as substrates and dissolved in solutions, containing EDTA and sodium dodecylsulphate, respectively. The proteolytic activity was shown to be due to protease, found in the preparation of mitochondria; it did not depend on the contamination by lysosomal cathepsins.


Adoptive cell therapy using in vitro generated human CD4+ CD25+ regulatory t cells with indirect allospecificity to promote donor-specific transplantation tolerance.
The holy grail in clinical transplantation is the establishment of long-term donor-specific transplantation tolerance with the minimum use of immunosuppressive agents. CD4+ CD25+ regulatory T cells (Tregs) play a crucial role in the prevention of autoimmunity, and appear to mediate transplantation tolerance. Harnessing Tregs for potential adoptive cell therapy to promote donor-specific transplantation tolerance is promising. Here we show that human CD4+ CD25+ Tregs with indirect allospecificity for an HLA-A2 (103-120) peptide can be generated from purified peripheral blood CD4+ CD25+ by priming with HLA-DR0101+ A2- autologous dendritic cells (DCs) pulsed with the A2 peptide in vitro. The antigen specificity for the A2 peptide was demonstrated in functional assays and flow cytometric analysis using a fluorescent tetramer composed of HLA-DR0101 and the A2 peptide. The CD4+ CD25+ Tregs with indirect allospecificity for the A2 peptide showed potent suppression of an indirect alloresponse by effector CD4+ CD25- T-helper cells. Importantly, the selected CD4+ CD25+ Tregs can be expanded substantially to meet a therapeutic end after T-cell receptor (TCR) stimulation by CD3/CD28 antibody-coated beads in the presence of high doses of interleukin-2 (IL-2). The expanded CD4+ CD25+ Tregs highly expressed Foxp3, and retained their suppressive properties and maintained expression of lymphoid homing receptor CD62L. Taken together, these data pave the way for clinical studies using in vitro generated and expanded human CD4+ CD25+ Tregs with indirect allospecificity as therapeutic reagents to promote donor-specific transplantation tolerance.


Clostridium difficile 30 years on: what has, or has not, changed and why?
The report of clindamycin-associated colitis in 1974 by Tedesco et al. [Ann Intern Med 81: 429-33] stimulated an intense search for the cause of this severe complication of antibiotic use. The search culminated in early 1978 in the publication of a series of papers within 3 months that identified the causative agent as Clostridium difficile and its accompanying toxins. Thirty years later we are in the midst of a resurgence of C. difficile infection (CDI) in North America and Europe that is greater than ever previously reported and for which morbidity and mortality appear to be higher than ever seen in the past. The purpose of this review is to highlight the discoveries of the past 30 years that, in my view, have brought us to our current level of understanding of the pathogenesis, prevention and treatment of CDI, and to suggest why a disease thought to be managed so well 30 years ago could now be causing more morbidity and mortality than ever before. In the 21st century the focus should be on better understanding the relationship between the C. difficile organism and the host at the mucosal level, so that biotherapeutic and vaccine strategies for the prevention of CDI can be developed.


Integrated delivery systems. Evolving oligopolies.
The proliferation of Integrated Delivery Systems (IDSs) in regional health care markets has resulted in the movement of these markets from a monopolistic competitive model of behavior to an oligopoly. An oligopoly is synonymous with competition among the few, as a small number of firms supply a dominant share of an industry's total output. The basic characteristics of a market with competition among the few are: (1) A mutual interdependence among the actions and behaviors of competing firms; (2) competition tends to rely on the differentiation of products; (3) significant barriers to entering the market exist; (4) the demand curve for services may be kinked; and (5) firms can benefit from economies of scale. An understanding of these characteristics is essential to the survival of IDSs as regional managed care markets mature.


Noncontact, high-resolution ultrasonic imaging of wood samples using coded chirp waveforms.
A noncontact ultrasonic inspection technique has been developed to study the properties of wood samples in air. The system makes use of two broad bandwidth capacitive transducers, combined with signal processing techniques. A coded chirp signal was used in the current application to provide a waveform that could be postprocessed to provide sufficient sensitivity for transmission across samples of wood. It will be shown in this paper that the signal-to-noise ratio (SNR) can be greatly improved using two signal recovery techniques, namely pulse compression and swept frequency multiplication (SFM). A simulation of both techniques is presented and compared to experimental data. As will be seen from the experimental results, it is possible to perform noncontact ultrasonic experiments to extract a range of useful information such as ring density and the presence of microcracks.


Secondary organic aerosol formation and organic nitrate yield from NO3 oxidation of biogenic hydrocarbons.
The secondary organic aerosol (SOA) mass yields from NO3 oxidation of a series of biogenic volatile organic compounds (BVOCs), consisting of five monoterpenes and one sesquiterpene (α-pinene, β-pinene, Δ-3-carene, limonene, sabinene, and β-caryophyllene), were investigated in a series of continuous flow experiments in a 10 m(3) indoor Teflon chamber. By making in situ measurements of the nitrate radical and employing a kinetics box model, we generate time-dependent yield curves as a function of reacted BVOC. SOA yields varied dramatically among the different BVOCs, from zero for α-pinene to 38-65% for Δ-3-carene and 86% for β-caryophyllene at mass loading of 10 μg m(-3), suggesting that model mechanisms that treat all NO3 + monoterpene reactions equally will lead to errors in predicted SOA depending on each location's mix of BVOC emissions. In most cases, organonitrate is a dominant component of the aerosol produced, but in the case of α-pinene, little organonitrate and no aerosol is formed.


OCLN gene variants identified in three patients with severe neurodevelopmental disorder associated with epilepsy, intellectual disability and malformation of cortical development.
Homozygous OCLN variants have been reported in patients with band-like calcification with simplified gyration and polymicrogyria (BLC-PMG) which is characterized by microcephaly, intracranial calcification and severe developmental delay. The OCLN gene encodes the integral membrane protein, occludin. Herein, we report three additional cases with homozygous OCLN variants that were identified via Trio-WES in two consanguineous unrelated families. We detected a previously reported frameshift variant in two cases in Family 1 and a novel missense variant in a case in Family 2. The potential pathogenicity of both variants in the index cases was investigated using in silico tools, and both variants were determined to be rare and predicted to be pathogenic. All of the presented cases exhibited clinical features in common with earlier reported patients, such as severe intellectual disability, microcephaly, polymicrogyria, epilepsy, hypotonia and severe developmental delay. On the other hand, in addition to the common phenotypic features, our two cases in Family 1 showed features similar to those previously reported in cases from two Turkish families carrying the same frameshift variant, such as renal failure. We herein expand the spectrum of OCLN gene variants with a description of an additional novel homozygous missense variant. The frameshift variant in Turkish cases may thus be a phenotype associated with renal failure in addition to the core phenotype associated with other OCLN gene variants, and such variants could be important for rapid molecular diagnosis and treatment. The frameshift variant in Turkish cases might also be investigated for both a potential founder effect and mutational hot spot.


Open versus endoscopic bone resection of the dorsolateral calcaneal edge: a cadaveric analysis comparing three dimensional CT scans.
It has been claimed that endoscopic calcaneoplasty offers some advantages over open techniques in the surgical treatment of Haglund's deformity due to reduced postoperative complications like stiffness and pain. Bony over-resection places patients at risk of these complications. The resulting question with regard to the quantitative differences of the extent of the bone removed using these two techniques has not yet been answered. The purpose of the study was to determine the resection volume of calcaneal bone for open and endoscopic surgical techniques. 16 feet obtained from body donors were operated on in equal parts using either open surgical or endoscopic techniques, with the technique selected on a random basis. High-resolution CT scans were obtained before and after the interventional procedure and analysed to obtain 3-D polygon models. Post-operative models were subtracted from pre-operative models to provide the volume change resulting from the intervention. This was then correlated with the bone mineral density (BMD) of the preparation. The extent of bony resection was greater in open surgical techniques than in endoscopic approaches. The average volume of bone resection was 0.80 (±0.34) cm(3) in the endoscopic group and 3.04 (±2.91) cm(3) in the group that underwent open surgery. After adjustment for bone mineral density the extent of the resection was significantly larger (p = 0.018) in the group undergoing open surgery. The two groups did not differ significantly with regard to BMD (p > 0.1). The extent of the resection fell by 0.011 cm(3) per 1 mg/cm(3) areal bone mineral density, i.e., a slightly lower degree of bone resection was associated with a higher bone mineral density. Assuming that the resection volume was adequate to treat the patient's complaints a smaller resection volume seen in our study using an endoscopic technique might lead to fewer postoperative complaints and faster recovery.


Deactivation Kinetics of Solid Acid Catalyst with Laterally Interacting Protons.
Differences in catalyst deactivation kinetics in solid acid catalysis are studied with catalyst models that allow for lateral interaction between protons. Deactivation of a solid acid catalyst with laterally interacting protons induces inhomogeneity of proton reactivity that develops with time. As a consequence, product selectivity changes and deactivation will accelerate. This is demonstrated by simulations of the deactivation kinetics of the alkylation reaction of propylene with isobutane. The effect of lateral interactions between protons arises because initial catalyst deactivation is not caused by pore blocking or coke deposition but by a molecular mechanism where protons are consumed due to the formation of stable nonreactive carbenium ions. High selectivity to alkylate requires a catalyst with protons of high reactivity. When protons become consumed by formation of stable deactivating carbenium ions, initially reactive protons are converted into protons of lower reactivity. The latter only catalyze deactivating oligomerization reactions. Simulations that compare the deactivation kinetics of a catalyst model with laterally interacting protons and a catalyst model that contains protons of similar but different reactivity, but that do not laterally interact, are compared. These simulations demonstrate that the lateral interaction catalyst model is initially more selective but also has a lower stability. Catalyst deactivation of the alkylation reaction occurs through two reaction channels. One reaction channel is due to oligomerization of reactant propylene. The other deactivation reaction channel is initiated by deprotonation of intermediate carbenium ions that increase alkene concentration. By consecutive reactions, this also leads to deactivation. The hydride transfer reaction competes with oligomerization reactions. It is favored by strongly acid sites that also suppress the deprotonation reaction. Within the laterally interacting proton catalyst model, when reactive protons become deactivated, weakly reactive protons are generated that only catalyze the deactivating alkene oligomerization and consecutive reactions. This rapid formation of the weakly reactive protons is the cause of decreasing selectivity with reaction time and increased rate of deactivation. Solutions of the mean field kinetic equations as well as stochastic simulations are presented. Comparative simulations with a reduced number of neighbors of the protons illustrate decreased deactivation rates when the proton density decreases. Island formation of adsorbed reaction intermediates on the catalyst surface is observed in stochastic kinetics simulations. When alkylation selectivity is high, this island formation increases the rate of catalyst deactivation in comparison to the rate of deactivation according to the mean field studies. A nonlinear dynamics model of proton dynamics is provided, which shows that the differences between stochastic and mean field simulations are due to frustrated proton state percolation.


Risk Factors of Pulmonary Hypertension in Patients on Hemodialysis: A Single Center Study.
Pulmonary hypertension (PH) is an underestimated cardiovascular consequence and a mortality predictor in patients on hemodialysis (HD). Thus, we studied its prevalence, risk factors, association with inflammation/oxidative stress, and cardiac changes in HD patients. This was a single-center cross-sectional observational study conducted at a tertiary care hospital. Patients aged >18 years on hemodialysis for at least three months were included and divided into those with and without PH; patients with secondary causes for PH were excluded. Clinical characteristics, HD-related factors, lab parameters (C-reactive protein and malondialdehyde with thiol assay were used as markers of inflammation and oxidative stress, respectively), and echocardiography details were compared. PH was defined as a mean pulmonary artery pressure of >25 mmHg at rest, and it was further divided as mild (25-40 mmHg), moderate (40-60 mmHg), and severe (>60 mmHg). Of 52 patients, 28 patients had PH (mild 24, moderate 4, and none had severe PH) with prevalence of 54%. No difference was found in clinical characteristics, dialysis-related factors, biochemical parameters including inflammation (C-reactive protein; p=0.76), or oxidative stress (thiol; p=0.36 and MDA; p=0.46) between the groups. When compared to individuals without PH, HD patients with PH exhibited significantly more mitral regurgitation (p=0.002). Hemodialysis patients have a high prevalence of PH. PH was significantly associated with the presence of mitral regurgitation on echocardiography. Our study did not find differences in traditional risk factors, HD-related factors, and inflammation/oxidative markers between the groups with and without PH.


Overview of occupational epidemiology: issues/dilemmas.
Occupational epidemiology is an essential discipline in the practice of occupational medicine. Many issues/dilemmas that plague the success of the practice in this field such as absence of complete and integrated company records, loss-to-follow-up, use of records for a purpose other than that for which they were intended, variations in exposure with calendar time and arising from within plant movement, the broad ranges of response with their resultant advantages and disadvantages, the magnitude of the probability of response and its relationship to the size of the study population, have been discussed. The current approach to solving these many issues/dilemmas is by the development of health information systems that prospectively gather employee medical surveillance data, tracking of employee work history and exposure monitoring data and have the built-in ability to relate these data for health studies. Care needs to be taken to assure that comparable data are gathered at different geographic sites within a company and possibly across an industry so that small but real probabilities of response can be identified.


Microarray expression analysis of genes and pathways involved in growth plate cartilage injury responses and bony repair.
The injured growth plate cartilage is often repaired by a bone bridge which causes bone growth deformities. Whilst previous studies have identified sequential inflammatory, fibrogenic, osteogenic and bone remodelling responses involved in the repair process, the molecular pathways which regulated these cellular events remain unknown. In a rat growth plate injury model, tissue from the injury site was collected across the time-course of bone bridge formation using laser capture microdissection and was subjected to Affymetrix microarray gene expression analysis. Real Time PCR and immunohistochemical analyses were used to confirm changes in levels of expression of some genes identified in microarray. Four major functional groupings of differentially expressed genes with known roles in skeletal development were identified across the time-course of bone bridge formation, including Wnt signalling (SFRP1, SFRP4, β-catenin, Csnk2a1, Tcf7, Lef1, Fzd1, Fzd2, Wisp1 and Cpz), BMP signalling (BMP-2, BMP-6, BMP-7, Chrd, Chrdl2 and Id1), osteoblast differentiation (BMP-2, BMP-6, Chrd, Hgn, Spp1, Axin2, β-catenin, Bglap2) and skeletal development (Chrd, Mmp9, BMP-1, BMP-6, Spp1, Fgfr1 and Traf6). These studies provide insight into the molecular pathways which act cooperatively to regulate bone formation following growth plate cartilage injury and highlight potential therapeutic targets to limit bone bridge formation.


Amphidinol 3 preferentially binds to cholesterol in disordered domains and disrupts membrane phase separation.
Amphidinol 3 (AM3), a polyhydroxy-polyene metabolite from the dinoflagellate Amphidinium klebsii, possesses potent antifungal activity. AM3 is known to interact directly with membrane sterols and permeabilize membranes by forming pores. Because AM3 binds to sterols such as cholesterol and ergosterol, it can be assumed that AM3 has some impact on lipid rafts, which are membrane domains rich in sphingolipids and cholesterol. Hence, we first examined the effect of AM3 on phase-separated liposomes, in which raft-like ordered and non-raft-like disordered domains are segregated. Consequently, AM3 disrupted the phase separation at 22 μM, as in the case of methyl-β-cyclodextrin, a well-known raft-disrupter that extracts sterol from membranes. The surface plasmon resonance measurements and dye leakage assays show that AM3 preferentially recognizes cholesterol in the disordered membrane, which may reflect a weaker lipid-cholesterol interaction in disordered membrane than in ordered membrane. Finally, to gain insight into the AM3-induced coalescence of membrane phases, we measured membrane fluidity using fluorescence correlation spectroscopy, demonstrating that AM3 significantly increases the order of disordered phase. Together, AM3 preferentially binds to the disordered phase rather than the ordered phase, and enhances the order of the disordered phase, consequently blending the separated phases.


Allogeneic peripheral blood stem cell transplantation: clinical overview and nursing implications.
To provide a comprehensive understanding of allogeneic peripheral blood stem cell transplantation (PBSCT), the new modality for treating advanced hematologic malignancies. Journal articles/abstracts, published literature, clinical experience. Growth factor mobilization and apheresis is well tolerated by donors and supplies adequate numbers of stem cells for engraftment. Patients engraft sooner using PBSCT compared to bone marrow transplant (BMT). Allogeneic PBSCT is a safe alternative to BMT and has distinct advantages for donors and recipients. Faster engraftment results in fewer transfusions, shorter hospitalization, and decreased cost. Future research to determine if long-term side effects from growth factors will negatively affect donors is essential. Data regarding durability of hematopoiesis and incidence of graft versus host disease warrant further analysis. Care for patients is similar to that of standard BMT; however, care for donors is unique. Shorter hospitalization requires nurses to alter content and timing of patient education and preparation for discharge.


Effects of dietary phytate, calcium and magnesium levels on zinc bioavailability to rats.
Young rats were fed diets containing 12 mg Zn/kg and varied levels of sodium phytate for 21-day ad libitum feeding periods. In experiment 1, Ca levels were 0.3, 0.5, 0.8 and 1.0%, and phytate:Zn molar ratio varied between 0 and 50. In experiment 2, Ca was maintained at 0.3%, Mg levels were 0.07, 0.22 and 0.37%, and phytate:Zn molar ratios were 0, 10, 20 and 30 at each Mg level. Major response criteria were body weight gain and tibia Zn accumulation. Weight gain was not influenced by Ca level in the absence of phytate or by phytate at 0.3% Ca; it was increasingly depressed as phytate was increased and by each increase in Ca in the presence of phytate. Total tibia Zn content was decreased at the highest Ca level in the absence of phytate; increasing the phytate progressively depressed tibia Zn at all Ca levels. Mg and phytate additions did not affect weight gain. Tibia Zn tended to be depressed by Mg and by phytate but these effects were significant only at the highest levels of the combined additions. These data corroborate and extend previously published findings on Ca and phytate effects on Zn utilization and show bone Zn accumulation to be a more sensitive criterion than weight gain in this connection. They also indicate that Mg exerts a less pronounced effect on Zn utilization in phytate-containing diets than does Ca.


Do enzymatic analyses of serum triglycerides really need blanking for free glycerol?
Distributions of concentrations of free glycerol in clinical plasma obtained for triglyceride assay were compiled to determine the frequency with which increased concentrations of free glycerol posed a potential problem for clinical interpretation of triglyceride results. Clinical histories were studied in patients with increased concentrations of free glycerol to ascertain possible reasons for the increase and to assess the relative clinical importance of glycerol-blank-corrected triglyceride results. Significant increases in free glycerol were very uncommon, usually occurring in patients receiving glycerol-containing hyperalimentation fluids, those receiving heparin (which causes both in vivo and in vitro increases in free glycerol), or those critically ill. Free glycerol was never increased significantly in a large outpatient population. Monitoring lipid metabolism in critically ill patients, or measuring true triglyceride concentrations in patients receiving glycerol-containing fluids, may represent rare exceptions for which glycerol-blank correction is necessary for accurate clinical diagnosis and management. We conclude that there is insufficient justification for the routine expenditure of extra time and reagents to correct most analytical enzymatic triglyceride methods for free glycerol.


Interaction of VP8 with mRNAs of bovine herpesvirus-1.
The UL47 gene product, VP8, is one of the most abundant tegument proteins of bovine herpesvirus-1 (BoHV-1). Deletion of VP8 leads to impaired growth in tissue culture, and VP8 is indispensable for BHV-1 replication in cattle. To elucidate the biological functions of VP8, we explored its interaction with mRNAs of immediate early (bICP0), early (gB, gD) and late (gC) genes of BoHV-1. FLAG-tagged VP8 was pulled down from COS-7 cells co-transfected with plasmids encoding VP8 and either gB, gC, gD or bICP0. This was followed by RNA extraction, cDNA synthesis and qPCR, which demonstrated binding of VP8 to bICP0, gB, gC and gD mRNAs in the cytoplasm and nucleus. These results were supported by co-localization of VP8 with bICP0, gB, gC and gD mRNAs in the nucleus as determined by confocal microscopy. Amino acids 259-342, located in the conserved portion of UL47 homologues, were found to contain the RNA binding region on VP8. To further characterize these interactions, Northwestern blotting was performed by immobilizing VP8 on a nitrocellulose membrane followed by hybridization with in vitro transcribed bICP0 mRNA. The results demonstrated binding of VP8 to intron-less mRNA but not intron-containing mRNA of bICP0. In addition, the interaction of VP8 with bICP0 mRNA was confirmed in vitro by RNA electrophoretic mobility shift assay, which also showed that the zinc finger and acidic domains both interact with VP8. Based on these results, we concluded that VP8 binds to intron-less mRNAs of bICP0, gB, gC and gD.


Enhancing glycan isomer separations with metal ions and positive and negative polarity ion mobility spectrometry-mass spectrometry analyses.
Glycomics has become an increasingly important field of research since glycans play critical roles in biology processes ranging from molecular recognition and signaling to cellular communication. Glycans often conjugate with other biomolecules, such as proteins and lipids, and alter their properties and functions, so glycan characterization is essential for understanding the effects they have on cellular systems. However, the analysis of glycans is extremely difficult due to their complexity and structural diversity (i.e., the number and identity of monomer units, and configuration of their glycosidic linkages and connectivities). In this work, we coupled ion mobility spectrometry with mass spectrometry (IMS-MS) to characterize glycan standards and biologically important isomers of synthetic αGal-containing O-glycans including glycotopes of the protozoan parasite Trypanosoma cruzi, which is the causative agent of Chagas disease. IMS-MS results showed significant differences for the glycan structural isomers when analyzed in positive and negative polarity and complexed with different metal cations. These results suggest that specific metal ions or ion polarities could be used to target and baseline separate glycan isomers of interest with IMS-MS. Graphical abstract Glycan isomers, such as fructose and glucose, show distinct separations in positive and negative ion mode.


A volume-independent component to postdiuretic sodium retention in humans.
Net sodium (Na) loss during diuretic administration is limited by postdiuretic renal salt retention. This could be a homeostatic response to extracellular fluid volume (ECV) depletion. However, rats infused with loop diuretics develop structural and functional adaptations in the distal nephron that enhance NaCl reabsorption. Therefore, the hypothesis that postdiuretic Na retention in humans contains a volume-independent component was tested. Normal volunteers were equilibrated to a 120 mmol/24 h Na intake. For the first protocol, subjects received, in random order, a placebo, bumetanide (B), or bumetanide accompanied by an infusion of an electrolyte solution at a rate adjusted to match urine flow and thereby to obviate Na losses (bumetanide plus volume replacement; B + VR). After the completion of B diuresis, there was a positive Na balance that restored 70% of the Na loss within 42 h. However, this positive Na balance was prevented by volume replacement (B + VR). For the second protocol, subjects received, in random order, a placebo injection and a 100-mmol NaCl load (P + NaCl) or a bumetanide injection plus volume replacement in addition to a 100-mmol NaCl load (B + VR + NaCl). Over the ensuing 42 h, 94% of the load was eliminated when it was infused alone (P + NaCl). In contrast, only 9% was eliminated when it was given with bumetanide and volume replacement (B + VR + NaCl). It was concluded that postdiuretic Na retention in normal human subjects is due both to ECV depletion and to volume-independent Na retention manifest as an inability to excrete a modest NaCl load.(ABSTRACT TRUNCATED AT 250 WORDS)


[Telemedicine: expected challenges in Chile based on the experience in the United States during the pandemic].
Telemedicine has had a significant role during the outbreak of COVID-19. The experience in the United States has shown advantages and some challenges that need to be addressed to include telemedicine as an established part of the health system. This article aims to determine the United States' main challenges, associating them with the Chilean reality. In this study, we classify the barriers and difficulties for telemedicine into three areas: accessibility, cyber security, and medical liability.We argue that Chile will have to deal with similar obstacles to include telemedicine as a regular health service for the entire population.


Diurnal Variation of Optical Coherence Tomography Measurements of Static and Dynamic Anterior Segment Parameters.
To examine the diurnal variation of static and dynamic anterior segment parameters in young, healthy eyes by comparing anterior segment optical coherence tomography (AS-OCT) measurements obtained in the morning and evening and also in the light and dark. Twenty-two subjects ranging from 19 to 47 years of age with no past ocular history were selected. Imaging was performed with the Tomey CASIA2 AS-OCT device in 2 fixed lighting environments, light and dark, between the hours of 08:30 to 10:00 and 17:30 to 19:00. Four AS-OCT images were analyzed per eye. Pupil diameter (PD), iris area (IA), iris curvature (IC), anterior chamber depth (ACD), lens vault (LV), anterior chamber width (ACW), anterior chamber area (ACA), angle opening distance (AOD), angle recess area (ARA), trabecular iris space area (TISA), and trabecular iris angle (TIA) were measured. Pupil diameter was similar between the AM and PM groups in the light (P=0.89) and dark (P=0.51). There was no significant difference between AM and PM measurement values for any of the static or dynamic parameters in the light (P>0.39) and dark (P>0.31). Intraclass correlation coefficients (ICC) demonstrated excellent agreement between AM and PM measurement values in the light (ICC>0.81) and dark (ICC>0.93). In addition, there was no significant difference between AM and PM angle opening distance at 500 µm measurement values in the light (P>0.34) and dark (P>0.40) when each of 8 angle sectors was analyzed individually. No significant diurnal variation of static or dynamic anterior segment parameter measurements was detected in the light and dark. Diurnal variation of these parameters does not regularly occur in young, healthy eyes.


An evaluation of midwives' counseling of pregnant women in fear of childbirth.
The aim of this clinical evaluation was to study birth experience, post-traumatic stress symptoms, and satisfaction with care in new mothers who had consulted specially trained midwives because of a fear of childbirth during pregnancy. Sixty-two women were eligible for the study, of whom 53 (85%) participated at 1-14 months postpartum. For comparison, a group of 53 women were matched for parity and mode of delivery. All 106 women completed two self-rating scales, the Wijma Delivery Experience Questionnaire (W-DEQ) and the Impact of Event Scale (IES), and answered several open questions about their opinion of the antenatal preparation given. Those women who had been treated for fear of childbirth reported a rather more frightening experience of delivery, and more frequent symptoms of post-traumatic stress related to delivery than did the women in the comparison group. Nevertheless, satisfaction with care was manifest in the study group. Women who seek help for fear of childbirth are a vulnerable group. Because the counseling received by the women in this study did not accord them the same positive experience of childbirth as the average parturient at the unit, more effective forms of treatment may be necessary. However, as most of the women were very satisfied with their care and with the outcome, one may assume that the care given had improved their situation to some degree.


Stimulation of acid secretion increases the gastric gland luminal pressure in the rat.
The gastric mucosal gland luminal pressure was measured in vivo with a pressure-sensitive microelectrode technique (servo-null) in anesthetized rats. A microelectrode was inserted into a gland lumen by means of a micromanipulator at an angle of 30 degrees to the mucosal surface. Acid secretion was estimated by measuring the pH in the solution covering the mucosa. During control conditions, when the mucosa was secreting acid spontaneously, gland luminal pressure was 12.3 +/- 1.2 mm Hg. At about 9 minutes after starting pentagastrin administration, the luminal pressure stabilized at 17.2 +/- 1.7 mm Hg. In the rats given impromidine (500 micrograms.kg-1.h-1) luminal pressure gradually increased (during 9-10 minutes) from a control level of 9.0 +/- 1.9 to 17.3 +/- 2.6 mm Hg. During the majority of experiments, the luminal pressure oscillated at 3-7 cycles per minute. The results show that intraluminal pressure increases during stimulated acid secretion, indicating that a resistance to the volume secretion exists in the upper part of the gastric crypts. This hydrostatic pressure may well be the driving force for creating channels for acid and pepsin to cross the mucus layer covering the mucosal surface.


Algorithms for rapid outbreak detection: a research synthesis.
The threat of bioterrorism has stimulated interest in enhancing public health surveillance to detect disease outbreaks more rapidly than is currently possible. To advance research on improving the timeliness of outbreak detection, the Defense Advanced Research Project Agency sponsored the Bio-event Advanced Leading Indicator Recognition Technology (BioALIRT) project beginning in 2001. The purpose of this paper is to provide a synthesis of research on outbreak detection algorithms conducted by academic and industrial partners in the BioALIRT project. We first suggest a practical classification for outbreak detection algorithms that considers the types of information encountered in surveillance analysis. We then present a synthesis of our research according to this classification. The research conducted for this project has examined how to use spatial and other covariate information from disparate sources to improve the timeliness of outbreak detection. Our results suggest that use of spatial and other covariate information can improve outbreak detection performance. We also identified, however, methodological challenges that limited our ability to determine the benefit of using outbreak detection algorithms that operate on large volumes of data. Future research must address challenges such as forecasting expected values in high-dimensional data and generating spatial and multivariate test data sets.


[From epidemiology to social epidemiology in dentistry. The example of caries].
Today, we know the caries aetiology and prevention rather well. We also know that caries-related levels of dental health are inequitably distributed among social classes: on the average, disadvantaged people experience higher DMFS then privileged people. This difference can be explained by a differential access to health care, which can be shown by the study of components of the DMFS index: proportion of filled surfaces (F/DMFS), proportion of missing surfaces (M/DMFS), and proportion of recurrent caries (secondary caries/filled surfaces). Though we are able to describe the social distribution of levels of dental care, we cannot "explain" the differences we observe. The explanation of those differences defines the field of social epidemiology. This consists in determining the social aetiology of diseases, starting from social determinants of attitudes and behaviors influencing health.


Loop-closure kinetics reveal a stable, right-handed DNA intermediate in Cre recombination.
In Cre site-specific recombination, the synaptic intermediate is a recombinase homotetramer containing a pair of loxP DNA target sites. The enzyme system's strand-exchange mechanism proceeds via a Holliday-junction (HJ) intermediate; however, the geometry of DNA segments in the synapse has remained highly controversial. In particular, all crystallographic structures are consistent with an achiral, planar Holliday-junction (HJ) structure, whereas topological assays based on Cre-mediated knotting of plasmid DNAs are consistent with a right-handed chiral junction. We use the kinetics of loop closure involving closely spaced (131-151 bp) loxP sites to investigate the in-aqueo ensemble of conformations for the longest-lived looped DNA intermediate. Fitting the experimental site-spacing dependence of the loop-closure probability, J, to a statistical-mechanical theory of DNA looping provides evidence for substantial out-of-plane HJ distortion, which unequivocally stands in contrast to the square-planar intermediate geometry from Cre-loxP crystal structures and those of other int-superfamily recombinases. J measurements for an HJ-isomerization-deficient Cre mutant suggest that the apparent geometry of the wild-type complex is consistent with temporal averaging of right-handed and achiral structures. Our approach connects the static pictures provided by crystal structures and the natural dynamics of macromolecules in solution, thus advancing a more comprehensive dynamic analysis of large nucleoprotein structures and their mechanisms.


Switching of pyruvate kinase isoform L to M2 promotes metabolic reprogramming in hepatocarcinogenesis.
Hepatocellular carcinoma (HCC) is an aggressive tumor, with a high mortality rate due to late symptom presentation and frequent tumor recurrences and metastasis. It is also a rapidly growing tumor supported by different metabolic mechanisms; nevertheless, the biological and molecular mechanisms involved in the metabolic reprogramming in HCC are unclear. In this study, we found that pyruvate kinase M2 (PKM2) was frequently over-expressed in human HCCs and its over-expression was associated with aggressive clinicopathological features and poor prognosis of HCC patients. Furthermore, knockdown of PKM2 suppressed aerobic glycolysis and cell proliferation in HCC cell lines in vitro. Importantly, knockdown of PKM2 hampered HCC growth in both subcutaneous injection and orthotopic liver implantation models, and reduced lung metastasis in vivo. Of significance, PKM2 over-expression in human HCCs was associated with a down-regulation of a liver-specific microRNA, miR-122. We further showed that miR-122 interacted with the 3UTR of the PKM2 gene. Re-expression of miR-122 in HCC cell lines reduced PKM2 expression, decreased glucose uptake in vitro, and suppressed HCC tumor growth in vivo. Our clinical data and functional studies have revealed a novel biological mechanism involved in HCC metabolic reprogramming.


Long-term immunotherapy for perennial allergic rhinitis: relationship of specific IgG levels to skin-prick test results and clinical symptoms and signs.
We studied 63 adults with perennial allergic rhinitis and positive skin-prick tests for Dermatophagoides pteronyssinus (Dp) and Dermatophagoides farinae (Df) who had not previously received any type of immunotherapy or pharmacotherapy for their condition. We injected these patients with a 50% Dp/50% Df modified allergen intradermally in accordance with the classic immunotherapy scheme once a week for 30 months. Specific immunotherapy (SIT) was administered until the specific IgG (sIgG) concentration reached level IV (>or=81% absorbance). We measured specific IgE and sIgG levels, obtained skin-prick test results, and evaluated clinical symptoms and signs before immunotherapy, at 6 months into therapy, at the completion of therapy (30 mo), and 1 year after the completion of therapy. In the group as a whole, differences between mean sIgG values and mean skin-prick test results before and after SIT were statistically significant (p < 0.05). At the 1-year follow-up, there was a negative correlation between sIgG levels and clinical symptom and sign scores, a positive correlation between skin-prick test results and clinical improvement, and a negative correlation between sIgG levels and skin-prick test results. Individually, at the 1-year follow-up, 50 of the 63 patients (79%) had experienced a resolution of clinical symptoms and/or signs and were thus deemed to have achieved clinical recovery.


Long-term management of gastroesophageal reflux disease and its complications.
Although gastroesophageal reflux disease (GERD) is believed to be primarily a motor disorder, current medical therapy is based on the inhibition of acid secretion, since it is the deleterious effects of the acidic refluxate that lead to the symptoms and complications of GERD. Goals of long-term management include the relief of symptoms, healing of esophagitis and prevention of its relapse, and prevention of complications with safe, cost-effective therapy. Maintenance therapy depends on disease severity. Prokinetic drugs have a limited role except in symptomatic nonerosive GERD. Likewise, H2-receptor antagonists are useful in relapsing, nonerosive GERD or in cases of mild initial esophagitis. For severe reflux esophagitis, even high doses of H2-receptor antagonists do not appear to be as effective as proton pump inhibitors. GERD patients with severe reflux esophagitis or complications such as peptic stricture or Barrett's esophagus should be maintained on proton pump inhibitors such as lansoprazole or omeprazole. For young and otherwise healthy patients, antireflux surgery is a viable option.


Time trends and gender differences in incidence and prevalence of type 1 diabetes in Sweden.
There are different opinions on a possible sex bias in diabetes. In Sweden we have access to data since the 1930s, making it an ideal model. We aimed to study gender differences and time trends in the incidence and prevalence of type 1 diabetes in Sweden. We found 31 articles on incidence and 8 on prevalence (6 overlapping). Times series on incidence were found regarding children 0-15 years of age (with the Swedish Childhood Diabetes Registry, SCDR, since 1977), with up to 14,721 children with diabetes and with a high degree of ascertainment. Incidence time series were also found for subjects aged 15-34 (Diabetes Incidence Study in Sweden, DISS, since 1983), with up to 7,369 subjects and with a lower degree of ascertainment compared to SCDR. Regarding age from 40 years and above fewer studies were found, and with a much lower number of subjects with type 1 diabetes. Diabetes incidence in children has had a relative increase of approximately 2% per year since 1938. Incidence rates in children 0-14 years of age show no gender differences, but in subjects aged 15-39 years a male preponderance up to twofold is found. Figures for subjects 40 years or older are more uncertain, but show a fairly equal incidence among men and women. The male preponderance in type 1 diabetes from age 15 up to 40-50 could be due to hormonal influence, with higher peripheral insulin resistance among men in young adults and younger middle age.


Transient Expression of Human Cytochrome P450s 2D6 and 3A4 in Nicotiana benthamiana Provides a Possibility for Rapid Substrate Testing and Production of Novel Compounds.
Employment of transient expression of foreign genes for bioconversion of pharmaceutically valuable low-molecular-weight compounds, including plant secondary metabolites, is an enticing trend still scantily explored in plant biotechnology. In the present work, an efficient protocol for rapid assessment of synthetic and plant-derived metabolites as potential substrates for human P450s (CYP2D6 and CYP3A4) via Agrobacterium-mediated transient expression in Nicotiana benthamiana is put forth. Animal P450s with broad substrate specificity are promising candidates for transformation of diverse metabolites. The efficiency of P450s in heterologous surroundings is not always satisfactory and depends on the availability of an associated electron-transfer enzyme. Plants represent an attractive assortment of prospective hosts for foreign P450s expression. The optimal composition of genetic blocks providing the highest transient expression efficiency is designed, an effective substrate administration scheme is validated, and biological activity of the investigated P450s against loratadine and several indole alkaloids with different molecular scaffold structures is tested. A novel indole alkaloid, 11-hydroxycorynanthine, is isolated from N. benthamiana plants transiently expressing CYP2D6 and supplemented with corynanthine, and its structure was elucidated. The proposed technique might be of value in realization of combinatorial biosynthesis concept comprising the junction of heterologous enzymes and substrates in different metabolic surroundings.


Reversal of the behavioural effects of monocular deprivation in the kitten.
1. Eighteen kittens were monocularly deprived of vision until the age of 5, 6 or 7 weeks. Their eyes were then reverse-sutured, and they were allowed to survive for 3-63 days, before physiological recording from area 17. 2. At the time the reverse-suture was performed, and immediately before the recording session, each kitten was tested separately in the two eyes to elicit five simple behavioural responses: optokinetic nystagmus, visual startle reaction, visually-guided paw placing, visual following and negotiation of a "visual cliff". 3. Following the opening of their initially deprived eye, all kittens appeared behaviourally blind when forced to use that eye; their performance through the initially open eye was then perfect on all tests. After the period of reversed lid-suture, however, their performance when using the initially deprived eye had improved, while that through the initially open eye deteriorated. This complementary improvement and deterioration was most rapid in kittens reverse-sutured at the age of 5 weeks, and less rapid when reverse-suturing was delayed until the age of 6 or 7 weeks. 4. Most of the kittens showed gross abnormalities of interocular alignment, and exhibited marked exotropia or esotropia. 5. The results of these tests were well correlated with the changes seen in cortical ocular dominance in the same animals.


Gene network analyses support subfunctionalization hypothesis for duplicated hsp70 genes in the Antarctic clam.
A computationally predicted gene regulatory network (GRN), generated from mantle-specific gene expression profiles in the Antarctic clam Laternula elliptica, was interrogated to test the regulation and interaction of duplicated inducible hsp70 paralogues. hsp70A and hsp70B were identified in the GRN with each paralogue falling into unique submodules that were linked together by a single shared second neighbour. Annotations associated with the clusters in each submodule suggested that hsp70A primarily shares regulatory relationships with genes encoding ribosomal proteins, where it may have a role in protecting the ribosome under stress. hsp70B, on the other hand, interacted with a suite of genes involved in signalling pathways, including four transcription factors, cellular response to stress and the cytoskeleton. Given the contrasting submodules and associated annotations of the two hsp70 paralogues, the GRN analysis suggests that each gene is carrying out additional separate functions, as well as being involved in the traditional chaperone heat stress response, and therefore supports the hypothesis that subfunctionalization has occurred after gene duplication. The GRN was specifically produced from experiments investigating biomineralization; however, this study shows the utility of such data for investigating multiple questions concerning gene duplications, interactions and putative functions in a non-model species.


[Steroids' action on dependence of outset, maintenance of gravidity and onset of labor].
Dependence of outset, maintenance of gravidity and onset of labor on different kinds of steroid hormones has long been the focus of debate with the underlying control mechanism of these hormones in question remaining left unestablished still in these days. Of author's particular concern in this paper was the two-sided aspect of steroid functions. In fact, the steroid may stimulate mother's organ, fetus and placenta to produce much amount of steroid hormone throughout gravidic interval, while at the same time it may control the two contradictory processes, namely the maintenance of gravity and onset of labor through one and the same route. With a view to solve the complexity referred to above an effort has been made through a series of experiments particularity in the term of: 1. Dependent of a system capable to simultaneously determine concentrations of a variety of steroid hormones using the high performance liquid chromatography (HPLC); 2. Profile-analysis to be made on behavior of different kinds of steroids observable over gravidic interval under a support of HPLC system; 3. Behavior of different kinds of hemogenic hormones subsequent to an active DHAS loading in both group with- and without labor pains; 4. Influence of DHAS and gestation on activity of 11 beta hydroxy steroid dehydrogenase in placenta; Some of the results brought by these experiments, which are of some value to speculate on the mechanism of labor-onset and growth of fetus, may be summarized below in line with the preceding items: 1. The measuring system incorporating HPLC made it possible to determine 8 kinds of steroid hormones under systems of C19 and C21 readily in approx. 45 min., even when pretreatment is made involved in the procedure. It was also demonstrably verified that the system shows an acceptable sensitivity combined with a higher correlation with the RIA-method. 2. Profile was successfully delineated of those steroids which would participate in controlling the labor pains inducing mechanism through a series of analytical works made on the behavior of steroid associated with the process of gravidism. With the approach of expected labor, it was seen that 5 kinds of steroid hormones comprising DHA, testosterone, pregnenolone, 20 alpha OH-progesterone, and cortisol showed a marked increase. In contrast, androstenedione and progesterone begin to fall abruptly ever since several days preceding the date of labor. 3. Gain of DHA, 17 alpha OH-progesterone, due to DHAS-loading was clearly identified in the group with onset of labor pains.(ABSTRACT TRUNCATED AT 400 WORDS)


[Forecasting strategies of hepatitis B vaccination based on decision tree model].
The strategies of hepatitis B vaccination was studied based on the decision tree model. The results showed that the young, especially newborn should be given the highest priority for vaccination and the best strategy to the neonate is vaccination without screening for HBsAg to their mothers. The results also showed that the population groups who can take positive benefits (BCR greater than 1) after being vaccinated can be enlarged if decreasing the costs of the vaccination and tests for hepatitis B virus markers, and increasing the protect rates and years of vaccination. It told us that we should not only pay attention to the study for the more effective and cheaper vaccine, but manage scientifically and use reasonably as well.