Divergent Significance of Bone Mineral Density Changes in Aging Depending on Sites and Sex Revealed through Separate Analyses of Bone Mineral Content and Area.
Bone mineral density (aBMD) is equivalent to bone mineral content (BMC) divided by area. We rechecked the significance of aBMD changes in aging by examining BMC and area separately. Subjects were 1167 community-dwelling Japanese men and women, aged 40-79 years. ABMDs of femoral neck and lumbar spine were assessed by DXA twice, at 6-year intervals. The change rates of BMC and area, as well as aBMD, were calculated and described separately by the age stratum and by sex. In the femoral neck region, aBMDs were significantly decreased in all age strata by an increase in area as well as BMC loss in the same pattern in both sexes. In the lumbar spine region, aBMDs decreased until the age of 60 in women, caused by the significant BMC decrease accompanying the small area change. Very differently in men, aBMDs increased after their 50s due to BMC increase, accompanied by an area increase. Separate analyses of BMC and area change revealed that the significance of aBMD changes in aging was very divergent among sites and between sexes. This may explain in part the dissociation of aBMD change and bone strength, suggesting that we should be more cautious when interpreting the meaning of aBMD change.


[Influence of long-term contact lens wear on the cornea according to the data confocal microscopy].
The paper presents the results of a comparative study of corneal morphological changes by lifetime confocal miscroscopy in the long-term use of various types of contact lenses (CLs). Twenty-two patients (44 eyes) were examined, of them 8 patients (16 eyes) had worn soft CLs (SCLs) for 10-25 years (mean 17 years). 8 (16 eyes) had used hard CLs (HCLs) made of polymethyl metacrylate for 17-30 years (mean 23 years), 6 (12 eyes) had worn HCLs for 15 to 30 years (mean 20 years), hard gas-permeable CLs (HGCLs) over the past 5-15 years. The findings suggest that long-term wear of corrective CLs may lead to significant structural impairments in different corneal layers. The most characteristic changes are different types of epitheliopathies and the emergence of the signs of stromal degeneration with formation of deposits. According to our evidence, the likelihood of the detected changes increases with the long use of SCLs and reduces with the wear of HCLs, HGCLs in particular, which allows the authors to recommend that SCLs be changed to HGCLs after long (more than 10-15-year) contact correction.


Oculomotor and neurocognitive assessment of youth ice hockey players: baseline associations and observations after concussion.
Baseline scores on the King-Devick (K-D) Test, Immediate Post-Concussion Assessment and Cognitive Testing (ImPACT), Sport Concussion Assessment Tool 3 (SCAT3), and convergence were evaluated in youth hockey players. Worse K-D times were associated with worse ImPACT visual motor speed and reaction time. Eleven concussed athletes were retested, and there was a trend toward improved ImPACT and K-D times compared to baseline.


Smooth quantile normalization.
Between-sample normalization is a critical step in genomic data analysis to remove systematic bias and unwanted technical variation in high-throughput data. Global normalization methods are based on the assumption that observed variability in global properties is due to technical reasons and are unrelated to the biology of interest. For example, some methods correct for differences in sequencing read counts by scaling features to have similar median values across samples, but these fail to reduce other forms of unwanted technical variation. Methods such as quantile normalization transform the statistical distributions across samples to be the same and assume global differences in the distribution are induced by only technical variation. However, it remains unclear how to proceed with normalization if these assumptions are violated, for example, if there are global differences in the statistical distributions between biological conditions or groups, and external information, such as negative or control features, is not available. Here, we introduce a generalization of quantile normalization, referred to as smooth quantile normalization (qsmooth), which is based on the assumption that the statistical distribution of each sample should be the same (or have the same distributional shape) within biological groups or conditions, but allowing that they may differ between groups. We illustrate the advantages of our method on several high-throughput datasets with global differences in distributions corresponding to different biological conditions. We also perform a Monte Carlo simulation study to illustrate the bias-variance tradeoff and root mean squared error of qsmooth compared to other global normalization methods. A software implementation is available from https://github.com/stephaniehicks/qsmooth.


Insulin receptor phosphorylation may not be a prerequisite for acute insulin action.
An antiserum to the insulin receptor mimicked insulin's acute actions on glucose transport, phosphorylation of integral membrane proteins, and internalization of the insulin receptor in isolated rat adipose cells. These insulinomimetic actions of the antiserum occurred without the equivalent increase in phosphorylation of the beta subunit of the insulin receptor observed with insulin. Thus, a role of receptor phosphorylation in acute insulin action is now questioned.


Awareness of and attitude toward COVID-19 vaccination among individuals with COPD and the strategies to overcome vaccine hesitation: A mini review.
Coronavirus Disease 2019 (COVID-19) vaccines have a protective effect on individuals with chronic obstructive pulmonary disease (COPD), preventing them from developing severe illnesses and reducing the risk of hospitalization and mortality. However, the coverage rate of COVID-19 vaccination among this population is not satisfactory, which is associated with their lack of awareness of and negative attitudes toward COVID-19 vaccination, that is, vaccine hesitancy. We reviewed recent literatures on the vaccination status of COPD patients and vaccine hesitancy, described the factors related to vaccine hesitancy among COPD patients, and proposed strategies to improve the vaccine coverage, such as providing accurate and consistent vaccine information to the public, patient health education program, improving self-management capabilities, easy access to vaccination service, etc., which can hopefully help to improve patients' ability to cope with SARS-CoV-2 infection and reduce the COVID-19 related mortality.


[Changes in the cardiac hemodynamics and coronary blood flow in acute myocardial infarction due to perftoran].
One hundred and twenty patients with acute myocardium infarction, who were receiving perftoran, were analyzed within the case study. The microcirculation condition was determined by biomicroscopy of the eye conjunctiva capillaries. Changes in capillaries were evaluated by the Ditzel classification. Hemostasis was evaluated by coalugram. The condition of the myocardium was examined by electrocardiography and Echo-cardiography. The contractive ability of the myocardium was assessed by a shortening degree of the anterior-posterior size and by the ejection fraction. According to the study results, the administration of perftoran increased the number of functioning capillaries, decreased the venous congestion and reduced the diameter of arterioles and venules. Hypercoagulation was significantly decreasing. The general evaluation of the contractive ability of myocardium was satisfactory in all patients.


Caveolins: structure and function in signal transduction.
The caveolin family proteins are typically associated with microdomains that are found in the plasma membrane of numerous cells. These microdomains are referred to as/called caveolae. Caveolins are small proteins (18-24 kDa) that have a hairpin loop conformation with both the N and C termini exposed to the cytoplasm. Apart from having a structural function within caveolae, these proteins have the capacity to bind cholesterol as well as a variety of proteins, such as receptors, Src-like kinases, G-proteins, H-Ras, MEK/ERK kinases and nitric oxide synthases, which are involved in signal transduction processes. Considerable data allow the assumption to be made that the majority of the interactions with signaling molecules hold them in an inactive or repressed state. The activity of caveolins seems to be dependent on its specific post-translation modifications. It is suggested that caveolins fulfill a role in the modulation of cellular signaling cascades.


Multispectral Optoacoustic Tomography: Intra- and Interobserver Variability Using a Clinical Hybrid Approach.
Multispectral optoacoustic tomography (MSOT) represents a new imaging approach revealing functional tissue information without extrinsic contrast agents. Using a clinical combined ultrasound (US)/MSOT device, we investigated the interindividual robustness and impact of intra- and interobserver variability of MSOT values in soft tissue (muscle and subcutaneous fat) of healthy volunteers. Semiquantitative MSOT values for deoxygenated (Hb), oxygenated (HbO₂) and total hemoglobin (HbT), as well as oxygen saturation (sO₂), were calculated for both forearms in transversal and longitudinal probe orientation (n = 3, 8 measurements per subject). For intraobserver reproducibility, the same examiner investigated three subjects twice. Mean values of left vs. right forearm and transversal vs. longitudinal probe orientation were compared using an unpaired Student's t test. Bland Altmann plots with 95% limits of agreement for absolute averages and differences were calculated. Intraclass correlation coefficients (ICC 2,k) were computed for three different examiners. We obtained reproducible and consistent MSOT values with small-to-moderate deviation for muscle and subcutaneous fat tissue. Probe orientation and body side had no impact on calculated MSOT values (p > 0.05 each). Intraobserver reproducibility revealed equable mean values with small-to-moderate deviation. For muscular tissue, good ICC was obtained for sO₂. Measurements of subcutaneous tissue revealed good-to-excellent ICCs for all calculated values. Thus, in this preliminary study on healthy individuals, clinical MSOT provided consistent and reproducible functional soft tissue characterization, independent on the investigating personnel.


Experimental lung transplantation using non-penetrating vascular clips for anastomosis of the pulmonary artery.
Reliable and rapid anastomosis is critical in lung transplantation. The purpose of this study is to show the appropriateness of sutureless anastomosis of the pulmonary artery using a nonpenetrating vascular clipping device. The left lung was excised and the cranial lobe was transplanted autologously in 9 dogs. The anastomosis of the left main pulmonary artery (PA) (1 cm) was performed using a vascular clipping system. PA anastomosis was performed in 7 dogs in 8 to 13 minutes. The endovascular surface of the anastomotic site was smooth, and passage of silicone rubber through the anastomosis was good 14 or 28 days after surgery. Microscopic examination of the anastomotic sites showed excellent healing without evidence of intimal hyperplasia. The vascular clipping system facilitates anastomosis of the PA in lung transplant surgery.


Competence to Consent and Its Relationship With Cognitive Function in Patients With Schizophrenia.
Decisional capacity to consent is an emerging ethical and legal concept, and is closely related to self-determination of patients facing important medical decisions or research participations. Recently, the MacArthur Competence Assessment Tool (MacCAT), a semi-structured interview consisting of four dimensions (Understanding, Appreciation, Reasoning, and Expression of a Choice), was developed to assess the decisional capacity. Decision-making capacity in a group of patients with schizophrenia, as measured by the MacCAT, has been shown to be impaired in comparison with healthy control people. However, this does not necessarily mean the presence of impaired decisional capacity in all cases. Considering the real-world practice of obtaining informed consent from patients with schizophrenia, it is important to evaluate the relationship between psychopathological features and decisional capacity of the illness. Negative symptoms of schizophrenia have been demonstrated to be related to the ability to understand information relevant to the decision, reason rationally, and appreciate a situation and its consequences. On the other hand, positive symptoms, such as delusions and hallucinations have been an inconsistent correlate of poor capacity. Furthermore, some studies indicate that impairment of cognitive function, a core symptom of schizophrenia, could be more largely associated with decisional capacity than positive and negative symptoms. Therefore, it is reasonable to assume cognitive enhancement would enlarge the capacity to consent and promote autonomy in medical treatment and research participation in patients with schizophrenia. Further studies are warranted to elucidate this and related issues.


Acceleration of viral replication and up-regulation of cytokine levels by antimalarials: implications in malaria-endemic areas.
Antimalarial drugs are widely used in malaria endemic areas, both for chemoprophylaxis and also empirically to treat patients presenting with fever. Previously, we have reported that chloroquine enhances the severity of Semliki forest virus (SFV) and encephalomyocarditis virus infection. The studies presented herein show that a broad spectrum of antimalarial drugs augmented the replication of SFV in mice, concomitant with greater tissue damage and up-regulation of mRNA levels of various inflammatory cytokine genes, including interleukin-1 receptor antagonist (IL-1Ra), II-1alpha, IL-1beta, IL-6, IL-12p40, and interferon-gamma inducing factor. Furthermore, chloroquine enhances IL-1Ra production in RAW cells in vitro. Since IL-1Ra is known to be up-regulated in a number of viral infections, we propose that a further enhancement of its expression by antimalarials may be responsible for the increased severity of viral infection in our studies. Thus, the widespread use of antimalarials in malaria-endemic areas may predispose the population to viral infections. Further studies are in progress to delineate mechanism(s) involved in cytokine up-regulation and acceleration of viral replication.


Lung Ultrasound May Support Diagnosis and Monitoring of COVID-19 Pneumonia.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) disease (COVID-19) is characterized by severe pneumonia and/or acute respiratory distress syndrome in about 20% of infected patients. Computed tomography (CT) is the routine imaging technique for diagnosis and monitoring of COVID-19 pneumonia. Chest CT has high sensitivity for diagnosis of COVID-19, but is not universally available, requires an infected or unstable patient to be moved to the radiology unit with potential exposure of several people, necessitates proper sanification of the CT room after use and is underutilized in children and pregnant women because of concerns over radiation exposure. The increasing frequency of confirmed COVID-19 cases is striking, and new sensitive diagnostic tools are needed to guide clinical practice. Lung ultrasound (LUS) is an emerging non-invasive bedside technique that is used to diagnose interstitial lung syndrome through evaluation and quantitation of the number of B-lines, pleural irregularities and nodules or consolidations. In patients with COVID-19 pneumonia, LUS reveals a typical pattern of diffuse interstitial lung syndrome, characterized by multiple or confluent bilateral B-lines with spared areas, thickening of the pleural line with pleural line irregularity and peripheral consolidations. LUS has been found to be a promising tool for the diagnosis of COVID-19 pneumonia, and LUS findings correlate fairly with those of chest CT scan. Compared with CT, LUS has several other advantages, such as lack of exposure to radiation, bedside repeatability during follow-up, low cost and easier application in low-resource settings. Consequently, LUS may decrease utilization of conventional diagnostic imaging resources (CT scan and chest X-ray). LUS may help in early diagnosis, therapeutic decisions and follow-up monitoring of COVID-19 pneumonia, particularly in the critical care setting and in pregnant women, children and patients in areas with high rates of community transmission.


Determining the number and size of background samples derived from an area adjacent to the target sample that provide the greatest support for a POI in a target sample.
When sampling an item or surface for DNA originating from an action of interest, one is likely to collect DNA unrelated to the action of interest (background DNA). While adding to the complexity of a generated DNA profile, background DNA has been shown to aid in resolving the genotypes of contributors in a targeted sample, and where references of donors to the background DNA are not available, strengthen the LR supporting a person of interest contributing to the targeted sample. This is possible thanks to advances in probabilistic genotyping, where forensic labs are able to deconvolute complex DNA profiles to obtain lists of genotypes and their associated weights. Coupled with DBLR™, one can then compare multiple evidentiary profiles to each other to determine the contribution of common, but unknown, contributors. Here, we consider factors associated with taking background samples and whether one should collect multiple background samples that all relate to a single target sample, or if one should collect larger background samples rather than smaller samples. Background samples consisted of DNA accumulated on the items primarily by one or both occupants of a single household, while targeted samples were generated from touch deposits, or saliva deposits that had been left to air dry. Samples were collected from areas of various sizes, consisting of only the background, the target and the background directly beneath it, and the target and additional surrounding background. A broad range of DNA quantities were recovered, with larger background samples (400 cm2) yielding significantly more DNA than smaller background samples (30 cm2). Significant differences in DNA quantities between target samples were not observed. Generated DNA profiles were interpreted using STRmix™ and DBLR™, and where there was support for a common donor between the background and target sample, pairwise comparisons were performed to observe the effect on the LR supporting the target DNA donor contributing to the targeted sample when conditioning on one (or two) common donor between the targeted sample and 1-8 background samples. Multiple background samples gave significantly higher LRs compared to a single background sample, the larger sampled background area resulted in larger LR gains than the smaller areas, and four or more background samples reduced LR variability considerably. Here we provide recommendations for the minimum and ideal number of additional background samples that should be collected, and that several smaller samples may be more beneficial than a single larger sample.


Reorganization energies and spectral densities for electron transfer problems in charge transport materials.
In describing the dynamics of electron transfer or charge transport, the reorganization energy and the spectral density function describe the influence of nuclei motion to the transporting electron. The spectral density can be obtained using various theoretical approaches: from a model of dielectric response, or calculated with various computational means. With the vast advancement of modern computational techniques, many details in vibronic coupling can be obtained, including those described in the early literature. In this work, we provide a comprehensive understanding of the nature of vibronic coupling in light of some early literature. The theoretical connection among different quantities for vibronic coupling is discussed, followed by a brief review of the spectral density function. Various approaches and some of the results for the spectral density function are also reviewed. The importance of low-frequency bands in nonpolar systems that can be overlooked is also discussed, for both Holstein and Peierls types of electron-phonon couplings.


Do Cardiovascular Risk Factors and Cardiovascular Disease Explain Sex Differences in Cognitive Functioning in Old Age?
Sex differences in cognitive functioning in old age are known to exist yet are still poorly understood. This study examines to what extent differences in cardiovascular risk factors and cardiovascular disease between men and women explain sex differences in cognitive functioning. Data from 2,724 older adults from the Longitudinal Aging Study Amsterdam were used. Information processing speed and episodic memory, measured three times during six years of follow-up, served as outcomes. The mediating role of cardiovascular risk factors and cardiovascular disease was examined in single and multiple mediator models. Determinant-mediator effects were estimated using linear or logistic regression, and determinant-outcome and mediator-outcome effects were estimated using linear mixed models. Indirect effects were estimated using the product-of-coefficients estimator. Women scored 1.58 points higher on information processing speed and 1.53 points higher on episodic memory. Several cardiovascular risk factors had small mediating effects. The sex difference in information processing speed was mediated by smoking, depressive symptoms, obesity, and systolic blood pressure. The sex difference in episodic memory was mediated by smoking, physical activity, and depressive symptoms. Effects of smoking, LDL cholesterol, and diabetes mellitus on information processing speed differed between men and women. Differences in cardiovascular risk factors between women and men partially explained why women had better cognitive functioning. A healthy cardiovascular lifestyle seems beneficial for cognition and sex-specific strategies may be important to preserve cognitive functioning at older age.


Inflammation in Depression and the Potential for Anti-Inflammatory Treatment.
Accumulating evidence supports an association between depression and inflammatory processes, a connection that seems to be bidirectional. Clinical trials have indicated antidepressant treatment effects for anti-inflammatory agents, both as add-on treatment and as monotherapy. In particular, nonsteroidal anti-inflammatory drugs (NSAIDs) and cytokine-inhibitors have shown antidepressant treatment effects compared to placebo, but also statins, poly-unsaturated fatty acids, pioglitazone, minocycline, modafinil, and corticosteroids may yield antidepressant treatment effects. However, the complexity of the inflammatory cascade, limited clinical evidence, and the risk for side effects stress cautiousness before clinical application. Thus, despite proof-of-concept studies of anti-inflammatory treatment effects in depression, important challenges remain to be investigated. Within this paper, we review the association between inflammation and depression together with the current evidence on use of anti-inflammatory treatment in depression. Based on this, we address the questions and challenges that seem most important and relevant to future studies, such as timing, most effective treatment lengths and identification of subgroups of patients potentially responding better to different anti-inflammatory treatment regimens.


Changes in mass and performance in rabbit muscles after muscle damage with or without transplantation of primary satellite cells.
Changes in morphology, metabolism, myosin heavy chain gene expression, and functional performances in damaged rabbit muscles with or without transplantation of primary satellite cells were investigated. For this purpose, we damaged bilaterally the fast muscle tibialis anterior (TA) with either 1.5 or 2.6 ml cardiotoxin 10(-5) M injections. Primary cultures of satellite cells were autotransplanted unilaterally 5 days after muscle degeneration. Two months postoperation, the masses of damaged TAs, with or without transplantation, were significantly larger than those of the controls. Furthermore, damaged transplanted muscles weighed significantly more than damaged muscles only. The increase in muscle mass was essentially due to increased fiber size. These results were independent of the quantity of cardiotoxin injected into the muscles. Maximal forces were similar in control and 2.6 ml damaged TAs with or without satellite cell transfer. In contrast, 1.5 ml damaged TAs showed a significant decrease in maximal forces that reached the level of controls after transplantation of satellite cells. Fatigue resistance was similar in control and 1.5 ml damaged TAs independently of satellite cell transfer. Fatigue index was significantly higher in 2.6 ml damaged muscles with or without cell transplantation. These changes could be explained in part by muscle metabolism, which shifted towards oxidative activities, and by gene expression of myosin heavy chain isoforms, which presented an increase in type IIa and a decrease in type I and IIb in all damaged muscles with or without cell transfer. Under our experimental conditions, these results show that muscle damage rather than satellite cell transplantation changes muscle metabolism, myosin heavy chain isoform gene expression, and, to a lesser extent, muscle contractile properties. In contrast, muscle weight and fiber size are increased both by muscle damage and by satellite cell transfer.


Sub-families of alpha/beta barrel enzymes: a new adenine deaminase family.
No gene coding for an adenine deaminase has been described in eukaryotes. However, physiological and genetical evidence indicates that adenine deaminases are present in the ascomycetes. We have cloned and characterised the genes coding for the adenine deaminases of Aspergillus nidulans, Saccharomyces cerevisiae and Schizosaccharomyces pombe. The A.nidulans gene was expressed in Escherichia coli and the purified enzyme shows adenine but not adenosine deaminase activity. The open reading frames coded by the three genes are very similar and obviously related to the bacterial and eukaryotic adenosine deaminases rather than to the bacterial adenine deaminases. The latter are related to allantoinases, ureases and dihydroorotases. The fungal adenine deaminases and the homologous adenosine deaminases differ in a number of residues, some of these being clearly involved in substrate specificity. Other prokaryotic enzymes in the database, while clearly related to the above, do not fit into either sub-class, and may even have a different specificity. These results imply that adenine deaminases have appeared twice in the course of evolution, from different ancestral enzymes constructed both around the alpha/beta barrel scaffold.


[Lung infarcts detected with ultrasonography].
33 patients with clinical signs of pulmonary infarction were investigated by intercostal chest sonography. 40 preponderant triangular sharply outlined echo-poor lesions were detected with an airdense reflex in the centre of the area as a sign of segmental involvement. Sonomorphology of pulmonary infarction is compared to pneumonias (n = 22) and carcinomas (n = 20). The sensitivity of chest sonography in pulmonary infarction is 96%, whereas the diagnostic accuracy is 82%.


Chaperone-Mediated Autophagy in Neurodegenerative Diseases and Acute Neurological Insults in the Central Nervous System.
Autophagy is an important function that mediates the degradation of intracellular proteins and organelles. Chaperone-mediated autophagy (CMA) degrades selected proteins and has a crucial role in cellular proteostasis under various physiological and pathological conditions. CMA dysfunction leads to the accumulation of toxic protein aggregates in the central nervous system (CNS) and is involved in the pathogenic process of neurodegenerative diseases, including Parkinson's disease and Alzheimer's disease. Previous studies have suggested that the activation of CMA to degrade aberrant proteins can provide a neuroprotective effect in the CNS. Recent studies have shown that CMA activity is upregulated in damaged neural tissue following acute neurological insults, such as cerebral infarction, traumatic brain injury, and spinal cord injury. It has been also suggested that various protein degradation mechanisms are important for removing toxic aberrant proteins associated with secondary damage after acute neurological insults in the CNS. Therefore, enhancing the CMA pathway may induce neuroprotective effects not only in neurogenerative diseases but also in acute neurological insults. We herein review current knowledge concerning the biological mechanisms involved in CMA and highlight the role of CMA in neurodegenerative diseases and acute neurological insults. We also discuss the possibility of developing CMA-targeted therapeutic strategies for effective treatments.


The Association between cutaneous squamous cell carcinoma and betapapillomavirus seropositivity: a cohort study.
It is currently unclear whether betapapillomaviruses (betaPV) play a role in the etiology of cutaneous squamous cell carcinoma (SCC). We investigated the association between betaPV antibodies and subsequent SCC in a population-based cohort study. Serum samples were collected in 1992 and/or 1996 from 1,311 participants of the community-based Nambour Skin Cancer Study. These were tested for the presence of L1 antibodies against 21 different betaPV types. Histologically diagnosed SCCs were ascertained through three full-body skin examinations and linkage with the local pathology laboratories. We used age- and sex-adjusted Cox proportional hazards models to analyze the relationship between betaPV antibodies and SCC occurrence from 1992 until 2007. SCC was newly diagnosed in 150 people. No associations were found between the presence of any betaPV L1 antibodies and the occurrence of SCC (HR = 1.0), and stratification by sex, skin color, and sunburn propensity did not affect these results. However, among people who were less than 50 years old in 1992, the presence of betaPV antibodies was associated with a two-fold increased risk of SCC. There was no significant association between antibodies to any individual betaPV type examined and the later development of SCC. Whether betaPV infection of the skin, and indirectly betaPV antibodies, are involved in the oncogenic process in the general population remains unclear, and this longitudinal study provides only limited support. This study emphasizes the need for additional longitudinal studies of HPV (human papilloma virus) and SCC, to avoid the possibility of reverse causality in case--control studies.


[Genetic map of the transposable phage D3112 of Pseudomonas aeruginosa].
Using a large group of newly isolated deletion mutants of prophage D3112 the location of all known mutations of D3112 phage was more precisely defined. The mutations affecting establishment of lysogenic state were mapped in two regions of the genome- 0-1.3 and 29-30 kb. The replicative A gene is mapped between 1.3 and 4.9 kb, the second replicative B gene being situated on the right of the A gene, between 4.9 and 9.4 kb. The C gene which is responsible for positive regulation of phage late genes' expression is mapped within the 9-12 kb region. It is suggested that promoter of the gene C is situated within the same interval. Mutations were isolated in the Lys gene which is responsible for host cell lysis. The gene is located within the interval 14-22 kb of the physical map. The order of morphogenetic genes in the late genome region was also established.


[Regulation of melanogenesis in the dyschromia of skin].
To examine the role of immune, biochemical and hormonal factors in the regulation of melanogenesis in patients with chromatopathy. We observed 226 patients with various forms dyschromia skin. Age of the patients was in the range of 16 to 55 years. The frequency of females (n = 157) prevailed over the male sex (n = 69) 2,3 times. The disease duration ranged from 3 weeks to 12 years. For the study of melanogenesis in vitiligo, nevi and melasma were studied parameters of the immune, endocrine and lipid peroxidation--antioxidant system. Melanocytes are responsible for the change in concentration a-chromatophorotropic hormone and adrenocorticotropic hormone (ACTH) decrease or increase melanogenesis. Activation process is also associated with UFOs. Comparison of the level of system components lipid peroxidation (LPO)--antioxidant system (AOS) suppressor (CD8 +) lymphocyte activity vitiligo patients showed that the most pronounced suppressive effect was observed in patients with high levels of lipid peroxidation. At the same time, patients with hyperpigmentation found a significant negative relationship with CD4+. It should be noted a negative relationship CD16(+)-lymphocytes with indicators pituitary- adrenal axis in patients Dyschromias (r = -0,318 vitiligo, r = -0,512, r = -0,4578--in nevi and melasma, respectively). The results showed that vitiliginozny process, especially actively expressed, proceeds with the intensification of lipid peroxidation processes, changes in the state of AOS and immunity. With the increased level of hyperpigmentation CD95(+)-cells led to a weakening of apoptosis and cause increase in the number of melanocytes. When there is insufficient apoptosis hyperpigmentation, hypopigmentation and when--excessive apoptosis. To find mechanisms regulating skin pigmentation necessary to determine a-chromatophorotropic hormone, adrenocorticotropic hormone and neutral endopeptidase.


Monitoring of tumor growth and post-irradiation recurrence in a diffuse intrinsic pontine glioma mouse model.
Diffuse intrinsic pontine glioma (DIPG) is a fatal malignancy because of its diffuse infiltrative growth pattern. Translational research suffers from the lack of a representative DIPG animal model. Hence, human E98 glioma cells were stereotactically injected into the pons of nude mice. The E98 DIPG tumors presented a strikingly similar histhopathology to autopsy material of a DIPG patient, including diffuse and perivascular growth, brainstem- and supratentorial invasiveness and leptomeningeal growth. Magnetic resonance imaging (MRI) was effectively employed to image the E98 DIPG tumor. [(18) F] 3'-deoxy-3'-[(18) F]fluorothymidine (FLT) positron emission tomography (PET) imaging was applied to assess the subcutaneous (s.c.) E98 tumor proliferation status but no orthotopic DIPG activity could be visualized. Next, E98 cells were cultured in vitro and engineered to express firefly luciferase and mCherry (E98-Fluc-mCherry). These cultured E98-Fluc-mCherry cells developed focal pontine glioma when injected into the pons directly. However, the diffuse E98 DIPG infiltrative phenotype was restored when cells were injected into the pons immediately after an intermediate s.c. passage. The diffuse E98-Fluc-mCherry model was subsequently used to test escalating doses of irradiation, applying the bioluminescent Fluc signal to monitor tumor recurrence over time. Altogether, we here describe an accurate DIPG mouse model that can be of clinical relevance for testing experimental therapeutics in vivo.


Reversal of long-term potentiation-like plasticity processes after motor learning disrupts skill retention.
Plasticity of synaptic connections in the primary motor cortex (M1) is thought to play an essential role in learning and memory. Human and animal studies have shown that motor learning results in long-term potentiation (LTP)-like plasticity processes, namely potentiation of M1 and a temporary occlusion of additional LTP-like plasticity. Moreover, biochemical processes essential for LTP are also crucial for certain types of motor learning and memory. Thus, it has been speculated that the occlusion of LTP-like plasticity after learning, indicative of how much LTP was used to learn, is essential for retention. Here we provide supporting evidence of it in humans. Induction of LTP-like plasticity can be abolished using a depotentiation protocol (DePo) consisting of brief continuous theta burst stimulation. We used transcranial magnetic stimulation to assess whether application of DePo over M1 after motor learning affected (1) occlusion of LTP-like plasticity and (2) retention of motor skill learning. We found that the magnitude of motor memory retention is proportional to the magnitude of occlusion of LTP-like plasticity. Moreover, DePo stimulation over M1, but not over a control site, reversed the occlusion of LTP-like plasticity induced by motor learning and disrupted skill retention relative to control subjects. Altogether, these results provide evidence of a link between occlusion of LTP-like plasticity and retention and that this measure could be used as a biomarker to predict retention. Importantly, attempts to reverse the occlusion of LTP-like plasticity after motor learning comes with the cost of reducing retention of motor learning.


Beta 1- and beta 2-receptors are differentially desensitized in an experimental model of heart failure.
Recent reports suggested that a complex alteration in beta-receptor function occurs in failing human myocardium. We evaluated beta-receptor-subtype activity in an experimental model of monocrotaline (MCT)-induced cardiomyopathy in the rat. Through pulmonary hypertension, MCT causes right ventricular hypertrophy (RVH), either associated with heart failure or not, beta-Receptor function was evaluated in both failing-hypertrophic and hypertrophic hearts in binding studies with [125I]iodocyanopindolol (ICYP) and by measuring adenylate cyclase (AC) activity. In the right failing ventricle, beta 1- but not beta 2-receptor density was decreased. Lesion-associated modifications in the adenylate cyclase system were also observed: isoproterenol- and guanosine 5' [beta, gamma-imido]triphosphate [Gpp(NH)p]-stimulated cyclic AMP formation was reduced in the right failing ventricle, while the cyclic AMP responses to NaF and forskolin were unchanged. On the other hand, no changes in either beta-receptor density or function were found in hypertrophic ventricles. MCT-induced heart failure in the rat is thus associated with a selective decrease of beta 1-receptor density and function. These results suggest that MCT-induced cardiac failure may be an appropriate model in which to investigate heart insufficiency further.


Effects of colchicine and vinblastine on output of proteins into bile.
The microtubule poisons colchicine and vinblastine caused a reduction in the biliary output of total protein, compared with controls, in bile-fistula rats. The various protein components of bile showed patterns of change in output differing from one another. Alkaline phosphodiesterase I, a 'plasma-membrane' enzyme, showed a decline in output during the first hour after drug administration. Immunoglobulin A output did not decline until after the first hour. In contrast with these reductions, the biliary output of (rat) plasma albumin into bile was increased. At no time was there any evidence (as measured by lactate dehydrogenase release into bile) for any cytolytic damage. These results are discussed in relation to current theories on the output of proteins into bile and the repair of the plasma membrane in the bile canaliculus.


[A comparative electron microscopic study of the peripheral blood leukocytes of vertebrates].
The ultrastructure of leukocytes of the peripheral blood was studied in representatives of classes of fishes (Cyprinus carpio), amphibia (Rana redibunda, Bufo bufo), reptiles (Testudo horshfield), birds (Columba livia) and mammals (Rattus norvegicus). The granules are characterized by homogeneous or heterogeneous matrix. The granules of special leukocytes (turtle) and pseudoesinophils (pigeon) have larger dimensions than those in neutrophils of other animals. The matrix of the eosinophil granules of the carp and rat contain crystalloid structures while the granules of eosinophils of amphibia, turtles and pigeons are characterized by electron dense homogeneous matrix. The basophil granules are larger and in addition to homogeneous or fine granular structure (turtle, pigeon, rat) have lamellar structure (Cyprinus carpio, frog, toad). The ultrastructural organization of agranulocytes and plasma cells of the animals studied is also described.


Advancing functional connectivity research from association to causation.
Cognition and behavior emerge from brain network interactions, such that investigating causal interactions should be central to the study of brain function. Approaches that characterize statistical associations among neural time series-functional connectivity (FC) methods-are likely a good starting point for estimating brain network interactions. Yet only a subset of FC methods ('effective connectivity') is explicitly designed to infer causal interactions from statistical associations. Here we incorporate best practices from diverse areas of FC research to illustrate how FC methods can be refined to improve inferences about neural mechanisms, with properties of causal neural interactions as a common ontology to facilitate cumulative progress across FC approaches. We further demonstrate how the most common FC measures (correlation and coherence) reduce the set of likely causal models, facilitating causal inferences despite major limitations. Alternative FC measures are suggested to immediately start improving causal inferences beyond these common FC measures.


Mrad9 and atm haploinsufficiency enhance spontaneous and X-ray-induced cataractogenesis in mice.
Rad9 and Atm regulate multiple cellular responses to DNA damage, including cell cycle checkpoints, DNA repair and apoptosis. However, the impact of dual heterozygosity for Atm and Rad9 is unknown. Using 50 cGy of X rays as an environmental insult and cataractogenesis as an end point, this study examined the effect of heterozygosity for one or both genes in mice. Posterior subcapsular cataracts, characteristic of radiation exposure, developed earlier in X-irradiated double heterozygotes than in single heterozygotes, which were more prone to cataractogenesis than wild-type controls. Cataract onset time and progression in single or double heterozygotes were accelerated even in unirradiated eyes. These findings indicate that the cataractogenic effect of combined heterozygosity is greater than for each gene alone and are the first to demonstrate the impact of multiple haploinsufficiency on radiation effects in an intact mammal. These observations may help explain observed interindividual differential radiosensitivity in human populations and have important implications for those undergoing radiotherapy or exposed to elevated levels of cosmic radiation, such as the astronaut corps. These findings demonstrate that Mrad9 and Atm are important determinants of lens opacification and, given the roles of Atm and Rad9 in maintaining genomic stability, are consistent with a genotoxic basis for radiation cataractogenesis.


Changes in mood, depression and suicidal ideation after commencing pregabalin for neuropathic pain.
Pregabalin is a treatment option for patients with persistent neuropathic pain. Its use has been associated with changes in mood and the development of depression and/or suicidal ideation. Case presentations were reviewed of five patients reporting changes in mood, depression and suicidal ideation from the first 50 (approximately) patients commenced on pregabalin at the clinic. Although these patients had a history of depression, their mood had been stable before commencing pregabalin. Soon after commencement they reported changes in mood, and development of depression and/or suicidal ideation, which improved with dose reduction or cessation of pregabalin. Ultimately, all five patients ceased pregabalin treatment. Suicidal ideation is a recognised adverse effect of pregabalin. Patients should be warned of and monitored for deterioration in mood.


Worksite follow-up and engagement strategies for initiating health risk behavior changes.
Information is presented from a multiplant study of interventions to improve cardiovascular health among employees. Risk factors targeted were high blood pressure, obesity, and cigarette smoking. The study utilized on-site wellness counselors who periodically contacted all employees identified through screening as having one or more of the three risks. Use of a structured protocol for client outreach resulted in the large majority of clients being seen in follow-up during the three-year intervention period. Drawing from caseload experience and from various theoretical perspectives, seven engagement strategies were used to help guide at-risk clients toward successful behavior changes to reduce health risks. Results showed that of the three at-risk groups, clients with high blood pressure were most likely to be seen in follow-up, and most likely to begin a risk reduction program. For the overweight and smokers, clients seen three or more times were more likely to begin a weight-loss/smoking cessation program than clients seen less often. Frequency of follow-up showed a positive relationship with risk reduction for all three risks, in samples of employees rescreened at the end of the intervention period, but the relationship was not statistically significant for smoking cessation.


Family-Centered Management of Sensory Challenges of Children With Autism: Single-Case Experimental Design.
We explored the effectiveness of a sensory-based, family-centered coaching approach to changing problematic routines for young children with autism. Three mothers of young children with autism, atypical sensory processing, and global developmental delay each participated in a single-case experimental ABA design study. Mothers selected a problematic daily routine linked to sensory challenges as the focus of four intervention sessions provided in the home. Changes in mothers' perceptions of the children's behavior were the primary outcome, measured daily on a visual analog scale. Visual and descriptive analyses were undertaken. The sensory-based, family-centered coaching approach showed promise for changing sensory-related problem behaviors of young children with autism, but the degree and maintenance of the intervention effect varied among participants.


Computational alanine scanning of protein-protein interfaces.
Protein-protein interactions are key components of all signal transduction processes, so methods to alter these interactions promise to become important tools in dissecting function of connectivities in these networks. We have developed a fast computational approach for the prediction of energetically important amino acid residues in protein-protein interfaces (available at http://robetta.bakerlab.org/alaninescan), which we, following Peter Kollman, have termed "computational alanine scanning." The input consists of a three-dimensional structure of a protein-protein complex; output is a list of "hot spots," or amino acid side chains that are predicted to significantly destabilize the interface when mutated to alanine, analogous to the results of experimental alanine-scanning mutagenesis. 79% of hot spots and 68% of neutral residues were correctly predicted in a test of 233 mutations in 19 protein-protein complexes. A single interface can be analyzed in minutes. The computational methodology has been validated by the successful design of protein interfaces with new specificity and activity, and has yielded new insights into the mechanisms of receptor specificity and promiscuity in biological systems.


Immune and Genetic Correlates of Vaccine Protection Against Mucosal Infection by SIV in Monkeys.
The RV144 vaccine trial in Thailand demonstrated that an HIV vaccine could prevent infection in humans and highlights the importance of understanding protective immunity against HIV. We used a nonhuman primate model to define immune and genetic mechanisms of protection against mucosal infection by the simian immunodeficiency virus (SIV). A plasmid DNA prime/recombinant adenovirus serotype 5 (rAd5) boost vaccine regimen was evaluated for its ability to protect monkeys from infection by SIVmac251 or SIVsmE660 isolates after repeat intrarectal challenges. Although this prime-boost vaccine regimen failed to protect against SIVmac251 infection, 50% of vaccinated monkeys were protected from infection with SIVsmE660. Among SIVsmE660-infected animals, there was about a one-log reduction in peak plasma virus RNA in monkeys expressing the major histocompatibility complex class I allele Mamu-A*01, implicating cytotoxic T lymphocytes in the control of SIV replication once infection is established. Among Mamu-A*01-negative monkeys challenged with SIVsmE660, no CD8(+) T cell response or innate immune response was associated with protection against virus acquisition. However, low levels of neutralizing antibodies and an envelope-specific CD4(+) T cell response were associated with vaccine protection in these monkeys. Moreover, monkeys that expressed two TRIM5 alleles that restrict SIV replication were more likely to be protected from infection than monkeys that expressed at least one permissive TRIM5 allele. This study begins to elucidate the mechanisms of vaccine protection against immunodeficiency viruses and highlights the need to analyze these immune and genetic correlates of protection in future trials of HIV vaccine strategies.


CREB phosphorylation at Ser133 regulates transcription via distinct mechanisms downstream of cAMP and MAPK signalling.
CREB (cAMP-response-element-binding protein) is an important transcription factor for the activation of a number of immediate early genes. CREB is phosphorylated on Ser133 by PKA (protein kinase A), promoting the recruitment of the co-activator proteins CBP (CREB-binding protein) and p300; this has been proposed to increase the transcription of CREB-dependent genes. CREB is also phosphorylated on Ser133 by MSK1/2 (mitogen- and stress-activated kinase 1/2) in cells in response to the activation of MAPK (mitogen-activated protein kinase) signalling; however, the relevance of this to gene transcription has been controversial. To resolve this problem, we created a mouse with a Ser133 to alanine residue mutation in the endogenous Creb gene. Unlike the total CREB knockout, which is perinatally lethal, these mice were viable, but born at less than the expected Mendelian frequency on a C57Bl/6 background. Using embryonic fibroblasts from the S133A-knockin mice we show in the present study that Ser133 phosphorylation downstream of PKA is required for CBP/p300 recruitment. The requirement of Ser133 phosphorylation for the PKA-mediated induction of CREB-dependent genes was, however, promoter-specific. Furthermore, we show that in cells the phosphorylation of CREB on Ser133 by MSKs does not promote strong recruitment of CBP or p300. Despite this, MSK-mediated CREB phosphorylation is critical for the induction of CREB-dependent genes downstream of MAPK signalling.


Induced Heteroresistance in Carbapenem-Resistant Acinetobacter baumannii (CRAB) via Exposure to Human Pleural Fluid (HPF) and Its Impact on Cefiderocol Susceptibility.
Infections caused by Carbapenem-resistant Acinetobacter baumannii (CRAB) isolates, such as hospital-acquired pneumonia (HAP), bacteremia, and skin and soft tissue infections, among others, are particularly challenging to treat. Cefiderocol, a chlorocatechol-substituted siderophore antibiotic, was approved by the U.S. Food and Drug Administration (FDA) in 2019 and prescribed for the treatment of CRAB infections. Despite the initial positive treatment outcomes with this antimicrobial, recent studies reported a higher-than-average all-cause mortality rate in patients treated with cefiderocol compared to the best available therapy. The cause(s) behind these outcomes remains unconfirmed. A plausible hypothesis is heteroresistance, a phenotype characterized by the survival of a small proportion of cells in a population that is seemingly isogenic. Recent results have demonstrated that the addition of human fluids to CRAB cultures leads to cefiderocol heteroresistance. Here, we describe the molecular and phenotypic analyses of CRAB heteroresistant bacterial subpopulations to better understand the nature of the less-than-expected successful outcomes after cefiderocol treatment. Isolation of heteroresistant variants of the CRAB strain AMA40 was carried out in cultures supplemented with cefiderocol and human pleural fluid (HPF). Two AMA40 variants, AMA40 IHC1 and IHC2, were resistant to cefiderocol. To identify mutations and gene expression changes associated with cefiderocol heteroresistance, we subjected these variants to whole genome sequencing and global transcriptional analysis. We then assessed the impact of these mutations on the pharmacodynamic activity of cefiderocol via susceptibility testing, EDTA and boronic acid inhibition analysis, biofilm formation, and static time-kill assays. Heteroresistant variants AMA40 IHC1 and AMA40 IHC2 have 53 chromosomal mutations, of which 40 are common to both strains. None of the mutations occurred in genes associated with high affinity iron-uptake systems or β-lactam resistance. However, transcriptional analyses demonstrated significant modifications in levels of expression of genes associated with iron-uptake systems or β-lactam resistance. The blaNDM-1 and blaADC-2, as well as various iron-uptake system genes, were expressed at higher levels than the parental strain. On the other hand, the carO and ompA genes' expression was reduced. One of the mutations common to both heteroresistant strains was mapped within ppiA, a gene associated with iron homeostasis in other species. Static time-kill assays demonstrated that supplementing cation-adjusted Mueller-Hinton broth with human serum albumin (HAS), the main protein component of HPF, considerably reduced cefiderocol killing activity for all three strains tested. Notably, collateral resistance to amikacin was observed in both variants. We conclude that exposing CRAB to fluids with high HSA concentrations facilitates the rise of heteroresistance associated with point mutations and transcriptional upregulation of genes coding for β-lactamases and biofilm formation. The findings from this study hold significant implications for understanding the emergence of CRAB resistance mechanisms against cefiderocol treatment. This understanding is vital for the development of treatment guidelines that can effectively address the challenges posed by CRAB infections.


State of disparities in cardiovascular health in the United States.
Reducing health disparities remains a major public health challenge in the United States. Having timely access to current data on disparities is important for policy and program development. Accordingly, we assessed the current magnitude of disparities in cardiovascular disease (CVD) and its risk factors in the United States. Using national surveys, we determined CVD and risk factor prevalence and indexes of morbidity, mortality, and overall quality of life in adults > or =18 years of age by race/ethnicity, sex, education level, socioeconomic status, and geographic location. Disparities were common in all risk factors examined. In men, the highest prevalence of obesity (29.2%) was found in Mexican Americans who had completed a high school education. Black women with or without a high school education had a high prevalence of obesity (47.3%). Hypertension prevalence was high among blacks (39.8%) regardless of sex or educational status. Hypercholesterolemia was high among white and Mexican American men and white women in both groups of educational status. Ischemic heart disease and stroke were inversely related to education, income, and poverty status. Hospitalization was greater in men for total heart disease and acute myocardial infarction but greater in women for congestive heart failure and stroke. Among Medicare enrollees, congestive heart failure hospitalization was higher in blacks, Hispanics, and American Indians/Alaska Natives than among whites, and stroke hospitalization was highest in blacks. Hospitalizations for congestive heart failure and stroke were highest in the southeastern United States. Life expectancy remains higher in women than men and higher in whites than blacks by approximately 5 years. CVD mortality at all ages tended to be highest in blacks. Disparities in CVD and related risk factors remain pervasive. The data presented here can be invaluable for policy development and in the planning, implementation, and evaluation of interventions designed to eliminate health disparities.


From the bench to practice - Field integration of community-based services for older citizens with different levels of functional limitation across European Regions.
The meeting of the European Innovation Partnership on Active and Healthy Ageing (EIPonAHA) action group A3 together with members of the Reference site collaborative network (RSCN) in December 2019 in Rome focused on integration of evidence-based approaches on health and care delivery for older citizens at different levels of needs with expertise coming from stakeholder across Europe. It was the final aim of the group to co-create culturally sensitive pathways and facilitate co-ownership for further implementation of the pathways in different care systems across Europe. The study design is a mixed method approach. Based on data analysis from a cohort of community-dwelling over-65 citizens in the framework of a longitudinal observational study in Rome, which included health, social and functional capacity data, three personas profiles were developed: the pre-frail, the frail and the very frail personas. Based on these data, experts were asked to co-create care pathways due to evidence and eminence during a workshop and included into a final report. All working groups agreed on a common understanding that integration of care means person-centered integration of health and social care, longitudinally provided across primary and secondary health care including citizens' individual social, economic and human resources. Elements for consideration during care for pre-frail people are loneliness and social isolation, which, lead to limitation of physical autonomy in the light of reduced access to social support. Frail people need adaption of environmental structures and, again, social resource allocation to maintain at home. Very frail are generally vulnerable patients with complex needs. Most of them remain at home because of a strong individual social support and integrated health care delivery. The approach described in this publication may represent a first approach to scaling-up care delivery in a person-centered approach.


Effect of Clinical Decision Support on Diagnostic Imaging for Pediatric Appendicitis: A Cluster Randomized Trial.
Appendicitis is the most common pediatric surgical emergency. Efforts to improve efficiency and quality of care have increased reliance on computed tomography (CT) and ultrasonography (US) in children with suspected appendicitis. To evaluate the effectiveness of an electronic health record-linked clinical decision support intervention, AppyCDS, on diagnostic imaging, health care costs, and safety outcomes for patients with suspected appendicitis. In this parallel, cluster randomized trial, 17 community-based general emergency departments (EDs) in California, Minnesota, and Wisconsin were randomized to the AppyCDS intervention group or usual care (UC) group. Patients were aged 5 to 20 years, presenting for an ED visit with right-sided or diffuse abdominal pain lasting 5 days or less. We excluded pregnant patients, those with a prior appendectomy, those with selected comorbidities, and those with traumatic injuries. The trial was conducted from October 2016 to July 2019. AppyCDS prompted data entry at the point of care to estimate appendicitis risk using the pediatric appendicitis risk calculator (pARC). Based on pARC estimates, AppyCDS recommended next steps in care. Primary outcomes were CT, US, or any imaging (CT or US) during the index ED visit. Safety outcomes were perforations, negative appendectomies, and missed appendicitis. Costs were a secondary outcome. Ratio of ratios (RORs) for primary and safety outcomes and differences by group in cost were used to evaluate effectiveness of the clinical decision support tool. We enrolled 3161 patients at intervention EDs and 2779 patients at UC EDs. The mean age of patients was 11.9 (4.6) years and 2614 (44.0%) were boys or young men. RORs for CT (0.94; 95% CI, 0.75-1.19), US (0.98; 95% CI, 0.84-1.14), and any imaging (0.96; 95% CI, 0.86-1.07) did not differ by study group. In an exploratory analysis conducted in 1 health system, AppyCDS was associated with a reduction in any imaging (ROR, 0.82; 95% CI, 0.73- 0.93) for patients with pARC score of 15% or less and a reduction in CT (ROR, 0.58; 95% CI, 0.45-0.74) for patients with a pARC score of 16% to 50%. Perforations, negative appendectomies, and cases of missed appendicitis by study phase did not differ significantly by study group. Costs did not differ overall by study group. In this study, AppyCDS was not associated with overall reductions in diagnostic imaging; exploratory analysis revealed more appropriate use of imaging in patients with a low pARC score. ClinicalTrials.gov Identifier: NCT02633735.


Chronic unpredictable stress-induced reduction in the hippocampal brain-derived neurotrophic factor (BDNF) gene expression is antagonized by zinc treatment.
Preclinical data indicate the antidepressant activity of zinc and the involvement of the brain-derived neurotrophic factor (BDNF) in this mechanism. The present study investigates the effect of chronic (16 days) combined treatment with zinc (15 mg/kg zinc hydroaspartate) and imipramine (5 mg/kg) in chronic unpredictable stress (CUS) on the BDNF mRNA level in the rat brain. Moreover, serum zinc concentrations were also assessed. CUS induced a significant reduction in the BDNF mRNA level in the hippocampus by 21% but had no effect in the frontal cortex. Repeated treatment with zinc induced a significant increase in the BDNF mRNA level in the hippocampus in the unstressed animals by 12% and as in the chronically stressed animals by 14%, compared to the appropriate controls. Imipramine treatment did not affect this factor. However, combined treatment of zinc and imipramine induced a 12% elevation of the BDNF mRNA level in the stressed but not in the unstressed rats. CUS induced a 19% reduction in the serum zinc concentration, whereas combined treatment of zinc and imipramine reduced this concentration by 24% in the unstressed and increased it (by 20%) in the stressed animals. These results indicate that: 1) CUS induces a reduction in the BDNF gene expression with a concomitant diminution of serum zinc concentration and 2) the CUS-induced reduction in the BDNF gene expression is antagonized by chronic treatment with zinc.


Receptor subtype-dependent positive and negative modulation of GABA(A) receptor function by niflumic acid, a nonsteroidal anti-inflammatory drug.
In addition to blocking cyclooxygenases, members of the fenamate group of nonsteroidal anti-inflammatory drugs have been proposed to affect brain GABAA receptors. Using quantitative autoradiography with GABAA receptor-associated ionophore ligand [35S]t-butylbicyclophosphorothionate (TBPS) on rat brain sections, one of the fenamates, niflumate, at micromolar concentration was found to potentiate GABA actions in most brain areas, whereas being in the cerebellar granule cell layer an efficient antagonist similar to furosemide. With recombinant GABAA receptors expressed in Xenopus laevis oocytes, we found that niflumate potentiated 3 microM GABA responses up to 160% and shifted the GABA concentration-response curve to the left in alpha1beta2gamma2 receptors, the predominant GABAA receptor subtype in the brain. This effect needed the gamma2 subunit, because on alpha1beta2 receptors, niflumate exhibited solely an antagonistic effect at high concentrations. The potentiation was not abolished by the specific benzodiazepine site antagonist flumazenil. Niflumate acted as a potent antagonist of alpha6beta2 receptors (with or without gamma2 subunit) and of alphaXbeta2gamma2 receptors containing a chimeric alpha1 to alpha6 subunit, which suggests that niflumate antagonism is dependent on the same transmembrane domain 1- and 2-including fragment of the alpha6 subunit as furosemide antagonism. This antagonism was noncompetitive because the maximal GABA response, but not the potency, was reduced by niflumate. These data show receptor subtype-dependent positive and negative modulatory actions of niflumate on GABAA receptors at clinically relevant concentrations, and they suggest the existence of a novel positive modulatory site on alpha1beta2gamma2 receptors that is dependent on the gamma2 subunit but not associated with the benzodiazepine binding site.


The diagnostic value of immunohistochemistry in the typing of renal tumors with eosinophylic cytoplasma.
In this study, we aimed to review the diagnostic approach to eosinophilic cell renal neoplasms by light microscopy and immunohistochemical techniques. In this study 23 of these tumors were eosinophilic variant classic RCC, 15 eosinophilic variant papillary RCC, 13 eosinophilic variant chromophobe RCC and 13 oncocytoma cases. These tumors were immunohistochemically treated with CK7, CD117, EpCAM, Vimentin, RCCm (Renal cell carcinoma marker) and GST-α. In our study, contrary to the general literature on Vimentin, 65.2 % negativity was found in our patients with eosinophilic variant classic RCC. However, when compared with other tumor types in our study, vimentin expression was highest in eosinophilic variant classical RCC with 34.8 %. Statistically; RCCm, GST-α, EpCAM, CD117, CK7 were found to be significantly associated with tumor types, while no significant relationship was found between Vimentin and tumor types. RCCm positivity and CK7 and CD117 negativity were in favour of eosinophilic variant classical RCC, EpCAM, CK7 and CD117 positivity and Vimentin, GST-α and RCCm negativity supported eosinophilic variant chromophobe RCC, CK7 and RCCm positivity and CD117 and GST-α negativity were found in favour of eosinophilic variant papillary RCC. CD117 positivity and Vimentin, CK7 and GST-α negativity were found to support oncocytoma. The panel with RCCm, GST-α, EpCAM, CD117, CK7 will contribute to the differentiation of eosinophilic cytoplasm renal tumors that cannot be determined by morphological findings and to reach the correct diagnosis (Tab. 3, Fig. 4, Ref. 54). Text in PDF www.elis.sk Keywords: RCC, oncocytoma, immunohistochemistry.


Phorbol esters inhibit smooth muscle contractions through activation of Na(+)-K(+)-ATPase.
1. The role of protein kinase C (PKC) in agonist-induced contractions of guinea-pig ileum longitudinal smooth muscle has been investigated. 2. The phorbol esters, phorbol 12,13-dibutyrate (PDBu), phorbol 12,13-diacetate (PDA) and phorbol 12-myristate 13-acetate (PMA), relaxed tissues precontracted by submaximal concentrations of carbachol, histamine or substance P. 3. This inhibitory action of the phorbol esters was reversed following the application of ouabain, a specific inhibitor of Na(+)-K(+)-ATPase. Similarly, pretreatment with ouabain inhibited the ability of phorbol esters to relax tissues precontracted by the above agonists. 4. The slow relaxation of the tonic component of contraction induced by submaximal concentrations of carbachol and histamine, and all concentrations of substance P, was abolished in the presence of ouabain. 5. In Na(+)-loaded tissues, PDBu and carbachol caused a concentration-dependent increase of Na(+)-K(+)-ATPase activity, assessed by ouabain-sensitive 86Rb(+)-uptake. Extrusion of Na+, assessed by the cellular content of the ion, was also stimulated by PDBu (the effect of carbachol was not investigated). 6. We conclude that phorbol esters inhibit the tonic component of contractions induced by submaximal concentrations of these agonists through activation of Na(+)-K(+)-ATPase. We suggest that PKC may exert feedback control over the tonic component of agonist contractions through stimulation of the pump.


Effect of temperature and substitution on Cope rearrangement: a symmetry perspective.
Many reactions feature symmetry variation along the reaction path on the potential energy surface. The interconversion of the point group symmetry of the stationary points can be characteristic of these processes. Increasing the temperature, however, leads to the loss of symmetry in its traditional yes-no language. We find that in such cases the instantaneous distance of the molecular structure from its symmetric counterpart is a suitable collective variable that can describe the reaction process. We show that this quantity, the continuous symmetry measure (CSM), has a positive linear relationship with temperature, implying that even highly symmetric molecules should be considered as asymmetric above 0 K. Using ab initio molecular dynamics, we simulate the temperature-induced Cope rearrangements of several fluxional molecules and employ different CSMs to follow the reaction progress. We use this methodology to demonstrate the validity of important concepts governing these reactions: Woodward-Hoffmann rules and TS aromaticity. Statistical analysis of the CSM distributions reveals that ligands connected to the carbon frame have profound effect on the reaction course. In particular, our results show that lower temperatures tend to enhance the differences between the TS-stabilizing effect of the substituents.


The contribution of ion channels in input-output plasticity.
Persistent changes that occur in brain circuits are classically thought to be mediated by long-term modifications in synaptic efficacy. Yet, many studies have shown that voltage-gated ion channels located at the input and output side of the neurons are also the subject to persistent modifications. These channels are thus responsible for intrinsic plasticity that is expressed in many different neuronal types including glutamatergic principal neurons and GABAergic interneurons. As for synaptic plasticity, activation of synaptic glutamate receptors initiate persistent modification in neuronal excitability. We review here how synaptic input can be efficiently altered by activity-dependent modulation of ion channels that control EPSP amplification, spike threshold or resting membrane potential. We discuss the nature of the learning rules shared by intrinsic and synaptic plasticity, the mechanisms of ion channel regulation and the impact of intrinsic plasticity on induction of synaptic modifications.


FGF-1 induces ATP release from spinal astrocytes in culture and opens pannexin and connexin hemichannels.
Spinal astrocytes are coupled by connexin (Cx) gap junctions and express pannexin 1 (Px1) and purinergic receptors. Fibroblast growth factor 1 (FGF-1), which is released in spinal cord injury, activated spinal astrocytes in culture, induced secretion of ATP, and permeabilized them to relatively large fluorescent tracers [ethidium (Etd) and lucifer yellow (LY)] through "hemichannels" (HCs). HCs can be formed by connexins or pannexins; they can open to extracellular space or can form gap junction (GJ) channels, one HC from each cell. (Pannexins may not form gap junctions in mammalian tissues, but they do in invertebrates). HC types were differentiated pharmacologically and by Px1 knockdown with siRNA and by use of astrocytes from Cx43 knockout mice. Permeabilization was reduced by apyrase (APY), an ATPase, and by P2X(7) receptor antagonists, implicating secretion of ATP and autocrine and/or paracrine action. Increased permeability of cells exposed to FGF-1 or ATP for 2 h was mediated largely by Px1 HCs activated by P2X(7) receptors. After a 7-h treatment, the permeability was mediated by both Cx43 and Px1 HCs. FGF-1 also caused reduction in gap junctional communication. Botulinum neurotoxin A, a blocker of vesicular release, reduced permeabilization when given 30 min before FGF-1 application, but not when given 1 h after FGF-1. We infer that ATP is initially released from vesicles and then it mediates continued release by action on P2X(7) receptors and opening of HCs. These changes in HCs and gap junction channels may promote inflammation and deprive neurons of astrocyte-mediated protection in spinal cord trauma and neurodegenerative disease.


Are assortative mating and genital divergence driven by reinforcement?
The evolution of assortative mating is a key part of the speciation process. Stronger assortment, or greater divergence in mating traits, between species pairs with overlapping ranges is commonly observed, but possible causes of this pattern of reproductive character displacement are difficult to distinguish. We use a multidisciplinary approach to provide a rare example where it is possible to distinguish among hypotheses concerning the evolution of reproductive character displacement. We build on an earlier comparative analysis that illustrated a strong pattern of greater divergence in penis form between pairs of sister species with overlapping ranges than between allopatric sister-species pairs, in a large clade of marine gastropods (Littorinidae). We investigate both assortative mating and divergence in male genitalia in one of the sister-species pairs, discriminating among three contrasting processes each of which can generate a pattern of reproductive character displacement: reinforcement, reproductive interference and the Templeton effect. We demonstrate reproductive character displacement in assortative mating, but not in genital form between this pair of sister species and use demographic models to distinguish among the different processes. Our results support a model with no gene flow since secondary contact and thus favor reproductive interference as the cause of reproductive character displacement for mate choice, rather than reinforcement. High gene flow within species argues against the Templeton effect. Secondary contact appears to have had little impact on genital divergence.


Optimizing the mix of basic dental services for Southern Thai schoolchildren based on resource consumption, service needs and parental preference.
To identify the optimal levels and mix of basic dental services (sealants and fillings for permanent teeth and extraction of primary teeth) under two different dental settings: hospital-based and mobile dental clinics under specified resource constraints. A linear programming model is used based on explicit identification of system objectives and resource constraints. The objective was to maximize benefits as measured by parental willingness to pay (WTP) for basic dental services provided to schoolchildren subject to constraints on total resources, service need and parental preferences among different dental care settings. Optimization was identified to require 270, 180, 552, 828, 228 and 532 cases of hospital sealant, mobile sealant, hospital filling, mobile filling, hospital extraction and mobile extraction, respectively. The corresponding current service levels were 48, 281, 191, 170, 479, and 677 respectively. The optimal service configuration produced a total WTP of 485 860 baht which exceeded the WTP for the current service configuration by more than 75.4%. Mobile clinic fillings were the highest priority among basic dental services. The current service configurations fail to reflect the setting preferences and provide greater emphasis to extractions than the optimal configuration with less emphasis given to preventive and restorative services.


Adenylate energy charge in Saccharomyces cerevisiae during starvation.
Bakers' yeast cells, Saccharomyces cerevisiae, if grown aerobically on ethanol or if grown aerobically on glucose and allowed to pass into stationary phase, with utilization of accumulated ethanol, maintain a normal value (0.8 to 0.9) of the adenylate energy charge during prolonged starvation. In contrast, cells grown anaerobically on glucose and cells in the early stages of aerobic growth on glucose exhibit a rapid decrease of energy charge if transferred to medium lacking on energy source. These results suggest that functional mitochondria or enzymes of balance of adenine nucleotides during starvation. Yeast cells remain viable at energy charge values below 0.1, in marked contrast to results previously obtained with Escherichia coli. In other respects, the engery charge responses of yeast to starvation and refeeding are generally similar to those previously reported for E. coli.


Engineering dual-crystal configurations in perovskite oxides boosts electrocatalysis of lithium-oxygen batteries.
Sculpting crystal configurations can vastly affect the charge and orbital states of electrocatalysts, fundamentally determining the catalytic activity of lithium-oxygen (Li-O2) batteries. However, the crucial role of crystal configurations in determining the electronic states has usually been neglected and needs to be further examined. Herein, we introduce orthorhombic and trigonal system into 0.5La0.6Sr0.4MnO3-0.5LaMn0.6Co0.4O3 (LSMCO) by selectively incorporating Sr and Co cations into the LaMnO3 framework during the sol-gel process, which is used to explore the relationship among crystal structure, electronic states and catalytic performance. Based on both experimental and theoretical calculations, the dual-crystal configurations induce strong lattice distortion, which promotes MnO6 octahedra vibration and shortened MnO bonds. Furthermore, the suppressed Jahn-Teller distortion weakens the orbital arrangement and accelerates the charge delocalization, leading to the conversion of Mn3+ to Mn4+ and optimized electronic states. Ultimately, this resulted in optimized Mn 3d and O 2p orbital hybridization and activated lattice oxygen function, leading to a significant improvement in electrocatalytic activity. The LSMCO catalyzed Li-O2 battery achieves enhanced discharge capacity of 14498.7 mAh/g and cycling stability of 258 cycles. This work highlights the significance of inner structure and presents a feasible strategy for engineering crystal configurations to boost electrocatalysis of Li-O2 batteries.


Coupling microcirculation to systemic hemodynamics.
To discuss the role of microcirculatory abnormalities in critically ill patients and the link between systemic hemodynamics and microvascular perfusion. Microcirculatory alterations have been repeatedly observed in patients with severe sepsis, but recent findings show that these also occur in patients with severe heart failure and in those submitted to high-risk surgery. More severe and more persistent alterations are observed in patients with a poor outcome. Even though a minimal cardiac output and arterial pressure is mandatory to sustain the microcirculation, this level is not yet well defined and seems to be submitted to high individual variability. Above this level, microcirculation and systemic circulation are relatively dissociated, so that microcirculatory alterations can be observed even when systemic hemodynamics are within satisfactory goals. In addition, the response of the microcirculation to therapeutic interventions is often dissociated from systemic effects. However, microcirculatory perfusion can be affected by cardiac output and arterial pressure when these are critically altered. Microvascular alterations frequently occur in critically ill patients and these may be implicated in the development of organ failure and are associated with outcome. The link between systemic hemodynamics and microcirculation is relatively loose.


Stimulating whole system redesign: Lessons from an organizational analysis of the Whole System Demonstrator programme.
Whole system integration of health and social care has been positioned as key to improving care, increasing efficiency and controlling costs. However, evidence for the benefits of whole system integration is scarce. Drawing on organizational theory, this study uses the implementation of remote care services, viewed as an enabler for whole system working, to explore the reality of achieving this policy objective. Qualitative, longitudinal data were collected across nine UK sites adopting remote care over three years. Three sites formed the Department of Health's Whole Systems Demonstrator (WSD) programme for remote care. In addition, the implementation of remote care was explored in six other sites unconstrained by the randomized control trial procedures of the WSD programme. The methods were ethnographic (including 235 hours of observations and 184 interviews). Participants were health and social care staff and Government policy makers. Remote care did not lead to system redesign; however, local 'ownership' of new services did lead to more collaborative practices across the care system. Lack of integration was an enduring and endemic challenge across all sites, relating to differences in statutory responsibilities, absence of shared budgets and hybrid organizational roles, differences in work practices and organizational philosophies, and ambiguity around what 'whole system working' actually entailed. Policy initiatives like the WSD programme provide opportunities to phase in collaborative practices and create an awareness of the need for joint working. However, the progress observed suggests that the concept of whole system redesign around remote care is currently unrealistic.


Cell proliferation of myeloma plasma cells: comparison of the blood and marrow compartments.
The plasma cell labeling Index (PCLI), a measure of marrow plasma cells in S phase of the cell cycle, provides a good estimate of the proliferative capacity of the malignant clonal plasma cells. Multiple studies have also demonstrated its prognostic value in multiple myeloma (MM). Clonal plasma cells have been demonstrated in the peripheral blood of patients with multiple myeloma. The labeling index of these circulating cells can also be determined by a method similar to that employed for marrow plasma cells. The goal of this study was to compare the proliferation kinetics of plasma cells in the marrow to that in the circulation. The PCLI was estimated on peripheral blood (PBLI) and bone marrow aspirates (BMLI) using a slide-based bromodeoxyuridine (BrdU) immunofluorescence microscopy technique. One hundred seventeen patients who had simultaneous estimation of the PBLI and BMLI and who had > or =90% of the blood cIg+ cells identified as monoclonal plasma cells were studied. The PBLI was generally lower than the BMLI (median difference 0.4) and correlated with the BMLI (rho = 0.491, P < 0.0001). Twenty-seven patients with PBLI higher than BMLI had a poorer prognosis, with median survival of 2 months from the time of the labeling index determination, compared to 12 months for the remaining 90 patients (P = 0.01). Myeloma plasma cells circulating in the peripheral blood are capable of proliferation, albeit at a slower rate compared to those in the marrow. These differences reflect a more favorable microenvironment in the marrow likely resulting from the differences in the cytokine levels and supporting stromal cells in the marrow.


The effect of impaired sleep on preterm labour.
Sleep disturbance has become an important health problem for pregnant women. In fact, pregnancy-associated sleep disorder has been recognized as a distinct clinical entity. We aimed to study the relationship between sleep disturbance and preterm birth during pregnancy in a sample of Iranian women. In this analytical cohort study, 231 pregnant women in their 28th-32nd gestational week were recruited, using the multistage sampling method, from four healthcare centres in Ardabil, Iran, during 2010. The women were followed-up until 37-week gestation. One hundred and twelve women did not have sleep disturbances while 119 women had sleep disturbances. The Insomnia Severity Index (ISI), Epworth Sleepiness Scale (ESS), and a demographic data questionnaire were used for data collection. Data were analysed using SPSS software. Descriptive statistics, t, Chi-square, Fisher's exact, and Mann-Whitney tests were used as appropriate. The prevalence of preterm labour was 11.8% in women with sleep disorder compared with 11.6% in women without sleep disorder (p = 0.9). Sleep duration less than eight hours, daytime dysfunction and impaired quality of life as a component of ISI showed a significant relationship with preterm birth (p = 0.02, p = 0.044, and p = 0.047, respectively). Although daily dysfunction and lower quality of life because of sleep problems, and total sleep duration were variables associated with preterm birth, we found no significant relationship between sleep disorder and preterm birth.


Demosaicing using optimal recovery.
Color images in single-chip digital cameras are obtained by interpolating mosaiced color samples. These samples are encoded in a single-chip charge-coupled device by sampling the light after it passes through a color filter array (CFA) that contains different color filters (i.e., red, green, and blue) placed in some pattern. The resulting sparsely sampled images of the three-color planes are interpolated to obtain the complete color image. Interpolation usually introduces color artifacts due to the phase-shifted, aliased signals introduced by the sparse sampling of the CFAs. This paper introduces a nonlinear interpolation scheme based on edge information that produces high-quality visual results. The new method is especially good at reconstructing the image around edges, a place where the visual human system is most sensitive.


Lysinibacillus Isolate MK212927: A Natural Producer of Allylamine Antifungal 'Terbinafine'.
Resistance to antifungal agents represents a major clinical challenge, leading to high morbidity and mortality rates, especially in immunocompromised patients. In this study, we screened soil bacterial isolates for the capability of producing metabolites with antifungal activities via the cross-streak and agar cup-plate methods. One isolate, coded S6, showed observable antifungal activity against Candida (C.) albicans ATCC 10231 and Aspergillus (A.) niger clinical isolate. This strain was identified using a combined approach of phenotypic and molecular techniques as Lysinibacillus sp. MK212927. The purified metabolite displayed fungicidal activity, reserved its activity in a relatively wide range of temperatures (up to 60 °C) and pH values (6-7.8) and was stable in the presence of various enzymes and detergents. As compared to fluconazole, miconazole and Lamisil, the minimum inhibitory concentration of the metabolite that showed 90% inhibition of the growth (MIC90) was equivalent to that of Lamisil, half of miconazole and one fourth of fluconazole. Using different spectroscopic techniques such as FTIR, UV spectroscopy, 1D NMR and 2D NMR techniques, the purified metabolite was identified as terbinafine, an allylamine antifungal agent. It is deemed necessary to note that this is the first report of terbinafine production by Lysinibacillus sp. MK212927, a fast-growing microbial source, with relatively high yield and that is subject to potential optimization for industrial production capabilities.


Classical toy models for the monopole shift and the quadrupole shift.
The penetration of s- and p(1/2)-electrons into the atomic nucleus leads to a variety of observable effects. The presence of s-electrons inside the nucleus gives rise to the isotope shift in atomic spectroscopy, and to the isomer shift in Mössbauer spectroscopy. Both well-known phenomena are manifestations of the more general monopole shift. In a recent paper (Koch et al., Phys. Rev. A, 2010, 81, 032507), we discussed the existence of the formally analogous quadrupole shift: a tensor correction to the electric quadrupole interaction due to the penetration of relativistic p(1/2)-electrons into the nucleus. The quadrupole shift is predicted to be observable by high-accuracy molecular spectroscopy on a set of 4 molecules (the quadrupole anomaly). The simple physics behind all these related phenomena is easily obscured by an elaborate mathematical formalism that is required for their derivation: a multipole expansion in combination with perturbation theory, invoking quantum physics and ideally relativity. In the present paper, we take a totally different approach. We consider three classical 'toy models' that can be solved by elementary calculus, and that nevertheless contain all essential physics of the monopole and quadrupole shifts. We hope that this intuitive (yet exact) analysis will increase the understanding about multipole shift phenomena in a broader community.


[Femoral neck morphology differentiates femoral neck from vertebral or hip osteoporotic fracture].
Mineral density and bone geometry are the two main biomechanical factors related to bone resistance to stress. In this study we assess whether differences in the proximal femur geometry (PFG) characterize different types of osteoporotic fractures. We studied 533 postmenopausal women aged 50 - 85. They were divided into four groups matched for bone mineral density at the femoral neck; without fractures (165), vertebral fractures (139), trochanteric fractures (102), and hip fractures (127). Dual X-Ray absorptiometry (DXA) scans at the spine and at the hip were carried out to measure bone mineral density and the DXA images were used to define the proximal femur geometry parameters of the hip. Age, height, vertebral BMD and PFG parameters (i.e. femoral neck-shaft angle (NSA) and hip axis length (HAL) were different when all four groups were compared by the Anova test. Patients with vertebral fractures were then compared by multivariate analysis to those with trochanteric fractures. The variables that discriminated the two groups were: age, age at menopause, weight, height, and vertebral BMD, but not PFG. Comparing vertebral to hip fractures the distinguishing variables were: vertebral BMD, height, NSA and HAL. We found that hip fractures had longer HAL and wider NSA than vertebral fractures, whereas no statistically significant differences were found between trochanteric fractures and vertebral fractures concerning PFG. These data indicate that differences in PFG parameters might have a role in predisposing to femoral neck fracture.


The overview of development of novel bacterial topoisomerase inhibitors effective against multidrug-resistant bacteria in an academic environment: From early hits to in vivo active antibacterials.
Antimicrobial resistance caused by the excessive and inappropriate use of antibacterial drugs is a global health concern. Currently, we are walking a fine line between the fact that most bacterial infections can still be cured with the antibiotics known so far, and the emergence of infections with bacteria resistant to several drugs at the same time, against which we no longer have an effective drug. Therefore, new antibacterial drugs are urgently needed to curb the hard-to-treat infections. Our group has developed new antibacterials from the class of novel bacterial topoisomerase inhibitors (NBTIs) that exhibit broad-spectrum antibacterial activity. This article reviews our efforts in developing highly potent NBTIs over the past decade. Following the discovery of an initial hit with potent enzyme inhibitory and broad-spectrum antibacterial activity, an extensive hit-to-lead campaign was conducted with the goal of optimizing physicochemical properties, reducing hERG inhibition, and maintaining antibacterial activity against both Gram-positive and Gram-negative bacteria, with a focus on methicillin-resistant Staphylococcus aureus (MRSA). This optimization strategy resulted in an amide-containing, focused NBTI library with compounds exhibiting potent antibacterial activity against Gram-positive bacteria, reduced hERG inhibition, no cardiotoxicity in in vivo zebrafish model, and favorable in vivo efficacy in a neutropenic murine thigh infection model for MRSA infections.


A Bayesian predictive sample size selection design for single-arm exploratory clinical trials.
The aim of an exploratory clinical trial is to determine whether a new intervention is promising for further testing in confirmatory clinical trials. Most exploratory clinical trials are designed as single-arm trials using a binary outcome with or without interim monitoring for early stopping. In this context, we propose a Bayesian adaptive design denoted as predictive sample size selection design (PSSD). The design allows for sample size selection following any planned interim analyses for early stopping of a trial, together with sample size determination before starting the trial. In the PSSD, we determine the sample size using the method proposed by Sambucini (Statistics in Medicine 2008; 27:1199-1224), which adopts a predictive probability criterion with two kinds of prior distributions, that is, an 'analysis prior' used to compute posterior probabilities and a 'design prior' used to obtain prior predictive distributions. In the sample size determination of the PSSD, we provide two sample sizes, that is, N and N(max) , using two types of design priors. At each interim analysis, we calculate the predictive probabilities of achieving a successful result at the end of the trial using the analysis prior in order to stop the trial in case of low or high efficacy (Lee et al., Clinical Trials 2008; 5:93-106), and we select an optimal sample size, that is, either N or N(max) as needed, on the basis of the predictive probabilities. We investigate the operating characteristics through simulation studies, and the PSSD retrospectively applies to a lung cancer clinical trial. (243)


Autism susceptibility candidate 2 (Auts2) encodes a nuclear protein expressed in developing brain regions implicated in autism neuropathology.
Autism susceptibility candidate 2 (Auts2) is a gene associated with autism and mental retardation, whose function is unknown. Expression of Auts2 mRNA and protein were studied in the developing mouse brain by in situ hybridization, immunohistochemistry, and western blotting. Auts2 mRNA was highly expressed in the developing cerebral cortex and cerebellum, regions often affected by neuropathological changes in autism, and a few other brain regions. On embryonic day (E) 12, Auts2 mRNA was expressed in the cortical preplate, where it colocalized with Tbr1, a transcription factor specific for postmitotic projection neurons. From E16 to postnatal day 21, Auts2 was expressed most abundantly in frontal cortex, hippocampus and cerebellum, including Purkinje cells and deep nuclei. High levels of Auts2 were also detected in developing dorsal thalamus, olfactory bulb, inferior colliculus and substantia nigra. Auts2 protein showed similar regional expression patterns as the mRNA. At the cellular level, Auts2 protein was localized in the nuclei of neurons and some neuronal progenitors.


Regulation of NF-kappa B activation in T helper 1 and T helper 2 cells.
In most cell types, NF-kappa B is activated by release from a cytoplasmic inhibitor protein, I kappa B, followed by its translocation to the nucleus where it binds to the regulatory regions of many genes, including the IL-2 gene in T lymphocytes. We have previously shown by electrophoretic mobility shift assays that nuclear extracts prepared from activated, non-IL-2-producing Th2 cell clones. We show here that Th-1 and Th2 cells have similar levels of cytoplasmic p65(RelA) and p50, but TCR stimulation fails to induce the nuclear translocation of p65(RelA) in Th2 cells. Nuclear translocation of p65(RelA) can be induced by IL-1 stimulation of Th2 cells, indicating that a basic mechanism of NF-Kappa B activation common to many cells is intact in Th2 cells. We demonstrate that IL-1 and TNF induce rapid nuclear translocation of p65(RelA) in T cell clones, whereas TCR-induced NF-Kappa B activation in Th1 cells is delayed and may be longer in duration. This suggests that the TCR pathway of NF-Kappa B activation is different from the cytokine pathway. Furthermore, we show that Th1 and Th2 cells express different levels and/or different forms of I kappa B alpha, and that cytokines, but not TCR stimuli, significantly modulate detectable levels of cytoplasmic I kappa B alpha.


Immune modulation of goat monocytes by Fasciola gigantica Legumain-1 protein (Fg-LGMN-1).
Legumains belonging to C_13 peptidase family of proteins, and are ubiquitously disseminated among all vertebrate and invertebrate organisms, and have been implicated in innumerable biological and cellular functionality. Herein, we characterized and evaluated immunoregulatory characteristics of Legumain-1 from Fasciola gigantica (Fg-LGMN-1) during its interaction with host immune cells. The isopropyl-ß-d-thiogalactopyranoside (IPTG) stimulated RFg-LGMN-1 protein was positively detected by rat serum containing anti-RFg-LGMN-1 polyclonal antibodies. Furthermore, the uptake of RFg-LGMN-1 by goat monocytes was successfully confirmed using Immunofluorescence Assay (IFA). The immunohistochemical analysis revealed the native localization of LGMN-1 protein on the periphery and internal structures such as suckers, pharynx, and genital pore of the adult parasite, thereby validating its presence in excretory-secretory (ES) products of F. gigantica. The RFg-LGMN-1 co-incubated with concanavalin-A (Con-A) stimulated the increase of interleukin 2 (IL-2), IL-10, and IL-17 in monocytes derived from peripheral blood mononuclear cells (PBMCs) in the concentration-dependent manner. However, the IL-4 cytokine in response to the RFg-LGMN-1 protein declined. These results illuminated the role of LGMN-1 during the parasite-host interface. Our findings elaborated additional evidence that Legumain protein play a role in the manipulating host immune responses during parasite infections. However, further evaluation of RFg-LGMN-1 protein in context of its immunomodulatory roles should be conducted to enhance our understandings of the mechanisms employed by F. gigantica to evade host immune responses.